The Effects of Fermentation Process with Acid and Lactic Acid Bacteria Strains on the Biogenic Amine Formation of Wet and Spray-Dried Fish Silages of Discards

The impacts of fermentation process with acid and lactic acid bacteria strains (Lactobacillus plantarum, Lactobacillus brevis, Pediococcus acidilactici, Enterococcus gallinarum, and Streptococcus spp.) on the biogenic amine formation of wet and spray-dried fish silage obtained from whole gibel carp (Carassius gibelio, freshwater discard), whole ponyfish (Equulites klunzingeri, seawater discard), and processing by-products of seabass (Dicentrarchus labrax) were investigated. The results showed that among biogenic amines, cadaverine, putrescine, spermidine, spermine, serotonin, dopamine, and agmatine were predominant in all groups. Significant differences (p < 0.05) in biogenic amine concentrations of wet and spray-dried fish silage were observed. Raw fish and wet silages contained histamine level lower than the allowable limit of 50mg/kg, indicating the use of raw fish material with low microbial counts. In addition, no histamine was detected in spray-dried fish silage, except for seabass by-products with a trace quantity of histamine (<0.04mg/100g). The starter culture used for silage preparation did not effectively retard formation of biogenic amines compared to acid silage. It can be concluded that there is potential use of fermented fish silage as a protein source and possibly as a probiotic ingredient for animal feed in both wet and dry form.     

Synergistic effect of sodium selenite and Pediococcus acidilactici on growth,intestinal bacterial counts,selenium bioavailability,hepatic enzymes and non‐specific immune response in rainbow trout (Oncorhynchus mykiss)

Lactic acid bacteria (LAB) have a crucial role in inorganic selenium metabolism as well as their known desirable effects on fish. In this study, the synergistic effects of dietary sodium selenite and Pediococcus acidilactici on growth performance, intestinal bacterial counts, selenium bioavailability, hepatic antioxidant enzyme thioredoxin reductase activity and hepatic glycolytic enzyme activity that is hexokinase, phosphofructokinase and pyruvate kinase, and non‐specific immune response such as serum lysozyme and complements C₃, C₄ and ACH₅₀ activity in rainbow trout (Oncorhynchus mykiss) were investigated. Thus, a 3 × 3 factorial design experiment was conducted with nine purified diets including three levels of sodium selenite (0, 1 and 2 mg/kg) and three levels of P. acidilactici (0, 7 and 9 log CFU/g). After 8 weeks of feeding, weight gain and specific growth rate were increased by increasing dietary sodium selenite and P. acidilactici levels compared to control (p < .05), whereas feed conversion ratio and condition factor was decreased by increasing dietary sodium selenite and P. acidilactici amounts in comparison with control (p < .05). Survival rate was not significantly affected among the experimental treatments (p > .05). Total cultivable bacterial populations after 4 and 8 weeks of the feeding trial were not significantly different among the dietary treatments, while LAB levels were higher in P. acidilactici‐fed groups than in control and selenium‐fed groups (p < .05). Selenocysteine, methylselenocysteine and selenomethionine levels in the intestine of rainbow trout were increased by increasing the sodium selenite and P. acidilactici levels (p < .05), and selenocysteine was found the most selenium species in the trout intestine. The quantity of total selenium in the whole body, intestine, blood, liver and muscle of rainbow trout were increased by increasing the amounts of sodium selenite and P. acidilactici compared to control (p < .05). Hepatic thioredoxin reductase and hexokinase activity were increased by increasing dietary selenium and P. acidilactici levels in comparison to control (p < .05), whereas phosphofructokinase and pyruvate kinase activity in the liver of rainbow trout were not significantly different between the dietary treatments. Serum lysozyme, complements C₃, C₄ and ACH₅₀ activity were enhanced by increasing dietary selenium and P. acidilactici levels compared to control (p < .05). The most synergistic effects of dietary supplements on growth and metabolism of rainbow trout were obtained at 2 mg/kg sodium selenite and 7 log CFU/g P. acidilactici. The findings revealed the synergistic effect of dietary selenium and P. acidilactici on growth and metabolism in rainbow trout (O. mykiss).     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.     

Effects of dietary supplementation with β‐glucan and synbiotics on growth,haemolymph chemistry,and intestinal microbiota and morphology in the Pacific white shrimp

The goal of this study was to investigate the effects of dietary supplementation with β‐glucan and microencapsulated probiotics (Bacillus subtilis or Pediococcus acidilactici) on growth performance, body composition, haemolymph constituents, and intestinal morphology and microbiota of the Pacific white shrimp Litopenaeus vannamei. Four treatment diets [basal diet (C), β‐glucan‐containing diet (β‐glu), β‐glucan plus B. subtilis‐containing diet (β‐glu+Bs), and β‐glucan plus P. acidilactici‐containing diet (β‐glu+Pa)] were fed to L. vannamei for 90 days. Shrimp fed the β‐glu and β‐glu+Pa diets exhibited similar growth performance and body protein content, which were significantly higher than those of shrimp fed the control diet (P < 0.05). No significant differences in haemolymph triglyceride, cholesterol, protein, haemolymph urea nitrogen or chloride were detected among the experimental diets. However, dietary β‐glucan alone increased the haemolymph glucose level and osmolarity (P < 0.05). Synbiotic supplementation had greater effects on intestinal microbiota and morphology than dietary β‐glucan alone. For example, β‐glu+Bs increased the number of intestinal lactic acid bacteria and decreased the number of Vibrio spp. (P < 0.05), and β‐glu+Pa increased the height of intestinal villi.     

Dietary Houttuynia cordata leaf extract and meal enhances the immunity and expression of immune genes in Labeo rohita (Hamilton, 1822)

A feeding trial was conducted to explore the effect of dietary Houttuynia cordata leaf extract (HCLE) and leaf meal (HCLM) on immunological responses and expression of interferon‐gamma (IFN‐γ) and tumour necrosis factor‐alpha (TNF‐α) gene in Labeo rohita fingerlings. Six isonitrogenous (350 g/kg CP) and isocaloric (17 MJ/kg DE) purified experimental diets were formulated with Houttuynia cordata leaf extract and leaf meal comprising control, C (0 g/kg HCLE and HCLM), E2.5 (2.5 g/kg HCLE), E5 (5 g/kg HCLE), E10 (10 g/kg HCLE), M10 (10 g/kg HCLM) and M20 (20 g/kg HCLM). Labeo rohita fingerlings (3.37 ± 0.23 g) were distributed in six experimental groups in triplicates following the complete random distribution. Fish were fed twice daily with respective experimental diets for a period of 60 days. A significantly (p < .05) lower lactate dehydrogenase, malate dehydrogenase, superoxide dismutase and catalase activities were registered in supplemented groups compared with control group, while respiratory burst and lysozyme activities were significantly (p < .05) higher in E10 group compared with other experimental groups. Haemoglobin, total leucocyte count, total erythrocyte count and haematocrit values were significantly (p < .05) higher in E10 group. The expression of IFN‐γ and TNF‐α in both the kidney and liver was significantly up‐regulated in leaf extract and meal supplemented groups with the highest expression in the fish of E10 group. Overall, these results suggest that the dietary supplementation of ethanolic extract of the Houttuynia cordata leaf at 10 g/kg level can enhance the immune response of L. rohita fingerlings.     

Characterization of spaC‐type Erysipelothrix sp. isolates causing systemic disease in ornamental fish

Since 2012, low‐to‐moderate mortality associated with an Erysipelothrix sp. bacterium has been reported in ornamental fish. Histological findings have included facial cellulitis, necrotizing dermatitis and myositis, and disseminated coelomitis with abundant intralesional Gram‐positive bacterial colonies. Sixteen Erysipelothrix sp. isolates identified phenotypically as E. rhusiopathiae were recovered from diseased cyprinid and characid fish. Similar clinical and histological changes were also observed in zebrafish, Danio rerio, challenged by intracoelomic injection. The Erysipelothrix sp. isolates from ornamental fish were compared phenotypically and genetically to E. rhusiopathiae and E. tonsillarum isolates recovered from aquatic and terrestrial animals from multiple facilities. Results demonstrated that isolates from diseased fish were largely clonal and divergent from E. rhusiopathiae and E. tonsillarum isolates from normal fish skin, marine mammals and terrestrial animals. All ornamental fish isolates were PCR positive for spaC, with marked genetic divergence (<92% similarity at gyrB, <60% similarity by rep‐PCR) between the ornamental fish isolates and other Erysipelothrix spp. isolates. This study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and suggests isolates from diseased ornamental fish may represent a genetically distinct species.     

An evaluation of replacing fish meal with cottonseed meal protein hydrolysate in diet for juvenile blunt snout bream (Megalobrama amblycephala): Growth,antioxidant, innate immunity and disease resistance

This study evaluated effects of fishmeal replacement by cottonseed meal protein hydrolysate (CPH) on growth, antioxidant and immunity of Megalobrama amblycephala and its resistance to Aeromonas hydrophila. Fish (average weight: 38.66 ± 0.08 g) were divided into five groups and fed with five isonitrogenous (320 g/kg crude protein), isolipidic (70 g/kg crude fat) and isocaloric (17.8 MJ/kg gross energy) diets replacing fishmeal with 0%, 1%, 3%, 5% and 7% CPH, respectively. The control diet contained 60 g/kg fishmeal. Final weight and weight gain of fish fed 5% and 7% CPH were significantly lower than that of fish fed control diet (p < 0.05). 3% CPH significantly increased total protein, globulin, acid phosphatase, C3 and C4 contents of plasma and total‐superoxide dismutase and catalase activities of liver (p < 0.05), as well as upregulated the antimicrobial peptides 1 and 2 (Leap‐1 and Leap‐2) expression levels in liver and spleen (p < 0.05). After challenge, the lowest mortality was observed in fish fed 3% CPH, and it was significantly lower than that in fish fed the 7% CPH (p < 0.05). In conclusion, dietary fishmeal replacement by 3% CPH could increase antioxidative capacity, as well as enhance immunity of fish.     

Effects of dietary vitamin C and vitamin E on the growth,antioxidant defence and digestive enzyme activities of juvenile discus fish (Symphysodon haraldi)

A 56‐day experiment was carried out to investigate the effects of dietary vitamin C and vitamin E on the growth, antioxidant status and digestive enzyme activities of discus fish (Symphysodon haraldi; initial body weight: 7.96 ± 0.61 g and body length: 5.45 ± 0.65 cm). Animals were fed with 13 different diets including one control diet and 12 treatment diets containing four levels of vitamin C (magnesium‐L‐ascorbyl‐2‐phosphate; 40, 80, 120 and 160 mg/kg) crossed with three levels of vitamin E (DL‐α‐tocopheryl acetate; 40, 80 and 120 mg/kg). The results showed that the fish fed diets containing additional vitamin C (40 mg/kg) and vitamin E (80 mg/kg) showed higher specific growth rate, length growth rate, total antioxidant capacity and protease activity but had lower feed conversion ratio and total superoxide dismutase activity than those fed the control diet. Collectively, these findings suggest that the inclusion of additional 40 mg/kg of vitamin C and 80 mg/kg of vitamin E in the basal diet could have beneficial effect on the growth, antioxidant defence and digestion of S. haraldi.     

Effects of dietary coated protease on growth performance,feed utilization,nutrient apparent digestibility,intestinal and hepatopancreas structure in juvenile Gibel carp (Carassius auratus gibelio)

This study was conducted to investigate the effects of dietary protease on growth performance, feed utilization, whole‐body proximate composition, nutrient digestibility, intestinal and hepatopancreas structure of juvenile Gibel carp, Carassius auratus gibelio (mean weight 8.08 ± 0.18 g). Six diets were prepared, including a positive control diet (dietary protein 350 g/kg, PC), one negative control diet (dietary protein 33 g/kg, NC) and four protease supplementations diets, which were 75, 150, 300 and 600 mg/kg protease NC diet. After 12 weeks of diet feeding in indoor recycle aquarium tanks, no significant difference (p > .05) was found on growth performance between fish fed diet with 75–600 mg/kg protease and the PC group. Compared with the fish fed the NC diet, the specific growth rate of fish fed 300 mg/kg protease increased significantly (p < .05), as well as protein efficiency ratios (p < .05), while feed conversion was the opposite (p < .05). The nutrient digestibility of crude protein and lipid was higher (p < .05) in fish fed 150 mg/kg protease diet than the PC diet. Whole‐body proximate composition of fish was not affected (p > .05) by the dietary treatment. Serum alkaline phosphatase and albumin were significantly affected by dietary protease (p < .05), while the content of total protein, glucose, triglyceride, total cholesterol, aspartate aminotransferase and alanine aminotransferase activities in serum was not affected (p > .05). Foregut muscular thickness was thinner (p < .05), when the fish fed diets supplementation of protease in 150 or 600 mg/kg diet than the NC diet. Protease activities in hepatopancreas and foregut were higher (p < .05), in the fish fed 150 or 300 mg/kg protease diet than the fish fed the PC diet, but those in the mid‐ and hindgut were not significantly affected (p > .05) by the dietary treatments. Based on the regression analysis of weight gain rate, the optimal dietary inclusion level of protease was 400 mg/kg in the diet for juvenile Carassius auratus gibelio.     

Augmenting fish health using Emblica officinalis against triarylmethane dye induced blood toxicity in Cyprinus carpio

Blood is a sensitive bioindicator of stress observed by organisms in response to toxicants. Erythrocytic morphological abnormalities serve as a reliable determinant of fish health. Malachite green, a multifunctional dye was assessed for toxicity in Cyprinus carpio. Dye‐induced blood toxicity has been determined through biochemical and ultra structural endpoints at 15, 30 and 60 days in the fish. Further, an attempt has been made to combat the adverse effects of the toxicant through supplementation with Emblica officinalis. Electron microscopic study depicted erythrocytic remodelling in form of surface blebbing, lobopodial projections and shrinkage; appearance of crenate, acanthocytes, echinocytes, dacrocytes, spherocytes and rhomboidal cells. Hepatic marker enzymes showed significantly (p < 0.05) elevated levels of biomarker enzymes, alkaline phosphatase, serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase. Contrarily feeding with E. officinalis extract (1,000 mg/kg feed) significantly attenuated biochemical and morphological alterations, indicating the protective efficacy of E. officinalis. Both toxicity and attenuation observed were in exposure‐dependent manner.     

Efficacy of single and multiple doses of fenbendazole against gill parasites (Dactylogyrus sp.) of Labeo rohita (Hamilton, 1822) and its physio‐metabolic effects on the fish

The study was conducted to investigate the efficacy and physio‐metabolic responses of single and multiple doses of fenbendazole (FBZ) in Labeo rohita against gill parasite (Dacylogyrus sp.). Two Isoenergetic (395 Kcal DE/100 g) and isonitrogenous (35% CP) purified diets viz. control (without FBZ) and treatment (0.2% FBZ) were prepared. The efficacy of FBZ was measured after administration of single dose (20 mg/kg b. wt.) on 0th day and multiple doses of FBZ, that is, 20 mg/kg b. wt. on 0th, 3rd and 7th day through diet. Although oral administration of FBZ at single dose could significantly reduce the parasite in fish in comparison with the control group, 100% efficacy of the drug was not achieved up to 15th day of treatment. On the other hand, multiple doses of FBZ could show almost 100% efficacy on 15th day in terms of elimination of parasites from gill. The drug induced oxidative stress in examined fish. There was no significant change in activities of hepatic and muscle aspartate aminotransferase and alanine aminotransferase of L. rohita up to 15th day after oral administration of FBZ at single dose. But both single and multiple doses of FBZ could significantly enhance the hepatic superoxide dismutase, catalase and glutathione‐S‐transferase activities in fish up to 7th day of drug administration and significantly decreased thereafter up to 15th and/or 30th day. It is concluded that oral administration of FBZ at multiple doses (20 mg/kg b. wt.) on 1st, 3rd and 7th day could be effective against Dactylogyrus sp. infection in L. rohita.     

Effects of dietary probiotic Lactobacillus plantarum and whey protein concentrate on the productive parameters,immunity response and susceptibility of Nile tilapia,Oreochromis niloticus (L.), to Aeromonas sobria infection

Immunostimulatory feed supplements have an increasingly interest in aquaculture management. Generally, an individual supplement was used in fish diets but it is expected that the use of multi‐supplements may show synergistic enhancements in fish performance, health, and immunity. Therefore, the present investigation was carried out to evaluate the use of dietary probiotic Lactobacillus plantarum and whey protein concentrate (WPC) in practical diets for Nile tilapia, Oreochromis niloticus. Hence, probiotic L. plantarum, WPC and their mixture were incorporated into a basal fish diet (300 g/kg crude protein) as follows: T1 = a basal control diet, T2 = a basal diet containing L. plantarum, T3 = a basal diet containing 1.0 g WCP/kg diet and T4, T5 or T6 = basal diets containing probiotic L. plantarum + 1.0, 2.0 or 3.0 g WCP/kg diet, respectively. Fish (15.2 ± 0.6 g) were fed on one of the tested diets up to apparent satiation twice a day for 60 days. After that, fish were intraperitoneally injected with pathogenic bacteria Aeromonas sobria and fish mortality was observed for 10 days postchallenge. Fish growth and feed intake were significantly improved by dietary probiotic L. plantarum (T2) and/or WPC (T3) over the control group (T1), and highest fish performance was observed in T5–T6 fish groups. Similarly, highest values of haematocrit, glucose, total proteins, albumin, and globulin were significantly observed in T5–T6 fish groups. Likewise, fish fed dietary probiotic L. plantarum (T2), WPC (T3), and their mixture (T4–T6) showed antioxidants and immune‐stimulating activities better than the control group. Fish fed the control diet were more susceptible to A. sobria infection showing highest fish mortality (75.0%). Meanwhile, dietary probiotic L. plantarum (T2), WPC (T3), and their mixture (T4–T6) enhanced significantly the fish resistance to A. sobria infection resulting in maximum values of relative percent of fish survival (73.3%–80.0%) in T5–T6 groups. The present investigation recommended the use of probiotic L. plantarum with 2.0 g WPC/kg diet to improve the growth, antioxidant, immunity responses and tolerance of Nile tilapia to A. sobria infection.     

Potential of processed animal protein versus soybean meal to replace fish meal in practical diets for European catfish (Silurus glanis): growth response and liver gene expression

A 9‐week study was conducted to evaluate the potential of processed animal protein (PAP) in comparison with soybean meal (SM) to replace fish meal (FM) in practical diets for European catfish, Silurus glanis, on growth performance, liver transaminase activities and expression of growth‐related genes. Seven isonitrogenous (440 g kg^‐1 crude protein) and isoenergetic (16.70 MJ/kg) practical diets were formulated by replacing 0 (control), 30%, 60% and 100% FM with either SM or PAP. Each diet was randomly assigned to triplicate groups of 25 fish per cages fixed in the pond. Fish fed the diet substituted 100% FM from SM or PAP had lower (p < .05) growth performance, feed efficiency and protein retention compared to control and other groups. The decreased growth performance occurred concomitant with a decline in the expression level of liver GHR/IGF‐I axis genes. The gene expression and activity of liver protein metabolism enzymes also positively correlated with growth performance. Findings of this study indicated that both SM and PAP were equally effective in replacing FM up to 60% in the practical diet of Silurus glanis. Total replacement of FM either with SM or PAP induced negative influences on growth and feed utilization.     

Evaluation of dried fermented biomass as a feed ingredient in plant‐based practical diets for juvenile Pacific white shrimp Litopenaeus vannamei

Two growth trials were designed to evaluate the utilization of dried fermented biomass (DFB) in commercial type feed formulation for Pacific white shrimp, Litopenaeus vannamei. In trial 1, four experimental diets were formulated to utilize increasing levels (0, 25, 50 and 100 g/kg) of spray‐dried fermented biomass (SDFB) as a replacement of fish meal (FM). Results indicated that SDFB can be utilized up to 50 g/kg as a substitution for FM without causing growth depression in shrimp. However, dietary SDFB supplementation at 100 g/kg significantly reduced the weight gain (WG) of shrimp and increased feed conversion ratio (FCR). This reduction in performance is likely due to palatability or nutrient imbalances of the feed. In trial 2, nine experimental diets were formulated with increasing levels (0, 20, 40, 60 and 120 g/kg) of spray‐dried (S) or granular (G) DFB to replace soy protein concentrate (SPC) or SPC + corn protein concentrate (CPC). This allowed the comparison between spray‐dried and ring‐dried products. Ring drying produced a granular product, reducing dust and increasing product particle size. Shrimp fed with diet containing 20 g/kg GDFB performed the best in terms of final mean weight, WG and FCR. Significantly reduced growth and increased FCR were observed in shrimp fed diets containing 60 and 120 g/kg SDFB. Lipid content of whole body was significantly reduced when GDFB was incorporated at 120 g/kg. No significant differences were detected in survival, protein retention efficiency as well as protein and ash contents of the whole shrimp. Results from analysis of covariance indicated that the processing method (covariant) had a significant effect on final mean weight, WG and FCR. In general, shrimp fed with diet containing granular product performed better as compared to those fed with diets utilizing spray‐dried product. GDFB can be utilized in the diets up to 120 g/kg in practical shrimp feeds as a substitute for SPC and CPC without compromising the growth of shrimp. However, a significant reduction in WG was observed in the diets containing 60 and 120 g/kg SDFB. The results in the current study demonstrate that processing changes to produce a granular product produced an improved feed ingredient for shrimp.     

Mycobacteria in aquarium fish: results of a 3‐year survey indicate caution required in handling pet‐shop fish

Fish are commonly infected with non‐tuberculous mycobacteria (NTM), which should be regarded as potential pathogens when handling aquarium fish and equipment. This study examined 107 aquarium fish from pet shops. Cultivation of the fish samples using different selective media was conducted for identification of NTM. Isolates were identified using the GenoType Mycobacterium common mycobacteria and additional species assays, sequencing of the 16S rRNA and rpoB genes, and real‐time PCR assay for identification of Mycobacterium (M.) marinum. Among the investigated fish, 79.4% (85/107) were positive for mycobacteria, with 8.2% (7 of 85) having two mycobacterial species present. Among the positive fish, the common pathogens M. marinum, Mycobacterium fortuitum (M. fortuitum group) and Mycobacterium chelonae were identified in approx. 90% of fish and other NTM species in 10%, including Mycobacterium peregrinum/septicum, Mycobacterium gordonae, Mycobacterium arupense, Mycobacterium kansasii, Mycobacterium ulcerans and Mycobacterium setense. The well‐known human pathogen M. marinum was present in 10.6% of the positive fish (9 of 85). The species of mycobacteria identified in the study are not only recognized as aquarium fish pathogens, but can also cause pathology in humans. Microbiological and clinical communities should therefore be sensitized to the role of NTM in infections associated with exposure to aquarium fish.     

Comparative genomic insights into the taxonomy of Edwardsiella tarda isolated from different hosts: Marine,freshwater and migratory fish

Edwardsiella tarda, a Gram‐negative member of the family Enterobacteriaceae, has been isolated from many animal species worldwide, especially fish species. Its broad host range indicates the diversity in taxonomy, which attracted the attention of many researchers. Here, we added genome of E. tarda strain isolated from freshwater fish to comparative genomics study for the first time. We sequenced and assembled the genome of E. tarda ASE201307 which was isolated from freshwater Asian swamp eel. ASE201307 genome contained a single circular chromosome of 3.68M with G+C 57.09% content. Comparative genomics including SNP calling, synteny block, Core/Pan genes analysis and phylogeny analysis was conducted among ASE201307 and other Edwardsiella strains isolated from different fish species. Results of SNP analysis and synteny block demonstrated the close relative of ASE201307, FL95.01 and DT which were all isolated from freshwater fish. In further analysis heat map of dispensable genes and phylogenetic tree, all E. tarda strains were divided into two groups. One was isolated from freshwater fish and the other was isolated from marine/migratory fish. Based on all studies above, we proposed the living environment of hosts as a new taxonomic character and divided E. tarda isolated from diseased fish into freshwater group and marine/migratory group.     

Nutritional evaluation of distiller's dried grain with soluble as replacement to soybean meal in diets of milkfish,Chanos chanos and its effect on fish performance and intestinal morphology

A 90‐day feeding trial was conducted on milkfish, Chanos chanos with an initial mean body weight of 3.07 ± 0.17 g (mean ± standard error of mean). Six treatment diets were formulated to contain 0 g/kg (Diet 1), 150 g/kg (Diet 2), 25 g/kg (Diet 3), 300 g/kg (Diet 4), 350 g/kg (Diet 5) and 450 g/kg (Diet 6) distiller's dried grain with soluble (DDGS). All the dietary treatments were isonitrogenous (350 g/kg crude protein) and isolipidic (6% crude lipid). Result of the feeding trial indicated that growth rates, feed intake and feed efficiency were not significantly (p > .05) affected by inclusion levels of DDGS by up to 450 g/kg in the feed. Proximate body composition (crude protein, crude lipid, ash, fibre) in fish fed the dietary treatments were not significantly (p > .05) affected as well. The DDGS when used as a milkfish ingredient has a protein digestibility of 910 g/kg, fat disgetsibility of 850 g/kg, carbohydrate digestibility of 750 g/kg and a dry matter digestibility of 520 g/kg Results from the intestinal morphology displayed no apparent pathological changes in the digestive tract of fish fed all dietary treatments. These results indicate that DDGS can be efficiently utilized by milkfish by up to 450 g/kg without negatively affecting performance parameters and intestinal morphology.     

Identification and pathogenicity study of emerging fish pathogens Acinetobacter junii and Acinetobacter pittii recovered from a disease outbreak in Labeo catla (Hamilton, 1822) and Hypophthalmichthys molitrix (Valenciennes, 1844) of freshwater wetland in West Bengal,India

The disease outbreaks in aquaculture system of wetlands are the major cause of fish mortality. Among various bacterial septicaemic diseases, fish mortality caused by Acinetobacter spp. is recently reported in different fish species. Fish disease outbreak was investigated in a wetland of West Bengal, India to identify the aetiological factors involved. The moribund fish were examined and subjected to bacterial isolation. Two bacterial causative agents were identified as Acinetobacter junii and Acinetobacter pittii by biochemical characterization and 16S rRNA gene amplification. Both the isolates were oxidase‐negative, nitrate‐negative, catalase‐positive and indole‐negative. The molecular identification using 16S rRNA gene sequencing and phylogenetic tree analysis further confirmed the two Acinetobacter spp. with 97%–99% similarity. The antibiotic resistance patterns of these two bacteria revealed that both of them were resistant to β‐lactam, cefalexin, cephalothin, amoxyclav, cefuroxime, cefadroxil, clindamycin, vancomycin and penicillin. In addition, A. pittii was also resistant to other antibiotics of cephams group such as ceftazidime and cefotaxime. In the challenge experiment, both A. junii and A. pittii were found to be pathogenic with LD₅₀ of 1.24 × 10⁵ and 1.88 × 10⁷ cfu/fish respectively. Histopathological examination of gill, liver and kidney revealed prominent changes supporting bacterial septicaemia. The investigation reports for the first time on concurrent infection by A. junii and multidrug‐resistant (MDR)‐A. pittii as emerging fish pathogens to cause severe mortality in Labeo catla and Hypophthalmichthys molitrix in a freshwater wetland.     

Growth and economic analysis of freshwater prawn,Macrobrachium rosenbergii (de Man), produced with feeds substituting sunflower cake for fish meal,soya bean meal and mustard oil cake

A feeding trial with Macrobrachium rosenbergii was carried out to test the effectiveness of sunflower cake (SFC) as a replacement for conventional protein sources. Four isonitrogenous (~30% crude protein) and isocaloric (388–402 kcal) diets were formulated. Diet‐1 was a control made with conventional ingredients including fish meal, soya bean meal and mustard oil cake. Diet‐2, Diet‐3 and Diet‐4 were made with 30%, 40% and 50% SFC, respectively, replacing the usual protein sources used in Diet‐1. After 100 days, diets with SFC inclusion showed favourable growth and economic performance trends compared to control diet‐1, but without significant differences (p > .05). Better production (572.89 ± 62.99 kg/ha) was observed with Diet‐4 followed by Diet‐2, Diet‐3 and Diet‐1. Diet‐4 revealed the best feed conversion ratio (2.56 ± 0.24), specific growth rate (1.18 ± 0.05) and protein efficiency ratio (1.14 ± 0.10) among these diets. Whole carcass composition analysis revealed highest crude protein (18.19%) and lipid (4.43%) content in prawns fed Diet‐3 followed by Diet‐4. Highest total income, gross margin and benefit cost ratio (BCR) per hectare from prawns were US$5,759, US$2,623 and 1.84, respectively, found in Diet‐4. This study demonstrates that 30%–50% inclusion of SFC has favourable impacts on productivity and profitability compared with controls, with 50% SFC inclusion as the most cost‐effective diet for prawns without apparent compromise in growth, flesh quality or economics. The performances of the diets suggest that SFC may be a suitable substitute for fishmeal, soya bean meal and mustard oil cake in prawn feed. The effectiveness of higher SFC concentrations should also be investigated.