First record of Cymothoa indica (Crustacea, Isopoda, Cymothoidae) infecting the cultured catfish Mystus gulio in India

Cymothoa indica (Isopoda, Cymothoidae) is reported parasitizing the long-whiskered catfish Mystus gulio, cultured in an experimental cage in India. The specimens observed were adult males and females, which had mainly settled in the buccal cavity of juvenile catfish. The species was previously known from wild populations of Siganidae and Belonidae, but this is the first record of C. indica parasitizing the cultured long-whiskered catfish. Serious lesions, typical of a crustacean infection, were macroscopically visible inside the buccal cavity. The cumulative mortality, over a period of 10 d, was 100%. The parasitic problem was not successfully dealt with, due to an unexpected prevalence and very swift mortality.     

Dietary immunostimulants influence specific immune response and resistance of healthy and immunocompromised Asian catfish Clarias batrachus to Aeromonas hydrophila infection

In order to determine the efficacy and immunoreversal effect of the 4 dietary immunomodulators, viz. lactoferrin, beta-1,3 glucan, levamisole and vitamin C, on disease resistance of a commercially important catfish, Clarias batrachus, fish were fed diets supplemented with various levels of these substances in 2 subgroups, healthy and immunocompromised, during a 30 d trial. An artificial immunosuppressive state was induced by giving 3 intraperitoneal (i.p.) injections of cyclophosphamide (CYP) at a dose level of 200 mg kg(-1) body weight at 1 wk intervals in the immunocompromised vaccinated subgroup and 3 consecutive injections 3 d before challenge in the immunocompromised non-vaccinated subgroup. On the first day of the experiment, the fish were vaccinated against a formalin-killed Aeromonas hydrophila bacterin. After 30 d, antibody titre (as measured through bacterial agglutination titre) and disease resistance against A. hydrophila were determined. The results demonstrate that all 4 immunomodulators were capable of significantly (p < 0.05) enhancing the specific immune response; this was evident through raised antibody titre and protection against A. hydrophila in both healthy and immunocompromised vaccinated subgroups compared to their respective controls. Similarly, all 4 substances significantly raised the survival rates in immunocompromised and healthy non-vaccinated fish. Thus, these substances were capable of reducing the immunosuppression induced by CYP injections in both vaccinated and non-vaccinated fish compared to their respective controls. Among the 4 substances studied, beta-1,3 glucan was found to be the most effective immunomodulator, followed by levamisole, lactoferrin and vitamin C in Asian catfish. Therefore, the results support the introduction of these substances into the diet of fish grown in farms under immunosuppressive/stressful conditions in order to enhance protection against infection and offer economic benefits.     

The effect of incubation temperature, sodium chloride and ascorbic acid on the growth kinetics of Aeromonas hydrophila

The effects of temperature, sodium chloride and ascorbic acid on the aerobic growth kinetics of a clinical strain of Aeromonas hydrophila were evaluated. At 5°C, ascorbic acid (1 mmol l^-1) and sodium chloride (3% w/v) inhibited the growth of the organism. At 10°C, ascorbic acid depressed only the maximum population densities (A) by approximately 2 log cycles, but not maximum specific growth rate (μ_m) or the lag time (Λ). On the contrary, NaCl caused A to increase, with the effect being greatest when the NaCl content was 1.5%. Temperature increase from 10 to 15°C resulted in an approximate doubling of μ_m and unexpectedly an apparent increase in Λ However, this apparent increase resulted from the particular manner in which the lag phase was mathematically calculated.     

Preponderance of GC-rich sites in silver-stained nucleolus organizing regions of Rita rita (Hamilton) and Mystus gulio (Hamilton) (Bagridae, Pisces), as revealed by chromomycin A3-staining technique and scanning electron microscopic studies

The karyotypes of two species of catfish, Rita rita (Hamilton) (2n = 54; 14m + 34sm + 6st; NF = 102) and Mystus gulio (Hamilton) (2n = 58; 30m + 12sm + 2st + 14t, NF = 100) were studied through Giemsa-, silver- and chromomycin A(3)-staining techniques. The silver-stained karyotypes in both sexes of R. rita and M. gulio revealed that the nucleolus organizing regions were located terminally at the shorter arms (Tp) of one pair of submetacentric chromosomes, placed at positions Nos. 2 and 1, respectively, which was confirmed by scanning electron microscopy. Staining with a GC-specific fluorochrome, chromomycin A(3), produced bright fluorescence in the Ag-positive nucleolus organizer regions, suggesting thereby that nucleolus organizing regions actually included GC-rich sites of active r-RNA genes in metaphase chromosomes of these two bagrids. Further such studies are needed due to the extreme paucity of data on fish.     

Dietary ascorbic acid may be necessary for enhancing the immune response in Siberian sturgeon (Acipenser baerii), a species capable of ascorbic acid biosynthesis

Siberian sturgeon (Acipenser baerii Brandt) is one of few fish species capable of synthesizing ascorbic acid (AA); however, it is unknown whether dietary AA is still necessary for this species to generate an optimal non-specific immune response, as is found in most teleostean species that do not synthesize AA. Sturgeon fed either 0 or 500 mg AA (AA polyphosphate) equivalent kg(-1) diets for 2 weeks were then either sham or experimentally injected with Aeromonas hydrophila lipopolysaccharide (LPS) to mimic infection and therefore instigate a non-specific immune response. No significant differences were found in growth and survival throughout the duration of the experiment between sham and LPS groups and between both diets. Two and eight days after injection, serum lysozyme levels were elevated in both LPS and non-LPS groups. Respiratory burst was not affected by dietary AA and significantly higher in LPS groups over non-LPS groups; however, dietary AA may have enhanced the effects seen in the LPS injected fish fed the AA supplemented diet. There were no significant differences in renal l-gulono-1, 4-lactone oxidase (GLO) activity between dietary and LPS treatments, but fish exposed to LPS without dietary AA demonstrated an inter-organ transfer of AA from posterior kidney to liver. Our results indicate that dietary AA may be conditionally necessary for Siberian sturgeon to achieve optimal immune response, particularly in early developmental stages, information imperative to developing successful aquaculture programs for sturgeon species.     

Effect of Aeromonas proteases on the binding of Aeromonas hydrophila strains to connective tissue proteins

125I-labelled connective tissue protein binding to cells of Aeromonas hydrophila, A. caviae, and A. sobria strains isolated from diseased fish, was correlated with the Aeromonas protease degradation of 125I-labelled collagen types I and IV, fibronectin and laminin, immobilized on tissue culture microtitre plates. An inverse relation between 125I-labelled connective tissue protein binding to cells of Aeromonas strains and proteolytic degradation of immobilized connective tissue proteins by Aeromonas proteases was established. Inhibition of the Aeromonas proteolytic activity by protease inhibitors enhances the 125I-labelled connective tissue protein binding to cells of Aeromonas hydrophila strains. Culture conditions were found to influence both expression of proteolytic activity and binding properties.     

The identity and distribution of striped bagrid catfish,Mystus tengara (Hamilton 1822) revealed through integrative taxonomy

Molecular Biology Reports - The taxonomic status and geographical distribution of M. tengara are vague. No genetic diversity and phylogenetic study have been done till now to resolve its identity...     

Feeding ecology of a bagrid catfish,Mystus tengara (Hamilton, 1822) in the Tanore wetland of Rajshahi,Northwestern Bangladesh

The feeding ecology of Mystus tengara (Hamilton, 1822) was studied in the Tanore wetland of Rajshahi, northwestern Bangladesh from January 2011 to December 2011. In fish purchased from the local market, a total of 14 713 food items were identified in the stomach contents of 3191 M. tengara specimens, an omnivorous catfish with a diversified feeding strategy including cladocerans, copepods, rotifers, gastropods, green algae, diatoms, insects, ‘small fish remains, fish fry and eggs’ and ‘other’ food items. Insects were the most dominant group of prey throughout the year. Seasonal changes in diet composition were due to the availability of prey items and fish size. Ontogenetic changes were noted between the juveniles and adults. The gut contents of M. tengara showed that the food items were from different layers of the water bodies.     

Effect of long photoperiod on the maturation of ovary of the catfish, Mystus tengara (Ham.)

A study has been made of the effect of photoperiodism on the ovarian maturation of fresh water teleost, Mystus tengara (Ham.). The magnitude of ovarian recrudescence is more during maturing virgin phase as compared to during immature virgin and preparatory virgin phases in long photoperiod (14 hours/day) treated fish. Thus, the response of the ovary to long photoperiod increases with the approach of natural spawning period. The mature eggs are more in long photoperiod treated fish than in controls exposed to 12 hours/day photoperiod. In long photoperiod treated fish, prespawning period is reduced considerably. During spawning and post-spawning periods, the ovaries are refractory, since long photoperiod is ineffective in maintaining gravid ovaries beyond spawning period and the ovarian regression during post spawning period sets in regardless of nature of photoperiod treatment.     

Observations on the structure and histochemistry of the skin of Mystus gulio (Ham.) (Osteichthyes: Siluriformes, Bagridae)

The skin of Mystus gulio (Ham.) is typical of siluroids in not having any scales. Large number of mucous cells and sacciform granular cells, interspersed with taste buds, were found in the epidermis. Histochemical tests showed the presence of weakly sulphated mucopolysaccharides in the mucous cells, while the sacciform granular club cells were found to be proteinaceous, with a small amount of lipids and phospholipids. The dermis is composed of a stratum laxum and a stratum compactum. The subcutis was found to consist of loosely arranged collagen fibres and fat cells.     

Modulation of the immune response to an Aeromonas hydrophila aroA live vaccine in rainbow trout: effect of culture media on the humoral immune response and complement consumption

The Aeromonas hydrophila aroA is an attenuated strain that has been assessed as a live vaccine in rainbow trout, Oncorhynchus mykiss. In this study the effects of different culture media used to grow the strain on its survival after in vitro exposure to rainbow trout serum, and on its immunogenicity in rainbow trout were compared. Four culture media were tested: Luria broth (LB), Luria broth with 0.25% glucose, trypticase soy broth (TSB), and brain-heart infusion broth (BHIB). Bacteria grown in culture media with glucose (TSB, BHIB and LB with 0.25% glucose) showed reduced complement consumption and a lower serum susceptibility. O. mykiss vaccinated with inocula prepared with BHIB- and LB-grown aroA cells resuspended in phosphate-buffered saline (PBS) showed higher and longer-lasting serum agglutinating antibody titres than those vaccinated with TSB-grown bacteria. Thus, a direct relationship between serum resistance and immunogenicity could not be established, but BHIB and LB culture media were the most effective in increasing the immunogenicity of the A. hydrophila aroA vaccine.     

Coriolus versicolor polysaccharides enhance the immune response of crucian carp (Corassius auratus gibelio) and protect against Aeromonas hydrophila

To investigate the effect of Coriolus versicolor polysaccharides (CVPS) on the immune response of crucian carp (Corassius auratus gibelio), fish were fed diets containing CVPS at different concentrations including 0.5, 1.0 and 2.0 g kg^?1 for 5 weeks. Other groups of fish were vaccinated by intraperitoneal injection (i.p.) against Aeromonas hydrophila with a killed bacterin at the beginning of the experiment and fed the same diets as described above. Additionally, control fish and vaccinated‐only fish were fed basal diets without CVPS supplementation. The phagocytosis, lysozyme activity, complement components C3 and C4, E‐C3bRR (Erythrocyte‐C3b rosette rate) and E‐CIRR (Erythrocyte‐immune complex rosette rate) levels and circulating antibody titers in the serum were monitored. Five weeks after feeding the prescribed diet, fish were challenged with A. hydrophila and the mortalities recorded. Results showed that feeding non‐vaccinated and vaccinated crucian carp with CVPS stimulated the phagocytosis of leukocytes, lysozyme, complement components C3 and C4, erythrocyte immune adherence, and circulatory antibody titers in serum in vaccinated crucian carp. Immune parameters increased to their highest levels after 3 weeks of feeding the diet containing 0.5 or 1.0 g kg^?1 CVPS. These doses also resulted in the highest protection in the challenge experiment. Best survival (85%) was in the vaccinated group fed the diet containing 1.0 g kg^?1 CVPS, whereas almost 80% of control fish (negative control) and 50% of vaccinated‐only fish (positive control) died.     

Chinese herbs (Astragalus radix and Ganoderma lucidum) enhance immune response of carp, Cyprinus carpio, and protection against Aeromonas hydrophila

The effect of Chinese herbs (Astragalus radix and Ganoderma lucidum) on immune response of carp was investigated. Fish were fed diets containing Astragalus (0.5%), Ganoderma (0.5%) and combination of two herbs (Astragalus 0.5% and Ganoderma 0.5%) for 5 weeks. Other groups of fish were vaccinated (i.p.) against Aeromonas hydrophila/Aeromonas salmonicida (Shering Plough, Essex, U.K.) at the beginning of the experiment and fed the same diets as described above. Control fish (negative control) and fish vaccinated only (positive control) were fed basal diets without supplements of herbs. The respiratory burst activity, phagocytosis, lysozyme activity and circulatory antibody titres in plasma were monitored. Following 5 weeks after feeding, fish were infected with A. hydrophila and mortalities were recorded. The results of this study showed that feeding non-vaccinated and vaccinated carp with combination of Astragalus and Ganoderma stimulated respiratory burst activity, phagocytosis of phagocytic cells in blood and lysozyme and circulatory antibody titres in plasma in vaccinated carp. Fish challenged with A. hydrophila had variable survival. The best survival (60%) was in vaccinated group fed with both herbs, while almost 90% of control fish (negative control) and 60% of fish vaccinated only (positive control) died.     

Effects of dietary cholesterol on antioxidant capacity,non-specific immune response,and resistance to Aeromonas hydrophila in rainbow trout (Oncorhynchus mykiss) fed soybean meal-based diets

This study evaluated the effects of dietary cholesterol on antioxidant capacity, non-specific immune response and resistance to Aeromonas hydrophila in rainbow trout (Oncorhynchus mykiss) fed soybean meal-based diets. Fish were fed diets supplemented with graded cholesterol levels (0 [control], 0.3, 0.6, 0.9, 1.2, and 1.5%) for nine weeks. The fish were then challenged by A. hydrophila and their survival rate recorded for the next week. Dietary cholesterol supplementation generally increased the serum and hepatic superoxide dismutase (SOD), glutathione-peroxidase (GSH-Px), catalase (CAT), and total antioxidant capacity (TAC) activities, but decreased the serum and hepatic malondialdehyde (MDA) contents. Further, the hepatic CAT and serum SOD, CAT, and TAC activities were significantly higher in fish fed diets supplemented with 0.9 or 1.2% cholesterol compared to those fed the control diet, whereas the serum and hepatic MDA contents were significantly lower. The respiratory burst activity, alternative complement activity, and hepatic lysozyme activity increased steadily when the supplemental cholesterol was increased by up to 1.2% and then declined with further addition. The serum lysozyme activity and phagocytic activity increased steadily with increasing dietary supplemental cholesterol level up to 0.9% and then declined with further addition. Dietary cholesterol supplementation generally enhanced the protection against A. hydrophila infection, and fish fed diets supplemented with 0.9 or 1.2% cholesterol exhibited the highest post-challenge survival rate. The results indicated that cholesterol may be under-supplied in rainbow trout fed soybean meal-based diets, and dietary cholesterol supplementation (0.9–1.2%) contributed to improved immune response and disease resistance of rainbow trout against A. hydrophila.     

Effect of ascorbic acid on thioglucosidases from different crucifers

Thioglucosidase activity was demonstrated in partially-purified preparations from several Cruciferae oilseeds, both in the presence and absence of ascorbic acid. The amount of activation by ascorbic acid differed among the enzyme preparations from different species. Buffer composition and pH were found to significantly affect enzyme activity, the turret rape enzyme showing a second optimum at pH 7·1 in the presence of ascorbic acid and sodium phosphate buffer. Disc electrophoresis on polyacrylamide gel revealed distinct isoenzyme patterns from crude extracts of all nine species or varieties studied. Some differences in the patterns were noted from electrophoresis of partially-purified preparations. Ascorbic acid was found to affect isoenzyme patterns and the rate of development of equivalent isoenzymes from yellow mustard and from turret rape.     

Effect of orally administered azadirachtin on non-specific immune parameters of goldfish Carassius auratus (Linn. 1758) and resistance against Aeromonas hydrophila

Modulation of the immune responses using active bio-ingredients as a possible prophylaxis measure has been novel prospect for aquaculture. The present study evaluated the effects of azadirachtin EC 25% on non-specific immune responses in goldfish Carassius auratus and resistance against pathogenic bacteria Aeromonas hydrophila. The experimental trial for effects of azadirachtin on immuno-haematoloical parameters in goldfish was conducted by feeding the various levels of azadirachtin as control T₀ (without azadirachtin), T₁ (0.1%), T₂ (0.2%), T₃ (0.4%), T₄ (0.8%) and T₅ (1.6%) for a period of 28 days. Fishes were challenged with A. hydrophila 28 days post feeding and relative percentage survival (%) was recorded over 14 days post infection. Immuno-haematoloical (total erythrocyte count, total leukocyte count, haemoglobin, packed cell volume, NBT activity, phagocytic activity, serum lysozyme activity, myeloperoxidase activity, total immunoglobulin) and serum biochemical parameters (serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and blood glucose) of fishes were examined at 14 and 28 days of feedings. Fish fed with azadirachtin, showed significantly (p < 0.05) enhanced TEC, TLC, Total Ig, total protein, NBT activity, serum lysozyme activity and myeloperoxidase level in different treatment groups in comparison with control group. Similarly, SGOT, SGPT and blood glucose level were found to be significantly (p < 0.05) high but PCV and Hb did not differ significantly (p > 0.05) in the treatment groups compared to control groups. Azadirachtin at the concentration of 4 g kg^?1 showed significantly (p < 0.05) higher relative percentage survival (42.60%) when compared with the control against A. hydrophila infection. This study indicated that azadirachtin EC 25% (4 g kg^?1) showed higher NBT activity, serum lysozyme, protein profiles, leukocyte counts and resistance against A. hydrophila infection and thus, can be used as a potential immunostimulant in aquaculture.     

Effect of pre-incubation temperature on the lag time of Aeromonas hydrophila

The effect of prior incubation temperature (5, 15, 25 and 35°C) on the lag times of two strains of Aeromonas hydrophila at the same four incubation temperatures was assessed. When grown at 5°C the type strain (ATCC 7966) showed a 28-fold difference in lag time depending on whether the inoculum had been incubated at 5 or 35°C. However, the corresponding factor for a food-derived strain was only 4. A similar effect was observed, to a lesser extent, under most of the conditions used. Prior incubation temperature had no effect on generation time.     

Beta2-microglobulin is involved in the immune response of large yellow croaker to Aeromonas hydrophila: A proteomic based study

Outbreaks of infectious diseases in cultured large yellow croaker have resulted in great economic losses. However, information regarding its immune defense is limited. In the present study, an approach by the combination of differential proteomics with EST resource was applied for investigation of a profile of serum immune response by large yellow croaker to Aeromonas hydrophila challenge after immunization and for characterizing of one of the targeted immune molecules. Of the twelve altered proteins involved in the response, eight were identified by MS, in which three were randomly selected for antiserum preparation and were further confirmed by Western blotting. Furthermore, three β₂m clones, one of the altered molecules, were obtained from a previously constructed Kidney Smart cDNA library of this fish, and were compared for their identity, which contributed to the identification of β₂m cDNA diversity. Meanwhile, the up-regulated β₂m in response to the bacterial immunization and challenge was further confirmed by Western blotting. Our results indicate that β₂m is involved in the immune response of large yellow croaker to the challenge by A. hydrophila after immunization, which suggests an efficient approach for characterizing of targeted molecules at both the gene and protein levels.