Sublethal effects of triclosan and triclocarban at environmental concentrations in silver catfish (Rhamdia quelen) embryos.

Although banished in some countries, triclosan (TCS) and triclocarban (TCC) have been detected in surface waters in concentrations ranging from ng L-1 to µg L-1 and have shown to affect non-target organisms posing risk to aquatic ecosystems. However, the majority of the studies consider higher levels of these chemicals and single exposure effects to investigate their potential risks, rather than using environmentally relevant concentrations and their binary mixture. In this study, the toxicity of TCS and TCC, and their binary mixture was assessed in catfish embryos (Rhamdia quelen, a south American native species) exposed to environmental concentrations during 96 h. Organisms were evaluated through the endpoints of developmental abnormalities (spine, fin, facial/cranial and thorax), biochemical biomarkers related to oxidative stress responses: catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) activities, protein carbonylation (PCO) and neurotoxicity by acetylcholinesterase activity (AChE). The data showed that TCS caused fin abnormalities, decrease of SOD activity and increase of AChE activity in the catfish embryos of 96hpf. On the other hand, TCC and the binary mixture showed a higher abnormality index for the 96hpf embryos, and an induction of CAT and GST activities for the mixture treatment. The results obtained were able to show potential, but not severe, toxicity of TCS and TCC even in low concentrations and a short period of exposure. The relevance of studies approaching real scenarios of exposure should be reinforced, considering environmental concentrations of chemicals, interactions of contaminants in complex mixtures and the use of a native species such as R. quelen exposed during initial stages of development.

Reproduction and fecundity of invader of internal insemination Trachelyopterus galeatus in a Neotropical floodplain

After the formation of Itaipu Reservoir, the invader Trachelyopterus galeatus colonized the upper Paraná River. Light microscopy was used to describe gametogenesis and the reproductive phases of females and males. The following data were verified: diameter of the oocytes, spawning type, batch fecundity by ovary weight, standard length, and total weight of the fish, along with the regions where this species reproduced in the upper Paraná River floodplain. A total of 470 specimens were collected quarterly in 2016, 2018 and 2019, and bimonthly in 2017. The gonads were fixed in a Karnovsky solution, dehydrated, infiltrated, and embedded in historesin. The histological slides were stained using PAS + iron hematoxylin + metanil yellow, analyzed and photographed under an image-capturing microscope. As regards diameter of the oocytes and fecundity estimates, ovaries whose oocytes were measured under a stereomicroscope were sampled. In the oogenesis, undifferentiated and differentiated oogonia, early primary growth oocytes, secondary growth oocytes, full-grown oocytes and maturing oocytes were recorded. In the spermatogenesis, primary and secondary spermatogonia, primary and secondary spermatocytes, spermatids and spermatozoa were recorded. The reproductive phases found for females and males were: immature, early development, late development, spawning/sperm-releasing capable, regression, and regeneration. Trachelyopterus galeatus prefers to occupy and reproduce in the Ventura, Patos, Guaraná, Fechada, Garças, and Pau Véio lagoons. The diameter of the oocytes varied from 0.4 to 2.9 mm. Females spawn, on average, 113 oocytes per batch. Batch fecundity variation shows that the larger the ovary, standard length, and total weight, the larger the number of oocytes to be spawned. This invader possesses reproductive success in the upper Paraná River floodplain, especially in lagoons.

Influence of light intensity and photoperiod on embryonic development, survival and growth of threatened catfish Ompok bimaculatus early larvae.

Larval growth and survival of catfishes are largely influenced by the various biotic and abiotic factors. The present study investigated the effect of different light intensities and photoperiods on growth and survival of Ompok bimaculatus larvae. Three separate trials of 21 days each were carried out in an aquarium tank. The first trial investigated the embryonic changes (based on hatching rate and time) upon exposure to varied light intensity (0, 300, 500, 900 and 1200 lx) and photoperiodic regime (24l:0d, 16l:8d, 12l:12d, 8l:16d and 0l:24d). Subsequently, hatched-out larvae were subjected to the aforementioned intensities (Trial II) and photoperiod (Trial III, intensity of 300 lx) for growth and survival attributes. Eight hundred healthy larvae (average body weight = 0.003 g) were randomly distributed into five treatment groups for the last two trials. Results suggest a higher embryo hatching rate and larval survival at 0 and 300 lx, whereas the largest larval growth was observed at 900 lx. In Trial III, survival was highest in 0l:24d and growth in 24l:0d and 16l:8d was higher (P < 0.05). Performance index was higher (P < 0.05) in both 0 and 300 lx light and decreased at higher intensities. The overall interpretation from the present study concludes that a completely dark rearing environment is recommended for better survival of O. bimaculatus although growth was compromised.

Cellular Invasion and Matrix Degradation, a Different Type of Matrix-Degrading Cells in the Cartilage of Catfish (Clarias gariepinus) and Japanese Quail Embryos (Coturnix coturnix japonica).

We previously studied the phenomena of the mesenchymal cell-dependent mode of cartilage growth in quail and catfish. Thus, we selected the two cartilage models in which mesenchymal cells participate in their growth. In such models, cartilage degradation occurred to facilitate cellular invasion. The studies do not explain the nature of the cartilage degrading cells. The current study aims to explore the nature of the cartilage-degrading cells using transmission electron microscopy (TEM) and immunohistochemistry. Samples of cartilage have been isolated from the air-breathing organ of catfish and the cartilage of the prospective occipital bone of quail embryos. Samples have been processed for TEM and immunohistochemistry. We found that two different cell types are involved in cartilage degradation; the macrophage in the cartilage of catfish and mesenchymal cells in the cartilage of the quail. Areas of cellular invasion in both catfish cartilage and quail embryo cartilage had an immunological affinity for MMP-9. In catfish, cartilage-degrading cells had identical morphological features of macrophages, whereas in quail embryos, cartilage-degrading cells were mesenchymal-like cells which had cell processes rich in vesicles and expressed CD117. Further study should consider the role of macrophage and mesenchymal cells during cartilage degradation. This could be valuable to be applied to remove the defective cartilage matrix formed in osteoarthritic patients to improve cartilage repair strategies.

Synergistic effect of water temperature and dissolved oxygen concentration on rates of fertilization, hatching and deformity of hybrid yellow catfish (Tachysurus fulvidraco♀×Pseudobagrus vachellii♂).

In the process of selecting and developing freshwater aquaculture species, yellow catfish (Tachysurus fulvidraco) have received widespread attention from Chinese farmers, fishery scientists and technologists. Achieving full artificial breeding of yellow catfish would help improve the quantity and quality of fingerlings supplied for large-scale production of this species. Temperature (T) and dissolved oxygen (DO) are the most important abiotic factors affecting the breeding efficiency of aquatic organisms. In this study, the synergistic effects of T and DO on fertilization rate (FR, %), hatching rate (HR, %) and deformity rate (DR, %) of hybrid yellow catfish (T. fulvidraco♀ × Pseudobagrus vachellii♂) were studied by central composite design (CCD) and response surface methodology. A quadratic regression model for the effects of T and DO on FR, HR and DR was established, and the combination of T and DO was optimized. The first and second order effects of T and DO on FR and HR were significant under the conditions of this experiment (P < 0.05). The first and second order effects of T on DR were significant (P < 0.05) but there was no significant effect of DO on DR (P > 0.05). T and DO had significant interaction effects on FR (P < 0.05). High T and high DO environments reduced FR and HR of yellow catfish eggs and increased DR of the newly hatched larvae. The optimal combination of T and DO was 26.0 °C and 8.3 mgL-1, respectively. Maximum FR and HR coincided with minimal DR whose predicted values were 87.2%, 89.1% and 2.7%, respectively, with reliability of 0.979. Maintaining T and DO in the best combination will help to improve breeding efficiency and ensure production of the highest quantity and quality of fingerlings.

Ontogeny of the catfish pectoral-fin spine (Teleostei: Siluriformes).

The characteristic and morphologically variable pectoral-fin spine of catfishes (order Siluriformes) has been well-investigated based on later developmental stages (juveniles and adults) but information on the earliest life stages are lacking. Here, we document the ontogeny of pectoral-fin spines in four siluroid (Ictalurus punctatus, Noturus gyrinus, Silurus glanis and Akysis vespa) and two loricarioid catfishes (Corydoras panda and Ancistrus sp.). To further our understanding of pectoral-fin spine development, we also examined adult and juvenile specimens representing 41 of the currently 43 recognized families of catfishes. Development of the pectoral-fin spine is similar in all catfishes and resembles the development of a typical soft fin ray. Fusion between hemitrichia of the anteriormost lepidotrichium occurs proximally first, forming the spine proper, with growth of the spine occurring through the subsequent fusion of developing distal hemitrichial segments that comprise the spurious ray. The variation of pectoral-fin spine morphology observed is largely attributed to the presence/absence of five traits, which either develop as part of the hemitrichial segments that are added to the distal tip of the spine during growth (distal rami, anterior/posterior serrae) or develop independent of these segments (denticuli and odontodes).

Microcystin and pyriproxyfen are toxic to early stages of development in Rhamdia quelen: An experimental and modelling study.

The recent increase of freshwater eutrophication has favored cyanobacteria blooms and consequently the increase of toxins such as microcystin-LR in aquatic environments, but few is know about the associated effect of toxin and other compounds. Pyriproxyfen is an insecticide indicated by WHO (World Health Organization) to control Aedes aegypti mosquito (vector of Dengue, Chikungunya and Zika diseases), however, the effects are not well described to non-target species, such as fish. The early life stages (ELS) of fish are more sensitive to chemical stress due to higher metabolic rate, immature immune system and high superficial area/volume ratio. In the current study, ELS of R. quelen a Neotropical fish were exposed to environmentally realistic concentrations of microcystin (1, 10 and 100 µg L-1; M1, M2 and M3 groups, respectively) from an algal extract, pyriproxyfen (1 and 10 µg L-1, P1 and P2) and their association (co-exposure). The hatching, survival and larvae deformities were analyzed, and applied a mathematical model to evaluate the effects on the population size along further generations. Both compounds were toxic to embryos/larvae of fish, but the effects were more pronounced in M2, P1M2 and P2M1 for hatching and M2, P1M2, P2M1 and P1 for survival. Deformities prevailed in groups exposed to the chemicals at 48 hpf (hours post-fertilization) were suggestions of toxicological interaction in P1M2, P2M1 and P2M2 at 48 and 72 hpf. In 96 hpf, the levels of deformities were lower than in previous times. Model predicted population density over 100 years decreased to lower than 0.5 (50%) in all groups, except for P1M1, indicating risk of extinction. P1M2 had the worse results, followed by M2, P1M3 and P2M1. Cyanobacterial blooms can lead to microcystin-LR levels higher than M2 (10 µg L-1), and the suggestion of toxicological interaction with pyriproxyfen is relevant because both compounds may potentially coexist in aquatic environments. Finally, mathematical models may provide an ecological interpretation of the risk of exposure of fish.

Expression profiling of c-kit and its impact after esiRNA silencing during gonadal development in catfish.

Receptor, c-Kit is a member of a family of growth factor receptors that have tyrosine kinase activity, and are involved in the transduction of growth regulatory signals across plasma membrane by activation of its ligand, kitl/scf. The present study analyzed mRNA and protein expression profiles of c-kit in the gonads of catfish, Clarias gariepinus, using real time PCR, in situ hybridization and immunohistochemistry. Tissue distribution analysis revealed higher expression mainly in the catfish gonads. Ontogeny studies showed minimal expression during early developmental stages and highest during 50-75 days post hatch, and the dimorphic expression in gonads decreased gradually till adulthood, which might suggest an important role for this gene around later stages of sex differentiation and gonadal development. Expression of c-kit was analyzed at various phases of gonadal cycle in both male and female, which showed minimal expression during the resting phase, and higher expression during the pre-spawning phase in male compared to females. In vitro and in vivo induction using human chorionic gonadotropin elevated the expression of c-kit indicating the regulatory influence of hypothalamo-hypophyseal axis. In vivo transient gene silencing using c-kit-esiRNA in adult catfish during gonadal recrudescence showed a decrease in c-kit expression, which affected the expression levels of germ cell meiotic marker sycp3, as well as several factors and steroidogenic enzyme genes that are involved in germ cell development. Decrease in the levels of 11-ketotestosterone and testosterone in serum were also observed after esiRNA silencing. The findings suggests that c-kit has an important role in the process of germ cell proliferation, development and maturation during gonadal development and recrudescence in catfish.

Synchronizing developmental stages in Neotropical catfishes for application in germ cell transplantation.

SummaryThe aim of this study was to describe the effect of temperature on the fertilization, early developmental stages, and survival rate of two Neotropical catfishes Pimelodus maculatus and Pseudopimelodus mangurus. After fertilization, the eggs were incubated at 22°C, 26°C, and 30°C, which resulted in fertilization rates of 96.95 ± 1.79%, 98.74 ± 0.76%, and 98.44 ± 0.19% for P. maculatus and 96.10 ± 1.58%, 98.00 ± 0.63%, and 94.60 ± 2.09% for P. mangurus, respectively. For P. maculatus, hatching occurred after 22 h 30 min post-fertilization at 22°C, 16 h 30 min at 26°C, and 11 h 20 min at 30°C, and the hatching rates were 43.87 ± 7,46%, 57.57 ± 17.49%, and 53.63 ± 16.27%, respectively. For P. mangurus, hatching occurred after 28 h 30 min post-fertilization at 22°C and 17 h 30 min at 26°C with respective hatching rates of 45.4 ± 21.02% and 68.1 ± 12.67%. For this species, all embryos incubated at 30°C died before hatching. Additionally, for P. maculatus, the larvae from the lower (22°C) and higher temperatures (30°C) presented increased abnormality rates, as observed in the head, tail and yolk regions. The lowest abnormality rate was detected at 26°C, which was considered the optimal incubation temperature for both species. The developed protocol enables the manipulation of embryonic development, which is important for the application of reproductive biotechniques, including chimerism and chromosome-set manipulation. The data obtained here are also important for the surrogate propagation of this species as P. mangurus was recently categorized as an endangered fish species.

Testicular germ line cell identification, isolation, and transplantation in two North American catfish species.

Our aim was to transplant blue catfish germ line stem cells into blastulae of triploid channel catfish embryos to produce interspecific xenogenic catfish. The morphological structure of the gonads of blue catfish (Ictalurus furcatus) in ~ 90- to 100-day-old juveniles, two-year-old juveniles, and mature adults was studied histologically. Both oogonia (12-15 µm, diameter with distinct nucleus 7-8 µm diameter) and spermatogonia (12-15 µm, with distinct nucleus 6-7.5 µm diameter) were found in all ages of fish. The percentage of germ line stem cells was higher in younger blue catfish of both sexes. After the testicular tissue was trypsinized, a discontinuous density gradient centrifugation was performed using 70, 45, and 35% Percoll to enrich the percentage of spermatogonial stem cells (SSCs). Four distinct cell bands were generated after the centrifugation. It was estimated that 50% of the total cells in the top band were type A spermatogonia (diameter 12-15 µm) and type B spermatogonia (diameter 10-11 µm). Germ cells were confirmed with expression of vasa. Blastula-stage embryos of channel catfish (I. punctatus) were injected with freshly dissociated blue catfish testicular germ cells as donor cells for transplantation. Seventeen days after the transplantation, 33.3% of the triploid channel catfish fry were determined to be xenogenic catfish. This transplantation technique was efficient, and these xenogenic channel catfish need to be grown to maturity to verify their reproductive capacity and to verify that for the first time SSCs injected into blastulae were able to migrate to the genital ridge and colonize. These results open the possibility of artificially producing xenogenic channel catfish males that can produce blue catfish sperm and mate with normal channel catfish females naturally. The progeny would be all C × B hybrid catfish, and the efficiency of hybrid catfish production could be improved tremendously in the catfish industry.

Management proposal based on the timing of oral incubation of eggs and juveniles in the sentinel species Genidens genidens (Siluriformes: Ariidae) in a tropical estuary

Ariids such as Genidens genidens are particularly susceptible to overfishing because of their complex reproductive mechanisms, which result in slow replenishment of stocks. In this study, we obtained data about the reproductive ecology of the catfish G. genidens in the Guanabara Bay estuary, Rio de Janeiro, Brazil, providing supporting information for management of its fishery. The sex ratio, mean size of each sex at first sexual maturation, and female fecundity were determined and the reproductive period was characterized. The males:females ratio was 1:2. The fecundity is low and increases proportionally to the females' growth in total length and weight. The breeding season occurs during the warmest months of the year, with spawning extending from October through March and incubation from December through April. Sexual maturation is late, with males maturing at a larger size than females. Skipping spawning was also identified in females. Based on the reproductive biology, it is suggested that the catch be restricted to individuals larger than 20 cm, considering the L75 of the males, and that fishing be prohibited from December through March, to include the main period of spawning season by females and oropharyngeal incubation by males.(AU)

Ariídeos, como Genidens genidens, são particularmente suscetíveis à sobrepesca devido a seus complexos mecanismos reprodutivos, que resultam em uma reposição lenta dos estoques. Neste estudo, obtivemos dados sobre a ecologia reprodutiva do bagre G. genidens no estuário da Baía de Guanabara, Rio de Janeiro, Brasil, fornecendo subsídios para seu manejo pesqueiro. A proporção sexual, o tamanho médio de primeira maturação sexual de cada sexo e a fecundidade das fêmeas foram determinados e o período reprodutivo caracterizado. A proporção machos:fêmeas foi de ١:2. A fecundidade é baixa e aumenta proporcionalmente ao crescimento das fêmeas em comprimento total e no peso. O período reprodutivo ocorre durante os meses mais quentes do ano, com a desova se estendendo de outubro a março e a incubação de dezembro a abril. A maturação sexual é tardia, com machos amadurecendo em tamanho maior que as fêmeas. Também foi identificado nas fêmeas adiamento da desova. Com base na biologia reprodutiva, sugere-se que a captura seja restrita a indivíduos maiores que 20 cm, considerando a C75 dos machos, e que a pesca seja proibida de dezembro a março, para incluir o período principal de época de desova das fêmeas e incubação orofaríngea dos machos.(AU)

Description of amino acid and fatty acid content during initial development of Lophiosilurus alexandri (Siluriformes: Pseudopimelodidae), a carnivorous freshwater catfish

Samples of eggs, newly hatched larvae (NHL), and larvae at the end of the lecithotrophic period (eight days after hatching) (LPL) of Lophiosilurus alexandri were collected to determine the amino acid and fatty acid profiles. Crude protein did not change throughout initial development and the concentration of lipids was highest in NHL. The content of the indispensable amino acids (IAA) isoleucine, leucine, and valine decreased in LPL, while in eggs and NHL they remained high and similar in value. The dispensable amino acids (DAA), such as aspartic acid, tyrosine, and glycine, increased in LPL, while alanine decreased. The percentage of neutral lipids increased in LPL. The saturated fatty acid content decreased during ontogeny, while monounsaturated fatty acids decreased only in LPL. The polyunsaturated fatty acid content was highest in LP. Polar fatty acids were found in higher percentages in eggs and NHL, but lower in LPL. Saturated fatty acid content decreased during ontogenetic development, while that of monounsaturated fatty acids decreased only in LPL. Polyunsaturated fatty acid content was highest in LPL. Protein content was maintained during ontogenetic development, while amino acid classes experienced changes. Lophiosilurus alexandri preferentially uses saturated and monosaturated fatty acids as an energy source during its early development.(AU)

Amostras de ovos, larvas recém-eclodidas (NHL) e larvas no final do período lecitotrófico (oito dias após a incubação) (LPL) de Lophiosilurus alexandri foram coletadas para determinar os perfis de aminoácidos e ácidos graxos. A proteína bruta não alterou durante o desenvolvimento inicial e a concentração de lipídios foi maior na NHL. O conteúdo dos aminoácidos indispensáveis (IAA) isoleucina, leucina e valina diminuíram em LPL, enquanto nos ovos e NHL eles permaneceram com valores elevados e semelhantes entre si. Os aminoácidos dispensáveis (DAA), como ácido aspártico, tirosina e glicina, aumentaram em LPL, enquanto a alanina diminuiu. A porcentagem de lipídios neutros aumentou em LPL. O teor de ácidos graxos saturados diminuiu durante a ontogenia, enquanto os ácidos graxos monoinsaturados diminuíram apenas em LPL. O teor de ácidos graxos poliinsaturados foi maior em LPL. Os ácidos graxos polares foram encontrados em porcentagens mais altas em ovos e NHL, e menores em LPL. O teor de ácidos graxos saturados reduziu durante o desenvolvimento ontogenético, enquanto que os ácidos graxos monoinsaturados diminuíram apenas em LPL. O teor de ácidos graxos poliinsaturados foi maior em LPL. O conteúdo de proteínas foi mantido durante o desenvolvimento ontogenético, enquanto as classes de aminoácidos sofreram mudanças. Lophiosilurus alexandri usa preferencialmente ácidos graxos saturados e monossaturados como fonte de energia durante o desenvolvimento inicial.(AU)

Developmental toxicity and DNA damaging properties of silver nanoparticles in the catfish (Clarias gariepinus).

Although, silver nanoparticles (AgNPs) are used in many different products, little information is known about their toxicity in tropical fish embryos. Therefore, this study evaluated the developmental toxicity of waterborne silver nanoparticles in embryos of Clarias gariepinus. Embryos were treated with (0, 25, 50, 75ng/L silver nanoparticles) in water up to 144h postfertilization stage (PFS). Results revealed various morphological malformations including notochord curvature and edema. The mortality rate, malformations, and DNA fragmentation in embryos exposed to silver nanoparticles increased in a dose- and embryonic stage-dependent manner. The total antioxidant capacity and the activity of catalase in embryos exposed to 25ng/L silver nanoparticles were decreased significantly while the total antioxidant capacity and the activity of catalase were insignificantly increased with increasing concentrations in the embryos from 24 to 144 h-PFS exposed to 50 and 75ng/L silver nanoparticles. Lipid peroxidation values showed fluctuations with doses of silver nanoparticles. Histopathological lesions including severely distorted and wrinkled notochord were observed. The current data propose that the toxicity of silver nanoparticles in C. gariepinus embryos is caused by oxidative stress and genotoxicity.

Lipid classes and fatty acids composition of the roe of wild Silurus glanis from subalpine freshwater.

The fat content and the fatty acid composition of the roe of European catfish (Silurus glanis) wild specimens captured in the Lambro river basin (Como, Northern Italy) were investigated in order to assess their nutritional value. Total extracted lipid values on roe weight were 5.8-6.3%. Phospholipids (40.2-43.6%) and triacylglycerols (31.8-34.7%) were the most represented lipid classes. Relevant amount of cholesteryl esters (14.3-15.6%) were also detected. The main fatty acids were palmitic, oleic and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA). Total saturated fatty acids were 28.1-30.9%, monounsaturated fatty acids were 28.2-30.6%, and polyunsaturated fatty acids (PUFAs) were 26.7-29.1%. The good found content of phospholipids and ω3-polyunsaturated fatty acids encourage further investigation for the exploitation of Silurus glanis roe in food products and/or supplements.

Cloning, localization and differential expression of Neuropeptide-Y during early brain development and gonadal recrudescence in the catfish, Clarias gariepinus.

Neuropeptide-Y (NPY) has diverse physiological functions which are extensively studied in vertebrates. However, regulatory role of NPY in relation to brain ontogeny and recrudescence with reference to reproduction is less understood in fish. Present report for the first time evaluated the significance of NPY by transient esiRNA silencing and also analyzed its expression during brain development and gonadal recrudescence in the catfish, Clarias gariepinus. As a first step, full-length cDNA of NPY was cloned from adult catfish brain, which shared high homology with its counterparts from other teleosts upon phylogenetic analysis. Tissue distribution revealed dominant expression of NPY in brain and testis. NPY expression increased during brain development wherein the levels were higher in 100 and 150days post hatch females than the respective age-matched males. Seasonal cycle analysis showed high expression of NPY in brain during pre-spawning phase in comparison with other reproductive phases. Localization studies exhibited the presence of NPY, abundantly, in the regions of preoptic area, hypothalamus and pituitary. Transient silencing of NPY-esiRNA directly into the brain significantly decreased NPY expression in both the male and female brain of catfish which further resulted in significant decrease of transcripts of tryptophan hydroxylase 2, catfish gonadotropin-releasing hormone (cfGnRH), tyrosine hydroxylase and 3ß-hydroxysteroid dehydrogenase in brain and luteinizing hormone-ß/gonadotropin-II (lh-ß/GTH-II) in pituitary exhibiting its influence on gonadal axis. In addition, significant decrease of several ovary-related transcripts was observed in NPY-esiRNA silenced female catfish, indicating the plausible role of NPY in ovary through cfGnRH-GTH axis.

Embryonic development in Zungaro jahu.

The aim of this study was to characterize the embryonic development of Zungaro jahu, a fresh water teleostei commonly known as 'jaú'. Samples were collected at pre-determined times from oocyte release to larval hatching and analysed under light microscopy, transmission electron microscopy and scanning electron microscopy. At the first collection times, the oocytes and eggs were spherical and yellowish, with an evident micropyle. Embryo development took place at 29.4 ± 1.5°C and was divided into seven stages: zygote, cleavage, morula, blastula, gastrula, organogenesis, and hatching. The differentiation of the animal and vegetative poles occured during the zygote stage, at 10 min post-fertilization (mpf), leading to the development of the egg cell at 15 mpf. From 20 to 75 mpf, successive cleavages resulted in the formation of 2, 4, 8, 16, 32 and 64 blastomeres. The morula stage was observed between 90 and 105 mpf, and the blastula and gastrula stage at 120 and 180 mpf; respectively. The end of the gastrula stage was characterized by the presence of the yolk plug at 360 mpf. Organogenesis followed, with differentiation of the cephalic and caudal regions, elongation of the embryo by the cephalo-caudal axis, and somitogenesis. Hatching occurred at 780 mpf, with mean larval total length of 3.79 ± 0.11 mm.

Molecular cloning, expression analysis and transcript localization of testicular orphan nuclear receptor 2 in the male catfish, Clarias batrachus.

Testicular receptor 2 (TR2; also known as Nr2c1) is one of the first orphan nuclear receptors identified and known to regulate various physiological process with or without any ligand. In this study, we report the cloning of full length nr2c1 and its expression analysis during gonadal development, seasonal testicular cycle and after human chorionic gonadotropin (hCG) induction. In addition, in situ hybridization (ISH) was performed to localize nr2c1 transcripts in adult testis and whole catfish (1day post hatch). Tissue distribution and gonadal ontogeny studies revealed high expression of nr2c1 in developing and adult testis. Early embryonic stage-wise expression of nr2c1 seems to emphasize its importance in cellular differentiation and development. Substantial expression of nr2c1 during pre-spawning phase and localization of nr2c1 transcripts in sperm/spermatids were observed. Significant upregulation after hCG induction indicate that nr2c1 is under the regulation of gonadotropins. Whole mount ISH analysis displayed nr2c1 expression in notochord indicating its role in normal vertebrate development. Taken together, our findings suggest that nr2c1 may have a plausible role in the testicular and embryonic development of catfish.

Characterization of the early development of Pseudoplatystoma reticulatum Eigenmann & Eigenmann, 1889 (Siluriformes: Pimelodidae) from the Paraguay River Basin

This study described the initial development of Pseudoplatystoma reticulatum, as well as changes in growth patterns. Morphometric and meristic variables were analyzed during embryonic, larval and juvenile periods, even as allometric growth coefficients for larvae and juveniles. Eggs showed an average diameter of 1.79 mm and broad perivitelline space (23.41%). The total length (TL) of the larvae ranged from 3.05 to 25.72 mm, and the total number of myomeres ranged from 45 to 50 (preanal = 13-16 and postanal = 30-35). Initial pigmentation is poor, concentrated at the extremities of the yolk sac. In the flexion stage, the pigmentation intensifies and forms a longitudinal stripe extending from the snout to the operculum and two longitudinal stripes on the body, one dorsal and the other ventral, that unite at the caudal peduncle. At first, there is a differential growth of the head and tail, and then there is the growth of the body, which shows changes in the ontogenetic priorities of feeding, swimming, and breathing capacities. Our results are very important for ecology, systematics, and hatchering, especially in terms of ontogenetic variation in morphology, growth, feeding, behavior, and mortality of P. reticulatum.

Este estudo descreveu o desenvolvimento inicial de Pseudoplatystoma reticulatum, bem como as mudanças nos padrões de crescimento, testando a hipótese da ocorrência de crescimento diferencial durante a ontogenia inicial da espécie. Variáveis morfométricas e merísticas foram analisadas durante os períodos embrionário, larval e juvenil, assim como os coeficientes de crescimento alométrico para larvas e juvenis. Os ovos apresentaram diâmetro médio de 1,79 mm e espaço perivitelino amplo (23,41%). O comprimento total (CT) das larvas variou de 3,05 a 125,72 mm e o número total de miômeros de 45 a 50 (pré-anal=13-16 e pós-anal=30-35). A pigmentação inicial é escassa, concentrada nas extremidades do saco vitelino. A partir do estágio de flexão a pigmentação se intensifica, formando uma faixa longitudinal que se estende do focinho ao opérculo e duas no corpo, uma dorsal e outra ventral, que se unem no pedúnculo caudal. Inicialmente há um crescimento diferenciado da cabeça e da cauda, com posterior crescimento do tronco, que demonstra mudanças nas prioridades ontogenéticas para as capacidades de alimentação, natação e respiração. Nossos resultados são de grande importância para a ecologia, sistemática e larvicultura, especialmente em termos de variação ontogenética na morfologia, crescimento, alimentação, comportamento e mortalidade P. reticulatum.

Gonadotropins and Growth Hormone Family Characterization in an Endangered Siluriform Species, Steindachneridion parahybae (Pimelodidae): Relationship With Annual Reproductive Cycle and Induced Spawning in Captivity.

The aim of this study was to identify and characterize pituitary cells of Steindachneridion parahybae females in captivity, highlighting the possible relationship with reproductive disorders at this level, since this species shows oocyte final maturation, ovulation and spawning dysfunction in captivity. The localization and distribution of growth hormone (GH), prolactin (PRL), somatolactin (SL), ß-luteinizing hormone (ß-LH), and ß-follicle stimulating hormone (ß-FSH) immunoreactive (-ir) cells in the adenohypophysis was studied by immunohistochemical and Western blot methods. In addition, cellular morphometric analyses and semi-quantification of ir-cells optical density (OD) during the annual reproductive cycle and after artificial induced spawning (AIS) were performed. Results showed that the distribution and general localization of pituitary cell types were similar to that of other teleost species. However, the morphometrical study of adenohypophysial cells showed differences along the reproductive cycle and following AIS. In general, females at the vitellogenic stage presented greater OD values for GH, PRL and SL than at other maturation stages (previtellogenic and regression stages), probably indicating an increased cellular activity during this stage. Conversely, ß-LH OD did not vary during the annual reproductive cycle. After AIS, ß-LH, SL and GH ir-cells showed an increase in OD values suggesting a possible involvement on oocyte final maturation, ovulation and spawning or a feedback control on the brain-pituitary-gonads axis. Reproductive dysfunction in S. parahybae females in captivity may be due to alteration of the synthesis pathways of ß-LH. In addition, GH family of hormones could modulate associated mechanisms that influence the reproductive status in this species.

Chilling of Steindachneridionparahybae (siluriformes: pimelodidae) embryos.

The objective of this study was to assess the viability of Steindachneridion parahybae embryos after chilling using different cryoprotectant solutions, stages of embryonic development, chilling curves, and storage periods at temperatures between -10 °C and 0 °C. Three experimental tests were conducted, and the following aspects were evaluated: (1) the toxicity of six cryoprotectant solutions (10% methanol, ethylene glycol, or DMSO combined with 0.5-M sucrose or lactose); (2) viability of embryos submitted to cooling with two cryoprotectant solutions (10% or 20% methanol combined with 10-M sucrose) at three different stages of development (closure of blastoporus, appearance of the optic vesicle and the moment when the tail began to straighten out), and two chilling periods (6 and 12 hours); (3) viability of embryos submitted to cooling with three chilling curves (directly to the freezer without a curve, 0.5 °C/min and 1.0 °C/min) and two chilling periods (6 and 12 hours). After the tests, it was concluded that the protocol which presented the most positive results after chilling, with a hatching rate of 63.50 ± 9.98% of the embryos and 12.32 ± 3.85% normal hatched larvae, was the one with embryos at the free-tail stage, the cryoprotectant solution with 10% methanol and 10-M sucrose, a chilling curve of 0.5 °C/min, stored for a maximum of 6 hours at subzero temperatures (temperature ranging between -5.05 °C and -7.83 °C).