Effects of dietary vitamin E on the growth and breeding performance of Ompok pabda

A 90 day feeding trail was conducted to investigate the effect of vitamin E on the growth and breeding performance of Ompok pabda. A total of 84 healthy female brood fish (41.10±0.44g) were divided into 4 treatments i.e. treatment T1 T2, T3 and T4 having three replications each. The fish were fed twice a day with a standard feed (40% protein) having 4 doses of vitamin E viz. 0 (served as control), 50, 100 and 150mg vitamin E/kg feed. At the end of the feeding trial, the brood fish were induced to breed with PG extract to observe the effect of vitamin Eon feed. After rearing for 90 days with the experimental feeds, it was found that weight gain and specific growth rate of brood fish fed with 100mg vitamin E/kg feed (treatment T3) was the highest (14.78±0.38g and 2.99±0.11) while 150mg vitamin E/kg feed (treatment T4) fed fish gave the poorest result (2.97±0.89g and 1.21±0.32). There was no significant difference in terms of length gain of brood fish among the different treatments. The brood fish were induced to breed with equal dose of PG extract (18 and 12mg PG/kg body weight for female and male respectively) to observe the dietary effect of vitamin E on breeding performance. The highest ovulation, fertilization and hatching rate of eggs were found to be 81.48±6.41%, 84.04±3.53% and 68.59±5.03% respectively in the brood fish of treatment T1 while the poorest (33.33±00%, 52.35±5.02% and 45.70±7.24% respectively) were found in the brood fish under treatment T4. The results suggest that inclusion of 100mg vitamin E/kg feed is best for enhancing the breeding performance of O. pabda brood fish indicating that vitamin E content has a positive impact on reproduction of fish. The present results also imply that inclusion of higher level of vitamin E exerts an antagonistic effect in terms of growth and breeding performance of this species.

In vivo and in vitro storage of kutum, Rutilus frisii kutum eggs

Effects of post-ovulatory and post-stripping retention time and temperature on egg viability rates were studied in kutum (Rutilus frisii kutum). Eggs were retained inside (in vivo storage) or outside the ovarian cavity with ovarian fluid (in vitro storage) at various temperatures. Two experiments were performed: 1) Partial volumes of eggs were stripped and fertilized at 24- hour intervals for 96 hours post-ovulation (HPO) (at 11 °C) and at 12-hour intervals for 72 HPO (at 14 °C), and 2) stored eggs were fertilized after 0, 2, 4, 6, and 8 hours post-stripping (HPS) at temperatures of 4, 10, 12, and 26 °C. In the first experiment, the highest eyeing and hatching rates (76% and 60% at 11 °C; 81% and 71% at 14 °C) and the lowest eyed-egg mortalities (20% at 11 °C; 12% at 14 °C) occurred in the eggs fertilized immediately (0–24 HPO at 11 °C and 0–12 HPO at 14 °C) after ovulation. Egg viability, as shown by successful eyeing and hatching rates, was completely lost by 72–96 HPO at 11 °C, and 60–72 HPO at 14 °C. In the second experiment, the maximum eyeing (87%) and hatching (75%) rates of eggs took place at 0 HPS followed by 8 HPS (> 80% and > 70%, respectively) at 4 °C. As storage temperature increased, egg viability decreased: 80%, 70%, and 50% viable at 8 HPS at 4, 10, and 12 °C, respectively. The eggs stored at 26 °C lost their viability almost completely after 4 HPS. Eyed-egg mortality increased from 13% at 0 HPS to 48.2% at 4 HPS at 26°C. These results demonstrate that egg stripping should take place within 168 °C-hours after ovulation and that complete loss of viability of the eggs occurs by 672°C-hours after ovulation. The in vivo storage method is more effective compared to in vitro storage. Also successful in vitro storage of eggs can be used atleast within 8 hours at temperatures ranging from 4 to 12ºC.

Physiological and biochemical study of over-ripened oocyte in the cultivated Caspian brown trout (Salmo trutta caspius) and determination of egg quality markers

To determine the best time for egg stripping after ovulation and over-ripened oocyte in the Caspian brown trout (Salmo trutta caspius), the eggs were retained in the parental abdominal cavity for 40 days post-ovulation (DPO) at 7±0.6°C. Eggs were stripped every 10-day interval in 4 treatment and were fertilized with a pool of semen obtained from 8 males. Also, the physiology and biochemistry of the eggs and ovarian fluids were studied. Results showed that the level of eyed eggs and hatched alevins declined with over-ripening time: that is, the expected amounts (90.65 ± 6.28% for eyeing and 86.33 ± 6.82% for hatching) in newly ovulated eggs (0–10 DPO) decreased to 0.67 ± 1.34% and 0.49 ± 0.98%, respectively, in over-ripened eggs (30–40 DPO). However, larval abnormalities remained constant for 30-days after ovulation. During the course of oocyte over-ripening, the pH of the ovarian fluid significantly decreased and the concentration of glucose, protein, calcium, iron, and aspartate aminotransferase activity significantly increased. Moreover, the concentration of protein, triglycerides, and aspartate aminotransferase activity in the eggs also changed. In the newly ovulated egg, the yolk consisted of homogenous tissue and its perivitelline space diameter had no considerable differences. With over-ripening, the yolk became heterogeneous, while chorion diameter and micropyle did not change. The perivitelline space diameter varied among different areas. The present study demonstrated that the best time to take Caspian brown trout eggs after ovulation at 7± 0.6°C was up to 10 DPO. Among the studied parameters of the egg and ovarian fluid, egg quality was related to both ovarian fluid parameters (e.g., pH, protein, aspartate aminotransferase, glucose, cholesterol, triglycerides, calcium, iron) and egg parameters (e.g., cholesterol, triglycerides, iron, aspartate aminotransferase). Thus, these parameters can be used as a egg quality markers in this species.

Water budgeting studies on the hatchery and nursery rearing practices for the common carp, Cyprinus carpio (Linnaeus, 1758)

Aquaculture systems are an integral element of rural development and therefore should be environment friendly as well as socially and economically designed. From the economic standpoint, one of the major constraints for the development of sustainable aquaculture includes externalities generated by competition in access to a limited resource. This study was conducted as an investigation into the water requirement for the hatchery and nursery production phases of common carp, Cyprinus carpio (Linnaeus, 1758) at the Maharashtra State Fish Seed Farm at Khopoli in Raigad Dist. of Maharashtra during the winter months from November to February. The water budgeting study involves the quantification of water used in every stage of production in hatchery and nursery systems and aimed at becoming a foundation for the minimization of water during production without affecting the yield; thereby conserving water and upholding the theme of sustainable aquaculture. The total water used in a single operation cycle was estimated to be 11,25,040 L [sic]. Out of the total water consumed, 4.74% water was used in the pre-operational management steps, 4.48% was consumed during breeding, 62.72% was consumed in the hatching phase, 21.50% was used for hatchery rearing and 6.56% was consumed during conditioning. In the nursery ponds, the water gain was primarily the regulated inflow coming through the irrigation channel. The total quantum of water used in the nursery rearing was 31,60,800 L [sic]. The initial filling and regulated inflow formed 42.60% and 57.40% respectively of water gain, while evaporation, seepage and discharge contributed 20.71%, 36.46% and 42.82% respectively to the water loss. The total water expended for the entire operation was 1,21,61,120 L [sic]. Water expense occurred to produce a single spawn in the hatchery system was calculated and found to be 0.56 L while the water expended to produce one fry was calculated as 4.86 L. The study fulfills the hydrological equation described by Winter (1981) and Boyd (1985). It also validates the water budget simulation model that can be used for forecasting water requirements for aquaculture ponds (Nath and Bolte, 1998).

Evaluate the efficiency of hydrogen peroxide and levaemisole hydrochloride in the treatment of Persian sturgeon, Chinese carp eggs and juveniles

During a two years research hydrogen peroxide efficacy evaluated for Persian sturgeon, Chinese carps and common carp eggs. These series of the experiments conducted in various conditions different concentration of hydrogen peroxide include 250, 500, 750, 1,000 1,500 2,000 3,000 and 9,000 PPM used as ten and fifteen minutes baths, compared with Malachite green and natural control . In the next phase effect of Levaemisole hydrochloride as an immunostimulator which applied as 5 mg/I in twenty minutes baths from day sixth after hatch evaluated by daily mortality rate and leukocytes counts. The results shown that according fertilization percent and temperature condition hydrogen peroxide at 1,000 and 1,500 PPM concentrations is a effective antifungal agent during incubation periods of Persian sturgeon and even sometimes increasing hatching rates significantly comparing with natural controls and Malachite green. In Chinese carps although hydrogen peroxide controls water molds but it is not recommended in high temperatures because it make shortened incubation time and mold infections will decrease. Also the results shown 750 PPM concentration of hydrogen peroxide in common carp eggs controls water moulds infections and increase hatching rate significantly comparing with Malachite green and natural control. Daily mortality rates accessing of Persian sturgeon fries show that 20 minutes baths of 5mg/1 levamisole hydrochloride decreases daily mortality rate during yolk sac absorption. Nitrogenous compounds: nitrate and ammonium differ significantly between treated tanks with control. Blood leucocytes concentrations as an immune index was different significantly in treated fishes by levamisole hydrochloride comparing with controls. In Chinese carps because yolks sac absorption time is short there is not necessary to use the levamisole hydrochloride. Although treated larvae were more active than controls. As a result our suggestions is to use hydrogen peroxide in Persian sturgeon and common carp artificial propagation and also suggest the use levamisole hydrochloride for Persian sturgeon beside management method in stress and pollution condition

Notes on the larval rearing scheme at SEAFDEC/AQD

For tiger shrimp, milkfish, and sea bass, larval rearing starts with the hatching of artificially spawned eggs. The eggs are stocked in larval rearing tanks, hatched, and metamorphosed larvae are fed and reared with good water management. Fry are harvested after about 30 days.

The diversity of macro invertebrates in the Victoria and Kyoga lake basin and their relationship to ecosystem functioning

Aquatic macro-invertebrates encompass all those organisms that be seen with unaided eyes. Most macro-invertebrates are categorised as semi-aquatic in that they are aquatic in early stages, but live as terrestrial organisms as adults, while others like gastropods, bivalves, Oligochaetae, Hirudinae and ostracods are exclusively aquatic. Some of them such as mayflies lay eggs in water and subsequent stages also live in water until adulthood when they emerge to live a terrestrial life. In others, eggs are laid near the water, while some like members of Tendipedidae (midges) lay their eggs on the leaves of aquatic macrophytes and after hatching their larvae creep into water

Observations on the biology of Nothobranchius guentheri (Pfeffer) (Cyprinodontidae), an annual fish from the coastal region of East Africa

Nothobranchius guntheri is found in seasonal pools and streams in the coastal region of Tanzania. A population recurring annually in a pond near Kilosa has been studied. Growth in length was rapid and maximum mean lengths were attained within 11-12 and 7-8 weeks of hatching by males and females respectively. Males grew larger and exhibited wider variation in length than females. N. guentheri shows clear sexual dichromatism. No significant inequality in the sex ratio was found. Females with ripe eggs were found 7-8 weeks after hatching. Spawning continued throughout adult life and fecundity increased markedly with increasing length. In laboratory aquaria, aggressiveness between adult males was noted and females were actively driven on to the substratum preparatory to spawning. The diet of the fish pond consisted chiefly of aquatic and terrestrial insects, of which midge larvae and pupae were the most common. N. guentheri is exploited by man in the aquarist trade and for the biological control of mosquitoes. An extended redescription of the species is appended which includes N. melanospilus (Pfeffer) as a synonym.

Studies on induced breeding of Clarias gariepinus (Burchell, 1822) in hapa pens

Induced breeding of Clarias gariepinus was conducted monthly in hapa pens, set up in Otamiri river for nineteen months (June 1993 - December 1994). Results of natural fertilization were unsatisfactory as few eggs were fertilized. Mean relative fecundity, percentage fertilization, percentage hatching and percentage fry survival were: 15.86 ± 1.95 x 10', 18.92 ± 5.28%, 13.50 ± 3.8% and 6.42 ± 0.72%. Results from artificial fertilization were as follows: Mean relative fecundity, 13.80 ± 2.85 x 10', percentage fertilization, 81.91 ± 2.28%, percentage hatching, 86.10 ± 2.46% and percentage fry survival, 21.40 ± 1.89% respectively. The success of artificial fertilization depended largely on the latency period of 9-11 hours and this suggests that induced breeding in pens is feasible. The poor results from natural fertilization were attributed to lack of adequate substrate for the male fish to display courtship and subsequent fertilization of eggs.

Ontogeny of IgM-producing cells in the mandarin fish Siniperca chuatsi identified by in situ hybridisation

The ontogeny of IgM-producing cells was studied in juvenile mandarin fish Simperca chuatsi, an important fish in China's aquaculture sector. The IgM-producing cells were localised through in situ hybridisation with a probe complementary to the Ig mu-chain in lymphoid-related tissues, including head kidney, spleen, thymus, intestine and gills. In head kidney, transcripts of Ig mu were first detected at 20 days post-hatching (dph) with a few positive signals. and the number of IgM-producing cells increased obviously from 39 dph onwards. At 136 dph, a large amount of positive cells were observed in the entire organ with clusters of these cells located around the blood vessels. In spleen, IgM-producing cells were found from 26 dph onwards, followed by an increase until 67 dph: clusters of positive cells were also detected around blood vessels at 102 dph. In thymus, IgM-producing cells were first observed at 39 dph; thereafter, no obvious increase was detected until 78 dph. The positive cells in thymus were distributed mainly in the outer zone of thymus. A few IgM-producing cells were still observed in thymus of 1-year-old mandarin fish. IgM-producing cells were not detected in the intestine until 87 dph, with several discrete positively stained cells distributed in the lamina propria. IgM-producing cells, scattered mainly in primary gill filaments around blood vessels, were detected in gills from 90 dph. As in other teleosts, these results indicated that the head kidney appears to be the primary organ for IgM production in mandarin fish, and IgM-producing cells exist in all organs examined in the present study, implying their lymphoid role in fish. In addition, it is suggested that vaccination after 20 dph may be much more effective in mandarin fish. (C) 2009 Elsevier B.V. All rights reserved.

Using drift nets to capture early life stages and monitor spawning of the Yangtze River Chinese sturgeon (Acipenser sinensis)

P>A sampling system for capturing sturgeon eggs using a D-shaped bottom anchored drift net was used to capture early life stages (ELS) of Chinese sturgeon, Acipenser sinensis, and monitor annual spawning success at Yichang on the Yangtze River, 1996-2004, before and just after the Three Gorges Dam began operation. Captured were 96 875 ELS (early life stages: eggs, yolk-sac larvae = eleuthero embryos, and larvae); most were eggs and only 2477 were yolk-sac larvae. Most ELS were captured in the main river channel and inside the bend at the Yichang spawning reach. Yolk-sac larvae were captured for a maximum of 3 days after hatching began, indicating quick dispersal downstream. The back-calculated day of egg fertilization over the eight years indicated a maximum spawning window of 23 days (20 October-10 November). Spawning in all years was restricted temporally, occurred mostly at night and during one or two spawning periods, each lasting several days. The brief temporal spawning window may reduce egg predation by opportunistic predators by flooding the river bottom with millions of eggs. During 1996-2002, the percentage of fertilized eggs in an annual 20-egg sample was between 63.5 to 94.1%; however, in 2003 the percentage fertilized was only 23.8%. This sudden decline may be related to the altered environmental conditions at Yichang caused by operation of the Three Gorges Dam. Further studies are needed to monitor spawning and changes in egg fertilization in this threatened population.

IgM, IgD and IgY and their expression pattern in the Chinese soft-shelled turtle Pelodiscus sinensis

Three Ig isotypes, IgM, IgD, and IgA, were previously known in reptiles. Here, in this report we describe IgM, IgD and a novel immunoglobulin heavy-chain isotype upsilon (IgY) in Chinese soft-shelled turtle (Pelodiscus sinensis). The IgM and IgY constant domains are characteristically similar to their counterparts described in other vertebrates. The expression of IgM and IgD were detected at mRNA level early during embryonic development, and their expression increased during further development. However, the IgY expression was not detected in larval turtles until 90 days after hatching-out. The increase in the transcription of these three Ig molecules was analyzed by using real-time PCR in spleen, kidney and blood following the injection of inactivated Aeromonas hydrophila. The primary increase in the expression of these three Igs was observed I week after the first injection, although not statistically significant, and the second injection 2 weeks after the first injection provoked a significant increase in the expression of these Igs, revealing a pattern of primary and secondary antibody response in the turtle. The present study represents the first report on reptile IgY and the pattern of IgM, IgD and IgY transcription in reptiles. (C) 2009 Elsevier Ltd. All rights reserved.

Predominant expression and cellular distribution of fish Agr2 in renal collecting system

Anterior gradient 2 (Agr2) genes encode secretory proteins, and play significant roles in anterior-posterior patterning and tumor metastasis. Agr2 transcripts were shown to display quite diverse tissue distribution in different species, and little was known about the cellular localization of Agr2 proteins. In this study, we identified an Agr2 homologue from gibe[ carp (Carassius auratus gibelio), and revealed the expression patterns and cellular localization during embryogenesis and in adult tissues. The full-length cDNA of CagAgr2 is 803 nucleotides (nt) with an open reading frame of 510 nt encoding 169 amino acids. The Agr2 C-terminus matches to the class I PDZ-interacting motif, suggesting that it might be a PDZ-binding protein. During embryogenesis, CagAgr2 was found to be transcribed in the mucus-secreting hatching gland from tailbud stage and later in the pharynx region, swim bladder and pronephric duct as revealed by RT-PCR and whole mount in situ hybridization. In the adult fish, its transcription was predominantly confined to the kidney, and lower transcription levels were also found in the intestine, ovary and gills. To further localize the Agr2 protein, the anti-CagAgr2 polyclonal antibody was produced and used for immunofluorescence observation. In agreement with mRNA expression data, the Agr2 protein was localized in the pronephric duct of 3dph larvae. In adult fish, Agr2 protein expression is confined to the renal collecting system with asymmetric distribution along the apical-basolateral axis. The data provided suggestive evidence that fish Agr2 might be involved in differentiation and secretory functions of kidney epithelium. (C) 2009 Elsevier Inc. All rights reserved.

Molecular cloning and expression characterization of ApoC-I in the orange-spotted grouper

Endogenous yolk nutrients are crucial for embryo and larval development in fish, but developmental behavior of the genes that control yolk utilization remains unknown. Apolipoproteins have been shown to play important roles in lipid transport and uptake through the circulation system. In this study, EcApoC-I, the first cloned ApoC-I in teleosts, has been screened from pituitary cDNA library of female orange-spotted grouper (Epinephelus coioides), and the deduced amino acid sequence shows 43.5% identity to one zebrafish (Danio rerio) hypothetical protein similar to ApoC-I, and 21.2%, 21.7%, 22.5%, 20%, and 22.5% identities to Apo C-I of human (Homo sapiens), house mouse (Mus musculus), common tree shrew (Tupaia glis), dog (Canis lupus familiaris) and hamadryas baboon (Papio hamadryas), respectively. Although the sequence identity is low, amphipathic alpha-helices with the potential to bind to lipid were predicted to exist in the EcApoC-I. RT-PCR analysis revealed that it was first transcribed in gastrula embryos and maintained a relatively stable expression level during the following embryogenesis. During embryonic and early larval development, a very high level of EcApoC-I expression was in the yolk syncytial layer, indicating that it plays a significant role in yolk degradation and transfers nutrition to the embryo and early larva. By the day 7 after hatching, EcApoC-I transcripts were observed in brain. In adult, EcApoC-I mRNA was detected abundantly in brain and gonad. In transitional gonads, the EcApoC-I expression is restricted to the germ cells. The data suggested that EcApoC-I might play an important role in brain and gonad morphogenesis and growth.

Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS

Perfluorooetanesulfonate (PFOS) is a persistent organic pollutant, the potential toxicity of which is causing great concern. In the present study, we employed zebrafish embryos to investigate the developmental toxicity of this compound. Four-hour post-fertilization (hpf) zebrafish embryos were exposed to 0.1, 0.5, 1, 3 and 5 mg/L PFOS. Hatching was delayed and hatching rates as well as larval survivorship, were significantly reduced after the embryos were exposed to 1, 3 and 5 mg/L PFOS until 132 hpf. The fry displayed gross developmental malformations, including epiboly deformities, hypopigmentation, yolk sac edema, tail and heart malformations and spinal curvature upon exposure to PFOS concentrations of I mg/L or greater. Growth (body length) was significantly reduced in the 3 and 5 mg/L PFOS-treated groups. To test whether developmental malformation was mediated via apoptosis, flow cytometry analysis of DNA content, acridine orange staining and TUNEL assay was used. These techniques indicated that more apoptotic cells were present in the PFOS-treated embryos than in the control embryos. Certain genes related to cell apoptosis, p53 and Bax, were both significantly up-regulated upon exposure to all the concentrations tested. In addition, we investigated the effects of PFOS on marker genes related to early thyroid development (hhex and pax8) and genes regulating the balance of androgens and estrogens (cyp19a and cyp19b). For thyroid development, the expression of hhex was significantly up-regulated at all concentrations tested, whereas pax8 expression was significantly up-regulated only upon exposure to lower concentrations of PFOS (0.1, 0.5, 1 mg/L). The expression of cyp19a and of cyp19b was significantly down-regulated at all exposure concentrations. The overall results indicated that zebrafish embryos constitute a reliable model for testing the developmental toxicity of PFOS, and the gene expression patterns in the embryos were able to reveal some potential mechanisms of developmental toxicity. (C) 2008 Elsevier Inc. All rights reserved.