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1.
Aeromonas hydrophila and Gyrodactylus cichlidarum are common pathogens that induce significant economic losses in farm‐reared Nile tilapia. Nowadays, the sudden appearance of fish mortalities was exaggerated due to mixed and multiple infections. During summer 2016, mass mortality among earthen pond‐farmed Nile tilapia was reported. Clinico‐pathological, bacteriological and parasitological examinations have been demonstrated. As well, the water quality parameters were assessed. The clinical and histopathological findings of the moribund and recently dead fish were characterized by generalized septicaemic signs. The water quality parameters were significantly elevated over the permissible levels, whereas there was an elevation in nitrite (0.04 mg/L), un‐ionized ammonia (0.8 mg/L), hydrogen sulphide levels (153.1 mg/L) and organic matter content (3.79 mg/L). A. hydrophila was identified based on phenotypic characterization, API 20E features and the homology of 16S rRNA gene sequence analysis. In addition, PCR data confirmed the presence of aerolysin (aerA) and haemolysin (hly) genes in the identified A. hydrophila isolates. Gene sequencing and phylogenetic analysis based on 16S rRNA sequence confirmed that A. hydrophila H/A (accession No. MN726928) of the present study displayed 98%–99% identity with the 16S rRNA gene of A. hydrophila. Furthermore, the monogenetic trematode, G. cichlidarum was identified in the wet mounts from the skin and gills of the examined fish with a high infestation rate. In this context, it was reported that the synergistic co‐infection of A. hydrophila and G. cichlidarum with deteriorated water quality parameters could induce exaggerated fish mortalities during hot weather.  相似文献   

2.
The outer membrane protein of Aeromonas hydrophila is a potential candidate for vaccine development. In this study, after cloning and expression of ompTS, 270 common carp, weighing 44 ± 5.7 g divided into five groups, were injected intraperitoneally twice with 3‐week intervals. Groups included the following: PBS, PBS plus Freund's adjuvant, recombinant protein, recombinant protein plus Freund's adjuvant and 20 fish as negative control. Two weeks after the second injection, 30 fish of each group were challenged with a dose of 2 × LD50 of Aeromonas hydrophila and RPS was measured. The antibody level was measured using ELISA test. The protection of recombinant protein in the immunized fish with and without adjuvant, respectively, was about 82.61% and 78.26% (the protection of recombinant protein electroeluted from an SDS–PAGE with and without adjuvant, respectively, was about 78.62% and 69.57%). The average of antibody level in recombinant protein with and without adjuvant was significantly higher than the PBS group (p < .05). The ability of recombinant ompTS to increase the antibody level and to protect the fish from challenge by A. hydrophila demonstrated that recombinant ompTS protein injection can be used to immunize common carp against A. hydrophila infection.  相似文献   

3.
Aeromonas hydrophila is a pathogen infecting farmed hybrid catfish, Clarias macrocephalus (Günther, 1864) × Clarias gariepinus (Burchell, 1822) which incurs substantial economic losses in Thailand. The study aimed at a genetic tracking of A. hydrophila infection and the in vitro assessment of the efficacy of antibiotics against its virulent strains. Five clinical strains from catfishes and Nile tilapia were employed. They were 3‐passage re‐isolated through healthy hybrid catfish and the cytolytic enterotoxin gene (AHCYTOEN) of individuals was traced. Each of the re‐isolates at a dose of ~6.67 × 105 CFU/g was intraperitoneally injected into ~15 g‐healthy hybrid catfish and their pathogenicity were observed for 7 days. It was found that AHCYTOEN was carried over whereas typical signs of motile aeromonas septicaemia were found in the specimens. The bacterial strains of Nile tilapia origin did not induce mortality but those of catfish origins (80%–100% rate of mortality). The strains were susceptible to the tetracycline antibiotics, and oxytetracycline produced MIC50 and MBC as low as 0.007–0.031 μg/ml and 1–8 μg/ml respectively. As oxytetracycline specifically inhibited pathogenic A. hydrophila in vitro, it is recommended that an appropriate dosage regimen of the drug should be established.  相似文献   

4.
The aim of this research is to evaluate the effects of biofloc system application at different fish density on the nursery production performance and the robustness of African catfish Clarias gariepinus (Burchell) against Aeromonas hydrophila and salinity stress. An economic analysis was also performed to determine the most optimum fish density in biofloc‐based African catfish nursery production. African catfish with an average body weight of 0.96 ± 0.04 g and average body length of 4.20 ± 0.46 cm, respectively, were distributed in 12 units of circular plastic‐lined tanks (1,020 L). The experiment consisted of four treatments in triplicates, that is, a control without carbon source addition at a density of 4 fish/L, and three biofloc treatments at different densities, that is, BFT4 (4 fish/L), BFT6 (6 fish/L) and BFT8 (8 fish/L). Tapioca flour was used as the organic carbon source in biofloc systems and was added at an estimated C/N ratio of 10. Housing the fish in biofloc systems resulted in higher fish growth, more efficient feed utilization, higher fish robustness against A. hydrophila infection and salinity stress, as well as higher profitability of nursery production. Increasing the fish density resulted in higher mortality. However, higher number of fish produced and lower feed conversion ratio were observed in the treatments with higher density (6 and 8 fish/L). In conclusion, the application of biofloc technology at a density of 8 fish/L could be recommended to increase the production and profitability of African catfish nursery culture.  相似文献   

5.
Aeromonas hydrophila is known to be causative agent of an infection named as Bacterial haemorrhagic septicaemia or red pest in freshwater fish. The aim of this study was to develop and validate the glycoprotein‐based fish vaccine against Aeromonas hydrophila. For this aim, after identification and characterization of A. hydrophila isolates from fish farms, one A. hydrophila isolate was selected as vaccine strain. Antigenic glycoproteins of this vaccine strain were determined by Western blotting and glycan detection kit. The connection types of these glycoproteins were examined by glycoprotein differentiation kit. Two glycoproteins, molecular weights of 19 and 38 kDa, with SNA connection type were selected for use in vaccination trials. After their purification by SNA‐specific lectin and size‐exclusion chromatography, protection studies with purified proteins were performed. For challenge trials, four experimental fish groups were designated: Group I (with montanide), Group II (with montanide and ginseng), Group III [with Al(OH)3] and Group IV [with Al(OH)3 and ginseng]. The survival ratings of fish were determined, and protection was calculated as 21.56%, 29.41%, 69.83% and 78.88% in groups I, II, III and IV, respectively. In conclusion, A. hydrophila glycoproteins with Al(OH)3 and ginseng could be used as a safe and effective vaccine for fish.  相似文献   

6.
Vibrio vulnificus, Edwardsiella anguillarum and Aeromonas hydrophila are three common bacterial pathogens in cultivated eels. To protect farming eels from infection by these pathogens, a trivalent outer membrane protein (OMP) containing partial sequences of OmpU from V. vulnificus, OmpA from E. anguillarum and OmpII from A. hydrophila was expressed and purified; then, the OMP was used as a vaccine to immunize Japanese eels (Anguilla japonica). Whole-blood cell proliferation, antibody titres and complement and lysozyme activities were detected at different days post-immunization (dpi), and the relative per cent survival (RPS) was determined after eels were infected with V. vulnificus, E. anguillarum or A. hydrophila at 28 dpi. The results showed that the OMP significantly stimulates the antibody titres. At 14 days after the challenge (i.e. at 28 dpi), the RPS of OMP against V. vulnificus, E. anguillarum and A. hydrophila was 20%, 70% and 11.1%, respectively. The construction, expression and immunogenicity of a trivalent Omp were reported for the first time, and this study will provide a valuable reference for the development of fish multiplex vaccines.  相似文献   

7.
An 84‐day feeding trial was carried out to evaluate the effects of aqueous Psidium guajava leaf extract (PGE) on growth, intestinal morphology, physiology, immune response and susceptibility of Oreochromis niloticus to Aeromonas hydrophila. Diets containing 0% (P0), 0.25% (P1), 0.50% (P2), 0.75% (P3) and 1.00% (P4) PGE were fed to triplicate groups of fish (mean weight; 1.32 ± 0.04 g) for 84 days. After the 84‐day feeding trial, test fish were injected with pathogenic A. hydrophila and then fed for 14 days. More feed were consumed in groups of fish fed PGE‐treated diets and resulted in significantly higher weight gain and feed intake. Incidentally, there was an increase in the calculated area of absorption of fish fed PGE diets, as accounted for by marginally higher villi length and width. Antioxidant and immune response were improved with PGE inclusion in diets as total protein, superoxide dismutase, glutathione peroxidase and glutathione S‐transferase significantly increased (p < 0.05) in fish fed PGE diets. Results of the challenge test with A. hydrophila revealed that the highest mortality (100%) was recorded in P0. This study revealed that inclusion of P. guajava extract in the diet of O. niloticus improved growth, nutrient utilization, immune system and survival of O. niloticus fingerlings.  相似文献   

8.
The two main diseases in the pangasius catfish industry are bacillary necrosis of Pangasianodon (BNP) and motile aeromonas septicaemia (MAS), where the aetiological agents have been identified as Edwardsiella ictaluri and Aeromonas hydrophila, respectively. In this study, apparently healthy Pangasianodon hypophthalmus were exposed to E. ictaluri, A. hydrophila or both bacterial species by intraperitoneal injection or immersion. There were 20 fish per treatment group, and the bacterial isolates used for the study were recovered from natural infections of BNP or MAS in farmed Vietnamese P. hypophthalmus. The results of the experimental infections mimicked the natural disease outbreaks reported from these pathogens in P. hypophthalmus. Furthermore, it was clearly demonstrated that E. ictaluri was only recovered from the fish exposed to the bacterium and not recovered from the animals receiving A. hydrophila.  相似文献   

9.
For the sustainable farming of tilapia, proper maintenance of their health and adequate treatment for infections at appropriate time are inevitable. The indiscriminate use of antibiotics in aquaculture, as a part of treatment and as growth promoters, accelerates antimicrobial resistance (AMR) among the fish pathogens. In the present study, we have isolated diverse aeromonads from Nile tilapia and studied their antibiogram and plasmid profiling. Aeromonas hydrophila, Aeromonas veronii, Aeromonas sobria, Aeromonas dhakensis, Aeromonas caviae, Aeromonas jandaei and Aeromonas aquatica were isolated from infected tilapia (n = 150), and their Shannon wiener diversity index was calculated as 1.926. A. veronii was found to be the most multiple antibiotic‐resistant pathogen with the MAR index of 0.46, and A. aquatica was noticed as the least resistant isolate. The minimum inhibitory concentration of resistant antibiotics was shown as >256 mcg/ml for most of the isolates. The virulent genes such as aerolysin and hemolysin were identified in all the isolates except A. aquatica. The detection of class 1 integrons, plasmid profiling and plasmid curing studies confirmed that AMR exhibited by most of the Aeromonas species is of plasmid mediated. This challenges the risk of wide spread of AMR among the pathogens and subsequent treatment of the infection.  相似文献   

10.
Outer membrane proteins (Omps) of Gram‐negative bacteria have been proven to be efficient subunit vaccines against bacteriosis. In this study, OmpF and OmpK of Aeromonas hydrophila were expressed, and their immune protective effects in European eel (Anguilla anguilla) were evaluated. The genomic DNA of A. hydrophila 322A was used as a template, and two kinds of prokaryotic expression plasmids, pET‐32a‐OmpF and pET‐32a‐OmpK, were constructed. Recombinant OmpF protein (r‐OmpF) and r‐OmpK were purified and were proven to have antigenicity by Western‐blot analysis. r‐OmpF and r‐OmpK were used as immunogens to immunize European eel by intraperitoneal injection. The mRNA expression of 6 immune‐related genes (IgM, IL‐10, IRF3, IRF7, LysG4 and HexB) in the liver tissues of eels at 1 hr, 3 hr, 6 hr, 12 hr, 24 hr, 72 hr and 10 days postimmunization was analysed by real‐time PCR. At 30 dpi, the serum antibody response was measured by ELISA. Fish were attacked at 15 dpi by live 322A to assess the protective immunity of r‐OmpF and r‐OmpK. All the six tested genes responded to r‐OmpF or r‐OmpK vaccination at varying degrees. The serum antibody titre of r‐OmpF‐ and r‐OmpK‐immunized groups was 1:1,600 and 1:3,200 respectively. In addition, r‐OmpF gave 35.5% of the relative immune protection rate to European eels, while r‐OmpK gave 70.0%. By analysing the protective immunity and the regulatory role in the immune‐related gene expression of the two recombinant proteins that were studied, it was found that r‐OmpK was a potential vaccine candidate against A. hydrophila.  相似文献   

11.
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12.
Experimental diets were formulated containing six levels of linolenic acid (0%, 0.5%, 1.0%, 1.5%, 2.0% and 2.5%) and each diet was fed to darkbarbel catfish Pelteobagrus vachelli in triplicate for 84 days. Weight gain, specific growth rate and feed efficiency increased with the increasing levels of dietary linolenic acid, but were not significantly different among diets containing 1.0%, 1.5%, 2.0% and 2.5% linolenic acid. Red blood cell count and haemoglobin were significantly higher in fish fed the 1.0% linolenic acid than other diets. Lysozyme activity and immunoglobulin M content in fish fed 1.0% of linolenic acid were significantly higher compared with the other diets. Antibody titre in fish fed 1.0–2.5% linolenic acids was significantly higher than in fish fed 0 or 0.5% linolenic acid 14 days after Aeromonas hydrophila challenge. This study indicates that linolenic acid is essential to darkbarbel catfish. Two‐straight broken‐line analysis with SGR showed that a dietary level of 1.29% linolenic acid is the minimum required for adequate growth of darkbarbel catfish. Considering other measured indicators of fish performance, a level of 1.0–1.5% linolenic acid in the diet is recommended for the darkbarbel catfish.  相似文献   

13.
Interrupting quorum sensing represents a novel anti‐infective strategy to combat bacterial pathogen, and biodegradation of quorum sensing signal AHLs has been proved to be an efficient way to control pathogenic Gram‐negative bacteria in aquaculture. In this study, the effect of Bacillus firmus sw40 as efficient AHL‐degrading strain on virulence of fish pathogen Aeromonas hydrophila and disease resistance of gibel carp Carassius auratus gibelio was investigated. The results demonstrated that in vitro the B. firmus sw40 extracellular production (ECP) was able to significantly decrease protease production, haemolytic activity and biofilm formation in A. hydrophila. Dietary administration of B. firmus sw40 (109 CFU/g) for 4 weeks significantly reduced the inflammatory cytokines TNF‐1a, TNF‐2a and IFN‐γ genes expression, antioxidant parameter MDA and GSH levels in serum and increased antioxidant enzyme SOD activity. Besides, B. firmus sw40 could significantly increase the survival of gibel carp with pathogenic A. hydrophila infection.  相似文献   

14.
Five N‐acyl homoserine lactone‐degrading bacteria (quorum quenching (QQ) strains) were selected to evaluate their impacts on growth, virulence factors and biofilm formation in Yersinia ruckeri in vitro. No difference was observed among the growth pattern of Y. ruckeri in monoculture and coculture with the QQ strains. To investigate the regulation of virulence factors by quorum sensing in Y. ruckeri, cultures were supplemented with 3oxo‐C8‐HSL. The results indicated that swimming motility and biofilm formation are positively regulated by QS (p < 0.05), whereas caseinase, phospholipase and haemolysin productions are not influenced by 3oxo‐C8‐HSL (p > 0.05). The QQs were able to decrease swimming motility and biofilm formation in Y. ruckeri. QQ bacteria were supplemented to trout feed at 108 CFU/g (for 40 days). Their probiotic effect was verified by Y. ruckeri challenge either by immersion or injection in trout. All strains could significantly increase fish survival with Bacillus thuringiensis and Citrobacter gillenii showing the highest and lowest relative percentage survival (RPS) values (respectively, 85% and 38%). Besides, there was no difference between the RPS values by either immersion or injection challenge expect for B. thuringiensis. The putative involvement of the QQ capacity in the protection against Yersinia is discussed.  相似文献   

15.
In the present study, the effects of Artemia supplemented with 2‐β‐mercapto‐ethanol (β‐ME) treated yeast cell, Saccharomyces cerevisiae, on growth and reproductive performance, lysozyme activity, and disease resistance to Aeromonas hydrophila of freshwater ornamental species, Poecilia latipinna, were investigated. Within 60 days, molly fish were fed with three treatments including commercial food (T1), un‐supplemented Artemia (T2), and Artemia supplemented with β‐ME‐treated yeast cell (at concentration of 4 × 107 CFU/L of water) (T3). After the feeding period, the fish were exposed to 100 μl of a suspension (1.1 × 107 cells/ml) of A. hydrophila (BCCM5/LMG3279) and the cumulative mortality rates were recorded for 12 days. No significant difference was found between survival rate and growth performance of P. latipinna except for weight gain that was higher in fish fed through Artemia supplemented with β‐ME‐treated yeast cell compared to control group. Fecundity rate was significantly improved in fish fed using T3 with the maximum amount of 49.5 ± 2.29 per female (p < 0.05). Besides, lysozyme activity was significantly increased in group 3 (p < 0.05). Moreover, lowest fish mortality was significantly observed in this treatment (p < 0.05). In addition, the number of colonies formed by yeast cell in T3 (634 × 103 CFU/g intestine) showed significant difference with other treatments (p < 0.05). In sum, Artemia enriched with β‐ME‐treated yeast improved reproductive indices, immune responses, and resistance against A. hydrophila of P. latipinna.  相似文献   

16.
A 120‐day feeding trial was conducted to assess the effects of dietary supplementation of zinc oxide (ZnO) and selenium (Se) nanoparticles (NPs) on growth, immunological and enzymatic profiles in rohu, Labeo rohita. Fishes were fed with basal diet incorporated with laboratory‐synthesized nanoparticles, viz., zinc oxide nanoparticles—10 mg/kg and Se nanoparticles—0.3 mg/kg throughout the experimental period. At an interval of every 30 days, treated and control fishes were randomly collected and subjected to estimation of growth, non‐specific immune parameters and serum enzyme assays. It was observed that there was significant (p < 0.05) increase in growth and non‐specific immune parameters like respiratory burst, lysozyme and myeloperoxidase activities as compared to the control group (only basal diet). Serum enzymatic profiles such as lactate dehydrogenase and alkaline phosphatase activities down‐regulated (p < 0.05) in treated group compared to control group, and superoxide dismutase and acetylcholine esterase activity up‐regulated in treated group. The relative percentage survival (RPS) found to be significantly higher in treated groups (60.00 ± 8.82%) as compared to control group (45.00 ± 6.17%) while challenged with virulent bacterial strain Aeromonas hydrophila. These results indicated that dietary supplementation of zinc oxide (ZnO) and selenium (Se) nanoparticles stimulates immunity and enhances resistance to bacterial infection in L. rohita.  相似文献   

17.
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18.
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).  相似文献   

19.
Streptococcus agalactiae is an important pathogen in fish, causing great losses of intensive tilapia farming. To develop a potential live attenuated vaccine, a re‐attenuated S. agalactiae (named TFJ‐ery) was developed from a natural low‐virulence S. agalactiae strain TFJ0901 through selection of resistance to erythromycin. The biological characteristics, virulence, stability and the immunization protective efficacy to tilapia of TFJ‐ery were determined. The results indicated that TFJ‐ery grew at a slower rate than TFJ0901. The capsule thickness of TFJ‐ery was significantly less (p < 0.05) than TFJ0901. When Nile tilapia were intraperitoneally (IP) injected with TFJ‐ery, the mortality of fish was decreased than that injected with TFJ0901. The RPS of fish immunized with TFJ‐ery at a dose of 5.0 × 107 CFU was 95.00%, 93.02% and 100.00% at 4, 8 and 16 weeks post‐vaccination, respectively. ELISA results showed that the vaccinated fish produced significantly higher (p < 0.05) antibody titres compared to those of control at 2 or 4 weeks post‐vaccination. Taken together, our results suggest that erythromycin could be used to attenuate S. agalactiae, and TFJ‐ery is a potent attenuated vaccine candidate to protect tilapia against S. agalactiae infections.  相似文献   

20.
The comparative effects of Freund's and Aloe vera gel as adjuvants on the expression of IL‐1β and TNF‐α genes were studied in vaccinated common carp (Cyprinus carpio) with Aeromonas hydrophila bacterin. Fishes were intraperitoneally immunized with A. hydrophila bacterin in combination with Aloe vera gel or Freund's and also without any adjuvant. At day 28 after immunization, all groups were challenged by lethal dose of A. hydrophila (107 cells/fish). Changes in the expression of IL‐1β and TNF‐α genes were evaluated in anterior kidney before challenge and 12, 24, 72 and 7 days postchallenge using quantitative real‐time PCR. Higher expression levels of both genes were observed in all vaccinated groups compared with non‐immunized group. Fishes which received Aloe vera gel showed higher expression of IL‐1β and TNF‐α in relation to animals which vaccinated with or without Freund's adjuvant. We concluded that Aloe vera gel in compared with Freund's adjuvant had a more stimulatory effect on the expression of immune‐related genes in vaccinated common carp and it can use as a novel adjuvant in aquaculture.  相似文献   

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