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1.
Flavobacterium columnare is the causative agent of columnaris disease in diverse fish species worldwide. Although columnaris is an important disease, the antimicrobial susceptibility pattern of F. columnare is not well studied. Thus, the purpose of this study was to test the in vitro antimicrobial susceptibility of 97 F. columnare isolates collected worldwide between 1987 and 2011 from 17 fish species. The broth microdilution technique was utilized for reliable testing of these fastidious organisms. None of the isolates displayed acquired resistance to florfenicol, gentamicin, ormetoprim‐sulfadimethoxine and trimethoprim‐sulfamethoxazole. Acquired resistance to chloramphenicol was detected in 1%, to nitrofuran in 5%, to oxytetracycline in 11% and to enrofloxacin, flumequine and oxolinic acid in 10%, 16% and 16% of the isolates, respectively, as reflected by a bimodal or trimodal distribution of their minimum inhibitory concentrations (MICs). One isolate showed acquired resistance towards several antimicrobial agents including erythromycin. Another isolate revealed acquired resistance towards – amongst others – ampicillin. The isolates displaying acquired resistance originated from ornamental fish species or Vietnamese catfish, except for two isolates coming from wild channel catfish in which acquired resistance was encountered towards oxytetracycline only. Fifty per cent of the resistant isolates from ornamental fish were shown to have acquired resistance against three classes of antimicrobial agents, assigning these isolates as multiple resistant. These data might indicate less prudent use of antimicrobials especially in ornamental fish species.  相似文献   

2.
Flavobacterium columnare, the causative agent of columnaris disease, causes substantial mortality worldwide in numerous freshwater finfish species. Due to its global significance and impact on the aquaculture industry continual efforts to better understand basic mechanisms that contribute to disease are urgently needed. The current work sought to evaluate the effect of L‐rhamnose on the growth characteristics of F. columnare. While we initially did not observe any key changes during the total growth of F. columnare isolates tested when treated with L‐rhamnose, it soon became apparent that the difference lies in the ability of this carbohydrate to facilitate the formation of biofilms. The addition of different concentrations of L‐rhamnose consistently promoted the development of biofilms among different F. columnare isolates; however, it does not appear to be sufficient as a sole carbon source for biofilm growth. Our data also suggest that iron acquisition machinery is required for biofilm development. Finally, the addition of different concentrations of L‐rhamnose to F. columnare prior to a laboratory challenge increased mortality rates in channel catfish (Ictalurus punctatus) as compared to controls. These results provide further evidence that biofilm formation is an integral virulence factor in the initiation of disease in fish.  相似文献   

3.
Flavobacterium columnare causes columnaris disease of farmed and wild freshwater fish. Skin mucus is an important factor in early stages of columnaris pathogenesis, albeit little studied. Our objectives were to (a) characterize the terminal glycosylation pattern (TGP) of catfish mucus, (b) determine the growth of F. columnare in formulated water (FW)‐containing channel catfish (Ictalurus punctatus) or hybrid catfish (Ictalurus punctatus X Ictalurus furcatus) mucus and (c) examine extracellular protease activity of two F. columnare isolates differing in virulence. The TGP of catfish mucus by lectin binding was as follows: alpha‐D‐mannose/alpha‐D‐glucose >N‐acetyl‐beta‐D‐glucosamine >N‐acetyl‐beta‐D‐glucosamine/N‐acetylneuraminic acid >N‐acetyl‐D‐galactosamine >alpha‐D‐galactose/N‐acetyl‐alpha‐D‐galactosamine >beta‐D‐galactose = alpha‐L‐fucose. Virulence studies demonstrated isolate AL‐02‐36 was highly virulent in channel catfish fry (0.1 g) with cumulative mortality of 90%‐100% versus 60% for isolate ALG‐00‐530 at equivalent doses (~3 × 106 CFU/ml); a similar result was observed in larger (0.7 g) catfish. In multiple experiments, F. columnare replicated (2‐3 logs) and survived (28 days) in formulated water‐containing catfish mucus. Highly virulent isolate AL‐02‐36 possessed at least 2.5‐ to fivefold higher protease activity following growth in mucus than the less virulent ALG‐00‐530. Flavobacterium columnare utilized catfish mucus as a nutrient source and mucus presence modulated extracellular protease production.  相似文献   

4.
Mucins are large glycoproteins that cover epithelial surfaces of the body and play important roles in prevention of inflammatory and various infectious diseases. In this study, five membrane‐bound and seven secreted mucin genes in the channel catfish were identified. All these identified mucin genes possess at least one PTS, von Willebrand D (VWD) or SEA domains. The expression of the 12 mucin genes in channel catfish was first studied with infection of Edwardsiella ictaluri and Flavobacterium columnare. Expression difference in MUC13a, MUC13, MUC2 and MUC5b was found in the intestine after E. ictaluri infection. Eight mucin gene expressions (except MUC3a, MUC2, MUC4 and MUC5f) were up‐regulated at 4 hr and down‐regulated after 24 hr in the gill with F. columnare infection. Expression level of MUC2 gene was up‐regulated in the intestine with E. ictaluri infection without no significant change in the gill under the F. columnare infection, which indicate that MUC2 is tissue‐specific gene expression and has different immune respond to two bacterial challenge. Taken together, the study showed mucin from the gill by F. columnare challenge induced an obvious response than mucin from the intestine with E. ictaluri infection.  相似文献   

5.
Columnaris disease, caused by the bacterial pathogen Flavobacterium columnare, continues to be a major problem worldwide in both wild and cultured freshwater finfish. Despite the far-reaching negative impacts of columnaris disease, safe and efficacious preventatives and curatives for this disease remain limited. In this study, we evaluated the potential of kaolin (Al2Si205(OH)4), a type of clay, for the prevention of columnaris disease. Channel catfish, Ictalurus punctatus (Rafinesque), fingerlings were experimentally challenged with Flavobacterium columnare in untreated water or with water containing kaolin (1 g L−1). Over the 7-day course of study, kaolin treatment led to significantly (P < 0.001) improved survival (96%) as compared to untreated fish (78% survival). Histological examination of the gills revealed that kaolin-treated fish had substantially less gill damage than untreated controls. Quantitative PCR analysis of gill tissue revealed that kaolin significantly reduced F. columnare adhesion (measured at 1 h post-challenge) and colonization (24 h post-challenge). Incubation of kaolin with F. columnare in vitro demonstrated that kaolin reduced the number of F. columnare cells in culture supernatants, presumably through the formation of physical complexes through adsorption. In summary, kaolin can improve survival, reduce gill pathologies and reduce bacterial attachment to key tissues associated with columnaris disease in channel catfish by binding to F. columnare.  相似文献   

6.
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7.
A feeding trial was performed to determine the effect of a commercial source of Yucca schidigera and Quillaja saponaria extracts (Nutrafito Plus®), in practical diets, on striped catfish Pangasianodon hypophthalmus growth, feed utilization, body composition, total ammonia‐nitrogen (TAN) excretion and haematological parameters. Four experimental diets were supplemented with Nutrafito Plus® at 0%, 0.01%, 0.02% and 0.03% (diets: control, N01, N02 and N03 respectively). Three replicate groups of striped catfish, with initial mean weight of 1.78 ± 0.05 g, were fed one of the four diets for 12 weeks. The specific growth rate (SGR) and final weight of fish fed diet N03 was significantly higher (P < 0.05) than fish fed the other dietary treatments. The growth performance of fish fed diets N01 and N02 were not significantly different compared to fish fed the control diet. Striped catfish fed diet N03 had improved feed conversion ratio (FCR), net protein utilization (NPU) and protein efficiency ratio (PER) than the control (P < 0.05). The inclusion of the high dietary level of Yucca schidigera and Quillaja saponaria, diet N03, reduced TAN compared to all groups. Dietary inclusion of Yucca schidigera and Quillaja saponaria at all levels investigated did not affect the whole body proximate composition of the striped catfish (P > 0.05). The packed cell volume (PCV) and haemoglobin level in fish fed diet N03 was significantly higher than in the fish fed the control diet. The present study demonstrates that dietary inclusion of Yucca schidigera and Quillaja saponaria induced positive effects on growth performance and haematological parameters and decreased TAN excretion in striped catfish.  相似文献   

8.
The efficacy of copper sulphate (CuSO4) against the early stages of an experimental acute infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus) was evaluated. Fish were challenged, by waterborne exposure to F. columnare in an ultra‐low flow‐through water delivery system, and treated with CuSO4 at 4.2 and 2.1 mg L?1 at 5.5 h post challenge for 4 h. Bacterial challenged non‐treated fish acted as a positive control and non‐challenged non‐treated fish acted as a negative control. Fish challenged with F. columnare and treated with CuSO4 at 4.2 and 2.1 mg L?1 post challenge had mortalities of 6.4% and 18.3%, respectively, compared with the positive control, which had 90.2% mortality. The mortality of challenged fish treated with CuSO4 at 4.2 and 2.1 mg L?1 was significantly different from the positive control. There was no significant difference between the mortalities of the 4.2 and 2.1 mg L?1 treatments. The results suggest that CuSO4 has clear therapeutic value against columnaris infections.  相似文献   

9.
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10.
The gliding aquatic bacterium Flavobacterium columnare causes columnaris disease, a common problem for wild and farmed freshwater fish worldwide. Recently, a broth microdilution method was standardized to test the susceptibility of F. columnare against antimicrobials commonly used in aquaculture. We used this new method to measure the minimal inhibitory concentrations (MICs) of ten antimicrobials against 120 F. columnare isolates. The resulting MIC frequency distributions for each antimicrobial (1 MIC/isolate) were used to estimate epidemiological cut‐off values (ECVs) which separate isolates with typical wild‐type (WT) susceptibility from isolates with decreased non‐wild‐type (NWT) susceptibility. We identified 22 NWT isolates with elevated MICs relative to the ECV that covered 99.9% of the MIC distribution against one or more of the antimicrobials: ampicillin, enrofloxacin, erythromycin, florfenicol, flumequine, oxolinic acid or oxytetracycline. Ten of the NWT isolates had decreased susceptibility to a single antimicrobial class, six isolates to two antimicrobial classes and six isolates to three or more antimicrobial classes. The MIC frequency distributions and provisional cut‐off values provide data needed to set epidemiological cut‐off values to monitor for the development of antimicrobial resistance among F. columnare.  相似文献   

11.
Numerous isolates of Flavobacterium columnare were previously recovered from red tilapia, Oreochromis sp., exhibiting columnaris‐like disease in Thai farms, and the phenotypic and genetic characteristics were described. The objective of this study was to determine the virulence of two morphotypes (rhizoid and non‐rhizoid colonies) of F. columnare and to determine their ability to adhere to and persist in red tilapia fry. The results showed that the typical rhizoid isolate (CUVET1214) was a highly virulent isolate and caused 100% mortality within 24 h following bath challenge of red tilapia with three different doses. The non‐rhizoid isolate (CUVET1201) was avirulent to red tilapia fry. Both morphotypes adhered to and persisted in tilapia similarly at 0.5 and 6 h post‐challenge as determined by whole fish bacterial loads. At 24 and 48 h post‐challenge, fry challenged with the rhizoid morphotype exhibited significantly higher bacterial loads than the non‐rhizoid morphotype. The results suggested that an inability of the non‐rhizoid morphotype to persist in tilapia fry may explain lack of virulence.  相似文献   

12.
A feeding trial with striped catfish, Pangasianodon hypophthalmus was performed to determine the effect of Yucca schidigera in practical diet on the growth, feed utilization, body composition, total ammonia–nitrogen (TAN) excretion and haematological parameters. A diet with fish meal as the main protein source without yucca extract was used as the control diet (Diet 1). Four diets were formulated with 0.075 (Diet 2), 0.1 (Diet 3) and 0.15 (Diet 4)% of yucca extract respectively. Fifteen fish per tank (initial mean weight 1.78 ± 0.05 g) were randomly allocated to 15 fibreglass tanks (80‐L) connected to a freshwater closed recirculation system (temperature 29.7 ± 1.0°C). The experimental diets were tested in triplicates for 12 weeks. The specific growth rate of fish fed Diet 4 was significantly higher when compared with fish fed Diet 1. The growth of fish fed diets Diet 2 and Diet 3 were not significantly different compared with fish fed the Diet 1. Striped catfish fed Diet 4 had significantly lower feed conversion ratio compared with fish fed Diet 1 and Diet 2 (P < 0.05).The incorporation of high level Yucca schidigera extract in the diets reduced TAN compared with Diet 1. Dietary inclusion of Yucca extract levels did not significantly affect the biometric parameters or whole body proximate composition of the striped catfish (P > 0.05). The PCV (%) in fish significantly increased with high levels of Yucca inclusion (Diet 4) compared with control diet. Fish fed Diet 4 showed significantly higher haemoglobin levels than Diet 1 (P < 0.05). The results indicate that dietary inclusion of Yucca schidigera extract is promising as a feed additive that could improve growth performance and some haematological parameters and the best Yucca schidigera level was 0.15%.  相似文献   

13.
Lipopolysaccharide (LPS), a component of Gram negative bacteria, was reported as important immunostimulant for fish. In this study, striped catfish were fed diets containing different Escherichia coli LPS concentrations (0%, 0.01% and 0.05%) for 2 weeks and then fed control feed (0% LPS) for 4 weeks. Plasma cortisol and glucose were rather low and did not differ significantly among treatments (P > 0.05). The respiratory burst activity, lysozyme, complement, total of antibody as well as mortality in fish challenged with Edwardsiella ictaluri were recorded every 2 weeks (W2, W4 and W6). The lysozyme activity significantly increased in fish treated with LPS (P < 0.05) in W2, W4 and W6. The highest values of respiratory burst activity were observed at week 4 in fish fed 0.01% LPS. There were significant differences in total of antibody between fish fed LPS (0.01%) and control in W2, W4. The challenge test with Edwardsiella ictaluri showed that fish fed 0.01% LPS had lower cumulative mortality (40%, 33% and 42%) compared with the fish fed 0.05% LPS (50%, 40% and 47%) and control fish (40%, 57% and 53%) in the three difference sampling times respectively. These results suggest that feed supplemented with 0.01% LPS could enhance immunity of striped catfish after 2 weeks of oral administration and fish could be protected against bacterial infection during the following 4 weeks.  相似文献   

14.
The influence of dietary supplementation of Lactobacillus acidophilus, on growth performance, digestive enzyme activities, gut histomorphology and gut microflora were evaluated in juveniles striped catfish (Pangasianodon hypophthalmus). Five experimental diets were formulated by supplementing 0, 103, 105, 107 and 109 CFU/g L. acidophilus in fishmeal and casein‐based semi‐purified diet. Triplicate groups of striped catfish (21.69 ± 0.18 g) were stocked in 15 fiberglass tanks with stocking density of 25 individuals per tank and fed twice daily at 2.5% of the fish body weight for 12 weeks. Weight gain, specific growth rate, feed conversion ratio and the protein efficiency ratio were significantly higher in fish fed with 105 and 107CFU/g L. acidophilussupplemented diets compared with the other treatment groups. Compared with the control and fish fed low (103 CFU/g) L. acidophilus supplementation, those fed with 105 and 107 CFU/g had significantly higher (p < 0.05) apparent protein digestibility. Inclusion of L. acidophilus at 107 CFU/g diet significantly increased amylase, protease and lipase activities. Microscopic analysis showed that the villi length in both the anterior and posterior gut and microvilli length in the posterior gut increased significantly in fish fed L. acidophilus supplementation at 105 and 107 CFU/g of diet. The fish fed L. acidophilus supplemented diets significantly increased the total lactic acid bacteria counts in the gut of striped catfish compared with the control‐fed group. Based on gut histomorphology and growth performance, inclusion of L. acidophilus at 105 CFU/g appears to have the most positive effect on striped catfish farming.  相似文献   

15.
A feeding trial was conducted to evaluate the potential of dietary supplementation of autolysed brewer's yeast (AY) on African catfish. The catfish (22.5 ± 1.15 g/fish, 20 fish 33 L/tank) were fed with either of diets (390 g/kg crude protein, 140 g/kg lipid) supplemented with 0, 3, 6 or 10 g/kg AY (n = 3). After 49 days of feeding, the final body weight and metabolic growth rate of the catfish fed 3 g/kg AY (3‐AY) diet were higher than those fed the control diet (p < .05). The lowest level (p < .05) of alanine transaminase was detected in the blood of the catfish fed 3‐AY diet. The mid‐intestinal histology of the catfish revealed no significant difference (p > .05) in intestinal perimeter ratio. However, an elevated (p < .05) abundance of goblet cells and intraepithelial leucocytes were found in the intestine of catfish fed 3, 6 and 10 g/kg AY diets, with the highest level of abundance recorded in the mid‐intestine of the catfish fed 3‐AY diet. The results suggest that dietary 3 g/kg autolysed brewer's yeast supplementation improves growth performance of African catfish without deleterious effect on liver functionality and gut morphology.  相似文献   

16.
The most common bacterial diseases in pond‐raised channel catfish Ictalurus punctatus (Rafinesque) are enteric septicemia of catfish and columnaris, caused by Edwardsiella ictaluri and Flavobacterium columnare respectively. Medicated feed containing antibiotics is one management approach that catfish producers use in the treatment of bacterial diseases. However, the future use of all types of medicated feed in catfish aquaculture is uncertain. To discover effective alternatives to antibiotics, a rapid 96‐well microplate bioassay utilizing E. ictaluri and F. columnare to evaluate natural compounds and extracts was developed. In this bioassay, bacterial growth is determined by absorbance measurements of microplate wells after 24 h incubation and then confirmed by detecting cell viability after the addition of 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide with additional incubation for 24 h. The minimum inhibitory concentration, minimum bactericidal concentration and 50% inhibition concentration (IC50) are determined by graphing the absorbance data. The 24 h IC50 results of test compounds are compared with the 24 h IC50 results of the drug controls oxytetracycline and florfenicol. Among the antibiotics evaluated, doxycycline and tetracycline appear more effective against E. ictaluri and F. columnare than either drug control. This bioassay is rapid, reproducible and economical for evaluating a large number of compounds and extracts.  相似文献   

17.
Effects of various concentrations of Scutellaria baicalensis (SB) extract in diets on growth, body composition, serum chemistry and disease challenge test of far eastern catfish (Silurus asotus) were determined and compared with a commercially available immune enhancer. Eight experimental diets were prepared in triplicate: control (Con) diet without supplementation of SB and SB‐0.25, SB‐0.5, SB‐1, SB‐2, SB‐3 and SB‐5 diets containing SB at concentrations of 0.25, 0.5, 1, 2, 3 and 5%, respectively. In addition, 0.1% of a commercial immune enhancer product (CP) was also tested. No significant difference in weight gain of fish was found. Feed consumption, feed efficiency ratio and protein retention of fish were not affected by the experimental diets. At the end of the 8‐week feeding trial, 10 externally normal fish from each tank were infected by Vibrio anguillarum or Strepotococcus iniae. Cumulative mortality of fish fed the Con diet was higher than that of fish fed the all other diets in 10 and 25 days after V. anguillarum or S. iniae infection. Results of this study indicated that dietary inclusion of SB extract was effective in improving survival of eastern catfish after V. anguillarum and S. iniae infection, but the various concentrations of SB did not affect fish performance.  相似文献   

18.
Columnaris disease is a major cause of mortality in tilapia hatcheries and commonly occurs during the summer season in Thailand. One way of reducing the problem is by selective breeding for increased disease resistance. The objective of this study was to estimate quantitative genetic parameters for resistance against columnaris in the Chitralada 4 strain of Nile tilapia. Data from 43 full‐sib families (2,580 records) of fry (age = 32 ± 4 days post‐hatch) were used in the analyses. Initially, fry were subjected to bath challenge with Flavobacterium columnare (LD50 concentration = 1.2 × 106 CFU/ml) for 14 days. Disease resistance was defined as the number of days from challenge until death (DD) or as a binary trait (dead/alive) on day 14. Linear animal and sire‐dam models were used for DD, while threshold animal, threshold sire‐dam, binary linear animal and binary linear sire‐dam models were used for binary outcomes. Covariate effect of age, fixed effect of challenge day and random effects of the individual animals or sires and dams were included in the models. Mean survival was 32.4 ± 11.6%, and survival rates of the best and poorest families were 70% and 8%, respectively. The highest estimate of heritability (0.30 ± 0.025) was obtained under the threshold sire‐dam model. Heritability estimates for DD (0.16 ± 0.034 and 0.17 ± 0.046) were comparable to those obtained from the threshold animal (0.15 ± 0.031) and the binary linear (0.14 ± 0.045 and 0.15 ± 0.044) models. The linear animal and sire‐dam models for DD and the threshold sire‐dam models performed equally with similar values of rEBV (0.629, 0.628 and 0.627) and accuracy of selection (0.793, 0.793 and 0.791). This study reveals the potential of selective breeding to increase disease resistance to F. columnare in the studied population of Nile tilapia.  相似文献   

19.
4‐Nonylphenol (NP) is an emerging concern contaminant which is widely spread in the aquatic ecosystem. Nigella sativa seed (NSS) has multifaceted therapeutic values. This study aimed to give insight into the potential protective effect of NSS on NP‐induced haematotoxicity in Clarias gariepinus through evaluation of haematological parameters, oxidant/antioxidant balance of blood lysate and histopathological investigation of blood smear. One hundred and fifty fish were divided into five groups (30/group). First group served as control which did not received NP exposure and fed basal diet without NSS supplementation. The other four groups were exposed to NP at a dose of 0.1 mg L?1 and fed diets supplemented with NSS at levels of 0, 10, 25 and 50 g/kg diet, respectively. Macrocytic hypochromic anaemia, thrombocytopenia, leucopenia, neutrophilia, lymphopenia, monocytosis and eosinophilia were observed following NP exposure together with increase in morphological erythrocyte alterations and micronuclei formation. Elevation in total peroxide and malondialdehyde and depletion in total antioxidant capacity of blood lysate were reported. We concluded that supplementation of NSS markedly ameliorated the previously listed manifestations, and the most effective doses were 25 and 50 g/kg feed.  相似文献   

20.
Flavobacterium columnare is divided into three genetic groups or genomovars, genomovar II being highly virulent for channel catfish. A modified live vaccine is currently available to prevent columnaris disease under the licensed name Aquavac‐Col®. The strain of F. columnare used to generate the avirulent rifampicin‐resistant mutant used in Aquavac‐Col® belonged to genomovar I, the less virulent group towards channel catfish. In this study, we describe the generation and characterization of rifampicin‐resistant mutants from genomovar II strains. A total of 13 new mutants were obtained, and eight of them (two from each parent strain) were genetically and phenotypically characterized. Highly conserved regions within the ribosomal operons were identical between parent and mutant strains. Genetic differences between mutants and their parent strains were revealed by amplified fragment length polymorphism (AFLP). Genetic changes were distinctive among different mutants. Analysis of the lipopolysaccharide (LPS) showed that while some mutants lacked a few molecular bands of the LPS, some exhibited the same LPS profiles as their parent strains. Comparison between immunogenic proteins from mutants and parents was carried out by immunoblot analysis and further confirmed the uniqueness of individual mutants. A complete set of rifampicin‐resistant mutants with different genetic and immunogenic properties from the highly virulent genomovar II has been created. These mutants may have the potential of becoming vaccine candidates against columnaris disease.  相似文献   

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