Immunoadjuvant effect of garlic (Allium sativum)–mineral oil suspension on immunity and resistance to Aeromonas hydrophila infection in rohu,Labeo rohita

The immunomodulatory effect of garlic–mineral oil as a modified adjuvant was investigated in rohu, Labeo rohita along with formalin-killed Aeromonas hydrophila. The garlic extract was not having any proteolytic effect on the tested antigen as marked in SDS-PAGE. The efficacy of graded levels of modified adjuvant formulation was assessed by measuring serum hemolysin and agglutination titers as well as level of protection against A. hydrophila challenge in the immunized fish at days 10 and 30 post-vaccination (dpv). The higher hemolysin and bacterial agglutinin titers along with significantly lower mortality against A. hydrophila challenge in modified adjuvant-based vaccinated fish, particularly at low level of garlic incorporation, indicated the pronounced immunoadjuvant role of this modified adjuvant formulation in rohu.     

A study of the damage of the intestinal mucosa barrier structure and function of <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis> with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

The aim of this study is to explore the effect of Aeromonas hydrophila on the intestinal mucosal barrier structure and intestinal permeability in grass carp (Ctenopharyngodon idella). Histopathological examinations showed that A. hydrophila induced severe intestinal lesions, including inflammatory cell infiltration and intestinal villus fusion and swelling. Messenger RNA (mRNA) expression of the inflammatory cytokines TNF-α, IL-1β, IL-8, IL-10 and MyD88 was significantly increased after infection with A. hydrophila. The permeability of intestinal mucosa was determined using Evans blue (EB) and d-lactic acid. The results indicated that the levels of EB and serum d-lactic acid were significantly increased after infection with A. hydrophila (p < 0.05). Our results also indicated that the intestinal mucosal barrier injury induced by A. hydrophila infection was closely associated with the expression of the tight junction (TJ) protein zonula occludens-1 (ZO-1), occludin, claudin b and claudin c as well as the activity of Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase. Lower mRNA levels of occludin and lower Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activity in the intestines were observed after challenge. ZO-1 and claudin c were significantly increased 24 h after infection with A. hydrophila. The most interesting finding was that claudin b also significantly increased 24 h after challenge and then decreased to lower levels at 72, 120 and 168 h post-infection compared to the PBS-treated control group. The results demonstrated that grass carp infection with A. hydrophila induced intestinal inflammation and impaired the structure and function of the intestinal mucosal barrier.     

Effect of Bacillus subtilis on Aeromonas hydrophila infection resistance in juvenile freshwater prawn, Macrobrachium rosenbergii (de Man)

This experiment was carried out to investigate the potential probiotics properties of Bacillus subtilis in protecting of juvenile freshwater prawn, Macrobrachium rosenbergii against Aeromonas hydrophila infection. A B. subtilis bacterium isolated from gut of juvenile prawns was added to the prawns feed at 10⁸ cells g^?1 feed. There were significant differences (P < 0.05) between B. subtilis-treated and control groups in growth and survival enhancement of juvenile prawns after 60 days of feeding trial. Sixty days after B. subtilis feeding trial, the prawns were challenged by bath exposure to A. hydrophila (10⁷ cells mL^?1) for 28 days. Four weeks after challenge, there was significant difference in the survival of prawns between B. subtilis-treated groups (88.33 %) and control groups (20.81 %; P < 0.05). In addition, the control groups had an unhealthy external appearance, while the treated groups, appeared healthy and normal. From this, it was concluded that the selected B. subtilis may be a promising probiotics for protection of prawns from A. hydrophila infection.     

Antigen encapsulated alginate-coated chitosan microspheres stimulate both innate and adaptive immune responses in fish through oral immunization

The potential of alginate-coated chitosan microspheres to act as an antigen carrier through oral vaccination was explored in fish. Among different antigenic formulations such as antigen-loaded alginate-coated and alginate-uncoated chitosan microspheres, empty alginate-coated chitosan microspheres and free whole-cell antigen, only alginate-coated chitosan microspheres gave promising results for stimulating both adaptive and innate immune response up to 6 weeks of post-immunization. In addition, alginate-coated chitosan microspheres modulated antigen-release behavior, maintained structural integrity and stability and gave effective protection to antigens from acidic degradation in vitro for 12 h. However, none of these formulations tested in this study are effective for protecting fish against Aeromonas hydrophila infection under the conditions evaluated in the present work. The enhancement of the immune response observed with the antigen-loaded alginate-coated chitosan microspheres demonstrates its promising effect for oral antigen delivery in fish.     

Enhancement of the innate immune system and disease‐resistant activity in Cyprinus carpio by oral administration of β‐glucan and whole cell yeast

The effects of dietary β‐ (1,3) glucan and whole cell yeast (Sacharomyces uvarum) on the immune response and disease resistance to Aeromonas hydrophila were investigated in Cyprinus carpio. β‐(1,3) glucan was extracted from the yeast. Both β‐(1,3) glucan and whole yeast were incorporated into the diet at 1% level and fed to common carp C. carpio for a period of 60 days. Control and treated fish were exposed to A. hydrophila on the 30th and the 60th day of the experimental period. Dietary supplementation of glucan significantly increased the white blood cell count in fish on the 60th day (2.91±0.04 × 10⁴), and the highest nuetrophil nitro blue tetrazolium (NBT) activity was also observed in glucan‐fed fish (30th day). A consistent increase in neutrophil (NBT) activity was also observed in whole cell fed fish until the end of the experiment. Similarly, β‐(1,3) glucan and whole cell yeast enhanced the serum lysozyme activity from the 15th day onwards but higher activity was reported on the 30th day in glucan and the 60th day in whole cell yeast‐fed fish. Suplementation of β‐(1,3) glucan protected the fish from A. hydrophila infection. Nearly 75–80% of the fish survived pathogen exposure (relative percentage survival). However, only 54–60% survival was observed in the whole cell‐fed fish. β‐(1,3) glucan and whole cell yeast protect the fish from pathogens by enhancing the cellular and humoral immune response in C. carpio.     

Antibacterial properties of anthraquinones extracted from rhubarb against Aeromonas hydrophila

Antibacterial properties of crude extract from rhubarb and its major bioactive compounds against Aeromonas hydrophila were assayed. Major bioactive compounds (anthraquinone derivatives) in rhubarb collected from different cultivation areas were determined by ultra-performance liquid chromatography (UPLC); the antibacterial activity [minimum inhibitory concentration (MIC)] of rhubarb was positively related to the anthraquinone content (r = 0.9306, P < 0.01). The MIC values of five anthraquinones against A. hydrophila were found to be in the range 50–200 μg/ml. Action-mode studies showed that anthraquinones (emodin) inhibits cellular functions by binding to cell DNA after penetrating the cell membrane, resulting in cell death. The present study suggests that anthraquinones extracted from rhubarb have potential use as antimicrobials for control of A. hydrophila.     

Effect of Coriolus versicolor polysaccharides on the hematological and biochemical parameters and protection against Aeromonas hydrophila in allogynogenetic crucian carp (Carassius auratus gibelio)

The effect of dietary intake of Coriolus versicolor Polysaccharides (CVP) on the hematological and biochemical indices of Allogynogenetic crucian carp (Carassius auratus gibelio) was investigated. Fish were fed CVP supplemented diets (0, 0.25, 0.5, 1.0, 2.0 or 4.0 g CVP kg^?1) for 56 days. The RBC, WBC counts, hemoglobin content, ESR in blood and TP, ALT, AST, ALP, GLU, CHO, TG, and BUN in serum were measured on day 0, 14, 28, 42, and 56. After feeding of 56 days, fish were infected with Aeromonas hydrophila and mortalities were recorded. The results indicated that feeding crucian carp with suitable dose of CVP enhanced the RBC, WBC counts, hemoglobin and TP content, ALP activity, and decreased the ESR, ALT, AST, GLU, CHO, TG and BUN. There was no effect in fish at low dose (0.25 g kg^?1). Unexpectedly, the higher CVP dose used here (2.0 and 4.0 g kg^?1) has a negative effect in fish. The results of challenge experiment indicated that a moderate level of CVP in the diet (1.0 g kg^?1) was the most effective to enhance the survival of fish after infected with A. hydrophila. In summary, the use of CVP, as dietary supplements, can improve the innate defense of crucian carp providing resistance to pathogens.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Expression of genes involved in the early immune response at the distal segment of the gut in Atlantic cod, Gadus morhua L. after vaccination with a bacterial antigen

The gastro-intestinal tract serves as the route of infection of most bacterial pathogens in fish, and the immune response genes in the gut produce substances that provide an initial defense during pathogen invasion. The expression of some genes involved in antibacterial defense, cell-mediated immunity and inflammation in the distal intestine of Atlantic cod, Gadus morhua L., was investigated following intraperitoneal vaccination with heat-killed Vibrio anguillarum. Tissue samples of the distal intestine were collected at 0, 1, 3, 7 and 10 days post-vaccination (dpv) and processed for semi-quantitative RT-PCR analyses. Expression of the bactericidal/permeability-increasing protein/lipopolysaccharide-binding protein (BPI/LBP), g-type lysozyme, non-specific cytotoxic cell receptor protein-1 (NCCRP-1) and interleukin-8 (IL-8) significantly increased during the early stages post-vaccination, that is, 1–3 dpv, followed by a decrease in their expression returning to their initial levels. Transferrin and apolipoprotein A-I, although weakly expressed in the distal intestine, also increased in the vaccinated fish. Interleukin-1β (IL-1β) was significantly decreased in its expression level relative to the PBS-injected fish at 3 and 7 dpv. The expression of these immune-related genes in the distal intestine as a result of vaccination provides valuable information on the mechanisms of gut immunity in the fish against pathogenic bacteria, particularly during the early stages of infection.     

Differences in the antibody response and survival of genetically different varieties of common carp (Cyprinus carpio L.) vaccinated with a commercial Aeromonas salmonicida/A. hydrophila vaccine and challenged with A. hydrophila

Males of two strains of carp, wild Duna (D), and inbred Szarvas 22 (22), were selected for high and low stress response. Two purebreds of D and 22, from randomly chosen parents and four crosses, 22 × 22-L (low stress response), 22 × 22-H (high stress response), 22 × D-L (low stress response) and 22 × D-H (high stress response) from selected stress response parents were produced and vaccinated with a commercial Aeromonas salmonicida/Aeromonas hydrophila vaccine and their circulating antibody response evaluated 1, 3, 5, and 7 weeks post-vaccination by ELISA. Significantly higher titres of circulatory antibodies against A. hydrophila were found in the families 22 and cross 22 × 22-L compared to other groups. The development of circulatory antibodies against A. hydrophila in all crosses having at least one D parent was low and remained low throughout the experiment. The level of circulatory antibodies against atypical A. salmonicida in the inbred strain increased following a booster vaccination with the highest values measured in inbred strain 22 and cross 22 × 22 L. The different varieties of carp had different levels of survival against experimental challenge with A. hydrophila. The greatest survival was obtained in strain 22 and cross 22 × 22-L, while ~90% of D wild carp and cross 22 × D (independent of their stress response) died. Survival results correlated well with the antibody response of the different groups: 22 and 22 × 22-L had the highest antibody titres against A. hydrophila and the greatest level of survival.     

Supplementation of microbial levan in the diet of Cyprinus carpio fry (Linnaeus, 1758) exposed to sublethal toxicity of fipronil: effect on growth and metabolic responses

A 60-day feeding trial was conducted to study the effect of dietary microbial levan on growth performance and metabolic responses of Cyprinus carpio fry exposed to sublethal dose (1/10th LC₅₀) of fipronil [(±)-5-amino-1-(2,6-dichloro-α,α,α-trifluoro-p-tolyl)-4-trifluoromethylsulfinylpyrazole-3-carbonitrile]. Two hundred and twenty five fry were randomly distributed in five treatments in triplicates. Four purified diets were prepared with graded levels of microbial levan. Five different treatment groups were levan control L₀P₀ (basal feed + 0 % levan without exposure to pesticide); pesticide control L₀P₁ (basal feed + 0 % levan with exposure to pesticide); L_0.25P₁ (basal feed + 0.25 % levan with exposure to pesticide); L_0.50P₁ (basal feed + 0.50 % levan with exposure to pesticide); and L_0.75P₁ (basal feed + 0.75 % levan with exposure to pesticide). Weight gain% and specific growth rate were significantly higher (p < 0.05) in levan fed groups compared to their non-levan fed counterpart. Highest (p < 0.05) content of ascorbic acid in muscle, liver and brain tissues was observed with higher level of dietary levan. Glucose-6-phosphate dehydrogenase activity decreased with the increasing level of dietary levan in the liver and muscle. Aspartate aminotransferase activity exhibited a second order polynomial relationship with the dietary levan, both in liver (Y = ?1.001x ² + 5.366x + 5.812, r ² = 0.887) and muscle (Y = ?0.566x ² + 2.833x + 6.506, r ² = 0.858) while alanine aminotransferase activity showed third order polynomial relationship both in liver (Y = 1.195x ³ ? 12.30x ² + 35.23x + 9.874, r ² = 0.879) and muscle (Y = 0.527x ³ ? 8.429x ² + 31.80x + 8.718, r ² = 0.990). Highest (p < 0.05) superoxide dismutase activity in gill was observed in the group fed with 0.75 % levan supplemented diet. Overall results indicated that dietary microbial levan at 0.75 % in C. carpio fry ameliorated the negative effects of fipronil and augmented the growth.     

Hemato-immunological responses of Heros severus fed diets supplemented with different levels of Dunaliella salina

In this study, the effects of oral administration of different levels of Dunaliella salina (a natural β-carotene source) on growth parameters, immunological and hematological indices, as well as skin carotenoids, of Heros severus were investigated. One hundred and eighty H. severus weighing 27 ± 0.5 g were divided randomly into four groups in triplicate (15 fish in each replicate). Groups 1–4 received food supplemented with 0, 50, 100 and 200 mg kg^?1 D. salina powder, respectively. After 6 weeks, the growth parameters were compared among the groups. Blood samples were taken from each group, and hematological parameters including red blood cell count (RBC), white blood cell count (WBC), hematocrit (PCV), hemoglobin (Hb) and immunological indices (serum and mucus lysozyme and bactericidal activity, resistance against Aeromonas hydrophila infection) as well as carotenoid content of skin were evaluated. Results showed that some growth indices increased significantly in fish fed with 100 and 200 mg kg^?1 D. salina-supplemented food (P < 0.05). Although serum lysozyme activity was increased in fish fed with food supplemented with 100 and 200 mg kg^?1 D. salina (P < 0.05), no significant change was observed in serum and mucus bactericidal activity and mucus lysozyme activity among the groups (P > 0.05). Most of the hematological parameters such as WBC, RBC, PCV and Hb significantly increased in D. salina-treated fish compared with controls (P < 0.05). Mortality induced after challenge with A. hydrophila in 200 mg kg^?1 D. salina-treated fish was 36.67 %, which significantly decreased compared with control (P < 0.05). Skin carotenoid content in all D. salina treatments was statistically higher than that of control (P < 0.05). Conclusively, D. salina as a food additive can affect positively the growth, immunological and hematological parameters of H. severus.     

Alteration in certain enzymological parameters of an Indian major carp, Cirrhinus mrigala exposed to short- and long-term exposure of clofibric acid and diclofenac

The extensive use of pharmaceuticals in human and veterinary medicine may enter the aquatic environment and pose a serious threat to non-target aquatic organisms like fish. In this study, Indian major carp Cirrhinus mrigala was exposed to different concentrations (1, 10 and 100 μg L^?l) of most commonly used pharmaceutical drugs clofibric acid (CA) and diclofenac (DCF) to evaluate its impacts on certain enzymological parameters during short- and long-term exposures. During short-term (96 h) exposure period, plasma glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and gill Na⁺/K⁺-ATPase activity were significantly altered at all concentrations of both the CA- and DCF-treated fish. In long-term exposure (35 days), gill Na⁺/K⁺-ATPase activity was found to be significantly increased at all concentration of CA and DCF exposures throughout the study period (except at the end of 7th day in 10 and 100 µg L-1) . However, a biphasic trend was observed in plasma GOT and GPT activity when compared to the control groups. In both short- and long-term exposure, a significant (P < 0.01 and P < 0.05) changes were observed in all enzymological parameters of fish C. mrigala exposed to different concentrations of CA and DCF. The alterations of these enzymological parameters can be effectively used as potential biomarkers in monitoring of pharmaceutical toxicity in aquatic environment and organisms.     

The effects of bioturbation by the Venus clam Cyclina sinensis on the fluxes of nutrients across the sediment–water interface in aquaculture ponds

A mesocosm experiment was conducted to study the effects of bioturbation by the marine bivalve Cyclina sinensis in an aquatic system and to examine how the bioactivities of this clam alter the rate at which nutrients are exchanged across the sediment–water interface. A dark incubation experiment was performed to determine the fluxes of inorganic nutrients (NH₄ ⁺, NO₃ ^? + NO₂ ^? and PO₄ ^3?), benthic chlorophyll a (Chl a), organic matter and sediment oxygen consumption (SOC) on days 1, 10, 20 and 30. C. sinensis destroyed the initial sediment surface and partially enhanced oxygen penetration into the sediment. The clam also increased the water content and volume of the oxic sediment. The analysis showed that increases in the clam density and time period produced higher fluxes of all the nutrients (NH₄ ⁺, NO₃ ^? + NO₂ ^? and PO₄ ^3?) in the bioturbated chambers compared with the control (non-bioturbated chambers). The concentration of Chl a in the sediment increased significantly at the end of the experiment (day 30) (p < 0.05). The SOC rate increased significantly with the increase in density and with the time period from day 10 to day 30 (p < 0.05). As a result, the fluxes of nutrients increased with increased SOC, producing oxygen-induced sediment mineralization. Our findings suggest that the activities of the clams could positively influence SOC and cause an increase in the fluxes of inorganic nutrients. These effects may substantially improve the primary productivity and water quality of earthen pond ecosystems.     

Achyranthes aspera (Prickly chaff flower) leaves- and seeds-supplemented diets regulate growth,innate immunity,and oxidative stress in Aeromonas hydrophila-challenged Labeo rohita

ABSTRACT

The immunostimulatory and disease-resistance properties of Achyranthes aspera were evaluated in rohu (Labeo rohita) challenged with Aeromonas hydrophila. Experimental diets were enriched with leaves at 0.25% (D1) and 0.5% (D2) and seeds at 0.5% (D3); the control diet (D4) was without any enrichment. Rohu (2.02 ± 0.23 g) were cultured for 75 days and then challenged with bacteria. The highest average weight was observed in the D3 diet-fed fish. The cumulative mortality rates were 70%, 60%, 40%, and 30% in the D4, D1, D2, and D3 diets fed to rohu respectively. Enriched diets significantly increased myeloperoxidase, nitric oxide synthase, and serum lysozyme levels and decreased malondialdehyde and carbonyl protein content. Expressions of lysozyme C and lysozyme G were significantly (P < .05) higher in the D3 diet-fed fish. In the kidney, IL-1β and TLR 4 were up-regulated in enriched-diet-fed fish. Supplementation of seeds and leaves at 0.5% showed a positive impact in fish.     

Development of a simple and rapid monoclonal antibody-based flow through immunogold assay (FIA) for detection of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

In this study, we developed a simple, low-cost, and rapid flow through immunogold assay (FIA) for detection of Aeromonas hydrophila in fish tissues and validated. The developed assay relied on colloidal gold conjugated highly specific monoclonal antibody (B8E3) against 20 kDa protein of A. hydrophila. The assay can be completed within 5 min including antigen (sample) preparation and exhibited no cross-reaction with other major aquatic pathogens such as Vibrio anguillarum, V. parahaemolyticus, V. harveyi, white spot syndrome virus (WSSV), and Aphanomyces invadans. A wider range of Aeromonas species were tested including A. caviae, A. dhakensis, A. sobria, A. veronii, A. culicicola, A. sharmana, and A. schubertii and no cross-reactivity could be observed. The developed technique FIA could detect 10³ CFU/ml of aeromonad cells. In addition, the FIA is as sensitive as the first-step polymerase chain reaction (PCR) and 1000 times sensitive than the immunodot blot method. The validation study revealed the accuracy of FIA by comparing with the standard first-step PCR. The proposed FIA is low-cost, easy-to-use, and compatible for field-level analysis of A. hydrophila in fish tissues. Our developed FIA could successfully detect A. hydrophila at early stage of the infection and could be easily availed by farmers, fish producers, non-technicians, and technicians in the aquaculture industry.     

Improvement of nutritive value of sesame oilseed meal in formulated diets for rohu,Labeo rohita (Hamilton), fingerlings after fermentation with two phytase-producing bacterial strains isolated from fish gut

Eight isonitrogenous (approximately 35 % crude protein) and isocaloric (17.58 kJ g^?1) diets incorporating raw and fermented sesame oilseed meal replacing other feed ingredients including fishmeal at 10, 20, 30 and 40 % levels by weight into a fishmeal-based reference diet (RD) were fed to rohu, Labeo rohita, fingerlings (mean initial weight 3.19 ± 0.09 g) for 80 days. Two phytase-producing bacterial strains (LF1 and LH1 of Bacillus licheniformis) isolated from the foregut and hindgut regions of adult L. rohita were used for fermentation of oilseed meal for 15 days at 37 ± 2 °C. Fermentation of sesame seed meal was effective in significantly reducing the crude fibre content and anti-nutritional factors such as tannins and phytic acid and enhancing available free amino acids, free fatty acids and mineral concentration. In terms of growth, feed conversion ratio and protein efficiency ratio, 30 % fermented oilseed meal incorporated diet resulted in a significantly (P < 0.05) better performance of rohu fingerlings. In general, growth and feed utilization efficiencies of diets containing fermented oilseed meal were superior to diets containing raw meal. The apparent digestibility of protein, lipid, ash and minerals (phosphorus, calcium, manganese, copper and iron) was significantly (P < 0.05) higher in fish fed diet containing 30 % fermented oilseed meal in comparison with those fed RD. The maximum deposition of protein in the carcass was recorded in fish fed the diet containing 30 % fermented seed meal.     

Immune effects of bathing European eels in live pathogenic bacteria,Aeromonas hydrophila

The specific and non‐specific immune parameters and protection of European eels (Anguilla anguilla) were evaluated after bathing eels with Aeromonas hydrophila. Two hundred eels were distributed into two equal groups and bathed with Phosphate‐buffered saline (Control group) or 1.0 × 10⁷ cfu mL^?1 A. hydrophila (Test group) for 1 h respectively. Then, eels were bled aseptically from the caudal sinus on 1, 4, 7, 14 and 28 days post treatment. The blood cells were used to evaluate the cellular immunity and the serum was used to determine the titres of specific antibody as well as the activities of superoxide dismutase (SOD) and lysozyme. Eels from both groups were challenged by intraperitoneal injection of 1.0 × 10⁷ cfu of A. hydrophila on 28 and 42 days post bathing. The results show that eels bathed in A. hydrophila significantly (P < 0.05) enhanced the proliferation of different types of blood cells and the serum titres of anti‐A. hydrophila antibody. The Relative Percent Survival (RPS) after challenge on 28 and 42 days post bathing in Test group vs. Control group was 40% and 50% respectively. These results suggest that bathing European eels in A. hydrophila would positively affect specific as well as non‐specific immune parameters and protect against A. hydrophila infection in freshwater farming.     

Substitution of soybean meal with fermented <Emphasis Type="Italic">Jatropha</Emphasis> kernel meal: effect on growth performance,body composition,and metabolic enzyme activity of <Emphasis Type="Italic">Labeo rohita</Emphasis>

A 60-day feeding trial was conducted to evaluate the nutritional potential of fermented Jatropha kernel meal (FJKM) in the diets of rohu, Labeo rohita, fingerlings. Four iso-nitrogenous (336 g kg^?1) and iso-energetic (20 MJ kg^?1) diets containing 0, 100, 200, and 300 g kg^?1 FJKM in replacement of 0, 33.3, 66.7, and 100% soybean meal protein, respectively, were prepared and designated as control, T1, T2, and T3, respectively. A total of 180 acclimatized fish (average weight 6.05?±?0.04 g) were equally distributed into four experimental groups in triplicates, and were fed their respective diets to satiation twice daily at 10:00 h and 18:00 h. Higher percent weight gain (%WG); specific growth rate (SGR); protein efficiency ratio (PER); protease, aspartate amino transferase (AST), and alanine amino transferase (ALT) activities; and lower feed conversion ratio (FCR) were recorded in T2 and T3 groups compared to other groups. Whereas hepatosomatic index (HSI), intestinal somatic index (ISI), and amylase, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and catalase activities among the various dietary groups did not vary significantly (P?>?0.05). Whole body composition of fish analyzed at the end of the feeding trial exhibited significantly (P?<?0.05) higher ether extract and lower crude protein in the control group compared to the FJKM-fed groups. These results indicated that rohu fingerlings can efficiently utilize FJKM without any detrimental effect on growth performance, nutrient utilization, and metabolic response.     

Effect of Ocimum sanctum Linn. (Tulsi) extract on the immunity and survival of Labeo rohita (Hamilton) infected with Aeromonas hydrophila

A 60 days study was conducted to evaluate the efficacy of water extract of Ocimum sanctum Linn. leaf on the immune response and disease resistance of Labeo rohita fingerlings against the Aeromonas hydrophila infection. Ocimum sanctum extract was incorporated in the diets (at 0.0%, 0.05%, 0.1%, 0.2%, 0.5% and 1%) of Labeo rohita, rohu fingerlings (6.6 ± 0.013 g). After 42 days blood, plasma and serum were sampled to determine super oxide anion production, lysozyme activity, total immunoglobulin in plasma, blood glucose, serum total protein, albumin, globulin, albumin:globulin ratio, WBC, RBC, haemoglobin content. Fish were challenged with A. hydrophila after 42 days and mortalities were recorded over 18 days post infection. The results demonstrate enhanced super oxide anion production, lysozyme activity, total immunoglobulin in plasma, serum total protein, globulin, total RBC counts, total WBC counts and haemoglobin content (P < 0.05) in treatments group compared with control group. Dietary O. sanctum extracts of 0.2% showed significantly (P < 0.05) higher protection relative percentage survival (RPS 40.00 ± 5.773%) against A. hydrophila infection than control. These results indicate that O. sanctum leaf extract stimulates the immunity and makes L. rohita more resistant to bacterial infection (A. hydrophila).