Effects of temperature manipulation on gamete development and reproductive activity in the farmed catfish Lophiosilurus alexandri.

Temperature is a critical factor for fish management, especially during breeding season. This study evaluated gametogenesis and gonadal maturation in the catfish Lophiosilurus alexandri under farming conditions in two temperatures. The fish were divided in the following experimental groups: (S0) control group, before starting the experiment, (S1) 30 days at 23 °C, (S2) 70 days at 23 °C, (S3) 30 days at 27 °C, and (S4) 70 days at 27 °C. Gonad, liver, and blood samples were obtained to analyze reproductive and health parameters. In females, increased gonadosomatic index (GSI) was associated with a decreased proportion of perinucleolar follicles and increased diameter of vitellogenic follicles in S4. Postovulatory follicles were found only in fish kept at 27 °C, whereas atretic follicles were more frequent at 23 °C. In S4, females had higher hepatic levels of vitellogenin and lower levels of zona radiata proteins. In males, a lower proportion of spermatogonia was found at 27 °C (S4), whereas GSI and proportions of spermatocytes, spermatids, and spermatozoa were similar among the groups. Levels of cortisol were higher in females of group S3. Plasma 17ß-estradiol, 11-ketotestosterone and 17α-hydroxyprogesterone were significantly higher in S4. Hematocrit, erythrocytes, and glucose were higher at 27 °C. Together, the present study indicate that exposure of L. alexandri to 27 °C provides suitable conditions for germ cell development and gonadal maturation in captivity, promotes increase of sex steroids, follicular growth, and oocyte maturation, with onset of ovulation occurring at 30 days. At 23 °C, spermatogonia proliferation is favored in males, but it cannot be recommended for stimulating female maturation of L. alexandri in fish farming.

Insulin-like growth factors 1 and 2 are associated with testicular germ cell proliferation and apoptosis during fish reproduction.

To support sperm production, fish testes undergo intense tissue remodelling, with endocrine, paracrine and autocrine signals regulating gonad physiology. The aim of this study was to investigate the testicular expression of insulin-like growth factor (Igf) 1 and Igf2 during spermatogenesis, and their relationship with cell proliferation and apoptosis throughout the reproductive cycle. The study was performed in male Hypostomus garmani, a catfish living in headwater rivers of the São Francisco River basin, Brazil. Spermatogenesis was analysed using histology, morphometry, immunohistochemistry and terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) analysis at different maturity stages. The results showed the proliferation of spermatogonia throughout the reproductive cycle, with a higher rate during the ripe stage. Germ and Sertoli cells expressed Igf1 at all stages of testicular maturity, Igf2 was predominant at the ripe stage and both Igf1 and Igf2 occurred at the spent stage. Caspase-3 and TUNEL analysis revealed a higher rate of apoptosis at the spent stage associated with reduced expression of Igf1 and Igf2. Sertoli cell proliferation was associated with spermatogonia and spermatocyte cysts at different stages of the reproductive cycle. Together, the data support a proliferative role for Igf1 and Igf2 in regulating testicular apoptosis in H. garmani, with cyclical variation in their expression during gonad maturation.

Expression of Vasa, Nanos2 and Sox9 during initial testicular development in Nile tilapia (Oreochromis niloticus) submitted to sex reversal.

Sexual differentiation and early gonadal development are critical events in vertebrate reproduction. In this study, the initial testis development and expression of the Vasa, Nanos2 and Sox9 proteins were examined in Nile tilapia Oreochromis niloticus submitted to induced sex reversal. To that end, 150O. niloticus larvae at 5 days post-hatching (dph) were kept in nurseries with no hormonal addition (control group) and 150 larvae were kept with feed containing 17α-methyltestosterone to induce male sex reversal (treated group). Morphological sexual differentiation of Nile tilapia occurred between 21 and 25 dph and sex reversal resulted in 94% males, whereas the control group presented 53% males. During sexual differentiation, gonocytes (Gon) were the predominant germ cells, which decreased and disappeared after that stage in both groups. Undifferentiated spermatogonia (Aund) were identified at 21 dph in the control group and at 23 dph in the treated group. Differentiated spermatogonia (Adiff) were found at 23 dph in both groups. Vasa and Nanos2 occurred in Gon, Aund and Adiff and there were no significant differences between groups. Vasa-labelled Adiff increased at 50 dph in both groups and Nanos2 presented a high proportion of labelled germ cells during sampling. Sertoli cells expressed Sox9 throughout the experiment and its expression was significantly greater during sexual differentiation in the control group. The results indicate that hormonal treatment did not alter initial testis development and expression of Vasa and Nanos2 in Nile tilapia, although lower expression of Sox9 and a delay in sexual differentiation was detected in the treated group.

Influence of low temperature on structure and dynamics of spermatogenesis during culture of Oreochromis niloticus.

Understanding the influence of different temperature conditions on the spermatogenesis is important for improvement of the fish aquaculture. This study evaluated the influence of low temperature on structural and quantitative dynamics of the spermatogenesis in Oreochromis niloticus. Adult males were cultivated with room temperature water (20.28-22.46°C) and testes were collected for histological, ultrastructural and morphometric analyses. This species has unrestricted lobular testis with cystic spermatogenesis and type I spermiogenesis that results in a single anacrosomal aquasperm. Seminiferous lobules and spermatogenic cells had a radial arrangement toward the spermatic duct. Superior and central portions of testes had a greater lobular area than the inferior portion in all samplings. Spermatogonia (9.3%) were distributed in the inferior portion of testes, spermatocytes (25.3%) and spermatids (34.4%) in the central portion, while spermatozoids (39.4%) and secretory cells (4.6%) in the superior portion. Throughout the study, correlation between water temperature and lobular area characteristics was significant only in the inferior portion of testes (r(2)=0.95), although the lobular area in the other testicular portions increased when the water temperature increased by 2°C. Correlation between the water temperature and spermatogenic cells was significant for undifferentiated spermatogonia (r(2)=0.54) and number of spermatids (r(2)=0.67). It is concluded that low cultivation temperatures may positively influence the generation of primary spermatogonia in the inferior periphery of O. niloticus testes. In addition, males maintain reservoirs of germ cells at low temperatures and the radial zonation of spermatogenesis has an important role in the renewal and production of germ cells.

PCNA and apoptosis during post-spawning ovarian remodeling in the teleost Oreochromis niloticus.

The balance between cell proliferation and apoptosis is crucial for tissue development and homeostasis. The present study investigated the contribution of proliferating cell nuclear antigen (PCNA) and apoptosis during ovarian remodeling after spawning in the Nile tilapia Oreochromis niloticus. Breeding females were kept in controlled conditions and ovary samples were collected weekly for TUNEL assay, immunohistochemistry for PCNA and caspase-3 and morphometric analysis. During the follicular growth, PCNA labeled mainly the nuclei of oocytes and follicular cells in a high proportion of follicles especially in primary growth, while a low occurrence of apoptosis in follicular and theca cells was detected. At 0-3 days post-spawning, post-ovulatory follicles showed no proliferative activity, however the follicular cells exhibited high rates of apoptosis. At 7-10 days, PCNA labeled the thecal cells in a low proportion of post-ovulatory follicles, which showed follicular cells with lower rates of apoptosis. PCNA labeled mainly the theca in the advanced and late stages of atretic follicles, while the follicular cells exhibited a significant increase of apoptosis along follicular atresia. We concluded that PCNA and apoptosis work cooperatively to ensuring the success of follicle development and maintaining of tissue homeostasis during follicular growth. PCNA and apoptosis are also essential mechanisms in the follicular regression during post-spawning ovarian recovery in the Nile tilapia.

Feeding and social behavior of the piabanha, Brycon devillei (Castelnau, 1855) (Characidae: Bryconinae) in the wild, with a note on following behavior

Knowledge concerning the behavior of wild freshwater fishes in Brazil is restricted to a few studies, despite such studies being able to answer fundamental questions about conservation. Species of Brycon are amongst the most threatened in the Neotropics, particularly in southeast Brazil, due to anthropogenic activities in this region. This study investigated the feeding and social behaviors of the endangered fish, Brycon devillei in the Preto River, Jequitinhonha basin, Minas Gerais State, Brazil. Behavioral data were collected by snorkeling with four spatially separated groups (habituated), and direct observations of shoals were made using an underwater video camera (a total of 448 hours of observations). This species showed diverse tactics to obtain food. However, the species proved to be predominately a specialist surface-picker, which adopted alternative tactics to find food at certain times of the year, most notably when food items on the water surface became low. Feeding frequency was shown to be negatively correlated to agonistic behavior between conspecifics. Feeding associations were also recorded between the anostomid Leporinus garmani, acting as a nuclear species, and B. devillei, as follower species. The data presented here showed the importance of conserving the riparian environment to protect B. devillei populations. Furthermore, the present study included rare observations of nuclear-follower feeding association among freshwater fishes, especially between medium-sized characiforms, being the first observations of such kind in a Cerrado stream.

O conhecimento do comportamento de vida livre em peixes de água doce brasileiros é restrito a poucos estudos e pode responder a questões fundamentais de conservação. Espécies do gênero Brycon estão entre as mais ameaçadas na região neotropical, especialmente no sudeste brasileiro, em vista de diversos impactos antrópicos. O presente estudo investigou os comportamentos alimentar e social de Brycon devillei no rio Preto, bacia do Jequitinhonha, Brasil. Para tanto, utilizou-se seções de mergulhos livres para observação direta de quatro grupos habituados de B. devillei, com o auxílio de câmeras filmadoras subaquáticas. Tal espécie apresentou variadas táticas para obtenção de alimento. Entretanto, a espécie demonstrou ser um forrageador de superfície especializado que, em determinados períodos do ano, quando provavelmente a lâmina d'água torna-se uma fonte escassa de recursos, adota táticas alternativas em busca de alimentos. A frequência de forrageamento demonstrou ser negativamente correlacionada com os comportamentos agonísticos. Foram observadas associações alimentares interespecíficas envolvendo o timburé, Leporinus garmani, como espécie nuclear, e a piabanha, Brycon devillei, como sua seguidora. Os dados apresentados no presente estudo demonstram a importância de se conservar o ambiente ripário em prol da proteção das populações de B. devillei. Além disso, o presente estudo é um dos raros registros de associação alimentar do tipo nuclear-seguidor entre peixes de água doce, especialmente entre espécies de médio porte, e o primeiro deste tipo para riachos de Cerrado.

Reproductive biology of the peacock bass Cichla piquiti (Perciformes: Cichlidae), an exotic species in a Neotropical reservoir

To analyze the reproductive biology of the peacock bass Cichla piquiti, 361 specimens were collected bimonthly in the Itumbiara Reservoir, southeast Brazil, from December 2004 to November 2005. Males and females in reproductive activity occurred during almost the entire year, with reproductive peak occurring before the beginning of the rains when the water temperature remained low, indicating that these environmental variables do not directly influence in the reproduction of C. piquiti. The long reproductive period, partially spent ovaries contained postovulatory follicles and oocytes in all developmental stages, indicate asynchronous development of oocytes and multiple spawning. The mean total lengthand body weigth were, respectively, 38.2 ± 7 cm and 965.0 ± 654.0 g for males and 37.4 ± 6.1 cm and 899.0 ± 495.0 g for females, statistically showing no sexual dimorphism in size. The smallest male and female found in advanced maturation stage measured 31.0 cm and 29.0 cm of total length, respectively. The body condition (K) of males and females did not present significant differences during the reproductive cycle and the slope (b) of the length-weight relationship was 3.22, suggesting that reproduction and the annual hydrology cycle do not interfere in the health condition. Cichla piquiti is an exotic piscivore fish that is well adapted to this Neotropical reservoir, which exhibits environmental conditions considerably different from its original habitat. This study indicates that the species presents plasticity in reproduction and in allocation of resources, probably due the aseasonality of the reservoir and the exploitation of native species.(AU)

Para analisar a biologia reprodutiva do tucunaré Cichla piquiti foram coletados bimestralmente 361 exemplares no reservatório de Itumbiara, sudeste do Brasil, no período de dezembro/2004 a novembro/2005. Machos e fêmeas em atividade reprodutiva ocorreram durante quase todo ano, com pico reprodutivo antes do início das chuvas, quando a temperatura da água permanecia ainda baixa, indicando que estas variáveis ambientais não influenciaram diretamente na reprodução de C. piquiti. O longo período reprodutivo, ovários parcialmente desovados contendo folículos pós-ovulatórios e ovócitos em todos os estádios de desenvolvimento, indicaram desenvolvimento assincrônico dos ovócitos e desova múltipla. A média de comprimento total e peso corporal foram, respectivamente, 38,2 ± 7,0 cm e 965,0 ± 654,0 g para machos e 37,4 ± 6,1 cm e 899,0 ± 495,0 g para fêmeas, estatisticamente não mostrando dimorfismo sexual em tamanho. Os menores exemplares, macho e fêmea, capturados no estádio de maturação avançada mediram 31,0 cm e 29,0 cm de comprimento total, respectivamente. O fator de condição (K) de machos e fêmeas não apresentou diferenças significativas durante o ciclo reprodutivo e o coeficiente angular (b) da relação peso-comprimento foi 3,22, sugerindo que a reprodução e o ciclo hidrológico anual não interferem na condição de saúde. Cichla piquiti é um peixe piscívoro exótico que está bem adaptado a este reservatório neotropical, o qual exibe condições ambientais consideravelmente diferentes de seu habitat original. Este estudo indica que a espécie apresenta plasticidade na reprodução e na alocação de recursos, provavelmente devido a não sazonalidade do reservatório e a exploração das espécies nativas(AU)