Dietary supplementation of methionine, lysine, and tryptophan as possible modulators of growth, immune response, and disease resistance in striped catfish (Pangasius hypophthalmus).

The present study investigated the potential role of different essential amino acids (AA) in striped catfish (Pangasius hypophthalmus). Fish (initial weight = 17.91±0.27 g, n = 260) were fed with eight isonitrogenous (30%), and isolipidic diets (6%) formulated to include different combinations of tryptophan (Trp), methionine (Met), and lysine (Lys) (T0: Zero AA, T1: Trp, T2: Lys, T3: Met, T4: Trp+Met, T5: Lys+Trp, T6: Met+Lys, T7: Lys+Trp+Met) for eight weeks. The dose of amino acid supplementation, whether individually or in combination, was 5g of each amino acid per kg of diet. The trial comprised eight treatments, with each treatment consisted of three replicates (n = 10/replicate). At the end of the growth experiment, the highest total body weight, crude protein, digestive enzymatic activity, immune response, and amino acids level were observed in treatments supplemented with amino acids compared to T0. After the growth experiment, fish in all treatments were exposed to Staphylococcus aureus (5×105 CFU/ml). For bacterial challenge trial, the T0 treatment was designated as positive (+ve T0) and negative control (-ve T0). Following the S. aureus challenge, fish fed with amino acids showed a better response to reactive oxygen species and lipid peroxidation, as indicated by the increased levels of catalase and superoxide dismutase. Conversely, the concentration of malondialdehyde gradually decreased in all treatments compared to the +ve T0 treatment. It is concluded that supplementation of amino acids improved the growth, protein content, and immunocompetency against S. aureus in striped catfish. The most favorable outcomes in striped catfish were shown by fish supplemented with T7 diet. These essential amino acids hold potential as efficient supplements for use in the intensive aquaculture for striped catfish.

Effects of Bacillus subtilis as a single strain probiotic on growth, disease resistance and immune response of striped catfish (Pangasius hypophthalmus).

The present study investigated the potential role of Bacillus subtilis as probiotic in striped catfish (Pangasius hypophthalmus). Fish (initial weight = 150.00±2.63g n = 180) were stocked in circular tanks. Four isonitrogenous (30%) and isolipidic (3.29%) diets were formulated having supplementation of B. subtilis at four different levels (P0; 0, P1: 1×106, P2: 1×108 and P3: 1×1010 CFU/g). Each treatment had three replicates, while each replicate had fifteen fish. The trial started on second week of July and continued for eight weeks. Growth, feed conversion ratio, crude protein content, the concentration of amylase and protease, the profile of both dispensable and non-dispensable amino acids in all four dietary groups increased with a gradual increase of B. subtilis in the diet. At the end of growth experiment, fish in all four groups were exposed to Staphylococcus aureus (5×105 CFU/ml). After S. aureus challenge, fish fed with B. subtilis responded better to damage caused by reactive oxygen species and lipid peroxidation and better survival rate. The catalase and superoxide dismutase level also increased in response to bacterial challenge in B. subtilis fed groups. On the other hand, the concentration of malondialdehyde gradually decreased in these groups (+ve P0 >P1>P2>P3). It is concluded that supplementation of B. subtilis as a probiotic improved the growth, protein content, antioxidant response and immunocompetency against S. aureus in striped catfish. The optimum dosage of B. subtilis, at a concentration of 1×1010 CFU/g, resulted in the most favorable outcomes in striped catfish. This single bacterial strain can be used as an effective probiotic in large scale production of aquafeed for striped catfish. Future studies can investigate this probiotic's impact in the intensive culture of the same species.

Dietary replacement of soybean meal with black soldier fly larvae meal in juvenile Labeo rohita and Catla catla: Effects on growth, nutritional quality, oxidative stress biomarkers and disease resistance.

This experiment aimed to investigate the effects of partial substitution of crude protein from soybean meal (SBM) with black soldier fly (Hermetia illucens) larvae meal (BSFLM) in juvenile rohu (Labeo rohita) and catla (Catla catla). Four isonitrogenous diets (23% crude protein) were formulated to replace 0% (T0), 40% (T40), 80% (T80) and 100% (T100) crude protein from SBM with BSFLM. Triplicate groups of each species (10 fish per replicate) were fed in an eight week growth experiment. After final sampling (n = 20 fish per dietary group), the remaining fish were exposed to bacterial (Staphylococcus aureus) challenge (0.80 CFU/ml) for 15 days. Rohu fed with BSFLM substituted diets showed significantly higher growth and feed conversion ratio as compared to those in T0. Catla fed with BSFLM substituted diets showed slightly higher growth indices. The growth response of rohu to BSFLM substitution was better than that noted in catla in all groups. The chemical composition, amino acids and fatty acids profile, haematological and biochemical parameters, levels of liver function enzymes measured in T0, T40, T80 and T100 were similar between four dietary groups in both species. However, the maximum value of cholesterol and triglycerides were noted in T100 both in catla and rohu. The values of lauric acid, α-linolenic acid, decosahexanoic acid, n3:n6 fatty acids ratio progressively increased with dietary increase of BSFLM in both species. At end of the growth experiment, the levels of catalase, superoxide dismutase and lysozyme increased linearly with the inclusion of BSFLM in both species while malondialdehyde showed similar values between different groups. However, catalase, and superoxide dismutase increased (T0

Investigation on Immune-Related Protein (Heat Shock Proteins and Metallothionein) Gene Expression Changes and Liver Histopathology in Cadmium-Stressed Fish.

Heat shock proteins (HSP) are highly conserved in their structure and released in case of stress. Increased metallothionein (MT) synthesis is associated with increased capacity for binding heavy metals. Healthy juveniles of grass carp were exposed to sublethal dose (1.495 mg L-1) of cadmium for 28 days. Simultaneously, a control group was also run to compare difference of total RNA expression levels in cadmium-treated and control groups. The cadmium levels in the tissues of treated fish recorded were 1.78 ± 0.10 mg L-1, 1.60 ± 0.04 mg L-1, and 2.00 ± 0.05 mg L-1, respectively. Several histological alterations including edema, hemorrhage, dilated sinusoids, hypertrophy, hyperplasia, congestion of central vein, and nuclear alterations were observed in cadmium-exposed fish. Stress gene (metallothionein and heat shock proteins) mRNA transcription levels were studied by mRNA extraction and cDNA preparation by using PCR. The expression level of heat shock protein gene was higher as compared to metallothionein and beta-2-microglobulin gene after cadmium exposure. This study reports various stress-related immune-responsive changes of immune proteins, heat shock proteins, metallothionein, and histopathological changes in fish due to cadmium toxicity that make the fish immunocompromised which may be considered as the biomarkers of cadmium toxicity in other experimental species.

Analysis of the growth performance, stress, profile of fatty acids and amino acids and cortisol in Tilapia (Oreochromis niloticus), cultured at high stocking density using in-pond raceway system.

In-pond raceway system technology (IPRS) was introduced in Pakistan in 2019 as solution for sustainable aquaculture approach by effectively increasing production, reducing pollution and facilitating feed and pond management. Fingerlings of GIFT Tilapia (Oreochromis niloticus) (n = 16,500 in each raceway, initial weight = 32.00 ± 1.26 g) were stocked in June 2019 in two IPRS raceways (area of each raceway = 220 m3) for 171 days until harvested on November 30, 2019. Fingerlings stocked in traditional earthen ponds (area of each pond = 6167 m3) were studied as control (n = 3000 in each pond, initial weight = 32.00 ± 1.26 g). Average harvested biomass from raceways was 57.33 kg/m3 with an average daily weight gain per fish of 4.47 g per day. On the other hand, average harvested biomass from control ponds was 0.38 kg/m3 with an average daily weight gain per fish of 4.60 g per day. Average feed conversion ratio (FCR) in both raceways and control ponds was recorded as 1.25 and 1.24, respectively. Overall survival rate in both raceways and control ponds was above 99%. No sign of any disease was noted at any stage in both study groups. Crude protein and fats contents did not reduce in any raceway despite of high stocking density and sharp seasonal changes. Profile of essential and non-essential amino acids were found to be upto nutritional requirements of adult human. High levels of n-3 and n-6 fatty acids in fish collected from raceways as compared to those in traditional earthen pond show that muscle quality was not compromised due to high stocking density in small area. Return on investment excluding capital cost was 47.05 which implies that IPRS technology can be economically feasible with further modifications.

A Comparative Assessment of Nanotoxicity Induced by Metal (Silver, Nickel) and Metal Oxide (Cobalt, Chromium) Nanoparticles in Labeo rohita.

In the present in vivo study, we provide a comparison of toxicological consequences induced by four different types of spherical nanoparticles (NPs)-silver nanoparticles (AgNPs, 40 ± 6 nm), nickel (NiNPs, 43 ± 6 nm), cobalt oxide (Co3O4NPs, 60 ± 6 nm), and chromium oxide (Cr3O4NPs, 50 ± 5 nm)-on freshwater fish Labeo rohita. Fish were exposed to NPs (25 mg/L) for 21 days. We observed a NPs type-dependent toxicity in fish. An altered behavior showing signs of stress and a substantial reduction in total leukocyte count was noticed in all NP-treated groups. A low total erythrocyte count in all NP-treated fish except for Co3O4NPs was discerned while a low survival rate in the case of Cr3O4NP-treated fish was observed. A significant decrease in growth and hemoglobin were noticed in NiNP- and Cr3O4NP-treated fish. A considerable total protein elevation was detected in NiNP-, Co3O4NP-, and Cr3O4NP-treated groups. An upgrading in albumin level was witnessed in Co3O4NP- and Cr3O4NP-treated groups while a high level of globulin was noted in NiNP- and Co3O4NP-exposed groups. In all NP-treated groups, a depleted activity of antioxidative enzymes and pathological lesions in liver and kidney were noticed.

Evaluation of Fish Health Status and Histopathology in Gills and Liver Due to Metal Contaminated Sediments Exposure.

Health status of freshwater fish, Cirrhina mrigala was studied by qualitative and quantitative histopathological analysis, alterations in frequency/prevalence percentages (%) and histological alteration indices (HAI) in response to metal contaminated sediments of the River Ravi aquatic ecosystem. Histo-structures of gill and liver samples of fish were analyzed and comparison between the degree of damage of the alterations in fish organs was performed after exposure to metal contaminated sediments for 7, 14 and 28 days under semi-static water renewal bioassays. Histopathological studies revealed marked histological alterations in the gills and liver of exposed fish as compared to normal tissue structure observed in control fish. The frequency and prevalence percentages observed in 28-day exposed fish were significantly higher as compared to 7- and 14-day exposed fish. The order of frequency and prevalence percentage for gills and liver of exposed fish was as: 28-day > 14-day > 7-day. The highest prevalence percentages recorded were 83 and 80% as focal area of necrosis in gill and liver, respectively, after 28-day exposure. The lowest prevalence percentage observed in 7-day exposed Cirrhina mrigala was dilation of sinusoids (17%).

Sex reversal of brook trout (Salvelinus fontinalis) by 17α-methyltestosterone exposure: A serial experimental approach to determine optimal timing and delivery regimes.

Commercial culture of Brook trout (Salvelinus fontinalis) in Tasmania was partly abandoned due to sexual maturation of male fish early on during the estuarine rearing phase. Maturation adversely affects body mass, flesh quality and immunocompetency effectively. Sex reversal techniques such as the in-feed addition of a synthetic androgen have proven difficult to adapt in brook trout. An appropriate timing, duration and delivery vehicle for administration of 17α-methyltestosterone (MT) to produce phenotypic males (neomales) from genotypically female brook trout required further investigation. In this study, groups of brook trout eggs (n=1000) maintained at 9.5±0.15-10±0.14°C, were immersed in MT (400µgL-1) for four hours on two alternate days (two immersions/group) staggered over a two week period surrounding the hatch of embryos (control groups excluded). The groups were then split and half received MT-supplemented feed for 60days and the other a standard diet. Following an 11 month on-growing period sex phenotypes were determined by gross & histological gonad morphology. The highest proportion of male phenotypes (75%) was found in fish immersed six and four days pre-hatch and subsequently fed a normal diet. Fish fed a MT supplemented diet and immersed in MT showed significantly higher proportions of sterile fish. These data indicate that a pre-hatch immersion-only regime (4-6days pre-hatch at 9.5°C) should be pursued as a target for optimization studies to further refine the effective concentration and duration of exposure to MT for the successful production of neo-male brook trout.