Assessment the efficacy of some various treatment methods, in vitro and in vivo, against Aeromonas hydrophila infection in fish with regard to side effects and residues.

Aeromonas hydrophila is an opportunistic bacteria with an overwhelming impact on fish farming industry especially with upraising of drug resistant mutants. This study aimed to evaluate and compare the therapeutic and side effects of levofloxacin (LEV), chitosan-nanoparticles (CNPs), and fructooligosaccharides (FOS) in control of this infection in tilapia. A total of 160 Nile-tilapia divided into 8-groups; G1: negative-control, G2: infected-control, G3: non-infected-(levofloxacin (LEV) 10 mg/kg bwt), G4: non-infected-(chitosan-nanoparticles (CNPs) 1 g/kg ration), G5: non-infected-(fructooligosaccharides (FOS) 20 g/kg ration), G6: infected-LEV, G7: infected-CNPs and G8: infected-FOS for 7 days. MICs were (0.125 µg/ml and 1.25 mg/ml) for LEV and CNPs respectively. No mortalities or significant adverse effects were recorded in non-infected treated-groups while infected were (20%) LEV, (30%) CNPs, (40%) FOS and (70%) G2. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) decreased by LEV and CNPs than FOS while all increased total protein (TP) and albumin than G2. Malondialdehyde (MDA) significantly decreased and superoxide dismutase (SOD) and reduced glutathione (GSH) increased in all infected-treated groups than G2 in various degrees. Urea and creatinine descending order were FOS, LEV then CNPs decreased significantly than G2. LEV musculature residues, using HPLC, decreased gradually till the 5th day; 621.00 ± 0.66, 270.00 ± 0.48 then 64.00 ± 0.40, and 471.00 ± 0.79, 175.00 ± 0.52 ppb then not detected at 1st, 3rd, and 5th days of withdrawal in non-infected and infected groups respectively. Finally, LEV and CNPs were superior as bactericidal, decreasing mortalities and enzyme activities while CNPs and FOS increased performance, non-specific immunity, and antioxidant biomarkers.

Protective Effect of Leek Extract (Allium ampeloprasum L.) on Catfish (Clarias gariepinus) Experimentally Challenged with Aeromonas hydrophila.

BACKGROUND AND OBJECTIVE: Leek (Allium ampeloprasum) is one of the most commonly used herbal foods all over the world. This study was conducted to evaluate the protective effect of leek extract on catfish experimentally challenged with Aeromonas hydrophila, a problematic bacterial pathogen that affects various freshwater fish species. MATERIALS AND METHODS: Aeromonas hydrophila was isolated and identified from catfish showing clinical signs of septicemia. The in vitro activity of leek extract to control the growth of Aeromonas hydrophila was investigated. In the in vivo experiment, about 240 adult catfish (Clarias gariepinus) were fed three different leek extract concentrations (10, 25 and 50 mg kg-1 body weight) for 1 month. Later on, a challenge study was conducted using an identified A. hydrophila strain. Morbidity and mortality were recorded throughout one week post-challenge. Furthermore, the effect of leek extract on some immune-related genes was investigated. RESULTS: Under the in vitro testing, a significant increase (10 and 13 mm) in the inhibition zone was recorded in wells treated with 25 and 50 mg L-1 leak extract, respectively. A significant reduction in fish mortalities was reported in all leek extract treated groups compared to the control group which was given water. TLR1 gene expression was upregulated in fish treated with leek extract while TNFα gene expression was down-regulated. CONCLUSION: Overall, results suggested that the leek extract has immunostimulating effects that can help control bacterial infections in catfish and probably other fish species.

Dual effect of selenium loaded chitosan nanoparticles on growth, antioxidant, immune related genes expression, transcriptomics modulation of caspase 1, cytochrome P450 and heat shock protein and Aeromonas hydrophila resistance of Nile Tilapia (Oreochromis niloticus).

Nowadays there is a great attention for nanotechnology in aquaculture production. It has an efficient role in nutrients and drugs delivery, ponds sterilization, water treatment and aquatic diseases reduction. Till now, there is no available data on impact of selenite-loaded chitosan nanoparticles (SeChNPs) on Nile tilapia. Hence, the current study investigated the effects of selenite-loaded chitosan nanoparticles supplementation on the growth, immune, antioxidant and apoptotic related genes as well as resistance to Aeromonas hydrophila of Nile tilapia, Oreochromis niloticus. A total of 400 fish were randomly divided into four groups, and each group retained five replicates. The control group was fed a basal diet (with inorganic se), other groups fed diets supplemented with SeChNPs 0.5, 1 and 2 g/kg diet. The loading concentration of Se to ChNPs was 0.3, 0.6 and 1.2 mg/0.5, 1 and 2 gm respectively. Fish groups fed SeChNPs (0.5 and 1 g/kg) exhibited the highest final body gain, better feed utilization. Additionally, the expression of myostatin gene was down-regulated by 0.2 and 0.3 fold in group fed 0.5 and 1 g/kg SeChNPs when compared with control group. Dietary inclusion of SeChNPs increased serum lysozyme, alternative complement and myeloperoxidase activities and immunoglobulin type M level. Supplementation of SeChNPs at the level of 2 g/kg up-regulated glutathione peroxidase, superoxide dismutase and catalase expression by 1.12, 4.9 and 2.31 folds respectively, in comparison with control group. In contrast, the levels of C- reactive protein and malondialdehyde were reduced. The expression of IL-10, IL-8, TNF-α and IL-1ß genes was up-regulated after dietary inclusion of different levels of SeChNPs in a dose dependent manner. Post-challenge, the highest survival rate was detected in group fed 2 g/kg SeChNPs (93%) in contrast, the control group was displayed the lowest survival rate (45%). After challenge with A. hydrophila, the expression of caspase 1 was up-regulated in groups fed 1 and 2 g/kg of SeChNPs. Moreover, the maximum down-regulation of cytochromes P450 and heat shock protein were found in 2 g/kg SeChNPs supplemented group (reduced by 0.4 and 0.6-fold, respectively, when compared with control group). In conclusion, the ameliorative effects of SeChNPs on Nile tilapia growth resulted from immune stimulatory and free radicals scavenging effects of selenium loaded chitosan nano composite.

Rheum officinale extract promotes the innate immunity of orange-spotted grouper (Epinephelus coioides) and exerts strong bactericidal activity against six aquatic pathogens.

We investigated the antimicrobial properties and the effects of Rheum officinale extract (ROE) on nonspecific immune parameters of orange-spotted grouper (Epinephelus coioides) in vitro and in vivo. The in vitro analysis was conducted by treating grouper primary head kidney leukocytes with various concentrations of ROE. The phagocytic rate of the leukocytes was elevated in a dose-dependent manner from 0.01 to 0.1 mg/ml, but decreased with higher concentrations of ROE (0.5 and 1.0 mg/ml). The production of reactive oxygen species (ROS) was strongly enhanced in a dose-dependent manner by treatment with ROE doses of 0.1-10.0 mg/ml. However, morphological changes (e.g., rounding and shrinkage of cells, chromatin condensation, fragmentation, and appearance of apoptotic bodies) were observed in the leukocytes after incubation with higher concentrations of ROE (1.0 and 10.0 mg/ml). A 28-day feeding trial was performed to assess the impact of dietary administration of ROE on grouper innate immunity parameters. Fish were fed with feed supplemented with 0, 0.1, 1.0, or 5.0 g ROE per kg of feed. The phagocytic activity of the animals' leukocytes was significantly elevated in all ROE-fed groups on day 1 and in groups fed with ROE at 0.1 or 1.0 g/kg on day 14. Production of ROS was substantially increased on day 1 in fish fed with ROE at 1.0 and 5.0 g/kg, but decreased steadily later on. The ability to generate ROS increased steadily until day 7 in fish fed the lowest concentration of ROE (0.1 mg/ml), but decreased thereafter. ROE showed excellent antibacterial activity against six pathogens of aquatic animals: Vibrio parahaemolyticus, V. vulnificus, V. alginolyticus, V. carchariae, Aeromonas hydrophila, and Edwardsiella tarda. The minimum inhibitory and bactericidal concentrations of measured ROE-derived anthraquinones were 10.57-84.53 µg/ml and 10.57-169.05 µg/ml, respectively.

Effect of curcumin on the expression of ahyI/R quorum sensing genes and some associated phenotypes in pathogenic Aeromonas hydrophila fish isolates.

Quorum sensing (QS) in Aeromonas hydrophila is mainly based on the modulation of ahyI/R genes that regulates bacterial virulence determinant phenotypes. The use of QS inhibitors would be of particular interest in inhibiting bacterial pathogenicity and infections. In this study, we aimed to determine the effect of curcumin, a natural component of Curcuma longa, on the expression of QS regulating genes, ahyI and ahyR, as well as some QS regulated virulence characteristics in pathogenic fish isolated A. hydrophila strains. The minimum inhibitory concentration (MIC) of curcumin against bacteria was determined using the broth micro-dilution method and the expression of quorum sensing genes ahyI and ahyR among the bacteria treated with curcumin was determined using quantitative polymerase chain reaction (qPCR). Also, the effect of curcumin on some QS associated traits, including biofilm formation, swarming and swimming motility, proteolytic potential, and bacterial hemolytic activity was investigated. According to the results, curcumin, at a concentration of 32 µg/mL, significantly reduced the expression of both ahyI and ahyR genes among bacterial strains up to 64.2 and 91.0%, respectively. Moreover, curcumin efficiently inhibited bacterial biofilm formation, swimming, and swarming motility. Also, bacterial proteolytic activity was slightly reduced, while hemolytic activity was not significantly affected. This study demonstrated the use of curcumin to attenuate ahyI/R QS genes and several QS associated phenotypes in A. hydrophila. These findings indicate the therapeutic potential of curcumin as an anti-QS agent, to be used against A. hydrophila pathogenesis in aquaculture.

The antioxidant power of horseradish, Armoracia rusticana, underlies antimicrobial and antiradical effects, exerted in vitro.

Armoracia rusticana (AR) was tested for antimicrobial and antioxidants power. The compound demonstrated to inhibit fish pathogens such as Vibrio anguillarum, V. harvey, V. alginolyticus, Aeromonas hydrophila, A. salmonicida, Photobacterium damselae subspecie piscicida, Tenacibaculum marinum and Pseudomonas anguilliseptica,. The total phenolic content and the reducing power resulted higher in the water extract of AR, respect to the hydroalcoolic. In vitro test demonstrated that AR significantly protect cells against death, induced by oxidative stress.

Study the immunomodulation of anthracenedione in striped dwarf catfish, Mystus vittatus against pathogenic bacteria, Aeromonas hydrophila.

Anthracenedione is a derivative of anthraquinone aromatic organic natural pigments found in senna, aloe latex, rhubarb, cascara, lichens, and fungi having broad range of bioactivity, including anti-cancer, anti-inflammatory, anti-microbial, anti-fungal, anti-oxidant, anti-viral activities suggesting potential for clinical purpose of many diseases. The effect of anthracenedione enriched diet on growth, hematology, innate and adaptive immune parameters as well as protection from Aeromonas hydrophila in Mystus vittatus was reported. The weight gain, feed intake, specific growth rate (SGR), and feed conversion ratio (FCR) were significantly increased in uninfected groups fed with 5 mg kg-1 diet. The red blood cells (RBC) and white blood cells (WBC) count and the percentage of lymphocytes were significantly augmented in both infected and uninfected groups feeding with any diet. The percentage of monocytes, eosinophils, neutrophils and the biochemical profile such as total protein, albumin, and globulin also were significantly increased in the infected and uninfected groups fed with 5 mg kg-1 enriched diet. The innate and adaptive immune parameters such as phagocytic activity, immunoglobulin M (IgM), respiratory burst activity, complement activity, and lysozyme activity were significantly increased in uninfected and infected groups fed with 5 or 10 mg kg-1 diets but not with 1 mg kg-1 diet. The serum superoxide dismutase (SOD) activity is significantly increased in the uninfected and infected fish fed with 5 mg kg-1 diet but the increase was not significantly observed in 1 or 10 mg kg-1 diets. The nitric oxide (NO) production is significantly elevated in both uninfected and infected groups fed with 5 mg kg-1 diet. On the other hand, the lymphocyte proliferation and myeloperoxidase (MPO) activity were significantly increased the infected and uninfected groups fed with 5 and 10 mg kg-1 diets. The cumulative mortality was found 5% with 1 and 5 mg kg-1 diet groups while it was observed 10% mortality with 10 mg kg-1 diet group. Based on the results, it is observed that feeding the uninfected and infected groups with 5 mg kg-1 anthracenedione diet resulted in better improvement of growth, hematological, biochemical, and innate as well as adaptive immune parameters in M. vittatus against A. hydrophila.

Effects of chitosan-supplemented diets on the growth performance, nonspecific immunity and health of loach fish (Misgurnus anguillicadatus).

This study investigated the effects of dietary chitosan on the growth performance and nonspecific immunity of loaches (Misgurnus anguillicadatus). Four practical diets at three levels of chitosan (1, 5 and 10 g/kg) were fed to loach (3.13 ±â€¯0.02 g) in triplicate groups (20 fish per replicate) for 10 weeks. Contrary to high-chitosan treatment, the administration of low or moderate levels of oral chitosan (1 or 5 g/kg) significantly increased the body weight gain, specific growth rate and condition factor. The oral administration of chitosan significantly increased the survival rate, phenoloxidase, superoxide dismutase, glutathione peroxidase, lysozyme, acid phosphatase and alkaline phosphatase activities, as well as the immunoglobulin M and complement 3 contents and the disease resistance against Aeromonas hydrophila. The optimum dose of dietary chitosan required for the maximum growth of loach was 5 g/kg. These results indicated that chitosan exerted immunostimulatory effects on loaches (M. anguillicadatus) and can thus be used as a dietary supplement.

Characterization of a polysaccharide HP-02 from Honeysuckle flowers and its immunoregulatory and anti-Aeromonas hydrophila effects in Cyprinus carpio L.

Aquaculture systems are known to be major reservoirs of Aeromonas hydrophila. The use of Chinese medicinal polysaccharides has become a successful strategy to prevent bacterial diseases in aquaculture. A water-soluble polysaccharide, HP-02, was purified from honeysuckle flowers, and the present study was aimed to examine its immunomodulatory and anti-A. hydrophila effects in Cyprinus carpio L. The results showed that the molecular weight of HP-02 was estimated to be 38 kDa. Monosaccharide composition was determined to be arabinose, rhamnose, mannose, glucose, and galactose in a molar ratio of 2.5: 1.8: 3.6: 3.7: 1.9. HP-02 not only distinctly improved the proliferation and phagocytosis of head kidney cells, but also exerted significant immunoregulation activity by increasing the secretion of the pro-inflammatory cytokines TNF-α, IL-1ß, IL-6, and IL-12 and the anti-inflammatory cytokines IL-10 and TGF-ß at the protein level in head kidney cells and serum. Moreover, during pathogen infection in vivo, HP-02 significantly reduced the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines. These results indicated that HP-02 had immunomodulatory effects on common carp both in vitro and in vivo, and HP-02 can be applied as a common carp feed supplement to enhance fish immunity and disease resistance against A. hydrophila.

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.

Antibacterial synergy between rutin and florfenicol enhances therapeutic spectrum against drug resistant Aeromonas hydrophila.

Emergence of antibiotic resistant bacteria has necessitated the drive to explore competent antimicrobial agents or to develop novel formulations to treat infections including Aeromonas hydrophila. The present study investigates the synergistic antibacterial effects of citrus flavonoid rutin and florfenicol (FF) against A. hydrophila in vitro and in vivo. Rutin is extracted and purified from Citrus sinensis peel through preparative HPLC and characterized through TLC, GC-MS and 1H and 13C NMR analyses. Though rutin did not display significant antibacterial activity, it modulated FF activity resulting in four-fold reduction in the MIC value for FF. The anti-biofilm potential of synergistic association of rutin and FF was validated by protein analysis, quantification of exopolysaccharide (EPS) and microscopy studies using sub-MIC doses. Besides antibacterial action, in vivo studies showed that Rutin/FF combination enhanced host immunity by improving blood cell count, anti-protease, and lysozyme activities as well as decreased the oxidative stress and the pathological changes of tilapia Oreochromis niloticus against A. hydrophila infection. No significant DNA damages or clastogenic effects were detected in tilapia challenged with A. hydrophila under Rutin/FF treatment. It is shown that an acute-phase Lipopolysaccharide binding protein (LBP) enhances the innate host defence against bacterial challenge. Semi quantitative RT-PCR and western blot results revealed the significant increase of LBP in the supernatant of tilapia monocytes/macrophages challenged with A. hydrophila upon treatment. The study findings substantiate that the combination of natural molecules with antibiotics may open up possibilities to treat MDR strains.

Potential KPC-2 carbapenemase reservoir of environmental Aeromonas hydrophila and Aeromonas caviae isolates from the effluent of an urban wastewater treatment plant in Japan.

Aeromonas hydrophila and Aeromonas caviae adapt to saline water environments and are the most predominant Aeromonas species isolated from estuaries. Here, we isolated antimicrobial-resistant (AMR) Aeromonas strains (A. hydrophila GSH8-2 and A. caviae GSH8M-1) carrying the carabapenemase blaKPC-2 gene from a wastewater treatment plant (WWTP) effluent in Tokyo Bay (Japan) and determined their complete genome sequences. GSH8-2 and GSH8M-1 were classified as newly assigned sequence types ST558 and ST13, suggesting no supportive evidence of clonal dissemination. The strains appear to have acquired blaKPC-2 -positive IncP-6-relative plasmids (pGSH8-2 and pGSH8M-1-2) that share a common backbone with plasmids in Aeromonas sp. ASNIH3 isolated from hospital wastewater in the United States, A. hydrophila WCHAH045096 isolated from sewage in China, other clinical isolates (Klebsiella, Enterobacter and Escherichia coli), and wastewater isolates (Citrobacter, Pseudomonas and other Aeromonas spp.). In addition to blaKPC-2 , pGSH8M-1-2 carries an IS26-mediated composite transposon including a macrolide resistance gene, mph(A). Although Aeromonas species are opportunistic pathogens, they could serve as potential environmental reservoir bacteria for carbapenemase and AMR genes. AMR monitoring from WWTP effluents will contribute to the detection of ongoing AMR dissemination in the environment and might provide an early warning of potential dissemination in clinical settings and communities.

Antibacterial activity of Indonesian red algae Gracilaria edulis against bacterial fish pathogens and characterization of active fractions.

In this study the crude ethyl acetate extracts and fractions from red algae Gracilaria edulis were studied against fish pathogens. G. edulis was extracted with ethyl acetate, fractionated in silica gel coloum chromatography and gradient hexane-ethyl acetate. The extract and fractions were subjected for bioassay guided isolation against fish pathogen Vibrio spp and Aeromonas hydrophila. Bioassays were accomplished by diffusion agar, bioauthography, and 96 well microplates format. Profiling chemical constituent was done by thin layer chromatography and GC-MS. Metabolites analysis using GC-MS yielded chemical constituents of active fractions as 9-hexadecenoic acid, hexadecanoic acid, 13-octadecenoic acid, 10-octadecenoic acid, eicosanoic acid, cholest-8-en-3-ol. The results provide in vitro scientific support to the possible application of Gracilaria edulis as antimicrobial agent that may contribute for treatment of bacterial infections in aquaculture.

Myo-inositol as an adjuvant to florfenicol against Aeromonas hydrophila infection in common carp Cyprinus carpio.

Florfenicol, a synthetic drug with chemical structure and spectrum of antibacterial activity similar to chloramphenicol, has been shown to be effective against a number of bacterial pathogens. However, there are increasing signs of florfenicol-resistant bacteria due to the misuse and overuse of florfenicol in aquaculture. In the present study, florfenicol had a higher bactericidal efficacy in the presence of myo-inositol, which may be due to the ability of myo-inositol to increase susceptibility of Aeromonas hydrophila to florfenicol. Furthermore, in two different infected models, co-administration of myo-inositol and florfenicol significantly reduced the bacterial load in the liver, kidney and spleen tissues of A. hydrophila-infected Cyprinus carpio, and greatly increased the survival rate of infected fish. Finally, it was also found that myo-inositol exhibited synergistic action with other antibiotic drugs including neomycin sulfate, ceftriaxone and enrofloxacin. The results obtained in this study suggest that myo-inositol as an efficient adjuvant to antibiotic drugs could be useful in increasing the antimicrobial activity of antibiotic drugs against A. hydrophila infection, and could also be useful to help decrease the occurrence of antibiotic overuse in aquaculture.

Ceftriaxone-Resistant Aeromonas hydrophila Infection Following Leech Therapy: A New Resistant Strain.

We report a case of ceftriaxone-resistant Aeromonas hydrophila infection in a 32-year-old man after medicinal leech therapy. The patient underwent a free gracilis muscle flap for dorsal foot soft tissue reconstruction. Because of persistent congestion after venous revision, medicinal leech therapy was initiated to aid in flap salvage. Intravenous ceftriaxone was administered prior to application of leeches for prophylaxis against A. hydrophila. Leech therapy was discontinued after 1 week, and ceftriaxone was discontinued 24 hours thereafter. An infection developed despite being on appropriate antibiotics. Culture sensitivities indicated that A. hydrophila was resistant to ceftriaxone.

Evaluation of In Vivo Probiotic Efficiency of Bacillus amyloliquefaciens in Labeo rohita Challenged by Pathogenic Strain of Aeromonas hydrophila MTCC 1739.

Diseases in aquatic organisms, including fish, are a major concern in aquaculture production. In this present investigation, we have evaluated the beneficial effects of dietary Bacillus amyloliquefaciens CCF7 (GenBank Acc. No. KP256501) supplementation in rohu (Labeo rohita) challenged by a pathogenic strain of Aeromonas hydrophila MTCC 1739. Four experimental diets were formulated: control diet (no probiotics) and three experimental diets (different concentrations of probiotic candidate B. amyloliquefaciens CCF7 at 105 (T1), 107 (T2), 109 (T3) CFU/g). Further, we have divided the feeding trial into pre-challenge (70 days) and post-challenge (28 days) periods and various immune parameters (serum protein, globulin, albumin, lysozyme, and IgM), and stress parameters (malondialdehyde, catalase, and superoxide dismutase) were examined during both the periods. Throughout the entire experiment, control group was fed with probiotic free basal diet, while the treatment groups received probiotic supplemented diets (PSD). After challenge test, serum aspartate transaminase (AST), serum alanine transaminase (ALT) activity, and liver malondialdehyde level have increased significantly in control groups; however, level of these parameters were considerably lower in fish fed with PSD. In contrast, liver catalase and superoxide dismutase activities and serum globulin concentration was significantly higher in the group fed with T3 diet followed by T2. Furthermore, an elevated level of serum IgM and higher activity of serum lysozyme was also recorded in PSD fed groups, especially for T3 group which confirmed the probiotic efficiency of the bacterium B. amyloliquefaciens CCF7. We strongly believe that B. amyloliquefaciens CCF7 will be a good probiotic candidate in aquaculture industries.

Activity of Brazilian propolis against Aeromonas hydrophila and its effect on Nile tilapia growth, hematological and non-specific immune response under bacterial infection

ABSTRACT The effect of the ethanolic extract of propolis (EEP) on Aeromonas hydrophila was analyzed by determination of minimum inhibitory concentration (MIC). Then, the effects of crude propolis powder (CPP) on growth, hemato-immune parameters of the Nile tilapia, as well as its effects on resistance to A. hydrophila challenge were investigated. The CPP (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0%) was added to the diet of 280 Nile tilapia (50.0 ± 5.7 g fish-1). Hemato-immune parameters were analyzed before and after the bacterial challenge. Red blood cell, hematocrit, hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and hydrogen peroxide (H2O2) and nitric oxide (NO) were evaluated. The MIC of the EEP was 13% (v/v) with a bactericidal effect after 24 hours. Growth performance was significantly lower for those fish fed diets containing 2.5 and 3% of CPP compared to the control diet. Differences in CPP levels affected fish hemoglobin, neutrophils number and NO following the bacterial challenge. For others parameters no significant differences were observed. Our results show that although propolis has bactericidal properties in vitro, the addition of crude propolis powder to Nile tilapia extruded diets does not necessarily lead to an improvement of fish health.

Activity of Brazilian propolis against Aeromonas hydrophila and its effect on Nile tilapia growth, hematological and non-specific immune response under bacterial infection.

The effect of the ethanolic extract of propolis (EEP) on Aeromonas hydrophila was analyzed by determination of minimum inhibitory concentration (MIC). Then, the effects of crude propolis powder (CPP) on growth, hemato-immune parameters of the Nile tilapia, as well as its effects on resistance to A. hydrophila challenge were investigated. The CPP (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0%) was added to the diet of 280 Nile tilapia (50.0 ± 5.7 g fish-1). Hemato-immune parameters were analyzed before and after the bacterial challenge. Red blood cell, hematocrit, hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and hydrogen peroxide (H2O2) and nitric oxide (NO) were evaluated. The MIC of the EEP was 13% (v/v) with a bactericidal effect after 24 hours. Growth performance was significantly lower for those fish fed diets containing 2.5 and 3% of CPP compared to the control diet. Differences in CPP levels affected fish hemoglobin, neutrophils number and NO following the bacterial challenge. For others parameters no significant differences were observed. Our results show that although propolis has bactericidal properties in vitro, the addition of crude propolis powder to Nile tilapia extruded diets does not necessarily lead to an improvement of fish health.

A novel probiotic Bacillus siamensis B44v isolated from Thai pickled vegetables (Phak-dong) for potential use as a feed supplement in aquaculture.

The use of probiotic bacteria to control bacterial infection in farmed fish is of clear practical interest. The aims of this study were to isolate and select a probiotic Bacillus sp. and to evaluate the effects of its supplementation on the growth and disease resistance of hybrid catfish. Bacillus siamensis strain B44v, selectively isolated from Thai pickled vegetables (Phak-dong), displayed a high potential as a probiotic in catfish culture. This bacterium produced a bacteriocin-like substance and exhibited a broad-spectrum antibacterial activity inhibiting both Gram-positive and Gram-negative bacteria, especially the fish pathogens Aeromonas hydrophila and Streptococcus agalactiae. The susceptibility to all 14 antibiotics tested implies its less possibility to be the antibiotic-resistant bacterium. Bacillus siamensis strain B44v possessed interesting adhesion properties, as shown by its high percentages of hydrophobicity (64.8%), auto-agglutination (73.8%), co-aggregation (67.2% with A. hydrophila FW52 and 63.5% with S. agalactiae F3S), and mucin binding (88.7%). The strain B44v survived simulated gastrointestinal conditions and produced protease and cellulase enzymes. Hybrid catfish (C. macrocephalus × C. gariepinus) were employed in the feed-trial experiments. Fish fed diet containing strain B44v (107 CFU/g feed) displayed not only no mortality but also growth improvement. At the end of the feed trial, fish were challenged by an intraperitoneal injection of Aeromonas hydrophila FW52. The Bacillus siamensis strain B44v fed fish survived (75.0%; p < 0.05) better than the controls (36.7%; p < 0.05) after a two week challenge. These collective results present for the first time the potential of Bacillus siamensis strain B44v for use as a bacterial probiotic in aquaculture.