Kin cohesiveness and possible inbreeding in the mouthbrooding tilapia Sarotherodon melanotheron (Pisces Cichlidae)

Four microsatellite markers were used to study genetic variation among individuals of the mouthbrooding tilapia Sarotherodon melanotheron (Rüppel 1852) caught in separate but adjacent shoals. A comparison was also made with fish from six other localities. Populations originating from riverine environments appear to be panmictic, while samples from open waters such as lagoons showed highly significant heterozygote deficiencies. For instance, at the 33-allele locus SMEL4, 32 homozygous individuals were observed among the 82 individuals from the same lagoon location instead of only five homozygotes expected if random mating occurred. A further assessment of the genetic similarity of individuals within each shoal, validated by robust permutation techniques requiring no precise knowledge of gene frequencies, showed that related individuals tend to aggregate, and suggested that mating occurs preferentially within small groups of kin. Cichlids are often presented as a group of fish where microallopatric speciation takes place. The possible link between kin aggregation, inbreeding and shoaling behaviour we propose here may have important consequences for our understanding of the mechanisms involved in this fast speciation process.     

Functional microsatellite and possible selective sweep in natural populations of the black-chinned tilapia Sarotherodon melanotheron (Teleostei, Cichlidae)

The Prolactin I (PrlI) gene is a key locus involved in fish osmotic regulation. Two microsatellites, PrlI GT and PrlI AC, are present in the promoter region of this gene in tilapias. One of these microsatellite is associated to PrlI gene expression and growth rate. Aquaria reared individuals homozygous for long microsatellite alleles at the PrlI AC locus expressed less PrlI in fresh water than fishes with other genotypes. To assess the occurrence of selection acting on these microsatellites we carried out a survey of their variability compared with four others microsatellites in natural populations. Samples of two tilapia species were studied, Oreochromis niloticus (6 samples) a typically freshwater species inhabiting osmotically stable environments and Sarotherodon melanotheron (5 samples) a brackish water species obliged and able to adapt very rapidly to any salinity changes. For both species the different populations studied exhibited a high amount of differentiation (all F(st) value were statistically significant) at every loci. Only one locus, PrlI AC, was monomorphic for the same allele in all S. melanotheron populations. We have hypothesized that this outlier locus with its unique allele could have experienced a selective sweep that took place in the early stages of the species origin. S. melanotheron represents an example of adaptation (to fluctuant salinity environment) acting indirectly on gene product through its promoter.     

Mouthbrooding in the black-chinned tilapia, Sarotherodon melanotheron (Pisces: Cichlidae): the presence of eggs reduces androgen and estradiol levels during paternal and maternal parental behavior

The black-chinned tilapia (Sarotherodon melanotheron) is considered a paternal mouthbrooder in which the male churns the eggs in his mouth for 14-18 days after they are fertilized. We previously showed that plasma androgen and estradiol concentrations increased significantly between the beginning and end of mouthbrooding. Here we provide evidence that androgen and estradiol are relatively high in prespawning male tilapia, decrease significantly with the initiation of mouthbrooding, and return to and slightly surpass prespawning levels by the end of mouthbrooding. We then test the hypothesis that the initiation of mouthbrooding causes a decrease in androgen and estradiol. We raised single pairs of tilapia in seawater aquaria and conducted two experiments. In the first, we removed eggs from males on the morning of Day 1 of mouthbrooding, followed changes in plasma androgen and estradiol in the afternoon of Day 1 and on Days 6, 11, and 16, and compared them to levels in control males who kept their eggs. Removing eggs from mouthbrooding males on the morning of Day 1 eliminated the decrease in plasma androgen and estradiol observed in the afternoon of Day 1 of mouthbrooding. This demonstrates that the eggs are necessary for the decrease. We observed that, infrequently, the female of the pair would pick up the eggs to brood them in her mouth. In a second experiment, in the afternoon of Day 1, we sampled both the male and female of pairs in which the female brooded eggs. In female-brooding pairs, the females had significantly lower androgen and estradiol levels than prespawning females. The black-chinned tilapia in captivity exhibits both paternal and maternal parental behavior. In both sexes the initiation of mouthbrooding triggers a decrease in androgen and estradiol concentrations. We conclude that the presence of eggs inhibits the pituitary-gonadal axis in both males and females and hypothesize that a chemical signal from the eggs is delaying the initiation of the next brood.     

Osmoregulatory strategies in natural populations of the black-chinned tilapia Sarotherodon melanotheron exposed to extreme salinities in West African estuaries

The effect of salinity was studied in natural populations of the black-chinned tilapia (Sarotherodon melanotheron) from West Africa. This euryhaline species colonizes nearly all coastal environments from bays to lagoons characterized by salinities ranging from fresh water to hypersaline water over 100 ‰. Individuals were sampled during the dry season at several locations characterized by different levels of salinity (3-102 ‰). Their osmotic status and their gills were analyzed. The branchial mitochondria-rich cells (MRC), localized at the basis of the filaments and along the lamellae in fish taken from the saline stations, showed a wide plasticity with significant differences in their number and size. The most striking results were a significant larger area (≈3x) and a higher number (≈55x) of MRC at high salinity (102 ‰) compared to low salinity (3 ‰). The major ion transporters and channels were localized by immunocytochemistry and different expression patterns have been recorded between stations. Despite an increased Na?/K?-ATPase (NKA) α-subunit expression and NKA activity, pointing to an increased monovalent ion excretion, a severe osmotic imbalance was recorded in animals living in hypersaline environments.     

Growth hormone and Prolactin-1 gene transcription in natural populations of the black-chinned tilapia Sarotherodon melanotheron acclimatised to different salinities

The effects of salinity on the expression of genes coding for growth hormone (GH) and prolactin-1 (PRL1) were studied in various natural populations of the black-chinned tilapia Sarotherodon melanotheron from West Africa. Individuals were sampled in June 2005 in six locations in Senegal and the Gambia, at various salinities between 0 and 101. The poorest condition factors were recorded in the most saline sampling site and the best growth in the fish from a marine environment. The pituitary GH mRNA levels were significantly higher in fish adapted to seawater, whereas the PRL1 mRNA levels were highest in fish adapted to fresh- and brackish water. These results show that the PRL1 mRNA levels seem to reflect relatively well the differences in environmental salinity, in contrast to those of GH, which would tend instead to reflect the individual growth in each environment. However, no relation could be found between growth in the hypersaline areas and the expression profile of GH. Although the fish analysed were morphologically identical, the expression of genes coding for GH and PRL1 showed large differences between individuals. This inter-individual variation in gene expression remains poorly understood.     

Effects of salinity on strontium:calcium ratios in the otoliths of the West African black-chinned tilapia Sarotherodon melanotheron in a hypersaline estuary

The biology and the behaviour of fish populations in hypersaline environments are poorly known. The habitat occupation strategy of the tilapia Sarotherodon melanotheron has been studied along a salinity gradient in the Saloum hypersaline estuary in Senegal (salinity between 32 and 100). The individual migratory behaviour has been analysed from otolith strontium-to-calcium concentration ratios from fish sampled in five locations during the 2003 wet season and the 2004 dry season. In the upper part of the estuary (salinities > 50), the Sr:Ca ratio in the otolith showed high variations, from 2.51 to 33.30 × 10⁻³. These maximum observed values have never been reported in the literature. The individual mean of Sr:Ca ratios increased according to the salinity gradient in the estuary, with significantly higher values in the upper part (Sr:Ca mean = 16 × 10⁻³) than in the lower part (salinity < 50, Sr:Ca mean = 12 × 10⁻³). No significant difference in the Sr:Ca mean was observed between locations with comparable salinities. Sr concentration in the water at the different locations was positively correlated with ambient salinity. The mean of Sr:Ca in the otoliths was then in relationship with the level of Sr in the water. Thus, the Sr:Ca ratios in the otolith of S. melanotheron allowed to discriminate the populations of this species and to hypothesize that they did not undertake large scale movements within the estuary.     

Naphthalene-induced changes in carbohydrate metabolism in Sarotherodon mossambicus Peters (Pisces: Cichlidae)

Sarotherodon mossambicus Peters was exposed to naphthalene concentrations ranging from 7.9 to 0.92 mg/l for periods ranging from 4 days to 10 weeks and changes in glycogen, glucose, pyruvic acid and lactic acid contents of liver and muscle were studied. While glucose levels remained unaffected, major effects could be seen in the other constituents following both lethal and sublethal naphthalene treatment. Whereas glycogen and pyruvic acid levels showed considerable reduction, lactic acid levels increased markedly. These changes are indicative of a metabolic stress leading to a shift to anaerobic metabolism. The effects do not appear to be related to the feeding and growth rates of the naphthalene-intoxicated fish.     

Karyotypic changes associated to the dispersive potential on Pomacentridae (Pisces, Perciformes)

When comparing karyotypic diversification between freshwater and marine fish, a notable increase in modifications associated to the former environment can be noticed in relation to the latter one. This fact is related to the great number of physical partitions (barriers) that exist in continental waters. Furthermore, in the marine environment, the barriers to the gene flow are complex and not always identifiable, especially when considering the participation of dynamic factors such as currents and winds associated to the dispersive potentialities of a living group. Within Perciformes fish, the Pomacentridae family stands out from the others for its intimate relation with the coral reefs, as well as for its diversity of forms and biological characteristics, providing an adequate model for studying the influence of the pelagic period of the larval stage upon the karyotypic evolution. In the present work, cytogenetic analyses were carried out on three Pomacentridae species, typical representatives of the coral reefs, Abudefduf saxatilis (2n=48, FN=52, 2 m+2sm+44a), Microspathodon chrysurus (2n=48, FN=64, 6 m+10st+32a) and Amphiprion frenatus (2n=48, FN=92, 14 m+22sm+8st+4a) that have different pelagic larval periods. The analyses between subfamilies showed a trend that the lower the dispersive potential, the greater the quantity of fixed pericentric inversions based on the basal karyotype of the Perciformes.     

Relationship of chin spot size to dominance in the black-chinned mouthbrooding cichlid fish (Sarotherodon melanotheron)

Aggressive interactions of male Sarotherodon melanotheron were studied to determine the communicative value of chin spot size in relation to dominance. In addition, the effects of residency and size on the aggressiveness of this fish were determined. Two-factor analysis of variance was used for frequencies of each modal action pattern for residency and size. Results show that residency played a major role in the outcome of an aggressive interaction, whereas size had little effect. Dominance of each experimental fish was calculated using Barlow & Ballin's (1976) dominance index. A chin spot ratio was obtained by dividing the chin spot area by the total surface area of the individual fish. Simple linear regression was conducted to determine if dominance and chin spot size were correlated and a positive linear relationship was found to exist between the two variables.     

Dynamics of Parental Behavior in the Black-Chinned Mouthbreeder,Tilapia melanotheron1 (Pisces: Cichlidae)2,3

Nur einige der Verhaltensweisen von Tilapia macrocephala (= melanotheron), die in der Brutpflegephase auftreten, werden davon beeinflußt, ob das ♂ Junge im Maul trägt. Alle Brutpflegehandlungen lassen sich von normalen Atmungs-, Körperpflege- und Nahrungsaufnahmebewegungen ableiten. Das Umschichten der Eier im Maul ist wahrscheinlich am wichtigsten. Es bewegt die Jungen im Atemwasserstrom und ändert sich — wie das Fächeln von Substratbrütern — mit dem Entwicklungsstand der Jungen. Es hängt aber auch noch von anderem ab: Umschichten stört, aber ersetzt auch funktionell das Atmen und hängt, wie dieses, mit dem Sauerstoffbedarf des Elterntieres zusammen. Vielleicht führt stärkere Sauerstoffzufuhr zu stärkerer Schwimmblasenfüllung. Diese muß kompensiert werden, ebenso das Gewicht der Jungen und ihr Gewichtsverlust, sobald ihre eigene Schwimmblase funktionsfähig ist. All das müßte das Atmen und Umschichten beeinflussen. Ein Maulbrüter muß Eier und Larven länger im Maul halten als ein Substratbrüter, muß also “Husten” und Schlucken entsprechend unterdrücken. Er muß ferner so gähnen, daß nichts aus dem Maul fällt; das Maulvorstülpen scheint ein derart modifiziertes Gähnen zu sein. Sich-Kratzen tritt während der ganzen Brutpflegezeit, aber nicht im Zusammenhang mit Ausschwimmversuchen der Jungen auf.     

Seasonal changes in the distribution of gonadal lipids and spermatogenetic tissue in the male phase of Monopterus albus (Pisces: Teleostei)

The gonad of Monopterus albus undergoes cyclical changes after the reversal of sex from female to male. The seasonally variable events include a prenuptial accumulation of cholesterol-positive lipid droplets in the cytoplasm of the interstitial cells when spermatogenetic activity is resumed in late February and early March. The development of the interstitial Leydig cells reaches a maximum in May just before spawning. There occurs a sudden depletion of the interstitial lipids during the breeding season in June at a time when the male animals exhibit active nuptial behaviour. After spermiation, the old interstitial cells degenerate, and during the succeeding phase of gonadal inactivity, become replaced by a new generation from connective tissue cells in the interstitium of the gonadal lamellae which gradually accumulate lipoidal material.
The lobular cycle comprises a postnuptial accumulation of amorphous intralobular lipids which become completely cleared in February when active spermatogenesis is restored. Spermatogenesis resumes shortly after spawning, but only advances as far as primary spermatocytes during the postnuptial period of inactivity.
The authors conclude that, as far as the seasonal variations in gonadal lipid distribution is concerned, the cycles in the gonad of the hermaphroditic teleost, M. albus , conform to the same pattern as those of the gonochoristic seasonal breeders studied.     

Paternal defence,cannibalism and polygamy: factors influencing the reproductive success of painted greenling (Pisces,Hexagrammidae)

The relative costs and benefits of paternal egg-guarding behaviour were identified for painted greenling, Oxylebius pictus, a small fish inhabiting temperate rock reefs. Because males must guard embryos against conspecifics of both sexes and against other fishes, their movements were curtailed, and an energy cost resulted. The males' somatic condition declined in proportion to the time spent guarding. Paternal males partially compensated by cannibalizing a small fraction of their progeny. Male O. pictus thus parasitize females by consuming their spawn. Polygamous matings were nonetheless advantageous for individual females as well as for males. The reproductive success of males and females increased for male broods with greater numbers of clutches, because there was a general dilution of predation loss in larger broods. The success of females, moreover, varied depending on the rank (chronological) order of spawning; success was greater for those that contributed earliest to the male's brood. The results support the hypothesis that the breeding cycles typical of paternal teleosts have evolved partly because of energy costs imposed by site-dependent care. For multiple-clutch species with protracted breeding seasons like O. pictus, the long-term benefits of polygamous spawnings perhaps outweight short-term losses sufficiently for filial cannibalism to evolve and be maintained. Alternatively (or in addition), the foraging milieu may exert a stronger average environmental influence than kin selection does.     

Phylogeographic patterns in populations of the black-chinned tilapia complex (Teleostei,Cichlidae) from coastal areas in West Africa: support for the refuge zone theory

Within the tilapiines, a major African cichlid lineage, quite limited genetic informations are available on phylogenetic interrelationships and phylogeographical patterns at both macro- and micro-evolutionary scales. The present study examines the genetic diversity of the black-chinned tilapia complex from coastal areas in West Africa (Senegal to Congo-Brazzaville) based on mtDNA control region sequences. Phylogenetic inferences provide support for the monophyly of both taxa involved. It is suggested that Sarotherodon melanotheron and Sarotherodon nigripinnis have diverged probably since the early Pleistocene. The occurrence of the main intraspecific lineages could be dated back to about 500,000-900,000 years. Our data suggest that West African Pleistocene refuge zones may have strongly influenced the degree and level of genetic differentiation among populations and thus the observed phylogeographic patterns. The spatial distribution of mtDNA lineages and the centres of intrapopulational genetic diversity clearly correspond to proposed lowland refuge zones and core areas of Central Africa. Moreover, this study genetically supports the existence of Sarotherodon melanotheron leonensis for the first time and further confirms the validity of the most recently introduced subspecies, e.g., Sarotherodon nigripinnis dolloi.     

Bone development in the jaw of Nile tilapia Oreochromis niloticus (Pisces: Cichlidae)

East African cichlids have evolved feeding apparatus morphologies adapted to their diverse feeding behaviors. The evolution of the oral jaw morphologies is accomplished by the diversity of bone formation during development. To further understand this evolutionary process, we examined the skeletal elements of the jaw and their temporal and sequential emergence, categorized by developmental stages, using the Nile tilapia Oreochromis niloticus as a model cichlid. We found that chondrogenesis started in Stage 17. The deposition of osteoid for the dermal bones commenced in Stage 18. The uptake of calcium dramatically shifted from the surface of larvae to the gills in Stage 20. The bone mineralization of the skeleton began in Stage 25. These data provide important information regarding the sequential events of craniofacial development in East African cichlids and lay the groundwork for studying the molecular mechanisms underlying adaptation of jaw structure to feeding behavior.     

The effects of early and late androgen treatments on the behavior of Sarotherodon mossambicus (Pisces: Cichlidae)

Groups of Sarotherodon mossambicus were treated with androgen by immersion or oral treatment at various stages of development. Fish were allowed to mature and the effects of treatment on gonadal and behavioral differentiation were examined. The effects of treatment on gonadal differentiation were assessed by determining the sex ratio for each group. Three treatments resulted in sex ratios significantly different from the 1:1 sex ratio obtained in untreated fish. Behavioral differences were detected between groups of males in three measures of territorial defense and aggression, and differences were detected between groups of females in two measures of male-female courtship interaction. A second experiment determined that early-treated females were more sensitive to a second androgen treatment later in life than females not exposed to androgen during development. A number of sex-reversed genetic females functioning as males were detected in two treatment groups with predominantly male sex ratios. There were no differences in the behavior of sex-reversed and non-sex-reversed male fish from the same treatment group. This study establishes that hormone treatments administered during development influence behavioral differentiation in a teleost.     

Development of the islets, exocrine pancreas, and related ducts in the Nile tilapia, Oreochromis niloticus (Pisces: Cichlidae)

Pancreatic development and the relationship of the islets with the pancreatic, hepatic, and bile ducts were studied in the Nile tilapia, Oreochromis niloticus, from hatching to the onset of maturity at 7 months. The number of islets formed during development was counted, using either serial sections or dithizone staining of isolated islets. There was a general increase in islet number with both age and size. Tilapia housed in individual tanks grew more quickly and had more islets than siblings of the same age left in crowded conditions. The pancreas is a compact organ in early development, and at 1 day posthatch (dph) a single principal islet, positive for all hormones tested (insulin, SST-14, SST-28, glucagon, and PYY), is partially surrounded by exocrine pancreas. However, the exocrine pancreas becomes more disseminated in older fish, following blood vessels along the mesenteries and entering the liver to form a hepatopancreas. The epithelium of the pancreatic duct system from the intercalated ducts to the main duct entering the duodenum was positive for glucagon and SST-14 in 8 and 16 dph tilapia. Individual insulin-immunopositive cells were found in one specimen. At this early stage in development, therefore, the pancreatic duct epithelial cells appear to be pluripotent and may give rise to the small islets found near the pancreatic ducts in 16-37 dph tilapia. Glucagon, SST-14, and some PPY-positive enteroendocrine cells were present in the intestine of the 8 dph larva and in the first part of the intestine of the 16 dph juvenile. Glucagon and SST-14-positive inclusions were found in the apical cytoplasm of the mid-gut epithelium of the 16 dph tilapia. These hormones may have been absorbed from the gut lumen, since they are produced in both the pancreatic ducts and the enteroendocrine cells. At least three hepatic ducts join the cystic duct to form the bile duct, which runs alongside the pancreatic duct to the duodenum.     

Annual gonadal cycle and reproduction in Patagonotothen tessellata (Richardson 1845 (Nototheniidae: Pisces) from the Beagle Channel, Argentina

The reproductive biology of Patagonotothen tessellata (Richardson 1845) in the Beagle Channel, Tierra del Fuego, Argentina was analysed from weekly captures over a 3-year period (1987–1989). The spawning periods and lengths thereof were analysed using GSI curves and monthly percentage distributions of maturation stages. Two gonadal maturation cycles and two spawning periods (winter and summer) within 1 year were defined. Also discussed is whether the same specimens are able to mate successfully in winter and summer, and whether the reproductive effort varies between the seasons. Male parental behaviour and the likely consequences on their energy levels are considered.