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1.
This study was conducted to evaluate the prevalence, potential pathogenicity and antimicrobial resistance of Vibrio isolates from 65 soil/water/fish samples collected from inland saline aquaculture areas. Depending on the sample type, presumptive Vibrio counts ranged from 2.50 to 6.16 log10 CFU/ml (or/g). Among the 119 confirmed Vibrio isolates, Vibrio cholerae was found to most dominant (91.6%) and it was detected in all the samples from inland saline areas. Seven other Vibrio spp. including Vibrio parahaemolyticus and Vibrio vulnificus were also detected. Except one O139 serotype, rest of the V. cholerae isolates were found belonging to non‐O1/non‐O139 serogroups. None of the V. cholerae isolate was found positive for ctx gene. Antimicrobial susceptibility testing against 7 commonly used antibiotics revealed highest resistance (50.4%) against ampicillin. Very high intermediate resistance (87.4%) was also observed against erythromycin. Contrary to previous studies, high susceptibility (>70%) to chloramphenicol, nalidixic acid, tetracycline and trimethoprim was observed in Vibrio isolates obtained in present study. Almost 20% of Vibrio isolates were resistant to two or more antibiotic classes with multiple antibiotic resistance (MAR) index value of ≥0.28. Presence of V. cholerae isolates with very high MAR index value of 0.85 also suggested that these multidrug‐resistant environment isolates could serve as reservoir of antibiotic‐resistant genes in aquatic systems. The presence of multiple drug resistance vibrios in emerging inland saline aquaculture systems emphasizes the need for their routine monitoring for developing the risk assessment and mitigation strategies.  相似文献   

2.
ABSTRACT

Vibrio parahaemolyticus is a human pathogen frequently found in seafood. Once the seafood is contaminated by V. parahaemolyticus, it can become a vehicle for foodborne illness. The conventional culture methods for detection of V. parahaemolyticus are time-consuming and cannot differentiate pathogenic strains from nonpathogenic ones. In this study, a multiplex polymerase chain reaction (PCR) technique was investigated for detecting tdh, chiA, and toxR of V. parahaemolyticus. The sensitivity of the multiplex PCR was determined by testing 28 strains of V. parahaemolyticus, 15 non-V. parahaemolyticus strains, and fresh seafood spiked with cells of V. parahaemolyticus. All the strains were analyzed for production of thermostable direct hemolysin (TDH) and chitinase. This study showed that both the chiA and toxR are excellent markers for detecting V. parahaemolyticus strains, and a multiplex PCR targeting chiA and tdh genes can be applied to simultaneously detect environmental and pathogenic V. parahaemolyticus.  相似文献   

3.
The polysaccharide – fucoidan was extracted from brown seaweed Sargassum wightii and its antibacterial activity was screened by agar well diffusion method. The maximum zone of inhibition observed was 15.66 mm in 20 mg mL?1 concentration against Vibrio parahaemolyticus. The Minimum Inhibitory Concentration (MIC) of the fucoidan was 12 mg mL?1 against V. parahaemolyticus. The fucoidan was then enriched with Artemia nauplii at four different concentrations such as 100, 200, 300 and 400 mg L?1 for 12 h. The enriched Artemia nauplii were fed to Penaeus monodon post‐larvae for 20 days and the growth performance was assessed. The weight gain and SGR of the control group were 0.2432 g and 15.78%, respectively. But, in experimental groups fed with fucoidan enriched Artemia nauplii, the weight gain and SGR were increased and were respectively ranged from 0.2602 to 0.3161 g and from 16.11 to 17.05%. The P. monodon post‐larvae were challenged with V. parahaemolyticus for a period of 30 days showed a reduction in mortality percentage of experimental groups over the control group and it was ranged between 36.97 and 89.86%. During the challenge test, the V. parahaemolyticus load was also enumerated from the infected shrimp at every 10 day intervals. In the control group, the Vibrio load showed a linear increase in hepatopancreas and muscle tissues from 10th to 30th days of challenge test, whereas in the experimental groups, the Vibrio load established a declining trend with the advancement of challenged test.  相似文献   

4.
Japanese flounder, Paralichthys olivaceus (Temminck et Schlegel), were immunized by the intraperitoneal (i. p.) injection route with formalin‐killed whole cells of Vibrio anguillarum that originated from a diseased fish. Fifty days later, a booster vaccination was given by the same route. Control fish were similarly treated with sterile phosphate‐buffered saline. The efficacy of vaccination was evaluated based on protection against two bacterial challenges and immune responses (both specific and non‐specific). The challenges were performed by i. p. injection with V. anguillarum or V. parahaemolyticus. The results indicated that the vaccinated fish showed higher non‐specific immune activity than the unvaccinated fish. The effects of vaccinations on the phagocytic activity of phagocyte, bactericidal and lysozyme activities were notable, especially on bactericidal activity. Determined by ELISA, antiserum of vaccinated fish displayed high antibody titres. The vaccination conferred protection against V. anguillarum challenge (81.25–93.75% relative percentage survival (RPS)). The RPS was 46.15–53.85% against V. parahaemolyticus challenge, indicating some degree of cross‐protective immunity.  相似文献   

5.
Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.  相似文献   

6.
Vibrionaceae infections are a major obstacle for marine larviculture; however, little is known about virulence differences of Vibrio strains. The virulence of Vibrio strains, mostly isolated from vibriosis outbreaks in farmed fish, was tested in larval challenge trials with cod (Gadus morhua), turbot (Scophthalmus maximus) and halibut (Hippoglossus hippoglossus) using a multiwell dish assays with single‐egg/larvae cultures. The strains differed significantly in virulence as some caused a high mortality of larva reaching 100% mortality after a few days, while others had no or only marginal effects on survival. Some Vibrio strains were pathogenic in all of the larva species, while some caused disease only in one of the species. Twenty‐nine of the Vibrio anguillarum strains increased the mortality of larvae from at least one fish species; however, pathogenicity of the strains differed markedly. Other Vibrio species had no or less pronounced effects on larval mortalities. Iron uptake has been related to V. anguillarum virulence; however, the presence or absence of the plasmid pJM1 encoding anguibactin did not correlate with virulence. The genomes of V. anguillarum were compared (D. Castillo, P.W. D'Alvise, M. Middelboe & L. Gram, unpublished data) and most of the high‐virulent strains had acquired virulence genes from other pathogenic Vibrio.  相似文献   

7.
Vibrio harveyi infection occurred with a moderate mortality in tiger puffer, Takifugu rubripes (Temminck et Schlegel), in autumn 2007, at a mariculture farm in western Japan. The diseased fish showed nodular lesions in the branchial chamber and the inner surface of the operculum. Histopathologically, the lesions comprised granulation tissue containing many suppurative foci allowing propagation of the bacteria and granuloma encapsulating abscesses with a decrease in bacteria. The bacteria were disseminated in visceral organs including the spleen, kidney, liver, and myocardium, resulting in the formation of granulomatous lesions. Two groups of tiger puffer juveniles were artificially infected by an intramuscular injection with an isolate (1.0_108 CFU/fish). During the experimental period, 20% mortality occurred within 4–6 days post‐infection (d.p.i). The fish sampled on 4 d.p.i showed abscesses in the lateral musculature at the injection site. The fish sampled 5 d.p.i. displayed the production of granulation tissue containing many suppurative foci, which replaced the necrotic dermis and lateral musculature. Surviving fish (15 d.p.i.) had granulomatous lesions in the lateral musculature at the injection site. Pyogranulomatosis is pathognomonic in V. harveyi infection of tiger puffer.  相似文献   

8.
Vibrio parahaemolyticus causes acute human gastroenteritis and is often linked to consumption of raw oysters. Previous investigations indicated that refrigerated seawater depuration at 12.5°C could significantly reduce V. parahaemolyticus contamination in Pacific oysters; however, further optimization is necessary to achieve the regulatory target of >3.52 log most probable number (MPN)/g reduction. The current study investigated influences of algal feeding on efficacy of depuration to reduce V. parahaemolyticus in raw oysters. A V. parahaemolyticus cocktail (10290, 10292, 10293, BE 98–2029, 027-1c1) was mixed in artificial seawater (70 L) to inoculate oysters (n = 35) at 4–5 log MPN/g. Inoculated oysters were subjected to depuration with feed (algae = 0.036 ml/gram of oyster) and without feed at 12.5°C. Oysters (n = 5) were analyzed for V. parahaemolyticus using a three-tube MPN method after 0, 1, 3, 5, and 6 days of depuration. Depuration (6 days) achieved average V. parahaemolyticus reductions of 2.75 log MPN/g and 3.03 log MPN/g in the fed and unfed systems, respectively; however, feeding status did not significantly impact the efficacy of depuration to reduce V. parahaemolyticus in Pacific oysters. Further optimization of depuration is necessary to achieve the regulatory target for V. parahaemolyticus decontamination in raw oysters.  相似文献   

9.
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10.
Species of the genus Vibrio have been recognized as one of the most significant pathogens in aquaculture farming, causing mass mortality of farmed stocks. Photodynamic Antimicrobial Chemotherapy (PACT) with singlet oxygen (1O2) has been identified as a powerful and sustainable water treatment method for pathogen eradication. In this study, the efficiencies of photolytic and photodynamic disinfection protocols were studied with two Vibrio species, Vibrio parahaemolyticus and Vibrio owensii. The selected microorganisms were successfully cultivated in marine broth and irradiations were performed with ~108 bacteria mL?1. Treated samples were monitored for bacterial regrowth for up to 7 days. Photolysis experiments were initially conducted with UV‐A, UV‐B for up to 2 h and visible (VIS) light for up to 24 h. Of these, only irradiation with UV‐B light for at least 45 min was efficient in controlling Vibrio. Irradiations with VIS light were subsequently repeated under PACT conditions in dose?response experiments with two water‐soluble porphyrins, [T4(MePy)P] and [TPPS4]. Disinfections of samples were successful for both porphyrin types at minimum concentrations of 10 μM and 24 h of irradiation.  相似文献   

11.
In this paper, we present a selective and differential medium, termed Vibrio alginolyticus (VAL) agar, developed for the isolation and identification of V. alginolyticus. The presence of bile salts, high salinity and high incubation temperature allows the selective growth of moderately halophilic Vibrio species. Differentiation of bacteria is achieved by identifying species capable of sucrose fermentation, made visible by the pH indicator bromocresol purple. In this study, all of the 26 strains of V. alginolyticus and only three of the 99 strains representing 30 species (including 19 Vibrio species) other than V. alginolyticus were able to grow in the VAL medium. The remaining three strains could be further differentiated from V. alginolyticus according to colour or the diameter of colonies produced on VAL agar plates. Colonies isolated from shellfish rearing water and infected shrimp through the use of VAL agar plates were all positively identified as V. alginolyticus by conventional tests and 16S rDNA sequencing. The testing of specificity and differentiation capability of VAL shows the potential of the agar as a medium for the primary isolation of V. alginolyticus from pathological and environmental samples.  相似文献   

12.
Diseases of shrimp have contributed to billions of dollars of economic loss in the aquaculture industry. Newly emerging strains of the bacterium Vibrio parahaemolyticus produce a condition in shrimp called early mortality syndrome or acute hepatopancreatic necrosis disease. Three different V. parahaemolyticus strains were evaluated for their respective pathogenicity on shrimp, Litopenaeus vannamei, when the bacterial strains were grown under various laboratory conditions prior to inoculating shrimp. For each trial, feed was inoculated with a known concentration of bacteria and then fed to the shrimp. The early mortality syndrome strain of V. parahaemolyticus was the most lethal resulting up to 100% mortality within 24 h after being introduced to shrimp via a single feeding. The other two strains of Vibrio, one isolated from the environment and the other from a human clinical case, resulted in 0% and 30% mortality within 96 h respectively. The concentration of the early mortality syndrome strain of V. parahaemolyticus that the shrimp were exposed to directly correlated with mortality rate, which allowed for lethal or sublethal short‐term disease challenge assays to be established. Infiltration of haemocytes was also evident in the midgut caeca of shrimp infected with the early mortality syndrome strain of V. parahaemolyticus, which has not been previously reported.  相似文献   

13.
Abstract

During the present investigation, laboratory experiments were conducted to study the changes in biochemical contents such as carbohydrates, proteins, lipids, and energy levels after 96-hour exposure to Vibrio parahaemolyticus, in flower shrimp, Penaeus semisulcatus, post-larvae. The sampling was done at intervals of 24-hours, 48-hours, 72-hours, and 96-hours. The caloric concentrations as well as biochemical constituents except protein showed a decrease in infected post-larvae. In contrast, protein showed an increase in infected post-larvae. A decrease in the ratios of carbohydrate/protein and carbohydrate/lipid was observed in infected post-larvae. The differential responses of biochemical constituents might be due to triggering of compensatory mechanism under stress conditions due to V. parahaemolyticusinfection.  相似文献   

14.
This study investigated potential application of lactic acid bacteria (LAB) in depuration for reducing Vibrio parahaemolyticus in oysters. Lactobacillus plantarum ATCC 8014, which exhibited strong bactericidal effects against V. parahaemolyticus in vitro, was added to artificial seawater for depuration of Pacific oysters (Crassostrea gigas) inoculated with V. parahaemolyticus BE 98-2029 (O3:K6) to levels of about 104 MPN/g at 15 ± 1 and 10 ± 1°C. Application of L. plantarum ATCC 8014 treatment (107 CFU/mL) in oyster depuration did not enhance reductions of V. parahaemolyticus in oysters depurated at 15 ± 1°C but significantly decreased (p < 0.05) levels of V. parahaemolyticus in oysters depurated at 10 ± 1°C after 5 days (3.40 log reductions) when compared with controls (2.75 log reductions). It is not clear if a competitive exclusion by LABs to compete with V. parahaemolyticus binding sites in oyster tissues plays a role in the reduction of V. parahaemolyticus in the oysters. Further studies utilizing different types of LABs in oyster depuration might provide additional knowledge for application of LAB in depuration for decontaminating V. parahaemolyticus in oysters.  相似文献   

15.
Vibriosis outbreaks due to Vibrio ordalii occur globally, but Chilean salmon aquaculture, in particular, has suffered significant monetary losses in the last 15 years. Little is known about the virulence mechanisms employed by V. ordalii. However, most Vibrio pathogens (e.g., Vibrio anguillarum, a very close taxonomic species) present outer membrane vesicles (OMVs) that are released extracellularly and implicated in the delivery of virulence factors to host cells. This study provides the first reported evidence of the fish pathogen V. ordalii producing and releasing OMVs under normal growth conditions. Analyses were conducted with the V. ordalii strain Vo-LM-18 and the type strain ATCC 33509T. For comparative purposes, the reference strain V. anguillarum ATCC 43307 was employed. The average size for the three Vibrio strains was 0.215 ± 0.6 µm (via scanning electron microscopy) or between 0.19 and 1.8 µm (via dynamic light scattering), with each bacterium presenting a wide range. SDS-PAGE revealed similarities in OMV patterns, but neither total nor external proteins were identical. Comparing V. ordalii ATCC 33509T and Vo-LM-18, bands were most evident in the total proteins, and the greatest degree of similarity in OMV profiles was between 37 and 50 kDa. The purified OMVs demonstrated haemolytic enzyme activity, which could play a role during V. ordalii infection. These data represent an initial step towards gaining new insights into this virulence factor, of which a lot is known in other pathogenic microorganisms.  相似文献   

16.
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17.
Vibrio parahaemolyticus, an important pathogen of freshwater prawn, Macrobrachium rosenbergii causes high mortality. Four strains of V. parahaemolyticus were isolated from the gill, haepatopancreas, hemolymph and carapace of diseased prawn. The species were characterized based on biochemical and serological tests. On performing an antibiotic assay, these strains were found to be highly susceptible to cephalaxin, tetracycline and erythomycin. During the in vitro pathogenicity test, all the strains were found to be positive to the Congo red binding assay and to be haemolytic in nature, whereas an in vivo pathogenicity test revealed that 2 × 109 CFUs mL?1of bacteria induced disease symptoms such as black colouration on the carapace, red discolouration of the exoskeleton and loss of appendages within 6 days and 80% mortality. In histopathological studies, a prominent necrosis was seen in gill lamellae, and branchial arches were thickened at places due to hyperplasia and haemocytic infiltration. Hepatopancreatic tissue showed dilation of tubules, vacuolation of hepatocytes and marked necrosis in acinar cells. Development of an immunological technique for the detection and screening of V. parahaemolyticus infection and its treatment is highly important in freshwater prawn aquaculture.  相似文献   

18.
Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.  相似文献   

19.
ABSTRACT

The present study was aimed to evaluate the effectiveness of administration of honey as a prebiotic in improving growth performance, immune response, and resistance of Pacific white shrimp Litopenaeus vannameito to Vibrio parahaemolyticus infection. In the present study, the bee honey added to practical diet at levels of 0.0%, 0.25%, 0.5%, 0.75%, and 1.0%. Shrimps were fed one of the tested diets in triplicates for 70 days. After the feeding trial, challenged was conducted with V. parahaemolyticus for two and five days. The growth performance and immune responses of shrimp in treatments with prebiotic honey were better than control one. Pacific white shrimp fed 0.75% honey showed highest survival after bacterial infection (73.33%), while lowest survival was found in the control treatment (43.33%). It may be concluded that the administration of honey at level of 0.75% was effective in improving the growth performance, immune response, and resistance of shrimp to V. parahaemolyticus infection.  相似文献   

20.
The application of probiotics in food is now widespread and is widely accepted by consumers. Lactic Acid Bacteria (LAB) were isolated from traditional salted fish and then characterized by its ability to inhibit Vibrio parahaemolyticus growth using minimal inhibitory concentration tests. Five out of these strains were identified as Lactobacillus delbrueckii, Lactobacillus casei and three Lactobacillus gasseri by PCR using 16S and 23S rRNA genes. Antibiofilm activity of Lactobacillus spp. extracts were also tested in 96 polystyrene plates. A potential antibiofilm effect was demonstrated as most LAB. Although most LAB extract were able to eradicate pre‐formed biofilm, results demonstrated that five Lactobacillus spp. exhibited a stronger inhibitory effect against V. parahaemolyticus in infected oysters. Vibrio parahaemolyticus viable cells number declined from 106 UFC to 104 UFC after 3 days of incubation with Lactobacillus spp. Probiotic applications, in biological control of seafood associated pathogens can be an alternative solution, providing consumer with a product of good quality owing to the use of non‐toxic compounds. Based on our results, LAB could be used as a bioprotective culture in oyster's depuration to prevent V. parahaemolyticus growth.  相似文献   

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