Phenotypic characterization and antibiotic susceptibility of Vibrio spp. isolated from aquaculture waters on the west coast of Sabah, Malaysia
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The phenotypic characterization and antibiotic susceptibility patterns of 72 environmental isolates of Vibrio spp. collected from six aquaculture sites along the west coast of Sabah, Borneo, Malaysia was investigated. Up to 21 conventional biochemical tests were carried out and the sensitivity of the isolates against 16 antibiotics were tested using the disk diffusion method. In the present study, 11 species of vibrios were isolated with varying percentage of occurrences, with the three most abundant species being Vibrio harveyi (22.2%), V. parahaemolyticus (22.2%) and V. alginolyticus (19.4%). The isolates produced varying results in the biochemical tests. All environmental Vibrio isolates were found sensitive to chloramphenicol, nalidixic acid and oxolinic acid but exhibited resistance to vancomycin and penicillin. Susceptibility was variable for other antibiotics. As for V. alginolyticus, it was found that 100% of the isolates were sensitive to chloramphenicol, nalidixic acid and oxolinic acid but resistant to ampicillin, novobiocin, penicillin and vancomycin. Meanwhile, all V. harveyi isolates were sensitive to chloramphenicol, furazolidone, nalidixic acid,nitrofurantoin and oxolinic acid but resistant to penicillin and vancomycin. All V. parahaemolyticus isolates were observed sensitive to chloramphenicol, nalidixic acid and oxolinic acid but resistant to ampicillin, penicillin and vancomycin. Since all three are important species implicated in fish and shrimp disease, and two (V. parahaemolyticus and V. alginolyticus) can cause clinical ailments in humans, microbial diversity at similar sites should be investigated further.Keywords:
Oxolinic acid
Vibrio alginolyticus
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Antimicrobial susceptibility patterns and plasmid profiles of Vibrio species isolated from edible crabs ( Callinectes sapidus ) were evaluated. A total of 40 crabs and the obtained water samples collected from four different markets in Lagos State were examined. Morphological and biochemical techniques were used to identify the Vibrio spp present in the samples. Susceptibility of bacteria isolates to 8 antibiotics was studied using a disc diffusion method and plasmid profile was determined by agarose gel electrophoresis. The total viable Vibrio count in unprocessed samples ranged from 3.30 to 5.80 Log10cfu/g while the processed ones was 1.80 to 2.5 Log10cfu/g. The identified Vibrio spp included, V. cholerae, V. harveyi. V. mimicus, V. parahaemolyticus, V. furnissii and V. fluvalis . Results revealed that Vibrio isolates from the samples expressed total (100%) antibiotic resistance to amoxcycilin and augumentin, 83.3% to cotrimoxazole, 72.2% to tetracycline, 66.6% to nitrofurantoin, 61.6% to gentamicin, 33.3% to nalidilixic acid and no resistance to ofloxacin. The isolates had a multiple antibiotic resistance (MAR) index ranging from 0.25–0.88. Plasmid profile of 10 isolates which showed multiple resistance to antibiotics revealed that 7 harboured plasmid with bands ranging from 23130 bp – 20240 bp as molecular weight. Isolation of Vibrio spp from seafood of this nature is considered as a potential danger for consumers.
Callinectes
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One thousand forty-nine bacterial isolates, which were collected from shrimp farms in Thailand, were resistant to antimicrobial agents: ampicillin (ABPC), chloramphenicol (CP), kanamycin (KM), oxolinic acid (OA), streptomycin (SM) and tetracycline (TC). One hundred forty-two isolates were identified as Vibrio. These isolates were conjugated with Escherichia coli for detection of transferable R plasmid. Six patterns of transferable R plasmids, which encoded resistance to the combinations of one to five drugs (ABPC, CP, SM, sulfamonomethoxine, TC and trimethoprim), were found from 16 drug-resistant isolates of Vibrio. This is the first report of transferable R plasmids from Vibrio spp. in shrimp farms in Thailand.
Kanamycin
Oxolinic acid
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Vibrio alginolyticus
Kanamycin
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Sulfamethoxazole
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This study was performed to investigate the presence of halophylic Vibrio spp. in horse-mackerel (Trachurus trachurus L., 1758). In total, 80 fish samples were collected from fish markets and supermarkets between 04.04.2006 and 16.05.2006 in the city of Mersin. All fish samples were microbiologically tested for halophylic Vibrio spp. Nine (11.25%) Vibrio spp. strains were isolated and identified as V. alginolyticus. Molecular epidemiological discrimination and antimicrobial susceptibility patterns of V. alginolyticus strains were also analysed by arbitrarily primed polymerase chain reaction (AP-PCR) and Kirby-Bauer disc diffusion, respectively. Genotypic heterogeneity of these 9 isolates discriminated by AP-PCR determined that while only 3 isolates were from the same genotypic source the other 6 isolates were from different sources. All isolates were resistant to ampicillin, cephalothin, and streptomycin, and susceptible to levofloxacin, ofloxacin, cefotaxime, nalidixic acid, tetracycline, piperacillin-tazobactam, chloramphenicol, imipenem, and aztreonam. One isolate was resistant to cefepime, 1 to nitrofurantoin, and 1 to ciprofloxacin. One of the 9 isolates was susceptible to erythromycin and 1 to novobiocin, and 3 isolates were susceptible to sulbactam-cefoperazone.
Vibrio alginolyticus
Cefoperazone
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A total of 51 potential pathogenic vibrios were isolated from moribund silver seabream Sparus sarba, which were collected from local fish farms of Hong Kong. All the isolates were classified and identified as 7 species by the API 20 E system and the scheme of Alsina & Blanch. These species were Vibrio alginolyticus (24 strains), Vibrio vulnificus (12 strains), Vibrio parahaemolyticus(7 strains), Vibrio logei(4 strains), Vibrio pelagius II(2 strains), Vibrio fluvialis (1 strains) and Vibrio meditterranei (1 strains). Among these isolates, the three predominant species (V. alginolyticus, V. vulnificus and V. parahaemolyticus) were confirmed to be virulent to sea bream by experimental challenge. All isolates were also screened for plasmid DNA by agarose gel electrophoresis and tested for susceptibility to 16 antimicrobial agents by the agar dilution method. Of the 51 isolates examined, all strains were sensitive to ceftriaxone, streptomycin, nalidixic acid and rifampicin, and almost all were sensitive to ceftazidime, netilimicin, chloramphenicol and sulfamethoxazole except one or two strains. Most isolates were resistant to ampicillin (60. 8%), cefuroxime (66.7%), amikacin(55%), kanamycin(58.8%) and trimethoprinm (76.5%). Fifteen of the 51 isolates harboured 1-4 plasmids, with sizes ranging from 9 to 123 kb. Both the plasmids and the associated antimicrobial resistance (ampicillin, cefuroxime and trimethoprim) of 9 isolates could be transferred to recipient by single-step conjugation, however, the frequencies were very low, ranging from 10(-11) to 10(-9). The present results indicate that resistance to these antibiotics is chromosomal.
Vibrio alginolyticus
Vibrio vulnificus
Kanamycin
Amikacin
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Vibrio parahaemolyticus is a widespread bacterium in the marine environment and is one of the leading causes of food-derived bacterial poisoning in humans worldwide. The main objective of this study was to determine the prevalence, virulence factors, and antibiotic and heavy metal resistance profiles of V. parahaemolyticus in Pacific mackerel (Pneumatophorus japonicus) from different markets in Zhejiang Province, People's Republic of China. In total, 112 (31.11%) V. parahaemolyticus isolates were identified from 360 Pacific mackerel samples, with an extremely low occurrence of the virulence genes trh (1.79%, 2 of 112) and tdh (0%, 0 of 112). Antibiotic resistance testing revealed that most isolates showed resistance to ampicillin (96.43%, 108 of 112) and streptomycin (90.18%, 101 of 112), whereas all strains were sensitive to kanamycin, florfenicol, cefamandole, and trimethoprim-sulfamethoxazole. Furthermore, 46.43% (52 of 112) of isolates, which had 12 different phenotypes, were classified as multidrug resistant. In addition, the multiple antibiotic resistance index values of isolates were between 0.05 and 0.63, and the maximum multiple antibiotic resistance index was attributed to two isolates that exhibited resistance to 12 antibiotics. Heavy metal resistance patterns were similar among the six different markets. The majority of isolates showed resistance to Cd2+ (78.57%) and Pb2+ (51.79%), and fewer were resistant to Cu2+ (37.50%), Zn2+ (25.00%), Co2+ (9.82%), Ni2+ (6.25%), and Mn2+ (4.46%). No isolates were resistant to Cr3+. In total, 22.32% (25 of 112) of strains were multiheavy metal resistant. Furthermore, multidrug resistance and multiheavy metal resistance were found to be positively correlated in the V. parahaemolyticus strains by using Pearson's correlation analysis (P = 0.008; R = 0.925). This information will contribute to the monitoring of variations in the antibiotic and heavy metal resistance profiles of V. parahaemolyticus strains from seafood and provide insight into the appropriate use of antibiotics and the safe consumption of seafood.
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Abstract Background Numerous prevalence studies of Vibrio spp. infection in fish have been extensively reported worldwide, including Malaysia. Unfortunately, information on the prevalence of Vibrio spp. in groupers ( Epinephelus spp.) is limited. In this study, groupers obtained from nine farms located at different geographical regions in Malaysia were sampled for the presence of pathogenic Vibrio spp. and their susceptibility profiles against seven antibiotics. Results Out of 270 grouper samples, 195 (72%) were detected with the presence of Vibrio spp. Vibrio communis showed highest prevalence in grouper (28%), followed by V. parahaemolyticus (25%), V. alginolyticus (19%), V. vulnificus (14%), V. rotiferianus (3%), Vibrio sp. (3%), V. campbellii (2%) , V. mytili (2%) , V. furnissii (2%) , V. harveyi (1%) , V. tubiashii (1%) , V. fluvialis (0.3%) and V. diabolicus (0.3%) . Assessment on the antibiotic susceptibility profiles of the Vibrio spp. revealed that majority of the isolates were susceptible to tetracycline, streptomycin, erythromycin and bacitracin, but resistance to ampicillin, penicillin G and vancomycin. The mean MAR index of the Vibrio isolates was 0.51, with 85% of the isolates showed MAR index value of higher than 0.2. Results indicate that the Vibrio spp. were continuously exposed to antibiotics. Furthermore, the plasmid profiles of Vibrio spp. showed that 38.7% of the isolates harbored plasmid with molecular weight of more than 10 kb, while 61.3% were without plasmid. During curing process, Vibrio spp. lost their plasmid, but remained resistant to ampicillin, penicillin G, bacitracin and vancomycin while a few isolates remained resistant to erythromycin, streptomycin and tetracycline. The results suggested that the resistance to antibiotics in isolated Vibrio spp. might be due to chromosomal and plasmid borne. Conclusions This study demonstrates the prevalence of Vibrio spp. in groupers and the distribution of multidrug resistance strains that could be of concern to the farmers in Malaysia. In addition, data from this study can be further used in fish disease management plan.
Vibrio alginolyticus
Bacitracin
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