Androgens and immune function in male alternative reproductive morphotypes of the peacock blenny Salaria pavo

Species with alternative reproductive tactics offer the opportunity to analyse how behavioural and morphological traits are tuned to produce successfully competing phenotypes within one sex. In the teleost fish Salaria pavo, alternative reproductive tactics are sequential. The older ornamented males compete for access to females by guarding a cavity to which they attract females to spawn. Ornamented males that are found without a nest are called ‘floaters’. Younger mature males which are too small to compete with nest-holders attempt to ‘sneak’ as female mimics into successful nests and release their sperm to fertilize freshly spawned eggs. The alternative tactics in S. pavo are associated with different levels of the androgens testosterone and 11-ketotestosterone, which have been found to suppress immune function in several teleost fish. A field study was carried out to analyse the relationship between these reproductive tactics, androgens and blood levels of lymphocytes as a monitoring method of immune function. We expected highest investment in the immune system in sneakers because these have the lowest androgen levels and functionally because investing in self-maintenance increases their future prospect to switch tactic and to reproduce as a nest-holder. Indeed, the relative count of lymphocytes correlated negatively with body size and thus was highest in sneakers and lowest in nest-holders. In concordance, 11-ketotestosterone and testosterone levels were found to be higher in floaters and nest-holders than in sneakers. However, no correlation was found between individual levels of testosterone or 11-ketotestosterone and lymphocytes. Thus, a trade-off between reproductive traits associated with high androgen levels and immunocompetence might exist at the level of alternative tactics but this might not be explained by acute immunosuppressive effects of circulating androgens

Aggressive behaviour and energy metabolism in a cichlid fish, Oreochromis mossambicus

We have investigated the effect of mirror-elicited agonistic behaviour on oxygen consumption in the Mozambique tilapia, Oreochromis mossambicus (Cichlidae). Males exposed to their mirror image showed higher frequencies of both lateral display and tail-beating and escalated aggression more frequently than males exposed to a transparent glass that was used as a control for the presence of a novel object in the tank. This aggressive response was correlated with an increase in oxygen consumption. Overt aggression was highly correlated with display behaviour and with locomotor activity. Bivariate analyses showed high correlation (explaining about 64% of variation) between overt aggression, locomotor activity and metabolic rates. Weakly positive bivariate correlations between displays and metabolic rates turned spurious after partialling out aggression. The data suggest that energetic costs only emerge late during the conflict, when animals escalate their aggressive behaviour

Intra-sexual variation in male reproduction in teleost fish: A comparative approach

The occurrence of intra-sexual variation in reproduction is a widespread phenomenon in teleosts. One such form of variation consists in the occurrence of alternative male types: males that invest resources in mate attraction and males that exploit the investment of the former males, by trying to sneak fertilizations during spawning. These alternative reproductive tactics can be classified according to their plasticity during the life span of the individuals (i.e., fixed vs. sequential vs. reversible). Furthermore, the differences between morphs within a given species may involve a set of different traits, including reproductive behavior, the differentiation of male morphological traits, and the patterns of gonad tissue allocation and the differentiation of gonadal accessory glands. In this paper, we review the available data on four species exhibiting different types of intra-sexual plasticity in reproduction that have been studied in our lab. The data on the proximate mechanisms, androgens and forebrain arginine-vasotocin (AVT), underlying these alternative tactics suggest that between-morph differences in androgen levels, especially in 11-ketotestosterone, are especially present in species where the alternative male types have evolved morphological traits that are tactic-specific (i.e., sexual polymorphisms) and that differences in AVT appear to be related to between-morph differences in the expression of courtship behavior. Therefore, this comparative approach leads us to propose that the different endocrine systems are involved in the differentiation of different sets of traits that make up alternative phenotypes, and that the differentiation of alternative tactics is not controlled by a single endocrine system (e.g., androgens)

Adjustment of brood size and androgen levels in a teleost species with exclusive male parental care

In maximizing reproductive success, individuals face a trade-off between parental care for their current offspring and investing in androgen-dependent sexual traits to produce further offspring. It has thus been proposed that parental effort would suppress androgen levels. Here, we studied parental effort by manipulating brood size in the peacock blenny, Salaria pavo, a littoral fish species in which males show long periods of paternal care.We focused on the effects of brood size on female spawning rate (measured as number of eggs received in the nest by focal males) and androgen levels. We found a positive linear relationship between brood size and the number of eggs received subsequently. Accordingly, spawning rate increased for males with experimentally enlarged broods while it decreased for males in which brood size was reduced. However, over a longer time interval, brood sizes of both treatments returned to the nonmanipulated brood size, suggesting an effect of additional factors such as male quality. Brood size did not show the expected negative relationship with levels of testosterone and 11-ketotestosterone. On the contrary, testosterone levels were positively correlated with brood size. However, in agreement with the prediction, changes in the level of parental care by manipulation of brood size showed an inverse relationship with testosterone levels. As with brood size, given time, testosterone levels tended to return to nonmanipulated levels. Such changes suggest that males adjust their brood size to an individual value through androgen modulation of courtship or other traits influencing female spawning rate

The arginine-vasotocin and serotonergic systems affect interspecific social behaviour of client fish in marine cleaning mutualism

Many species engage in mutualistic relationships with other species. The physiological mechanisms that affect the course of such social interactions are little understood. In the cleaning mutualism, cleaner fish Labroides dimidiatus do not always act cooperatively by eating ectoparasites, but sometimes cheat by taking bites of mucus from so-called "client" reef fish. The physiological mechanisms in these interspecific interactions, however, are little studied. Here, we focussed on three neuromodulator systems known to play important roles in intraspecific social behaviour of vertebrates to examine their role in clients' interspecific behaviour. We subjected the client fish Scolopsis bilineatus to ectoparasites and the exogenous manipulation of the vasotocin (AVT), isotocin (IT) and serotonin systems to test how this affects client willingness to seek cleaning and client aggression towards cleaners. We found that a single dose of AVT agonist and a selective antagonist caused clients to seek proximity to cleaners, independently of ectoparasite infection. In contrast, in a direct encounter task, the selective blocker of serotonin 5HTreceptors, Ketanserin (KET), made client reef fish more aggressive towards cleaners in the absence of cleaners' bites of mucus. IT did not yield any significant effects. Our results suggest that the AVT system plays a role in social affiliation towards an interspecific partner, while the serotonin system affects clients' acceptance of level of proximity to cleaner fish during interactions. These two systems, therefore, were apparently co-opted from intraspecific social interactions to affect the course of interspecific ones also

Metabolic costs of aggressive behaviour in the siamese fighting fish, Betta splendens

Aggressive conflicts between males are often resolved by means of multiple ritualized agonistic displays without damaging escalation. Apparently, in such cases by using those displays opponents exchange important motivational and physical information on which they base a decision to stay or leave the interaction. In the Siamese fighting fish, the time spent spreading the dorsal fin and erecting the gill coverts predicts who will be the winner or loser of the interaction. Two experiments were carried out to study whether display performance might be costly. First, oxygen consumption was measured during mirror-image stimulation. This experiment showed that oxygen consumption was positively correlated with gill cover erection and dorsal fin spread. In the second experiment, a fight between two opponents was simulated and the oxygen consumption of the expected winner and loser was compared. Metabolic rates were not different between winners and losers before and during the fight, but winners showed higher oxygen consumption in the night after the fight. These results are in accordance with costs of display performance and with longlasting physiological consequences of winning or losing a fight

Regulation of immunocompetence by different androgen metabolites in a blenny with alternative reproductive tactics

In Parablennius parvicornis, small reproductive males with relatively low expression of secondary sexual characters (M morphotype) parasite on the parental investment of the larger nest-holder males which have fully developed secondary sexual characters (M1 morphotype). In comparison with M1 males, M males have relatively low levels of androgens while having high blood cell percentages of lymphocytes and antigen responsiveness. Here we test the hypothesis that androgens are a causal factor for these differences in immunocompetence between morphotypes. After drawing an initial blood sample, males received a silastic implant containing either oil only (C), or oil with testosterone (T) or 11-ketotestosterone (KT). Males were re-caught 2 weeks later for drawing of the final blood sample. KT but not T induced the development of secondary sexual characters in M males. M males treated with KT showed lower swimming activity than the males treated with T or C implants, suggesting that KT also mediates behavioral changes in M males. As expected, blood cell percentages of lymphocytes, but not of granulocytes, were higher in M males than in M1 males. Overall, lymphocyte percentages increased in the C group which might have been a response to the surgery/treatment. In concordance with the hypothesis, lymphocyte percentages were suppressed in males treated with T in comparison with controls. However, no significant change was found in KT-treated males. This suggests that androgens modulate central, morphological and immunological traits by partly independent androgen mechanisms in P. parvicornis

Mounting an immune response correlates with decreased androgen levels in male peafowl, Pavo cristatus

Studies testing the ‘‘immunocompetence handicap hypothesis’’ have focussed on the immunosuppressive effects of androgens. Several recent studies have reported that mounting a humoral immune response might also result in a decrease in circulating androgen levels via a ‘‘negative feedback’’ on the hypothalamus–pituitary– gonadal axis (HPG). The aim of this correlative study was to analyse these immunosuppressive and HPG-suppressive interactions in reproductively active males of the peafowl. We collected blood samples of free living birds before and after challenging the immune system with a non-pathogenic antigen (sheep erythrocytes), and analysed immune parameters and plasma levels of the two main androgens in birds, testosterone and dihydrotestosterone. Males displaying larger versions of the main secondary sexual trait, the long and conspicuously ornamented train, tended to have higher androgen levels and significantly lower circulating levels of leukocytes, indicating that exaggerated ornaments might signal properties of the endocrine and immune system. Actual circulating levels of androgens did not correlate with the plasma levels of leukocytes and the antibody response to SRBC. However, changes in plasma levels of both androgens showed negative correlation with both leukocytes (P\0.1) and SRBC responses (P\0.05). The data therefore support the prediction that activity of the immune system is HPG-suppressive. Such suppression has been proposed to be especially costly during the reproductive season, during which androgens facilitate the expression of exaggerated traits that play an important role in sexual competition

Endocrine correlates of intra-specific variation in the mating system of the St. Peter’s fish (Sarotherodon galilaeus)

The Challenge Hypothesis postulates that androgen levels are a function of the social environment in which the individual is living. Thus, it is predicted that in polygynous males that engage in social interactions, androgen levels should be higher than in monogamous animals that engage in parental care. In this study, we tested this hypothesis at the intra-specific level using a teleost species, Sarotherodon galilaeus, which exhibits a wide variation in its mating system. Experimental groups of individually marked fish were formed in large ponds with different operational sex-ratios (OSR) to study the effects of partner availability on blood plasma levels of sex steroids [11-ketotestosterone (11-KT), testosterone (T), and 17,20B-dihydroxy-4-pregnen-3-one (17,20B-P)] and gonadosomatic index (GSI). Polygyny mostly occurred in the female biased OSR groups. 17,20B-P and gonadosomatic index did not differ among OSR groups. However, 11-KT was high in male biased OSR and positively correlated with aggressive challenges, thereby supporting the central postulate of the Challenge Hypothesis. The results of T were the inverse of those of 11-KT, probably because 11-KT is metabolized from T. 11-KT levels of polygynous males did not differ neither from those of monogamous males, nor from those of males that participated in parental care. These results do not support the expected relationships between polygyny, parental care, and androgen levels. The differences from expectations for 11-KT may be related to the fact that in S. galilaeus, the mating and the parenting phase are not clearly separated and thus, males may still fight and court while they are brooding