Characterization of Serum Complement Activity in Three Species of Crocodilians from Southeast Mexico.

To characterize the serum complement innate immune system in three species of crocodilians native to southeast Mexico. Methodology: Plasma collected from three wild crocodilian species native to southeast Mexico were exposed to sheep red blood cells (SRBCs) to measure hemolysis, which is used as an indication of serum complement immune activity. Results: Incubation of different volumes of plasma from Crocodylus acutus, Crocodylus moreletii, and Caiman crocodilus resulted in a volume-dependent increase in SRBC hemolysis. However, while maximum hemolysis for C. acutus and C. moreletii were both approximately five-fold higher than that of Ca. crocodilus. A kinetic study revealed that the hemolysis was rapid, with near-maximum activity recorded at 30 min for C. acutus and C. moreletii. However, Ca. crocodilus activity exhibited a significant increase (P<.5) only between one and two hours. A thermal analysis showed that the SRBC hemolysis was maximal at temperatures to which these species thermoregulate. The thermal profiles were similar for all three species, although the activity was lower for Ca. crocodilus (P<.01). The SRBC hemolysis was strongly inhibited by mild heat treatment (56°C,30 min) and also by EDTA, indicating that the hemolytic activity was probably due to the presence of crocodilian serum complement activity. The EDTA-inhibited activity was restored by the addition, of a 20mM excess of Ca2+or Mg2+, but not Fe2+orCu2+, thus exhibiting the specific need for Ca2+orMg2+. Conclusions: The serum complement activities of C. acutus and C. moreletii are much higher (P<.01) than Ca. crocodilus, which may be related to the maximum sizes and increased aggressive natures of C. acutus and C. moreletii.

Humoral immune alterations caused by lead: studies on an adult toad model

There is evidence that environmentalmetal levels affect the immune function. In the particular case of the impact of heavy metals, information available suggests that the immune system is a target for low-dose Pb exposure. Among vertebrates it was shown that amphibians are capable of forming antibodies against a variety of antigens, causing several responses such as anaphylactic response and rejectinggrafts. In this study, the production of antibodies was assessed against sheep red blood cells (SRBC) in the anuran Bufo arenarum after six weekly injections of sublethal doses of lead (50 mg.kg-1, as lead acetate). Natural antibodies (natural heteroagglutinins)were also quantified against SRBC. Both assessments were carried out employing an ELISA method developedto this end, measuring absorbance (A). For natural anti-SRBC antibodies in both control (C) and Pb treated (T) toads, there was a non significant tendency to increase the initial absorbances (C initial: 0.69+0.39 A; T initial: 0.54+0.30 A), relative to those registered at the end of the experiments (C final: 0.89+0.49 A; T final: 0.76+0.31A); the T/C ratios also did not show changes. The only significant difference was found between initial and final samples from lead-treated toads (p<0.014). The immune anti-SRBC antibody levels of toads immunized with SRBC showed a significant lower increase (p<0.05) in lead-treated animals (T final: 0.66+0.36 A), as compared to control toads (C final: 0.91+0.50 A) at the end of the experiment. It was thus concluded thatthe changes due to the assayed doses of Pb in the levels of antibodies cannot be explained on the basis of only one singleaction mechanism of the metal, but as the result of a conjunction of effects over different immunocompetent cell subpopulations. These different responses suggest that factors affecting animals exposed to a foreign stimulus are different from those influencing the response of wild animals.

Existe evidencia de que los niveles de metal ambientales afectan la función inmune. En el caso particular del impacto de metales pesados, la información disponible sugiere que el sistema inmune es un blanco para la exposición a bajas dosis de Pb. Entre los vertebrados, se ha mostrado que los anfibios son capaces de formar anticuerpos contra una variedad de antígenos, que causan diversasrespuestas, tales como respuesta anafiláctica y rechazo de injertos. En este estudio, la producción de anticuerpos fue evaluada contra eritrocitos de oveja (EO) en el anuro Bufo arenarum, luego de seis inyecciones semanales de dosis subletales de plomo (50 mg.kg-1, como acetato de Pb). Los anticuerpos naturales (heteroaglutininas naturales) fueron también cuantificados contra EO. Ambas evaluaciones fueron llevadas a cabo empleando un método de ELISA desarrollado a este fin, midiendo laabsorbancia (A). Para los anticuerpos anti-EO naturales, tanto en sapos controles (C) como en sapos tratados con Pb (T), hubo una tendencia significativa a incrementar las absorbancias iniciales (C inicial: 0,69+0,39 A; T inicial: 0,54+0,30 A); la relación T/C tampoco mostró cambios. La única diferencia significativa se encontró entre las muestras inicial y final de los sapos tratados con plomo (p<0,014). Los niveles de anticuerpos anti-EO inmune de sapos inmunizados con EO mostraron un bajo incrementosignificativo (p<0,05) en los animales tratados con plomo (T final: 0,66+0,36 A), al compararse con sapos control (C final: 0,91+0,50 A) al final del experimento. De este modo, se concluye que los cambios debidos a las dosis analizadas de Pb en los niveles de anticuerpos no pueden explicarse sólo sobre la base de un único mecanismo de acción del metal, sino como resultadode una conjunción de efectos sobre diferentes subpoblaciones de células inmunocompetentes. Estas diferentes respuestas sugieren que los factores que afectan los animales expuestos a un estimulo externo son diferentes...