Experimental challenge studies in Vietnamese catfish,Pangasianodon hypophthalmus (Sauvage), exposed to Edwardsiella ictaluri and Aeromonas hydrophila

The two main diseases in the pangasius catfish industry are bacillary necrosis of Pangasianodon (BNP) and motile aeromonas septicaemia (MAS), where the aetiological agents have been identified as Edwardsiella ictaluri and Aeromonas hydrophila, respectively. In this study, apparently healthy Pangasianodon hypophthalmus were exposed to E. ictaluri, A. hydrophila or both bacterial species by intraperitoneal injection or immersion. There were 20 fish per treatment group, and the bacterial isolates used for the study were recovered from natural infections of BNP or MAS in farmed Vietnamese P. hypophthalmus. The results of the experimental infections mimicked the natural disease outbreaks reported from these pathogens in P. hypophthalmus. Furthermore, it was clearly demonstrated that E. ictaluri was only recovered from the fish exposed to the bacterium and not recovered from the animals receiving A. hydrophila.     

Intraspecific diversity of Edwardsiella ictaluri isolates from diseased freshwater catfish, Pangasianodon hypophthalmus (Sauvage), cultured in the Mekong Delta, Vietnam

A molecular epidemiology study was conducted on 90 Edwardsiella ictaluri isolates recovered from diseased farmed freshwater catfish, Pangasianodon hypophthalmus, cultured in the Mekong Delta, Vietnam. Thirteen isolates of E. ictaluri derived from diseased channel catfish, Ictalurus punctatus, cultured in the USA were included for comparison. All the E. ictaluri isolates tested were found to be biochemically indistinguishable. A repetitive (rep)‐PCR using the single (GTG)₅ primer was shown to possess limited discriminatory power, yielding two similar DNA profiles categorized as (GTG)₅‐PCR group 1 or 2 among the Vietnam isolates and (GTG)₅‐PCR group 1 within the USA isolates. Macrorestriction analysis identified 14 and 22 unique pulsotypes by XbaI and SpeI, respectively, among a subset of 59 E. ictaluri isolates. Numerical analysis of the combined macrorestriction profiles revealed three main groups: a distinct cluster formed exclusively of the USA isolates, and a major and minor cluster with outliers contained the Vietnam isolates. Antibiotic susceptibility and plasmid profiling supported the existence of the three groups. The results indicate that macrorestriction analysis may be regarded as a suitable typing method among the E. ictaluri species of limited intraspecific diversity. Furthermore, the findings suggest that E. ictaluri originating from Vietnam may constitute a distinct genetic group.     

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.     

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.     

Evaluation of soybean meal in the formulated diets for juvenile Pangasianodon hypophthalmus (Sauvage, 1878)

The influence of soybean meal (SBM) on the growth and feed utilization of juvenile sutchi catfish (Pangasianodon hypophthalmus, Sauvage, 1878) was investigated. Eight isonitrogenous (300 g kg^?1 CP) and isoenergetic (18 MJ kg^?1) diets were formulated incorporating Argentine SBM to replace fish meal at 0, 150, 300, 450, 600, 750, 900 and 1000 g kg^?1 dietary protein. Each diet was fed to three replicate groups of fish with an initial weight 6.0–6.2 g for twelve weeks. Growth performance decreased, and feed utilization was worsened with the increase in SBM inclusion in the diets. Final weight and relative growth rate (RGR) of fish fed control diet (0 SBM) were significantly higher than those fed test diets (P < 0.05). However, there were no significant differences in the final weight and RGR between the fish fed on 15 SBM, 30 SBM, 45 SBM and 60 SBM diets. Specific growth rate of fish fed 0 SBM, 15 SBM, 30 SBM and 45 SBM diets was significantly higher than those fed other diets. Feed conversion ratio of fish fed 0 SBM, 15 SBM, 30 SBM, 45 SBM and 60 SBM diets was significantly lower than those fed other diets (P < 0.05). The dry matter and protein digestibility were lesser in all the diets in comparison with the control diet. Hepatosomatic index and viscerosomatic index increased with increasing SBM in diet. This present trial indicated that fish meal can be replaced by SBM in the diet of juvenile sutchi catfish only up to 45% of fish meal protein without any adverse effect on growth, feed utilization and body composition.     

The economics of sutchi catfish (Pangasianodon hypophthalmus) aquaculture under three different farming systems in rural Bangladesh

This paper examines production costs and returns of sutchi catfish (Pangasianodon hypophthalmus) aquaculture under three different farming systems in Bangladesh. Based on the production technology, sutchi catfish farming is classified as extensive, semi‐intensive and intensive. Results showed that sutchi catfish farming is profitable irrespective of the level of intensification and in all three instances the cost of feed dominated the variable costs of production. The average annual production costs were estimated at US$5217 ha^?1 in intensive farming, while US$2694 ha^?1 in semi‐intensive and US$981 ha^?1 in extensive farming. Despite the higher production costs per hectare, the average annual net return was higher in intensive farming (US$3364), compared with semi‐intensive (US$2048) and extensive (US$1099) farming. The average annual production per hectare under intensive farming conditions (13 945 kg) was higher than semi‐intensive (7705 kg) and extensive (3380 kg) farming mainly due to higher levels of inputs, including seed, feed, fertilizer and labour. However, the Cobb–Douglas production function model suggests that inputs are inefficiently used in the intensive farming system. Conversely, there is enough scope to increase the production and income from the semi‐intensive and extensive farming systems by using more inputs.     

Breeding for improved resistance to Edwardsiella ictaluri in striped catfish (Pangasianodon hypophthalmus): Quantitative genetic parameters

Bacillary necrosis of Pangasius (BNP), caused by Edwardsiella ictaluri, is one of the most devastating diseases in striped catfish farming. To date, quantitative genetic inheritance of BNP resistance is not known in striped catfish Pangasianodon hypophthalmus. The main aim of this study was to estimate genetic parameters for BNP resistance in a breeding population of striped catfish undergoing four generations of selection for high growth. Specifically, the study examined whether BNP resistance is heritable to enable family selection and whether genetic improvement for enhanced BNP resistance may have detrimental effects on growth and survival rate. To test these hypotheses, 720 full‐ and half‐sib families were challenged with E. ictaluri pathogen using injection and cohabitation methods over four years, from 2010 to 2012 and 2015. In total, the data included 398,234 animals in the pedigree, from which 18,849 animals had disease challenge test records and 39,103 siblings had growth performance. Both univariate and bivariate sire–dam linear and threshold mixed models were used to estimate (co)variance components for BNP resistance, survivals and growth traits. The estimates of heritability for the BNP resistance recorded as death or survival were low regardless of models used (0.10–0.16), whereas survival time (days post‐challenge test) showed moderate heritability (0.35). The survival rate during hapa rearing had medium heritability (0.33–0.52). The genetic correlations of BNP resistance with body weight and survival were all positive (0.03–0.53), suggesting that selection of increased BNP resistance may have positive impacts on growth and survival traits, and these traits could be easily improved simultaneously in the selective breeding programme for striped catfish.     

Fertilization efficiency of cryopreserved sperm from striped catfish, Pangasius hypophthalmus (Sauvage)

The fertilization efficiency of cryopreserved sperm was compared with fresh sperm from striped catfish, Pangasius hypophthalmus . Of the two sets of experiments carried out, the first compared four sperm doses using fresh sperm and fresh eggs. The second experiment compared six concentrations of cryopreserved sperm ranging from 6.94 × 10⁷ to 6.94 × 10¹⁰ to fertilize 100 eggs per batch. Fertilization, hatch and survival rates were compared between cryopreserved and fresh sperm. The highest fertilization rate (53.75±1.62%) was achieved with a sperm dose of 6.94 × 10⁸. Increasing the sperm dose to 3.47 × 10⁹ did not increase the fertilization rate, indicating that the optimum sperm:egg ratio lies between 6.94 × 10⁶ and 3.47 × 10⁷ sperm per egg. Both highest (6.94 × 10¹⁰) and the lowest (6.94 × 10⁷) sperm doses resulted in lower fertilization rates (2.04% and 16.90% respectively). No significant differences were found among four fresh sperm doses compared. Mean hatch and survival rates resulting from fresh and cryopreserved sperm were similar. The experiment shows that while only 1.89 × 10⁶ fresh spermatozoa was required to fertilize a fresh egg, 6.94 × 10⁶ (or 3.67 times more) cryopreserved sperm was required to achieve the same level of fertilization. This provides important information for making decision to cryopreserve sperm for commercial and/or conservation purposes.     

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.     

Cross-protection elicited in channel catfish (Ictalurus punctatus Rafinesque) immunized with a low dose of virulent Edwardsiella ictaluri strains

Cross-protection of channel catfish ( Ictalurus punctatus Rafinesque) immunized with a low dose of virulent Edwardsiella ictaluri and challenged with six E. ictaluri strains was examined in four trials. The relative per cent survival among low-dose immunized and then challenged fish ranged from 27.7% to 100%. Significant protection ( P <0.05), with the exception of strain ATCC-33202, was conferred by immunization with a given E. ictaluri strain challenged either with a homologous or a heterologous strain. Antibody titres of pooled serum collected on day 22 from surviving fish examined by enzyme-linked immunosorbent assay (ELISA) ranged from 1:40 to 1:320, but no differences were apparent among different vaccinated groups. The protein profiles of six E. ictaluri strains examined by Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a relatively homogeneous pattern. Immuno-blots probed with pooled serum from immunized and challenged fish showed a pattern similar to LPS-reaction patterns observed with E. ictaluri in other studies. Since the present studies further corroborate that E. ictaluri is a clonal bacterial species with no apparent antigenic differences, it is possible that immunization with a single E. ictaluri field strain should confer protection against any other strain.     

Striped catfish (Pangasianodon hypophthalmus,Sauvage, 1878) aquaculture in Bangladesh: an overview

Farming of the striped catfish, Pangasianodon hypophthalmus, is a major aquaculture activity in Bangladesh, particularly in the district of Mymensingh. However, pangasius farm management practices and the socio‐economic impacts of pangasius farming systems in Mymensingh have not yet been adequately described in the literature. This article provides an overview of the present status and characteristics of pangasius culture in Bangladesh based on data from a study conducted in Mymensingh district during 2009. The mean productivity of pangasius was 36.9 MT ha^?1, where 87.9% of the farms produced between 15 and 65 MT ha^?1 of pangasius, with an additional 10–20% Indian major and Chinese carps and Nile tilapia. Pangasius aquaculture in Bangladesh has improved the economic and social status of a variety of stakeholders in communities where the fish is farmed.     

Isolation and characterization of Edwardsiella ictaluri strains as pathogens from diseased yellow catfish Pelteobagrus fulvidraco (Richardson) cultured in China

Yellow catfish Pelteobagrus fulvidraco (Richardson) is a commercially important fish generally distributed in Southeast Asian countries. The well‐known aetiological agent of enteric septicaemia of catfish, Edwardsiella ictaluri, was isolated from diseased yellow catfish P. fulvidraco (Richardson) reared at two commercial fisheries in China. The economic losses due to the high mortalities (about 50%) caused by this bacterium have been increasing annually. The affected fish presented two different, typical symptoms: pale gills, slight exophthalmia and a ‘hole in the head’, and haemorrhage on the opercula, in the skin under the jaw, creating a ‘hole under the jaw’. These diseases were found frequently in cultured yellow catfish throughout China. The isolates from both outbreaks were all Gram negative, facultatively anaerobic and short rod. Morphological and biochemical tests and phylogenetic analysis based on the 16S rDNA sequences all strongly indicated that these yellow catfish isolates were highly identical to the known E. ictaluri. In addition, the isolates possessed the typical plasmid profile of E. ictaluri. Experimental infection assays were conducted and pathogenicity (by an intraperitoneal injection) was demonstrated in yellow catfish and channel catfish Ictalurus punctatus. The results showed that yellow catfish isolates were quite conservative phenotypically and genetically, and were able to cause two different, typical symptoms in this fish under unknown conditions and mechanism.     

黄颡鱼鲇爱德华氏菌的分离鉴定及其致病性研究

从患头顶溃疡症的黄颡鱼(Pelteobagrus fulvidraco)内脏组织中分离出致病力强的菌株JZ086,对该菌进行了形态学、生理生化特性的测定,并进行了Biolog全自动微生物分析系统及16S rDNA序列鉴定。结果显示:人工感染实验证实该菌为黄颡鱼的病原菌,其半数致死量(LD50)为1.7×105CFU。Biolog全自动微生物分析系统鉴定结果显示该菌为鲇爱德华氏菌(Edwardsiella ictaluri)。16S rDNA序列分析显示该菌与鲇爱德华氏菌的亲缘关系最近,同源性高达99%;系统进化树中与鲇爱德华氏菌(AB050826)自然聚为一支,二者遗传距离约为0.002。鉴定结果确认JZ086为鲇爱德华氏菌。药物敏感性试验结果显示,该菌对所测试的15种药物都敏感,其中对氨苄西林、环丙沙星、左氧氟沙星、妥布霉素等尤其敏感。     

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.     

Susceptibility of channel catfish, Ictalurus punctatus (Rafinesque), to Edwardsiella ictaluri challenge following copper sulphate exposure

Channel catfish, Ictalurus punctatus (Rafinesque), with or without a preliminary 24 h exposure to 2 mg copper sulphate L(-1), were challenged with 7.5 x 10(6) colony forming units L(-1) of Edwardsiella ictaluri to determine the effect of copper sulphate on disease resistance. Catfish previously exposed to copper sulphate were significantly more resistant to the bacterial challenge than those not exposed. Catfish not exposed to copper sulphate suffered 35.5% mortality while catfish exposed to copper sulphate experienced 14.1% mortality. Copper concentrations were the same in tank waters of both exposed and control fish at the time of challenge, eliminating the possibility that copper in the water may have been toxic to bacteria. Copper concentrations in freeze dried and ground tissues of unexposed, exposed, and purged channel catfish were highest in fish before copper sulphate exposures suggesting that elevated tissue levels of copper were not responsible for the increased resistance to bacterial challenge. Competition for sites of bacterial attachment to gill or epithelial cells may account for the reduction in mortality; although this is not supported by the low copper content of fish tissue after copper exposure.     

A comparative study of Edwardsiella ictaluri parent (EILO) and E. ictaluri rifampicin-mutant (RE-33) isolates using lipopolysaccharides,outer membrane proteins,fatty acids,Biolog, API 20E and genomic analyses

The biological properties of Edwardsiella ictaluri RE-33 rifampicin-mutant and its parent strain EILO were analysed. RE-33 is an avirulent isolate used as a modified live vaccine against enteric septicaemia of catfish. Electrophoretic analysis of lipopolysaccharide (LPS) patterns showed high homology between both isolates. Further characterization of LPS by immunoblotting revealed the main differences in LPS composition. The RE-33 isolate lacks the high molecular weight bands of LPS (HMW-LPS). Outer membrane protein analysis also showed some immunological differences between RE-33 and the EILO parent strain. Only two fingerprinting techniques, fatty acid composition analysis and Biolog phenotypic profiles, were able to discriminate between both isolates.