Effect of cryoprotectants, extenders and freezing rates on the fertilization rate of frozen striped catfish, Pangasius hypophthalmus (Sauvage), sperm

The effects of four cryoprotectants (methanol, MeOH; dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; and ethylene glycol, EG), three extenders (calcium‐free Hanks' balanced salt solution, C‐F HBSS, Hanks' balanced salt solution, HBSS and sodium chloride, NaCl) and two different freezing procedures (one‐ and two‐step) on the cryopreservation of striped catfish (Pangasius hypophthalmus (Sauvage)) sperm were investigated. Sperm were frozen using a controlled‐rate freezer in 250 μL straws and stored for 2 weeks in a liquid nitrogen (LN₂) container. They were then airthawed at room temperature, and fertilization, motility and viability were assessed. The highest fertilization rate of 41% (81% of control) was achieved with the combination of 12% DMSO and 0.9% NaCl using a one‐step freezing procedure (10°C min^?1). Also, DMA resulted in a higher fertilization rate (30% or 51% of the control) than MeOH (18% or 38% of the control) or EG (8% or 12% of the control). In addition, the three extenders used did not affect fertilization rates after cryopreservation with each cryoprotectant. There were no significant differences among the three cryoprotectant concentrations and between the one‐ and two‐step freezing procedures. However, fertilization rates of cryopreserved sperm were significantly lower than the controls (P<0.05). The results of this study indicate that high fertilization rates of striped catfish eggs can be achieved using cryopreserved sperm when frozen at 10°C min^?1 in DMSO or DMA with either 0.9% NaCl or C‐F HBSS.     

Comparison of starch utilization in fingerlings of two Asian catfishes from the Mekong River (Pangasius bocourti Sauvage, 1880, Pangasius hypophthalmus Sauvage, 1878)

Five diets were formulated to provide an isoproteic daily dietary intake of 15 g kg^?1 day^?1, while maintaining daily starch intake ranging from 0 to 40 g kg^?1 day^?1. The 4‐week experiments started with initial mean weights of 4.7 and 4.4 g for Pangasius bocourti and P. hypophthalmus, respectively. The results clearly show the protein sparing action of starch in both species. Best protein retention was obtained with starch intake of 20 and 10 g kg^?1 day^?1 for P. bocourti and P. hypophthalmus, respectively, which was equivalent to 40 and 20% starch in the feed. Pangasius bocourti and P. hypophthalmus fingerlings attained maximal growth with starch intake of 30 and 10 g kg^?1 day^?1, equivalent to 60 and 20% starch in the feed. It was noted that body lipid accumulation was much higher in P. bocourti than in P. hypophthalmus, and that excess dietary starch intake depressed diet digestibility and growth.     

Effects of dietary protein and lipid content on growth performance and biological indices of iridescent Shark (Pangasius hypophthalmus, Sauvage 1878) fry

Dietary protein and lipid effects on growth, body composition and indices of iridescent Shark Pangasius hypophthalmus (Sauvage 1878) fry were studied using a 4 × 2 factorial design. Triplicate groups of 10 fish per tank, with initial mean weights of 3.54–3.85 g were fed eight isocaloric diets comprising a combination of four protein levels (250, 300, 350 and 400 g kg⁻¹ or 25%, 30%, 35% and 40%) and two lipid levels (60 and 120 g kg⁻¹ or 6% and 12%) respectively. The fish were hand-fed to satiety twice daily for 8 weeks. Specific growth rate (SGR) and feed conversion ratio (FCR) showed significant effects ( P <0.05) with variations in dietary protein and lipid. The highest SGR was observed in fish fed 40% protein/12% lipid diet but this value was not significantly ( P >0.05) different from the fish fed 30% protein/12% lipid diet. The FCR was lowest for the 40/12 diet and differed significantly only with the 25/6, 25/12 and 30/6 treatments respectively. The hepatosomatic index (HSI) was significantly affected by the level of protein, but intraperitoneal fat (IPF) showed significant variation due to dietary lipid level. The HSI significantly ( P <0.05) decreased when dietary protein increased from 25% to 30% but increased marginally thereafter. The IPF values increased with increased dietary lipid but decreased with increased dietary protein. Body protein was positively correlated with dietary protein content; conversely, body lipid content decreased with increase in dietary protein. The results of this experiment indicate the presence of a protein-sparing effect of lipid as fish fed 30% protein/12% lipid diet had growth and feed utilization comparable to those fed 40% protein/12% lipid diet.     

Growth,feed utilization and body composition of Asian catfish (Pangasius hypophthalmus) fed at different dietary protein and lipid levels

This study examined the effect of dietary protein and lipid levels on growth, feed utilization and body composition of Asian catfish Pangasius hypophthalmus reared in cages. Eight test diets were formulated at four protein (340, 380, 420 and 460 g kg⁻¹ crude protein) and two lipid (50 and 90 g kg⁻¹ crude lipid) levels. Fish (initial weight 4.7 g fish⁻¹) were fed the test diets for 8 weeks. Final body weight, weight gain (WG), feed intake (FI), feed conversion ratio (FCR), contents of crude protein, lipid and energy in whole body were dependent on both dietary protein and lipid levels, while specific growth rate (SGR), hepatosomatic index and body moisture content were dependent on dietary lipid level. The WG and SGR increased with the increase in either dietary protein level (at the same lipid level) or lipid level (at the same protein level). The FI and FCR decreased with the increase in dietary protein level (at the same lipid level) or lipid level (at the same protein level). Protein sparing action occurred in case dietary lipid level increased. Fish fed the diet containing 453 g kg⁻¹ crude protein and 86 g kg⁻¹ lipid had the highest WG and SGR, but the lowest FI and FCR, among the diet treatments. There were no significant differences in the protein retention efficiency (PRE) and energy retention efficiency (ERE) among the diet treatments, although PRE and ERE were relatively high in fish fed the diet containing 453 g kg⁻¹ crude protein and 86 g kg⁻¹ lipid. At the end of the feeding trial, body protein content increased, while body lipid content decreased, with the increase in dietary protein content at the same lipid level. Our results suggest that dietary levels of 450 g kg⁻¹ crude protein and 90 g kg⁻¹ lipid are adequate to support fast growth of P. hypophthalmus reared in cages.     

Cryopreservation of Mekong catfish,Pangasius bocourti Sauvage, 1880 spermatozoa

The effects of three extenders (Ginzburg fish ringer, Calcium‐free Hank's balanced salt solution, C‐F HBSS and sodium chloride, 0.9% NaCl) and four cryoprotectants (dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; methanol, MeOH and glycerol) in different concentrations (5%, 10% and 15%) on the motility, viability and fertilization rates of Mekong catfish (Pagasius bocourti) sperm were investigated. Sperm samples were transferred into 250‐μL French straws and sealed with a heated haemostat. The straws were then placed in a cryochamber. A computer‐controlled rate freezer (CL 3300) and programmable Cryogenesis, version 4 were used to regulate the freezing rate. The sperm samples were frozen at a rate of 10°C min^?1 from 4 to ?80°C and then evaluated after 72 h. Of the three extenders used with each cryoprotectant, C‐F HBSS had the highest fertilization rate of 75% (93% of control). This was not significantly different from the control treatment (fresh sperm) when tested with DMSO as the cryoprotectant. The lowest fertilization rate of 27% (38% of control) was resulting from the combination of 15% glycerol and C‐F HBSS. This study found that fertilization, motility and viability rates in all of the experiments had a positive significant correlation (P < 0.001).     

Effect of mixed feeding schedules with varying dietary protein levels on the growth of sutchi catfish, Pangasius hypophthalmus (Sauvage) with silver carp, Hypophthalmichthys molitrix (Valenciennes) in ponds

A 6‐month feeding trial was conducted in field condition using 10 farm ponds (400–600 m²) to investigate the effect of mixed feeding schedules on the growth of sutchi catfish, Pangasius hypophthalmus with silver carp, Hypophthalmichthys molitrix. Fish were stocked at a ratio of 80:20 (sutchi catfish, 4.9±0.5 g: silver carp, 12.0±0.8 g) at the total rate of 25 000 ha^?1. Two diets of high protein (30%, HP) and low protein (16%, LP) were prepared using locally available feed ingredients. Five different feeding schedules of high‐protein diet continuously (HP), low‐protein diet continuously (LP), 1‐day low–protein/1‐day high‐protein diet (1LP/1HP), 7 days low–protein/7 days high‐protein diet (7LP/7HP) and 14 days low–protein/14 days high‐protein diet (14LP/14HP) were tested. The fish were fed twice daily at the rate of 15%, 10%, 8% and 5% of their body weight for first, second, third month and rest of experimental period respectively. Feeding rate was calculated only on the basis of sutchi catfish weight only and was adjusted every 2 weeks according to weight gain. Fish fed LP and HP on alternate day (1LP/1HP) resulted in significantly (P<0.05) higher growth rate, feed utilization and production among the treatments. However, there were no significant differences (P>0.05) between the growth rates and production of fish fed HP regularly and fish fed 7 days LP followed by 7 days HP (7LP/7HP). Fish maintained on LP grew the least. The feed conversion ratio (FCR) values for sutchi catfish ranged between 2.04 and 2.79 with feeding schedule 1LP/1HP showing the best FCR. The total production of fish (including silver carp) ranged between 8310 and 12 422 kg ha^?1 6 months^?1 with 1LP/1HP feeding schedule resulting in the highest production and net profit. The study demonstrated that feeding fish continuously with HP is less economical. Thus, for profitable sutchi catfish culture with silver carp, farmers can use the mixed feeding schedule of alternate day feeding of LP and HP as a means of reducing feed costs.     

Fertilizability of cryopreserved and cadaveric fish spermatozoa of freshwater catfish Pangasius sutchi (Fowler, 1937)

Fertilizability of cryopreserved and cadaveric fish spermatozoa was attempted in the freshwater catfish Pangasius sutchi. Cryopreservation of spermatozoa was done with three cryoprotectants for short time storage (30 days). Whereas the spermatozoa obtained from the cadaveric fish were stored at ?20°C (30 days) without any cryoprotectants. Cryoprotectant toxicity assay showed maximum motility of 88.53 ± 2.01% and viability of spermatozoa (96.19 ± 4.92%) with 15% of Dimethyl acetamide (DMA) at 15 min equilibration time. Whereas Dimethyl sulfoxide (DMSO) (15%) registered moderate level of motility and viability 79.23 ± 2.02% and 80.89 ± 2.1%, respectively. However, the methanol (MeOH) (20%) resulted in low percentage of motility (58.6 ± 0.9%) and viability (68.6 ± 0.9%). Scanning electron micrographs further showed no significant deformity on the surface topography of spermatozoa of cadaveric fish as well as cryopreserved with DMA (15%). The results indicated that 15% of DMA with hanks balanced salt solution (HBSS) extender at a dilution ratio of 1:10 at ?80°C proved to be suitable for cryopreservation of spermatozoa in P. sutchi. This may be due to the osmolality of HBSS similar to seminal plasma of P. sutchi. Further studies on motility, viability and fertility potential of spermatozoa revealed 73.62 ± 1.61%, 88.34 ± 1.05% and 54 ± 2.2%, respectively, with DMA (15%). On the other hand, cadaveric fish sperm registered 57.12 ± 2.32%, 63.45 ± 0.94% and 25.33 ± 1.53% of motility, viability and fertilizability respectively. Thus, this study augments the feasibility of using cryopreserved as well as cadaveric fish spermatozoa for the seedling production in the fresh water catfish P. sutchi.     

Effects of different dietary lipid sources in the diet for Pangasius hypophthalmus (Sauvage, 1878) juvenile on growth performance,nutrient utilization,body indices and muscle and liver fatty acid composition

Four isonitrogenous (300 g kg^?1 crude protein), isoenergetic (21 kJ g^?1) experimental diets were formulated to contain fish oil (FO), soybean oil (SBO), crude palm oil (CPO) and linseed oil (LO), respectively, as lipid sources each at inclusion level of 120 g kg^?1 and fed to triplicate groups of 15 juvenile iridescent shark, Pangasius hypophthalmus (Sauvage, 1878) (mean weight 10.00 ± 0.70 g) to apparent satiation twice daily for 12 weeks. The results showed that survival of fish was consistently over 95% for all treatments whereas growth performance in the SBO and CPO treatments was similar and significantly (P < 0.05) higher than for fish fed the LO diet. However, fish fed all vegetable oil‐based diets performed better than those fed the FO diet. Muscle and liver fatty acid composition for all treatments generally reflected the composition in the diet and the ratio of n‐3/n‐6 was found to play an important role in P. hypophthalmus, suggesting that excessive amounts of n‐3 fatty acids reduce the overall growth performance. Results of this study thus suggests that P. hypophthalmus fed diets containing vegetable oils (especially CPO and SBO) produce better growth than those fed FO diet without showing any signs of nutrient deficiency.     

Anaesthetic efficacy of eugenol on iridescent shark,Pangasius hypophthalmus (Sauvage, 1878) in different size classes

Anaesthetic efficacy of eugenol was investigated on iridescent shark, Pangasius hypophthalmus. Fish (2, 5, 10 and 20 g) subjected to 20–200 mg L^?1 eugenol and behavioural response as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 27–300 s after exposure to 20–200 mg L^?1 eugenol, iridescent sharks reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 54–710 s exposure to such concentrations. Recovery time was 109–600 s in all weight classes as well as eugenol concentrations. Mortality (44–100%) was only observed in 2 g fish when subjected to 110–170 mg L^?1 eugenol. This study, for the first time, showed behavioural response of iridescent shark to anaesthesia as well as effectiveness of eugenol as anaesthetic in this important aquaculture‐ornamental species. According to the models obtained in this study, minimum eugenol concentrations to induce anaesthesia over less than 3 min were 53.8–81.5 mg L^?1 in 2–20 g fish. Likewise, maximum eugenol concentrations in which fish recovered over less than 5 min were 65.9–105.8 mg L^?1 in 2–20 g fish.     

Evaluation of Mixed-Feeding Schedules with Varying Dietary Protein Content on the Growth Performance and Reduction of Cost of Production for Sutchi Catfish,Pangasius hypophthalmus (Sauvage) with Silver Carp,Hypophthalmichthys molitrix (Valenciennes)

Abstract

An on-station pond trial was conducted for 6 months to investigate the suitability of mixed-feeding schedules with varying protein levels on the growth of sutchi catfish, Pangasius hypophthalmuswith silver carp, Hypophthalmicthys molitrixwith a view to minimizing the cost of production. Five different feeding regimes employed were: high protein diet (30% protein, H), low protein (15% protein, L), one day low protein diet followed by one day high protein diet (1L/1H), 7 day low protein diet followed by 7 day high protein diet (7L/7H), and 14 day low protein diet followed by 14 day high protein diet (14L/14H). Ten experimental ponds each of 0.008 ha size were divided into 5 treatment groups each with two replicates. Fishes were stocked at the ratio of 80:20 (sutchi catfish:silver carp) at the total rate of 25,000/ha. Feeding rate was calculated only on the basis of sutchi catfish weight only and fish were fed at the rate of 15,10, 8, and 5% of their body weight for 1st, 2nd, 3rd months, and the rest of the experimental period, respectively. The ranges of water quality parameters during the study period were: temperature 22°C to 32.8°C, dissolved oxygen 4.30 to 7.44 mg/L, pH 6.85 to 7.50, and transparency 21.9 to 32.2 cm. The result of the study showed that significantly (P < 0.05) higher fish weight gain was achieved in feeding schedules of H and 1L/1H and lowest with 14L/14H. The feed conversion ratio (FCR) values ranged between 2.06 and 2.51 with H and 1L/1H showing significantly the best FCR. The FCR values were very low when weight of silver carp was considered which ranged between 1.58 and 1.68. The survival (%) of sutchi catfish were lower (78.1 to 82.5%) compared to that of silver carp (90–95%). The total production of sutchi catfish ranged between 3594 and 7095 kg/ha and silver carp ranged between 1777 and 2220 kg/ha. A simple economic analysis showed that fish in 1L/1H fed alternately with low and high protein diets resulted in the highest net profit. The result of the study suggest that for profitable sutchi catfish farming in Bangladesh, farmers can adopt a mixed-feeding schedule of alternated low and high protein diet instead of continuously feeding a high protein diet.     

Establishing bacterial infectivity models in striped Catfish Pangasianodon hypophthalmus (Sauvage) with Edwardsiella ictaluri

A bacterial infectivity challenge model of Edwardsiella ictaluri in striped catfish was developed. All experiments were conducted using a bacterial isolate of E. ictaluri that had been recovered during a natural outbreak of bacillary necrosis of Pangasianodon (BNP) in farmed striped catfish Pangasianodon hypophthalmus in Vietnam. Time of immersion in 10⁷ CFU.ml⁻¹ had a significant effect on mortality. The immersion bacterial dose of 10⁷ CFU/ml for 30 s resulted in a cumulative percentage mortality of 63%. Three to four days post-bacterial challenge, fish showed gross clinical signs of natural BNP and E. ictaluri was recovered and identified from these fish. Moreover, a cohabitation challenge was evaluated as an alternative challenge method, although the mortalities among the infected fish were lower at around 15%–40%. This study confirmed the horizontal transmission of E. ictaluri in striped catfish and elucidated that cohabitation challenge could be used in reproducing the disease under controlled conditions.     

Heavy infection of Ichthyophthirius multifiliis in striped catfish (Pangasianodon hypophthalmus,Sauvage 1878) and its treatment trial by different therapeutic agents in a control environment

ABSTRACT

Striped catfish (Pangasianodon hypophthalmus) fingerlings were stocked (60 fishes/m³) in cemented square tanks for ornamental fishery purposes at the College Fish Farm in Mangalore. A total of 400 fishes with a mean weight (g) ranging from 3.24 ± 1.21 to 6.70 ± 1.13 and a mean length (cm) of 7.50 ± 0.94 to 9.50 ± 1.10 were examined. A severe outbreak of salt-like granule white spots was found on the body surface. Ichthyophthirius multifiliis, a ciliate pathogen, was identified as the causative agent by clinical signs, wet mount, and histopathological observations. Infected fishes were transferred and equally distributed to the 0.45 m³ glass aquaria and treated with three treatments: (T1) methylene blue + salt; (T2) raising temperature with salt; and (T3) formalin + malachite green. The best fingerling survival (55 ± 9.36%) was obtained by elevated water temperature with salt in T2.     

Options to reuse sludge from striped catfish (Pangasianodon hypophthalmus,Sauvage, 1878) ponds and recirculating systems

This paper compares the quantity and quality of solid waste obtained from traditional ponds and recirculating aquaculture systems (RAS) for striped catfish and evaluates methane and compost production from these wastes. Striped catfish sludge was collected from four commercial ponds along the Mekong river and from three indoor RAS. The amount of sludge dry matter produced per kilogram of fish in ponds was 6 times higher than that in RAS. However, the concentration of nutrients in solid waste from RAS was much higher, with better compost quality and higher methane yield than that of sludge from ponds. Out of the collected 381 L biogas, the methane yield of striped catfish’s solid waste in RAS systems was 201 L per kg chemical oxygen demand (COD). In ponds, the collected 267 L biogas yielded 125 L CH₄ per kg COD. The higher methane production from RAS sludge concurred with higher digestibility of COD: 58% for RAS versus 38% for ponds. The quality and quantity of methane from striped catfish sludge were lower than that for other animal manures, and the resulting electricity yield was low. Considering the higher nutrient concentration in RAS-sludge, we recommend the combination RAS and composting in reusing sludge, which is presently the best option for a more sustainable and cleaner striped catfish production system.     

Dietary ascorbic acid requirement for growth of striped catfish,Pangasianodon hypophthalmus (Sauvage, 1878) juveniles

A sixty‐day feeding trial was conducted to determine the ascorbic acid (AA) requirement for growth of striped catfish, Pangasianodon hypophthalmus juveniles. Seven iso‐nitrogenous and iso‐energetic (370 g protein per kg and 19.6 MJ/kg) purified diets were prepared with different levels of ascorbic acid such as control (0), T₁ (17.5), T₂ (35), T₃ (70), T₄ (175), T₅ (350) and T₆ (700) mg ascorbic acid (L‐ascorbyl‐2‐polyphosphate) equivalent per kg diet. Fish with a mean body weight of 3.2–3.4 g were stocked (fifteen fish per tank) in triplicates following a completely randomized design. Each group was fed to satiation twice a day for 60 days. Significant differences were observed in growth, survival, body composition and metabolic enzymes activities with different dietary ascorbic acid levels. Maximum weight gain, specific growth rate (SGR) and protein efficiency ratio (PER) were found in fishes fed with 35 mg AA per kg diet, supported by best feed conversion. Fish fed a diet containing vitamin C had the highest activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) compared to those fed with vitamin C‐depleted diets. In this study, based on using broken‐line regression analysis, the dietary vitamin C requirement for growth of P. hypophthalmus juveniles was estimated to be in the range of 46–76 mg AA per kg, depending on the criterion used, growth and liver storage. Our results will be helpful for the formulation of cost‐effective ascorbic acid incorporated diets for striped catfish, P. hypophthalmus.     

Oral administration of Escherichia coli lipopolysaccharide enhances the immune system of striped catfish,Pangasianodon hypophthalmus (Sauvage)

Lipopolysaccharide (LPS), a component of Gram negative bacteria, was reported as important immunostimulant for fish. In this study, striped catfish were fed diets containing different Escherichia coli LPS concentrations (0%, 0.01% and 0.05%) for 2 weeks and then fed control feed (0% LPS) for 4 weeks. Plasma cortisol and glucose were rather low and did not differ significantly among treatments (P > 0.05). The respiratory burst activity, lysozyme, complement, total of antibody as well as mortality in fish challenged with Edwardsiella ictaluri were recorded every 2 weeks (W2, W4 and W6). The lysozyme activity significantly increased in fish treated with LPS (P < 0.05) in W2, W4 and W6. The highest values of respiratory burst activity were observed at week 4 in fish fed 0.01% LPS. There were significant differences in total of antibody between fish fed LPS (0.01%) and control in W2, W4. The challenge test with Edwardsiella ictaluri showed that fish fed 0.01% LPS had lower cumulative mortality (40%, 33% and 42%) compared with the fish fed 0.05% LPS (50%, 40% and 47%) and control fish (40%, 57% and 53%) in the three difference sampling times respectively. These results suggest that feed supplemented with 0.01% LPS could enhance immunity of striped catfish after 2 weeks of oral administration and fish could be protected against bacterial infection during the following 4 weeks.