Belém VIRUS Laboratory

Belém
VIRUS
Laboratory
Belém, Pará, Brazil
1965 A N N U A L R E P O R T

Belém
VIRUS
Laboratory
Belém, Pará, Brazil
1965 A N N U A L R E P O R T

This report is reproduced as received
from the director af the labaratary

A.QUE OVIRUS~ l:rrINE
[
11 14 1.5 16 17 19 23 24 26
CONTENTS .-
INTRODUCTION .o o
YELLOW FEVER
vmus SURVEIW.NCE o o .
BIRD RECAPrURE PROGRA1-1ME .o o o o .o
BATS
FIEID STUDIES IN AMAPÁ TERRITORY o 8 8
SERRA DO NAVIO
Results o
Neutralization testing in mice wi th the Tacaribe
Group agente o o .o o o ..
roRTO PLATCtI & MONGUBAS REGION
KAYAPO-GOROTIRE llmIAN SERUM SURVEY.
TISSTJE CULTURE STUDIFS ON ARBOVmUSES. o .o
SIMULTANEOUS ISOIATION Di MICE & TC o o
IIWLumCE OF TYPE OF AGAR ovrnIAY ON PIAQUE FORMA TI ON ...
bo~WARISON OF CHICK & TURKEY EMBRYO CULTmmS. o o .
rn GMK & HEp2 CEUS o o o o o
COMPLEMENT FIXATION & TISSUE CULTURE. o o
Detection or Amapar! (AN 70563) antigen o ..
2. Use oí combined antigen pools o
3. Tests with material fraro Labrea
NEUTRALIZATION TESTS DI TISSUE CULTURE
Oropouche
Group A .
Group B ..o o o
Phlebotomus group ..o
Cotia group o o n .o ..o
INHIBITION TESTS .to o o o ..~ .
~
1
4
6
8
13
13
20
21
22
22
22
23
24
25
25
26
26

49
ENTOMOLOGY
HumaJl bait catches ..o
The ~.n Blower Trap
Comparison or baits
Mbuse bait -hood catches
Mbuse bait -blower trap catcheso
Mbuse bait -Trinidad NQ 10 trap. o
Chicken bait -}ngoon trap.
Light trap catches o o
Suction trap catches
Phlebotomus o ...o ..o
Precipitin tests on blood meals.
Ticks o o o o
PARASITOLOOY o o o o
MDUSE NEUTRALIZATION TESTING o o ...
Group B ...o .o .o o ..o o
Changuinola group ..o o.
AN 84381 (unidentified
IABOI~TORY CONTAJ:IINATION CHECK .o
REPOR'l'ED EQUDJE ENCEPHALITIS m BRAGANÇA
CATTLE DISEASE o .
1!()I-nmYS o o
1-1QUSE COLONY o
l-brtali ty o
Attempts at increasing output
ReIM.rks o ..o o
-iv-
27
28
29
29
32
33
36
37
37
38
38
39
39
40
40
40
42
43
44
44
45
46
47
47
SI

{
HISTOPATHOLOGY 51
Histopathology or wiId animaIs 51
Histopathology of sentinel mice 52
Experimental pathology in mice 52
Other work
BEL:E1IJ1: VIRUS LABORATORY STAFF, 1965 54
VISITORS . 56
TABLFS ..
FIGURES
APPENDIX I. THE SEROLOGICAL RESPONSE OF ANn.fALS
TO VIRUS INFECTION IN lrrINGA FOREST 131
APPENDIX II. REPORT ON THE LA.BREA REGIOM EPID~C,
SEPT:F)ABER -NOvmvffiER 1965
APPENDIX li! VIRUS RESEARCH IN AMAZONIA
v
52
58

vector,
DITRODUCTION
With the departure of Dr. Robert E. Shope from this laboratory in early June
year.. a link with the pioneering days oí the Causeys was brolcen, but the local
staff carried on the routine dependably, most of thern having been with the laboratory
from the beginning.
Tables 1, 2 and 3 show the current status of sources and numbers of isolations of
individual viruses to date. Natable additians this year are the first isalatians af a
new Simbu group virus (utinga virus from a sloth, of the Belem type of Bushbush virus
mos qui toes (~~ spp. , and af Apeu virus fraro a wild animal (Calluram;ys ).
Rainfall has been average (Table 9) except for the months of r"áy and October which were
well above, and Ju~, August and November which were below average; total rainfall was
average, and 1ess than 1964. In spi te or the continued heavy rain in May the number or
viruses isolated declined, and in July there were no vertebrate or arthropod isolations
made from the Belem area, and only half-a-dozen sentinel mouse isolates. The bird
recapture programme at the IPFAN forest in July and August also failed to turn up more
than one virus (MUcambo) and a few not very convincing serological conversions, in
contrast to last year, and a large bat netting operation produced no virus isolations
and ve~ few serological positivcs.
Following the isolation of yello~ rever virus from the sentinel monkey in Utinga
in December 1964~ intensive studies were made to try to incriminate host and
the on1y pasitive finding was af haemagglutinatian inhibiting ant1bodies in
10/30 monkeys.

1961. Drs.
-2-
The Amapá field progr8Dme prodUCOO strains of AN 70563 (Amapari) virus from
Neac~ and Oryz~ regular~, mn a strain was also i8o1ated from gamasid mites
combed from these rats, but we ~ without a sati8factoI")" serological system for th1s
virus, which ~ an mSO well above its low WSO 1n our mice. A collection of
,
Haemgo~ spp. f'rom Amapa 7ielded Tacaiuma virus, not seen 1n this laboratory- since
A serious decline 1n the productiviV of the mouse colOr1Y' delay'ed isolation and
identification 'Work 1n the secom ha1f of the year. The mice were fow1d to be
infected with salmnellae and a number or other enterlc agents and a180 heavi1,)"
1nfested with a cestode (~olepis ap.). Measures were taken to provide .fil tered
drinking water for 'Ule breeding colonies and to destroy the insect alterna tive hosts
(,Forficularidae) ot the cesto.es.
A new wooden laboratory- has been built in the IPEAN forest to 8Upersede the old
thatch cabin used by the ento1lK)1ogists am orni tholog1sts. It has a raised fioor
6 x 4 me, a walled-oíí section for atores, laboratory bmlchea, hammock hookB, car
battery- light1ng, atn a ra1nwater 8Upply from an outaide tank. It 18 comp1ete~
screened o A new msquito trap on the blower princip1e, Wich separates the catch
into 6-haur segments, has gone into operation.
Bragança.
In March reporta were received ar a new autbreak ar equine encephalitis at
Robert Shope, Francisco Pinheiro and Wilbur Downs (the last-named on
a visit to Belem) went to the area but saw no cases.
In the same mnth Drs .Francisco
Pinheiro, Wilbur Downs, Gilberta Bensabat.h and Jack Woodall, togetJ1er witJ1 Amelia
,
Andrade am Anm.zonia Toda, visited the field operation 1n Amapa Territo17.
,
In Ju1y we were asked by" the Serviço de Defesa Sani tãria Ani!Dal to help,
investigate siclmess and death in cattle following routine rabies vaccination.

-3 -
Several strains oí íoot-and-rnouth disease virus were isolated, but none oí rabies.
Arbovirus antibodies were rare.
In October, Drs. Jorge Boshell and Francisco Pinheiro visited Labrea at the
request aí the public health autharities in Manaus, to investigate a renewed
outbreak of the disease which has caused high infant mortali ty for many years.
A mother with her sick children were brought to Belem and studied intensively in
In December, Drs. Jorge Boshell and Uack \voodall were invited by the Força
Aérea Brasileira to visit the Kayapo-Gorotire Indians, and made a serological survey.
Amelia Andrade" head of the serolo~ section" spent 8 weeks in lbadan" Nigeria"
helping to organize the serology section at the new Arbovirus Research Unit there
which ia directed by Dr. ottis Causey. Amazonia Toda spent the secand halí aí the
year in são Paulo studying the taxono~ of ~~ 1vith Prof. o. Forattini. Dr.Francisco
Pllllleiro joinod an airborne Bolivian-U.S.forces medi cal team in April for a survey
or Brazilian border areas ror signs or Bolivian haemorrhagic rever, and Drs. Jack
Woodall and üorge Boshell visited the same team in the field in Mato Grosso in
Dr. Fhilip Humphrey returned to Belem for the third successive year to
operate a bird programme, and Dr. Charles Handley made a large collection or bata
during a week in August, from which we obtained much virological material.
During the year, training has been given to animal technicians, an entornologist,
and a veterinarian from other institutions.
The Belem Virus Laboratory is rnaintained jointly by the Fundação Serviço Especial
.
de Saúde PÚblica and The Rockefeller Foundation. Gratitude and appreciation is
extended to the Institu~~~~o Cruz for their part in the cooperative mammal field
prograJm11es, to the .!E~ mining com~ for their collabora tion in the field studies

encountered.
,
-4-
, ,
in Amapa Territory, to Dr. Jose f.~ria Conduru, director oí the _InBti~~Bquisas
e Experimentação Agropecuárias do Norte (IPEAN) for permission to work 11'1 the IPEAN
forest and to the Força Aérea Brasileira for transport and collaboration 11'1 the
investigations at Labrea and Itagorotirco
YE LLOW FEVER
On December 4th, 1964, ye11ow rever virus was recovered from the blood ai a
sentinel ~~ monkey in Utinga forest, and short~ after, two monkey skeletons
were found in the saroe study area where the mamma1 recapture prograro is in progress.
After Christmas and continuing through January, a more intensive study of the animal
and mos qui to fauna of utinga was undertaken. Cano~ daytime mosquito captures were
instituted resu1ting in co11ection of 1,016 mosquitoes, 613 of which were ~aemago~~o
Ten hunters were hired to shoot monkeys, squirrels, and any other arboreal animaIs
Birds and bats were mist-netted, and lizards and toads hand-caught at
the forest edge. In addi tion, the mamma1 recapture program continued to yield rodents
and marsupiaIs. In alI" 1,286 vertebrates were samp1ed in or near Utinga forest
during th.e 2 month period.
Mast ar these were toads (591
1izards (161) and
rodents (311 There were no recoveries of yellow rever virus from feral sources.
In addition, 1 sentinel chickens were exposed during the period without serological
conversion by HI to group B viruses.
HI study of the monkey sera revealed that 8 of 21 Tamarin and 2 of 9 Saimiri
had group B antibody compatible with previous yellow rever infectian. N tosting ic
in baby mice with yellow rever, Ilheus, Bussuquara and SLE viruses was done with 24
af the mankey sera, inc1uding 9 HI positives and 15 HI negativese All 9 positives
had LNIts aí 2.1 ar greater with yellaw rever and 2 aí these alsa neutralized

R!oech~ antibody. nheus, 'UF
5
Ilheus virus o There were no other neutralization indices of over 200 for the 4 group
B viroses tested.
No group B HI antibody was encountered in 57 bats, 1 porcupine, 1 ~~,
29 marsupiaIs (incIuding tree-captured animaIs), 4 squirreIs, 5 slaths, ar 1 turtIe.
T-wo of 36 birds ,-ri th group B antibody had higher ti ters to SLE than to yellow rever o
as in the past continue to have group B RI antibody, presumably because
af previaus Bussuquara infectian. One Qryzomys and 2 ~_~cto!r\Y:8 a180 had group B. HI
In no case was a high-titered, clear-cut yellow rever HI pattern observed
in the rodents or birds.
Twen~-two people living near or working in utinga forest were bled during
Januzry before vaccina tion was ini tia ted. Their group B serological reactions can be
divided into four categories L) sera neutralizing yellow rever virus, 2 which had
no group B HI antibody, anothe~ with group B HI antibody and which also neutralized
Ilheus virus, and a fourth wi th yellow rever RI antibody in the 1 :20 dilution and
having a LNI of 1.5 in ye11ow rever N testing, 2) sera neutralizing Ilheus virus,
5 which had a primary infedtion HI pattern with Ilheus, 2 which had a broadly
reacting HI pattern, and one with a broadly reacting HI pattern and L~I af 1.6 with
3) a serum neutralizing SLE virus with a broad~ reacting group B RI pattem,
anã 4) 9 sera which were negative by HI and N test wi th yellow rever, Ilheus, SLE,
and Bussuquara, and an additional serum negative ~ HI and not tested by N. For
yello1i rever virus there were no HI ti ters over 1 :20 and the 2 sera which neutralized
only yellow fever viros among the group B agentA'3 had INI of on1y 1.9. There ia thus
little evidence pointing to recent yelloti rever infections in the human population
of the Utinga forest region and a search for clinical yellow rever in Janua~ was
unfrui tful.
testing was dane with the 3 human sera positive in N testo One was anti-
complementary and the other two were nega tive in the 1:8 dilution. CF testing of

centineIa, 15%
6
It is c1ear that ye11ow rever virus 'fas active in utinga forest during 1964.
The only positive serological evidence for recent vertebrate infection was in the
for
mop~ey population. The epizootic would have gane undetected if it were not jthe
routine surveillance studies in progresso
This work has been vublished by Bensabath ~. (1966) BoI. Oficll1a Sanit.
Panamer. 60:187-192-- "Recuperacion de virus amarilico, procedente de um mono
en Ias cercanas de BeIem, Brasil".
ROUT~ VIRUS SURVEIIJANCE
Tables 4 through 8 list the viruses isolated during the year, principally from
material collected during the routine surveillance activities in the IPCAN and utinga
There
were 68 virus is ala tions !roro IPEAN, 61 from utinga.
Guajará, Itaporanga and Aurá viruses were found at IPEAN but not at utinga, whilst
a \~eomyia complex virus was isolated at Utinga on~.
Rain gauges opera ted in both
areas from October, revealing that there may be a difference of as rnuch as 33% in the
monthly rainfall of these are as (Table 9)0
According to the sentL~el mouse isolations (Table 4), Caraparú vírus transmission
ceased between mid-June and mid-December; this is borne out by the lack or mosquito
and wild rodent isolations or this virus dl~ing the same period, aI though it was
recovered from the sentinel monkey in utinga in mid-october. This is quite different
!'ram 1961~, when sentinel mice yielded Caraparú virus every manth except the lOvl-"rlruS
montI'1S af I"b-y and June. In 1965,
campared with abaut 1% af the wild vertebrates pracessed.
of the sentinel mouse groups were infected,

y I.
enezuelensis
er ever,
was
7
A summa~ of 3 years' resul ts from the utinga mammal recapture programme is
given as Appendix
An interesting range record was found this year when a
1~sa murina murina (M 14799), first captured on the trap grid in February,
was recovered in August in a trap line set 2 km. distant, for the purpose of
providing some museum specimens o
Whilst -
rodents frequently urinate, and when one ~~ at
anaesthesia prior to heart puncture,
utinga rIas noted to have
haemorrhagic urine, this was collected and immediately inoculated into adult mice at
the Utinga laboratory. I-fucambo virus was isolated from this specimen, but not from
a blood specimen taken the following day. However, the rat developed MUcambo RI
antibodieso
Herpes virus has been recovered regularly in mice froro the oral aphthae oi
patients, and rabies was isolated from the brain of a dog (CA44) which died after
biting an Air Force officer; the latter was referred for vaccination.
Tab1e 8 shows that only 16 out of the more than 60 mosquito species co11ected
during the year were positive for virus. The genus ~~ produced both the
cornmonest and rarest virus -producing species:
with 2 isolates from
over 16,000 specimens, and M.titillans with an isolate !roro only 226 individuaIs.
the commonest species was not the best for vírus; Culex B 9 gave 12 iso1ations
aí 6 different viruses íram 12,000 specimens, and the mixture of unidentifiable
Culex spp.
.. gave 13 iso1ations oí 7 viruses írom about the same numbero Culex B 1
productive, with 6 iso1ates of 3 types from on1y 571 mosquitoes. Thus crude
infec ti on rates ranged from 1:100 to 1:8,000 mosquitoeso
Tab1e 13 shaws the campasitian af aver 10,000 ectaparasites inaculated during
the year, without resulto

.
u.s.
years.
-8 -
BnlD RECAPrURE ffiOORAMME
Dre Philip S. Humphrey returned to Belem this year with 3 assistants from the
After several days aí preliminary field wark (clearing trails, putting up nets,
etc.) the recapture program was begun at Station A (IPEAN study area
on June 30,
using approximately the same trail system and arrangement of mist nets as in previous
Twenty-seven days of field work were accomplished at Station A beginning
30 June and ending 1 Sept.
at about 6:00 p.m.
The normal day in the field started at 5:45 aom. and ended
The following pre1iminary summa~ does not inc1ude mention of 1) a sma11 number
of birds preserved as specimens for identification, 2) a small number of birds
which died in tl1e neta on June 30 and July 1 and one or two other days.
Ao ~gingand recaptures -During 30 June -1 Sept. 773 birds aí 105 species
were ringed ar narked o
one or more tiJneso
In addi tion 38 birds ringed June -Sept. 1964 were recaptured
(Discaunting marta1ity, 102 birds were ringed in 1964; 37.25% aí
these were recaptured in 1965. Recaptures ar ringed birds (ringed in 1964 and ringed
ar narked in 1965) were as follows in 1965:
Individuals recaptured
4 tiMes or more
3 times
2 times
1 time
Total indi viduals recaptured
13
28
54
142
one or more times. 237 ( .29g2% aí total
number aí b1rds ringed
ar marked jJ1 1965,
includiJ1~ 1964
recaptures).
Far~-ane ringed birds (5.1% aí total number af birds ringed
or marked in 1965, p1us 1964 recaptures) died accidenta1~ in nets, bird bags, or as a
result af bleeding.

wever, o
follows: 114
"9-
B. ~ghnets -Faur high neta were rigged at heights aí íram 17 to 24 metersj
installations were not completed until the latter part of JulYo Of the lO5 species
ar birds caught and ringed 30 JUne -1 Septo 33 species were caught from time to
tiIoo in the high nets and 14 species were caught on1y in the high nets.
af the 811
marked or ringed birds captured 30 June" 1 Sept., 124 were caught one or more times
in the high netso
00 Arbovirus studies -
~ ~~-
The total number of bloods collected (not alI were tested
ringed birds VIas 563. Inoculations were made of plasmas from bloods collected
during the momings into TC and baby mice. MUc amb o vírus was ísolated ín micc, but
not in TC, from a ~~ e~hrocephala. About 450 individual birds were bled as
N Q individuaIs
Total indi vid11als
Total individuaIs bled
335
82
25
7
.~
two or more times .
or the 114 individuaIs bled two or more times,
in titer of HI antibodies (Table 11
Q times bled
once
t"iice
three timesfour
times
showed conversions or changes
It is rather disturbing to find titres of up to 1:40 for EEE, for examp1e,
disappearing in a rnatter of weeks, and the titre of ltaporanga fluctuating 4-fold
between rebleedings. This might suggest that the apparent conversion to EEE in
Table 11 is only a fluctuation in pre-cxisting antibody. Even wi th SLE we have one
case oí a titre oí 1:20 disappearing and then reappearing as 1:40. But the Phlegopsis
conversion to ~ 1:320 for SLE appears genuine enough.
this gives liS clear
antibody evidence af the activity anly af SLE in birds during the periad af the study
and SLE virus was not isolated.

,
P.lthough Bussuquara.
responses:
-10
The 27 seroposi tives to ltaporanga (Tab1e 12) are interesting in that 17 oí
them are known to be cano~-haunt1ng birds, and 13 of those be1ong to the 2 genera
Rates for these genera are not yet calculated but could
run as high as the 36% oí positives found last year in Callur~~, an arboreal
mrsupialo Also there were 8 seropositives among the bats taken in August, tending
to confirm that transmission is usual~ arboreal.
a strain oí Mucambo was isolated, MUcambo antibody was scarce and
low-titred and general~ in association with other Group A antibody among the birds,
with the, exception of a ~~! which titred 1:160,
In total, 99/968 bird sera tested at 1:20 against 8 units oí 18 antigens gave
one or more positive resulta, which were against alI the antigens used except for
The 3 yellow rever positives were all positive to SLE in higher titre.
The great majority of titres were in the range 1:20-1:40 on1y. A study of
,
the Urubu,
Coragyp~, yie1ded 17 seropositives, with the fo11owing numbers of probab1y specific
EEE 2, MUcambo 1, SLE/Ilheus 10, Turlock 1, Itaporanga 2 and Jurona 5.
The high proportion of Group B positives confirma last yearls finding in ~~.
One plasma which was HI positive for 11Ucambo was confirmed by NTo A senlnl
virus stack aí ltaparapga (AN 64582 titring 5.3 log 1050 was obtained from mice bled
48 hrs. after inoculation, and used to test 39 plasmas, with the following results:
RI
1:201:10
O
NT results
Positiv"e Inconclusive Negative
9
6
1
Serum vírus stocks were also prepared in infant mice for Jurona (AR 40578
titring 5.3 (blee 24 hr. after inoculation) and 6.2 (48 hr.). These werc used to
test 47 k~ plasmas at final di1utions of 1:8 (not inactivated), of which 31 had been
bird
3
2
O
1
;;
12

EAN
cessory -48 th25th
RI posi tive for Juronao
-11 -
The resulta were all negative, but this is not conclusive
evidence that the HI positives were non-specific, since factors such as absence of
factor and lability of the virus preparation need to be considered.
A serum virus stock of Turlock (AN 32260) was obtained by bleeding 3-day-old
hours after inoculationo
The titres af these preparatians were 4.5 lag
ID50 (30 hrs.) and 3.8 log ID50 (48 hrs.). This 5 tock was used to test 27 bitd sera
(1964 co11ection) oí which 16 had been positive to this strain by HI test (7 oí
these on1y had titres or 1:10). The results were as follows:
NT
Negative
Inconclusive
Positive
BATS
~-
9
2
O
HI
1:10
2
O
.5
titre
1:20 ar greateE
Dr. Charles Handley oí the U.S.National MUseumvis i ted Belem frO~l
August and carried out 10 catches with rnist neta, with the following results:
(Station A) August 17th
104
18th
19th
22nd
74
51
54
ro
o
3 6
15-28
Utinga 20th
21st
26th
157
150
14 (daytime catch in water tunne1)
27th
373
b'9[
(Fazenda Velha) 127
62
m
Grend total 1166 bata of .53 spp.
.aa.
B1ood for sero1ogy and brains, viscera or sa1ivary glands for virus, were taken
from !:lany of the coJlDnoner species. I11oculated mise were kept for 21 days to watch
..

clusive. a.
12 -
for signs of rabies, but no virus was isolated from over 250 bats o The sera oí
558 (including 20 ~l~ from Amapá were tested against 8 uni ts af 10 antigens
by HIj only 21 reacted, as seen below. By comparison wi th African experience, i t is
striking that there were only 2 Group B reactorso The 8 Itaporanga positives include
at least 4 species af bato Six af these pasitives were canfirmed by NT, the ather 2
The 1-fucambo HI posi tive was also confirmed by 1:-TT" but the Jurona
positives were note Tes~ for Tacaribe and AN 67949 antibody have yet to be dane.
Artibeus lituratus
jarnaicensis
spp.
Carollia perspicillata
}'{y"otis albescens
Fhyllostomus hastatus
Sturnira lili'Um
tildae
Vampyrops helleriIndet.
V i r u s
qroup ~ ~roup B GroupC Itaporanga~ ~ Tacaiuma.
1
1
'l'otal 2 2
1 1
2
1
1
1
4
1
1
1
1
1
4 5
The 304 ba~.sera from Amapá tested during the year presented a different
picture (Tab1e 11), probab1y due to their different species composition, being
mainly molossids from houses rather than forest-living bats. Six. af the 7 Graup B
positives had their highcst titre against SLE (one has been identified as
Rhinopl!;r~ pumi1io-CH 1212), the seventh was 1:20 for YF, 1:10 for SLE and
Bus suquara , and there were 13 other sera positive at 1:10 for Group B antibodies,
.
including 3/9 bats ~rorn a single house.
Tacaiuma (1:20) were also found.
Sing1e reactors to Group C (1:10)
3
1
and
All were negative for Guaroa, Maguar{, Turlock

cerine. aboration, th),
13
FIELD STUDIES m ANAPA TERRITORY
In view of the isolation of a new vírus (An 70563-Amapar.{), belonginG to thc
Tacaribe group (of which J\D'lin &. ~chupo viroses, responsible for the Haemorrhagic
Fevers of Argentina and Bollvia respectively, forro part, besides Tacaribe virus),
it was decided to continue the project which has been carried out since May 1964.
~Te continue grea t~ indebted to the ICOMI mining company for their hospi tali ty and
During April there were suspected cases of yellow rever in the POrto Platon
region, which led to s tudies there, however the only virus found was Tacaiuma,
from a pool of ~~ spp.
SERRA DO NAVIO
Capture methods re~ined the same for rodents and marsupials (hardwood live
traps) and birds (mist nets). Some animals were shot. Ba~ were also captured in
oreans in
MOs qui toes were caught on human bait, during the daytime,
general~ at ground leveI, and sent to Belem alive. Sentinel mice were exposed for
an average of 48 hours.
Studies were concentrated at 2 forest stations, that of Rio Amapari or Terezinha
(RA) which lies between the Hospital and the river (approx. 1 }ano wide by more in
.
and that oí c5. The 2 stations comprise varzea and terra firm~o Up to
June, 10 traps were set in each areaj after June~ 10 more were set at station RA,
where tlley were put along 2 trails called "Estrada dij: Cinturinha" and "Trail A".
As the animaIs became scarcer J the traps were moved 10 or 20 m. from their original

,
14 from Jun. 1964 Serra
locations.
At station c5 they were distributed in the forest on each side of the
road which runs through the area.
Birds were captured at both stations, mosquitoes only at RA. Ba ta were taken
various places: RA, surroundings and interiors of houses, caves and culvertso
TabIe 14 shows the animaIs captured and specimens collected for virus isolation or
serological tests, as well as the methods used to preserve the specimenso
It is interesting to note that most of the srnall rodents identified as Neaco~
~anae were captured during August, as shown below, together wi th the captures of
~~ ~! (?), these 2 being the onlyan1lnals so far yielding Amaparl virus:
Neacom.Y'8
~z_o- nrr~
Jan. Feb. M;1r. Apr. ~y
o o 2 2 3
10 2 2 1 6
Jul. Aug. S~p. Oct. Nov. Dec. Total
14 18 53 19 T( 1 5 124
7 17 13 14 8 12 4 ~
During the first half of 1965 the ratio of rodents captured to marsupiaIs was
whi1st in the second ha1f i t was approx. 2 :1. Trap occupancy was japprox. 5%.
~~squitoes identified totalled
\C1? lY
10,521, inoculated in 381 poola. The majority
were ~orophor~ (46%), ~~ (25%) and ',~o~ia (21%). Of ectoparisites, there were
2,354 ~~~ (over ha1f being from ~o~s), 436 Ixodidae (Minly from M9.rsupia1s
and a few other types (see Tab1e 16),
Results -
A total of 178 sentinel mouse groups was exposed, all at station RAo
o ,.J'I ~ ~ I U t! ' ~ \1"'\01. dot f'r""'J' ~ ( ~
_-?/Z ~.~ ~V.~ Q
Amapar{ virus was the on1y virus isolated at Serra do Navio during the year.
virus has been identified as a new member of the Tacaribe group by CF, clearly
distinguishable by NT from the other members. Including 4 isola tions from rodents
and their ectoparasi tes captured in
(2 animals were caught on 23 -December 1964
and sacrificed on 18 Janua~ 1965), we now have 15 strains of Amapari virus from
do I~avio: 7 from the viscera or O~omys- ~! (1), 6 from viscera and

suquar~. quitoes. oa
from blood of Neaco~ guianae, and 1 from a pool of ectoparasites (Gamasidae) -
colJ.ected from 8 ~zo~, which included 2 which produced viruso Table 1.5 gives
the details oí these isola tions o No virus was isolated from sentinel mice or from
CF studies of 16 bats, 4 Neacornws, 18 O~zomys, 23 Proechimys, and
~ --.,.
marsupiaIs of Amapa were undertaken with Amapari antigen.
26
.t
The only CF posi tive
sera were 1 Neaco~ and 2 ~omyso aí 24 sera oí rodents from the utinga recapture
area J none were CF pos i tive o
Reutra~~~~ion testing in Mice witJl~e~ Taca~ri_be- group Agent
The LD50 of 7th passage mouse brain of the prototype AN70563 was approximately
2.5 logs and some mice recovered in alI dilutions. CF testing oí brains oí mice at
each dilution was undertaken to determine the infectivi ty end-pointo Tes ting of brains
aí healtl\"f-appearing mice taken an th~ 10th day fol1awing inoculation showed
the infectivity was 6.7 log LD,O- A N test in mice is under study, using CF as an
indicatar ar infectivi~.
and~2o71og 1D50 in two different tests.
Ama.p=Í were negative.
A homo1ogous hyperimmune mouse serum neutra1ized ~ 3.1
Three Oryzo~ and 12 human sera
Serologica1 testing by HI gave the resu1ts shown in Tab1e 17. In addi tion, some
marsupial and rodent sera were tested for antibodies of the Bunyamwera group and
Capim complex, Tacaiuma and Jurona, with negative results. One bird serum reacted
Turlock (AN 32260).
(Mucambo and Pixuna).
~
Group A positives in fFoe_c~~~~ were for the VEE complex
Among the bats there '~ere 6 Group B positives (l~20) for SLE
(AR 23379), of which 2 also reacted at 1:20 for yellow rever, and 5' at 1:10 for
One other serum reacted at 1:20 for ye11ow fever and 1:10 for SLE and
Bussuquara» and one was positive at 1:20 for Tacaiumao
for B1n1Yamwera group
All the bata were negative
and !-nguari Turloclc and Jurona. These serological

16 Theso ar precarious.
oí nnrsupials, 3inoculated
, results; vaccinated However"
positives are interesting in view of the failure of the virus isolation attemptso
There are at lcast 2 possible explanations for this: 1) inacti va tion of vírus in
organs preserved in glycerine, and 2) disappearance or viraemia due to the orten
long
interval between capture and sacrifice or the animaIs sent aIive to BeIem.
stud1es vere made because of the suspicion of a yellow rever aetiology
fatal human cases.
planting and approxo
These localities were being deforested for sugar cane
400 men were working in them. Operating under contract, these
men were scattered in groups af 10 or 20, living in thatched huts without walls in virgin
recently cut íarest; their canditians aí nutritian and hygiene were extremely
It was the rainy season.
The following s tudies were made: 1) attempts to iso1ate vírus from the b1ood
íebrile patients, from mosquitoes, írom blood or viscera of primates, rodents and
other arboviruses,
2) serological tests with human and animal sera against yellow rever and
collection of information on fatal caseso Eleven human s era
into GMK and HEp2 cultures and 3-to 4-day-old mice (ic
these inoculations were made onlya few hours after collection af ~~e
,
spec~nens. The on1y isalate made was one of Tacaiuma virus, from a poal of
~~~ SPp.
with negative
~10 days before the beginning of these investigations the workers had been
against yellow rever, and therefore alI serological evidence of any
recffi1t occurrence oí yellow rever infection was masked. Dr. EdIm.mdo Juarez,
an ICOMI doctor, had already collected blood from 399 of these workers, almost all
at the tL~ aí vaccinatian, and JI
weeks later he made another collection from 21~1 of

these (2:160 219 of ~~: others. 4
people, for serological testing. Eighty paired sera were tested against the
local strain H 111 or yellow rever by NT and RI testo The rates aí canversian were
43% by NT and 45% by HI.
Further HI tests against ather arbaviruses af Graups A & B
indicated the presence of antibodies against Mayaro-Una (21%), MUcambo-Pixuna (17%),
and 6% against alI 4 of these viruses o
In some cases the Mayaro titres were high
or greater), but except for ye11ow fever virus there were no conversions for
the viruaes tested.
Finally, the histaries af the 2 fatal cases, abtained fram relatives and friends,
did not appear compatible with yellow rever.
~ rivers.
KAYAPO-GOROTmE nJDIAN Smm1 SURVEY
The Kayapo-Gorotire live in the region bet.~een the upper XingÚ and Araguaia
At an Indian Protection Service post at Itagorotire we collected blood from
apparently well-fed and healthy Indians ar all ages (Table 18). Only reactions
1:20 or greater with 8 units of antigen are considered positive. There is no
significant difference between the sexes in the antibody rates found.
most sera were broadly reactive, but with Mayaro titres higher,than the
There were 2 adlllts in which the Mucambo titre was highest, and
ane maIe ar 14 wi th a WEE ti tre ar I :20 and Mayara & EEE ti tres ar 1 :10 anIy~
AlI ather EEE and WEE positives were alsa positive in higher titre to (~
I1ayaro, except for 3 males and 2 females with equal titres for more than
one antigen.
~~-1i: the whole population was said to have been vaccinated for yellow fever
years previously, but all except 8 yellow fever positi'ves reacted equal
,
ar higher titre to Ilheus.

-18
QroupC: one ma1e age 40 reacted to titre of 1:20 with Apeu,
Q!~Q~~: 8 males and 1 female positive, all over 25 years old, 6 of the males
.,
and the female had highest t:Ltre for Moju.
-

g r,
TISSUE CULTURE STUDIBS ON .A.RBOVIRUS~
",",'li
S IMULTANEOU S ISOLATIOH IN MICE AND TO
Duriag tbe ~ear, 1064 samples of flo~d material were tes~ed,
20 attempted relso1atloas. !he material coftslsted
mosquito poola, blood or Y1scer& ot rodents, marsupials, bats and
primates, and b1rd p~asmaso The 562 b1rd plasmas were inoculated
1nto both inrant mie. ~d TO at the same d11u;1oa, but the rem~!1.'_ng
samples vere generall1 diluted 2 to S times a.fter mOUBe iIloculatlon,to
prov1de 8uttlo1ent yolume for the !C 1Roc~at1oa. The ro s'1stema
used were a greea ~loDke1 kidae1 contiauous 1188 (GMK) aad HBp 2 iR
tubes, and prlmar7 chlck embrl0 mono1ayera UDder Nob~e agar (some-
times w1th 400pg ot DEAE/lIlo ot med1ura). 'rb.e tube cultures were
observed mieroseop1cal17 (80x m~~.1t1cat1on) eve~ 2-) days ror
2-3 weeks, and passages made When 1nd1cated.
From the samples 20 T1ruses were 1so1ated 1n m1ce. but onl1
6 or these were slmult&fteousl~ lso1ated 1a TO: Bussuquara, Orlboca,
2 atraias or Guama amd 2 ot MUeamoo.
ot the 20 pool!
poslt1ve ln miee, 081'1 14 vere boeulated into a11 3 'rC sJstems.
aad 1n 3 cases ali eultures were destr01ed b7 ooDtaminatloD, 80
that the 1so1at1oa rate works out at 6/11 Talld testa. OD th.
other hand, aRotber atraia o! Dus8uquara (AR 84600) waa 1aolated 1n
TO alone, and a 8 traia ot I taqui (M. 80465) waa ,reisolll ted 1n 'rc,
Whereaa tne re1so1at1on attempt 1n mice ta11ed. 'D'1e TO poa1tlyee

gether.
og1qu.
20 -
vere a11 1n the GMK 878t.. ~7. ex.ept the MUe~bo 18o1ate8. WhlCh
were both poslt1Te ia 8h1.k embr,o. bu-- ~ OD8 of these waa pos1tlT8
1n GMK, aad tbat waa also poslt1T8 1n BEp2.
IHFLUENCE QF' 'rXPE OF AGAR OVEHLAY ON PL&Q.UE FURMATION
~e fo~o~ t1Pes of agar vere testedt Noble (w1th or
w1thout DBAE). D1tao Ba.to. Oxo1d IOftagar, and Ag&rose (Iadustr1e
Fr...a18e). al~ on &h18k eabr70 monolQ7ers.
Tembe (AR SO1l7). Timbo (AR 41187) aad AH 67949 (bat 18o1ate)
d1d not fo~ plaques uader &a7 ot these agara 1a thls a78te~
In Group J.. }.i1cambo (AR 8 &: jJf 10967) aAd PiXUDa: (AR 35645)
anowed ao difference 1ft plaque morpholog7, clari t7 or dlameter under
Noble agar, wlth O. 100 ar 400;pg/ml. DEAE. Oa the other haad Aura
produ.ed larger plaquea under Noble agar (O or 400 pUml DEAE) tb~
uader the other t7Pea ot agar.
In Group B. Baeto agar was best for Bussuquara, Ilheus and
SLE. Bussuquara prototlP8 AR ~6 produeed plaquea ~~ uader
Baoto agar. Wbereaa stra1D AR 66570 (wh1Oh g1Y8S larger aadclearer
p~aque8 than AR ~ló) a180 tormed p1~e8 under Nob1e agar.
Pre11m1n8r7 testa wlth 7ellow reTer (H l~l) indloated that Ag&roae *&8
best for th18 atraia.
Ia the Chaagu1nola group, the p~aque patte~ was s11ght17
better uader Agarose. 8*rata AR 28873 produeed elearer plaques thaa
AN 41067 & AR 35646, WhlCh sometimes ra11ed to produee plaques

lln
The following Y1rusea only produoed plaqu8s incons1stently,
and When they d1d so, they were fa1nt, badly def1ned and amall
(Wlder 1 mm. dlam.)1 Melao (AR 8033), Chaco (AN 40290). Pacu1
(AN 27326), Acara (AR 27639), Cotla (AS 58058), AN 84785 (Slmbu
and AN 84381 (un1dent1t1ed Ye110w rever Shou1d a1so
perhaps be included ln thls group.
COIofPARISON OF CHICK Alro TURKEY EMBRYO CULTURES
Twenty six arboviruses were tested 1n these systems. The
turkey embryos were tryps1nlsed after 13-16 days incubatlon, the
chick embryos after 10-11 days, otherwise the preparation was
bota cases. 'rhe resulta were as follows:
l-fucambo (AN 8)
Aura (AR 10315)
Ye11ow rever (H 111,AN 78462)
SLB (AN 69768)
llheus (H 7445)
bussuquara (ÁH 66570)
Guaroa (H 12208)
Maguar1 (AR 7272)
Kalr1 (AR 8226)
Tuc Wldub a (AR 278)
lwfe1ao (AR 8033)
Lukun1 (AR 35112)
Oropouche (AN 19991)
-
CE TE CE
+
+ o
+
+
+
+ +o
o
o
o
+
+
+
o
+ +
+ +
+
o
o
o o+
Bujar\1 (AlI 47693) +
Anhanga (Ali 46852) o
Itapor~a(prototype) +
Timbo (Ali 41787) O
Chaco (AN 42217) O
Tacalwna (AN 73) O
Plry (AN 24232)' +
Acara (AR 27639) O
Pacui (AR 27326) O
Marco (AN 40290) O
Jurona (AR 40578) +
Turlook (AN 32260) O
Ir1tula (AR 28813) +
The results in CE were better than in TE, since with CE the
were larger and clearer, the tltres hlgher, and the cells
survived longer. The oe11 yie1d from turke~ embryos was not
s1gnif1csn tly greater than that from chick embryos.
were kindly supplied by Sr. 3el11gmann.
The turkey
TE
+ ooooo+
o
o
+ oo

.
1.
-
22
.AiRBOVIRUSES D GE AND BBp2 CBI.T.R.
Ir1tu1a (.- 28873). Bujara (AB 67744) aad AR 8438t(uaident1f1ed)
were .egat1Yo ia HBpz. the Bujaru a~rata W&a a180 aegat1Y8 but
AI 84381 produced .lear aPE 1a the comtiauous green maDke~ k1dae7
ee~ lia- (GMK). ~arl atraias AX 70$63 (pro\ot}p_). AI 86901
aad AR 81087 vere aegat1 Te or doub't~ ror OPE aad plaque~ ia bo\h
Gl-I{. aad BEp2, 1& the Que ot 'the last .'raia even 1a the preaenee
or 0.1. 1 aad lO~ 00I't1801/81. med1wa.
OOMPUM&:NT FIUTION A.ND '1'ISSUE CULTURE
U~tec.t1oB of A)aaparí (AN 70563) aat~ge!.
Th1s ageat produoes ao CPE 1a HEp2 eells aad aDl7 doubtrul
efreet ia GMK eella, mor does 1t produce pla,uee 1ft ch1ek embryo,
GMK or HBp2 o\Ü tures:. ~erefore Dr. Shope'a te~que for
demonstrat1ag the preseaGe or Amaparí antlgen ia tareGted mouse
braia was adapted to t18sue culture. Ia GMIr. 8ulturee, e1ther tlu,1d
8'10n8 or the Whole inteeted oultur8 atter r~ez~g and tbawlng 2 or
3 times w&e ueed ae ant1gen iR a CP test, w1tn tne following resulte:
Da'18 artariaocula.t10.
Flu1d
T1tre
glJ11d+eella
CPE
6
9121.5
0.1.0
1.0
2.0
3.S
7.5
5.5
.e8....g.aeg.
t
2.$
5.0
I' W&8 eas7 to demoftstrate tne growüb ot the Y1rua 18 culture
bl OP and 1t appears that the bet'er ant1ge. wa. ~at der1yed trom

c lruses peclt'lc.
ver, ver. gcs
both fluld and cells.
- 23 -
Neutra~ization tests using the above teChnique
to determine surviv1ng v1rU8 showed that homologous antlsorum
neutrallzed 210gB or greater of AN 70563. T1trat1on af stra1n
AN 86901 or Amapar{ vlrus ln GI-It oel18 by the same method gave
similar results
Use or combined antl[~en poo12.o
Carey (Nature.G.QQ.J 1024-5. 1963) reported the successful
use of comblned antlgen pools to detect infections by certain
Group D arboviruses in Indla. We therefore tested 48 local
both separatel.,. and combined ln pools, ar;alnst lmrrn,me
fluida, with the resulta shown in Table 19. With rare
exceptions the antiserum titres were ldentical, as a180 wcre the
antlgen tltres.
the antlgen tltres or Group C virusos
were 1n beneral h1gher in combination, whinh 1s possibly dU8 to
close antlgenlc relatlonahips.
3. Teste with material from Labrea. -
No cellular alterations wore seen 1n cul~ures lnoculated wlth
material for attempted v1rus isolat1on from people from Labrea.
on the chance that vlral growth mi[pt have occurred 1n the
absence ar CPE, a11 tlle cul tures trOI!1 orlgirlal inoculatlons or
wore tested by CF, after frcozlng and thawil13 twlce.
a!;ainst the following sera or immune ascitic fluida: (a) Group A
pool (b Group B pool (c the convalescent serum of one patient,
and (d) "normal" human serum. The resulta were all negatlve ar

2u
xmTRALT~--TION TBSTS DI TISSUE CULTURE
5ia.e .er'ata 8aatrad1otor7 resulta vere obserTed ia some
H teste. seleeted 87stems were tested for po881ble causes ot error.
The quaft'itatiye aaal~aea were made b~ the plaque redu.tio. method
ia &h18k embrJo 8ul\urea growa 1D 3 oz. bottles (approx.
aneet area 28 c~). V1rua-serum mixtures were 1acubated at 3700
~or 1 hour and inocula ted 1ft tha Tolume or 0.2 ml. par bottle o
01"OPOU~~.
1. Reprodue1b111'7 ot \1
tlwrat1oms or the a&me T1rus stO8k vere made OB tne
same da7 b~ the same person. fhe d11ut1aaa vere 1acubated at 37OC
for 1 hour to a1mulate HT 8aadltlona aad 1noeulated (0.2 ml.) lAto
3 bot'les per d11ut1oa. Plaque 80uating gaTe the ~o~low1ag
res~t81
Tar1ed trom ~ to 85. 1.8. t1trea ot 4.6 to 4.9 log.
(b) ~ the 10-4 d11ut1oa the aumber ia )0 bot.les ranged froa 1
to 12. 1.e. 4.0 to 4.9 logo It 1a eT1deat that plaque caunta ar
lesa \haa 10 glye larger errors.
2. R.produ.1b111~ or Deutra11zat1oa testa.
Eight mixtures ar the same Tlrua ataok witb tne aame
ant1-Oropouahe 1~~e aseltl. fluid (AP) were prepared, beiag 4
mixtures wl~ 10-1 (25.°00 p.f.Qo) and 4 w1~ 10-2 (2,500 p.f.u.)
or T1ruae Ea&h m1xture wae iAoculated iAto S bot.les. The resul'ing
plaque 8ouata and aeutra11za~1an ~d18e8 (XI) vere &8 tollow81

\dse,
25
dilution gave 19-59 plaques (20 bottles) wi th NI varying thus from
2.6 to 3.11og and (b) the 10-2 di1ution gave 0-9 p1aques (19 bott1es
Titration in the presence oí "normal" animal serum. or ascitic fluid (Ar').
Serum or AF
.
Normal AF
Howler monkey
Human
Guinea pig
Oryzo~
~i'à:eIphis
Virus titre in serum or AF
Inactivated Not inactivated
_._~--- --
5.2
3.0
5.3
5.2
5.3
503
~oval bÜiiifn 4.9
5c
3c
5c.5
4c.7
5. .3,9
4c
'fl1e "normal" howler monkey s ee~ to have been Oropouche iImm.me.
there was no significant difference between the diluents or between fresh or heat
inactivated diluent. In arder to test the effect af the additian af fresh serum
on the neutraIization index, an Oropouche convalescent human serum (Bel 989) which had
been stored at 4ac for more than 4 years was selectedo Fresh hunan serum was collected
a few hours before the tests, which were run in parallel. One tested the irnrmme serum
mixed "Jith an equal volume of the fresh, the other with an equal volume of bovalbwnino
No significant effect on the NI was seen.
~~~~
7he reproducibility of the NT was investigated with 2 viruses of this group.
}ffiE (}k}tlllan strain): 5 mixtures af the same virus stack with serum fram WEE
~~e rabbit 4651 were each inoculated into 5 bottles (total 25 bottles The nwnber
or plaques produced varied froro 9-28 and the max. and mino NI were 2.6 and 201 log.
rfucambo (AN 10967): NI with an immune horse serum varied from 0.4 -1.8 log and
with a hyperimmune AF from 0.8 -1.9 log.
Group ~
Bacto agar was substi tuted for Noble agar in the overlay for Bussuquara virus,
and permi t tJed the production of plaques by this virus in chick embryo cul ture and

1.8 1.4
26 AN NI
a comparison by the plaque reduction technique between the prototype AN~4116 and
strain AN 66570 (which gives larger, clearer plaques), with the following results:
~e~~group
66570-6 inj o
§ mean af 5' tes~
§§ mean af 4 tes~
1:4
1:16
AN 4116
~--
2.6
?
o.s
Vírus
AN 66570
1.9 §
1.3 §§
0.6
strain AN 67744, isolated in 1964 in chick embryo culture from the blood of
a Proechimys, was neutralized (2.7 log) by an AF oí Bujaru (AN 47693 wi th a homologous
aí 2.8 lag.
~_agroup
Both the proto~ SPAN 232 from são Paulo and strain AN 58058 from Belem
produce marked CPE in GI1K cells.
PIAQUE mHIBITIOlJ TESTS
AF Final diln.
AN 58058-5 injo
SPAN 232-5 inj.
1:41:4
1:8
Virus
AN 5805~ ~..?~
2.0
2.0
-
105
-
1.5
In the original Porterfield technique for this test, the fish-spjJ}e beads are
filled by capillari ty on contact ,.dth the surface of tl1e ilmmme serum, a process
vJhich takes up to 1 minute per beado This tas been modified in this labora tory by
using a CF dropping pipette deli vering a drop oí 0.025 rnl. to íill the beads more
rapidlyand accurately. The results have shown ve~ littlc variation bet1~een tests.
In Table 20 are ShO1ffi the results oí tests with viroses oí Group A íound in the
Belem areao It can be seen that WEE and Aura cross-react in one directiono

ined: al
27
ENTOM:>lOGY
This year the entomological research continued along 4 main lines: (a) a
study af the bialagy af ~~,
(b) attempts to obtain oviposition in the laboratory
by species known to be important as virus vcctors, (c a study of the transmission
of viruses to mice by naturally infected mosquitoes, and (d the collection of
irnrnature stages from natural and artificial brceding places in the fores ta 01 IPEAN,
utinga and Serra do Navio J Amapá Terri to~.
In the 3 months February through April, the following breeding places were
Natural
IPEAN 37 4
utinga 61 17
During a S-day visit to Amapá in March, 32 natural breeding places were
examined around Serra do Navio.
In the thatched hut in the lPEAN forest, over 800
mosquitoes of both sexos were reared to adulthood and prescrved with their exuviae
for taxonomic study. In the laboratory, 4 rearing cages were set up as used at the
B 9, B 17 and ~.) taeniopus) released into these from the blower trap catches.
From 5623 female ~ B 9, 3 oviposi tions were achieved. With a rnodification of
the cage, involving placing a gourd under the porous pot, containing organic matter
from the forcst floor, 5 ovipositions were achieved from C.(M.) taeniopus, 4 from
Culex spp. , and 2 !ram Culex B 17, this last in anly 6 days af experimente
-- ~y oí
these eggs were reared through to adulthood, but their maturation was extraordinarily
prolonged in the laborato~ as compared with the thatched hut in the forest.
A family of sentinel mice was placed in each cage daily as a source of blood, and

-28 -
these mice were watched for signs of v1rus infection.
4,367 ~~ spp. , 4 virus isolations were made,
~oJ}1. )taeniopus and 888 ~~ B 17, as fol1ows:
Da te
Virus Strain
From such mice exposed to
plus one each from 3,658
~squito
26 Nepuyo AN 83010
Culex sppo
331 Mirim
AN 83378
Culex (M.) taeniopus
Bush Bush AN 84269
Culex spp.
4
21
27
Capjj!l
Capjj!l
Guamá
AN 84461
AN 84948
AN 85058
Cu1ex spp.
Culex spp.
Cu1ex B 17
This is direct proof that these species are involved in the cycles of these
viruses in natureo
Tables 21-23 give the results aí these aver the year.
There were 13 species
taken by day (0830-1030 hrs.) at Utinga which were not taken by night (1800-1900 hrs.
a t IPEAN on human bait, not surprisingly since these are mainly day-biting species.
From the foot of Table 23 i t can be seen that from Februa~ through April, and again
in .June, October and November, the utinga day catches were the more productive, with
the yearts rate nearly 20% more than the IPEAN night catches.
(swamp forest
The IPEAN V9-rzea
was slight~ more productive than tte capoeirão (secondary forest)
during the second half of the year.
The on1y possib1y significant qualitative
differences between the faunas of the varzea and capoeirão appear to be that the
varzea produced only half as many .9.!-~.spissipes and A?.(S.)nimbu~ as the capoeirão.
These 2 catching areas were about 500 mo aparto

to
29 -
Tables 24 and 25 give the results from Amapá. L~tus pseudomethisticus was
captured there but not at Belemo The preference af !:!ae~ga~~ and Sabeth.es
trees and of ~~, ~rophora, Limatus and \'vyeo~a for the ground is clearly shovm
in Table 25. The capture rate was higher on the ground than in the trees, yet lower
for that month of April at Porto Platon than at Serra do Navioo With daylight
catching time comparable to that at Utinga, only 38% or the utinga annual m~uito
total was taken at Serra do NaviOe The hourly capture rate never dropped below 59 at
utinga (November), whereas at Serra do Navio i t relI to 3 in December and was
less July through September. This does not correlate with the rainfalllevels a~
Serra do Navio, which remained high until September (Table 9).
~~o- JJ~plower Trap
A blo~ler trap of the ~ described in last year's Annual Report was modified
to separate the catch into 4 collecting cage", at 6-hourly intervals. The cages are
mounted in a row above the hood with bait and blower, which travels along rails,
winched into position by an elcctric motor operated at appropriete intervals by a
seOO!1.d clooko Table 28 shows the resul ts by period or capture and species. 'I'he trap
was stopped between 0130 and 0830 to allow for removal of the catch and resettingo
Capture rate shows that the period 0830-1330 is the least productive, 1930-0130 the
The timing should probably be al tered so as not to spli t dusk between 2 periods.
The only species to show up as diurnal by this method was ~tus paraensiso
Comparison of bai~
At 6 mo above ground level in Utinga forest, 6 different baits were exposed on
different days during Janua~ in a blower trap (Table 26). Apart from the mice, the
bai ts used the same cage, so there may have been some carry-over of odour from one
bait to tJ1e next. Human bai t (included for comparison appears to be by far the
mast attractive, cansidering that each catch-day cansisted ar anly 7 daylight

compared Alternative~,
HUrnan shows However, conclusions.
Haney ,
-30"
wi th 24 hours for the blower trap. This could be related to the greater
surface area af human bait.
the blower.
the catchers were more efficient
bai t a ttracted a spectrum or mosquito species almost entirely
different from the other baitA9,even other primates, but it is possible to explain
the lack of Culex by the fact that the hUtMn catch was in the day"time onlyo It"'looks
at first sight as though
that Sabethini can be taken in the blower trap. There ia little to chooae
between the other bai ts as far as mosquito spectrum goes, alI being heavily weighted
the neinly ground-living nersupial Didelphis. It is interesting that the arboreal
marsupial ~~ and the monkeys are apparently less attractive to mosquitoes than
the terrestr1al ch1cken and mouse, ind1cating (a) that an attack rate calculated from
mouse or chicken baited catches cannot be extrapolated to these monkey species, and
(b) that the mouse and chicken are the better trap baits for monitoring arbovirus
activity in mosquitoes above ground level.
attractiveness need to be confirmed by more extended observations.
figures are heavily biassed by .Culex (J.wlanocanion) spp.
these conclusions on relative
The chicken
The rates in Table 27 are more difficul t to compare, as the bai ts were not alI
exposed during the same month, and toa few mosquitoes were caught to permit firm
Only the f!co=e~hi~, mouse and honey figures are direct~ comparable,
and show that there is little difference between the 2 rodents, quantitatively ar
qualitatively, except that ~.?1h~ appears to have selected Limatus paraensis
was the least attractive bait af alI, except the guinea-pig, but the latter
~1as tested in September when in any case the mosquito population reached itiannual low.
The chicl{en rate for October (Table 28) 1s somewhat below the mouse rate for
saroe month (Tab1e 31
whereas the reverse 1s the case at 6 m. above ground

-31
(Table 26), so it seems that a chicken on the ground is relatively less attractive
than a chicken 6 mo highero (In October the chicken catch was not biassed by
r'1elanoconion) . The birds tested did not approach the chicken in attractiveness,
--~
but they died during exposureo The cebus appears to be more attractive than the
other monkeys, and its rate of capture was comparable to that for mouse bait in
the saroe month (Tab1e 31 The lizard (~e- ~~ was onlyone tenth as attractive
in Trinidad (TRVL Annua1 Repor~ 1961 & 1962)0
The preíerence oí ~~~~~ and h(:M:.)ta.~~~~
and B 19 for ~~~. and mouse is clear from Table 27.
for chicken and of Culex
This is not due to the
month of exposure, since E~ B 9 & B 19 were actually more abundant in January
(lihen the chicken bai t wa s exposed) than L~ April (rodent bait -see Tables 29 &
Human and chicken baits attracted the braadest spectrum ar species, but these baits
were exposed for more days than the otherso Over the entire year, mouse bait
attracted 42 species (Tab1e 31)0 As an index of efficiency, in February a chicken-
bai ted Blower trap achieved 6 tj.mes the rate per day af masquitaes taken by a chicken-
baited Magoon trap in the same montho
Feeding percentages froro Table 28 are as follo'~s:
Ae.scapularis
Culex spp.
-'B 7
B 9
B 19
(C.) declarator
(f.f.) spissipes
(H~I~~~
6%
4%
7%
8%
8%
16%
10%
2%
These rates are discussed under the section on mouse baited bloi-Jcr trap catchese

Ae.
catchos,
Mouse bait -hood catches
32 -
Tables 29 and 30 show these captures from utinga and lPEAN. The s~ries
on Table 30 indicate that in both forests catching fell off sharply in the second
half af the year, and this was by a greater factar than in 1964, in which year the
monthly totals were consistently highero Those species that were marked~ more
abundan t iJ1 one year than the other, in spi te of comparable ca tch time, were:
serratus
~Ü:lexspp. ~-
cülex B 7
~rMelanoconion) caudelli ?
~~;}spJ.ssJ.~_s
r;~- 0-1.~Jtaeriiofjus
~Bnson~a venezue~ensis
~ ~ --~~-~-~~ ~~-
Ae. scapularis
C-ulex ~I
cu!éx B 8
"CüIex B 17
Virtua.lq a11 8pccies were more abW1dant in IP;~'Jl catchcs than in Utinr;a
and 10 species were recorded in hood catches from IPEAN which were not found
in utinga hoods this year; on the other hand~ a single C. (Microculex) stonei \-Ias
taken in a utinga hood whereas none were taken in IPEAI~ hoods o
Over a quarter or
alI unidentifiable ~~ spp. caught at IPEAN were taken in January alonej Utinga
did not have this marked peako
The month1y totals for fed and unfed mos qui toes in Tables 29 & 30 are misleading,
as they suggest that about 60% or the mosquitoes taken are unfed.
Examination oi the
species totaIs, however, shows that this is due to Iarge numbers of unfed ~~ spp.
and ~~ B 1, as in the majori ty of the other species wi th combined totals or more
than 20, the number of fed mos qui toes exceeds the number of unfede The ~~
I
mentioned presumably have some other intercst in the hood besides the bai t, perhaps
as a resting shelter.

it oes
-blower trap catc?es,
-33
It has already been shown that mouse bait is one of the most attractive for
(Tables 26, 27). Tab1e 31 shows that the peak biting popu1ation was in
April, wi th a marked decline in the last 5 manths af the year matching the decline
in mouse hood captures 30). TotaIs caught are of course much higher \vith
the blower trap which collects for 24 hours per day, whereas the hood only holds
those mosqui toes which are still resting inside 1fhen i t is closed in the mornlng.
From February through July such large numbers af mosquitoes were taken (in April,
over 18,000 in 27 trap.days t only a proportion of them could be identified in
the time available o needs to be seriously considered whether the removal oí such
largo numbers of mosquí toes has an effect on local ~ numbers and vírus cycles.
The rates for April show that there was not much difference in productivity
between the 2 sites used for the trap.
The spectrum of mosquito species captured
\~as similar wi th both hood and blower, al though ~ B 1 which was taken in fair
numbers with the hood, did not appear in the blower trap catches, whilst the reverse
was the
mouse-baited blower trap during the year, compared with 5 in the hoods.
In spite oí the very much larger numbers oí mosquitoes taken by the blower,
vírus isolation rate per mosquito was less than halí that oí the hood catches
a t IPEAN, al though the ra te per
Virus
EEE
Mucambo
Bussuquara
Apeu
Itaqui
Nepuyo
was almost identical as shown below:
iso1ated from mosquitoes from mouse-baited
hood and b1ower traps, IPFAN forest, 1965 §
Hood
1
1
1
0
0
0
Blower
1
0
1
1
1
1

Actually, Nullipars,
Oriboca
GUaIra
Catu
Capim
Guajara
Bush Bush
Mirim
-34-
Total
Total mosquitoes
Rate per mosquito
Total pools
Rate per pool
o
o o
3
O
O
O
6
2,289
1:382
242
1:40
Blo'ver--
19
15,634
1:823
809
1:43
§ except January & r-ny, when blower trap did not opera te.
the difference in rate per mosquito is barely significant at the
10~g leveI (x. 1.683), but it is conceivable that the time or closing the hoods
(around 0800 hours) selects L~fected mosquitoes, since these could have been taking
their blood meal around dawn after having spent the night ovipositing.
uninfected and interested only in feeding, will have fed at all hours of the night,
beine retained by the blower trap, but most~ leaving the hood before it is closed.
Table 32 sho,vs the effect an the catch af changes in the fan cycle of the
blo1'1er trap. Since the cycle was only changed after 3 or 4 days at one settingg
short-term fluctuations in mosquito abundance severely biassed the 15-minute figures
for Auguste Using only September through December totaIs, the rate per day \~orks out
at 48 for the 15-minute and 39 for the 3D-minute cycle, a difference of nearly 25%
in favour of the shorter cycle. -- Gulex B 7 and B 9 emerge as the active feeders,
achieving 20% ar more within 30 rninutes, and around 1Q% in 15 minutes, whereas
Culex sPp. and the subgenus J.Ielanoconion are the slow feeders on mice.
-L- ---
Ae. -~-
scapularis,
1
4
2
4
1
1
1

er:
on chicken bait, and ~~ B 9 and C. (M.) taeniopus show higher rates on cebus
(Table 28).
35
Thus a 15-minute fan cycle serves the dual purpose of collecting more mosqui toes
than the 3G-minute cycle, and reducing the chance of infection of the bait and thereby
the risk af feedback infectian af the masquitaes caught. r1osquitoes which might
become infected from the bai t are in any case prevented from returning to the
environmento Incidentally, aI though the same virus types were isola ted during the
year from both mos qui toes and sentinel mice, in only 3 cases was the same virus
isolated from both the mouse bait and the mosquitoes caught on it, i.e. January:
Guajara (AR 81105 and CM 7475), September: Oriboca (AR 94240 and CM 8535), and
Catu (AR 95078 and m18629).
There are some discrepancies between the hood and blower trap figures for the
relative abundance of mosquitoes on a month1y basiso Ignoring January and I'BY,
during which the~ower trap was not used at IPEAN, the order of months of the year
in which mosquitoes were most abundant in the 2 traps was: -
IPMN hood IPEAN blower
r-arch
February
JuneApril
July
August
OctoberDecember
SeptemberNovember
-
- --
April
February
r-Brch
June
July
August
October
November
December
September
The trend, hawever, is the same: abundance during the first 6 manths af the
year, a falling cff in Ju1y and August reaching a trcugh in September, a slight rise
in October, then decline in l~ovember and Decembero The hood resul ts show tha t

~ And below, species.
January was actually the peak manth, due ta large catches af ~~ spp.
36
B 7 and B 9, and that the May total was higher than those of April or June,
due to a boost by ~~ spp. again. Cu1ex B 17 and C o (M.) taeniopus
~
and
figured much
more prominently in the blower trap catches, and ~~is probably accounts for the
difference in month1y abundance ratings from the hood resul ts.
The relative
abundance over the year (excluding January and May, when the blower trap was not
functioning at I~~N) or the commonest species, taking the ~~ B 7 catch as unity
in each case, was:
13.0
2.9
2.5
B 7 1.0
IPEAN Blower
Culex B 9
C.(M.)taeniopus
Culex spp.
B17
B 7
Ratio
These figures reinforce the conclusion drawn earlier from the proportion of
fed I~osquitoes found in the hoods, that ~~ sppo
3.4
3.2
2.5
2.4
1.0
and B 1 have less interest in
the bai t than in the shel ter afforded by the trap, and that therefore the hood
would take more, since in the 1[o1ier trap any mosquitoes merely resting beneath the
eaves woulc; not be blown into the collectinB ca~e.
Perhaps ~~ B 1
is not taken
by the blo"!-ler trap bec:?use its approach to the ~rl~.t js from the side rather than
and it never enters the danger area.
C.(M.) taeniopus
perhap5 the rea50n 50 few
are taken in the hood is tha t they prefcr to leave the bai t
immediately after feeding rather than rcst close by under the hood like the other
These are obvious fields for experimente
~~use ba_i~ ::!';:inidad Na 10 trap
During January~ 6 of these traps were run for 20 days with screened bait,
to prevent reinfcction of trapped mos qui toes in case the bai t mice became viraemic

ork. ar,
-37
from an early biteo The screening diminished the attractiveness of the bait so much
that on:Ly 14 mosquitoes were caught. During February the screening was removed, but
in 54 trap days on1y 350 mos qui toes were caught (6 per trap day), so the use of these
traps was discontinuedo The traps had been modified from the 1961 TRVL design by
the addition of a canvas top and equatorial band to preserve the nylon netting from
wear a.nd tear.
~E~- bai t " M3.goon trap
A modified 11agoon trap was run wi th chicken bai t for the first 2 months of the
with the results shown in Table 39. In the saroe 2 man ths af 1964, wi th twice
the rainfall, the l~te of catching was 8 times greater, yet 5 fewer species, and
In February
1965 a chicken-bai ted b1ower trap took ~S times the n\unber of. mos qui toes daily taken
by the l!ágpon trapo
~~.t.r.a.p. .c~~~
These were run from 1800 -2400 hrso, roughly once a week, in 2 different
localities in the forest, alternated between catches. Table 34 shows the results.
There was the same September low as in the mouse-baited catches.
The follovdr.B
mosquitoes 1íere not captured by any other method: Psorophora cingulata (only 1 caught),
~edornwia squamipennis (a cano~-haunting bird-feeder), Uranotaenia lovdi and
Uogeometric~ (amphibian feeders)o . AlI males caught were separated for future
i
taxonoroic
The number oí hours catching per month with the light trap is not adequate to
permit quantitative comparison ~]ith other methodso
. ..

,c. Q.~~~c~~e~ for foliage
~ontrap catches
-38 -
Tables 35 and 36 show the resulta oí theseo
and U.leucopterawere not caught by any other methodo
~ficroculc~) B 1 and B 2,
tree-hole breeder), ~ranotaenia hvster~,
l~s ~ t-

h. ,
.39
suction trap took more males than fema.les, whereas the light trap did
the reverse, at a much lower rateo Peak light trap catching rate was in December,
wi th a smaller peak in February; the suction trap peaks were October, followed by
However, since the 2 types of catch were made on different days, in different
forestG, they are not directly comparable.
-Precipitin
tests on blood meals
Precipitin test resulta on blood meals of mosquitoes and Phlebotomus captured
near Bel.em have been recei ved from Dr. Tempelis of Berkeley:
Negative
!.rs.rsu pial
Bird
Human
Rodent
Bovine
44
20 (a11 mosquitoesj 8 were Cu1ex B 9) .
18 (a11 ~ except for 3 Aedes serratus) .
5 (3 from r~sonia venezuelensis, 2 from Aedes
3 (ali Cu1exy--
1 (Cule~)
rõ6'
ar the 19 H11ebotomus meals tested, 7 were positive for unidentified maJ!1In8.1,
and the rest were negative to everything, probably due to unsufficient bloodG
Ticks
f~n attempt was made to rcar out engorged sub-adult ticks found on birds, by
Trapido's technique. Out af 35 callected, and kept individually in 3 x 1 inch
tubes over moist sand, 8 died after 9 da~ and alI but 4 were dead by the 14th
the re!rJa.inder dying soon after without moulting. Apparently the trauma af farcible
rcmov41 from the hosts (which were bled and then released), caused this mortalityo

, poo1
40
PARASITOLOGY
Incidental to the rodent and bird recapture programmes, blood smears were made
at the time of bleeding for virus and serology, and a number of paras i tes ~Jere found
in these (Table 39)0
!~o~p~~
Vírus stocks for yellow fever (H 111
MOUSENEUTRA LIZAT ION TESTING
Ilheus (H 7445), SLE (AR 23379) and
Bussuquara (AN 4116) were obtained by the ic inocu1ation of 3-day-o1d mice with 10%
suspensions of mouse brain in bovalbumin, and the collection of the serum or brain
at 96 hours after infection for H 111, or 72 hours for the other viroses.
In an effort to interpret the specificity and sensitivity or ti".e N test using
serum vírus íc, and brain vírus íp, wíth the 4 group B agents known in Erazil, the
follO\fing 13 seTa were selected for testing in tl1e 1:8 dilution: seTa 1 tm-ough 4
are hyperimmune mouse ascitic fluids for yellow fever, Ilheus, SLE, and Bussuquaraj
sera 5 through 1 are believed to be yellow rever immune either by history or HI
pattern, including a human with a reliablc history af previaus 17-D vaccinatian, a
oi sera of monkeys PR 319 and PR 322 shot during Janua~ in Utinga forest, and
monkey PR 316 from the same forest; sera 8 and 9 fram humans of Gameta believed to be
Ilheus imzmme by the HI pattem; plasma and senlJll 10 and 11 believed to be SLE ilnmune
repres en ting pooled Q~ragypa (black vul ture ) plasmas AV 6306 and AV 6310, and a serum
fram a human resident af Cameta with SLE RI patternj and sera 12 and 13 fram fFae~himya

0
ination,
RO 5209 (several bleedings pooled
41
and RO 5157, be1ieved to be Bussuquara immune by
considerabJ.e experience indicating that Bussuquara commonly infects these rodents.
Table 40 lists the results af yellaw rever, Ilheus, Bussuquara, and SLE N tests
(Strains used given on Table)o Serum 7 , a monkey from Utinga, and s erum 8, a human
,
from Cameta, would appear to have had previous experience with both yellow rever and
Ilheus viroses o The yellow rever brain ip test lacks specifici ty in group B unless
~ ~~I of ~ 2.5 is considered as the criterion for positivity instead af tr_e classic
figure af ~ 1.7. Fbur non-group B mouse hyperimmune sera were included with LNIls .of
0.2, 0.4, and 1.3 indicating that the ye11ow rever ip test group B resu1ts are
group specific, and that reactions represent heterologous antibody. This test should
be excellent for measuring group B antibody.
The brain ip tests with Ilheus, SLE, and Bussuquara viruses retain their
specificity to a remarkable degree and are as sensitive or more sensitive than the
serum ic teste It would seem advisable to adopt the ip test routinely when dealing
with SLE, Ilheus, and Bussuquara,
yellow fever
Yellow rever strain H 20228 from a febrilc human from Vigia, taken 2 days after
was used to test for post-vaccination antibody. This strain;
after the 5th passage, produced a viraemia in mice of 305, but when used as a
haemagg1utinin showed no advantage over strain H 111 in a comparative test on human
sera from Amapa.:
1:20
HI titre
1 :40 1 :80
H 111 60 § 152 45
H 20228 61 155 49
4
§ NQ af sera pasitive at this titre.

-42-
The sera of 495 people resident in Amapa were tested by NT using serum vírus
ic in 4-day-old mice; there were 398 first bleedings and 97 second bleed1r'G~:
Bleeding
1st
2nd
Positives
83
.57
Conversions
-
39
From these resulta it appears that many oí the HI positives among these gera
due to heterologous Group B antibody, and inspection of the detailcd resul ts
supports this view -~ ar the positives had a higher titre with Ilheus antigeno
Changu.irJ.o1a group
In 1964 we were unsuccessful in producing a serum virus preparation for NT with
(AN 28873). In 1965 we tested brain material of various passages of this strain
stored at-6ooc in different preparations, with the following results:
?!:aterial Date prepared
vJhole mouse
r.yophilized brain
Erain 1:5 bovalbumin
" " "
" " "
l~ole mouse
" "
" "
1205.64
12.5.64
12.5.64
6.3.64
1e.3.65
21.5.65
28.5.65
102.65
Date titrated
--o
same20.5. day,64,64
20.5.
1.2.
30.3.
same
same
same
10, L.6
.5
3.5
.5.5.6.602
2
3
.5
.5.404
.5
therefore, recently prepared mouse brain, at most 8 days after collection,
the folloi~g resulta were obtained:
Virus
AN 28873
AR 35646
AR 41067
AR 54342
AN 27639§
100 §§
O
003
003
ImImme Asci tic Fluid
AN 28873 ~~6k6 AR U067 AR 54342
2.8 a
'O";B:b
002
0.5
§ Acara control
§§ neutralization index (log LDSO)
(a) mean of 4 testa
(b) mean of 2 testa
(c) mean of 3 testa
.65
,65
day
day
day
100
2.3 a
-o-:(}b
003
0.8
0.7 c
2.1 b
~
0.5 o
2.8

-43
When the test was repeated by plaque reduction in chick embryo TC (with
Agarose plus Colems.n medium), the differences were even clearer (see below)o The
ascitic fluids used were the same as for the mouse test (final diln. 1:16), but they
were heat inactivated at 56QC for 30 mine
VinlB_.- AR 41067-7~.
AN 28873
AR 35646
AR 41067
1.8 b
õ:J;b
0.3 b
(o) as above
002 c
109 c
'O:Ic
o c
003 c
~c
The AN 28873 plaques were clearer than those or the other 2 strainso are
awaiting the arrival oí antisera to other viruses oí the Changuinola group, in arder
t~ carry out further tes ta o
Using AN 28873 in challenges of 20 and 200 1050' 27 ~o~ sera from Jtinga
and 24 from the Belem-Brasilia highway (whence carne the Q~zo~ and ~leboto~~
which yielded these strains) were tested in mice but none were protective.
This strain, isolated from a sentinel mouse, has a close relationship wit}
Acara (AN 27639) in the CF test and with Capim complex antiserum and the Belem
~ wí
strain of 3ush Bush (AN 20076) in the HI testo A serum virus stock was obtained
from infant mice bled 42 hours after inoculation with 10% infective mouse brain,
and compared with a serum stock of AN 27639, with the follo\dng results
(neutralization indices): !nmIune Asci tio Fluid
Vírus--
AN 84381
MJ 27639
409
'a:4"
002
5.5
Capim Capim
Complex ~~
1.9
0.4
089
-
Guajara
AN 106~
0.9
002
Bush Bush?
AN 20076
309
0.3
It is thus clearly separable from Acara by NT, and closest to Belem Bush Bush
or those Capim complex viruses tested.

eek agança ~~ lds, deed re ble 1er
44-
IABORATORY CONTAMINATION CFmCK
With the isolation and passage of so many strains of virus at the same time
in this laboratory, it is to be expected that cross-contamination will occur, and
!rom time to time isolations have been discounted when other evidence tended
to cast cloubt on their validityo But a useful check that cross-contamination is not
comm.:>n
be made by looking at the viruses isolated from sentinel mice in adjacent
boxes (Table 41) and from material inoculated L~ series 42)0 It is clear that
adjacent mouse groups do not yield the same vírus any more often than might be
expected by chance. There does not seem to be any frequent spread or infection
on the shelf or on the inoculation tableo
REroRTED EQUllm .ENCEPHALITIS ll.J BP..AOOJÇA
In 1960, EEE virus was isolated from horses and ~s ~e~or~ch~ mos qui toes
in the Bragança regiono
Intensive studies during 1961, 1962, and again L~ late 1963
the initial heavy rains each year, but no return oí EEJ activity was detected.
This year, after reports af equine encephalitis were received durin!
in I~rch, a field team went to Bragança to investigate.
the third
l~o encephali tio horses
'~ere faund in spi te af a hause to hause canvas by canoe over the recently flooded
however nW:1erous first-hand reports "lere received or equine deaths from
"mal de rodas" during the previous ~'1ree weekso Mosquitoes were said to have been
abunctant but to have disappeared coincident wi th the flooding and wi th the
disappearance af clinical disease in horses two weeks beforeo Almos t no mos qui toes
evident at the time of the investigationo
It was concluded that probably
equine encephali tis acti vi ty had returned after an absence af 5 years in the
region

iolof:;f
45
CATTIE DISEASE
In Ju1y we were asked by the Serviço de Defesa Sanitária Animal to investigate
sickness and death in cattle in the Belem area following routine rabies vaccinationo
Clinical history was of onset 2-) weeks after vaccination, with 1088 Or appetitc,
axcess1ve salivation, ~rsensitivi~ 1n some ca8jsand paraplegia in 5/64 animaIs
examined.
Six animaIs presented with rever, but foot-and-mouth disease, type
Rezende, was identified from tongue scr4pings oí one oí these, by the Centro Pan-
americano de Febre Aftosa in Rio de Janeiro.
five animaIs died o
tis sue from one co,~, brain material from 2 and vis cera from 3, plus the
sera of 95 cows were inocula ted into baby mice and guinea-pigs, also the brain and
vis cera of a sick goat.
BHK 21 ce11s, without effecto
one beíng identífiable as rype O.
The resul ts aí 57 blaod caun ta were:
The goat material was also inoculated into GMK, Hep 2 and
Two isola tions or F & M virus were made rrom blood,
Hypochromic anaemia
Leukocytosis
Leukopaenia
12.3%
33.3%
10.5%
results af testing 140 cattle sera by HI with 9 antigens were 12 positives:
EEE 2, WEE 2, r.fucambo 2, Pixuna 1, Ilhéus 1, Tacaiuma 1, Maguar! 4 (one was positive
against both WEE & Tacaiuma)o Six of the positives were from a single dai~ herde
-~ll ti tres were in the range 1 :10 to 1 :20 on1y.
Pathological findings in the fatal cases were oí encephalitis of the lymphocytic
type (2 brain examinations) and sarcosporidiosis (4 heart examinations).
revealed only non-haemolytic staphylococci and streptococci, and Gram positive
bacilli which turned out not to be B. anthraciso
The suspect rabies vaccines were inocula ted by us into mice, rabbi ts and guinea-
pigs, without producing any effect; they were also checked and cleared by the

just trabeculae.
46
veterinary laborato~ at Recife. Our conclusion is that there was no viralor bacterial
cause for the outbreak, but that in view of the pathological finding of encephalitis
in 2 cases, a delayed allergic reaction to the -~ccine could have occurred.
r-DNKEYS
As the supply aí unused sentinel cebus monkeys ran out in June, a new batch
was purchased from the regular supplier at !vL:iracana., whom we were lucky to contact
before he moved to another district. Out oi a score purchased, 17 survived the
ettling-do~m period and were bled to check on their freedolftfrom antibodies, together
1~th 7 survivars ar previaus batchese Their sera were tested by HI against 4 units
of -each of 25 antigens. All the ne\vcomers were negative throughout, except for one
positive to Maguar! and another to Bujarú, both at 1:40. aí the old-timers, !1! 366
whiéh had contracted EEE in utinga as a sentinel in Februa~ 1963, still had EEE
antibodies at 1 :20 1n .August 1965, but no hetero1ogous Group A antibodies; ffI 391
"lias the LA.N sentine1 which contracted ye11ow rever in December 1964, and had a 1:40
titre to that in August 1965, being negative for Ilheus and Bussuquaraj and }rr 363
and MY 387, \ihich as sentinels had been infected in February and Septernber 1963,
respectively, with Marituba, were negative < 1:20) for }~rituba and 6 other Group
antigens
Cebus monkeys MY 372 and MY 397 J both females which had been kept in the
1aboratory since 1961 and 1963 respectivcly, died during the quarter, and were found
ta have very brittle banes, the marraw cavities af the long banes lacking crass
The condition resembled the cage disease seen in some grivet monkeys
kept at Entebbe, Uganda, in spite of the feeding there of a well-balanced diet

l e,
47
The histopathology in these 2 monkeys was not notableo
Since October we have purchased some 50 ,Saimiri sciijreus, mostly from
Island, and 3 TaIOOrin monkeys.
Seven ar the Saimiri died ri thin 3 days ar
having been damaged before arrival at the laboratory, and over 50% of
the remainder died beíore the end oí the year, possib~ due to lack oí proper
accommodation, but this species appears to be ve~ fragile. have had survival
as live bait in mosquito traps in the" forest.
mUSE COLONY
During August and the beginning of September 1965, it became imperative
to introduce some changes in the management oí the rnouse colony in arder to try
a leveI oí production more adapted to the requirements oí the laboratory.
Two chie! problems were tack1ed initially from an experimental angle in Bioterio 3.
'I'hey concerned: the morta1ity in the breeding sector which showed progressive
signs of build-up, and 2)the declining output in litter production and in litter
Mor~li ty
from autopsies examined at the Deparbnents of Pathology and
Bacteriology or the IEG revealed several possible causes or death, some or them or
an enzootic character. Salmonel~ spp.:
with a wealth aí enteric agents,
isolated. Furthermore, a 40% infestation with a Cestode of the Hymenolepis
genus was faund in dead ar sick mice. The intermediary host appeared to be a
Dermaptera of the Forficularidae family, an abundant commensal in the mouse boxes,
several specimens af which cysticercaid larvae were faund.
Besides these enzootic
situatians, a temporary outburst af pneumonia with massive lobar condensatian

started in mid-December.
48
The most urgent measures were directed against water pollution which was found
to be high. Difficul ties in the procurement of fil ters for each Bioterio delayed
until the end of the year the installation of alI the desired filtrationj trouble
also arose from the bad quali ty and probable rough handling of the candles, wi th the
result of some unexpected seepage through some of them. The highly indicated
sterilization oí water bottles in verdunized baths also sufíered much delay.
Other precautions regarding the actual handling of mice were recommendedo
The use of seven-inch forceps frequently dipped into a sterilizing solution and, in
general, the avoidance of hand-mousQ contact were stimulated constant~o
the sex-sarting aí iníants requires handling; hawever, this aperatian lascs much aí
its danger by the practice of frequent hand rinsing.
Unfortunately,
During September, October and part of November, radical elimina tion of diarrhoeic
mothers and their broods was enforced. This was especial~ hard as very often a
heal~ contact had to be sacrificed. Since mid-l~ovember, though, diarrhoea has
disappeared from the breeding sector. It still ia observed, however, in the roam
where inoculated groups are kept for daily inspection and more especially in mouse
groups kept for long periods. This room has not been provided with a filter, as the
material is expendable. This observation tends to confirm the utility of water
filtration as a preventive measure against enteric agents.
Progress of some sort seems to be on the way. The ~enolepis infes ta tion \-lhich
in September appeared to b~ o! seconda~ importance is the chie! concern now. It
appears to affect mostly lactating femaleso Exterminatian af Dermaptera, although
active, is mainly mechanical; the wooden cages and wooden stands in use, \~th their
innumerable cracks and refuges make of the disinsectization of the colony a difficul t
tasko
In November, a progressive deterioration in the supply or wood shavings utilized

io. d
49
litter (another possible íactor oí mortali ty due possib~ to toxic resins)
their substi tution by rice ~usks.
are provided by a rice mill.
husks are collected in bins before they have a chance to touch the ground, as
they are blown out at high temperature from the exhaust pipeo They appeared at
once to constitute a drier, easier to handle material, not as liable as the shavings
to be contaminated by wild rodents o
Attempts at increasing output
The number aí breeding íemales had to be reduced in September in arder to
diminish the contact factor and the crowding. On October 4th, two boxes were added
to the daily groups of ten breeding boxes, and on October 23rd two more.
raised the daily ins talled breeding íemales to 112 wi th 42 males. Following the
apPearance
observation that the/~R. oí litters has a peak during the six or seven days after
first litter (19th day of mating) and fol1owed ~~~~~ by a sharp decline,
it was thought advantageous to withdraw alI the femaIes from the breedin boxes
days.
This was later reduced to ten days.
the tempo of mating is increased
and a more effective utilization of the space and boxes available is achieved.
the 10ss in production due to the preventive rneasures is partia11y compensated foro
Now that the enteric infections seem to be under control, i t is planned to
ta11 the original number of females in the boxes o Together wi th a smal1.
increase in the number oí boxes eaçh day, compatible with the space available, it may
be possible to reach close to the requirements without the installation of a new
Bioterio 1, after the first observations in Bioterio 3 has been following the
procedures wi th similar resul ts.
1 ShOlffi the month1y percentages of mortality in the breeding sector.
peak that builds up into mid-November reflects the sacrifice of sick or suspicious \

50 -
females and their contacts o The increase at the end of December was apparently due
to the pneumonia episode. Moreover, heavy rains which started at the end of
December seem to have produced a more intense water pollution in the city's network
of distribution. It must be kept in mind that Bioterio NQ 5 which provides breeding
stock to the others suffered a delay in filtration until the end of the year.
2 presents the number of babies per litter, an important consideration in
the output; it depends on several factors which , uníortunate1y, escape our control
in the present conditions. One af them may be heredi tary o Selection is ve~ difficult
to achieve because the use of litters for inoculation entails indiscriminate
Also lack of space and shortage of mouse boxes do not allow for
individual caging af pregnant females, which wauld have the added advantage aí
preventing, to a certain degree, cannibalism. Another difficulty resides in the
deterioration of the food supply by prolonged storage under adverse conditions
temperature and humidityo An attempt to compensate for this deficiency was made by
increasing the ration oí whole rice. Figo 2 sha1vs that, in spi te aí a general
improvement in the conditions or Bioterio 3, no substantial change has been obtained
in the fertilityas represented by litter size
Encouraging resulta appear in the rate of litters per mated females, as seen in
This may be the consequence of a certain degree of selection obtained by
the wi thdrawal of the females from the ma tinE boxes after ten days o There is a
definite trend toward the 60% bracket starting at the beginning of November. Two
devia tions, one a t the end of October and one in mid-December remain unexplained,
total number ar babies is alsa a cause ar aptimism (Fig.4). There was a
rioe, as expected, each time that the number of mating boxes was augmented, on Oct. 4
and 23, but the numbers seem to exceed, slightly, the expectationso Here also,
the trend is toward a stabilization around the 400 value (5 days mean). This result
was obtained in spite of the reduction to 8 of the number of females per cage. It is
hoped that the restoration to 10, now deemed possib1e, may produce a significant
.
improvemcnt. This can be attempted, however, an1y \-Jhen ful1 assurance af contro11ed

Remary.s
..
-
51
supply and other infection sources is obtained.
1. The measures applied in Bioterio 3 were later introduced into Bioterio 1,
wi th comparable resul ts .
The return to the original nurnber of females per ma ting
box will be experimented wi th in Bioterio 3 and, according to the resul ts,
Bioterio 1 will follow, or keep the present status.
2. As this Report is assembled, the trends appearing at the end of tbe
year are being confirmed, and tr.e.graphs of litter rates and number of babies
increased production.
babies per li tter, and a small reduction in mortali ty.
There is even a slight upward trend in the number oí
HISTOPATHOlOGY
The following animals were exaJTlined durir.(1' the year:
Iaboratory
38 adu1 t mice
36 infant mice
17 guineapigs
4 monkeys
~~_tE-C?!ogy of wild animals
Wild Other
67 bata
31 prima tes
19 rodenta
10 turt1es
7 edentatcs
6 marsupia1s
3 others
13 ungu1ates
5 carnivores
2 others
The liver, kidney, heart, lung, and in some cases oth.er viscera, were
exaJnined from 8 rodents i;rom Amapá which had yielded Amaparl virus o Four had
hepatic lesions, but these were of a focal, probably parasitic typeo Three had
focal lesions of the IrtY"ocardium, 1-1ith inflaITU11ation and degeneration.
EIeven rodents trapped in the I~~I~ & Utinga forcsts, alI negative for virus,
examined o
-
One had hepa tic cirrhosis, chronic pneumoni tis and ~ocardi tis.o

52
Thirty animals shot during the utinga yellow rever investigation, alI negative
for virus, were exarnined for li ver lesions; all weDe nega tive o
lIistopa tholo~ of sentinel mice
virus strains:
Brains were taken !rom sentinel mice from which were isolated the folloidng
Group A:
EEE 1
Group c: Caraparú 4
Oriboca 3
l'!epuyo 2
Itaqui 1
Group Guama:
Group Capim:
,
,
Guarna 8
,
Catu 2
,
Moju 2
Capim 1
.I
GuaJara 1Bushbush!l
Encephalitic lesions were found in alI. In addi tion, the EEE-infected mouse
showed perichondrial and periosteal lesions as in mice infected '-1ith }!ayaro virus,
and perivascular ~ocB.rdial and pulmonary degeneration. The slides of EI':E; mouse
mater::i_al from 1964 were therefore reexamined, and 3 were found to have lesions of
the same tY!~. Oriboca and Nepuyo were found to cause hepatic lesions, which were
extremely focal in the case of r~epuyo.
~~rlln~tal ~thology in mice
~10 newborn litters were inoculated ic & ip with AN 20076 (Bushbush?) virus
Only encephalitic lesions were subsequcntly found.
Tnirtj--three sick adult mice from the breeding colony were examined, and hepatic
and pulmonary lesions of a necrosuppurative tJ~e were found, probably of bacterial
origino
Four cattlc brains WC11e examined, and 2 had lymphocJ~ic encephalitis, 1-1i th
peri vascular mononuclear infil tra tion. One aí 5 guineapigs inaculated with material

is: sis:
from these cattle died 1'dth focal degenerative neuronal lesions.
53
The following ~uman material froro sick people froro Labrea was studied:
a) Maria Raimunda: visceroto~o
massive diffuse liver necrosis).
b Maria Rita: liver puncture biopsy.
regenerative phaseo
c) r.~ria de Consolação: necropsyo
acute yellow atrophy (actual~
virus hepa ti tis,
acute yellow atrophyo
Guineapigs inoculated ,.ith tissues, blood and urine froro these cases were
negative on examination for leptospira or 1.esions athributable to viral infection.
A female ~~~ monkey (histol.nQ 658) which died after 3 months as a
household pet was found to have a systemic infection with toxoplasma.

Robert"E.
54
BELEM VIRUS UBORATORY STAFF.. 196)1
Shope, M~.,Rockefeller Foundation Staff Member Jan-Jun.
John P. Woodall,~D.,Rockefeller Foundation S}aff ~ernber Mar~-Deco
Gilberta Bensabath,M.D.,Serviço Especial de Saude Publica All year
--- Virus
Joaquim Medeiros Contente
Lucy da Silva Pereira
tily Toda
~eroloQ':
Amelia Homobono Paes de Andrade
Lindomar de Souza Vasconcelos
Maria atavia Savino Vilhena
.Entomology
Amazônia Toda
Emanuel Nazareno de Frei tas
Maria Auxiliadora Amorim Barra
Clinical Patholo~
Field
Guilherme Br::f.gidO Nunes
Sebastião Fernandes de Oliveira
Raimundo Benedito da Silva
Carlos Marques Cabeça
Glassware and Cleaning
.--
Milton Costa Silva
.,
Màr~a de Nazare Gomes de Oliveira Castro
Jorge Maia do Nascimento
AlI year
AlI year
AlI year
AlI year
AlI yearAlI
year
AlI year ~.
Ali year
Ali year
Ali year
Ali ye ar
Ali year
Ali year
AlI year
AlI year
AlI year

Animal --- attendants and cage washers
Raimundo Cunha Mendonça
RaiImmdo Henrique Cavalcante
Antonio Alves Pereira
Oswaldo Vaz da Silva
Pedra Batista da Silva
Sebastião Gomes de Oliveira
Carlos Gomes
José Batista da Silva
RailnlUldo Pereira da Silva
Afro de Aquino Borges
Ivo Ferreira dos Santos
Crizolindo Andrade Coutinho
Raimundo de Oliveira Ribeiro
Pedro Alves Cavalcante
Oswaldo Trindade de Figueirêdo
Raimundo Rodrigues Pinheiro
Francisco de Souza Pereira
Rield service Oswaldo Cruz Insti tu te
---
§ (
(
(
§§
Arlindo Pinto de Souza
Joel Pinto , de Souza
Heber Jose de Alencar Loba to
(Juramir Barbosa de Oliveira
(Antonio Francisco Pires Martins
(Francisco Ferreira Ramos
(Antonio Santos Lima
(Enéas Francisco da Silva
(?-nnuel Santa Br!gida
Chauffeur and field
55 -
All year
Ali year
Ali year
Ali year
Ali year
Ali year
AlI year
Ali year
Ali year
Ali year
Ali year
Ali year
Ali year
All yea.r
Ali y'ear
Ali year
Ali year
AlI year
AlI year
AlI year
AlI year
AlI year
Ali year
Ali year
AlI year
Ali year
Geraldo Pereira da Silva AlI year
~C!~~
Aracelli l'~ria de Souza Cos ta Ali year
-RF
Bernardino
,
Ferreira da Silva
Lourival Vaz da Silva
Orlando Vaz da Silva
~oel Vaz da Silva
All year
Ali year
Ali year
Ali year
§ paid by roc (Conselho Nacional de Pesquisas) -roc only pays for 1/2 day
§§ FSESP payrol1

Apr.
Name Institution
---
J.Fonseca da Cunha
LóMoITis
W.Dyal
A.AóMedeiros
T8M.~~ck
J.Do1-fi.llar
W óG .Do~!f'.s
RóWéDickenan
C.LoSteph.en
O.Correia
Oólliblbock
A.Young
JeAóBottino
J.S.Neiderhauser
K.Lõ Turk
~B.Schaefer
F .f..Neva.
JesSe D. Parki.nson
H..G.Souza
J.Sequeira
EóAlmeida
R.B.Tesh
D.E.Carey , ,
C.Serie
H.Grillo
PóS .IIumphrey
S.S.Humphrey
D.S.Soleau
TóEolDvejoy III
H.Trapido
C .E1 ton
R..Lainson
R.Hubner
-56 -
VISITORS -1965
Insti tuto Oswaldo Cruz ,Rio de Janeiro
CDC, USPHS,Atlanta,Ga.
CDC,USPHS,Atlanta,Ga.
CDC,Field Stno,Kansas Ci~,KansaB
Smallpox uni t,CDC ,A tlanta ,Ga.
Smallpox Unit,CDC,Atlanta,Ga.
RF,New York
Cornell UoMedical College
US Consulate,Belém
,
FSFSP, Para
Univer8ity oí Amsterdam,Netherlands
New York Times ,Rio de Janeiro Bureau
Faculdade de Medicina Veterinaria,São
RF,Mexico'City
Corn~ll U., Ithaca, NY
Paulo
Inst.of Marine Resources,La Jolla,Calif.
Harvard School of Public Heal th,Boston,Mass.
Dep~Scientific Affairs, Pan American Union
Dep.Scientific Attairs,Pan , , American Union
Universidade do Para,Belem
FSESP,Belém
Peace Corps, Recife
RF,cMC Hospital,Velloee,India
Insti tut Pas teur, Paria
Conselho Nacional de Pesquisas ,Rio de Janeiro
US National MUseum,Washington,DCArl1ngton,Va.
Concord,r-ilss.
Peabody MUseum of Nat.Hist.,New Haven,Conn.
RF,Cal1,aolombia
Bureau of Animal Population,Oxíord University
London School Hyg.&Trop.Med.,London
Berlin
Date
--~
Jan. 2.5
2.5
2.5
2.5
2.5
Feb.11
l-Br. U
23
24
8
23
28
May 16-Dec.
18
18
18
18
28
28
28
Jun. 10
14
18
19/Ju1o14
22 .
24/Sepó
24/Aug.
24/Augo
24/Aug.
23!Jul.l
23/Jul.l
Ju1o 8
12

57
Name Institution
Date
JóC.King
M.V.llilovanovic
Eóde F.Monteiro
H.S1ck
H.A.Sissons
R.SoConard
W.SB.ul
F.\ol.Saul
J.P.Rembrand
A.Rodrigues Filho
K.Someswara Rao
R.lopez Fabrega
A..Villas Boas
AóVó}~tos
PoS.Freire
M.Frione
R.E.Stowell
Inst~Oswaldo Cruz,Rio de Janeiro
Inst.Os\~aldo Cruz,Rio de Janeiro
MUseu Nacional,Rio d e Janeiro
Royal National Orthopaedic Hospi tal, London
FJnporia,Kansas
Emporia., Kansas
Emporia,Kansas Escola de Portugues A e Or1entaçao,Canp~as
.'
Faculdade de Medic~a,Univ.do Para
Oficir.a Sanitária Panarnericana ,RiodeJaneiro
National Bulk Carriers Inco,New York NY
FSESP ,Rio de Janeiro
FSESP ,Rio de Janeiro
FSESP,P.io de Janeiro
Milan, Italy
Armed Forces Inst.Path.,Washington,DC
Aug.17 19
19
23
31
Sept.3
3
3
3
Octo 12
12
20
Novo 16
16
16
23
25

Group
or
complex
-58-
Table 1
Name of virus, Belém prototype, and source of isolations,
1954 – 1965
Belém
prototype
Name
Source
Animal
Human Arthropods
Sentinel Wild
A AN 8 Mucambo x + + + +
AN 7526 EEE + + +
H 407 Mayaro + + +
AR 10315 Aura x +
AR 13136 Una x +
AR 35645 Pixuna X + +
AN 70100 WEE +
B H 111 Yellow fever + + +
AN 4116 Bussuquara x + + +
H 7445 Ilhéus + + +
AR 23379 St. Louis + + +
C AN 17 Oriboca x + + + +
AN 974 Murutucú x + + + +
AN 15 Marituba x + +
AN 848 Apeú x + + + +
AN 3994 Caraparú x + + + +
AN 12751 Itaquí x + + + +
AN 10709 Nepuyo + + +
Guama H 151 Catú x + + + +
AN 277 Guamá x + + + +
AR 12590 (Mojú) x + + +
Capim AN 8582 (Capim) x + + +
AN 10615 (Guajará) x + + +
AN 20076 (Bush Bush?) + +
Mirim AN 7722 (Mirim) x + +
Bunyamwera H 12208 Guaroa +
AR 7272 (Maguari) x +
AR 8226 Kairi + + +
AR 32149 (Sororoca) x +
California AR 8033 Melao +
Wyeomyia AR 278 (Tucunduba) x +
AR 671 (Taiassui) x +
Simbú AN 19991 Oropouche + + +
AN 84785 (Utinga) x +
Turlock AN 32260 Turlock + +

Table 1 (continued)
Group
or
complex
Belém
prototype
Name
-59-
Human
Source
Animal
Sentinel Wild
Arthropods
Anopheles A AN 35112 (Lukuni) +
Phlebotomus
fever H 22511 (Candirú) x +
AN 24262 Icoaraci x +
AN 46852 (Anhangá) x +
AN 47693 (Bujarú) x +
AN 64582 Itaporanga + + +
Changuinola AN 28873 (Irituia) x +
Untyped +
Indiana VSV AR 39377 Cocal +
Timbó AN 41787 Timbó x +
AN 42217 Chaco x +
Cotia AN58058 (Cotia?) +
Tacaribe AN 70563 (Amaparí) + +
Ungrouped AN 73 Tacaiuma x + +
AN 24232 (Piry) x § +
AN 27326 (Pacui) x +
AN 27639 (Acará) x + +
AN 40290 Marco x +
AR 40578 (Jurona) x +
AR 50117 (Tembe) x +
AN 67949 (-) x +
Total 56 types 38 16 25 35 38
x Original isolation made in Belém.
§ Laboratory infection.
( )Unpublished.

Group or
complex
Belem
prototype
-60-
Table 2
Virus infections by type and year, 1954–1965
Name
Year
54 55 56 57 58 59 60 61 62 63 64 65 Total
A AN 8 Mucambo x 1 7 10 14 1 12 22 5 25 6 13 116
AN 7526 EEE 1 4 2 2 10 28 5 19 70
H 407 Mayaro 6 2 1 29 3 1 42
AR 10315 Aura x 1 1 1 2 1 6
AR 13136 Una x 2 2 7 1 12
AR 35645 Pixuna x 3 1 4
AN 70100 WEE 1 1
B H 111 Yellow fever 9 16 4 2 1 32
AN 4116 Bussuquara x 2 7 2 4 6 16 8 45
H 7445 Ilhéus 2 1 2 13 4 1 23
AR 23379 St.Louis 3 5 8
C AN 17 Oriboca x 1 8 7 3 11 15 11 5 5 11 8 85
AN 974 Murutucú x 5 4 1 9 10 5 12 1 12 7 66
AN 15 Marituba x 1 3 3 4 1 5 4 3 3 27
AN 848 Apeú x 5 5 7 2 4 2 2 2 3 3 35
AN 3994 Caraparú x 5 6 9 11 33 64 61 20 97 60 25 391
AN 12751 Itaquí x 1 2 13 38 26 6 41 38 17 182
AN 10709 Nepuyo 1 2 1 2 6
Untyped 4 1 5
Guamá H 151 Catú x 3 1 5 4 33 29 28 17 39 19 26 204
AN 277 Guamá x 6 10 2 8 24 41 51 17 49 52 47 307
AR 12590 (Mojú) x 2 9 22 13 10 33 39 12 140
Untyped 1 2 22 19 4 3 1 52
Capim AN 8582 (Capim) x 1 1 8 9 4 20 10 53
AN 10615 (Guajará) x 2 13 8 4 10 3 40
AN 20076 (Bush Bush?) 2 2 2 1 3 10
Mirim AN 7722 (Mirim) x 1 3 2 1 3 10
Bunyamwera H12208 Guaroa 4 1 5
AR7272 (Maguari) x 1 2 1 4
AR8226 Kairi 1 1 1 1 1 5
AR32149 (Sororoca) x 6 6
California AR 8033 Melao 2 1 3
Wyeomyia AR 278 (Tucunduba) x 2 1 2 1 2 8
AR 671 (Taiassui) x 2 1 1 1 5
Untyped 2 1 1 3 1 1 1 10
Simbu AN 19991 Oropouche 2 16 18
AN 84785 (Utinga) x 1 1

Table 2 (continued)
Year
54 55 56 57 58 59 60 61 62 63 64 65 Total
Name
Belem
prototype
Group or
complex
Turlock AN32260 Turlock 1 2 3
Anopheles A AR 35112 Lukuni 1 1
Phlebotomus
fever H 22511 (Candirú) x 1 1
AN 24262 Icoaraci x 1 1 7 1 10
AN 46852 (Anhangá) x 1 1
AN 47693 (Bujarú) x 1 1
AN 64582 Itaporanga 1 5 2 8
Untyped 1 1
Changuinola AN 28873 (Irituia) x 1 1
Untyped 1 2 1 4
Indiana VSV AR 39377 Cocal 1 1
Timbó AN 41787 Timbó x 5 1 6
AN 42217 Chaco x 2 2 4
Cotia AN58058 (Cotia?) 1 1
1 14 15
Tacaribe AN 70563 (Amaparí) x
Ungrouped AN 73 Tacaiuma x 1 1 2 1 5
AN 24232 Piry x 1 1
AN 27326 Pacui x 1 4 2 7
AN 27639 Acara x 2 1 3 2 8
AN 40290 Marco x 3 1 4
AR 40578 Jurona x 1 1
AR 50117 Tembe x 1 1 2
AN 67949 ( - ) x 1 1
Unidentified 3 1 4
T o t a l 56 types 38 12 70 57 48 66 181 291 355 139 377 310 228 2128
x Original isolation made in Belém.
( ) unpublished

-62 -
'l'able 3
V1ra888 isolated., b7 tJpe am source, 196>
S8D~el W11d

- 63 -
Table 4
Viruses from Sentinel Mice, 1965
Date Virus Site Trap Strain AN Group CM
Jan. 5 Catú IPEAN hood 80520 7464
9 Guajará IPEAN hood 81003 7475
10 Oriboca Utinga hood 80604 7478
( “ ( “ “ 80605 “ )
12 Itaqui Utinga hood 80870 7482§
( “ ( “ “ 80767 “ )
13 Caraparú IPEAN hood 80871 7483
16 Caraparú IPEAN hood 81013 7489§
17 Caraparú Utinga hood 81014 7492
19 Caraparú IPEAN hood 81167 7495
( “ ( “ “ 81250 “ §)
22 EEE IPEAN hood 81340 7501
23 Guamá Utinga hood 81374 7504
26 Itaqui Utinga hood 81435 7510§
( “ ( “ “ 81447 “ )
28 Guamá Utinga hood 81533 7514
Feb. 2 Guamá IPEAN blower 81921 7523
3 Oriboca IPEAN hood 81864 7527
3 Mojú Utinga hood 81724 7528
9 Guamá IPEAN blower 82091 7539
11 Catú Utinga hood 82075 7549
( “ ( “ “ 82093 “ )
( “ ( “ “ 82094 “ §)
12 Caraparú IPEAN hood 82095 7554§
( “ ( “ “ 82123 “ )
14 Guamá IPEAN hood 82138 7565
14 Guamá Utinga hood 82439 7566
16 Itaqui Utinga hood 82306 7577
24 EEE IPEAN hood 82635 7622
24 EEE IPEAN blower 82912 7624
25 Neuyo IPEAN blower 83008 7625
26 Mojú Utinga hood 83009 7632
26 Nepuyo IPEAN (Culex
spp.) §§ 83010 7634
Mar. 3 Caraparú Utinga hood 83011 7668
( “ ( “ “ 83012 “ §)
4 Mirim IPEAN (Culex(M)
taeniopus) 83378 7672
4 Mojú Utinga hood 83208 7675
5 Itaqui Utinga hood 83081 7680
6 Guamá Utinga hood 83277 7685
8 Guamá Utinga hood 83278 7692§

- 64 -
Table 4 – cont.
Date Virus Site Trap Strain AN Group CM
Mar. 11 EEE IPEAN hood 83379 7708
11 Mojú Utinga hood 83516 7709
( “ “ “ 83517 “ §)
12 Bush Bush IPEAN blower 83631 7717
13 Itaqui Utinga hood 83424 7732
14 Caraparú Utinga hood 83543 7737
16 Guamá IPEAN blower 83544 7745
17 EEE IPEAN hood 83545 7749
18 Oriboca IPEAN hood 83633 7755
18 Catú Utinga hood 83686 7756
23 Guamá IPEAN blower 83930 7782
24 Catú Utinga hood 83903 7788
26 Itaqui Utinga hood 83932 7798
26 Guamá Utinga hood 84008 “
28 Mojú Utinga hood 83928 7808
29 Caraparú IPEAN hood 83990 7812
29 Mojú Utinga hood 84094 7813
30 Catú IPEAN hood 84095 7814
30 Itaquí Utinga hood 84011 7815
31 Oriboca IPEAN hood 84190 7820§
31 Bush Bush IPEAN (Culex spp.)§§ 84269 7823
Apr. 2 Caraparú Utinga hood 84101 7836§
( “ “ “ 84100 “ )
3 Mojú IPEAN hood 84434 7840
4 Guamá Utinga hood 84279 7852
6 EEE IPEAN hood 84273 7860
6 Gr.Capim Utinga hood 84381 7861
7 Capim IPEAN (Culex spp.)§§ 84461 7868
8 Bussuquara Utinga hood 84436 7875
10 Caraparú Utinga hood 84456 7887
12 Itaquí Utinga hood 84536 7899
( “ “ “ 84650 “ §)
13 Caraparú IPEAN blower 84652 7906§
14 Guamá Utinga hood 84712 7908
17 Catú Utinga hood 84978 7933
19 Caraparú Utinga hood 85172 7944
20 Guamá IPEAN blower 84946 7945
20 Guamá Utinga hood 84947 7947
21 Capim IPEAN (Culex spp.)§§ 84948 7954
22 Caraparú IPEAN blower 85175 7956
22 Catú Utinga hood 84950 7958
2 3 Caraparú IPEAN blower 84952 7962
24 Guamá Utinga hood 85057 7971
25 Caraparú Utinga hood 84954 7977§
27 Guamá IPEAN (Culex B17)§§ 85058 7990
29 Catú Utinga hood 85180 7999
30 Guamá Utinga hood 85364 8014
( “ “ “ 85365 “ §)
30 Caraparú IPEAN blower 85181 8020

- 65 -
Table 4 – cont.
Date Virus Site Trap Strain AN Group CM
May 2 EEE Utinga hood 85367 8028
20 Mojú Utinga hood 86117 8104
22 Catú Utinga hood 86086 8110§
( “ “ “ 87287 “ )
23 Itaqui Utinga hood 86228 8112
( “ “ “ 86229 “ §)
24 Guamá Utinga hood 86151 8114§
28 Guamá IPEAN hood 86482 8121
June 4 Guamá Utinga hood 86723 8140
8 Guamá Utinga hood 86952 8155
10 Guamá IPEAN hood 87037 8160
11 Caraparú Utinga hood 86953 8164§
12 Caraparú IPEAN hood 86954 8169§
( “ “ “ 87293 “ )
12 Caraparú Utinga hood 87126 8170§
14 Catú Utinga hood 87182 8174
( “ “ “ 87295 “ )
15 Catú Utinga hood 88053 8177
( “ “ “ 88054 “ )
16 Itaquí Utinga hood 87290 8180§
17 Itaquí Utinga hood 87300 8182§
( “ “ “ 88095 “ )
18 Murucutú IPEAN hood 88056 8193
19 Murucutú IPEAN hood 87301 8195
19 Murucutú Utinga hood 88057 8196
21 Guamá Utinga hood 87758 8200
23 Guamá Utinga hood 87813 8206
24 Guamá IPEAN hood 87760 8207
29 Guamá IPEAN hood 87979 8224
Jul. 9 Murutucú IPEAN hood 89311 8255
10 Guamá IPEAN hood 88985 8265
13 Mucambo Utinga hood 88640 8272
16 Itaquí IPEAN blower 88800 8279
18 Catú Utinga hood 89157 8289
21 Itaquí IPEAN hood 89352 8296
26 Oriboca IPEAN hood 89420 8314
Aug. 1 Mirim IPEAN hood 90149 8334
5 Guamá IPEAN blower 90116 8345§
11 Mojú IPEAN hood 90961 8368
14 Itaquí IPEAN hood 90545 8380
( “ “ “ 90666 “ §)
17 Catú IPEAN hood 91035 8386
( “ “ “ 94216 “ )
25 Apeú IPEAN hood 92566 8415
25 Itaquí IPEAN blower 92109 8417
27 Guamá IPEAN hood 94217 8422

- 66 -
Table 4 – cont.
Date Virus Site Trap Strain AN Group CM
Sep. 26 Guamá IPEAN hood AN 93484 8519
28 Catú IPEAN blower 94284 8524
30 Oriboca IPEAN blower 93520 8535
( “ “ “ 93521 “ §)
Oct. 7 Guamá IPEAN hood 93837 8556
8 Catú IPEAN blower 94510 8561
17 EEE IPEAN hood 94175 8596
( “ “ “ 94201 “ §)
29 Catú IPEAN blower 94558 8629
Nov. 1 Oriboca IPEAN hood 94559 8638§
3 Guamá IPEAN hood 94755 8642
15 Mucambo Utinga hood 95067 8682
Dec. 12 Guamá IPEAN blower 95733 8742
21 Caraparú IPEAN blower 95892 8778
30 EEE IPEAN hood 96746 8812
30 EEE Utinga hood 96747 8813
§ isolated from mother mouse
§§ exposed to bites of wild-caught mosquitoes of this species
in the laboratory

- 67 -
Table 4a
Viruses from Sentinel Monkeys and Chickens, 1965
Date Virus Site Sentinel Strain
Mar. 15 Mucambo Utinga MY 393 Cebus AN 83402
26 Mucambo Utinga MY 390 Cebus AN 83866
Oct. 15 Caraparú Utinga MY 401 Cebus AN 94112
25 Mucambo Utinga MY 401 Cebus AN 94396
Dec. 16 EEE IPEAN PT 181 chicken AN 95810

- 68 -
Table 5
Viruses isolated from Wild Vertebrates, 1965
Date Virus Strain Vertebrate Source Site
Jan. 7 Guamá AN 80395 RO 6273 Nectomys blood Utinga
18 Amaparí AN 81087 RO 6323 Oryzomys goeldi viscera Amapá
18§ Amaparí AN 81088 RO 6324 Oryzomys goeldi viscera Amapá
18 Amaparí AN 81092 RO 6329 Oryzomys goeldi viscera Amapá
22 Murutucú AN 81261 MA 1719 Marmosa sp. blood Utinga
Feb. 11 Catú AN 82045 RO 6369 Oryzomys goeldi blood Utinga
23 Catú AN 82539 RO 6386 Proechimys blood Utinga
26 Guamá AN 82765 RO 6362 Nectomys blood Utinga
Mar. 5 Mojú AN 83058 RO 6396 Proechimys blood Utinga
17 Guamá AN 83454 RO 6368 Oryzomys goeldi blood Utinga
19 Guamá AN 83607 “ “ “ “ “
19 Caraparú AN 83609 RO 6408 Oryzomys goeldi blood Utinga
24 Bussuquara AN 83746 RO 5762 Proechimys blood Utinga
26 Guamá AN 83856 MA 1768 Didelphis blood Utinga
Apr. 2 Catú AN 84022 RO 6439 Oryzomys goeldi blood Utinga
22 Bussuquara AN 84778 RO 6395 Proechimys §§ blood Utinga
22 Gp.Simbu AN 84785 ED 153 Bradypus blood Utinga
tridactylus
( “ “ AN 84773 “ “ viscera “ )
23 Guamá AN 84857 RO 6387 Proechimys blood Utinga
May 11 Guamá AN 85607 RO 6395 Proechimys§§ blood Utinga
18 Mojú AN 85959 RO 6401 Nectomys blood Utinga
24 Murutucú AN 86016 RO 6149 Proechimys blood Utinga
24 Itaqui AN 86026 RO 6536 Nectomys blood Utinga
June 4 Apeú AN 86650 MA 1846 Calluromys blood Utinga
7 Amaparí AN 86901 RO 6562 Oryzomys goeldi viscera Amapá
16 Guamá AN 87171 RO 6516 Proechimys blood Utinga
25 Catú AN 87774 RO 6612 Oryzomys blood Utinga
Jul. 12 Amaparí AN 88485 RO 6637 Oryzomys
macconnelli viscera Amapá
21 Mucambo AN 88995 199775 Pipra
erythrocephala blood IPEAN
21 Amaparí AN 89033 RO 6623 Oryzomys goeldi viscera Amapá

- 69 -
Table 5 – cont.
Date Virus Strain Vertebrate Source Site
Aug.
5 Amaparí AN 90026 RO 6699 Neacomys viscera Amapá
26 Amaparí AN 92098 RO 6744 Neacomys viscera Amapá
26 Amaparí AN 92099 RO 6745 Neacomys viscera Amapá
30 Amaparí AN 92576 RO 6764 Neacomys blood Amapá
Sep. 3 Amaparí AN 92822 RO 6764 Neacomys viscera Amapá
9 Guamá AN 92909 MA 2008 Metachirus blood Utinga
20 Amaparí AN 93159 RO 6811 Oryzomys goeldi viscera Amapá
22 Amaparí AN 93233 RO 6832 Neacomys viscera Amapá
Oct. 6 Murutucu AN 93731 RO 6548 Proechimys blood Utinga
15 Moju AN 94021 RO 6558 Proechimys blood Utinga
15 Catú AN 94103 CH 2074 bat sal.glands Amapá
Nov. 3 Mucambo AN 94768 RO 6907 Oryzomys goeldi haemorrhagic
urin. Utinga
12 Icoaraci AN 94945 RO 6854 Proechimys blood Utinga
Dec. 15 Caraparú AN 95793 RO 6956 Proechimys blood Utinga
16 Mucambo AN 95820 RO 6934 Oryzomys goeldi blood Utinga
22 Mucambo AN 95889 RO 6993 Nectomys blood Utinga
§§ same individual
§ date of sacrifice: date of capture = 23 December 1964.

- 70 -
Table 6
Viruses isolated from Arthropods, 1965 captures
Date Pool Nº Virus Arthropod Site Bait Method
AR
Jan. 1-15 81105 Guajara Culex B9 IPEAN mouse hood
4-14 81144 Catu Culex B9 IPEAN human hand
4-14 81176 Caraparú Culex B9 IPEAN human hand
16-31 81591 Capim Culex B1 Utinga mouse hood
16-31 81594 Mucambo Culex B9 Utinga mouse hood
16-31 81596 Mucambo Culex B19 Utinga mouse hood
20-28 81651 Caraparú Culex (C.)
coronator IPEAN human hand
16-31 81828 EEE Culex (M.)
taeniopus IPEAN chick protected
16-31 81869 Capim Culex B1 IPEAN mouse hood
81926 Amapari Gamasidae Amapá Oryzomys combed
Feb. 1-15 82507 EEE Culex spp.
(fed) IPEAN mouse hood
1-15 82520 Capim Culex B1
(unfed) IPEAN mouse hood
1-15 82903 Wyeomyia Wyeomyia spp. Utinga human hand
16-28 83412 Bush Bush Culex spp.
(unfed) Utinga mouse hood
Mar. 18-30 84249 Catu Culex B9 IPEAN human hand
16-30 84577 Bussuquara Culex B1 Utinga human hand
16-30 84600 Bussuquara Mansonia
titillans Utinga human hand
29-31 84685 Guama Culex B9 Utinga none suction
16-31 84719 Mucambo Culex B7
(fed) IPEAN mouse hood
Apr. 2 84049 Capim Culex spp. IPEAN mouse blower
2 84050 Guajara Culex spp. IPEAN mouse blower
14 84538 Capim Culex spp. IPEAN mouse blower
1-14 84889 EEE Culex(M.)spp. Utinga none suction
1-13 84992 Catu Culex B9 IPEAN human hand
1-13 84994 Guama Culex B19 IPEAN human hand
30 85094 Apeu Culex B19 IPEAN mouse blower
1-15 85330 Capim Culex B1
(unfed) IPEAN mouse hood
1-15 85344 EEE Culex B1
(fed) Utinga mouse hood
2-20 85641 Tacaiuma Haemagogus
spp. Amapá human hand
16-30 86152 Capim Culex spp.
(fed) IPEAN mouse hood
16-30 86176 EEE Culex B8
(fed) Utinga mouse hood

- 71 -
Table 6 – cont.
Date Pool Nº AR Virus Arthropod Site Bait Method
May 4-13 86260 Bussuquara Mansonia
venezuelensis IPEAN human hand
June 4 86621 Catu Culex B9 IPEAN mouse blower
4 86628 Guama Culex (M.)
taeniopus IPEAN mouse blower
1-15 87476 Itaporanga Culex spp. IPEAN human hand
25 87737 EEE Culex (M.)
taeniopus IPEAN mouse blower
18-30 88291 Itaporanga Mansonia
venezuelensis IPEAN human hand
16-30 89475 Bussuquara Culex spp.
(unfed) IPEAN mouse hood
1-30 89546 Mirim Aedes serratus
(fed) Utinga mouse hood
Aug. 4 90010 Bussuquara Culex spp. IPEAN mouse blower
6 90095 Itaqui Culex B7 IPEAN mouse blower
6 90098 Guama Culex B9 IPEAN mouse blower
11 90282 Guama Culex B9 IPEAN mouse blower
19-30 92965 Mucambo Mansonia spp. IPEAN human hand
Sep. 1-9 93375 Aurá Aedes serratus IPEAN human hand
21-30 94240 Oriboca Culex B9 IPEAN mouse blower
Oct. 29 95078 Catu Culex B9 IPEAN mouse blower
Dec. 21-2 96240 EEE Culex (M.)
taeniopus IPEAN none suction
16-31 96304 EEE Culex (M.)
taeniopus tamarin mouse blower

- 72 -
Table 7
Viruses isolated from Arthropods, 1964 captures, identified 1965.
Month Pool Nº Virus Arthropod Site Bait Method
August AR 79437 Mucambo Mansonia venezuelensis Utinga human hand
October AR 79981 Wyeomyia Mansonia arribalzagai Utinga human hand
complex
December AR 80353 Guama Mansonia venezuelensis Utinga human day
AR 80468 Itaqui Culex B9 Utinga mouse hood
AR 80475 Itaqui Culex B7 IPEAN mouse hood
AR 80479 Itaqui Culex (M). spissipes IPEAN mouse hood
AR 80652 Caraparu Culex B7 IPEAN human night
AR 80653 Catu Culex B9 IPEAN human night
AR 80663 EEE Culex (M) taeniopus IPEAN human night
AR 80740 Oriboca Culex B9 IPEAN human night

Mosquito
- 73 -
Table 8
Species of mosquito giving virus
by inoculation or laboratory transmission, 1965
Nº
tested
Nº
isolated
Rate Virus
CULICINI
Aedes serratus 4,543 2 2,300 Aurá, Mirim
Culex spp. 12,428 13 1,000 Bush Bush, Bussuquara, Capim,
EEE,
Guajará, Itaporanga, Nepuyo
B1 571 6 100 Bussuquara, Capim, EEE
B7 2,680 2 1,000 Itaqui, Mucambo
B8 362 1 EEE
B9 12,009 12 Caraparu, Catú, Guajará, Guamá,
Mucambo, Oriboca
B17 5,593 1 Guamá
B19 2,802 3 900 Apeú, Guamá, Mucambo
(Culex)coronator? 519 1 Caraparú
(Melanoconion)spp. 655 1 EEE
(M)taeniopus 11,279 6 2000 EEE, Guamá, Mirim
Haemagogus spp. 925 1 Tacaiuma
Mansonia spp. 6,177 1 Mucambo
titillans 226 1 Bussuquara
venezuelensis 16,033 2 8,000 Bussuquara, Itaporanga
SABETHINI
Wyeomyia spp. 4,168 1 Wyeomyia
Total: 16 species 80,970 54

- 74 -
Table 9
Rainfall (mm.)
Utinga IPEAN Serra do Navio
1965 1965 a 1964 b 10-yr.Avg. 1965 1964 1957-62 Avg.
January 335 685 335 222 339 249
February 338 670 402 226 280 224
March 449 623 422 354 244 284
April 411 284 409 131 207 350
May 426 415 256 396 245 322
June 195 129 204 119 316 214
July 95 41 149 110 186 224
August 70 62 112 180 189 112
September 108 243 110 84 73 67
October 169 172 (81)c 107 43 69 76
November 88 132 (30)d 128 57 54 84
December 215 190 (240)d 224 51 72 154
Total 2899 - (3503) 2858 1973 2274 2360
a = data from new station in experimental area
b = data from old station near Administrative building, closed Oct.1964
c = data from US Consulate, Belem
d = data from Utinga station

- 75 -
Table 10
Vertebrate specimens examined for virus isolation or serology, 1965
Total
Species
Total
Order
Total
Class
MAMMALIA 8,085
PRIMATA 1,250
Homo sapiens 1,071
Cebus paella 85
Saimiri sciureus 23
Tamarin tamarin 58
Others 13
MARSUPIALIA 855
Calluromys sp. 246
Didelphis marsupialis 237
Marmosa spp. 148
Philander opossum 136
Metachirus nudicaudatus 48
Monodidelphis spp. 40
CHIROPTERA 2,570
EDENTATA 37
Bradypus tridactylus 28
Others 9
RODENTIA 3,075
Nectomys aquaticus 187
Oecomys concolor 3
Oryzomys spp. 927
Neacomys guianae 226
Proechimys guyanensis 1,702
Others 30
CARNIVORA 10
Canis familiaris 3
Felis domestica 3
Others 4
ARTIODACTYIA 288
Bos indicus 255
Equinus 57
Caprinus 5
Ovis 1

4"
o=t.1Du8d
.t~ -
REPrII.IA!.
~ ,..
~UAMA.!&.
n~~
-76-
Total
Spec1es
Theo8d8)~ ra~! ~~'~!8 1.
IGtWfID&X
19uana. iguana
~!Jr\1.I ~&1a18
'lro~ 8 t,caoqUA 'b18
Ur81O8codon SUpsrC:W.~.
2
27
228 2
scmc:ma,
abOQ'a abOIva 1
'1~:II"a
.&81 va aS. va
~~ lemnU~~
'l\1!Y'D8mb1a nigropmcta'bJ8
AMlmBIA.
-~-~-
NO'l' ro}]rrIFIm
125 92
BOIDAB 3
&1to 8pp. .3S
6
'l'ot8i1.
arder
'l'otaQ.
Claaa
Tota.1. 10;733
JS

- 77 -
Table 11
Changes in HI antibody in birds (negative = < 1:10)
Species Group A Group B Other groups
Chloroceryle inda
§ 654-038 neg UNA 1:20
Pyriglena
653-028 neg UNA 1:20
Saltator
654-072 neg EEE 1:20
Saltator maximus
654-096 EEE 1:40 neg
Thamnophilus aethiops
653-101 EEE 1:20 neg
Phlegopsis
654-048 WEE 1:10 1:40
Galbula albirostris
654-053 neg WEE 1:20 neg SLE 1:40
Thamnophilus aethiops
653-130 WEE 1:10 –neg- 1:20 SLE 1:20 Neg 1:40 ITA 1:20 neg 1:20
neg JUR 1:40
Thamnomanes neg ILH 1:40
71-82554 neg SLE 1:40
Phlegopsis neg ILH 1:40
654-571 neg SLE 1:320
Automolus infuscatus
654-006 ITA 1:10
1:40 1:10
Saltator maximus
654-091 GUAROA 1:40 neg
Glyphorhynchus
71-82520 neg JUR 1:20
Ramphocelus
653-008 neg JUR 1:20
Hylophyldx
653-014 neg TAC 1:20
§ ring number

(11)
Fie1d N g
--~--
198875
198879
198898
198931
199128
199198
199206
199258
199378
199383
199550
199678
199743
199800
199976
31517
31576
31686
317«,
31718
31896
32017
33).J.31
33463
33464
33510
653-01)-:. (rin~ ng)
§ canopy bird
§ § ground bird
Total 27
( 4)
-78
Tab1e 12
Itaporanga RI Antibody in Rirds
Titre
-.,~ ~~i.es
1:20
1:20
1:20
1:20
1:20
1:20
1:40
1:40
1:80
1:20
1:40
1:40
1:80
1:80
1:20
1:40
1:80
1:80
1:20
1:40
1:20
1:20
1:40
1:20
1:20
1:20
1:40
§ Rhamphastos cuvieri
§§ Hypocnemoides
§ Thamnomanes caesius
§ Thamnophilus aethiops
§ Thamnomanes caesius
§ § l"tvnnotherula hauxwelli
§ § Hypocnemoides
§ Thamnophilus aethiops
§ Attila spadiceus
§ Thamnomanes caesius
§ Cercomacra cenerascens
§§ Automolus infuscatus
§ ThaJnnophilus aethiops
§ Thamnomanes
Turdus fumigatus
§ ThaJnnomanes
Unidentified
§ Accipitor bicolor
§ Thamnophilus aethiops
§ Thamnomanes caesius
§ ThaIIn'lornanes
Cora~
§ ThaJnnophilus aethiops
§ Thamnophilus aethiops
Unidentified
Cora~
Itrlophylax

Table 13
Ectoparasites inoculated from IPEAN and Utinga forests
Other
acariens Pulicidae Dermatobium Other Total
Host Ixodidae Gamasidae
Oryzomys 71(11) § 8533(53) 11(3) 125(12) 29(22) - 8769(101)
Nectomys 6(2) 915(12) - 3(2) 29(16) - 953(32)
Proechimys - - - 5(1) 18(16) 100(1) 123(18)
Oecomys 2(1) 16(2) - - - - 18(3)
Rhipidomys - 97(2) - - - - 97(2)
Marsupialia 94(15) 4(1) - - 2(2) - 100(18)
Chiroptera 1(1) - - - - 24(4) 25(5)
Edentata 19(7) - - - - - 19(7)
Aves 47(24) - - - - 2(2) 49(26)
Reptilia 21(1) - - - - - 21(1)
Total 261(62) 9565(70) 11(3) 133(15) 78(56) 126(7) 10,174(213)
§ ( ) = n° of pools. No isolations were made.

Animals
- 80 -
Table 14
Animals captured and specimens collected
At Serra do Navio, Amapá, 1965.
N° of
Material inoculated and preservation
Glycerine - 60° C
Brown
Captures Recapt Viscera Sal.Cl. Viscera Sal.Cl. fat Urine Serol.
Blood
Virus +
serol. Inoc.
Aves 224 - - - - - - - 224 - -
Chiroptera 472 - 150 137 190 204 27 - 300 48 2
Didelphis 78 8 56 - - - - - 72 4 -
Metachirops 86 4 63 - - - - - 78 1 -
Marmosa 10 - 5 - 2 - - - 4 3 -
Monodelphis 15 - 7 - 9 - - 1 3 9 -
Metachirus 16 - 14 - - - - - 14 - -
Calluromys 3 - 3 - - - - - 3 - -
Proechimys 108 1 12 - 79 - - 53 14 78 -
Oryzomys
goeldi
O.maccon-
nelli
Neacomys
guianae
97 - 4 - 91 - - 3 2 90 -
13 - 1 - 12 - - 4 - 12 -
124 - 18 - 99 - - 10 5 93 6
Saguinus 10 - 10 - - - - - 9 1 -
Other § 11 - 9 - - - - - 6 - -
Total 1,267 13 352 137 482 204 27 71 734 339 8
§ includes 3 Myoprocta, 2 Tamandua, 3 Sciuridae, 1 Echimys, 1 Tupinambis and 1 turtle.

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Table 16
Ectoparasites inoculated from Amapá
Other
acarines Pulicidae Dermatobium Other Total
Host Ixodidae Gamasidae
Neacomys - 638(9)§ - - - - 638(9)
Oryzomys 8(3) 1612(16) - 10(5) 7(6) - 1637(30)
Proechimys 19(1) 80(4) 25(2) 8(2) 1(1) - 133(10)
Myoprocta 1(1) - - - - - 1(1)
Marsupialia 285(21) 24(3) - 1(1) - - 310(25)
Chiroptera - - - - - 7(1) 7(1)
Edentata 59(5) - - - - - 59(5)
Equine 49(6) - - - - - 49(6)
Reptilia 14(1) - - - - - 14(1)
Aves 1(1) - - - - - 1(1)
Total 436(39) 2354(32) 25(2) 19(8) 8(7) 7(1) 2849(89)
§ ( ) = N° of pools. Only one isolation was made (Amaparí virus from a pool of Gamasidae
off Oryzomys.)

Tabl. 14
~rtc Platon
Sagu1nU8
Proe~
Brad1P18
l'alodelP1i8
Scu1ru8
Aves
Sen'& do Navio
~Pro~~
0I7J.01T0"8
Neac.-
D1delJi1ü
~~a: Metachirops
~tac~
~.
lblodelIi1i8
CallurOJl'B
~FOCta
Tmf8ndua
~opter&
Aves
Total
_..,-
-83 -
3.1
BI test r88ul ta m ani--1 sera .troa ,,-pá, 1965
uroup
A
- -
-.
- -
4- o
-
-
-
-
- 9
üroup
B
-
-
-
-
-
1 -
1 ------ -74
uroup
C ~
-
1 ---
-
-
1 -
1 ----- ---
urou~
~
-
1 ---
-
4
2 -
- -
-
- -
SS5
uroup
HüebotoJ!!U8
--, --,
2
3-
-
-
16
D .
1
4
S-~ - -
-§§
3
'.L'O~ S
test8d
S 8
1 1
1
4
88
lCJl
89
69
7l
16
.3
9
.3
2
1
304
215
13 .3 7 .34 99l
§ Smne ot these lera ~ also teeted 11'1. th B\m)'8Dfera group and Capim antigens,
wi~ negati" resulte. 007 one eeru8 IpeC~WU c~ted trom .ach anial;
ml7 titree or 1:20 or greater aga1n8t 8 uniu ot ant1g8'l are coneidered
pos i tJ. ,'c .
§§ ~ Taca1UJl& ~it1V8.
§§§ CZ1ly 52 t88ted~ alI negative.

Estimated age
-84-
Table 18
HI results on sera of Kayapo-Gorotire Indians
Mayaro
Total
Group A
Total
Group B
Yellow
fever Ilheus
Indef.
Group B
Total
tested
M F M F M F M F M F M F M F
0-4 0 3 0 3 0 0 0 0 0 0 0 0 3 4
5-9 4 4 4 4 0 1 0 0 0 1 0 0 14 17
10-14 10 5 11 5 2 2 0 0 1 2 1 0 22 12
15-19 4 1 5 1 3 2 1 0 1 2 1 0 10 10
20-24 3 4 3 4 4 2 0 0 2 2 2 0 9 16
25-29 10 4 11 4 7 4 0 0 3 2 4 2 18 12
30-34 3 3 3 3 5 8 0 3 4 4 1 1 10 13
35-39 2 5 3 5 4 2 2 0 0 2 2 0 11 7
40-44 5 5 5 8 4 3 1 0 0 2 3 1 8 10
45-49 1 1 2 1 2 1 0 0 2 1 0 0 3 2
50 + 2 1 2 1 1 1 1 0 0 1 0 0 4 4
Subtotal 44 36 49 39 32 26 5 3 13 19 14 4 112 107
Total 80 88 58 8 32 18 219
% 36 40 27 4 15 8

Table 19
Results of CF tests using pooled antigens
Immune
Titre Immune
Titre
ascetic fluids Homologous Pool ascetic fluids Homologous Pool
GROUP A N° 1 GROUP 5 N° 5
Mucambo (AN8) 4i. 32/256 § 32/64 Guajará (AN 10615) 3i. 64/64 64/64
EEE (AN 7526) 4i. 32/256 32/256 Capim (AN 8582) 3i. 128/256+ 128/256
Mayaro (H407) 5i. 4/16 4/16 Bush Bush (AN 20076) 4i. 128/256+ 128/64
“ (AR 20290) 5i. 4/16 4/16 Mirim (AN 7722) 128/256+ 128/64
Aurá (AR 10315) 4i. 64/64 32/64
Una (AR 13136) 4i. 32/64 16/16 GROUP 6 N° 6
Pixuna (AR 35645) 4i. 32/16 8/16 Oropouche (AN 19991) 3i. 128/256+ 128/256
WEE (AN 70100) 5i. 128/64 128/64 Turlock (AN 32260) 128/256 128/256
Lukuni (AR 35112) 5i. 16/256+ 16/256+
GROUP B N° 2 Irituia (AN 28873) 3i. 16/16 32/16
Bussuquara (AN 4116) 4i. 128/1024+ 64/1024 Cocal (AR 39377) 4i. 128/64 128/64
Ilhéus (H 7445) 2i 32/1024+ 32/1024+ Timbó (AN 41787) 3i. 32/256 32/64
SLE (AR 23379) 4i. 8/1024+ 8/1024+ Chaco (AN 42217) 4i. 64/256+ 32/256+
Yellow fever (H 111) 3i. 32/1024+ 32/1024
GROUP 7 N° 7
GROUP C N° 3 Icoaraci (AN 24262) 6i. 128/256+ 128/256+
Marituba (AN 15) 2i. 4/128 4/512+ Candirú (H 22511) 5i. 128/16 128/16
Oriboca (AN 17) 2i. 8/128 8/512+ Itaporanga (prototype) 2i. 4/4 4/4
Apeú (AN 848) 1i. 4/128 4/512+ Anhanga (AN 46852) 4i. 128/256+ 128/64
Murutucú (AN 974) 2i. 4/128 4/128 Bujarú (AN 47693) 32/16 32/16
Caraparú (AN 3994) 2i. 4/128 4/32
Itaquí (AN 12797) 5i. 4/32 4/128 GROUP 8 N° 8
Tacaiuma (AN 73) 4i. 16/32 16/32
GROUP BUNYAMWERA N° 4 Piry (AN 24232) 5i. 8/32 8/16
Guaroa (H 12208) 5i. 64/256 128/256+ Pacui (AN 27326) 5i. 32/512+ 32/64
Kairi (AR 8226) 128/64 128/256 Acará (AN 27639) 5i. 64/16 32/16
Maguarí (AR 7272) 5i. 128+/256+ 128+/256+ Marco (AN40290) 6i. 128/16 128/16
Sororoca (AR 32149) 4i. 128/256+ 128/256+ Jurona (AR 40578) 4i. 16/128 16/64
Tucunduba (AR 278) 2i. 128/256+ 128/256+ Tembe (AR 50117) 6i. 8/4 4/4
Taiassui (AR 671) 128/256+ 128/256+
Melao (AR 8033) 3i. 4/4 4/64
§ Antiserum titre / antigen titer.
Each pool contained all the antigens in the respective group.

Serum or immune
-86-
Table 20
Plaque inhibition test results with Group A viruses
N°
inj.
(zone diam. in mm.)
EEE (1)§
AN 7526
Mucambo (1)
AN8
Mayaro (1)
H407
Aura (3)
AR 10315
WEE (3)
AN 70100
EEE (AN 7526) 4 13.7 0 0 0 0
“ “ 2 8.0 - - - -
Mucambo (AN 8) 4 0 12.8 0 0 0
“ (AN10967) 1 0 14.5 0 0 0
Mayaro (H407) 5 0 0 18.8 0 0
“ (AR20290) 5 - 0 27.2 0 0
Aurá (AR 10315) 4 0 0 0 18.1 0
Una (AR 13136) 4 0 0 0 0 0
Pixuna (AR 35645) 4 0 0 0 0 0
WEE (AN 70100) 5 - 0 0 12.3 14.4
“ “ 3 - 0 0 12.0 18.8
“ “ 1 0 - - - -
§ ( ) nº of tests run. All tests run in duplicate with 2,400-12,000 pfu
in 100 mm Petri dishes, read after 3-4 days.

Table 27
Ground level mosquito captures by day on human bait, Utinga forest, 1965
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec
CULICINI
Aedes arborealis - 2 26 38 4 8 - - - - - - 78
argyrothorax - 7 22 9 15 14 3 - 2 1 4 1 78
fulvithorax - 31 21 21 10 24 10 3 3 7 6 7 143
fluvus - - - 1 - - - - - 1 - - 2
hortator § - - 1 - - - - 3 3 2 4 3 16
leucocelaenus § - - - - - 1 - - - - - - 1
oligopistus - - - - - - - - - - 2 - 2
scapularis - 120 100 72 10 39 13 24 41 555 99 402 1475
septemstriatus - 19 19 10 7 10 3 7 2 1 6 4 88
serratus - 185 155 101 37 102 214 323 709 740 249 239 3054
taeniorhynchus - 31 5 - - - - - - 1 - - 37
Culex spp. - 45 38 5 42 80 47 33 8 23 28 27 376
B 1 - 15 10 14 - - - - - - - - 39
B 7 - 1 1 - 1 1 1 - - - 1 - 6
B 8 - 3 3 1 - - - - 1 - - 2 10
B 9 - 5 6 2 4 10 3 5 1 1 6 - 43
B 17 - 3 1 - 3 - - - - - - - 7
B 19 - 10 4 2 7 16 10 13 4 3 2 - 71
(Carrollia) spp. § - - - - - 1 - - - - - - 1
(Culex) coronator? - 12 4 1 1 - 2 5 7 1 6 2 41
declarator - 2 1 - - - - - 1 1 - 1 6
pipiens - - - - - - - - - - - 4 4
(Melanoconion) spp. - 4 6 5 - - - - - - - - 15
caudelli? - - - - - 1 - - - - 4 - 5
spissipes - 4 4 1 - - - - - - - - 9
taeniopus - 1 1 1 - 1 - - - - - - 4
(Microculex) stonei § - - 10 3 - - - - - - - - 13
Haemagogus spp. - 10 17 - 4 8 7 - 1 21 14 5 87
Mansonia spp. - 596 608 456 338 588 358 425 149 127 124 71 3840
albicosta - 23 62 43 56 95 43 26 6 4 4 5 367
arribalzagai - 436 417 191 50 85 88 103 368 530 456 63 2787
titillans - 3 3 1 - 8 7 13 24 13 21 18 111
venezuelensis - 1113 1359 1376 902 1641 849 824 318 289 374 90 9138

Table 27 (continued)
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec
Orthopodomyia spp. § - - - - - - - - - 1 - - 1
Psorophora spp. - - - - - - - - - 1 - - 1
albipes - 89 - 29 3 15 2 2 3 25 6 8 182
cingulata - - - - 1 - - - 1 - - 1 3
ferox - 102 97 25 11 30 24 77 131 31 42 11 581
lutzi - 8 6 3 - 1 2 1 2 - 24 - 47
SABETHINI
Limatus spp. - - 7 - - 1 - - - - - - 8
durhami § - - 1 - - - - - - - 7 7 15
flavisetosus - 110 127 36 24 9 4 5 15 15 60 97 502
paraensis - 19 8 2 6 9 10 1 9 10 77 74 225
Phoniomyia spp. - 23 83 42 16 11 9 9 12 7 20 21 253
Sabethes (Sabethes) spp. § - - - - - - - - - - 1 - 1
belisarioi § - - - - - - 4 - - - - - 4
cyaneus § - - - - - - 6 6 3 2 3 1 21
(Sabethoides) chloropterus - - - - - - 1 - 1 - - - 2
Trichoprosopon digitatum - 6 17 85 91 25 5 6 10 6 6 11 268
edwardsianus - - 4 1 - - - - 2 - 2 2 11
magnus - 11 22 6 3 11 29 36 21 8 14 15 176
Wyeomyia spp. - 273 213 148 94 53 68 62 122 93 361 245 1732
(Dendromyia) aporonoma - 140 153 87 81 81 57 45 69 56 77 47 893
melanocephala § - 1 - - - - - - - - 21 36 58
ANOPHELINI
Anopheles (A.) eiseni § - - 1 - - - - - - - - - 1
(A.)intermedius - 2 2 - 2 1 5 10 12 1 3 - 38
(A.)mediopunctatus - - 2 1 - 1 - - - - - 1 5
(N.)oswaldoi - 12 32 7 3 2 6 13 20 1 1 2 99
(N.)triannulatus - 1 9 17 14 14 15 32 74 13 10 7 206
(S.)nimbus - 1 5 3 2 10 7 5 2 - - - 35
T o t a l - 3482 3693 2846 1842 3008 1912 2117 2157 2591 2145 1530 27322
Capture time (hours) - 31:30 23:00 17:30 19:00 24:00 26:30 30:00 30:00 23:30 24:30 25:40 275:20
Rate per hour - 111 160 163 97 125 72 71 72 110 87 59 99
§ these 13 spp. not taken in night catches on human bait at IPEAN.

Table 22
Ground level mosquito captures at night on human bait, IPEAN varzea, 1965
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec
CULICINI
Aedes arborealis - - 2 2 1 1 - - - - - - 6
argyrothorax 1 1 - 2 1 - - - - - 1 - 6
fulvithorax 1 - 6 - 1 2 1 2 4 1 - 1 19
fluvus 2 - - - - - 4 1 - 7 - - 14
scapularis 19 16 20 9 6 4 14 14 113 225 65 295 800
septemstriatus 12 - 6 1 - 1 10 7 1 1 2 2 43
serratus 57 7 2 - 1 10 77 24 59 219 27 19 502
taeniorhynchus - 3 - - - - - - - - - - 3
Culex spp. 11 18 14 1 9 106 10 50 24 2 39 10 294
B 1 7 - - 3 2 - - - - - - - 12
B 7 7 11 5 9 2 1 5 3 5 - 4 15 67
B 8 - - - - - - - - 1 - 1 9 11
B 9 115 59 27 53 47 37 12 23 38 13 37 33 494
B 17 3 - 29 20 13 20 1 11 2 2 1 4 106
B 19 70 30 23 44 38 60 53 38 114 118 130 120 847
(Culex) coronator? 9 2 1 - 1 - 2 - 6 7 23 16 67
declarator 2 - 1 - - - - - - - 4 5 12
(Melanoconion) spp. 4 2 1 19 2 1 - - - - - - 29
spissipes 64 19 3 9 3 7 3 - 7 2 2 2 121
taeniopus 45 35 28 39 78 67 6 7 28 112 58 24 527
Haemagogus spp. 1 1 - - - - - - - - - - 2
Mansonia spp. 47 172 114 144 107 123 149 198 58 - 21 5 1138
albicosta 1 3 - 1 - - 2 1 - - - - 8
arribalzagai 103 45 39 38 21 21 68 16 113 38 21 6 529
titillans 2 - - 4 2 - 10 3 20 - 1 2 44
venezuelensis 177 355 250 591 555 403 306 473 256 6 41 1 3414
Psorophora spp. 37 19 - - - - - 1 - - - - 57
cingulata - - - - - 1 - - 2 - - 2 5
ferox 7 2 3 - - 2 4 5 13 2 - 1 39
lutzi - - - - - - 1 - 1 - - - 2

Table 22 (continued)
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec
SABETHINI
Limatus flavisetosus 2 3 - - - 1 - - - - - - 6
paraensis - 1 - 1 - - - - - - - - 2
Phoniomyia spp. 1 1 - 1 - 1 - - - - - - 4
Trichoprosopon digitatum 4 5 14 23 17 20 14 8 7 2 3 3 120
magnus - - - - - - 1 - - - - - 1
Wyeomyia spp. 1 1 1 - - 3 - 2 - - - - 8
(Dendromyia) aporonoma - 1 1 - - - 2 1 - - - - 5
ANOPHELINI
Anopheles (A.) intermedius 4 - 1 1 2 1 - - 2 1 - - 12
(A.)mediopunctatus - - 3 5 1 1 6 3 17 - - - 36
(N.)oswaldoi 2 - 1 - - - - 1 7 - 2 2 15
(N.)triannulatus - - - 2 - - 1 1 5 - - - 9
(S.)nimbus 1 - 3 4 10 25 2 12 1 - - - 58
T o t a l 819 812 598 1026 920 919 764 905 904 758 492 577 9494
Capture time (hours) 15:00 10:25 8:20 12:00 8:00 10:00 8:20 8:00 10:00 8:00 8:00 9:00 114:45
Rate per hour 55 78 72 86 115 92 96 101 90 65 62 64 83

Table 23
Ground level mosquito captures at night on human bait, IPEAN capoeirão, 1965
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Total Varzea
total
CULICINI
Aedes arborealis - 1 1 2 - 1 - - - - - - 5 6
argyrothorax - 1 - - 1 - - - - - - - 2 6
fulvithorax 1 1 3 2 - 1 4 3 2 1 1 1 20 19
fluvus 3 4 1 - 3 1 - - - 6 1 - 19 14
scapularis 11 26 25 11 6 2 5 17 111 135 27 253 629 800
septemstriatus 13 - 8 1 2 4 8 6 2 - - 2 46 43
serratus 106 14 5 1 - 10 24 38 57 176 25 13 469 502
Culex spp. 52 46 36 10 33 60 38 16 79 2 15 12 329 294
B 1 6 1 7 3 1 - - - - - - - 18 12
B 7 10 10 - 1 8 5 - 3 5 - 4 2 48 67
B 8 - 2 - - - 2 - - - - - 5 9 11
B 9 176 87 63 32 68 61 20 8 45 11 39 18 628 494
B 17 6 7 40 23 11 16 3 - 9 - - 1 116 106
B 19 76 61 18 18 56 75 26 46 92 122 148 74 812 847
(Culex) coronator? 18 7 2 2 1 - - - 5 - 10 5 50 67
pipiens - - - - - - - - - - 1 - 1 0
declarator 3 - - - - - - - - 1 - - 4 12
(Melanoconion) spp. 10 4 1 1 - - - - - - - - 16 29
caudelli? 3 - - - 1 2 - - - 1 2 - 9 0
spissipes 86 83 25 9 6 15 - 1 2 - 1 1 229 121
taeniopus 68 48 27 44 84 77 7 2 21 99 67 15 559 527
Haemagogus spp. - - - 1 - - 1 - - 1 - - 3 2
Mansonia spp. 33 99 81 90 83 174 221 142 47 - 18 2 990 1138
albicosta 2 - 1 2 1 2 - 2 - - - - 10 8
arribalzagai 95 70 77 38 35 44 35 53 59 24 23 2 555 529
titillans - 1 - 3 2 3 4 11 11 - 4 - 42 44
venezuelensis 160 287 248 521 491 162 372 278 201 3 35 4 3062 3414
Psorophora albipes 52 14 1 - - - - - - - - 1 68 57
cingulata - - - 1 - - - - 1 1 2 - 5 5
ferox 9 2 - - 1 - 12 15 6 - - - 45 39
lutzi 2 - - - - - 1 - - - - - 3 2

Table 23 (continued)
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Total Varzea
total
SABETHINI
Limatus flavisetosus 5 1 1 - - 2 - - - - - - 9 6
paraensis - - - - - 1 - - - - - - 1 2
Phoniomyia spp. - - - - - 1 - - - - - - 1 4
Trichoprosopon digitatum 1 6 23 19 31 15 11 5 2 3 1 5 122 120
edwardsianus - - - - - - - - - - 1 - 1 1
magnus 1 - - - - 1 - - - - - - 2 0
Wyeomyia spp. 1 - 2 - - 4 1 - - - - - 8 8
(Dendromyia) aporonoma - - - - - 5 1 - - - - - 6 5
ANOPHELINI
Anopheles (A.) intermedius 4 2 2 - 1 - 1 - 1 1 - - 12 12
(A.) mediopunctatus 1 - 1 2 1 1 1 22 9 - - - 38 36
(N.) oswaldoi - - - 1 - - - 1 6 - 3 1 12 15
(N.) triannulatus - - - - - 1 - 1 7 2 1 - 12 9
(S.) nimbus 1 - 2 7 15 40 36 3 - - - - 104 58
T o t a l 1015 885 701 845 942 1088 832 676 710 589 429 417 9139 9494
Capture time (hours) 15:00 10:25 8:20 12:00 8:00 10:00 8:00 8:00 10:00 8:00 8:00 9:00 114:45 114:45
Rate per hour 68 85 84 70 118 109 104 85 71 74 57 46 80
Rate per hour (varzea) 55 78 72 86 115 92 96 101 90 65 62 64 83
Rate per hour (Utinga, day) - 111 160 163 97 125 72 71 72 110 87 59 99

Table 24
Daytime catches on human bait at ground level Serra do Navio, Amapá, 1965
Jan Feb Mar Apr May Jun Jul Aug Sep Oct§ Nov Dec Total
CULICINI
Aedes arborealis 1 21 9 25 22 119 86 26 1 310
argyrothorax 1 2 3
fulvithorax 1 7 9 25 68 69 34 7 2 1 224
fluvus 34 16 14 7 248 70 15 1 405
leucocelaenus 3 1 2 12 5 1 24
scapularis 88 37 42 114 36 8 325
septemstriatus 1 1
serratus 13 388 143 157 344 144 86 57 7 5 1344
Culex spp. 1 3 3 2 1 10
B 7 1 1
B 9 1 1
B 17 3 3
B 18 1 1
B 19 1 1
(Culex) coronator? 12 4 1 2 1 10
(C.) declarator 2 1 3
(Melanoconion) taeniopus 1 1
(Microculex) stonei 3 3
Haemagogus spp. 4 4
Mansonia spp. 6 6
arribalzagai 6 14 6 25 43 29 14 16 2 4 2 161
venezuelensis 12 12
Psorophora albipes 1423 107 324 169 2177 29 3 4232
ferox 5 276 50 45 113 71 42 6 608
SABETHINI
Limatus spp. 10 11 6 1 1 29
flavisetosus 31 8 13 22 11 30 6 1 5 127
paraensis 2 2 6 4 13 8 1 2 38
pseudomethisticus 4 1 5
Phoniomyia spp. 13 15 9 5 26 4 11 13 2 4 1 103
Sabethes (S.) spp. 1 4 3 8
(S.) albiprivus 1 1
(S.) belisariori 1 2 3
(S.) cyaneus 1 4 2 1 1 9

Table 24 (continued)
Jan Feb Mar Apr May Jun Jul Aug Sep Oct§ Nov Dec Total
Trichoprosopon digitatum 4 1 23 9 8 55 7 13 120
magnus 5 3 6 32 5 14 24 1 2 3 95
Wyeomyia spp. 250 360 149 161 477 229 283 177 13 71 6 2176
(Dendromyia) aporonoma 3 10 7 5 15 8 6 10 6 1 71
melanocephala 3 3
ANOPHELINI
Anopheles (A.) eiseni 1 1 2
(A.) darlingi 1 1
(A.) intermedius 1 1 1 3
(A.) mediopunctatus 2 2 2 5 2 2 15
(N.) oswaldoi 1 2 2 5
(S.) nimbus 5 3 4 1 1 14
T o t a l 342 2677 563 873 1515 3186 746 435 38 128 18 10521
Capture time (hours) 11:15 27:00 6:00 10:30 31:00 47:00 50:50 55:50 5:30 25:00 7:00 276:55
Rate per hour 31 99 94 83 49 68 15 8 7 50 3 38
Rainfall (mm.) 222 226 354 131 396 119 110 180 84 57 51
§ October catch spoiled.

-95-
Table 5
Daytime catches on human bait at Porto Platon, Amapá, April 1965
Km. 162
Ground Tree
Campo Verde
Ground Tree
SubTotal
Ground Tree Total
CULICINI
Aedes arborealis 2 2 2
fluvus 2 2 2
leucocelaenus 1 5 5 5 6 11
scapularis 2 27 29 29
serratus 2 21 23 23
Culex spp. 1 1 1
(Culex) declarator 3 3 3
Haemagogus spp. 32 15 169 15 201 216
Mansonia arribalzagai 2 2 2
Phoniomyia spp. 1 3 4 4
Psorophora 104 329 8 433 8 441
ferox 10 41 1 51 1 52
lutzi 1 1 1
SABETHINI
Limatus spp. 5 5 5
flavisetosus 27 41 68 68
paraensis 6 6 6
pseudomethisticus 3 1 4 4
Sabethes (Sabethes) spp. 2 1 1 2 3
(S.) belisarioi 1 1 6 1 7 8
(S.) cyaneus 1 2 15 2 16 18
(Sabethoides) chloropterus 5 1 13 1 18 19
(S.) glaucodaemon 1 1 1
Trichoprosopon digitatum 1 1 1 1 2
magnus 1 1 1
Wyeomyia spp. 17 28 4 45 4 49
(D.) aporonoma 27 27 27
ANOPHELINI
Anopheles (A.) mediopunctatus 2 2 2
T o t a l 167 42 567 224 734 266 1000
Capture time (hours) 3:30 3:30 26:00 19:30 29:30 23:00 52:30
Rate per hour 48 12 22 11

1
Table
Mbsquitoes taken by Mk.I b1ower trap on various baits at 6 mohigh, utinga forest,
January, 1965
CULIGINI
Aedes fu1 vi thorax
i:ulvus
hortator
1eucoce1aenus
septemstriatus
serratus
CUlex spp.
B 7
B 9
B 17
B 18
B 19
(Culex) dec1arator
(Me1anoconion) spp.
(M.) spissipes
(Mo) taeniopus
Haemagogus spp.
}Msonia spP.
arriba1zagai
titi11ans
venezue1cnsis
Ps.orophora a1bipes
ferox
1utzi
Uranotaenia hystera
SABETHllfr
Lirrl:L tus fiavisetosus
Pnoniomyia spp.
Sabethes (Sabethes) spp.
(S o) arll8.zonicus
(S.) be1isarioi
(So) cyaneus
(S.) quasicyaneus
(Sabethoides) chloropterus
(S.) glaucodaem on
Trichopro.sopon digitatum
magnus
i'lyeomyia spp.
(D.) aporonoma
J.t>use
26
1
1
1
9
1
10
7
4
10
4
5 2
2
2
2 2
4 5
32
1
1
6
411
6
3
8
3
2
1
1
3
2
4
5
2
5
12
6
2
3
613
16
1
9
38
15
7
15 1
1
3
1
22
188
33
1
1
1
Total 60 31 6 25 25 996 85
1
2
1
1
10
5
1
14 2
4
32
13 3
20§§
II Q da~rs 3 3 2 2 2
Rate per day 20 10 3 6 13 50 43
§ hand capture for com~rison
§§ average of only 7 hrso catching per day, by daylight
4
1
1
1

Table 27
Mosquitoes and Phlebotomus taken in blower traps on different baits at ground level, IPEAN forest, 1965
Cebus
(Dec)
Saimiri
(Nov)
Guineapig
(Sep)
Rabbit
(Aug)
Ameiva
(Jun)
Honey
(Apr)
Proechimys
(Apr)
Mouse
(Apr)
Bird §
(Mar)
Chicken
(Feb/Oct)
Tamarin
(Feb/Dec)
CULICINI
Aedes arborealis 2 4
argyrothorax 1
fluvus 3 1
hastatus 1
hortator 1
scapularis 1 87 1 11
septemstriatus 1 2
serratus 1 20 1 3 1 4
Culex spp. 8 35 6 221 321 21 6 1 30 8
B 7 3 28 3 126 92 1 4 12
B 8 1 1 2 1
B 9 18 109 8 253 262 1 8 26 1 60 48
B 17 38 2 496 453 11 3 1 3
B 19 3 28 9 7 19 22 7
(Carrollia) spp. 1
(Culex) spp. 10 1
(C.) coronator? 5 1 1 2
(C.) declarator 12 1 3
(Melanoconion) spp. 10 5 7
(M.) caudelli? 2 29 66 161 37 3 6 5
(M.) spissipes 16 66 1 9 7 1 1 1 10
(M.) taeniopus 54 490 15 176 127 4 4 24 2 146 131
Mansonia spp.
arribalzagai 3 3
titillans 1
venezuelensis 1 5 2
Psorophora ferox 2 2
lutzi 6 9
Uranotaenia calosomata 1

Table 27
Cebus
(Dec)
Saimiri
(Nov)
Guineapig
(Sep)
Rabbit
(Aug)
Ameiva
(Jun)
Honey
(Apr)
Proechimys
(Apr)
Mouse
(Apr)
Bird §
(Mar)
Chicken
(Feb/Oct)
Tamarin
(Feb/Dec)
SABETHINI
Limatus durhami 1
flavisetosus 2 2 1 1 2 2
paraensis 1 6 1 33 1 6
Phoniomyia spp. 1
Trichoprosopon digitatum 1 1 2 1
magnus 1 1
Wyeomyia spp. 3 3 4 3 1 2 4
(Dendromyia) aporonoma 2 3 2
PSYCHODIDAE
Phlebotomus spp. 1 5 2 2
Total 128 998 36 1370 1484 17 82 89 5 287 268
N° of days 9 16 2 2 2 2 3 8 6 15 9
Rate per hour 14 62 18 685 742 9 27 11 1 19 30
N° of species 20 26 7 15 18 4 12 12 4 16 22
§ Pipra sp. and a hummingbird (exposed one day each, died during exposure).

Table 28
Mosquitoes & Phlebotomus taken in MK.II blower traps on different baits at ground level, with times of capture,
IPEAN forest, 1965 §
Times of capture
October November December December
July through December
B a i t
Chicken Saimiri Tamarin Cebus 0830 1330 1930 0130
to to to to
(fed) total (fed) total (fed) total (fed) total 1330 1930 0130 0730
CULICINI
Aedes argyrothorax 1 1
fluvus 2 2 2 1
hastatus 1 1
hortator 1 1
oligopistus 1
scapularis (5) 84 1 1 11 13 20 15 21
septemstriatus 2 2
serratus 20 1 1 (1) 4 7 18 12 8
Culex spp. (1) 27 30 3 8 9 39 17 35
B 7 (1) 14 4 2 12 5 9 31 20
B 8 1 (1) 1 3 1 1
B 9 (6) 77 (3) 60 (3) 8 (15) 48 39 104 135 144
B 17 1 1 3 4 1 7 3
B 19 (2) 26 22 3 (2) 7 16 49 21 32
(Culex) spp. (1) 6 1 1 3 2 1
(C.) coronator? 5 2 1 1 3 2
(C.) declarator (2) 12 3 1 3 7 3
(Melanoconion) spp. 10 5 (1) 7 2 7 12 3
(M.) caudelli? 14 6 5 5 21 10 8
(M.) spissipes (2) 22 1 10 8 24 6
(M.) taeniopus (10) 445 (3) 146 13 (20) 131 18 244 372 226
Mansonia spp. 2
arribalzagai 1
titillans (1) 1 1
venezuelensis 1 2 9 11
Psorophora ferox (1) 2 1 1
lutzi 1 1
Uranotaenia calosomata 1

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Table 29
Mosquitoes by kind and month captured under hood with sentinel mice, Utinga forest, 1965
31 28 31 30 31 30 31 31 31 334 365
Jan Feb Mar Apr May Jun Jul Aug Oct Totals
N° of captures (days)
CULICINI a) b) c) d)
Aedes scapularis - - - - - - - - - - 3
- - - (1) - - - - (2) (3)
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- - - - (1) - - - - (1) 2
serratus - 1 - - - 2 1 - - 4
- - - (1) (1) (2) - - - (4) 8
Culex spp. 53 54 45 18 19 19 5 - - 160
656
- (96) (95) (90) (92) (63) (7) - - (443)
B 1 9 1 12 10 - - - - - 23
- (13) (45) (31) - - - - - (89) 121
B 7 56 14 19 7 - 2 - 1 - 43
- (18) (19) (3) (3) - - - - (43) 142
B 8 3 - - 2 - - - - - 2
- (1) (3) (3) - (1) - - - (8) 13
B 9 108 43 41 25 16 3 2 - - 130
(47) (25) (4) (7) (5) - - - (88) 326
B 17 5 6 4 4 3 - 1 - - 18
- (2) (4) - (2) (1) - - - (9) 32
B 19 15 8 7 3 - 2 - - - 20
- - (3) (1) - - - - - (4) 39
(Culex)coronator? 5 - - - - - - - - -
- (4) - - - - - - - (4) 9
(Melanoconion) spp. - 5 14 8 - - - - - 27
- (2) (8) (10) - - - - - (20) 47
(M.) caudelli? 2 - - - 7 1 - - - 8
- - - - (1) - - - - (1) 11
(M.) spissipes 13 5 2 1 - - - - - 8
- (2) (2) - (1) - - - - (5) 26
(M.) taeniopus 12 3 3 1 1 1 - - - 9
- (7) (2) - - - - - - (9) 30
(Microculex) stonei - - - - 1 - - - - 1
- - - - - - - - - - 1

Table 29 (continued)
31 28 31 30 31 30 31 31 31 334 365
N° of captures (days)
Jan Feb Mar Apr May Jun Jul Aug Oct Totals
a) b) c) d)
Mansonia spp. - 1 - - - - - - - 1
- - - (1) (1) (3) - - (5) 6
arribalzagai - - - 1 - - - - - 1
- - (6) (1) - - - (1) (8) 9
venezuelensis - - 1 - - 1 - - - 2
- (1) (4) - (5) - - - (10) 12
Psorophora lutzi - - - - - - - - -
- - (1) - - - - - (1) 1
SABETHINI
Limatus paraensis 1 - - - - - - - - 1
- - - - - - - - - 2
Trichoprosopon digitatum - - - - - - - - - -
- - (1) - - - - - (1) 1
Wyeomyia spp. 1 - - - - - - - -
- - (2) (2) (3) - - - (7) 8
(D.) aporonoma - - - - - - - - - -
- - - (1) - - - - (1) 1
Subtotal: fed - 141 149 81 47 31 9 1 - 459
unfed (192) (207) (158) (113) (81) (10) - (3) (764)
Totals 283 333 356 239 160 112 19 1 3 - 1560
a) fed mosquitoes not separated from unfed in January
b) during rest of year, unfed totals given in parentheses below fed totals
c) no mosquitoes found in Sep., Nov., Dec.
d) Subtotals omit January figures

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Table 30 (continued)
31 28 31 30 31 30 31 31 30 31 31 334 365 365
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Dec Totals Utinga
N° of captures (days)
3 -
132 47
42 11
30 26
87 30
11 6
10 9
1 -
18 12
(C.) declarator 1 - - - - - - - - - -
- (1) - - (1) - - - - - (2)
Culex (Melanoconion) spp. 5 15 31 13 - - - - - - - 59
(8) (31) (29) - - - - - - - (68)
(M.) caudelli? 8 - - - 12 12 3 - - - - 27
- - - (5) (2) - - - - - (7)
(M.) spissipes 18 1 1 - 4 2 - - - - - 8
- (2) - (1) (1)- - - - - - (4)
(M.) taeniopus 18 6 10 3 6 8 7 2 - 2 - 44
(4) (4) (2) (6) (5) (2) (1) - - (1) (25)
Mansonia spp. - - - - - - - - - - -
- - - - (8) (1) (2) - - - (11)
arribalzagai 1 - - - - 1 - - - - - 1
- (1) (2) - (1) (1) (2) - - (1) (8)
titillans - - - - - - - - - - - -
- - - - - - - (1) - - (1)
venezuelensis - - - - - - - 1 - - - 1
- - (6) - (1) (3) (7) - - - (17)
Psorophora ferox - - - - - - - - - - -
- - (1) - (1) (1) (1) - - - (4)
lutzi - - - - - - - - - - - -
- - (1) - - - - - - - (1)
SABETHINI
Limatus durhami - - - - - - - - - - -
(1) - - - - - - - - - (1)
flavisetosus - - - - - - - - - - - -
- - (1) (2) (1) - - - - - (4)
paraensis 7 - - - - - - - - - - -
(3) (2) (1) (1) - - - - - - (7)
Trichoprosopon digitatum - - - - - - - - - - -
- - - - - - - - - - -
- - - (2) - - - - - - (2)
Wyeomyia spp. - - - - - - - - - - - -
(1) - (4) (7) (10) (2) - - - - (24)
(Dendromyia) aporonoma 2 - 1 - - - - - - - - 1
- (1) - (1) - - (2) - - - (4)
4 -
1 1
1 -
4 -
14 2
2 1
24 8
7 1

Table 30 (continued)
31 28 31 30 31 30 31 31 30 31 31 334 365 365
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Dec Totals Utinga
N° of captures (days)
1 -
ANOPHELINI
Anopheles (S.) nimbus - - - - - - - - - - - -
- - - (1) - - - - - - (1)
Subtotals - 214 226 107 113 110 73 19 - 7 - 869
(357) (418) (257) (348) (286) (143) (44) (3) (14) (11) (1881)
3589 1506
Total 1109 571 644 364 461 396 216 63 3 21 11 -
1506
Utinga total 283 333 356 239 160 112 19 1 0 3 0 -
as for Table 29
a)
b)
c)
d)

Table 27
Mosquitoes taken in blower traps on mouse baits at ground level, IPEAN forest, 1965
Total
Feb Mar Apr Jun Jul Aug Sep Oct Nov Dec
Station I I I II I II II II II II II
CULICINI
Aedes arborealis 7 2 5 1 15
fluvithorax 6 2 1 9
fluvus 6 2 1 2 2 1 14
hastatus 1 1
oligopistus 1 1
scapularis 11 5 17 1 1 4 1 6 1 16 63
septemstriatus 1 1 2
serratus 11 1 1 9 13 12 3 11 1 62
taeniorhynchus 2 1 3
Culex spp. § 1423 582 802 221 247 206 118 23 78 38 54 3792
B 7 307 207 241 126 33 117 288 8 73 23 63 1486
B 8 31 49 110 28 7 2 1 1 229
B 9 § 1095 815 1336 253 255 178 513 73 188 150 207 5063
B 17 § 789 570 1420 496 138 171 55 3 13 3 3658
B 19 § 112 54 53 9 70 16 89 14 39 46 15 517
(Culex) spp. 57 57
coronator? 7 9 6 1 1 4 4 3 2 1 38
declarator 2 1 1 4
(Melanoconion) spp. 7 2 26 11 18 64
caudelli? 221 140 241 66 80 32 44 3 16 3 3 849
spissipes 601 129 131 9 18 9 18 9 10 934
taeniopus § 1065 399 852 176 216 158 354 71 637 565 250 4743
Mansonia spp. 6 4 2 6 2 20
arribalzagai 1 14 8 1 3 27
titillans 2 1 3
venezuelensis 36 3 13 5 10 11 26 1 1 1061
Psorophora albipes 1 1
ferox 4 1 5
lutzi 15 1 16

Table 31 (continued)
Total
Feb Mar Apr Jun Jul Aug Sep Oct Nov Dec
Station I I I II I II II II II II II
SABETHINI
Limatus durhami 8 6 14
flavisetosus 4 14 7 1 1 27
paraensis 17 30 23 1 4 1 1 77
Sabethes (S.) quasicyaneus 1 1
Trichoprosopon digitatum 10 3 16 1 30
magnus 1 1
Wyeomyia spp. 23 20 39 4 8 1 1 1 97
(Dendromyia) aporonoma 5 3 2 2 2 14
ANOPHELINI
Anopheles (A.) intermedius 3 5 8
(A.) mediopunctatus 1 1
(A.) oswaldoi 1 1
(S.) nimbus 1 1 1 3
Identified total 5887 3066 5343 1370 1129 931 1541 204 1101 844 640 22,056
Catch total §§ 8388 7563 11664 6643 4632 2791 1541 204 1101 844 640 46,011
N° of days 15 19 17 10 18 13 17 14 17 15 19
Rate per day 559 398 686 664 257 215 91 15 65 56 34
§ some of these spp. were used for breeding and transmission studies.
§§ some mosquitoes were counted but not identified for lack of time.

Table 32
Number of mosquitoes taken by MK.I blower trap on mouse bait according to fan cycle
(09 secs. Every 15 or 30 mins.)
Jul Aug Sep Oct Nov Dec Total
(mins.) 15 15 30 15 30 15 30 15 30 15 30 15 30
(days) 3 8 9 4 10 9 8 6 9 11 8 41 44
Fan interval
Captures
CULICINI
Aedes fluvus 1 1 1 1 2
scapularis 1 2 2 1 3 3 1 13 3 20 9
serratus 13 10 2 1 2 7 4 1 32 8
Culex spp. 206 93 25 12 11 50 28 19 19 30 24 410 107
B 7 117 222 66 4 4 44 29 13 10 35 28 435 137
B 8 7 2 1 1 11
B 9 178 356 157 30 43 102 86 63 87 124 83 853 456
B 17 171 29 26 3 8 5 3 211 34
B 19 16 54 35 2 12 22 17 22 24 12 3 128 91
(Culex) coronator? 4 3 1 1 2 2 1 11 3
declarator 1 10 16 8 3 16 2 34 21
(Melanoconion) spp. 32 24 20 3 10 6 2 1 1 2 69 32
caudelli? 9 13 5 3 6 7 3 32 14
spissipes 158 214 140 32 39 308 329 252 313 157 93 1121 914
taeniopus 2 4 2 2 6 4
Mansonia spp. 3
3
arribalzagai 1 1
titillans 11 21 5 1 1 33 6
venezuelensis
SABETHINI
Limatus flavisetosus 1 1
paraensis 1 1 2
Wyeomyia spp. 1 1 1 3
(Dendromyia) aporonoma 1 1 1 1
ANOPHELINI
Anopheles (A.) mediopunctatus 1 1
(N.) oswaldoi 1 1
(S.) nimbus 1 1
Subtotals 931 1052 489 83 121 569 532 387 457 399 241 3421 1840
Total 931 1541 204 1101 844 640 5261

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Table
36
Fema.le mos qui toes I an1 PhlebotollnlS caught wi th hand suction trap on fo1iage
in IPEAN forest, September through December 1965
OULIOnlI
Aedes horta ror
oligopistua
scapularis
serratus
C\11ex 8pp.
B 7
B 8
B 9
B 19
B 21
(Carro1lia) sp.
(Culex) 8pp.
(O.) coronator?
(O.) declarator
(I~lanoconion) spp.
(M.) caudelli?
(M.) spissipes
(Mo) taeniopus
(Microculex) B 1
B 2
Imsonia spp.
arribalzagai
tiUllans
venezuelensis
Orthopodomyia fas c i pes
Psorophora ferox
Uranotaenia spp.
cooki
hystera
leucoptera
SABE'rHIN I
Limatus flavisetosus
paraensis
Trichoprosopon digi ta tum
Wyeonvia spp.
PSYCHOD mAE
L;aPOI3.r&
M/F-
1 O
116
2l
4
1
11
.5
O
1
8
63
64
14
33 O
4
O
O
O
O
O
O
O
O
O
1
O
1
o
o1
O
2
O
30
9
38 6
1
13 2
O
O
3
24
10 4
111
181O
6
1
2
29 O
O
1
1 O
O
o o
o
1
9 O
2
13
12
5
o
li
3
1 O
16
36
964
16 O
13 1
2
O
O
O
O
1
O
O
1
1
O
o1
3
1
3
1
25 7
55
12 1
13 4o2
12
22
38 1
22 1
52 2
1
O
O
O
O
O
1
O
1
O
O
311
1
10 O
3
29
33 91
22
8
1
1
24
99
160
18
49 o
17 12O
O
O
O
1
O
O
2
1
1
o
1
4
1
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s~ I
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93 i
18
2~ I
6
O
2
15
~
48 S
33 2
70
3
1 61
2~ I
O
1
1 2O
O
3
1
1 2
1.,
1.
,8
45
126
27 3
48
14 1
3
39
14,
208
23
82 2
H11ebotomus spp. o 16 o 24 o 40 40
Subtotals
Totals
.,
Capture tjJne (hours)
87 4361
2
29 11
1
4
1
1
3 2
.5
3
.1.034
,1034
24 120
Ra te per hour 113
§ 2 haure ot f_l. capÜ1reS accidentaliy diStroyec1.
§

115 -
Table 37
*1e l11lebotOllUB captured b)" suctim apparatus on tree-~ 1n Utinga forest, 1965
-
P. antunesi
P. aragaoi
P. baityi
P.carvalhoi
P. coutinhoi
P. damascEmoi
P. dm1drol:1h1'llus
Pó .f'urc atua
P. infraap1nosus
P. intermediU8
P. longi8p1nU8
P. lutz1anU8
P. p1losua
P. pinotti
P. rorotaeDSis
Pé scaff1
P. sham1cmi
P.súis
P. triacantJlU8
P. trisp:1nOSU8
P. tubercula tua
P. ubiqui talis
P. villelaiP.
wiJ1 jll~i
P. sp. nov. ?
}.By Jw1 Jul Aug Sep ~t Nov Dec To~
126
-
--
46
11
-
-
- -
-
54
-
4
10
-
194 2
S
-
,)
-
- 1
- .90
22
29
-
- -
140
1
3
17
-
60 2
-
-
-
48
-
.
8
-
S7
12
--6----
114
-
.3
20
-
-
43 4
-
-
4
24 -
.96
-
79
18
-32
- -
-
67 2
1 8
- 41
- -25
93
-
31
-188
20
4
6
-
-
-
-
2)0
2~ --1
-
183
70
8
-1
-
-
-
-
658
4
14
22
-
-
27
4 -
-1
103
-
-
23
- 70
U--
1
-
253
14 1
4
1
53
38
--
32 11-
1027
~1
164
1
766
232
4l9
10
32111
-221
-
4
10
-
10
4
-
4
.3
2
24
--
3 -
-
-
4
320
-5
20
2
2
8
-
112
1804
10
36
131
2
2
369
17
62
4.3
Total 4SS 546 331 604 ~ 49, 759 4722
Capture t1m (hours) , 130 11130 10..308130 91)0 8130 8130 9:00 n 1)0
Ra te per hov 83 47 30 30
63 58

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Jan.22
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7
20
22
23
23
Jul,10
12
12
13
13
13
14
19
20
20
20
20
-
-117 -
Table 39
Blood parasites found 1n wild vertebrates,
utinga
" n
IPEAN
M
Utinga
"
" U
AN 147.38
AN 14757
AN 14764
AN 15'025'
AN 15'026
AN 15038
AN 15'046
AN 14302
AN 14418
1965 §
(M4.) trypanos ame
D s (~) IIdcrof'ilaria
microfilaria
5 IPEAN AN 15096 ~OBa murina (MA.
..
li
" li
"
"
"
li
li
li
U
-
199371
199380
199435
199445
199449
199458
199.518
199696
199746
199747
199758
199760
33161
_Tamarin ~rin (PR) tryp. & microfil.
" " trypa.nosome
DidelphiB marsup!al!s (MA' trypanosome
Tamarin tamarin (m) tryp.& microf.
Calluromws' Bp. (MA) microfilaria
?o'i1rmosa sp. (MA" tryp. & microf.
.
microfilaria
Kentropyx striatu~ (RE) microfil.& Plasmod.
,Turdus fUJTd.g~tu~ (AV) microfilaria
~Plica umbra (RE) rn1crofila.ria
cTurdus albicollis (AV) microfilaria
Glyphorhynchus spi!:!!!:!:!~ (AV) haemogregarine
Arremon taciturnus (AV) Plasmodium
Ramphocelus carbo (AV) Plasmodinm -
Arremon taci turnus {A V )m1crofil.& Plasmod.
'l'urdus albicollis (A V) trypa.nosome
~Formicarius analis (A V) rn1crofilaria
Ramphocelus carbo (AV) Plasmodium
li li li
(bat) Bartonella sp.
§ the bird, bat & lizard bloods were examined by the Wellcome Parasitology Unit
a t the Ins ti tu to Evandro Chagas.
(AV) .Bird, (MA) .marsupial, (PR) .primate, (RE) .lizard.

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Table lu
V1Z'a8.. 18o1&ted; b't& 8_~-.-!!8l .s.e. 1n adjacmt baI'88
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71&83
a! 7$27
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Fig. 5.
126
The new meteorological station at the IPEAN forest.

Fig.
6. The new entomology laboratory in the IPEAN forest.

Fig. cages, intervals.
128 -
7. The Mk. II blower trap showing the 4 collecting
and the clock which actuates the fan below at regular

-129 -
Fig. 8. The Mko 11 blower trap showing the second clock
with descending plunger which makes contact every 6 hours to run
a winch which moves the fan assembly to another cage. Also shown
are the relays ~nd the plastic-covered ant-proofed battery.

APPENDIX I
TRE SEROLOGICAL RESPONSE OF ANlMALS TO VIRUS INFECTION
IN UTINGA FOREST
A mammal recapture program has been carried out in Utinga forest,
Be1em, Brazi1 since June, 1962 (Causey, C.E. 1963). As tropical rodents
and marsupials were trapped, marked, and released, blood specimens were
taken for virus isolation attempts in baby mice. At 2 to 3 week intervals,
returning animals were studied serologically,
Hemagglutination-inhibition (RI) tests were performed on acetone
extracted sera by techniques previous1y described (Shope, Robert E. 1963).
The viruses used are 1isted in Tab1e 1.
Neutralization (N). testing was performed in baby mice using either
mouse serum or brain as virus source, incubating the serum-virus mixture
for 1 hour at 37°C, and inoculating either intracerebrally (IC) or intra-
peritoneally (IP).
with approximate1y 100 LDSO of viru9.
Animal sera were tested in a final dilution of 1:4
Results were interpreted as
negative if a11 mice died, and as positive if 4 or more out of 6 survived
Eighty-five forest-acquired virus infections were studied over a
3 year period with serological determinations in serial bleedings. lu
every animal, virus was isolated from the blood.
Figure I shows the RI responses of animaIs infected with Mucambo
virus of group A and Bussuquara virus of group B. In 104 b1eedings prior
to, or at the time of viremia, no inhibition was detected. ln one animal
the initia1 b1eeding 31'2 months before infection, had a titer of 1:10
and the possibility of this representing maternal antibody is raised.
bleedings following cessation of viremia contained HI antibody which per-
sisted for the duration of observation (425 days in the longest observation).
Initial titers were higher than later ones. Testing with Mucambo virus
in 4 animals confirmed the presence of N antibody following infection!
All

ses. uated.
Appendix I
132 -
Figure 2 shows the RI response of animais infected with group C
and group Guama viruses. In 134 b1eedings prior to, or at the time of
viremia, no inhibition was detected. One animal had a 1:10 reaction at
the time of Guama viremia. RI antibody was not detected in 6 animaIs
following viremia. ln contrast to the response after infection with
viruses of groups A and B, HI antibody in most of the animals became
either non-detectable, or was apparently lost and restimulated, or
This fluctuation might be explained by variation in sensi-
tivity of antigen from test to test, however, an alternative explanation
is that RI antibody to groups C and Guama is short-lived and is restimu-
lated by recurrent natural exposure of the animal to the same or related
Table 2 shows results of testing of a Proechimys after Guama
infection in which a response to both Guama and Moju antigen was detected
then lost. Guama antibody reappeared with Capim antibody, then again
diminished to rise later accompanied by a generalized Guama group reaction.
Sero10gy for Bussuquara virus, a180 iso1ated from this garoe animal, is
included to illustrate the consistency of response in group B
N testing with 4 anima1s infected with Caraparu, 10 with Guama,
2 with Catu, and 1 with Capim virus showed uniform development of N anti-
body following viremia
The HI response of animaIs infected with phIebotomus fever group
viruses is shown in Figure 3. As with groups C and Guama, the RI antibody
leveI fluctuated and sometimes became non-detectable
Table 3 shows N test resulte with Acará and Pacui viruses. All
animals developed significant N antibody following viremia.

al.
I
-133 -
These results offer a firm basis for interpretation of RI antibody
in surveys of wild animals iu Utiuga forest: firstly, non-specific re-
actions are not anticipated since bleedings prior to infection were
generally negat1ve; secondly, RI ant1body to Mucambo and Bussuquara v1rus
following viremia may be expected to persist for the life of the animal;
and thirdly, RI reactions following infection with viruses of groups C,
Guama and phlebotomus fever are likely to represent recent primary in-
fect1on or anamnest1c responses to exposure to a virus in the same sero-
logical group and this antibody may not persist for the life of the
2.
References
Ca1ista E., The role of sma11 mamma1s in maintenance of
arboviruses in the Brazi1ian Amazon forests. (1963) Anais de
Microbio1. !!, Parte A, 119-121.
Robert E., The use of a micro hemagg1utination-inhibition
test to fo11ow antibody response after arthropod-borne virus
infection in a community of forest anima1s. (1963) Anais de
Microbio1. ~, Parte A, 167-171.

Appendi:x: I
Group
A
Virus
Mucambo
B Bussuquara
c
Guamá
Ungrouped
Phlebotomus
fever
Caraparu
Nepuyo
Murutucu
GuamÁ
Catu
(Moju)*
(Capim)
(Acara)
(Pacui)
Itaporanga
(Bujaru)
Icoaraci
-134 -
Table 1
Viruses Used in Serological Testing
Strain
Be An 10967
Be An 4116
Be An 3994
Be An 10709
Be An 974
Be An 277
Be H 151
Be Ar 12590
Be An 8582
Be An 27639
Be An 27326
Be An 64582
Be An 47693
Be An 24262
HA
Preparation
brain
brain
serum
serum
serum
serum
serum
serum
brain
-
brain
brain
brain
N Test
Virus Source Route
serum
serum
serum
serum
serum
serum
brain
* Viruses in parentheses are unpublished and mention here does not constitute
formal description.
1.p.
t.p.
t.c.
i.c
i.c

5
Appendix I
Bleeding Date
Table 2
HI testing of Proech1mvs RO 5889 with group Guama viruses
RI Results
Group B Group Guamá
Vires
-
Isolated Guamá Moju Cat6 Capim
22 IV '64 o o o o o
6 V
21 V o
26 V Bussuquara
12 VI
2 VII
22 VII
7 VIII
25 VIII
18 IX
Guamá o o o
o o
40 40 o o
40 o o o o
)80 80 o o 10
40 40 o o o
40 40 o o o
40 10 o o o
40 80 20 10 o
80 40
20 40 o
80 40 20 20 10

Appendix I
-136 -
Table 3
Results of N Te8ting with Ungrouped Virule8
Genus Viru818o1ated Log Neutralization Index
Pre Post Days after viremia
Nectomvs Acará 2.2 10
Pacui 2.2 22
>2.1 49
Pacui 2.9 15
>2.9 97
Pacui 2.2 19
Pacui 3.1 15

[
Appendix I
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H O
p, ro
Q) fJ) H
H ~ ro
O .~ g.
...'d ~
Q) fJ)
t> Q)
ro M
~ ,Q
~

Appendix I
U)
w
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a
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lL a
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u
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Pt & ro ~"'
ro :;j M
~ d M i
ro Q) ro
ozOoo
-to' N
o"' M
-to' 1 ~~ "' ~
:>. ~
'O o 'n ~ -..t-
P-to'Q) o o ro ~
'n o M UJ
~ :.%1 '" ~ ~ "
ro M °nI I'n Q >
l
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~ oo~o ~ "'~t
"n Q) -to' UJ ~
-to' o
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~ ~ Q) ~
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-to' 1 > 'O Q) ';:j 1 ~ ~> 8
O o, -..t- ,;-I
ro N ;:j Pt
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Appendix I
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a:
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'--" ,~ H Q) (/J
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(/J H H
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(/J ro
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H O Ç1
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H bD O
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ti Q)
ai M H
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(/J

[I Due ~~us t.he o:f. ~ternocm. Prefeito. 1'be order D1ey Frei 1\1rÚS 1. ror 'l'he nines Arrani;e..~'.ts
p'!!~-p~t1on
!ao There
~PPENDIX
REroRT W THE IAmFA RmICtl EPIDEMIC
Sept8li>er .Hovember 1965
to u.anB}X>rtatim difticult1es, tlle visiting group wa:s delayed 111
for 'talo dan. Visita vere ~"d to FSFBP headquartere J to the Governar,
Se=-etar.v aí Health and to tl'.e Instituto Nacional de Pesquisas. .An aaJp111b1an
t-he FAB arriVM on the 16th oí October and !abres. was reached tl1at s~
Oontacts were establ1shed with the autllorities, consüting 0.1" ao
a Pr~tor arn a Delegado.
vere at the Pier to receive t.he cODln18sion.
m1ss1onal7 tathers of ~e A.ugust1nian
vere Frei Sa turnino and
João. and 1 t wu their radio. opera t1ng through an &Da teur network wh1ch
wa.s wtrt1menta.1. 1n cODlllm1cat:iJ1g to the rest oi the world the pl1ght ai the
River po~tion.. tJJ1an.y IK"cmIPt1ng the PoH. Secre't.a17 to 8sk for help.
house bear1ng the bronze plate oí tJ18 ~p was pls,ced at our dispos~
~e8 0.1" lodgings ard work1ng quarters.
first hours o! our stq were spent gatherine; the usual oon~sed,
8u'bject.ive &:xl o~ contradicto1')" informatioo from relatives or the deceaaed,
e ee à.rxt voluntary collabora tors .
Santa Crus on the toll.owing day.
01 tl::e locali ty
vere made to vis! t
~brea 1s a amaD. town o.f' 2500 ~~1tanta l1ving 1n approximate~
OO.lBN o.t aU ducr!l)t1On8.
tX)Ols 18 tJ18 rule.
is no ~ter syste and tbe use ai' casa
water ia colleoted tram the river ~ the D8.jority' of the

Appendix II
p60ple, bIlt tilere 18 a group, l1v1ng &1f8.J' .trom the waterfront 'wb1ch avaUs
itself ot a 88epage at the back 01 tbe toIm. I t 18 d1tt1.cul t, UDder the
141
oirCUDStlno.., to decide wb1ch ~'SterI118 varRe. Blectrio 11ght t1'Om a generator
18 ava1lable trom 1900 W 2200 hrs..
':'he t,(nm 18 relative~ n~""'1 hJlviJ1g been i'ounded ~ the ~t!~-1e oi' ~
l&Bt oentu187- The reasCll .for 1t8 looation 18 obv1ous~ the fact tJ1at the
elevat1on of the river bank8 at tbat particuJ.ar place o.t.f'ers better guarantees
against the 'UlreatenjJ1g .fl.oOOs than at 8lJ1' other along the river. labrea is 'Ule
head ot the !tm1c1pio at tbe S8Jre D8M whi.ch CO\U1ted, at tJle last census made
in ~o, 211968 8 oul8 .
The ~-~- Ri ver
,
b ~, me of' tlw longest a.rfiuents of. the Amaza1, 18 chare.c~r1zed
bJ' a slow CtUTCt (aP1U"QXe one lmot and a Ialf), a:cept at t.ime3 01" tlood, that
18 t:rom ~ to Sept8ber. 'U1e period af' hea"Y' ra1nB-. Dur1ng ti1& t time , the
r1 vere,ides are oftc submerged. Tl18 _ters start receding 1n Septe1r.ber,uncoveI':1ng
8~p1ng eli>8DkMnte em beaches oO'ftred with sUt, which 'Ule 1nbabitarlt8 plt
't4 use for agJ8icuJ.'b1ral pJr}:X)89S.
.A. wll rooted Ca:18818U8 O.t Op1n1.~ amng old residente 18 tha t the 8fever"
USu&1q &ppea.r8 ifhm tbe watere 8tart to r18e, 1n AFil, ~ wbm th87 begjn
tA) num_, 1n September;
,
b l\Irã8 bas a. v~ w1IxJi!,~ course, w1 ~ D11Mrous loope wh1ch sometimes fora
contplete c~. W1'U11r1 tJlOse cirC\D\terel1Ces, th8re 18 often a sbO1"tcuttmg
#
chaz'.nel, caJled .Par8I3á', d1sS8CtJ.ng an ialalKi 88 i t jo1ns tt. main SU-eaJI'e lote
waãer S"w.w17 for 'tJ)e -seringue1ros- atn tbeir fAllrt1ies ~~!!p:1ng m 'tJ1e jungle.
li)st oi tbe JX)p1latim or 'U1e ltm1cip:io li" ml tlle river banka. Thq
bu.iJd -00_, tAtcll-covered 1mt8 00 stU te as a. precaut1on again8t the river' 8

Appendix II
-143 -
negafAive lar yeUow tever. OU- ot 78llow t.ver, bowever, 818t ba.ve been Im,lch
JOOre nunterous be.t'ore tJJe vacc1nation OampI.igns, me ot which took place rec8nt.1.l'
0-9ó3)~ ~ the o'ther hard" ~lar1a 18 stiU rampant" two cases vere deteoted
during tJ1e s~ of the oOl8aiss1mJ 011. of tJ)IBa had a beayY' ~ of falo1~
8ld d18d ,a t;ew houra &!'ter having been brought 1nO
It 18 tJ1ere.tore to be expeoted tJ1&t tba 1ntorme.tion be ratller miscellaneous
arrl contllsed.. Eve17 VP8 or rever 18 integrated 1nto a complC 8)1DdroE, 8D
abstract clinioal entiv doJKi.nated bJ' the ~st draat1o s,çtoms.. TtMi relat1ve~
spec .::,~ tea 'blr88 of 1GlloW fever are bown to 1n.tl.uenoe r8""" ~cences and
oolour tJ18 ovaraU picture 1n a speoial ~. Thia
afteots IU"inci~ ..
sympto:I'8' ~ obaracter of' the vta1 t am tbe colour of' tbe -k1Ji:. It 18 easy
to observe ~t those 8)'ÇtoJU 1ncrease 1n severiv as tJ1e 1nterrogation becoma
JOOl'e Fe8sing. St~ oontents beoa. darker axn darker w witb som 1nsi.'Stence
erd ".lp loo1dng like cor!ee grO\U1d8. The BalE happens vi tl1 .1&undice~ The normal,
01", at least usual, ~ coJaPlexi~ ot an anemia ch1ld of the brunet type w"ith
2~200.(xx) BHJ eas~ reaches the 8y8now.Yel~ !me. As to the .rever gobserva tions
a'e rareJz 8ub8tan~ated b)" aq th~ter readings and are de.f'iJ11te1l'
~ess1CX1i8~~
Tak1ng into acCO1mt tJ1ese factors of error for the bearsq iJlformat1an
#
on ~t eVmlts on tJ1e furiIS, 1t appears 'U1at at least s:SJ1oe 1.927 a severe
diaeaae attacking ~8tq tJ:. intantile }X)~tion 18 r~pant on botJ1 sides 0.1'
the river, fr~ as far up as Boca do Acre down to Can1~ and beyorde Si~ar
reporte, but vi thout 8n1' Jrecision, mention the J\lrC1á and the *deira as being
.1nvol"nd~ § ~-lies are decim teci. There ia an account .f'rom Paraz1nho (see -p)
~~--
§ Dr.. laoó AtAUah, Covemmsnt Doctor at Porto Velho (.Madeira) states t1J&t be ba8;
~ sem dia, during the four 19ars of h1s temIre ttmore t11aD a ~ed. ~~-Üt.irSl from
tJ1e sor1Dgais, brought to Porto -velho with 'U1e s~ symptoms as the Iabrea. CXl8S.
Thia m.torst4cm,
re tU1~ to Belam.
due to a chance encounter, was obt.ained after the ct:'!!!ll...issicnl s

Appendi:x: II
-144
where, 1n 1932, the settleIlBlt comprising 47 persons o.t' t11e same .t'am:iq was
reduced to JJ. i.~ one ~ntl1 (1Dfornsatl Sr~ Seba.s~ José J.hs10 de Paula,
Pt'OlltJt..o:.' ar Iabrea, who travelled. througb tbe placa at tbe time). Simi]---,.
evan~ occurred at p1.ac(;;g like Ig\t.aldads, 1n 1940" Peri 1n 19$0, ~anhão,
HL41t.ar!)..\ç.. .:\110-) ~' others. ~re recen~ (1963), an outburst 1n San IJ1i.s
CasSil.1J~i~ :!d.lled. eigbt childr9Il out Df. tan 0.1' tJ1e .~rim famU,:,r, and five ofuer
persons ai' ,dúch two \tere 22 and 2$ ~rs ald. In 1964 Eterial from febrilc,
iOWl"i~ p.'ltiel1ts \m& 1nocu.l9.ted i.."1to mtce, g~inea. pigs and ~~ celli wlth ne~ative
res1.1lts. Serolog1c~ studies revealed BI anti~ against A, B, a, Guas.
~"arIfA'era and Mebo-oomus Fever group arboviruseso The IX>PÜ4tion B~.oIed an
1.'tmm1v rate ar. cJ.ose to 8~ to :yell,:)w rever, presunabq due to tJ1e 1963
vac oL~ tion OaZ!ll»ign.
IlmInm1t,- teatB with a Tacaribe group antigen (AN 70';6.3)
31~.A.~ S lX>sitives aut ar 16 :1n tJ1e 1&4 and 113 dUutians amang oontacw, arA
4 out 0.1' 2') at 114 amaug nan~ntacts.
~~e 1965 outbre~
A ~.']sage t.1'~"1Sm1tted b1 t'rAe Radio Amãteo.Jr ÇOUP to which a miDaionary,
M
Frei Joao Teixeira, residing 1n Iabrea., belongs alort.ed tbe P.H. authorities
of. lWtaus who procoeded to request the 11elp of the leE.Co and tJ1is was the
rootive for the visit reported hers.
T'!le .,pioentre 01' the outbU1"st tl'lis yea:r rla3 toun..i to be '"-t Sarlta Cruz
de Pacia" a s~gal sitaated appro::jJrateq 15 miles down Tiver fl"om Iabrea.
two ar three miles frCXl1 tJ1e thec'lter 01' the 1.963 cvents. Two c~-1-1 ,irml o.f'
tJ'1e }'~lo .famiq and two others residjJlg ir. the D~ settle~nt had dicd, the
last one during tJ1e dolq su.ffered b)" t.'1e cODlnission in ~USo Sal. ta ~
v!sd. teci on the SUnday following the arrival at labrea. (October 17) with
'U"'.e object oí gathering M large an 8IOO\Ult oi people 88 poBsible j appro:'dJla te~
~ l'mndred per8ons were intel.v1e'..red. ~se were tlw survivillg cb11dren,
,

Appendix II
145 -
relat1vea, contacts, neighbours am arq R)re riv~ peOpl8 who had heard through
t1le gra{)S v"jJ18 tJ1&t a group o.t doctora had arrived. A8 usual 1n those cases
a lot oí' ~ bad to be devoted to out..pat1ct oliD1o work, a neo.~s&r1' step
towat"d bIDoc1 8aZ1q)le collect1an. 'n18 8&8 happened at nha da ~ÇL, ~ Il'lOtner
Y1s1t 2 mil.. up r1vC' t'rom Iabrea where the other case had. ooC\UTed. during the
f1rst week 0.1' Ootober.
In tot.m, there were tJ1ree dayS devoted to tl1e sarna work wi th. besides,
visita to patien~ aí ali kinds. People started arriving from as;
far as two daY8 distance and 1n.formation was tJ1Us gathered over a relative~
lar g e terr1 'to17 o No s't.o%'1' oí 8JV' suspicious case contracted 1n town was
recordedj tl18 sü1ent facts at tile o\lt-patient clinio vere tiIe two cases of ma.1.aria
at18 of which. heavi3:l' IX>Sitive for t~c1~. died :Ln a t.ew hauro.
Blocxi Samplss were taken at Santa Cruz, 1n town and at nha da Onça.
IIe.rq' b1o(Xl counts as tins penaitted were mdeJ testa ror uralbwll1ne were dona
ro"(1 ti11eJ;J'. FitV nine 01000-9 were ID:'eserved for serologicaJ. scre8nJJ1g and for
~'lidal teste
,
at Belem. Twel ve inocuJ.a tions 1n to baby mioo were dane wi th
material fran febrlle cases, and i11ne 1n Hip2 am GJ.I{ oelll.i-"1eso
~~ was a-tte.'!1pted at the nha da Onça. inside tJ1e houses and 1n tJ1e
nsigbõOl1r1ng bush. ~tomlOg:l.cal cap'blres were IDade .in the torest close to t.~
hO"l.1.Ses am at a }X)int a tew miles a~ !rom the river into the rain torest.
The l_qui ta. de Me~ f~
SpeC~ ~t1cm mst be made or tJ1i8 f~ for several rea,.,ons I
&) it BeeIr8 to have borne the brunt of the rever OIlSlaugbt tJ1is year together
-dtll tbe reJ.,ated Saboia de ~1o, living at a short distanoe from their seringaJ.J
b) the b1st,orr of this group 18 reJ:n'8sentative ar most ar the epidemiologic~
,
i1appenings one bears aí :Lu connection with tJle -febre Feta.. aí the ~, the
,
J1JrIIA azn tJ1e *deira as lar as peOpl8 oan r~r J c) ~y af..torded t.~e ool7
As

o. v.2-'1
Appendix II
Op}X)r'bm1ties fca- object.1" cl1n1cal observatim. dur:ing tJ1e course oi'
dis eaa 8 az1d d) tlJar provided aeveral tiver samplea. .trom biopsy atn
-146
a'.lto~3', which tcn-m tJ1e 0rl1J' base a~U~le
.1nto~ ti~.
todq for b1sto8}8tJ1ologicaJI.
The episode at Santa (h-uz st.D.rted 1n September witb tho deat1-..s ot ~ Q." .
chjJ.dren (Ra:1nm.lndo ntemat' Sa~cis. de ~lo, age 10, Sept.6-10" am Ra1JImx!o ma
de 1"1810, age 7, Sept. 18-23). Two teenagers followed by a fet-l da-ys (Franc.1.sco
de Assis ~squ1ta de ~10, age 17, Sept.27.ther !.-lo aOO the six surviv1ng children, ar which onJ.i" ~ia da
were isolated 1n a basement at the Hospital da Santa casae There, l-m-ia da
COnBolação took siok on November 6 and died on tile U'th'e
A l1ver sample f'rom ~ia Ra~da was taken at IAbrea by a m.1.ss1on~,
absent at tl1e t1D\e oí 'tJ1e vis1t, and having in h.is p888e881on a viscerotome
1eft over fraD the o1d Serv1ce. Th.1.s sample, together with another .tran tho
autop.:"}- a! luis. da Consolação arxl the b1o1::Eies made at the hospitaJ. on Carlos
*-ria das Graças and l'ária. Rita. oonstitute up to now the basis for a
tentative pathologicaJ. diagnosis.
'l'hay

Appendix II
-147 -
~k8 =. the epid81o1ca 01' ~ d.1.8eas!
.& In taote e8l"p trom the ...ot reporte, tale., leg8Dd8 â tantut1a
1nterpoet8t1ons. It 18 1IKlub1table that there has bec, 1.or ~ 18&%'8,
~
reJ1pnt epider1ic among tJj8 3qUAtters ard ser1nsueirCls 01. the PurUs r1vcr,
and
probabJ.y azoong thoS8 of" the
,
Juruã and ~e.ira rivers. The d1s eas e s t&1'1kes,
year arter year, attaCldng mos~ the famjJjeslnumerous broods with a reported.
predominance or males (not conf1rmed thio year) o In the absense 01: ar-~ oer..ous,
diagrJoet1c data and death declaratian, it 1s dirticult to assess quantitat1ve~
the case :IJ1cidence and the .rat..al1~ rate. Adul ta are no t eIempt a.lÜ an
occ~iona1 case occurs. ~:111e. the last five years, therc have bem outbursts
on a s~etch 01. appraxiDm teIJ' 40 ld.lometers I wi th labrea as the geograpr.icaJ.
centre (see map).
A most important f.eature is tho absence of ir~ections in tcW..lS. 1~O case
ws ever been re~rted aa hav:iJlg been contracted ~ Iabree.. proper, cr m
Porto Veli1o. Th1s is remarkable ()spec~ ir! view of Ulc tact that f.é.tients
are o.r~ brought to town 1n the aoute 8tage, aocompanied by oonta.otB possib~
.in the iJ1cubation st.'ige. It ia difricult to ascertaÚl at Ulis stage ir tJrls
15 due to some infectiou8 rcechani~ limited to tl1e jungle, 01' 11' it de,iJ8nds on
11.f'e condi t1ons . It S8eJ11S also tc. ellidnate the idea c.f a water-borne
1nfection, s1nCEI both the totm dwellers azn the squatters use the SB.n1e SUP~.
th3. t ~-(I the ~.
Another 1nterefJt1r.t; characteristic ia the a~rent fB.ll1fii81 involveD'.erJ.t.
TJ1EI :'~lD fami~ iD .t'a.r from 87'..ceptional. Tbe fallow1ng lis~ ar deceB,soo in
t\-1O fam1lies are representative of the mcxie 01' attack such as it 19 told .troa
aeveral lCIca.1i tiea jJ-. the past I
a.

-148-

Appendi:x: II
/lJ.mm f.~ -~C IA11z
Dea't11s in 1963
N~ Age
An ton1o Teles de 4~-1D1 10
l-iu-ia do Socorro 7
An~ :J.;1.~l('s 6
Franc1s~ 10
Ad8J.'bert.e 2
~ 1
Joee 17
ZaUJ.ce 2
N
SeXo .Da te
K S;m
r26~
H 29.IV
H S.,
M e.v
)1 9.VI
H~VI
F 23.VI
-149-
1-~ fe.r~ -~ta Crl1.z
Dea~ in 1.965
Neme Age SeX DatA
RR..~n1do n temaot. S.Melo 10 H 10.n
Raimndo ma 7 x 2~ .:tt
Francisco do Assis; 17 1-Á 2.x
1W1s. do Socorro 16 r 3ex
*ria r~, n F 18XI
*ria da Consolação .9 F lJ.éXI
(p1.us 4 casea, FesentJ:I-
1n convale8o~nee)
S~mJJ~.r ep18odes are reported from ~ more looalities alcr~ tJl.E: ri~lSr.
It. wst be ~inted out ~t# at, the sane tiIIE, other c~es occur 1n ne:1r,hbour1ng
W.J.r~e.! cr at short distances up or d~ rlver. It is possible t1.erefore t!'.e.t
tJle f~~l1n"'OlveMEmt ie due .):iJnp~ to }%'opmqu1ty am exposure to the ssmo
so-.:.rce 01" 1n.tecticn ratber ~ to som hereüitalT tra.it as happens, for instanCE'o,
itl .rt'!.vis~
~t08 to1oa
S)'JIJPtorE reported below are tbose x:a-esented by tJ1e ti-n *lo ch1ldren
wmse observation s tarted ir. labrea arxi ~ms .turtl1er nmde possible ~ the
tro~~lX>l4~t,1on to a Bel- hospital of tJ1e
, ,
whole ~. It ~t l.>e sajd tha.t
What 'fflE ob8erv~ cn tJ1Ose patients 18 gross~ 1n ag;;;~t with reporte frca
past cases aOO -7 be considered as reJE'esmt&.tive. ~ceptions to tbis sta.teIr.ent,
ho'We:-v'er, n.'.st be made 1n oolU'l.ection n'tJl two syçtom8, the jaundioe ard the
cba.ra,cter 01' 'tJ1e vomi t. 1-0 ~~ige- -or tbe five c~-11drm under observation,
m":J:;- on~ (*ria das Graças) showed definite jaur.d1.oe invclving the s1d.n and
1W.CO8ae;~ It lasted ~ for a tew days. The other8 had. the usual pale ~(;)Jnplex1m
ane..'!dc" tarasit.1zed, 1mdernourished,i tropical childrc of the bnmet 'typeJ
it 18 r8DarkabJ.e tJ1at the D>ther oonsidered ths as -,e11DwW and stated that tbe

Appendix II
-150 -
otJ1er cases back bCI~ had ~ bem 8yellow, jUBt l1ke 'Ula t..
11\ tJ1ree of.
cases observed at 3el('In; (Carlos Magno, Maria R1t.a, !'Ar1a do Rosario) a l1ght
sub-'1ot.erus not involv1ng the conjlmct1vae deft1Dl':ed d\1r:lnii the third week of
tile di~A:1S$~
In the acute, fatal ~e (~ia Consolação) wbo d1ed after five
~ ot illness, no j~~'~1C8 appeared.
a.. VCIa1 t -Scant)", dark vomi t wu ObBe..~vd
.
m1l7 1n the 088e 01' l-tu.'ia da Consolação, sbortJ3' before death. According to the
motJ1er' 8 op:1Jlion baaed CX'1 tbe experience ot her tour desd, 9.i'd tJ1e other
fataJ.i~~ at santa crus, 8whS1 thq vomit Llood, tJ187 di.-. ~ omours wi tJ1
observations b)r other vimesses suoh as Father Saturn1no who asserts: that nona
oi' t1lOse ~lO 8vnldted. bloodll surv1ved. 'l'he 8)'Iq)tom appeara t4 be l'B-tbognarKmic
of' V917 acute, fulluinltting Ja'OC8SSes; vith a f1'98 to 81% d~ durat.1cm, and 1s
absent in long ~, B1-1baC1.1te cases. 30 Fenr -A d1Bt1nction l1IlSt be cstE:.blished
betweSl the course of 'U1e rever in acute cases and the BU~cute, respectiveqo
The reporte b-CD tll*J. la7 peo{:ü.. are :1n &gre8Ellt witJ1 what was obeerved. at tJle
hospi ~. SlL~te cases shaw a subtebrUe st&t4 witll lIOrning remiss.1.ons
osc:i.llatirg betwem noraI and 38 o. at mst. farte p931"J! appear ooca.si~
during oon~scence. The ta~ case presented three peaks ot .39, .39.5
38 c~ w1 th 8)rnjJtg rem1ssions to nOl~. During tJX>ee ~J8 tJ1e ~se rats
de{)~[~~l 1'rom 80 to ~ 4. ~e8t1~ -~~ anorex1a,. nausea am voDt1. tir"1g
vere general. AbdominaJ. tenden'leS8 and Mteorism, oocasianal d1arrhea, 8~t1mea
aJ. tema t1ng vi th cone t1~ tion. No "01&1' swola"' obsen-oo-. I!!!!: -variOU8
degrees or enlargel8nt and tenderness.
In one o.t the oases:, coosiderable
8nla.rgEDnalt ~1d ~. 18 2 Spl8C1 developed dur1ng convalescence. 6. Nervous -SUb-
acute casos presmted some degree of ad1I2amia and de!4"e8sion. A CODIIK)11 prodr0m8
o.f the disease ia a lack of desire to p1q and rim aro1DMi; ~ child 18 quiet,
~ ead, p1X)tophobic and laohrJ1:1Ose.Ao certain ~egree or obnubnation develope.
111 the fatal case there 'Wa8 an earq se1ld.'l!Gtu~r SOCRl followed b7 com.

Appendix 11
151 -
A. sate of uc1tat1on witl1 feeble scr8atlr1.ng and apparmt distreso ocourred
m18 dq betore deatJ1 azn was tolloved 'bJ' relapse into COSe This a)'ndrome 18
1811t1oned 1n 8»st of tJ18 b18tor1es of fatal cases obta1ned at IAbrea.
7~ 0tJ18r8 -SU~]]s,. 8!1d posterior cervioal g8Zlgli& .free, enlargcd,
.
p8.1.DN~ 8~ ~-!.-~~t1M 1ng! -BO\1t1na ha8K)graZJm W8re started at Labrea
and cont.1-'1U.~ at Sel_. It mlJ8t be borne 1n miM wha 1nterFe~ resulte tbat
tJ1is W88 not a group of ~ chUdrc even betore the aotual d1sease started~
In 'bIo acore ar blood coun~ lDade &IOOng oontaou, an BBO count abave 2~OO.OOO
was lO\md to be exooptionaJio Càle ot tJlOS8 exceptions was *ria das Graças,
(with 4:.200~OOO) tha ~ one to develop definite jamx11oe~ I t ia di!. fioul t w
gauge the sign:Lf1cance 01' 'uilirubm teste with that backgrown. DuriJ1g the
disease, there appears to be ao. earq loucopenia with relativa l1DLphoaytosis, a
condition vh1ah reverses i~e1.t af'ter the second weeko Eosino,l:i111.ia ia present I
btlt here, aga1n. aJ.l tJ1e ~~~'rom1 ver. 8hown to be with a \"aJ.'ied talUla o!
1ntes~l ~ite8. As oould be c:pected, organio iron io verr low, ~glob1n
belaw nol~ coaguJ.at1cm t1l18 rrolonged, sediRmtation rate h1gh. Ur1naJ3Sis
soowed. t2l8 }rOSC1C8 of. aJ.bum1n am casta 1n tn8 subacute cases. GluOOS8 wa.a
positive m tbe Carloe *gno case.
raiver fImot1on tes te
m.sc~pboretio }roteioogram, van der Bez'gh, transam1nBBe, l1pid and
oholesteroJ. dosage 1Mr8 pertor.-i at the hosp1~ on an pa'tients-. The resulta
at tJ1Ose teste urdra~ 1ndioate a severa damage ar the tiver parenO1V'JJae
~gica1 tes g,
Inooula t1ona 1nto 1n.tant wh1 te .mice, guinea pig5 and tissue cul 'b11"e mbes
wi't11 blood, urine, biol:B7 and necroPS7 ateria]. o.t' aCllte and subacute cases were
not

Appendix II
\m1.torm1:3'
L
negat.1ve.
-152 -
~~log1~1 RI eC1'8ening aga1nst:a, '81, li1cambo, *1&rOI P1'XIm8.,
Yell~1 rever, Bus~J.ara, nh8UB, SLE, BuJa,nÍ and Iooarac! lave the tollow1ng
resulte a
:8
HJc..*7.
~'
PJxo
U'
aJa.
1m.
sm
100Buj;
1964 (ltIF) 1965 (October)
o
1
9
O
O
i
u 312
~r_iolog1.cal 1Dves~ia tiOD8
o
2
12 O
O
4
3
16
3
1
1
W8re pt)rformod at the Iaborator)" or. the Evandro C2'1agas Institute.
Standard cultur1ng a'ttGnpts in isolatian ~ia using blooo, urine" !ecoo"
(oliw:t.:lOU ~ dejec~ a.nci b"i swab), 1ntest:1naJL DIloosa- bile- were an
negative for ~l,~~~11~. ~e~ ard Isptospira~
'l118 WidaJ. testa:
~1i'-.ane sera were tested .rrom suspicioUB cases, contacts etc. .tJ.'om
Iabrc". and Santa Cl~. These sera were taken .1Jl Octoller d1J.1"~ the V'"ls1t, r-Dd
3~ o! ~ belonged to the children w.1O later took siak and vere brought
,
to Bel8!1. It 'RB tJlUS possible to aq.ke a few paired teste. 'n-.e all rOU1Ü
resuJ. t or tJ18 tests is &os t.oll~.
NQ of. peoplc wbo8e serUJU was tested
~1t1ve3 to & ro m
TO 'l'H .1.(11)
TH B
Doubt-t'u.1 ~O 1/dO)
-(lIJo 1/80 )
2l
.3
3:
1
3
1
NegaUve to an antigens 10

11
-153 -
Four of the pos1t1ves are .rram the pat1ents.
~M. are listed b)1' cãsel
H~ ar patient
~-~~ .u
l'Ook8~àk., date unce t,aj.n bu t
de.t:illi.~ in.forstion an a "tever8
ar several da)'88 before death af
brot1'lers whioh ba.ppened Ia. te Sept.
The -;resent candi~ ia eit.~ a.
relapse ar a d1fter~t entit7.
~ ~~:.8 15
-~ 81cà" da 'te tmOertain,
was de.t1ni tel1' jl.\mdiced.
Carlos ~.. oS
-~s~ ~:1ng tJ1e second
weak of. OctA>ber.
!-'l3.ri.~ do Rosario .,# ~m~ 8 29.
!õõi s1'~
-
~iA da~$ção:r.9 ~ov~
'l'OOK sic1t NOV8IDber ~
~_~gioaJ.firxi1n gs
Convers ions or ti ter
Da.'t8 Da~ Result
of ot
oolleotion test 'l'O m .f!.~
~~- .~ ~:-
17~ 3~ Jn,60 1/'20 O
6.u 8.'II 1/).60 1/)20 O
l2.n 27.u ~ 1/320 O
2~ 8~ ~ 1/80 o o
17.'1
6.u
11.x
2.xI
~
17J
6.xI
6.u
3.u ~o
8 OU lido
1/80
1/80
1.7.n 1/20 1/20
8 fXI lIdO1.7.xx ~O
1/40 1/20
17.u
8en
17.u
~
~g
1/40
,
Two 11ver samp1.ee Obta1l1ed- one at Iabreac and the other at Belem,
o
1/640
J/)fJo
J/~O
]/320
1/;1.60
~~
esW)l:Lah the presence. b"OJa two CMeS with lesa tJ1an aeven days evolution, 0.1"
un1form a.rn dissEinated pareno~ destructian witJ1 innamnatOl7 per1lX>rta1.
.inr:Utrat1on or. the D>nonuolear type.
SeveraI biopsies were performed at 'lhe hospi~, but, unfortU&"18.t"3l3',
onJ;J' one reached the pa."tI1ologic~ laboratol")" at the Institute. I t 18 .fram t.he
o oo
o
lfto
o
o
o
o o

Appendi:z: 11
-154-
case *ria Ria (see above), 1u ap!Pt"ox:I.matel:l' tlle third week ot disease and
apparent convalescence.
It tn"esents tlle piCUlre or. a regenera tion ~
genera.li39i necrosis, witb den3e porta.l a:n 1ntralobular in.t.iJ.tra'Uon, mn
endotJwJ..:7.~ ,1n'Olitera.'tia'l.j there ia also an 1noeptive fibrosis a.nd com
d1ss~ted r8lm13.n~ ai' neorosis.
D_ia~O3_~
Iellow Fever I can be disoarded on ol:1nical. and h1sto~thologio."3Jl.
__8'
grow1d.s. 0tJler reasons I sever3J. oi' the disoased ch:Udren bad been vaocinated
in tl16 J.96J oalI;)a.1gnj although th8 jungle ~1low rever vedror ~~-e
~&~~ spegazz.in1 was found in the vicinit'1' or tJ1e houses it was 8:':tr~l-v' -
scarse (l s~ciJIIm par tllree man-hours at Santa c:-uz) due to seaaona.I oondition3.
Brolcella: a possibility which was considered because ot 'Ule propinguity
oí pigs~ Disoarded on bacteriologicü and clinica.1. grow1ds.
~'tta1oa~1 1noo~tible with type 01' rever a.ld other negative
clinical aspec-w.
Negative 1no~tions. Same for Leptosp1rosis.
~1d ~ ..E!~1d ~P' The resul t.9 or the W:1.dü teste seemed,
at a certain stage to have SOlE signifioance. Th1.s was grea~ djJmood b)" the
totaJ. fail.\1r8 ar. aJJ. tl1e attempto at culturir1&. Also, certain cUn1c4
s~toM such as tJ1e temperature did not evolve typiCa1q, even m tJ1e acute
Tl8 1nteI"pretation aí the behaviour aí tb8 agg1.u+.i~iDS rera.iJls d1fficult,
1.D1less one ia reaà7 to accept the }X>ssibUi 'tV" af a nCXl~pecific anaDt8StiC reacticm
lU'a1IPt.ed 1:7 an 1nter~ant 1n:tection. Even 80, 1t appears ~~t an endemic
#
of. the ~1d~a~1d groUp exista along tJ:18 Purt:1S. I wha t exten t tJds
can 1nnuence the r1vtlm1 and characteristics ar another 1n.tect1on remains to be
seen.

Appendix II
~~-' ~~~~
-155 -
,
For a o~ time alre8i17 the "b1ack f.everlt 01 the I\1rús r1ver ha8 bem
assoo1ated witb a fom of epidemo hepat1t18. A. f.ew 11 ver BampleS, triokl1rJi
to Rio de .II.n81ro tJ1rough tJ1e Serviço de ~emiu lh1ra1s 1n pat years were
D>St11' d:.Lagnosed as "negativo para Febre Amare~., but 1n 1963, a d:LagnOOis oi
~ he~titisn 'TJas IDade by Dr~ }oãrio de lmaes, p9.tJ1Qlogist or the Instituto
Nacicnla.l de Pesquisas da AD8zônia, from Saç1.8S obtained by a medic~ group
SSlt. to T;1brea; 'bJ' t.h8 l\1blic Health Secre't.a17 oí A-.sonu (Iilnaus).
The "':l.1.D.icaJ. obaervaUons
,
IDade at Iabrea ando at the Belem. hospital lU'Ovide
the first object1w data cm. the 8)'ÇtaIE wbich characterize the acute and the
subaCl1te cases.
Th87 g:1.ve also the first labora tO17 tests evar made. The
auto~y .?.~ bio:P87 _teriaJL are also the first which oan be related to obaerved
cas e3.
Th8 a11nicaJ. teatures. 'Ule laboratory resulta and fin~-Jr'. the pathol.ogica1.
.t"1nd:iJ1gs concur in establishing the existenoe 01' an acute disease severe~
1nvolving the l.1ver ~o1vmso The process 18 somtimes slow and subacute.
1asting 8ever~ weeks, at other times .t\lJ'IIrlnAting and ending 1n death after
fi w '00 s j.j(". dqs . 'l'he etiolog)" of the disease is a mtter for conjectureJ
ti1e ~th9S1s of. a vjrus origin ia curr8ntq tavored azn 18 supported 'b7 ti18
total negativ1~ of al]L ~ att8Pt8 at 1Do~t1ng or culturing~ Plana are
being ~e to tt'3' the Detro1 t-6 t1sSU8 cul 'b1re call line. which 15 not ava1lable
at the Jn'esent t.1Jrs;
j; tea"bÃre of. tbe disease whiob W8B misleadiDg at fust 1n tll8 interprets.tion
oí tlle syhl~ was the exception~ h1gh mrta.l1ty rate 8lIK)ng chiJ.rlren.
SUch a Dl)rtal1t," rai5es ~ al ternative of a new viros strain ot exceptiona1

Appendix II
-156 -
v1rQlence on tll& one bazn, or of a. 1a1O1m agent operating on a spec1~
1iW81o1ogical terrC.n ot particular de.tic1SlOJ" on th8 other~ The 8olut1on
of' tJ}js quection 1fOuld require lIIloh DDre observat.ion, date. m1d ~. teriBJ. than
tllSot wb1ch 18 ava~ble todq~
CalClua1ons
Fram daw obtã.ined in the course of a vis1t to !B.brea by a colmTl1&sion
from tlle Ir1St.1tuto Ev'c1l1dro (21agas, 8I1d from obse:rvations Dfade later at the
bo8p1~ or Belc (santa. Casa;) oi' S cases brought from Is.brea, 1t 1s jX)8B1ble
to confirm conclusions rea.ched by prev10U8 investigators in 1963 and ~6J~ aa
tc tJ"'..e Dature o:' the d1s~se. The evidence" especi~ the his~tJ1o1og1caJ.
er...dence. seem to !X)int w an cI::d.dEic torm o:f: a ~ hepat1t1s.
li)wever,
because of certa1n teatures, especiéllJ3 the high JaOrtalit," rate, it 1s felt
tbat dif'.terent l'B"Ocesses are at work which would juat1i)' ~t1}P.r research.
Ecologic~, tJ1ere is alBo a disturbjrJg observation~ aJ.read7 rccorded by
the JNPA ~sion 1n 1963. coní'irmd by recent 1nteITogation .1n I.a.brea and
re!X)ru ~ Porto Vel})O & no attack has ever occurred 1n to1m~ Th8 disease
a1'.teots ~8~ tJle ohjJ.drm" wi 'th an excepti~ adul t, .trom t1le settlel8l~
a.lcrng tile rivers,; i8 this due to condit1o11S of. l1.fe in gener~, or to s~
jn.t'ectious 1'aotor lmkoo to tJle fo~st1 'l'he answer to tb1e ques Um cannot
be I~e at thi8 ste.ge. A In'olonced "tudy -m sitUo" i8 ~b2.bl.;'" neoess~
to tl1rotf li:oCire ligbt on t1'd8 Oa'1pUcated endoecpidetno sitAlation. Frcml
scientific, economic e.:1d 1mrnD.nit.e.riM considerations, it 15 felt tl12.t such ao
St\I:J:r would be jU8tif1ed, 1n spite oi' aJJ. th8 di.fficulties ard obsta.clea 1nvol~

APPENDIX III
VIRUS R&5EARCH IN AMAZONIA
This review covers virus isolations and epidemiological studies
carried out in the Amazon basin, which 1ies main1y in Brazi1 but a1so
includee parte of Colombia, Ecuador, Peru and Bolivia drained by affluente
of the Amazon r1ver.
The viruses found in this area fall intp 2 broad
groups: those which are the scourge of mankind and bis domestic animals
the world over, and those which are peculiar to the Amazon forest and its
creatures, transmitted by mosquitoes and only infecting man sporadically
when he ventures to cut down the forest to build a road, a homestead, or
to cultivate crops.
The first group has been relatively well studied in
other parts of the world, and the viruses in it can be classified on the
basis of their structure and biochemical properties. A recently proposed
classificationl is used in this review, which discusses only those viruses
which have actually been isolated in Amazonia, or for which there are sero-
logical data. Many common virus diseases such as chickenpox, rubella and
infectious hepatitis are of course present in the region, but are not men-
tioned for lack of virological data. The second group is less well underare
stood, and structural and biochemical data 4e/scanty, so that the viruses
are classified on the basis of their biological properties. Most of these
appear to be arthropod-borne animal viruses (arboviruses), and can be sub-
2
divided luto groups according to their serological relationships .
Actual isolation of viruses from Amazonia began relatively recently
Ye11ow fever had been known in the region for hundreds of years, but the
first iso1ation of the virus was made on1y in 1954, by the staff of the
3
newly-established Belem Virus Laboratory . In 1955, scientists from the
USA isolated strains of an arbovirus from Okinawan colonists on the Bolivian
4
Amazon , and collected human gera from the Peruvian and Bolivian Amazon for

Beni, Brazil. Highways,
Appendix III
158 -
5
yellow fever protection tests and tests for other arbovirus antibodies .
More recently, a Comisión de Investigación de la Fiebre Hemorrágica del
with support from the U.S. National Institutes of Health, has been
investigating a highly fatal virus epidemic in the Bolivian Amazon6. But
the major effort in virus isolation, identification and serological survey
in Amazonia has been made by workers at the Instituto Evandro Chagas, Belem,
Para State
At that institute the Belem Virus Laboratory (BVL) was
estab1ished in 1954 by an agreement between the Funda,ão Serviço Especial
de Saúde Pública of Brazil and the Rockefeller Foundation. For more than
9 years this laboratory was under the inspired direction of Dr. O.R. Causey
of the Rockefeller Foundation, ably assisted by bis wife. In 1956 the agree-
ment was renewed and the Department of Microbiology of the University of
Brazil, Rio de Janeiro, joined the signatories. Since then further collabo-
ration has come from the National Research Council and National Museum of
Brasil, the Instituto Oswaldo Cruz in Rio de Janeiro, the Instituto de
Pesquisas e Experimentação Agropecuárias do Norte and the Water Department
at Belem, the Department of Rural Endemics, the National Department of
and Indústria e Comércio de Minérios S/A in Amapá Territory. The
7
organization of the laboratory has been described by Causey . Between
November 1954 and the end of 1965, over 2000 iso1ations of at least 60
different virus types had been made at the BVL, and some 40 of these were
originally discovered by that laboratory. Twentynine af thase types are as
yet known only from Amazonia.
In 1961 a tissue cu1ture 1aboratory was a1so estab1ished at the Instituto
Evandro Chagas, with the help oí a grant from the Rockefeller Foundation, and
this is now directed by Dr. F.P. Pinheiro. Before it was even fully opera-
tional it was pressed into service to diagnose a poliomyelitis epidemic in

Appendix III
159
and other enterovirus and arbovirus studies have been carried out
there, the details af which will be published elsewhere.
The laboratory of the Departamento de Defesa Sanitária Animal in Belem
has been active in diagnosing cases of rabies in Amazonia, and other labora-
tories outside the region, such as the Pan-American Foot-and-Mouth Disease
Center in Rio de Janeiro, have isolated veterinary viruses from material
sent from the Amazon.
Variola
Herpes
POXVIRIDAE
A sing1e iso1ation was made in 1961 at the BVL, from a Be1em patient9.
ADENOVIRIDAE
Types 2,5,7 and 11 have been iSo1ated from Be1em patients10
HERPESVIRIDAE
Two strains were iso1ated at BVL in 1960 froro oral aphthae of chi1dren,
and were identified by CF testing using hyperimmune herpes serum supplied
by the Trinidad Regional Virus Laboratory. strains have been isolat-
ed annually in mice and tissue culture from Belem patientslO,61 Of 3 i80-
1ations made in 1965 from chi1dren aged 3 to 12 years with stomatitis, one
c ame from a child with deliriumlO The virus has been recovered from stool
specimens as well as oral swabs. In gera from the Peruvian Amazon, comple-
5
ment-fixing antibody rates rise to 97% with age

Appendix III
Influenza
-160 -
MYXOVIRIDAE
The 1957 pandemic of the Asian A virus affected Be1em, where severa1
strains were isolated between September and Decemberll
Mumps
A serological survey among inhabitants of the Peruvian Amazon revealed
75% positive by complement fixation test5
Ne~cas_t_l_e- _disease-
Nine strains iso1ated from sick chickens from Be1em in 1958 were identi-
fied by the haemagglutination inhibition test, and 2 further strains were
obtained in 1959 and 2 in 1960 in Be1em9. Poultry farmers in Pará State
routinely vaccinate their flocks against the disease
Rabies
A few strains are isolated every year at BVL f10m suspect animal
material submitted. In 1959 two strains were iso1ated from dogs and one
each from a horse, a goat and a cow, whi1st in 1960 three more strains were
isolated from Belem dogs9. In 1965 one strain was iso1ated from a dog's
brain from Belem. Rabies is commonly diagnosed by the Faraco test at the
laboratories of the Departamento de Defesa Sanitária Animal in Belem, and
they have recorded bat rabies from Marabá, São Miguel do Guamá, and Altamira
in Pará State over the past few years, with another isolate from a pool of
10 brains of vampire bats, taken in Ju1y 1965 at Parintins, Amazonas State.
Cocal viru~
STOMATOVIRIDAE
This is morphologically and serologically close to Indiana vesicular

sera8.
Appendix III
161 -
stomatitis virus, so is classed here. The prototype was iso1ated in 1961
Mites of the garoe genus taken from a rat of the garoe genus trapped 92 km.
from Be1em on the Be1em-Brasi1ia highway in 1962 yielded another strain12
Poliovi!us
NAPOVIRIDAE
AlI 3 types are eudemic iu Pará, and presumably throughout Amazonia,
Between October 1961 and March 1962 an epidemic occurred in Be1em, during
which 27 type 1, two type 2 and two type 3 po1ioviruses were iso1ated from
faeces and rectal swabs, and homologous antibodies were found in the patients'
Later in 1962 three more strains of type 1 were iso1ated, and in
~-_. _1n
196'4 two more of type 1 and one each of types 2 and 3--. ln 1965 type 1 was
iso1ated from 2 para1ysed chi1dren aged 1 1/2 and 3 years at Be1em10.
Neutra1izing antibody to a11 3 types was found in 1955 in gera from the
5
Peruvian and Bo1ivian Amazon. The commonest antibody among chi1dren under 5
years old was type 1 in the Bolivian sample and type 3 in the Peruvian sample.
Coxsackievirus
An untyped Coxsackievirus was isolated from a 20-month-old child, one
of the patients who a1so produced po1io type 1 in the 1961-2 Be1em po1ioepidemic8.
In 1963 a strain of B4 and in 1964 one of AS were iso1ated10
In 1965 eight iso1ates of Coxsackie A viruses (awaiting typing) were made
at Belem from children aged 1-7 years, with symptoms ranging from nil through
pyrexia to paralysis; a strain of A2 was isolated from the paralysed child
aged 1 1/2 years who a1so produced po1io 1, one strain of A4 and 2 of AS
were obtained from 1-2 year o1d chi1dren with pyrexia, and 3 strains of B2

odents, Lingual 1956,
Appendix III
162
from ch11dren and an adult, aged 40, w1th symptoms of pleurod}~1a, or
diarrhoea and vomitinglO,
Enceohalomvocarditis virus
14
An ep1zoot1c occurred from January-Apr111960 1n Para State V1rus
wa8 isolated from 33 .ource.: wild rodent., opossum, bor.es, birds, 8entinel
mice and mosquitoes.
Ant1bod1es were demonstrated 1n the blood of w11d
horse and cow. This virus has a1so been is01ated from mosquitoes
15
in Africa but transmission is thought to be genera11y by contact rather
c---
than by mosquito bite. It seems d!~f!ic~!~~however, to explain the sentinel
mouse infections other than by mosquito transmission.
Foot and mouth disease virus.--
This disease is common among livestock in Amazonia.
epithelium
from cases occurring in the region has been sent to laboratories in Recife
and Rio de Janeiro, which have isolated 2 strains of type O and one type A
in 1962 from Municipio Careiro, Manaus, and one type O from Amazonas State,
16
and one type C Rezende from Be1em in 1965 .A strain of type O was a1so
iso1ated in 1965 at the BVL from the b1ood of a Be1em bovine
Mouse poliovirus
Type GD7 has been isolated many times from the BVL mouse colony since
and appears to be endemic to the co1ony. It 1s most often p1cked up
when infant mice inoculated with material for virus isolation attempts are
kept beyond the usual observation period of 12-14 days.
THE ARBOVIRUSES
We depart from the structural classification heret because not enough
information is yet available on the ultrastructure of these viruses to
ensure that they forro a homogeneous group. Yet biologically the arthropod-
61

Appendix III
-163 -
borne animal viruses share the properties of transmission to vertebrates
by the bite of a haematophagous arthropod after a period of multiplication
within the arthropod, pathogenicity for mice on inoculation, and sensitivity
to ether, chloroform and sodium desoxycholate.
The arboviruses (and some other agente) isolated at the BVL are listed
in Tab1e 1, which a1so shows the sources frorn which each carne.
Fifteen of
them have been isolated from natural infections of man, and clinical pictures
for some of these are described by Causey ~~.l7. The results of human
serum surveys for arbovirus antibodies in the Amazon va11ey have been pub-
18 19 41 20
1ished ' , .Reservoirs and vectors are discussed by Causey .The
methods used to iso1ate arboviruses have been described e1sewhere17,21
21 17
together with summaries of the resu1ts obtained up to 1957 and May 1959
17 22-25
and so have the methods used for identification' -Further brief
26,27. ,summaries
of iso1ations up to Ju1y 1961 are given by Causey This
section updates these summaries to the end of 1965, inc1uding viro1ogica1
data from a mammal recapture programme in the Utinga forest, Belem28-30
Tab1es 2-10 summarize the iso1ations and resu1ts of serum surveys.
Maximum rather than average antibody rates are shown, in arder to give an
idea of the attack rate in a situation of active transmission. Thus human
rates are quoted for the locality with the highest incidence, and mammal
rates are given for the season when the rate is highest.
This system should
reveal the mammalian host in an epizootic situationt but the reservoir
during a quiescent period could of course be quite a different species.
Percentages are only quoted on samples of more than 50.
The arboviruses isolated in Amazonia are discussed below according to
serological group. The abbreviation HI is used for haemagglutination

us31, hibition, ver, ucambo. n,
Appendix III
-164-
N for neutralization, and CF for complement fixation.
antibodies should not be taken as definitive unless there is confirmatory
N test evidence.
Virus names given in parentheses have not yet been published, and
their mention here does not constitute a description.
GROUP A
The 7 virus types of this group so far isolated at the BVL include
Aura and Pixuna, which have not been isolated anywhere else. Muc amb o and
Pixuna form a sub-group with Venezuelan equine encephalomyelitis (VEE)
but VEE has not been confirmed in Brazil. Aura and Una have been
isolated in Brazil only from mosquitoes. Eastern and Western equine en-
cephalomyelitis viruses (EEE, WEE) although found in Amazonia have never
been isolated from mau there, in contrast to the situation in North America
Muc amb o (BeM 8)
The prototype was neutralized by a VEE antiserum and was identified as
a strain of VEE17,32. Subsequent CF, HI and cross-N testing showed that
although it i8 closely related to both VEE and Pixuna ViruSe8 and produced
1eukopaenia and viraemia on inocu1ation into a horse, it is a distinct
r 31 Two similar strains from Brazi1 so far tested have turned out to be
type
Mucambo virus has been iso1ated from a11 4 types of source (man,
wild and sentinel animais, and arthropods). There have been 6 isolations
f"' 1/ í: ,,---
froro man and 66 froro sentinel mice and monkeys; one monkey died after infec-
and in another from which EEE had been isolated a week earlier, there
was no Mucambo antibody formation, suggesting that interference might haveoccurred9.
Orvzomvs could well be the forest reservoir host, since it has
RI

.
Appendix III
-165
a higher titre viraemia than other forest mammals during infection with
this virus, and 13/21 of the iso1ations from wi1d animaIs carne from rats
of this genus. One of these was from haemorrhagic urine of an Oryzomys.
There was a1so one is01ation from a sentine1 white 1aboratory rat, together
with a matching isolation from the mosquitoes caught resting in the clay
9
pipe in which the rat was exposed. There was a single isolation from a
forest bird (~erythrocepha1a) in 1965. Mosquito iso1ates total 14
*
from Culex (including 8 from Culex B9 ), only 7 from other genera.
RI antibody rates of up to 34% in man and up to 43% in rodents and
some RI positive marsupiaIs have been found in the BraziIian Amazonl3,29
but conf1rmatory N test data 1s lack1ng. Neutralizing antibody has been
found in a vulture (Cora~yps) and a bat (Carollia perspicillata). There 15
evidence of cyclical activity of Mucambo virus in the Belem area, wit.h peaks
in alternate years.
Laboratory mice infected with Mucambo virus have encephalitis and
fatty changes in the liver
Pixuna (BeAr 35645)
80
This virus was described by Shope ~~.31 who demonstrated its close
relationship to VEE and Mucambo viruses and summarized the information on
the 3 strains known (2 from mosquitoes, one from Proechimvs). The same
paper recorda that a horse inoculated with the virus became immune, and
resisted challenge with Mucarnbo virus. No further isolations have been made,
except for one from mice inoculated with the blood of a sick laboratory
However, this could not be reisolated, and the patient showed no
* BVL code for a taxonomically difficult species, probably c. (Melanoconion)
portesiSO.

oasta1 aboratory p1dem1cs, . mazonia, entinel
Appendix III
-166 -
9
rise in antibody during convalescence .
mice infected with
Pixuna virus have encephalitis and sometimes focal hepatic necrosis80,
Eastern eauine encenhalomvelitis
This virus (Belem prototype BeAn 7526), which in the USA causes serious
and ep1zoot1cs among hor.es and pheasants, 1s frequent 1n mosqu1.-
toes and sentinel animal. in some years in the Belem area, and ap~arently
absent in others33. It has been isolated there in every month of the year
except November.
sick equines.
Causey ~ !!.34 describe an epizootic among horses in the
region of Para State (Bragan~a) in 1960, with 2 iso1ations from
There have been 20 iso1ations from Cu1ex (inc1uding 12 from
(~elanoconion) taeniopus) from the Belem forests and 3 from ~.taenior-
hynchus from the Bragança region.
One of the positive pools of Culex spp.,
the pool of Ce (Me) ~pissipes, the 3 pools of Culex B9 and 4 of those of
C.(M.) taeniopus contained mosquitoes captured on sentinel animaIs subsequent-
ly shown to have been circulating EEE virus, and therefore those isolations
should probably be discounted35 No human cases have been recorded from
but there was an unexp1ained high incidence of N antibody (12/41:
in 1953 in the popu1ation of Cameta, Para State.
9 13
that town was sti1112/55 over 6 years 1ater '
The RI antibody rate in
Isolates from the viscera
birds were not confirmed by reisolation, but 3% of Amazon forest birds
captured in 1963 were found to have RI antibody (the positives confirmed
by NT), whereas none of 610 rodents and on1y 1/148 marsupia1s captured in
the same area during the garoe season as the birds was RI positive, suggesting
35 34
a bird reservoir for the virus .In other years 3/19 Jacana and 12/126
Co1umbi~a11ina birds, and up to 5% of marsupia1s (Marmosa, Dide1phis,
13
Metachirops) and Proechimvs rodents have been found HI positive , but no
N tests were done.
chickens gave one isolation and 5 HI antibody

~.~(..;
Appendix III
167 -
35
conversions in February and Apri1 1963,2 conversions in Ju1y 1964 , and
a second iso1ation in December 1965.
Laboratory mice naturally infected with EEE have encephalitis and
80
lesions of the connective tissues .
Western eQu1ne enceohalomve11t1s
This virus has a lower case mortality in man in the USA than EEE, but
is a1so responsib1e for 1arge epidemics in man and horses there. It was
iso1ated for the first time in Amazonia in 1964 (BeAn 70010) from a bird
(Myrmotheru1a hauxwe11i) , and HI antibody rates of up to 14% (confirmed
by N testing) were found only in forest-dwelling birds, not in open fieId
35
birds nor in forest rodents or marsupiaIs. Only 3/80 human gera from
Para State have been found protective 19o
No human or equine cases have been
recorded from the Amazono,
,. H -1f
t~~'" ~ R- -~q
(2..(..J; ~.4
(2;; ~~f.)2
I ~,;::tL,~t~~
~~, I~... II
~G (tA .-.2 (/ ~... t f í..,..a~~ )
Mayaro
y2.
Six strains (BeIem prototype BeH 407) were isolated from patients
during an epidemic of fever among workers on the Rio Guama, Para State36,37
A further 6 cases, including one presumed laboratory infection, are recorded
at the BVL9. One of these was isolated in parallel in infant mice and
BHK-21 cel1 culturelO Neutra1izing antibody rates averaged 10% throughout
the Brazilian Amazon19,36 HI antibody rates are high among certain populations,
such as that of Tefe, on the upper Amazon river,
.
with a rate in 1960
of 60%13. Two iso1ations from 1izards are technica11y suspect; 25 iso1ations
/' 2 "c, }v.L1~~l. ~ , .{c ~ .si!'
from Haemagogus spp., compared with on1y 3 from mosquitoes of other species,
9
suggest primate involvement in the epidemiology . There have been no 1801a-
tions from wild vertebrates. HI antibody rates of 33% in monkeys, 7% in
rodents, 5% in marsupia1s, 28% in birds (Co1umbi~a11ina) and in 1/7 sloths

Appendix III
-168 -
(Bradypus) from the Amazon forest have been found13
, 1eukopaenia,
but no N tests have
been dane. There has been one isolation from a pool of mites (Gamasidae)
9
combed from 4 Orvzomvs rats .
An experimental cebus inoculated subcutaneously with Mayaro virus had
viraemia on post-inocu1ation days 4 & 6, but not 2,9 & 11,-
9
and produced high titre N antibodies. Laboratory mice infected with
80
Mayaro virus have encepha1itis and 1esions of the connective tissues .
4 ~
Uruma virus, iso1ated by Schaeffer ~ ~. from j patients during an
epidemic of fever among Okinawan immigrants in the Bolivian Amazon, is
38 39
considered by some to be a strain of Mayaro virus .Schmidt ~ ~.
discuss the serology of Uruma virus. Some 38% of the indigenous inhabitants
4
of that part of Bolivia had N antibodies .
Aura (BeAr 10315)
40
This was described by Causey ~ !l. It has been isolated only from
mosquitoes (~. serratus 5 strains, Culex (Melanoconion) spp. 1 strain).
HI antibody rates in man and animaIs in the BraziIian Amazon average Iess
«40 41
than 1% ' .Laboratory mice infected with Aura virus have encephaIitis
and myocardial degeneration
Una (BeAr 13136)
Causey !!. 2!.
80
40 a1so described this type from mosquitoes, and 10/12
of the isolates were from Psorophora spp. They found HI antibody rates of
3% in man, the RI positives being confirmed by N testing, and 1% or lesa in
rodents and domestic ungu1ates, in the Brazi1ian Amazon.
41
rate of on1y 0.8% was found in the Municipio of Be1em .
A human HI antibody

Appendix III
-169
GROUP B
Only 4 virus types of this group have been isolated in Amazonia,
although there 18 serolog1cal ev1dence that a f1fth type, dengue, may have
been present some years 8g0.
This is based on a 1953 serum survey which
showed that (a) sera protective against dengue 1 were commoner than sera
protective against dengue 2, and (b) no person lesa than 25 years old had
dengue protective antibody, this probab1y being re1ated to the eradication
19
of the vector, Aedes ae~ypti, from the region within that period .The
4 types which have been isolated are yellow rever, St. Louis encephalitis
(SLE) , 11heus and Bussuquara viruses.
Yellow fever
Diagnosis of this disease had been made for many years in Amazonia
on the basis of the pathology of liver biopsy specimens secured by an
efficient viscerotomy service in Brazil. Vaccinations have been carried
out alI over the region. 521.000 in the years 1937-54 in the Brazi1ian
42
Amazon a1one .The N antibody rate for an unvaccinated area (Capim River)
19
was found to be very c1ose to that for adu1ts from alI areas combined .
The first iso1ations of the virus in Amazonia were made at Be1em
(prototype BeH 111) from 21 human cases and 16 ~osquito poo1s, between
November 1954 and August 1955.
I'
The hurnan cases carne frorn 2 epidernics near
Belem, one in labourers opening a road through the Oriboca forest, the other
3 43
in adults and children of a charcoal-burning and farming community at Apeu '
AlI of the patients from whom the virus was isolated survived the infection
and were subsequently shown to have acquired specific antibody, but 2 other
cases, diagnosed clinically and pathologically respectively, died.

fter t.
Appendix III
-170 -
mosquito iso1ates were 14 from Haema~o~us spp., 1 from Sabethini, and 1 from
a mixed pool of Aedes and Sabethini.
Four of the mosquito strains carne from
the Oriboca epidemic area, the others from Lazaropolis do Prata (Para State) ,
Macapa (Amapa Territory) and the Utinga forest near Belem.
a gap of 5 years, a further iso1ate was made in 1960 from the
blood of a sentinel monkey stationed 94 km. from Belem along the Belem-
Brasilia highway trace43 In the same year 3 strains of 17D vaccine virus
were iso1ated at the BVL from recent1y inocu1ated persons, and in 1962 there
were 2 further iso1ations from poo1s of Haemagogus spp. caught at km. 87 and
9
94 respective1y, of the Be1em-Brasi1ia road. ln December 1964 a strain was
isolated from the blood of a sentinel monkey in Utinga forest, Belem, more
than 9 years since the last isolation there (from Haema~o~us), and in view
of the continuous surveillance of the are a during that time, yellow fever
seems to have been actually absent, probably brought back by the movements
of monkeys44 Eradication of jungle yellow fever from Amazonia is impossible,
but continued~. aegypti control has taken care of any possibility of urban
epidemics, and vaccination is carried out when indicated in rural locations
Louis encephalitis
This is another virus which causes epidemics in the USA but has not
been isolated from mau in Amazonia. Isolations from Sabethes belizarioi
(BeAr 23379) and from Gi~anto1ae1aps mites combed from an Oryzomys rat have
been made in the region45. There have a1so been iso1ations from a poo1 of
Culex (Culex) mosquitoes (probably ~
) dec1arator) and 3 forest birds
9
and from a Didelphis marsupial and a sentinel mouse group . As with EEE,
up to 17% of forest birds were found to have HI antibody confirmable by
N test, but not rodents or marsupiaIs.
35
35

Appendi:x II!
Ilheus
-171 ~
~s d~I
~~.,
b?
Th1s v1rus has been 1so1ated 1n Amazon1a tw1ce from fever pat1ents
(BeH 7445), twice from the b1ood of sentine1 monkeys, and 19 times from
9 17
mosquitoes, principa11y Psorophora ferox' .No wi1d vertebrate from the
region has as yet yielded virus. RI ant1bod1es have been found 1n 10% of
rodents, 15% of mar8upia1s, 10% of Tamarin monkeys, 4% of BradvDus sloth8,
11% of Co1umb1aa111na b1rds and 4/13 11zards from the Braz111an Amazon13
Qut no N tests have been done
Bussuoua.ra
The prototype (BeAn 4073) was isolated at the BVL from the blood of
a sentinel ~Qqatta beelzebul. monkey, which later died with liver pathology
similar to that produced by yellow fever infection46 Other strains have
come from sentine1 mice, Proechimys rats (which gave strain BeAn 4116, most
commonly used at the BVL), and Culex and Mansonia mosquitoes. Proechimvs
has RI antibody rates rising to a peak of as much as 70% in the middle of
the year29 A single N test positive bird serum has been reported35, and
68% of ColumbiRallina birds and up to 8% of some species of marsupial were
HI positive13, but no N tests were dane on these.
GROUP C
!bis group was first established to contain 5 new virus types isolated
at the BVL17. Later a sixth type was discovered47 which completed a cycle
of serological interrelationships25 Several other types have since been
described from Trinidad and Panama, but only one of these, Nepuyo virus,
~.
JK
0.J)('-
9 --has
also been encountered in Amazonia . Ant1gen1c variat1on within the group
L.R
has been discussed by Shope~-. Three strains of a Caraparu variant (BeH 5546)
76
have a1so been is01ated from man and sentine1 mice at the BVL .
., ,~

itoes, rker, es.
Appendix rn
-172
Tab1e 11 summarizes the data obtained at the BVL on these 7 virus
Caraparu virus emerges as the most commonly isolated type, and in
fact 1t 1s the most frequent1y found of a11 v1ruses 1n the Be1em area.
Together with the closely related Itaqui virus, Caraparu virus has been
isolated more often from rodents than marsupiaIs, has higher antibody rates
in rodents than in marsupials, and is apparently carried almost exclusively
by Culex mosquitoes. Murutucu virus shares all these characteristics
except that its HI antibody rate in marsupials (range 8-23%) is higher than
in rodents (range 1-8%). In contrast, Oriboca virus has been isolated from
a wide range of mosquito species, and the ranges of its HI antibody rates
in rodents and marsupiais coincide, although the highest rates for both
Oriboca and Murutucu antibodies are found in the arboreal genera of marsuâre
pials (Calluromvs and Marmosa). There ~ scantier data for the remaining
and its range of HI antibody rates is higher in marsupiais, although
overlapping with the rodent range. Marituba and Nepuyo a180 appear to infect
marsupiais preferentially, and HI antibody rates to the former are higher
in arboreaI than in forest fIoor marsupiaIs. A mosquito has still to be
found carrying Marituba virus. Marituba virus was isolated from a BVL
but it is possible that this could have been a natural infection
The epidemiological significance of the isolations of Murutucu virus from
a sloth and a pool of ticks collected from marsupiaIs beIonging to several
genera 18 uncerta1n.
Antibody rates for the Group C viruses require careful interpretation
in view of their close serological interrelationships. Thus HI & N tests
for Oriboca antibodies also detect Itaqui antibodies, and vice versa.
9

,
Appendix III
173
Similarly Murutucu HI & N tests will algo detect Marituba antibody, and
Caraparu tests will detect Apeu antibody. However, significant differences
have been found between the rates obtained by HI test for, e.g., Murutucu
and Caraparu antibody on the same batch of sera13 Shope ~ !!.35 report
a rate of 3% for RI antibody to Caraparu virus in forest birds, and a
rate of 6% Murutucu antibody has been found in monkeys13
was dane
but no N testing
Transmission to mice from naturally infected mosquitoes caught in the
Amazon forest has been achieved for Oriboca virus with a Culex (Melanoconion)
sp., probably portesi, and for Nepuyo virus with Culex of unidentifiable
49 50
species ' A.e
The histopathology of sentinel mice naturally infected with Group C
viruses has been described by De Paola ~ ~.51 who found, in addition to
the expected encephalitic lesions, liver lesions in the rnice infected with
Itaqui and Caraparu viruses. Liver lesions have algo been found in howler
monkeys (Alouatta) experimentally infected with the same 2 viruses, and in
infant mice inoculated and naturally infected with Murutucu and Oriboca
52 80
viruses '
GUAMA GROUP
This group was also first established on the basis of new isolates
from the BVL17,S3 Guama and Catu viruses are the second and third most
frequently encountered viruses at the BVL (after the Group C agent Caraparu)
Three serologically related virus types are separable as the BeAn 8582
(Capim) group or subgroup, and a fourth type, BeAn 7722 (Mirim) crossreacts
with Guama antigen but not antiserum by RI test13 Fifty-twa isalates af
the Guama group made at the BVL have 80 far defied further typing.

Appendi:x III
-174 -
Tab1e 12 summarizes the data from the BVL on the 7 types.
Only Guama
and Catu have been isolated from man, and serological data for man is only
avai1able for BeAn 8582 (Capim), where 1/37 people was HI positive at Borba,
on the Rio Madeira, and 327 from other parte of the Brazilian Amazon were
negative13 It appears that a11 the viruses except BeAn 20076 (Bushbush-1)
and BeAn 7722 (Mirim), for which the data are insufficient, are more closely
involved with rodents than with marsupials, and with Culex rather than with
other species of rnosquitoes.
Iso1ations of Guama and BeAr 12590 (Moju)
viruses from pools of male Phlebotomus spp. and of Guama from nestling birds
(Troglodytes) are of doubtful validity13 The high antibody rates in
marsupiaIs for these same 2 types are in the arboreal genera CaIIuromys
and Marmosa.
One strain of BeAr 12590 (Moju) was iso1ated froro an unidenti-
fied farest rat9, ather strains af the same virus fram the viscera af a
Proechimys rat and Mansonia spp. trapped in Amapa Territory, and a strain
of Catu virus froro the salivary glands of a bat (~~~ obscurus) algo
froro AmapalO, alI the other Amazonian strains af these viruses being fraro
Para State.
HI antibodies for BeAn 8582 (Capim) in rodents have been shown
to disappear rapid1y, so that the rate of 13% for Proechimys represents on1y
recent infections; Oryzomys have a much lower antibody rate than Proechimys
for this virus and for BeAn 10615 (Guajara), and no N antibodies were de-
9 13 61
tected to the latter in 88 marsupiaIs tested' , .Oryzomys is more
common1y found with viraemia to BeAr 12590 (Moju) than is Proechimys9,13
Transmission to laboratory mice by the bite of naturally infected
mosquitoes caught in the Amazon forest has been successful with 4 of the
virus types, as follows:

Appendix III
Virus Mosquito trans~ission~ Reference
Guama Culex (M.) tae!1.iopus
Culex B17 1
BeAn 20076 (Bushbush?)
Culex spp.
1
BeAn 8582 (Capim) Culex spp.
21
BeAn 7722 (Mirim) Culex (M.) tae~iopus
The histopathology of laboratory mice natural1y and experimentally infected
TODA & SHOPE 49
TODA 50
.51 52
w1th viruses of the Guama group has been described ' ; the lesions were limited
to the central nervous systern.
Bunyarnwera Group
Four types and a complex of closely related strains belonging to this group
have been isolated in Amazonia.
Guaroa virus has on1y been iso1ated from man in the Amazon va11ey (BeH 12208)
a1though iso1ations from Anophe1es have been made in panama54 and the coastal region
55 -
of Colombia The human isolations were from a liver biopsy specimen and 4 gera
from patients with fever and prostration56-58. An HI antibody survey of the
population of the Amazon va11ey gave an overa11 rate of 18%59
60% among the Mundurucu Indians of the upper Rio Tapajoz.
13
with a maximum af
Birds from forests
near Be1em had an HI antibody rate of 0.3%.13 No N antibody tests have been dane
virus
Type BeAr 7272 (Maguari) was at first identified as a strain of Cache Va11ey
60
17, but is separab1e by HI and N test .It has on1y been iso1ated from
mosqui toes, once each from poo1s of Anophe1es nimbus , ~~e_s_Je~coce1aenus and
9
Ae.scapu1aris , and from a mixed poo1 containing the 1ast-named species but with
17
a majority of Psorophora ferox Human HI antibody rates average 6.5% over the

Appendix liI
Kairi vírus has been isolated
176
from a pool of Ae. scapularis (BeAr
17
8226)
a pool of Sabethini, a pool of Wyeomyia spp., a sentinel monkey, a Saimiri
9
monkey, and an Oecomys rat. Ruman RI antibody rates for the Brazilian Amazon
outside Be1em (which was not tested) average 3%, with a peak of 7% at Eirunepe
13
on the Rio Jurua , but N tests have not been dane.
BeAr 32149 (Sororoca) virus has been iso1ated on1y from mosquitoes (Sabethini)
co11ected on human bait on tree p1atforms at Km. 87 & 94 from Belem on the Belem-
Brasilia highway9. The 6 isolations were alI made in 1962, since when there have
been no more recoveries, and no serum survey data are available, largely because it
has not been possible to produce a haemagglutinating antigen froro these strains
The Wyeomyia complex is represented in the Brasilian Amazon by at least
2 sub-types, distinguishab1e on1y by cross-N testing, alI iso1ated from mosquitoes,
principa11y Sabethini, but a1so from a mixed poo1 of ~edes9,17.
types of these are BeAr 278 (Tucunduba) and BeAr 671 (Taiassui.)
The Belem proto-
From these data it appears that the viruses of the Bunyamwera group active in
Amazonia have cycles involving Aedes & sabethine mosquitoes and arboreal hosts,
probably monkeys.
California Group
The only representative of this group so far found in the Amazon is Melao
vírus (TRVL 9375,) which has been recovered there only from mosquitoes, namely
from a mixed poo1 containing Psorophora ferox (Be1em prototype BeAr 8033,) a poo1
--17 9,
af Aedes scapularis and a pool of P. ferox . Two iso1ations reported from a
17 were found on reexamination to
sentinel cebus monkey and a young saimiri monkey
be strains of Kairi virus
62
the Bunyamwera group .
antigen has precluded serum surveys.
9
-the California group is serologically related to
Lack of success in producing a high titred haemagglutinating

odents, rates;
Appendix !II
-177 -
Phlebotomus Fever Group
Five af the virus types af this graup are faund in Para State
of them having been isolated initially at the BVL, and these 4 have not
been found outside Amazonia. Although serologically related to the
viruses of phlebotomus fever, none has been isolated from Phlebotomus,
63
and only Itaporanga virus has so far been isolated from any arthropod.
Mansonia venezuelensis. Itaporanga vírus has algo been ísolated from
4 groups oi sentinel mice (Belem prototype BeAn 64582) and a Calluromys
marsupial in the Brazilian Amazon, and HI antibody is common in arboreal
genera of marsupiaIs, and in forest birds (up to 10%)35, but absent
from forest floor mammals and open field birds in the Belem area, sug-
gesting that transmission is confined to the forest canopy 13
4
Bats of
at least 4 species belonging to 3 genera have algo been found with HI
antibodies near Belem, but N tests have not been dane
Icoaraci virus (BeAn 24262) has only been isolated from forest
principally Proechimys, which species has RI antibody rates of
up to 56%, whereas other species of rodents and marsupiaIs have Iower
HI antibody has a1so been found in repti1es and s10ths, but not
in man in the Be1em area64 The remaining 3 types are: BeH 22511
(Candiru) from a febrile human, BeAn 46852 (Anhanga) from the organs of
a Cho1oepus s1oth, and BeAn 47693 and 67744 (Bujaru) frorn the b1ood of
9,10,13
Proechirnys rats The on1y ser. frorn the Arnazon va11ey found to
react by HI with BeH 22511 (Candiru) antigen have been from one human
13 35
and one lizard ,and 2 birds, one of which aIs o had N antibody .On
the other hand, HI antibody to BeAn 47693 (Bujaru) virus was found in

Appendix ll!
178
13/53 peop1e at Labrea, on the Rio Purus, and in 139/164 Proechimys
tested, as we11 as in up to 14% of forest rodents and marsupia1s13, but
no N tests have been made.
Simbu Group
Two types of this group have been isolated at the BVL. Oropouche
was obtained from a pool of Aedes serratus and the blood of a sloth,
Bradypus tridacty1us (BeAn 19991,) both co11ected on the Be1em-Brasi1ia
highway, and 15 strains were iso1ated from man in 1961 during an epidemic
in Be1em65, when it is ca1cu1ated that the virus probab1y infected
11,000 persons, but without any known deaths13 One probable laboratory
infection which was severe and required prolonged convalescence occurred
in a female laboratory technician at the BVL9. The second type,
BeAn 84785 (Utinga) was iso1ated in 1965 from another specimen of
Bradypus tridactylus captured at the Utinga forest. Belern; it is clearly
distinguishable serologically from Oropouchet although related13
BeAn 24232 (Piry) Group
Strain BeAn 24232 (Piry) was iso1ated in 1960 from the viscera of
9
a Metachirops opossum trapped in the Utinga forest, Be1em . No other
iso1ations have been made in nature. There is c1inica1 and sero1ogica1
evidence of a case of 1aboratory infection in 1964 at the BVL61.
60
WHITMAN has passaged the vírus seria11y in mosquitoes by inocu1ation.
N antibodies have been found in 3% of marsupiaIs and 3% of rodents
61
tested .
Turlock Group
Turlock vírus was isolated for the first time in South America in
91961,
from a sentine1 mouse group in the forest near Be1em (BeAn 32260) .

Appendix III
-179
It was next isolated from 2 birds of different species captured in the
same farest in 1964, and 5% af farest bird gera tested that year had
HI antibodies, many confirmed by N test, in contrast to only one HI
35
reactor among 211 forest rodents and marsupia1s .
Anopheles A Group
A sing1e strain (BeAr 35112) was iso1ated in 1961 from a poo1 of
Anophe1es nimbus co11ected from the Be1em-Brasi1ia highway9. It appears
to be serological1y identical to TRVL 10076 (Lukuni) virus from Trinidad60,77
c.hanguinola Group
Strain BeAn 28873 (Irituia) was iso1ated in 1961 from an Oryzomys
rat trapped at Km 92 from Belem on the Belem-Brasilia highway, and 4
Some of these could be identical to other strains of the Changuinola
54
group isolated in Panama
Timbo Group
Six strains af Timba vírus (BeAn 41787) and 3 strains af Chaca
vírus (BeAn 42217) have been iso1ated from Ameiva ameíva ~~y~ 1izards
at the BVL, and a fourth strain of the latter was isolated from a
Kentropyx calcaratus lizard66. They are considered to be arthropod-
borne viruses on the basis of their pathogenicity for mice and their
sensitivity to desoxycho1ate, a1so Chaco has been successfu11y passaged
through mosquitoes by inocu1ation
60

Appendix III
-180 -
Ungrouped Arboviruses
The following strains, serologically unrelated to any known
arbovirus, are assumed to be arboviruses because of their sensitivity
to desoxycholate and the fact that they have been successfully pas-
saged serially in mosquitoes by inoculation60: Tacaiuma, BeAn 27639
(Acara,) Marco.
Tacaiuma (BeAn 73) virus was originally isolated from the blood
of a sentinel monkey in the Oriboca forest near Belem17 It has since
been found 3 times in pools of Haema~o~us spp. caught in Para State9
and once from mosquitoes of the garoe genus caught in Amapa Territory
HI antibodies, mostly confirmed by N test, have been found in Para
State in man (0.5%,) horses (24/48) and rodents (0.8%)13, and in
forest birds (6%)35, but not in marsupiaIs, cows, sheep, bats, edentates
13
monkeys or lizards .
Strains of BeAn 27639 (Acara) virus have been isolated from 6
groups of sentinel mice and 2 Nectomys aquaticus amazonicus water rats
9 13
in Para State' , and N antibody has been found in Nectomys (2/18,)
Proechimys (12%,) Oryzomys (4%) and marsupiaIs (3%)61.
Straíns of Marco vírus (BeAn 40290) were isolated 4 times from
Ameiva ~ya ameiva lizards at Belem, but nothing is known oi the
prevalence oi antibodies in lizards or other vertebrates66
The next 2 types are desoxycholate sensitive and were isolated
from mosquitoes. The single known strain of BeAr 40578 (Jurona) virus
carne frorn a poo1 of Haernagogus spp. captured in 1962 at Km 87 frorn
Belem on the Belem-Brasilia highway
9
and abaut 20% af farest birds
have HI antibody3S. This antibody has not, however, been confirmable
10

Appendix III
-181
cross-reacting
by N testing and rnay be due to &eft-~eeeti-;e/Sirnbu or Bunyarnwera group
antibodies13. Two strains of BeAr 50117 (Ternbe) virus carne from
~opheles .nimbus caught at Km 87 and 94 but nothing is known of the
9
occurence of antibodies in vertebrates .
Strain BeAn 58058 was isolated from the blood of an Oryzomvs rat
captured in the Utinga forest, Be1em in 1963 and it appears to be
serologically related to Cotia virus9, but behaves differently in
67
tissue culture Cotia virus , which has been isolated many times
in Sao Paulo State, is assumed to be an arbovirus on the basis of its
recovery from sentinel mice and its sensitivity to desoxycholate.
MISCELLANEOUS VIRUSES
Tacaribe Group
68
This serological group includes the following viruses Tacaribe,
isolated Eram bats and possibly Eram mosquitoes in Trinidad; Amapari,
isolated from forest rodents and their ectoparasites in Brazil;
Machupo, from rodents and human cases of the highly fatal Bolivian
haemorrhagic fever; and Junin, isolated from rodents, their ectoparasites
and human cases af Argentinian haemarrhagic fever. These forro a
geographic series, with a transition somewhere in Brazil between the 2
southern t'ypes which are highly pathogenic for rnan, and the 2
equatorial types which, as far as 1s at present known, do not involve
man
Fourteen isolations of Amapari vírus frorn Oryzornys_goeldii and
Neaco~ guianae, and one frorn Garnasid rnites cornbed frorn infected
Oryzomys, had been made from Amapa Territory, Brazil, up to the end of
196510,.69. AlI but one of the rodent isolations have been from pooled

~ppendix li!
and the vírus can probably remain for at least 26 days ín the
of an apparently healthy Oryzomys, since one rat of this species
held alive for that length of time after capture before being
and its viscera yielded Amapari virus.
viru8 ha8 80 far been isolated only from the Beni region
Bolivia. which is part of the Amazon basin.
isolations from
human cases and 9 from the spleens of the semi-domestic rodent
callosus have been described70,71. fine effort in logistics
involved in airlifting a field laboratory and personnel frorn
to San Joaquin, in the Beni, where the townspeople were sickening
haemorrhagic fever at the rate of 10 a week, with case fatalityrates
up to 20%72. The virus was first isolated in infant hamsters
from autopsy material70, and the writer had the privilege of being
present in the fie1d 1aboratory at San Joaquin at the time in 1963
",hpn t"hi ~ ~tt~~P-~~ was a~hip-vp-d. * Subsequently the rodent Calomys was
and a rodent contrai operation was iaunched in the town in
with US Army assistance; 2 weeks after the operation had coveredthe
ares. human cases of haemorrhagic fever ceased dramatica11y73.
studies on the virus and discussion of its epidemiology have
74been
published .
Ungrouped viruses
dístínct vírus types have been isolated from mammals of the
Amazon forest. which are desoxycholate sensitive but unrelated
serologically to any arbovirus, and there is no evidence as to their mode
nf t-T:;n~mi~~inn. One is BeAn 27326 (Pacui,) isolated 7 times from
*~hil~~ in TPrpint nf a traveI ~rant frorn the Rockefeller Foundation.

III
9
Oryzamys , which species has a N antibady rate af 38%, whilst
Appendix The this Four micet Other liver-bIood kidney. ENTOMOLOGICAL
Up mosquitoes,
Isolates been Mayaro, mites
-183
Proechimys has a rate of 31% and Nectomys and forest marsupiaIs have
lower rates61. It does not multiply in mosquitoes after inoculation60.
other i8 BeAn 67949 from the blood of a bat (Carollia subrufa)13;
produces acidophilic intranuclear and intracytoplasmic inclusions
80in
mouse liver and brain .No antibody data are yet available.
Unidentified agents
agents producing CPE in cebus kidney tissue culture but not
in chick embryo, Aotus or Alouatta monkey kidney cultures or infant
were isolated at the Instituto Evandro Chagas tissue culture
laboratory from the following material:
AN 28512, domestic cat, brain;
AN 28531-6, Nectomys, brain-1iver poo1; AN 28667-9, opossum, brain-
pool; CM 3025, sentinel mouse, bIood.
agents producing
CPE were found in uninoculated cultures of .Tamari~, Alouatta and Cebus
They are non-pathogenic to baby mice. and may belong to thesimian
virus series75
NOTES
to the end of Ju1y 1964, a total of 992,339 arthropods, main1y
had been processed for virus isolation at the BVL, in
20,758 pools9,50, and these had produced 279 strains of virus (excluding
9 EMC strains.)
from arthropods other than mosquitoes have
(through December 1965) one of Murutucu vírus from íxodid ticks,
4 of the Changuinola group and one each of Moju and Guama (doubtful)
from ~lebotomus spp., and 4 different viruses from pools of gamasid
ectoparasitic on rodents:
SLE, Cocal and Amapari.

Appendix III
-184
Several mosquito isolates were made from pools of mixed species, or even
of mixed genera; these are listed in Tables 7-10 under the commonest
species in the pool.
The workers in the Bolivian Beni inoculated over 28,000 arthropods
(most1y Trombicu1idae and Ixodidae, with on1y 125 mosquitoes) into
mice, without iso1ating Machupo virus; a 1arge number of mosquito poo1s
73
remained to be processed as of November 1964 .
The taxonomy of the genus Culex is extremely difficult in the
Belem area. Many females can be separated as species, but until the cor-
responding males can be successfully bred from them, their identification
is in doubt. and therefore these species are referred to by BVL codes as
Culex Bl, B7, etc.
Attempts at rearing eggs from wild-caught females
at the BVL are succeeding, and as a result it appears that the Culex
referred to in early BVL reports as C. (Tinolestes) mo;uensis, and in
later ones as Culex B9, is actually C. (Melanoconion) portesi78. It
is still necessary to determine whether the species identified from
the Belem area as C.(c.) coronator is in fact C.(C.) mollis. References
in BVL reports to C. (Tinolestes) sp. should read Culex spp. (subgenus
undetermined,)50 and those to C.(C.) virgultus should read ~.)
declarator79
SUMMARY
This review covers viruses isolatedt or for which there is
serological evidence of occurrence, in the Amazon basin. These include
the viruses of variola, herpes, influenza, mumps polia, encephalomyo-
carditis, and rabies, several adeno and Coxsackie viruses, mouse polio-
vírus, Newcastle and foot-and-mouth dísease víruses, some 60 dífferent

Appendix III
185
types af arbavirus, and a number af miscellaneaus viruses including
the aetiological agent of Bolivian haemorrhagic fever.
Most of these
agents have been isolated at the Belem Virus Laboratory of the Instituto
Evandro Chagas, Be1em, Para, Brazi1, since its opening in 1954. Over
2,000 strains of arboviruseshave been isolated at that laboratory,
from man
wild and sentinel anirnals, and arthropods. Tables show the
sources of these isolates by species, and maximum antibody rates
encountered in man and wild vertebrates.

1. 3. figs. Hyg.
~:372-96, li:620-625,
Appendix III
ANON., 1965
186 -
REFERENCES
Proposals and recommendations of the provisional committee for
nomenclature of viruses (P.C.N.V.)t Ann. Inst. Pasteurt 1Q2:625-
637
2. ANDREWES c., 1964
Arboviruses, ~ "Viruses of Vertebrates", Williams & Wilkins Co
Ba1timore, 401 pp.
CAUSEY O.R. & MAROJA, 1959
Isolation of yellow fever virus from man and mosquitoes in the
Arnazon region of Brazil, Arner. J. Trop. Med. Hyg. ~:368-371
4. SCHAEFFER M., GAJDUSEK D. C., BROWN A. & EICHENWALD H., 1959
Epidernic jungle fevers among Okinawan colonists in the Bolivian
rain forest. I. Epidemiology, Ibid.
5. GAJDUSEK D. C., ROGERS N. & BANKHEAD A. S., 1959
14 figs.
Serological survey of viral and rickettsial diseases among jungle
inhabitants of the upper Amazon basin, Pediatrics 11:121-131, 6
6. MACKENZIE R. B., BEYE H. K., VALVERDE L. & GARRON H., 1964
Epidemic hemorrhagic fever in Bolivia I. A preliminary report of
the epidemiologic and clinical findings in a new epidemic area in
Sauth America, Amer. J. Trap. Med.
2 figs

8. 14. 15.
Appendix trr
7. CAUSEY C. E., 1958
187 -
Or~anizacao de laboratorios centrais e de campo, Rev. Servo Esp.
Saude Pub.. 1&:35-41
LOBO G. G. B., 1962
Isolamento de poliovirus de fezes de criancas durante um surto de
poliomielite ocorrido na cidade de Belem, no periodo de outubro de
1961 a marco de 1962, Ibid. 12:71-79
9. CAUSEY O. R., personal communication
10. PINHEIRO F. P., personal communication
AZEVEDO M. C., personal communication
12. JONKERS A. H.t SHOPE R. E.t AITKEN T. H. G. & SPENCE L.t 1964
Cocal virus, a new agent in Trinidad related to vesicular stomatitis
vírus, type Indiana, Amer. J. veto Res. li:236-242
SHOPE R. E. & ANDRADE A. H. P., personal communication
CAUSEY O. R., SHOPE R. E. & LAEMMERT H., 1962
Report of an epizootic of encephalomyocarditis virus in Para,
Rev. Servo Esp. Saude Pub. g:47-50
DICK G. W. A., 1960 Êl WOODALL J. P., 1965
Summary of virus isolations from àrthropods at this Institute, to
the end af 1964, E. Afr. Vírus Res. Inst. Rep. 14:16-20
PALACIOS C. A., personal communication

Appendix III
-188
CAUSEY O. R., CAUSEY C. E., MAROJA O. H. & MACEDO D. G., 1961
The isolation of arthropod-borne viruses, including members of
two hitherto undescribed serological groups, in the Amazon region
af Brazil. Amer. J. trapo Med. Hyg. .!Q.:227-249
CAUSEY O. R. & CAUSEY C. E.. 1958
Inquerito sorologico na Amazonia, Rev. Servo Esp. Saude Pub.
10:143-150
CAUSEY O. R. & THEILER M., 1958
Vírus antíbody survey on gera of resídents of the Amazon Valley
in Brazil, Amer. J. Trap. Med. Hyg. I: 36-41
CAUSEY O. R., 1958
Reservatorios e transmissores, Rev. Servo Esp. Saude Pub
~:133-136
CAUSEY O. R., 1958
Procedencia dos arbovirus isolados de 1954 a 1957 em Be1em, Ibid.
10:82-85
22. CAUSEY C. E. & CAUSEY O. R., 1958
Situacao em reIacao as especies conhecidas e ao esquema de CasaIs,
dos arborvirus isolados em Belem, segundo os sintomas observados
nos camundongos inoculados, Ibid. 10:78-80
23. MAROJA O. M. & CAUSEY C. E., 1958
o emprego da fixacao de complemento como teste para situacao dos
arbovirus amazonicos nos grupos sorologicos de CasaIs e Brown, Ibid.
.!Q.:69-71

Appendix III
24. SHOPE R. E., 1962
-189
The serological identification of arthropod-borne viruses. Ibid.
Q:33-38
25. SHOPE R. E. & CAUSEY O. R., 1962
Further studies on the serological relationships of Group C
arthropod-borne viruses and the application of these relationships
to rapid identification of types, Amer. J. Trop. Med. Hyg
.!!.:283-290
26. CAUSEY O. R
The isolation of virus from natural and sentinel hosts in the
Amazon Valley, Rev. Servo Esp. Saude Pub. 12:25-31
27. CAUSEY C. E. & CAUSEY O. R., 1962
The arthropod-borne viruses of Brazil in relation to world groups.
12:9-13
28. CAUSEY C. E.,
The role of small mammals in maintenance of arboviruses in the
Brazilian Amazon forests, An. Microbiol. 11:119-121
29. SHOPE R. E
The use of a micro hemagglutination-inhibition test to follow
antibody response after arthropod-borne virus infection in a com-
munity of forest animaIs, Ibid. 11:167-171,2 figs.
30. SHOPE R. E.
The serological response of animaIs to virus infection in Utinga
forestt Be1emt Brazi1t Atas Simp. Biota Amazonicat Be1emt Parat 1966

31. ~3. 15.
~4 .
~ppendix -190 5ROPE, rhe arthropod-borne
32. ~AUSEY [solamento
venezuelana'l
[solamento :'este" ~AUSEY ~pizootic )ara, ;HOPE rhe ;pecial ~razil i6. ;AUSEY [nvestigation
~iver 5:1017-1023
:ausative
III
-
R. E., CAUSEY O. R., ANDRADE A. R. P. & THEILER M., 1964
Venezuelan equine encephalomyelitis complex of group A
viruses, including Mucambo & Pixuna from the
~azon region of Brazil, Amer. J. Trop. Med. Hyg. !..;!:723-727
O. R. & MACEDO D. N. G., 1958
e identificacao do virus da l'Encefalomielite Equina
no Para, Rev. Servo Esp. Saude Pub. 19-:72-74
~AUSEY O. R., MACEDO D. N. G. & CAUSEY C. E., 1958
e identificacao do virus da 'Encefalomielite Equina
no Para, Ibid. hQ.:75-77
O. R., SHOPE R. E., LAEMMERT H. & SUTMOLLER P., 1962
eastern equine encephalitis in the Braganca region in
Ibid. !,?:.:39-45
R. E., ANDRADE A. H. P., BENSABATH G., CAUSEY O. R. &
dUMPHREY P. 5., (submitted)
epidemiology of EEE, WEE, SLE, and Turlock viruses, with
reference to birds, in a tropical rain forest near Belem,
O. R. & MAROJA O. M., 1957
of an epidemic of acute febrile illness on the
Guama in Para, Brazil, and isolation of Mayaro virus as
agent, Amer. J. Trap. Med. Hyg.

Appendix III
37.
38.
-191
CAUSEY O. R., MAROJA O. & AZEVEDO M. C., 1958
Epidemia pelo virus "Mayaro" no estado do Para,
Rev. Servo Esp. Saude Pub. 10:152-154
PORTERFIELD J. s., 1961
Cross-neutralization studies with Group A arthropod-borne viruses,
World Hlth. Org.
24:735-741
SCHMIDT J. R., GAJDUSEK D. C., SCHAEFFER M. & GORRIE R. H., 1959
Epidemic jungle fever among Okinawan colonists in the Bolivian
rain forest.
Isolation and characterization of Uruma virus,
a newly recognized human pathogen, Amer. J. Trop. Med. Hyg.
487
40. CAUSEY O. R., CASALS J., SHOPE R. E. & UDOMSAKDI S., 1963
42.
Aura & Una, two new group A arthropod-borne viruses, Ibid
777- 781
BENSABATH B. & ANDRADE A. H. P., 1962
Anticorpos para arbovirus no soro de residentes na cidade de
Belem, Para, Rev. Servo Esp. Saude Pub. 12:61-69
SCAFF L. M., 1958
Vacinacao antiamarilica, 19-:155-160
43. LAEMMERT H. & CAUSEI O. R., 1962
A febre amarela na regiao amazonica, Ibid. 12:51-54
8:479

50.
Panamer.
Para, Trap. mosquitoes,
Appendix III
192 -
44. BENSABATH G't SHOPE R. E't ANDRADE A. H. P. & SOUZA A. P. 1966
,li- '..
Recuperacion de virus amarilico, procedente de un mono centin~~, en
45
Ias cercanas de Belem, Brazil, BoI. Oficina Sanit.
P..Q.:187:192
CAUSEY O. R., SHOPE R. E. & THEILER M., 1964
Isolation of St. Louis encephalitis virus from arthropods in
Brazil, Amer. J. Trap. Med. Hyg. g:449
46. GOMES G. & CAUSEY O. R., 1959
Bussuquara, a new arthropod-borne virus, Proc. SOCo Exp. Bio1.
lQ1.:275-279
47 SHOPE R. E., CAUSEY C. E. & CAUSEY O. R., 1961
Itaqui virus, a new member of arthropod-borne Group C, Amer. J
Med. Hyg.
48. SHOPE R. E., 1965
10:264-265
Antigenic variation among arboviruses, Ciencia e Cultura
11.:30-32
49. TODA A. & SHOPE R. E., 1965
Transmission of Guama & Oriboca viruses by naturally infected
Nature (Lond.) ~:304
TODA A.. personal communication

Appendix li!
-193
DE PAOLA D., DUARTE F. & LOBO M. B., 1963
Histopatologia da infeccao natural por arbovirus do grupo C e
Guama, An. Microbiol. ..!l:211-216
DE PAOLA D., 1964
Contribuicao ao estudo da patologia das arboviroses. Tese
apresentada para o concurso de docencia-livre de Anatomia e
Fisiologia Patologicas da Faculdade de Ciencias Medicas da
Universidade de Estado da Guanabara, Rio de Janeiro, 91 pp.,
19 figs.
53. WHITMAN L. & CASALS J., 1961
The Guama group: a new serological group of hitherto undescribed
19..:259-263
Immunological studies, Amer. J. Trop. Med. Hyg.
PERALTA P. H., GALINDO P. & SHELOKOV A., 1961
Virus isolates from Panamanian mosquitoes and sandflies, Fed
20:436
55. SANMARTIN C.
Symp. on Arboviruses of the California Complex & the
Bunyamwera Group. Smo1enice. 1966
56. A., VIANNA C. M. & SILVA E. S.. 1963
Ocorrencia de casos de encefalomielite em doentes oriundos da
Estrada Belem-Brasilia. Rev. Cienc. Biol. 1:27-31

Appendix III
CAUSEY O. R., SHOPE R. E. & RODRIGUES A., 1962
Isolamento do virus Guaroa do figado por biopsia percutanea de
um caso humano com paralisia, Rev. Servo Esp. Saude Pub.
12:55-59
CAUSEY O. R., 1966, ~ WOODALL J. P.
Human infection with arboviruses of the Bunyamwera group, Proc.
Symp. on Arboviruses of the California Complex & the Bunyamwera
Smo1enice, 1966
CAUSEY O. R., 1960, ~ HORSFALL F. L. & TAMM 1.,1965
"Viral and Rickettsial Infections of Man", 4th edn., p. 662
B. Lippincott Co., Phi1ade1phia, 1282 pp
WHITMAN L., personal communication
BENSABATH G., personal communication
62. WHITMAN L. & SHOPE R. E., 1962
The California complex of arthropod-borne viruses and its relation-
ship to the Bunyamwera group through Guaroa virus, Amer. J. Trop.
Hyg.
11:691-696
TRAPP E. E., ANDRADE A. H. P. & SHOPE R. E., 1965
a newly recognized arbovirus from Sao Paulo State,
Proc. Soco Exp. Biol.
64. CAUSEY O. R. & SHOPE R. E.. 1965
a new virus related to Naples Phlebotomus Fever virus,
118:420-421

Appendix 111
65. PINHEIRO F., PINHEIRO M., BENSABATH G., CAUSEY O. R. & SHOPE R. E.,
-195
Epidemia de virus Oropouche em Belem, Rev. Servo Esp. Saude Pub
JJ:.:15-23
CAUSEY O. R., SHOPE R. E. & BENSABATH G. 1966
Timbo & Chaco. newly recognized arboviruses from lizards
of Brazil, Amer. J. Trop. Med. Hyg.
li: 239-243
67. LOPES O. S., LACERDA J. P. c., FONSECA I. E. M., CASTRO D. P.,
FORATTINI O. P. & RABELLO E. S. 1965
Cotia virus: a new agent isolated from sentinel mice in Sao Paulo,
Ibid. 1!i:156-157
SHELOKOV A., 1965
Hemorrhagic fevers in the Americas: a perspective, Ibid.
14:790-792
69. PINHEIRO F. P., SHOPE R. E., ANDRADE A. H. P., BENSABATH G
G. V. & CASALS J., (submitted)
Amapari, a new virus of the Tacaribe Group from rodents and
mites of Amapa Territory, Brazil
70. JOHNSON K. M., WIEBENGA N. H., MACKENZIE R. B., KUNS M. L.,
TAURASO N. M., SHELOKOV A., WEBB P. A., JUSTINES G. & BEYE H. K.,
Virus isolations from human cases of hemorrhagic fever in Bolivia,
Soco Exp. Biol. ill:113-118

Appendix III
-196 -
JOHNSON K. M., KUNS M. L., MACKENZIE R. B., WEBB P. A. &
YUNKER C. E.. 1966
Isolation of Machupo virus from wild rodent Calomys callosus,
J. Trap. Med. Hyg. 103-106
72. MACKENZIE Ro Bo, 1965
Epidemiology of Machupo virus infection. Pattern of human
M. L., 1965
San Joaquin, Bo1ivia, 1962-1964, Ibid. li: 808-813
Epidemiology of Machupo virus infection. 11. Ecological & con-
traI studies of hemorrhagic fever, Ibid. 14:813-816
74. VARIOUS AUTHORS, 1965
Symposium on some aspects of hemorrhagic fevers in the Americas,
L4_: 789-818
HENDERSON J. R., personal communication
76. CASALS J. & WHITMAN L. 1961
Group C, a new serological group oí hitherto undescribed arthropod-
borne viruses. Immunological studies. J. Trop. Med. Hyg.
10:250-258
77. ANDRADE A. H. P.. personal communication
AITKEN T. H. G.&GALINDO P. (in press)
On the identity of Culex (Melanoconion) portes i Senevet & Abonnec
1941 (Diptera, Cu1icidae) Proc. Ent. Soco Wash.

Appendix III
-197 -
79. STONE A., KNIGHT K. L. & STARCKE H., 1959
'IA synoptic catalog of the mosquitoes of the world (Diptera,
Culicidae. ) II Washington (Thomas Say Foundn., VaI. VI.)
80. L. B., personal communication

Appendix III

Appendix III
Table 2
Arbovirus isolations from man and maximum antibody rates,
V i r u s
Mayaro
Mucambo
Ilhéus
Yellow fever
Apeú
Caraparú
Itaquí
Marituba
Murutucú
Oriboca
Catú
Guamá
Guaroa
Oropouche
H22511 (Candirú)
15 types
Be1em Virus Laboratory 1954-65
No. isolates Antibody rate a)
Field Lab. HI NT
a) HI and N rates mar be on different collections of gera
b) includes 3 isolations of l7D from vaccinated persons.
c) these could have been natural infections.
9
6
3
24b)
3
8
1
2
4
4
9
3
5
15
1
Total 97
2
-
-
-
-
-
-
1c)
-
-
-
-
-
1c)
-
4
60%
34%
-
-
15%
15%
-
-
3%
4%
4%
3%
18%
-
1/253
10%
>20%
36%
45%
-
-
-
-
-
-
-
-
-
19%
-

Appendix III
200 -
Table 3
Arbovirus isolations from sentinel animaIs,
Belem Virus Laboratory, 1954-65
Virus Mouse l Monkev I Total
EEE
Mucambo
Bussuquara
I1heus
SLE
Ye11ow fever
Apeu
Caraparu
Itaqui
Marituba
Murutucu
Nepuyo
Oriboca
Catu
Guama
AR 12590 (Moju)
AN 20076 (Bushbusl'0
AN 8582 (Capim)
AN 10615 (Guajara)
AN 7722 (Mirim)
Kairi
Turlock
Itaporanga
AN 27639 (Acara)
Tacaiuma
isolated
34
37b)
9
1
-
11
292
137
11
25
3
42
147
181
89
8
18
33
8
3
2
4
1095
22
a)- algo 2 isolates from sentinel chickens
b)- ane af these was a white rat
7
30
lc)
1
2
15
31
4
13
18
20
8
22
1
1
1
41a)
67
10 112
26
323
141
24
43
3
62
155
203
89
8
18
33 9
1
3
2
4
1
175
16 26

~t~al
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Appendi.1: III
-201 -
Table 4
Virus isolates from rodents and maximum antibody rates
Be1em Virus Laboratory 1954-65
Virus No. I Antibody Ik$olates~
No.
EEE
Muc amb o
Pixuna
Bussuquara
Caraparu
Itaqui
Murutucu
Nepuyo
Oriboca
Catu
Guama
AR12590 (Moju)
AN8582 (Capim)
ANIO615 (Guajara)
Icoaraci
AN47693 (Bujaru)
AN28873 (Irituia)
AN27326 (Pacui)
AN27639 (Acara)
AN58058 (Cotia?)
EMC
Amapari
-
1
1
1
2
2
3
2
2
rate I
1%HI*
24%HI
24%HI
43%HI
4/12HI
14%HI
2/12HI
23%HI
39%HI
30%HI
34%HI
3%HI
8%HI
16%HI
2/28NT
1/6 NT
-
1
4
1
15
5
2
3
1
1
414
7
7
1
9
2
4
I~~o~a~es
Antibody
rat~_~
4%HI
43%HI
70%HI
29%HI
26%HI
35%HI
3%HI
28%HI
45%HI
43%HI
43%HI
13%HI
10%NT
28%HI
85%HI
31%NT
14%NT
3/34NT
No. IAntibod:)f
isolatesl rate j;t,solates
1
13
2
2
1
1
6
12
8
1
7
1
7

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(AN70100) 1 1(ANl1497)
1
Appendix III
-203-
Table 6
Virus isolations from other wild vertebrates
Be1em Virus Laboratory 1954-65
Virus Vertebrate sp.
No.
isolates Type
Mvrmotherula hauxwelli
Mucambo (AN88995) Pipra ervthrocephala
(AN69763)
bird
bird
4~tomolus infuscatus 1
bird
(AN59768) Myrrnotherula hauxwelli
(AN70092)
Thamnomanes caesius
Murutucu (ANl1249) Bradypus tridactylus 1
sloth
Catu (AN94103) Molossus obscurus 1 bat
Kairi
bird
bird
Saimiri sp. 1 monkey
Oropouche (AN19991) Bradypus tridactylus 1 sloth
(Utinga) (AN84785)
Bradypus tridactylus
Turlock (AN72648) Myrmotherula hauxwelli 1 bird
(Anhanga
(AN73173) Thamnophilus amazonicus 1 bird
sloth
(AN46852) Choloepus brasiliensis
1 sloth
Timbo (AN41787) Ameiva ameiva ameiva 6 lizard
Chaco (AN42217) p.meiva ameiva ameiva-
3 lizard
Kentropyx calcaratus
1 lizard
Marco (AN40290) Ameiva ameiva ameiva 4
lizard
(unnamed) (AN67949) Carollia subrufa 1 bat
EMC (AN18124 J acana .1 acana
1 bird
(AN18360) Monasa ni~rifrons 1 bird
(AN18436) Guira ~uira
(AN18368) Urubutinga sp.
bird
bird

Appendi:x:
III
V i r u s
Aedes spp.
-204
Table 7
Arboviruses isolated from ~ mosquitoes
A. arborealis
Be1em Virus Laboratory, 1954-65
A. argyrothorax
A. falvus
A. leucocelaenus
A. scapularis
*one pool also contained one specimen each of Ae. scapularis, P. ferox and an
unidentified sabethine.
A. septemstriatus
A. serratus
A. sexlineatus
A. taeniorhynchus
Total
Total spp.
Aura 5 5 1
EEE 3 3 1
Mucambo 2* 2 1
Una 1 1 2 2
Ilheus 1 1 1 1 3 7 5
Apeu 1 1 2 2
Oriboca 1 1 2 2
Guama 1 1 2 2
AN7722 (Mirim) 1 1 1
Kairi 1 1 1
A7272
(Maguari)
1 1 2 2
Melao 1 1 1
Wyeomyia 1 1 1 3 3
Oropouche 1 1 1
Total 2 2 1 1 4 4 1 15 1 3 34
No. types
isol.
2 2 1 1 4 4 1 8 1 1 14

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declarator
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spp.
V i r u s
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EEE 2a) 1 3a) 1 1a) 12a) 20a) 6
Mayaro 1 1 1
Mucambo 3 1 8 1 1 14 5
Bussuquara 6 1 1 2 1 11 5
SLE 1 1 1
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Itaqui 3 5 1 1 1 1 12 6
Murutucu 1 1 1 1 4 4
Nepuyo 1 1 1
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Catu 20 1 21 2
Guama 4 1 1 19b) 1 1 1b) 1 2 31 9
AR 12590 (Moju) 1 2 1 1 5 4
AN 8582 (Capim) 18 8 26 2
AN 10615 (Guajara) 2 1 1 4 3
AN 20076 (BushBush?) 2 2 1
AN 7722 (Mirim) 1 1 2 2
Itaporanga 1 1 2 2
Total 49 12 17 57 2 5 2 9 4 5 16 178
No. Types isolated 15 5 7 9 2 5 2 8 4 5 4 20
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M.
M. Psorophora P. P.
Virus
Total Total spp.
spp. spp. arribalzagai titillans venezuelensis albipes ferox lutzi
Mayaro 25 25 1
Mucambo 1 1 1 3 3
Una 1 9 10 2
Bussuquara 1 1 2 2
Ilheus 2 9 11 2
Yellow fever 16 16 1
Oriboca 1a) 1 1 1 4 4
-206 -
Catu 1 1 1
Guama 1 3 1b) 5 3
AR 12590 (Moju) 1 1 2 2
Melao 2 2 1
Itaporanga 1 1 1
AR 40578 (Jurona) 1 1 1
Tacaiuma 4 4 1
Wyeomyia 1 1 1
Maguari 1c) 1 1
Total 47 3 2 2 9 3 22 1 89
No. types isol. 5 3 2 2 7 2 5 1 16
a) pool contained one Psorophora sp. b) identified only as Group Guama c) mixed pool also containing Aedes and
Mansonia

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Tab1e Isolates Sentinel 141 14 13%c)
3% 15% Rodents 12% 5% 35% 3% MarsupiaIs 8% 22% 17% 23% 11%
Appendix !Ir
from:
-208-
11
Group C virus isolations and antibody rates,
Be1em Virus Laboratory. 1954-65
Oriboca
AN 17
Itaqui
AN 12751
Caraparu
AN 3994
Apeu I
AN 848,
Maritube
AN 15
Man 4 1 3 3
animaIs 62
Murutucu
AN 974 :
Nepuvo
a)
321 26 24 43 3
Rodents 2 5 8 6
MarsupiaIs
Aedes 2 2
Culex 6 12
Other mosquitoes 6 1
HI antibodies (max.) in
Man
a) Be1em prototype BeAn 10709
Total 83 160 351 33 27 59b) 4
28% 26% 43%
23%
b) also 2 isolations from a sloth (Bradypus) and a pool of Ixodid ticks
HI rates must be interpreted with caution in the absence of N test data
1
4% 4%
27%

Appendix III
Isolates from:
-209 -
Tab1e 12
Guama group virus isolations and antibody rates,
Guama I
BeAn 277
Be1em Virus Laboratory, 1954-65
Catu
BeH 151
Man 3 9
BeAr 1259°
(Moju)
1
HeAn 85821
(Capim)
,BeAn 10615'I
(Guajara)
BeAn 20076 JjeAn 7722
:Bushbush?) (Mirim)
Sentinei animais 203 155 89 18 33 8 7
29 12 18 7
5
Aedes 2 1
Culex 31 21 5 26 4 2 2
Other mosquitoes 5 2
Total 278 198 114 52 38 10 10
HI antibodies (max.) in:i
Man