The effects of a prolonged photoperiod and light source on growth,sexual maturation,fin condition,and vulnerability to fungal disease in brook trout Salvelinus fontinalis

The effects of an artificially prolonged photoperiod on growth, survival rate, colouration, and sexual maturation in brook trout Salvelinus fontinalis during pre‐ and post‐spawning periods from 21 June to 06 November were investigated. Fish of mean initial weight ?150 g were reared at ambient photoperiod as well as with an artificially prolonged photoperiod produced by either a light‐emitting diode or a metal‐halide light. The fish groups subjected to a prolonged period of artificial light grew significantly larger and had a higher survival rate (p < 0.05), regardless of sex, and showed lower occurrence of fungal disease compared to controls reared in the natural photoperiod. We found a significantly higher number of sexually mature fish in the control groups compared with experimental groups. The increased photoperiod effectively delayed gonad development and increased somatic growth in both male and female brook trout, and also increased resistance to fungal disease. The increased photoperiod produced an observable difference in fish colouration, with control groups exhibiting more intensive spawning colouration.     

The effect of timing of extended photoperiod on growth and maturity of brook trout (Salvelinus fontinalis)

The influence of timing of extended photoperiods on growth and maturity of brook trout was investigated in a 112‐day experiment. The fish with mean initial weight of ~192 g were reared under four light regimes: one control group with natural ambient photoperiod and three groups exposed to an 18L:6D regime initiated at days 1, 23 or 46 of the growth trial. Light‐emitting diodes, with intensity of 250–1000 lux, depending on the distance from the light source, were used for extending light periods. There was a positive effect of prolonged day length on fish growth (p < 0.05), and a delay in gonad development and sexual maturity. Significantly higher numbers of sexually mature fish were found among controls groups, regardless of sex. Survival rate was not affected by light regime. This study demonstrated that the short‐term expansion of the photo period delayed maturation and increased the growth rate of brook trout.     

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

Erythromycin and florfenicol treatment of rainbow trout Oncorhynchus mykiss (Walbaum) experimentally infected with Flavobacterium psychrophilum

Flavobacterium psychrophilum is responsible for significant economic losses in rainbow trout aquaculture. Antimicrobial treatment remains the primary means of control; however, there are limited choices available for use. The objectives of the study were therefore to determine the minimum inhibitory concentrations for erythromycin and florfenicol in selected F. psychrophilum isolates and to evaluate their clinical treatment efficacy in experimentally infected rainbow trout. All isolates tested had moderate susceptibility to florfenicol and erythromycin except one isolate, which had low susceptibility to erythromycin. Two isolates (one with moderate and one with low susceptibility to erythromycin) were used in an experimental infection trial. Rainbow trout juveniles were injected intraperitoneally with 10⁸ cfu/fish and after mortality had begun, fish were given erythromycin‐ and florfenicol‐medicated feed at a rate of 75 mg kg^?1 day^?1 and 10 mg kg^?1 day^?1 fish body weight, respectively, for 10 consecutive days. The splenic F. psychrophilum load was determined using an rpoC quantitative PCR throughout the 30‐day trial. Relative to antibiotic‐free controls, erythromycin treatment significantly (p < 0.05) reduced mortality of rainbow trout juveniles infected with FPG101, even when treatment was initiated after clinical signs developed.     

Physiological differences between gametophytes and sporophytes of five cultivar strains of Saccharina japonica

The demand for high‐yield cultivars of Saccharina japonica in farming areas calls for a practical and simple method to select desired parental plants for seed breeding. In this study, we evaluated PSII photosynthetic performance by measuring chlorophyll fluorescence and the nutrient uptake rate in both the micro‐gametophyte and macro‐young‐sporophyte in five native strains (S1, S2, S3, S4 and S5) of S. japonica with different morphological characteristics. We found significant differences in PSII photosynthetic parameters and nitrate uptake rates between the strains studied in both gametophytes and young sporophytes. Female gametophytes of all strains presented larger cell sizes than males (P < 0.05). Among all tested strains, S2 presented the largest cell sizes and highest levels of PSII photosynthetic parameters and nutrient uptake rates in female gametophytes and young sporophytes. In open sea cultivation, S2 presented the greatest width and largest fresh weight values but the lowest thickness among all strains studied. Correlation analysis further indicated that the algal fresh weight showed strong positive relationships with cell size (R² > 0.747), the PSII photosynthetic parameters of female gametophytes (R² > 0.689) and the nitrate uptake rate of young sporophytes (R² = 0.640). These findings are considered promising for the future seed breeding of renewable, high‐yield strains.     

Effects of rearing density on growth,fatty acid profile and bioremediation ability of polychaete Nereis diversicolor in an integrated aquaculture system with rainbow trout (Oncorhynchus mykiss)

The objective of this study was to assess the effects of rearing density on the growth and fatty acid profile of Nereis diversicolor and on its capability to bioremediate wastewater in an integrated culture system with rainbow trout (Oncorhynchus mykiss). To this end, a batch of juvenile N. diversicolor (0.03 ± 0.01 mg) was assigned into four different densities (T₁ = 250, T₂ = 500, T₃ = 1,000, T₄ = 2,000, besides T₀ = with no worm) in three replicates. The worm groups were fed with solid waste that was supplied from tanks containing rainbow trout (107.17 ± 13.5 g; 1.39 ± 0.18 kg/m³). During the experiment (60 days), the water temperature was 17.71 ± 0.6°C. The results revealed that SR%, SGR% and WG% of N. diversicolor in T₁ were significantly higher than those of T₂, T₃ and T₄. Both FER rate and biomass gain in T₄ were significantly higher than those in the other groups. The highest removal rate of NO₂‐N (73.72%), NH₃‐N (65.70%), PO₄‐P (59.32%), BOD₅ (69.60%) and TSS (82.33%) were observed in T₄. The major fatty acids presents were palmitic acid, oleic acid, linoleic acid, stearic acid and alpha‐linolenic acid in all worm‐treated groups, with no difference observed in the concentration of these fatty acids among them. Taken together, these findings suggest that organic wastes from rainbow trout farms could be recycled to achieve a sustainable aquaculture goal, and demonstrate that a high percentage of fatty acids in fish feed is not absorbed by rainbow trout but is done by N. diversicolor.     

Virulence of Flavobacterium psychrophilum isolates in rainbow trout Oncorhynchus mykiss (Walbaum)

Flavobacterium psychrophilum, the causative agent of bacterial cold‐water disease (BCWD) in freshwater‐reared salmonids, is also a common commensal organism of healthy fish. The virulence potential of F. psychrophilum isolates obtained from BCWD cases in Ontario between 1994 and 2009 was evaluated. In preliminary infection trials of rainbow trout juveniles, significant differences (0% to 63% mortality) in the virulence of the 22 isolates tested were noted following intraperitoneal injection with 10⁸ cfu/fish. A highly virulent strain, FPG 101, was selected for further study. When fish were injected intraperitoneally with a 10⁶, 10⁷ or 10⁸ cfu/fish of F. psychrophilum FPG 101, the 10⁸ cfu/fish dose produced significantly greater mortality (p < 0.05). The bacterial load in spleen samples collected from fish every 3 days after infection was determined using rpoC quantitative polymerase chain reaction amplification and by plate counting. Bacterial culture and rpoC qPCR were highly correlated (R² = 0.92); however, culture was more sensitive than the qPCR assay for the detection of F. psychrophilum in spleen tissue. Ninety‐seven per cent of the asymptomatic and the morbid fish had splenic bacterial loads of <2.8 log₁₀ gene/copies and >3.0 log₁₀ gene copies/reaction, respectively, following infection with 10⁸ cfu/fish.     

Review of molluscan bivalve condition index calculations and application in Northern Quahogs Mercenaria mercenaria

Condition index (CI) is a widely used parameter in ecological and physiological studies. For molluscan bivalves, no universal consensus exists for CI calculation. A literature review revealed that a total of 19 methods have been used for CI calculation in molluscan bivalves. To evaluate the CI calculation in northern quahogs Mercenaria mercenaria (also called hard clams), market‐sized clams at 12–14 months old (n = 300 from three populations) were collected for gravimetrical and volumetric measurements. Significant correlations were identified among body sizes (body length, height and width), body weights (total weight, wet meat weight, dry meat weight, wet shell weight, dry shell weight) and body volume (p < .0001, R values ≥ .800). Referencing previously used CI equations for molluscan bivalves, a total of 18 equations were evaluated the suitability, considering non‐lethal measurements, for northern quahogs through correlation analysis of the CIs from different equations. With equations constructed as the percentages of dry or wet meat weight out of total weight, dry shell weight, dry shell plus meat weight, body volume, body length*height*width, shell length or cubic of shell length, CIs were significantly correlated, meaning that these equations were exchangeable. Specifically, three equations with non‐lethal measurements (percentages of total body weight out of total body volume, body length*height*width or cubic of body length) were evaluated effective. Overall, this publication provided a review for CI calculations in molluscan bivalves and evaluated the CI equations for northern quahogs.     

Fecundity and egg dimension of freshwater prawn Macrobrachium assamense peninsulare (Tiwari, 1958) from Rawasan stream of Garhwal,Central Himalaya,India

The population of the freshwater animal was declined day by day due to the degradation of habitat and use of illegal fishing methods, which cause a great threat to these aquatic animals. The main purpose of this study was to scrutinize the wild population and fecundity of freshwater prawn Macrobrachium assamense peninsulare in Rawasan stream to assess the health status of prawn. Fecundity was estimated by a random sampling of prawn from five selected sites of Rawasan stream. A wild collection of a total of 35 ovigerous females was collected for an interval of 2 years (August 2013–July 2015) from May to September month, that is breeding session. The maximum number of eggs was 102 eggs in an individual having 52 mm total length and 2.66 g body weight. It was concluded that fecundity of Macrobrachium assamense peninsulare was low, and egg dimension was larger than other closely related species of freshwater prawn. Fecundity of Macrobrachium assamense peninsulare showed a positive relationship with a total weight (R² = .71) and moderate with the total length (R² = .38). A strong correlation exists between egg mass and total weight, and between egg mass and total length of female (R² = .849 and R² = .77 at p < .05) respectively. The numbers of eggs, that is fecundity, in a female were increased with the body growth of an individual, which results in a linear graph.     

First identification and characterization of Streptococcus iniae obtained from tilapia (Oreochromis aureus) farmed in Mexico

This is the first study to isolate, identify and characterize Streptococcus iniae as the causative disease agent in two tilapia (Oreochromis aureus) populations. The populations were geographically isolated, of distinct origins, and did not share water sources. Affected fish showed various external (e.g., exophthalmia and cachexia, among others) and internal (e.g., granulomatous septicaemia and interstitial nephritis, among others) signs. All internal organ samples produced pure cultures, two of which (one from each farm, termed S‐1 and S‐2) were subjected to biochemical, PCR and 16S rRNA sequencing (99.5% similarity) analyses, confirming S. iniae identification. The two isolates presented genetic homogeneity regardless of technique (i.e., RAPD, REP‐PCR and ERIC‐PCR analyses). Pathogenic potentials were assessed through intraperitoneal injection challenges in rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). Rainbow trout mortalities were respectively 40% and 70% at 10⁴ and 10⁶ CFU per fish with the S‐1 isolate, while 100% mortality rates were recorded in zebrafish at 10² and 10⁴ CFU per fish with the S‐2 isolate. The obtained data clearly indicate a relationship between intensified aquaculture activities in Mexico and new disease appearances. Future studies should establish clinical significances for the tilapia industry.     

Efficacy of a polyvalent injectable vaccine against Flavobacterium psychrophilum administered to rainbow trout (Oncorhynchus mykiss L.)

Flavobacterium psychrophilum is one of the most important pathogens affecting cultured rainbow trout (Oncorhynchus mykiss). Recent information from UK salmonid farms showed country‐wide distribution of genetically and serologically divergent clones, which has hampered the development of a vaccine for rainbow trout fry syndrome. The current study assessed the efficacy of an injectable polyvalent vaccine containing formalin‐inactivated F. psychrophilum in rainbow trout. The vaccine was formulated with an oil adjuvant (Montanide ISA 760VG) or formalin‐killed cells alone. Duplicate groups of trout (60 ± 13 g) were given phosphate‐buffered saline or vaccine formulated with Montanide by intra‐peritoneal (i.p.) injection and challenged by intra‐muscular (i.m.) injection with a homologous and a heterologous isolate of F. psychrophilum at 525 degree days post‐vaccination (dd pv). Significant protection was achieved in vaccinated fish (p = 0.0001, RPS 76% homologous, 88% heterologous). Efficacy of the adjuvanted vaccine was also demonstrated by heterologous challenge at 1155 dd pv resulting in 100% protection, whereas survival in the un‐adjuvanted group was not significantly different from control fish. Levels of specific antibody at 1155 dd pv, as measured by ELISA, were significantly higher in the fish vaccinated with adjuvant when compared with unvaccinated fish.     

Suppression of invasive lake trout in an isolated backcountry lake in Glacier National Park

Fisheries managers have implemented suppression programmes to control non‐native lake trout, Salvelinus namaycush (Walbaum), in several lakes throughout the western United States. This study determined the feasibility of experimentally suppressing lake trout using gillnets in an isolated backcountry lake in Glacier National Park, Montana, USA, for the conservation of threatened bull trout, Salvelinus confluentus (Suckley). The demographics of the lake trout population during suppression (2009–2013) were described, and those data were used to assess the effects of suppression scenarios on population growth rate (λ) using an age‐structured population model. Model simulations indicated that the population was growing exponentially (λ = 1.23, 95% CI: 1.16–1.28) prior to suppression. However, suppression resulted in declining λ (0.61–0.79) for lake trout, which was concomitant with stable bull trout adult abundances. Continued suppression at or above observed exploitation levels is needed to ensure continued population declines.     

A new potential therapeutic remedy against Aeromonas hydrophila infection in rainbow trout (Oncorhynchus mykiss) using tetra,Cotinus coggygria

Different antibiotic‐based drugs are being used for the treatment of Aeromonas hydrophila infection in rainbow trout, and several studies emphasize the use of medicinal plants as immunostimulants for prophylactic measure against Aeromoniasis disease. However, therapeutic effects of aqueous methanolic extracts of tetra (Cotinus coggygria) against A. hydrophila in rainbow trout were not investigated. Four different concentrations of tetra extract (0 [control], 4, 8 and 12 mg/100 µl) and also two different positive control groups (florfenicol and doxycycline antibiotics) were administered orally using feeding needles to individual rainbow trout, Oncorhynchus mykiss of all experimental groups twice a day after intramuscular inoculation of A. hydrophila. The study period was for 10 days. On 0th, 3rd, 7th and 10th day, blood and tissues were collected from the fish and changes in humoral immune responses, haematology and immune‐related gene expressions were determined. In the study, superoxide radical production was decreased generally in all experimental groups except in 12 mg tetra and florfenicol treatments compared to control (p < .05). Lysozyme activity was generally decreased (p < .05), or no differences were observed in all experimental groups compared to the control. Myeloperoxidase activity was significantly increased in florfenicol‐treated fish group on 7th day (p < .05). Generally, myeloperoxidase activity showed an increase in almost all tetra‐treated groups. Haematological parameters increased but were not significantly high enough in treatments. Almost all immune‐related gene expressions were significantly enhanced on 3rd and 10th day of the study. Survival rate of 53.33% was found in control group. There were no significant differences in survival between control and 4 mg tetra‐treated group (p > .05). All the other groups' survival rate was significantly increased compared to control. The highest survival rate was found in florfenicol group (80%). In 12 mg tetra‐, doxycycline‐ and 8 mg tetra‐treated groups, survival rate was recorded as 74.44%, 70% and 70%, respectively. Our results suggest that tetra methanolic extract is an effective therapeutic remedy against A. hydrophila infection in rainbow trout at the dose of 24 mg/32.34 g body weight/day.     

Effects of dietary starch levels on the growth,plasma metabolites and expression of genes involved in glucose transport and metabolism in Pelodiscus sinensis juveniles

A 10‐week feeding trial was conducted to evaluate the growth performance, glucose transport and metabolism of Chinese soft‐shelled turtles (Pelodiscus sinensis) exposure to graded levels of dietary starch (0.52%, 7.43%, 14.74%, 22.99% and 31.38%). The 360 turtles (initial body weight, 12.94 ± 0.50 g) with 12 replicates were randomly assigned to five experimental diets. The highest weight gain and specific growth rate (SGR) were observed in 7.43% group and the lowest in 31.38% group. The protein efficiency ratio, whole‐body lipid contents, hepatic glycogen contents and the 4‐hr postprandial plasma glucose levels were significantly increased with the increment of starch levels (p < .05). In contrast, the daily feed intake and feed conversion ration were significantly declined (p < .05). The mRNA levels of glucose transporter 2, glucokinase, pyruvate kinase, malic enzyme and acetyl‐CoA carboxylase alpha genes in the liver significantly increased as the increase in starch levels at 4‐hr and 24‐hr post feeding (p < .05). No significant differences were observed in the expression of gluconeogenesis genes at each time point (p > .05). These results suggested that dietary addition of starch up‐regulated hepatic glycolysis, glycogenesis and lipogenesis genes expression, but the deficient response of gluconeogenesis to dietary starch might be part of the causes limited the starch utilization. Based on the secondary polynomial regression of SGR, y = ?0.0011x² + 0.028x + 1.63 (R² = 0.9292), the 12.73% inclusion level of dietary starch was recommended in juvenile turtles.     

Evaluation of brook trout production in a coldwater recycle aquaculture system

Brook trout (Salvelinus fontinalis) are a commercially important coldwater species reared in Wisconsin and the Midwestern United States. Brook trout are raised by private, tribal, state, and federal fish hatchery facilities in Wisconsin. Approximately 10% of private coldwater aquaculture operations are presently raising brook trout of various strains for stocking uses and a limited amount for food markets. Growing brook trout to a larger size, if they can be reared in a shorter time span, may present a potential new sector for the aquaculture market in the Midwestern US. The present study reports hatchery production attributes, i.e., growth, survival, fin condition, feed efficiency, water chemistry requirements and general husbandry of Lake Superior strain (Nipigon) brook trout reared in a recirculating aquaculture system (RAS), operated at an average temperature of 13 °C. The recycle system at NADF reared 1379 kg of brook trout over a 10-month period from fingerling (9 g) to market size (340–454 g). The trout grew faster (0.84 g/day and 0.64 mm/day) in the RAS than fish cultured in traditional flow-through tank culture utilizing ground water at 7.6 °C (0.14 g/day and 0.35 mm/day). Final average weight of RAS fish was 260 g, while the flow-through fish averaged 65 g. Final tank densities for the RAS averaged 40.4 kg/m³ while flow-through tanks averaged 31.2 kg/m³. Throughout the project, feed conversions in the RAS ranged from 0.9 to 1.3. Water quality variables such as TAN, nitrite, DO, temperature, TSS, CO₂, ph, etc. were within safe limits for brook trout and will be discussed. It does appear from this initial research project that market size brook trout can be raised successfully in a recycle system within a similar time frame as a rainbow trout produced in a Wisconsin typical flow-through facility.     

Dietary protein requirement of juvenile obscure puffer,Takifugu obscurus

A 10‐week feeding experiment was conducted to determine the optimum dietary protein requirement of juvenile obscure puffer (Takifugu obscurus). Six isoenergetic (20 MJ kg^?1 gross energy) diets were formulated to contain graded levels of 34%, 38%, 42%, 46%, 50% or 54% crude protein (as dry matter basis). The results showed final body weight, weight gain and specific growth rate (SGR) increased significantly with increasing protein levels up to 42% and then decreased thereafer. Second‐order polynomial regression analysis (y = ?0.0024x² + 0.1788x ? 1.3196, R² = 0.9032) indicated a maximum SGR at protein level of 37%. Feed conversion ratio (FCR) decreased with increasing levels of dietary protein up to 42% and increased thereafter. Second‐order polynomial regression analysis (y = 0.0054x² ? 0.4351x + 10.391, R² = 0.753) indicated a minimum FCR at protein level of 40%. Protein efficiency ratio (PER) of fish fed the 34%, 38% and 42% diets was significantly higher than that of fish fed the 46%, 50% and 54% diets, and broken‐line analysis indicated PER tended to decrease when dietary protein level was higher than 40%. Generally, whole body lipid content, total cholesterol, low‐density lipoprotein cholesterol and triacylglycerol decreased with increasing levels of dietary protein. Fish fed the 42% protein diet showed the highest essential amino acids (histidine, isoleucine, leucine, lysine and threonine) and non‐essential amino acids (aspartic acid and glutamic acid) in muscle. Based on the second‐degree polynomial regression analysis of SGR and FCR and broken‐line analysis of PER, the optimal dietary protein level of obscure puffer is estimated to be between 37% and 40% (% as dry matter basis).     

Detection and quantification of Aeromonas schubertii in Channa maculata by TaqMan MGB probe fluorescence real‐time quantitative PCR

Aeromonas schubertii is a major epidemiological agent that threatens cultured snakeheads (Channidae) and has caused great economic losses in fish‐farming industries in China in recent years. In present study, a specific TaqMan minor groove binder (MGB) probe fluorescence real‐time quantitative PCR (qPCR) assay was developed to rapidly detect and quantify A. schubertii. A pair of qPCR primers and a TaqMan MGB probe were selected from the rpoD gene, which were shown to be specific for A. schubertii. A high correlation coefficient (R² = 0.9998) in a standard curve with a 103% efficiency was obtained. Moreover, the qPCR method's detection limit was as low as 18 copies/μl, which was 100 times more sensitive than that of conventional PCR. The detection results for the A. schubertii in pond water and fish tissue were consistent with those of the viable counts. Bacterial load changes detected by qPCR in different tissues of snakeheads infected with A. schubertii showed that the gills and intestines may be the entry for A. schubertii, and the spleen and kidney are major sites for A. schubertii replication. The established method in present study should be a useful tool for the early surveillance and quantitation of A. schubertii.     

Assessment of cross‐protection to heterologous strains of Flavobacterium psychrophilum following vaccination with a live‐attenuated coldwater disease immersion vaccine

Bacterial coldwater disease, caused by Flavobacterium psychrophilum, remains one of the most significant bacterial diseases of salmonids worldwide. A previously developed and reported live‐attenuated immersion vaccine (F. psychrophilum; B.17‐ILM) has been shown to confer significant protection to salmonids. To further characterize this vaccine, a series of experiments were carried out to determine the cross‐protective efficacy of this B.17‐ILM vaccine against 9 F. psychrophilum isolates (representing seven sequence types/three clonal complexes as determined by multilocus sequence typing) in comparison with a wild‐type virulent strain, CSF‐259‐93. To assess protection, 28‐day experimental challenges of rainbow trout (Oncorhynchus mykiss) fry were conducted following immersion vaccinations with the B.17‐ILM vaccine. F. psychrophilum strains used in challenge trials were isolated from several fish species across the globe; however, all were found to be virulent in rainbow trout. The B.17‐ILM vaccine provided significant protection against all strains, with relative percent survival values ranging from 51% to 72%. All vaccinated fish developed an adaptive immune response (as measured by F. psychrophilum‐specific antibodies) that increased out to the time of challenge (8 weeks postimmunization). Previous studies have confirmed that antibody plays an important role in protection against F. psychrophilum challenge; therefore, specific antibodies to the B.17‐ILM vaccine strain appear to contribute to the cross‐protection observed to heterologous strain. The ability of such antibodies to bind to similar antigenic regions for all strains was confirmed by western blot analyses. Results presented here support the practical application of this live‐attenuated vaccine, and suggest that it will be efficacious even in aquaculture operations affected by diverse strains of F. psychrophilum.     

Optimization of de‐oiled rice bran inclusion level in the diet of Labeo rohita: Effect on growth performance,nutrient utilization,insulin‐like growth factors I and II gene expression

A 60‐day feeding trial was conducted to optimize the inclusion level of de‐oiled rice bran (DORB) in the diet of Labeo rohita fingerlings. Six iso‐nitrogenous (260g/kg), iso‐lipidic (70g/kg) and iso‐energetic (15 MJ/Kg) diets were prepared with different inclusion level of DORB viz., T33, T38_, T43, T48, T53 and T58. One hundred and eighty fingerlings (9.8 ± 0.5 g) were randomly distributed in seven experimental groups in triplicates following a completely randomized design. The weight gain %, specific growth rate, protein efficiency ratio and feed efficiency ratio were found similar in 33%, 38% and 43% DORB fed groups. The expression of both insulin‐like growth factors I and II (IGF‐I and II) was significantly (p < 0.05) higher in T33, T38 and T43 groups. Based on second order polynomial regression analysis (y = ?0.0059x² + 0.4573x ? 4.8404, R² = 0.95), the expression of IGF‐I was found to be maximum at 39.5% inclusion level of DORB. Significantly (p < 0.05) lower muscle RNA content, RNA‐DNA ratio and higher feed conversion ratio were found in T58 group. Hence, from these results, it can be concluded that the optimum inclusion of DORB in the diet (260g/kg CP and 70g/kg lipid) of L. rohita should be 39.5% to support maximum nutrient utilization and growth performance.     

Effect of combination of dietary Bacillus subtilis and trans‐cinnamic acid on innate immune responses and resistance of rainbow trout,Oncorhynchus mykiss to Yersinia ruckeri

The present study investigated the effects of combination of dietary Bacillus subtilis and trans‐cinnamic acid on serum biochemical parameters, innate immune responses and resistance of rainbow trout, Oncorhynchus mykiss to Yersinia ruckeri. Six experimental groups of fish with mean weights of 20.58 ± 0.35 g were used in the study. Five experimental groups of fish were fed diets containing Bacillus subtilis (10⁷ per gram) or a mix of the Bacillus subtilis (BS) and trans‐cinnamic acid (25 mg/kg‐25trcBS, 50 mg/kg‐50trcBS, 75 mg/kg‐75 trcBS, 150 mg/kg‐150 trcBS), whereas an additive‐free basal diet served as the control (Cont). In this study, an increase was observed in granulocyte percentage, respiratory burst activity, phagocytic activity, phagocytic index, myeloperoxidase activity and total antiprotease activity especially in fish fed with mix of the BS and trans‐cinnamic acid‐supplemented diets (p < .05). Moreover, at the end of the 20‐day challenge period the survival rates and antibody titre (p < .05), and relative per cent survival were higher in the BS group and all trcBS groups compared with control group. As a conclusion, the results in the present study show that feeding rainbow trout with diets containing a mix of B. subtilis and trans‐cinnamic acid over a 60‐day period might be sufficient for improving fish immune responses and disease resistance against Y. ruckeri.