Papel de las actividades superóxido dismutasa y catalasa en la ...
Papel de las actividades superóxido dismutasa y catalasa en la ...
Papel de las actividades superóxido dismutasa y catalasa en la ...
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Journal of Fish Diseases 2006, 29, 355–364<br />
PDíaz-Rosales et al. Superoxi<strong>de</strong> dismutase and cata<strong><strong>la</strong>s</strong>e in P. damse<strong>la</strong>e ssp. piscicida<br />
(a)<br />
(b)<br />
1 2 3 4 5 6 1 2 3 4 5 6<br />
Figure 2 Detection of superoxi<strong>de</strong> dismutase<br />
(a) and cata<strong><strong>la</strong>s</strong>e activity (b) in extracts of<br />
Photobacterium damse<strong>la</strong>e subsp. piscicida<br />
(strain EPOY-8803-II) grown un<strong>de</strong>r differ<strong>en</strong>t<br />
conditions. Lane 1: growth until expon<strong>en</strong>tial<br />
phase; 2: stationary growth phase;<br />
3: exposure to hydrog<strong>en</strong> peroxi<strong>de</strong> (20 lm<br />
H 2 O 2 mid-expon<strong>en</strong>tial phase and 2 mm<br />
H 2 O 2 early stationary phase); 4: addition of<br />
methyl violog<strong>en</strong> (0.2 mm) to the culture<br />
medium; 5: addition of 2,2¢-dipyridyl<br />
(100 lm) to the culture medium; 6: addition<br />
of FeCl 3 Æ6H 2 O (100 lm) to the culture<br />
medium.<br />
U SOD/mg prot.<br />
20<br />
15<br />
10<br />
5<br />
0<br />
EPOY-8803-II<br />
Lgh41/01<br />
Figure 3 Superoxi<strong>de</strong> dismutase activity (U mg )1 protein) of<br />
Photobacterium damse<strong>la</strong>e subsp. piscicida strains grown un<strong>de</strong>r<br />
differ<strong>en</strong>t culture conditions. ( ) Growth until stationary phase;<br />
( ) exposure to hydrog<strong>en</strong> peroxi<strong>de</strong> (20 lm mid-expon<strong>en</strong>tial<br />
phase and 2 mm early stationary phase); ( ) culture supplem<strong>en</strong>ted<br />
with FeCl 3 Æ6H 2 O 100 lm and (h) culture supplem<strong>en</strong>ted<br />
with the iron che<strong>la</strong>nt 2,2¢-dipyridyl 100 lm. Data repres<strong>en</strong>t the<br />
mean (SD) of three in<strong>de</strong>p<strong>en</strong><strong>de</strong>nt <strong>de</strong>terminations.<br />
U cat./mg prot.<br />
140000<br />
120 000<br />
100000<br />
80000<br />
60000<br />
40000<br />
20 000<br />
0<br />
EPOY<br />
Lgh41/01<br />
Figure 4 Cata<strong><strong>la</strong>s</strong>e activity (U mg )1 protein) of Photobacterium<br />
damse<strong>la</strong>e subsp. piscicida strains grown un<strong>de</strong>r differ<strong>en</strong>t culture<br />
conditions. ( ) Growth until stationary phase; ( ) exposure to<br />
hydrog<strong>en</strong> peroxi<strong>de</strong> (20 lm mid-expon<strong>en</strong>tial phase and 2 mm<br />
early stationary phase); ( ) culture supplem<strong>en</strong>ted with FeCl 3 Æ<br />
6H 2 O 100 lm and (h) culture supplem<strong>en</strong>ted with the iron<br />
che<strong>la</strong>nt 2,2¢-dipyridyl 100 lm. Data repres<strong>en</strong>t the mean (SD)<br />
of three in<strong>de</strong>p<strong>en</strong><strong>de</strong>nt <strong>de</strong>terminations.<br />
In or<strong>de</strong>r to <strong>de</strong>termine the influ<strong>en</strong>ce of the levels<br />
of SOD and cata<strong><strong>la</strong>s</strong>e activity on the resistance to the<br />
bactericidal activity of sole phagocytes, killing assays<br />
were carried out with a virul<strong>en</strong>t and non-virul<strong>en</strong>t<br />
strain of P. damse<strong>la</strong>e ssp. piscicida. The perc<strong>en</strong>tages<br />
of surviving bacteria after 5 h contact with sole<br />
phagocytes are shown in Fig. 5. It can be observed<br />
that survival of the virul<strong>en</strong>t strain in contact with<br />
phagocytes was significantly higher (P < 0.05) in<br />
all cases compared with the non-virul<strong>en</strong>t strain.<br />
Despite this differ<strong>en</strong>t survival rate, both strains<br />
showed a simi<strong>la</strong>r behaviour <strong>de</strong>p<strong>en</strong>ding on the<br />
bacterial culture condition with highest rates<br />
corresponding in both cases to growth in ironreplete<br />
broths and lowest to growth un<strong>de</strong>r ironlimiting<br />
conditions. In addition, a significant<br />
increase in the survival perc<strong>en</strong>tages was observed<br />
%Survival<br />
100<br />
90<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
EPOY-8803-II<br />
Lgh41/01<br />
Figure 5 Survival perc<strong>en</strong>tage of Photobacterium damse<strong>la</strong>e subsp.<br />
piscicida after 5 h in contact with sole phagocytes. ( ) Growth<br />
until stationary phase; ( ) exposure to hydrog<strong>en</strong> peroxi<strong>de</strong> (20 lm<br />
mid-expon<strong>en</strong>tial phase and 2 mm early stationary phase); ( )<br />
culture supplem<strong>en</strong>ted with FeCl 3 Æ6H 2 O 100 lm and (h) culture<br />
supplem<strong>en</strong>ted with 2,2¢-dipyridyl 100 lm. Data repres<strong>en</strong>t the<br />
mean (SD) of nine wells containing phagocytes from three fish<br />
specim<strong>en</strong>s.<br />
Ó 2006<br />
B<strong>la</strong>ckwell Publishing Ltd<br />
360