Germ cell survival and development critically depend on the cells' contact with Sertoli cells in the vertebrate testis. Fish and amphibians are different from mammals in that they show a cystic type of spermatogenesis in which a single germ cell clone is enclosed by and accompanied through the different stages of spermatogenesis by an accompanying group of Sertoli cells. We show that in maturing and adult testes from African catfish and Nile tilapia, Sertoli cell proliferation occurs primarily during spermatogonial proliferation, allowing the cyst-forming Sertoli cells to provide the increasing space required by the growing germ cell clone. In this regard, coincident with a dramatic increase in cyst volume and number of germ cells per cyst, in Nile tilapia, the number of Sertoli cells per cyst was strikingly increased from primary spermatogonia to spermatocyte cysts. In both African catfish and Nile tilapia, Sertoli cell proliferation is strongly reduced when germ cells have proceeded into meiosis, and stops in postmeiotic cysts. We conclude that Sertoli cell proliferation is the primary factor responsible for the increase in testis size and sperm production observed in teleost fish. In mammals, Sertoli cell proliferation in the adult testis is not observed under natural conditions. However, on the level of the individual spermatogenic cyst--similar to mammals--Sertoli cell proliferation ceases when germ cells have entered meiosis and when tight junctions are established between Sertoli cells. This suggests that fish are valid vertebrate models for studying Sertoli cell physiology.

Simple, effective protocols for cryopreserving milt of five Brazilian migratory characids (Brycon orbignyanus, Prochilodus lineatus, Piaractus mesopotamicus, Salminus maxillosus and Leporinus elongatus) and one Brazilian migratory catfish (Pseudoplatystoma corruscans) are described. Milt was frozen and stored in dry shippers immediately after collection, making the procedure practical for use in the field. A diluent of 5% glucose, 10% dimethyl sulphoxide and 10% egg yolk was effective for all of the characid species, and a diluent of 10% methanol and 15% powdered milk was effective for P. corruscans. Milt was thawed in an ambient temperature water bath (28–30° C). Activation of thawed milt was effective with either saline (0·9 or 0·45% NaCl) or bicarbonate (1% NaHCO3). Fertilization rates with frozen milt varied between 49 and 300% of the control. Application of the technology for increasing genetic diversity in hatchery programmes for enhancement, mitigation or conservation breeding is discussed, as is its application in aquaculture. Inappropriate use of the technology can threaten wild biodiversity and appropriate legislation and education may be required for its responsible use.

The Brazilian freshwater fish diversity is the richest in the world. Only 0.7% of all Brazilian species have had any aspect of their sperm biology addressed up to this date. The majority of the fish species described in this review migrate during the spawning season (a phenomenon known as piracema). Urbanization, pollution, hydroelectric dams and deforestation are some of the causes of stock depletion or even local extinction of some of these species. The knowledge concerning sperm quality and minimum sperm:egg ratio is important to maximize the use of males without reducing hatching rates. Furthermore, sperm cryopreservation and gene banking can guarantee the conservation of genetic diversity and development of adequate breeding programs of native fish species. In this review, we present and evaluate the existing information on Brazilian fish species that have been subject to sperm quality and cryopreservation studies. The following parameters were evaluated: volume of extractable sperm, sperm motility, sperm concentration, freezing media, freezing methods, and post-thaw sperm quality. Although the existing protocols yield relatively high post-thaw motility and fertilization rates, the use of cryopreserved sperm in routine hatchery production is still limited in Brazil.

Surubim, Pseudoplatystoma corruscans, is the most valuable commercial and recreational fish in the Sã o Francisco River, but little is known about adult migration and spawning. Movements of 24 females (9.5-29.0 kg), which were radio-tagged just downstream of Trê s Marias Dam (TMD) at river kilometer 2,109 and at Pirapora Rapids (PR) 129 km downstream of TMD, suggest the following conceptual model of adult female migration and spawning. The tagged surubims used only 274 km of the main stem downstream of TMD and two tributaries, the Velhas and Abaeté rivers. Migration style was dualistic with non-migratory (resident) and migratory fish. Pre-spawning females swam at ground speeds of up to 31 km day -1 in late September-December to pre-spawning staging sites located 0-11 km from the spawning ground. In the spawning season (November-March), prespawning females migrated back and forth from nearby pre-spawning staging sites to PR for short visits to spawn, mostly during floods. Multiple visits to the spawning site suggest surubim is a multiple spawner. Most post-spawning surubims left the spawning ground to forage elsewhere, but some stayed at the spawning site until the next spawning season. Post-spawning migrants swam up or downstream at ground speeds up to 29 km day -1 during January-March. Construction of proposed dams in the main stem and tributaries downstream of TMD will greatly reduce surubim abundance by blocking migrations and changing the river into reservoirs that eliminate riverine spawning and non-spawning habitats, and possibly, cause extirpation of populations.

The main objectives of the present study were to investigate the effects of 6-n-propyl-2-thiouracil (PTU) on Sertoli cell proliferation, germ cell number, and testis size in Nile tilapias (Oreochromis niloticus). In this regard, young fish (approximately 1 g BW and approximately 3.5 cm total in length) were treated for a period of 40 d with different concentrations (100 and 150 ppm) of PTU. The animals were killed and analyzed on d 1, 30, 40, 98, and 208 after the beginning of the treatment. On d 30 and 40 the spermatogenic process was delayed in fish treated with PTU compared with the control group. Also at these periods, treated tilapia had decreased (P < 0.05) body weight and total length. On d 98 body weight and total length had recovered in PTU-treated fish and were similar (P > 0.05) to those of the controls. However, testis weight and gonadosomatic index (testis mass/body weight) were approximately 100% higher (P < 0.05) in treated tilapia. Similarly, the area occupied by seminiferous tubules, the number of Sertoli cells and germ cells per cyst, and the number of Leydig cells per testis were significantly (P < 0.05) greater in treated fish. Nevertheless, nuclear volume and individual Leydig cell volume were significantly lower (P < 0.05) in tilapia receiving PTU treatment. Compared with controls, at 208 d all parameters analyzed presented the same trend as that observed at 98 d. In general, at 98 d the different PTU concentrations used during the treatment period induced similar effects. However, at 208 d the mean values observed for several parameters were significantly higher (P < 0.05) in fish exposed to 150 ppm. Probably due to the higher density of Sertoli cells per cyst in treated tilapia, these cells presented a smaller (P < 0.05) nucleolus and a trend to decrease its support capacity (efficiency). However, the meiotic index (germ cell loss during the two meiotic divisions) was similar (P > 0.05) in the three groups of fish investigated. Remarkably, the results found in tilapia were similar to those found for rats treated with PTU. This suggests strongly that the mechanisms of control of Sertoli cell and Leydig cell proliferation seem to be preserved during vertebrate evolution.

We used metadata on nine reproductive traits of 67 species of Brazilian iteroparous, oviparous, teleost freshwater fishes to test phenotypes associations to discriminate species that only spawn in large rivers (lotic fishes) from those capable to spawn in lentic habitats (lentic fishes). We tested the hypothesis that lotic fishes present spawning migration, shorter spawning season, single spawning, no parental care, free eggs, higher relative fecundity, faster embryogenesis, and larger size, while lentic fishes present no spawning migration, longer spawning season, multiple spawning, parental care, adhesive eggs, lower relative fecundity, slower embryogenesis, and smaller size. Our analyses supported the hypothesis but not all phenotypic associations satisfied it, specifically with regard to lentic fishes or to pairs of phenotypes typical of lentic fishes. We also concluded that spawning in large rivers is a better predictor of bionomic characters than spawning in lentic habitats, and lotic fishes are specialists compared to lentic fishes.

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