Chemical communication in tilapia: a comparison of Oreochromis mossambicus with O. niloticus.

In allopatric speciation species differentiation generally results from different selective pressures in different environments, and identifying the traits responsible helps to understand the isolation mechanism(s) involved. Male Mozambique tilapia (Oreochromis mossambicus) use urine to signal dominance; furthermore, 5ß-pregnane-3α,17,20ß-triol-3α-glucuronide (and its α-epimer, 5ß-pregnane-3α,17,20α-triol-3α-glucuronide), in their urine is a potent pheromone, the concentration of which is correlated with social status. The Nile tilapia (Oreochromisniloticus) is a close relative; species divergence probably resulted from geographical separation around 6 million years ago. This raises the question of whether the two species use similar urinary chemical cues during reproduction. The olfactory potency of urine, and crude extracts, from either species was assessed by the electro-olfactogram and the presence of the steroid glucuronides in urine from the Nile tilapia by liquid-chromatography/mass-spectrometry. Both species showed similar olfactory sensitivity to urine and respective extracts from either species, and similar sensitivity to the steroid glucuronides. 5ß-Pregnan-3α,17α,20ß-triol-3α-glucuronide was present at high concentrations (approaching 0.5mM) in urine from Nile tilapia, with 5ß-pregnan-3α,17α,20α-triol-3α-glucuronide present at lower concentrations, similar to the Mozambique tilapia. Both species also had similar olfactory sensitivity to estradiol-3-glucuronide, a putative urinary cue from females. Together, these results support the idea that reproductive chemical cues have not been subjected to differing selective pressure. Whether these chemical cues have the same physiological and behavioural roles in O. niloticus as O. mossambicus remains to be investigated.

Tilapia male urinary pheromone stimulates female reproductive axis.

Mozambique tilapia males congregate in leks where they establish dominance hierarchies and attract females to spawn in sandy pits. Dominant males store more urine than subordinates and the pattern of urination and the high sensitivity of females to male urine suggest chemical signalling via the urine. Here we show that pre-ovulated and post-spawn females when exposed to dominant male urine increased significantly, in less than 1h, the release rate of the maturation-inducing steroid 17,20ß-dihydroxypregn-4-en-3-one which is maintained elevated for at least 6h. This indicates a pheromonal role for male urine in the synchronisation of spawning. Furthermore, we show that the lack of affinity of 17,20ßP to sex steroid binding globulin explains, at least partly, its rapid release and lack of detection in the blood. Thus tilapia urine involvement in several communication processes confirms that cichlids have evolved a sophisticated chemical signalling system together with their complex visual, acoustic and behavioural displays.

Male urine signals social rank in the Mozambique tilapia (Oreochromis mossambicus).

BACKGROUND: The urine of freshwater fish species investigated so far acts as a vehicle for reproductive pheromones affecting the behaviour and physiology of the opposite sex. However, the role of urinary pheromones in intra-sexual competition has received less attention. This is particularly relevant in lek-breeding species, such as the Mozambique tilapia (Oreochromis mossambicus), where males establish dominance hierarchies and there is the possibility for chemical communication in the modulation of aggression among males. To investigate whether males use urine during aggressive interactions, we measured urination frequency of dye-injected males during paired interactions between size-matched males. Furthermore, we assessed urinary volume stored in the bladder of males in a stable social hierarchy and the olfactory potency of their urine by recording of the electro-olfactogram. RESULTS: Males released urine in pulses of short duration (about one second) and markedly increased urination frequency during aggressive behaviour, but did not release urine whilst submissive. In the stable hierarchy, subordinate males stored less urine than males of higher social rank; the olfactory potency of the urine was positively correlated with the rank of the male donor. CONCLUSION: Dominant males store urine and use it as a vehicle for odorants actively released during aggressive disputes. The olfactory potency of the urine is positively correlated with the social status of the male. We suggest that males actively advertise their dominant status through urinary odorants which may act as a 'dominance' pheromone to modulate aggression in rivals, thereby contributing to social stability within the lek.

Plasma and urine levels of C18, C19 and C21 steroids in an asynchronous fish, the tilapia Oreochromis mossambicus (Teleostei, Cichlidae).

Female and male tilapia, Oreochromis mossambicus, were treated with luteinizing hormone-releasing hormone analogue (LH-RHa) and pimozide (PIM) or with human chorionic gonadotropin (hCG) to stimulate gonadal development and sexual maturation. Plasma (both sexes) and urine (males) samples were collected periodically for steroid analysis by radioimmunoassay. Plasma levels of estradiol-17 beta (3-6 ng/ml) and testosterone, higher in female (up to 25 ng/ml) than in male (6-13 ng/ml; P < 0.05), were in the range of those established in other tilapia species. Plasma levels of the established teleost oocyte maturation-inducing steroids (MIS), that is 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) and 17 alpha,20 beta, 21-trihydroxy-4-pregnen-3-one (17,20 beta,21-P) were low (1-9 ng/ml) and were not different between treated and control fishes at 8, 12, 16, 24, 48, 72 and 96 hr after injection. Furthermore, in male O. mossambicus, 17,20 beta,21-P was undetectable. Plasma levels of 3 alpha,17 alpha,21-trihydroxy-5 beta-pregnan-20-one (3,17,21-P-5 beta) were very high in both sexes (up to 700 ng/ ml), mostly in hormone-treated groups, whose levels were higher than controls (P < 0.05). Urine levels of conjugated 17,20 beta,21-P (glucuronides and sulphates) were not detectable, but those of 17, 20 beta-P (up to 25 ng/ ml) and 3,17,21-P-5 beta (up to 1 microgram/ml) were higher than free 17,20 beta-P and 3,17,21-P-5 beta measured in the plasma of the same animals (P < 0.05). Both LH-RHa + PIM and hCG induced sexual maturation of O. mossambicus (histological data); nevertheless, during that period all measured steroids, either in plasma or urine, almost did not fluctuate. Thus, this study does not make any comment about the MIS of tilapia. Nevertheless, the high levels of conjugated 3,17,21-P-5 beta and 17,20 beta-P in urine suggest a probable pheromone role for these steroids in this species.