Immunoendocrinology and Ecoimmunology in Brazilian Anurans.

This paper reviews several aspects of immunoendocrinology and ecoimmunology in Brazilian species of anurans under investigation for more than a decade, including (1) patterns of annual covariation of circulating steroids, calling behavior and innate immunity, (2) endocrine and immune correlates of calling performance, (3) behavioral and physiological correlates of parasite load, (4) thermal sensitivity of immune function, and (5) endocrine and immunomodulation by experimental exposure to acute and chronic stressors, as well as to endocrine manipulations and simulated infections. Integrated results have shown an immunoprotective role of increased steroid plasma levels during reproductive activity in calling males. Moreover, a higher helminth parasite load is associated with changes in several behavioral and physiological traits under field conditions. We also found anuran innate immunity is generally characterized by eurythermy, with maximal performance observed in temperatures close to normal and fever thermal preferendum. Moreover, the aerobic scope of innate immune response is decreased at fever thermal preferendum. Experimental exposure to stressors results in increased corticosterone plasma levels and immune redistribution, with an impact on immune function depending on the duration of the stress exposure. Interestingly, the fate of immunomodulation by chronic stressors also depends in part on individual body condition. Acute treatment with corticosterone generally enhances immune function, while prolonged exposure results in immunosuppression. Still, the results of hormonal treatment are complex and depend on the dose, duration of treatment, and the immune variable considered. Finally, simulated infection results in complex modulation of the expression of cytokines, increased immune function, activation of the Hypothalamus-Pituitary-Interrenal axis, and decreased activity of the Hypothalamus-Pituitary-Gonadal axis, as well as reduced melatonin plasma levels, suggesting that anurans have a functional Immune-Pineal axis, homologous to that previously described for mammals. These integrated and complementary approaches have contributed to a better understanding of physiological mechanisms and processes, as well as ecological and evolutionary implications of anuran immunoendocrinology.

Thermal sensitivity of innate immune response in three species of Rhinella toads.

Immune activity is temperature-dependent and strongly related to thermal biology in ectotherms. Eurythermic, vs stenothermic, species commonly show a broader range of immune responses. Furthermore, the development of behavioral fever in ectotherms is correlated with improved immune function. Although amphibians generally show restricted capacity for thermoregulation in the field, behavioral fever has been documented in the laboratory for several anurans. However, the match between behavioral fever and improved immune response at fever thermal preferendum has still to be determined in these animals. In this study, we investigate the thermal sensitivity of the innate immune response, as measured by the plasma bacterial killing ability (BKA) against Aeromonas hydrophila, in three species of toads from genus Rhinella (R. schneideri, R. icterica and R. ornata) during their breeding season. Moreover, we show lipopolysaccharide-induced behavioral fever for R. ornata. The three species of toads showed an inverted U-shaped pattern of thermal sensitivity regarding BKA, with a high efficiency of immune response at temperatures around their thermal preferendum. The results partially corroborate the hypothesis that immune function is maximized at fever thermal preferendum, given that two of the species showed a maximal BKA performance temperature closer to fever than their normal thermal preferendum. Toads also showed an eurythermic pattern of immune response (large temperature breadth of BKA performance ≥95%; B95) during the breeding season. This large B95 encompasses much of the ecologically relevant temperatures, with the exception of those exhibited by two species that maintain activity during winter. Lastly, BKA was commonly suppressed at 37 °C, highlighting the importance of choosing ecologically relevant temperatures when conducting in vitro immunological tests.

Behavioral fever decreases metabolic response to lipopolysaccharide in yellow Cururu toads (Rhinella icterica).

Ectothermic vertebrates develop behavioral fever in response to bacterial products, with potential corresponding metabolic costs associated with immune stimulation. Although behavioral fever has been described in several taxa under laboratory conditions, some important questions regarding metabolic response to bacterial products at different temperatures and effectiveness of behavioral fever remain open. Many ectotherms, such as nocturnal anurans, may be active in the field at environmental conditions that restrict thermoregulation during the immune response. How does the metabolic response to bacterial products under ecologically relevant but unfavorable thermal field conditions compare to that measured in fever thermal preferendum? Additionally, are there differences in the partitioning of metabolic costs associated with immune stimulation and Arrhenius effect (biochemical reactions rate) at normal versus fever thermal preferendum? We compared the energy expenditure untreated and LPS-treated yellow Cururu toads (Rhinella icterica) at temperature corresponding to field activity during winter nights, and at normal and fever thermal preferendum. It was hypothesized that the metabolic response to LPS would be proportionally lower at higher body temperatures. To test these hypotheses, we measured temperature in the field during night using agar models, as well as normal and fever thermal preferendum of the toads within a thermal gradient. Subsequently, we measured the toad's metabolic rates at mean agar models temperature, as well as at normal and fever thermal preferendum. Lastly, we calculated the Metabolic response to LPS as the ratio between MRLPS/MRSaline in each of these mean temperatures. Our results show that metabolic rates do not increase in response to LPS at the agar models temperature typical of the winter nights under which theses toads maintain reproductive activity. Moreover, LPS treatment increased the metabolic costs relative to Arrhenius effects at normal thermal preferendum but not at fever thermal preferendum. In this way, metabolic response to LPS was comparative lower at fever than normal thermal preferendum in yellow Cururu toads.

Behavioral, physiological and morphological correlates of parasite intensity in the wild Cururu toad (Rhinella icterica).

Large numbers of parasites are found in various organs of anuran amphibians, with parasite intensities thought to modulate the host's Darwinian fitness traits. Interaction between the anuran hosts and their multiple parasites should modulate the host's phenotypic characteristic, such as those associated with high energetic demand (such as calling effort and locomotor performance), energy balance (standard metabolic rate), and morphological plasticity (as indicated by organ masses). The present study investigated the impact of parasite intensities on the behavioral, physiological, and morphological traits of wild adult male Rhinella icterica (Anura: Bufonidae). We tested as to whether individuals with higher parasite intensities would present: 1) lower vocal calling effort in the field, as well as poorer locomotor performance and body-condition index; and 2) higher standard metabolic rates and internal organ masses. Measurements included: calling effort in the field; standard metabolic rate; locomotor performance; parasite intensity; internal organ masses (heart, liver, kidneys, intestines, stomach, lungs, hind limb muscle, and spleen); and the body-condition index. Results showed a negative association of parasite intensities with locomotor performance, and standard metabolic rate of R. icterica. A positive association between parasite intensities and relative organ masses (heart, intestines and kidneys) was also evident. Toads with higher pulmonary and intestinal parasites intensities also showed higher total parasite intensities.