Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias
Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias
Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias
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16 t h International Congress on Animal <strong>Reproduction</strong><br />
Poster Abstracts 65<br />
P112<br />
Genomic characterization of Arcanobacterium pyogenes<br />
isolates recovered throughout the puerperium of dairy<br />
cows<br />
Silva, E 1 *; Gaivão, M 1 ; Leitão, S 1 ; Jost, B 2 ; Carneiro, C 3 ; Vilela, C 3 ; Costa, L 1 ;<br />
Mateus, L 1<br />
1Department of <strong>Reproduction</strong> and Obstetrics, C.I.I.S.A., Faculty of Veter<strong>in</strong>ary<br />
Medic<strong>in</strong>e, Portugal; 2 Department of Veter<strong>in</strong>ary Science and Microbiology,<br />
University of Arizona, USA; 3 Department of Microbiology and Immunology,<br />
C.I.I.S.A., Faculty of Veter<strong>in</strong>ary Medic<strong>in</strong>e, Portugal<br />
In dairy cows, the establishment and persistence of the puerperal<br />
uter<strong>in</strong>e <strong>in</strong>fection is related to the characteristics of the bacteria that<br />
colonize the uterus. The cl<strong>in</strong>ical signs and reproductive consequences<br />
of the uter<strong>in</strong>e disease are generally attributed to the presence of a few<br />
pathogens, ma<strong>in</strong>ly Arcanobacterium pyogenes (A. pyogenes) alone or<br />
<strong>in</strong> synergy with other bacteria. In or<strong>de</strong>r to i<strong>de</strong>ntify factors that might<br />
be associated with the establishment and persistence of uter<strong>in</strong>e<br />
<strong>in</strong>fection, we characterized A. pyogenes isolates obta<strong>in</strong>ed throughout<br />
the puerperal period of dairy cows from two unrelated herds.<br />
The isolates were recovered from 26 cows (8 had normal puerperium<br />
and 18 <strong>de</strong>veloped uter<strong>in</strong>e <strong>in</strong>fection). The genomic characterization of<br />
A. pyogenes (n= 57) was performed by BOX-PCR typ<strong>in</strong>g and by<br />
screen<strong>in</strong>g of putative virulence factors through conventional PCR.<br />
Consi<strong>de</strong>r<strong>in</strong>g a genomic similarity of at least 86%, a total of ten<br />
different clonal types (5 <strong>in</strong> each herd) were i<strong>de</strong>ntified. These clonal<br />
types were totally herd-specific and a s<strong>in</strong>gle type appeared to<br />
predom<strong>in</strong>ate <strong>in</strong> each herd. Only one clonal type was not associated<br />
with endometritis, while four others were always associated with<br />
uter<strong>in</strong>e disease. However, 5 clonal types were present <strong>in</strong> cases of both<br />
normal puerperium and endometritis. Throughout the puerperal<br />
period, 1 to 3 clonal types were sequentially or simultaneously<br />
isolated from the same animal.<br />
In or<strong>de</strong>r to i<strong>de</strong>ntify virulence factors associated with the <strong>de</strong>velopment<br />
of <strong>in</strong>fection, all isolates were tested for the presence of 8 putative A.<br />
pyogenes virulence genes: plo (encod<strong>in</strong>g pyolys<strong>in</strong>), nanH and nanP<br />
(encod<strong>in</strong>g neuram<strong>in</strong>idases), cbpA (encod<strong>in</strong>g a collagen-b<strong>in</strong>d<strong>in</strong>g<br />
adhes<strong>in</strong>) and four different fimbrial genes (fimA, fimC, fimE, fimG).<br />
The 8 genes were present <strong>in</strong> 66% of the isolates, 5 of 8 genes (plo,<br />
nanH, nanP, cbpA, fimA) were <strong>de</strong>tected <strong>in</strong> all isolates, fimE <strong>in</strong> 97%,<br />
while fimC and fimG were only present <strong>in</strong> a subset of isolates. There<br />
was no association between BOX-PCR type, presence of the above<br />
genes and <strong>de</strong>velopment of uter<strong>in</strong>e <strong>in</strong>fection.<br />
The results of this study suggest that the type of A. pyogenes may not<br />
be a <strong>de</strong>term<strong>in</strong>ant factor <strong>in</strong> the <strong>de</strong>velopment of uter<strong>in</strong>e <strong>in</strong>fection. Host<br />
<strong>in</strong>tr<strong>in</strong>sic factors and/or the synergism between A. pyogenes and other<br />
bacteria may play a more relevant role <strong>in</strong> the establishment of<br />
puerperal <strong>in</strong>fections.<br />
This research was fun<strong>de</strong>d by the grant POCTI/CVT/48773/2002 from<br />
“Fundação para a Ciência e Tecnologia”. Maria Elisabete Silva is a<br />
post-doc fellow from FCT.<br />
P113<br />
Bacteriological and histological studies of cow’s uterus<br />
without and with reta<strong>in</strong>ed fetal membranes<br />
Skuja, S 1 *; Antāne, V 1 ; Feldmane, L 2<br />
1Faculty of Veter<strong>in</strong>ary Medic<strong>in</strong>e, Latvian University of Agriculture, Latvia;<br />
2Department of Pathological Anatomy, Riga Strad<strong>in</strong>s University, Latvia<br />
Introduction Retention of fetal membranes is economically important<br />
disturbances dur<strong>in</strong>g the postpartum period <strong>in</strong> cattle <strong>in</strong> Latvia. Several<br />
methods have been suggested for the treatment of reta<strong>in</strong>ed fetal<br />
membranes, and controversial results have been published. The aim of<br />
the study was to <strong>in</strong>vestigate the postpartum period of cows without<br />
and with reta<strong>in</strong>ed fetal membranes.<br />
Materials and methods Bacteriological samples of the cow’s uterus<br />
and biopsies of the uterus mucous membranes were collected from 29<br />
Holste<strong>in</strong> cows. The bacteriological exam<strong>in</strong>ation was performed <strong>in</strong><br />
cows with and without reta<strong>in</strong>ed fetal membranes (RFM) two, 14, 40 –<br />
50 days postpartum (PP). On day 40-50 all cows were biopsied from<br />
the uterus endometrium and the uterus status was estimated<br />
histologically. Accord<strong>in</strong>g to the 3rd stage of parturition process<br />
(expulsion of fetal membranes), cows were divi<strong>de</strong>d <strong>in</strong>to the follow<strong>in</strong>g<br />
groups:Control group – fetal membranes were expelled <strong>in</strong> 6-12 h<br />
PP;Group 1 – cows with RFM, removed manually and treated;Group<br />
2 – cows with RFM, it was not removed manually.Group 2 cows with<br />
health disturbances (fever, milk fever, mastitis) were treated<br />
a<strong>de</strong>quately.<br />
Results Bacterial isolates of uterus on day two PP <strong>in</strong> all cows (100%)<br />
conta<strong>in</strong>ed a wi<strong>de</strong> spectrum of microorganism associations: <strong>in</strong> 41% of<br />
cases there were s<strong>in</strong>gle isolates – E. coli 33%, Streptococcus spp. 5%<br />
and Clostridia spp. 3% , and <strong>in</strong> 59% there were mixed isolates – E. c.<br />
27%, Streptococcus spp 3%, Clostridia spp 26%, Staphylococcus spp.<br />
3%. On day 14 PP, 93% of cows had microorganisms <strong>in</strong> the uterus. In<br />
addition to the above f<strong>in</strong>d<strong>in</strong>gs there was one isolate of A. pyogenes <strong>in</strong><br />
the cow with RFM that was removed manually two days PP. In the<br />
next bacteriological exam<strong>in</strong>ation (40-50 days PP) no bacteria were<br />
found, and <strong>in</strong> 84 days PP after two times of artificial <strong>in</strong>sem<strong>in</strong>ation the<br />
cow became pregnant. In group 2 cows, which were treated, the<br />
number of isolated bacterial species <strong>in</strong>creased on day 14 PP. At the<br />
same time cows of group 2, which were untreated, the number of<br />
isolated bacterial species rema<strong>in</strong>ed the previous one, and the number<br />
of isolates <strong>de</strong>creased. On day 40-50 PP, bacterial isolates were found<br />
<strong>in</strong> 86.2% of cows. Cows with a normal parturition process were 50%<br />
with s<strong>in</strong>gle bacterial isolates (Streptococcus spp. and<br />
Corynebacteriom spp.). There were no differences between the<br />
bacterial species and their number of group 2 treated and untreated<br />
cows. In 82.8 % of all <strong>in</strong>vestigated cows, histological f<strong>in</strong>d<strong>in</strong>gs showed<br />
evi<strong>de</strong>nce of mild to mo<strong>de</strong>rate endometritis.At this time, 33.3% ( 8<br />
cows) become pregnant.<br />
P114<br />
I<strong>de</strong>ntify<strong>in</strong>g AI bulls associated with poor fertility and high<br />
rates of embryo loss<br />
Sun<strong>de</strong>, J. 1 * and Refsdal, AO. 2<br />
1Team Sem<strong>in</strong> BA, P.O. Box 8146 Dep., N-0033 Oslo, Norway; 2 Geno<br />
Breed<strong>in</strong>g and AI association, N-2326 Hamar, Norway<br />
The relationship between fertility and semen quality is not always<br />
clear. Occasional AI bulls may show poor fertility even though semen<br />
passes quality test<strong>in</strong>g. These bulls represent a potential loss for the<br />
dairy farmer, and may even possess un<strong>de</strong>sirable genetic traits that<br />
could affect fertility <strong>in</strong> future generations. It is therefore important<br />
that these bulls are i<strong>de</strong>ntified at an early stage <strong>in</strong> the breed<strong>in</strong>g<br />
programme as soon as <strong>in</strong>sem<strong>in</strong>ation data are available for analysis.<br />
Norwegian Red (NRF) is the dom<strong>in</strong>ant dairy breed <strong>in</strong> Norway and<br />
semen is distributed by Geno, a farmer-owned cooperative. Close to<br />
100% of <strong>in</strong>sem<strong>in</strong>ations are reported back to Geno and 95 % of<br />
farmers also report data on calv<strong>in</strong>gs, stillbirths and culls through the<br />
Norwegian Dairy Herd Record<strong>in</strong>g System. These data allow<br />
calculation of accurate non-return rates (NRR) and calv<strong>in</strong>g rates (CR)<br />
on sire basis. At present, bulls with NRR60 < 65 % after approx. 1000<br />
first <strong>in</strong>sem<strong>in</strong>ations of semen for progeny test<strong>in</strong>g are rout<strong>in</strong>ely culled.<br />
However, this parameter alone may not be sufficient to i<strong>de</strong>ntify<br />
animals with potential negative impacts on fertility traits. It was<br />
therefore <strong>in</strong>vestigated whether further analysis of sire fertility data<br />
would yield additional <strong>in</strong>formation.<br />
The data set consisted of 479330 first <strong>in</strong>sem<strong>in</strong>ations on Norwegian<br />
Red (NRF) heifers and cows, with semen from 692 qualified NRF test<br />
and elite sires. Insem<strong>in</strong>ations were performed between January 2000<br />
and Oct 2007. The first day of return-to-service was registered as a<br />
second <strong>in</strong>sem<strong>in</strong>ation with<strong>in</strong> 92 days after first <strong>in</strong>sem<strong>in</strong>ation (pfi), and<br />
a m<strong>in</strong>imum of 1000 first <strong>in</strong>sem<strong>in</strong>ations per bull was required to<br />
qualify for the study.<br />
NRR curves were mo<strong>de</strong>lled us<strong>in</strong>g survival analysis, with day of return<br />
as outcome parameter, adjusted for confound<strong>in</strong>g factors. Sires were<br />
classified as hav<strong>in</strong>g either‘poor’ or ‘good’ fertility if their NRR92<br />
<strong>de</strong>viated from the mean by more than 3 standard <strong>de</strong>viations.<br />
Compared to high fertility sires, poor fertility sires showed a more<br />
abrupt fall <strong>in</strong> NRR from 21 days onwards, suggest<strong>in</strong>g a lower rate of<br />
fertilisation and/or higher <strong>in</strong>ci<strong>de</strong>nce of early embryo <strong>de</strong>ath (before<br />
maternal recognition). These sires also displayed a higher number of<br />
returns-to-service outsi<strong>de</strong> of the normal oestral cycle length. This