Methods of microbial control in marine fish larval rearing: clay‐based turbidity and passive larval transfer

This study focused on methods to reduce bacterial loads in the larval culture tanks of California yellowtail (Seriola lalandi). We conducted two trials to evaluate methods to minimize bacterial loads in the larval rearing water. The first trial examined the use of bentonite clay as a turbidity agent to replace algae in a green water‐type environment. This trial consisted of three treatments: (1) clay with continuous feeding (CCO), (2) clay with batch feedings (CBA) and (3) algae paste with batch feedings (ALG). The results showed that both clay treatments had significantly fewer Vibrio colonies in the water column (CBA – 180 ± 78; CCO – 377 ± 120 CFU mL^?1) than the ALG treatment (5692 ± 2396 CFU mL^?1) after 14 days of culture. Survival was significantly higher in the CCO treatment (14.1 ± 2.6%) than either the CBA (2.3 ± 0.5%) or ALG treatments (2.8 ± 1.5%). The second trial attempted to limit bacterial loading in the larval culture tank by passively transferring the larvae into an adjacent, clean tank at 1, 5 and 9 days post hatch during the first 2 weeks of culture. The results from this trial showed that after 12 days of culture, water in the transfertank had fewer Vibrio colonies (1025 ± 541 CFU mL^?1) than the water in the control tanks (1962 ± 1415 CFU mL^?1). Also, survival was significantly higher among larvae that were transferred (43.9 ± 13.5%) than in the control tanks (23.1 ± 6.3%).     

Development and validation of a real‐time PCR assay for the detection of Aeromonas salmonicida

A real‐time PCR assay using a molecular beacon was developed and validated to detect the vapA (surface array protein) gene in the fish pathogen, Aeromonas salmonicida. The assay had 100% analytical specificity and analytical sensitivities of 5 ± 0 fg (DNA), 2.2 × 10⁴ ± 1 × 10⁴ CFU g^?1 (without enrichment) and 40 ± 10 CFU g^?1 (with enrichment) in kidney tissue. The assay was highly repeatable and proved to be robust following equivalency testing using a different real‐time PCR platform. Following analytical validation, diagnostic specificity was determined using New Zealand farmed Chinook salmon, Oncorhynchus tshawytscha (Walbaum), (n = 750) and pink shubunkin, Carassius auratus (L.) (n = 157). The real‐time PCR was run in parallel with culture and all fish tested were found to be negative by both methods for A. salmonicida, resulting in 100% diagnostic specificity (95% confidence interval). The molecular beacon real‐time PCR system is specific, sensitive and a reproducible method for the detection of A. salmonicida. It can be used for diagnostic testing, health certification and active surveillance programmes.     

Anaesthetic efficacy of MS‐222 and AQUI‐S® in advanced size fry of rohu,Labeo rohita, (Hamilton‐Buchanan)

Anaesthetic agents are very useful for reducing the stress caused by handling, sorting, transportation, artificial reproduction, tagging, administration of vaccines and surgical procedures in fish. The efficacy of two anaesthetics: MS‐222 and AQUI‐S^® were tested on rohu, Labeo rohita advanced size fry. The lowest effective doses that produced induction in 3 min or less and recovery times 5 min or less and meet the most criteria of good anaesthetic characteristics were 125 mg L^?1 of MS‐222, and 30 mg L^?1 of AQUI‐S^® in rohu, Labeo rohita advanced size fry. Induction times were significantly decreased with increased in the concentrations of any of the two tested anaesthetic agents. The lowest doses suitable for transportation of rohu advanced size fry observed were: 10–15 mg L^?1 of MS‐222 and 2.5 mg L^?1 AQUI‐S^®. Both anaesthetics showed promising to be used as anaesthetics for handling and transportation in rohu (Labeo rohita) advanced fry.     

Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Immune responses and gut morphology of Senegalese sole (Solea senegalensis,Kaup 1858) fed monospecies and multispecies probiotics

The current study aimed to determine the effects of dietary probiotic supplementation on growth, gut morphology and non‐specific immune parameters in Senegalese sole (Solea senegalensis) juveniles during a 1‐month trial. Fish were fed for 1‐month two diets with 1.0 or 4.6 × 10⁶ CFU kg^?1) of probiotic A (Bacillus sp., Pediococcus sp., Enterococcus sp. and Lactobacillus sp.) and two diets with 3.5 or 8.6 × 10⁵ CFU kg^?1 of probiotic B (Pediococcus acidilactici) and tested against an unsupplemented diet (control). Growth performance, as well as respiratory burst activity, nitric oxide (NO), alternative complement pathway (ACH50), lysozyme and peroxidase activities, was not affected by the dietary treatments. Probiotic supplementation tended to increased growth homogeneity between tanks having diet A₁ the best possible alternative to decrease costs associated to size grading. Villous length and number of goblet cells of the anterior intestine did not vary among treatments. Muscle duodenal layer was significantly thicker in fish fed probiotic A compared to probiotic B, when included at the lowest level (A₂ versus B₂). The current study indicate that the use of the multispecies probiotic at 1.0 × 10⁶ CFU kg^?1 might enhance protection against pathogen outbreak and increase nutrient absorption, whereas at the highest concentration could reduced size dispersion among tanks.     

Synbiotic effect of Bacillus mycoides and organic selenium on immunity and growth of marron,Cherax cainii (Austin, 2002)

The present feeding trial examined the effect of synbiotic use of Bacillus mycoides and organic selenium (OS) as Sel‐Plex on marron immunity, growth and survival. The marron were cultured in recirculated tanks and fed test diets consisting of a basal diet; basal diet supplemented with B. mycoides (10⁸ CFU g^?1 of feed); basal diet supplemented with OS (Sel‐Plex) (0.2 g kg ^{? 1} of feed) and basal diet supplemented with synbiotic (B. mycoides at 10⁸ CFU g^?1 and OS 0.2 g kg ^{? 1} feed) diet, in triplicate. The effect of the prebiotic OS (Sel‐Plex) on the growth rate of B. mycoides was also studied in vitro. The results suggested that synbiotic use of B. mycoides and OS significantly improved some immune parameters of marron, particularly the glutathione peroxidase, and to some extent total haemocyte counts. However, the synbiotic feed did not synergistically improve marron growth, in fact the use of B. mycoides‐supplemented diet alone demonstrated significantly higher growth in marron compared with the growth of marron fed on other test diets. Supplementation of the basal diet with host origin B. mycoides significantly increased the intestinal bacterial population (3.399 ± 825 CFU g ^{? 1} of gut) in marron compared with other diets. Organic selenium as Sel‐Plex in Trypticase Soya Broth also confirmed that OS did not increase the amount of growth of B. mycoides and resulted in a lower intestinal bacterial population in the synbiotic diet‐fed marron. In conclusion, synbiotic of OS and B. mycoides may improve a particular immune parameters of marron and to a lesser extent their growth.     

Stocking density effects on growth and production of the threatened silurid catfish,Mystus cavasius (Hamilton) fingerlings in nursery ponds

The fingerling‐rearing experiment of the threatened catfish, Mystus cavasius was carried out at different stocking densities in earthen nursery ponds. Twelve‐day‐old fry were stocked at 200 000 ha^?1 in treatment‐1 (T₁), 250 000 ha^?1 in treatment‐2 (T₂) and 300 000 ha^?1 in treatment‐3 (T₃) respectively. The mean length and weight of fry at stocking was 1.24 ± 0.25 cm and 0.11 ± 0.04 g respectively. Fry in all the experimental ponds were supplemented with SABINCO nursery feed for the first 14 days and starter‐I feed for days 15–56. The physico‐chemical parameters and plankton population of pond water were within the suitable level for fish culture. Growth in terms of final weight, final length, weight gain, length gain and specific growth rate and survival of fingerlings were significantly higher in T₁ than those in T₂ and T₃. Feed conversion rate was significantly lower in T₁ followed by T₂ and T₃ in that order. Significantly higher number of fingerlings was produced in T₃ than that in T₂ and T₁. Even then, consistently higher net benefits were obtained from T₁ than those from T₃ and T₂. Among the treatments evaluated, 200 000 fry ha^?1 was the best stocking density considering the highest growth, production and net benefits of fingerlings of M. cavasius in nursery ponds.     

Efficacy of garlic (Allium sativum) extract applied as a therapeutic immersion treatment for Neobenedenia sp. management in aquaculture

Garlic, Allium sativum L., extract administered as a therapeutic bath was shown to have antiparasitic properties towards Neobenedenia sp. (MacCallum) (Platyhelminthes: Monogenea) infecting farmed barramundi, Lates calcarifer (Bloch). The effect of garlic extract (active component allicin) immersion on Neobenedenia sp. egg development, hatching success, oncomiracidia (larvae) longevity, infection success and juvenile Neobenedenia survival was examined and compared with freshwater and formalin immersion. Garlic extract was found to significantly impede hatching success (5% ± 5%) and oncomiracidia longevity (<2 h) at allicin concentrations of 15.2 μL L^?1, while eggs in the seawater control had >95% hatching success and mean oncomiracidia longevity of 37 ± 3 h. At much lower allicin concentrations (0.76 and 1.52 μL L^?1), garlic extract also significantly reduced Neobenedenia infection success of L. calcarifer to 25% ± 4% and 11% ± 4%, respectively, compared with 55% ± 7% in the seawater control. Juvenile Neobenedenia attached to host fish proved to be highly resistant to allicin with 96% surviving 1‐h immersion in 10 mL L^?1 (15.2 μL L^?1 allicin) of garlic extract. Allicin‐containing garlic extracts show potential for development as a therapy to manage monogenean infections in intensive aquaculture with the greatest impact at the egg and larval stages.     

Preliminary study on broodstock rearing,induced breeding and grow‐out culture of the sea cucumber Holothuria scabra in Sri Lanka

Experiments were conducted to identify suitable methods for broodstock rearing, induced breeding and grow‐out culture of Holothuria scabra in Sri Lanka. Two hundred and seventy‐two brooders (500–600 g) collected from off Mannar were individually packed in oxygen‐filled polythene bags with and without sea water and transported to a sea cucumber hatchery at Kalpitiya. Lagoon pens, sand‐filled fibreglass tanks and bare tanks were used in triplicates to maintain brooders. Spawning was initiated using air dry, water jet and thermal‐stimulation methods. Hatchery produced juveniles with an average weight of 11 ± 5 g were reared (2 individuals m^?2) in lagoon pens, mud ponds and fibreglass tanks in triplicates. The significantly high evisceration rate was observed when brooders were transported without sea water (t‐test, P < 0.05). Brooders maintained in bare tanks showed a significant weight reduction than the brooders in sand‐filled tanks and lagoon pens (anova , P < 0.05, d.f. = 2). Thermal stimulation (ambient temperature ± 3–5°C) was found to be the most successful method of spawning initiation of H. scabra. The mean (±SD) percentage males and females participated for spawning per trial was 9.2 (±10) and 4.6 (±5.6) respectively. On an average, 1.16 millions of eggs (±1.03 SD, n = 5) were obtained per spawning trial. H. scabra juveniles reared in tanks showed significantly lower growth rate than the juveniles in pens and ponds (anova , P < 0.05). Lagoon pens and sand‐filled tanks are suitable to maintain brooders and lagoon pens can be successfully used for mass rearing of juveniles.     

High density rearing of rohu,Labeo rohita (Hamilton) from spawn to fry in concrete tanks: effect of daily feed ration and soil base on fry growth and survival

Rearing of rohu spawn to fry size was carried out at high density (2000 spawn m^?2) in large concrete nursery tanks (10 m × 5 m × 1.2 m) to standardize the daily ration of supplementary feed and to assess the impact of soil base on the fry performance. In Set‐1, supplementary feed was provided at 100% (control), 75% and 50% of the daily ration that is commonly used in earthen carp nursery ponds and named as the three treatments T‐1, T‐2 and T‐3 respectively. Although in Set‐II, performance of fry were evaluated in tanks provided with and without soil base. The fry survival was to the tune of 45–59% at this high density of 2000 spawn m^?2, indicating the efficacy of the concrete nursery system for high density seed rearing. In Set‐I, T‐2 receiving 75% of the control ration exhibited similar fry growth as that of T‐1, but its fry survival (58.8%) was 10% higher than the latter. This indicated that supplementary feed in a concrete nursery can be reduced up to 25% of that provided in earthen nursery system while ensuring a higher fry yield. However, further reduction up to 50% of the control ration in T‐3 yielded lower fry survival and growth (P < 0.05) revealing inadequacy of the available feed. In Set‐II, treatment without soil base (T‐ns) yielded similar fry growth but 10% higher survival compared to that with the soil base (T‐s) revealing non‐essentiality of soil base in concrete tanks for such short‐term nursery rearing of rohu.     

Effect of dietary supplementation of periphyton on growth,immune response and metabolic enzyme activities in Penaeus monodon

A 60‐day indoor trial was conducted to study the effect of periphyton supplementation on metabolic and immune responses in tiger shrimp, Penaeus monodon. Periphyton developed over bamboo substrate in outdoor tanks (15 m²) was used as dietary supplement for P. monodon (2.02 ± 0.04 g) reared in 1000 L FRP tanks. Graded levels of periphyton were included in shrimp basal diets: 0% (P0), 3% (P3), 6% (P6), 9% (P9) and P0 diet with natural periphyton (NP) over bamboo substrate. At the end of the trial, P6 and NP showed significantly higher (P < 0.01) growth rate, 23.9% and 20%, respectively, compared with control, P0. Comparatively, lower level of metabolic enzymes, such as lactate dehydrogenase, malate dehydrogenase, aspartate aminotransferase and alanine aminotransferase, was recorded in treatments P3, P6 and NP compared with control, P0. The periphyton‐supplemented group, P3 had significantly higher (P < 0.05) superoxide dismutase (15.83 ± 0.96) and catalase activity (15.73 ± 0.69) compared to 6.88 ± 2.84 and 9.15 ± 0.67 unit mg^?1 protein min^?1_, respectively, in P0. Similarly, higher total haemocyte counts, 32.58 ± 1.30, 28.51 ± 3.12 and 27.26 ± 4.43 × 10⁶ cells mL^?1_, were recorded in P6, NP and P3, respectively, compared to P0, 23.57 ± 1.80 × 10⁶ cells mL^?1. After challenge with Vibrio harveyi, P3 recorded the highest relative percentage survival 67% followed by NP (58%) and P6 (42%) compared with control. However, treatment with highest periphyton inclusion (P9) did not differ significantly with P0 on growth and immunological parameters. This study indicates that periphyton supplementation at 3–6% level improves growth, immune response and metabolic activities in P. monodon.     

Pond production of Pacific white shrimp,Litopenaeus vannamei,fed grain distillers dried yeast

The objective of this study was to demonstrate the feasibility of four diets formulated to contain increasing levels (0, 50, 100 and 150 g kg^?1 of diet) of grain distillers dried yeast (GDDY) in production diets for Litopenaeus vannamei, reared in outdoor tanks or production ponds. The production pond trial was carried out in 16, 0.1‐ha ponds using four replicates per diet. Juvenile shrimp (38.1 ± 4.26 mg, initial weight) were stocked at 30 shrimp m^?2 for a 16‐week period. The same four diets and a commercial reference diet were offered to shrimp maintained in outdoor tanks over a 12‐week period. A total of 20 tanks were stocked with juvenile shrimp (3.05 ± 0.22 g, initial weight) obtained from production ponds at a density of 30 shrimp per tank (40 shrimp m^?2). At the conclusion of these trials, mean final weight ranged from 19.77 to 23.05 g, yield ranged between 4760 and 5606 kg ha^?1, survival ranged from 69.6% to 89.4%, and feed conversion ratio (FCR) was between 1.02 and 1.23. Shrimp reared in the outdoor tanks confirmed the results of the pond trial. Mean final weight ranged between 18.12 and 18.97 g, survival ranged from 93.3% to 98.3%, and FCR was between 1.25 and 1.29. In both trials, there were no significant differences regarding mean final weight, FCR and survival among dietary treatments. Based on this study, GDDY up to 150 g kg^?1 of diet can be used in L. vannamei commercial feed formulation.     

Commercial Bacillus probiotic supplementation of rainbow trout (Oncorhynchys mykiss) and brown trout (Salmo trutta): growth,immune responses and intestinal morphology

Probiotic administration is associated with the enhancement of host resistance to environmental and nutritional stressors, improving survival and growth rates. This study was carried out to evaluate the effects of dietary supplementation of Bacillus subtilis and Bacillus cereus toyoi on growth performance, innate immune responses and gut morphology of two trout species feeding a commercial diet high in soybean meal. A commercial probiotic (4.2 × 10⁹ CFU g^?1 of additive) was supplemented to the experimental diets at 0% (control), 0.03% (P₁; 6 × 10³ CFU g^?1 of diet) or 0.06% (P₂; 1.5 × 10⁶ CFU g^?1 of diet) and fed to brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) for 9 and 20 weeks respectively. Rainbow trout showed significantly better growth performance than brown trout, regardless of the dietary treatment. No effect of dietary probiotic supplementation was detected on growth performance, body composition or innate immune parameters (plasma lysozyme, alternative complement and peroxidase activities). In both species, after 9 weeks, intestinal lamina propria and submucosa were widened, with increased presence of inflammatory cells, significantly higher in groups fed probiotics. This inflammatory process, with villi and enterocytes noticeably damaged compared with the control group, was more pronounced in brown trout. Under the current trial conditions, the B. subtilis + B. cereus toyoi had no positive impact in either trout species, on the contrary a harmful effect was observed.     

The effect of feeding rate on the growth performance of green sturgeon (Acipenser medirostris) fry

Four one‐week growth trials were conducted on green sturgeon fry to determine the effect of feeding rate on their growth performance at 18 °C when they were fed a salmonid soft moist feeds containing 445–457 g kg^?1 of crude protein and 201–207 g kg^?1 of lipid. The fry used in Trials I‐IV were 5–8 weeks after their initiation of exogenous feeding. Their average initial body weights were 1.63 ± 0.01, 2.63 ± 0.03, 5.08 ± 0.08 and 7.49 ± 0.05 g, respectively. Six feeding rates used were as follows: 2.5–15.0% body weight per day (% BW day^?1) with a 2.5% increment in Trial I; 1.25–7.50% BW day^?1 with a 1.25% increment in Trial II; and 2.0–7.0% BW day^?1 with a 1.0% increment in Trials III and IV. Four replicates with 50 fry per tank in Trials I‐III and 30 fry per tank in Trial IV were assigned randomly to each feeding rates. The final body weight, specific growth rate, feed efficiency, protein retention, and whole‐body moisture, lipid, and energy contents were significantly (P < 0.05) affected by the feeding rates. The optimum feeding rates determined by the broken‐line model were 7.1, 5.7 and 5.3% BW day^?1 for Trials I, II and IV, when the fry were 5, 6 and 8 weeks after their initiation of exogenous feeding, respectively.     

Effect of different biofloc system on water quality,biofloc composition and growth performance in Litopenaeus vannamei (Boone, 1931)

The experiment was conducted with three biofloc treatments and one control in triplicate in 500 L capacity indoor tanks. Biofloc tanks, filled with 350 L of water, were fed with sugarcane molasses (BFT_S), tapioca flour (BFT_T), wheat flour (BFT_W) and clean water as control without biofloc and allowed to stand for 30 days. The postlarvae of Litopenaeus vannamei (Boone, 1931) with an Average body weight of 0.15 ± 0.02 g were stocked at the rate of 130 PL m^?2 and cultured for a period of 60 days fed with pelleted feed at the rate of 1.5% of biomass. The total suspended solids (TSS) level was maintained at around 500 mg L^?1 in BFT tanks. The addition of carbohydrate significantly reduced the total ammonia‐N (TAN), nitrite‐N and nitrate‐N in water and it significantly increased the total heterotrophic bacteria (THB) population in the biofloc treatments. There was a significant difference in the final average body weight (8.49 ± 0.09 g) in the wheat flour treatment (BFT_W) than those treatment and control group of the shrimp. Survival of the shrimps was not affected by the treatments and ranged between 82.02% and 90.3%. The proximate and chemical composition of biofloc and proximate composition of the shrimp was significantly different between the biofloc treatments and control. Tintinids, ciliates, copepods, cyanobacteria and nematodes were identified in all the biofloc treatments, nematodes being the most dominant group of organisms in the biofloc. It could be concluded that the use of wheat flour (BFT_W) effectively enhanced the biofloc production and contributed towards better water quality which resulted in higher production of shrimp.     

Mortality and pathology of hybrid catfish,Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), associated with Edwardsiella ictaluri infection in southern Thailand

Enteric septicaemia of catfish (ESC) caused by Edwardsiella ictaluri is becoming an increasing problem in aquaculture and has been reported worldwide in a variety of fish species. This study reports ESC in hybrid catfish, Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), cultured in southern Thailand. The bacteria were identified as E. ictaluri by conventional and rapid identification systems, as well as by genetic and phylogenetic characterization. Analysis of 16S rRNA indicated 100% homology to the 16S rRNA sequence of several E. ictaluri strains in GenBank. Plasmid profiles demonstrated 4.0‐ and 5.6‐kb plasmids, compared with the 4.8‐ and 5.6‐kb plasmids in the US isolates, and representative genes of three of the four known pathogenicity islands of US isolates were present. Serologically, lipopolysaccharide (LPS) purified from the Thai isolates was not recognized by a monoclonal antibody against the LPS of US isolates. Fish experimentally infected with E. ictaluri showed 23–100% mortality within 14 days with a 168‐h LD₅₀ of 6.92 × 10⁷ CFU mL^?1 by immersion and a 96‐h LD₅₀ of 1.58 × 10⁶ CFU fish^?1 by intraperitoneal injection. Examination of tissue sections obtained from both naturally and experimentally infected fish indicated that infection of hybrid catfish with E. ictaluri produced lesions in several organs including liver, kidney, spleen, heart and brain. Histopathology findings included cellular necrosis, focal haemorrhage, infiltration of lymphocytes and multifocal granulomatous inflammation in the infected organs.     

In vivo evaluation of the effects of commercial Bacillus probiotics on survival and development of Litopenaeus vannamei larvae during the early hatchery period

Three in vivo experiments were conducted to measure the effectiveness of commercial Bacillus probiotics on survival and development of Litopenaeus vannamei larvae from nauplii 3–4 (N_3–4) to zoea 3 (Z₃) stages. Experiment I: Nine commercial Bacillus probiotics were individually added to larvae twice at N₆ and Z₁ at levels of 2, 20, and 100 mg L^?1. Only six of nine products at 20 mg L^?1 exhibited higher or significantly higher larval survival (P < 0.05) than the control. Experiment II: Two superior products from the first experiment were administered to larvae with six dose frequencies at 20 mg L^?1. For both products, three doses, once at each of N_3–4, N₆ and Z₁ stages, yielded the best larval survival and development rates (P < 0.05), and these were confirmed by enhanced activities of tryptase and amylase. Experiment III: The isolates of these two products, identified as Bacillus subtilis and Bacillus licheniformis, were delivered to larvae singly at concentrations of 1.0 × 10⁸ and 1.0 × 10⁹ CFU L^?1, or at the same concentrations by mixing the two equally. At 1.0 × 10⁹ CFU L^?1, B. subtilis exerted more beneficial effects on larvae than B. licheniformis or the mixture. Therefore, the optimal dosage and dose frequency of commercial Bacillus products should be evaluated prior to large‐scale application in shrimp hatcheries to avoid futility or even adverse effects, as spore counts and non‐bacterial ingredients are key factors influencing the efficacy of Bacillus probiotics.     

Studies on bacterial pathogens isolated from diseased torafugu (Takifugu rubripes) cultured in marine industrial recirculation aquaculture system in Shandong Province,China

In spring of 2011, an epidemic outbreak of torafugu with high mortality occurred in an aquafarm with marine industrial recirculation aquaculture system (MIRAS) in Yantai, Shandong Province, China. The diseased fish showed anorexia, haemorrhaging and festering fin and skin and swelling internal organs. Forty‐five dominant bacterial isolates were obtained from the diseased fish, and were found to belong to 12 species according to 16S rRNA gene sequences. One strain from each species was selected to test the pathogenicity, and five strains were showed to be virulent to zebrafish. Whereas Enterovibrio nigricans Fr42 was highly virulent with the LD₅₀ of 7.8 × 10⁴ CFU g^?1, Photobacterium swingsii Fr23, Vibrio owensii Fr40, V. harveyi Fr51 and V. rotiferianus Fr71 were moderately virulent with the LD₅₀ of 1.7 × 10⁶ to 8.4 × 10⁶ CFU g^?1. Both the bacteria and their extracellular products of the five strains were found to show phospholipase, caseinase, gelatinase, amylase and/or lipase activities. The production of N‐acyl homoserine lactones (AHLs) of the five strains was detected by three different AHLs biosensors, and three of them were found to produce AHLs by at least one kind of biosensor. This is the first study describing various opportunistic bacterial pathogens of fish cultured in MIRAS in China.     

Evaluation of live duckweed,Wolffia globosa as an allochthonous feed for Labeo rohita fry during nursery rearing

A 40 days feeding trial with Labeo rohita (rohu) fry was conducted in six outdoor cemented tanks (5m × 4m × 1 m; 6–8 cm soil base) to establish the relative impact of on‐farm grown live Wolffia globosa (a duckweed with crude protein of 28.57%) and a formulated iso‐nitrogenous feed (crude protein 28.37%) on growth performance, survival, health and overall quality of fingerlings. The experimental tanks were dried, limed at 250 kg/ha, filled with ground water and subsequently applied with cow dung (soaked overnight) and mustard oil cake at 1,000 kg/ha and 50 kg/ha respectively. The fry were stocked at 0.3 millions/ha in six prepared tanks, and fed live Wolffia (T1) and formulated feed (T2). Results of the study indicate better performance (p < 0.05) of fry in terms of final mean weight (7.83 g), length (8.82 cm), daily growth index (6.51 g %/day) and survival (89.5%) than those fed on formulated feed. Furthermore, significantly improved (p < 0.05) feed utilization parameters viz. apparent feed conversion ratio (AFCR), apparent protein conversion efficiency (APCE), apparent protein efficiency ratio (APER); haematological parameters viz. total erythrocyte count (TEC), total leucocyte count (TLC), packed cell volume (PCV), plasma glucose and protein of fry were also observed in T1 (live Wolffia). Overall findings from this study hints towards the possible utilization of on‐farm grown live Wolffia as a suitable nutrient source for fry rearing of L. rohita.