Antimicrobial resistance survey in a river receiving effluents from freshwater fish farms

AIMS: The aims of this study were: (i) to determine the proportions of Aeromonas spp. resistant to florfenicol (FC), oxolinic acid (OA) and oxytetracycline (OTC) along a river receiving effluents from fish farms, and (ii) to assess the relevance of using this bacterial group as an indicator for studying the consequences of the use and release of these aquacultural antimicrobials in the freshwater environment, as compared with performing antimicrobial measurements in sediments. METHODS AND RESULTS: Sediment interstitial waters sampled along a river during two distinct climatic seasons were plated on an Aeromonas-selective medium supplemented or not with OA, OTC or FC. The October 2004 campaign showed an enrichment of OA- and OTC-resistant Aeromonas immediately downstream of the fish farms and a wastewater treatment plant. Two fish farms showed similar results in March 2005. In contrast, only 10 FC-resistant Aeromonas strains could be isolated, which revealed that minimum inhibitory concentrations of FC were greater than 64 microg ml(-1) and multiple antimicrobial resistances. Contamination of sediments by antimicrobials was detected but was not always co-localized with resistance peaks or known point sources of contamination. CONCLUSIONS: Aeromonas could be valuable indicators of OA, OTC and FC resistance in the freshwater environment. Fish farms contribute to the contamination of the river by antimicrobials and resistant bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Considering the still very low proportion of FC-resistant Aeromonas, this study can be considered as a reference for further studies about this recently introduced veterinary antimicrobial agent.     

In vitro antibiotic susceptibility of bacteria isolated from EUS-affected fishes in India

AIMS: Twelve antibiotics were evaluated for in vitro sensitivity against 16 bacterial strains isolated from surface lesions of fishes affected with epizootic ulcerative syndrome (EUS). METHODS AND RESULTS: Disc diffusion assay in Mueller-Hinton agar showed that the pseudomonads and aeromonads were mainly resistant to penicillin, ampicillin and erythromycin. Additionally, some were resistant to gentamycin and amoxycillin. However, resistance towards antibiotics previously recommended for EUS treatment, such as oxytetracycline and chloramphenicol, was not observed. Four aeromonads and two pseudomonads were found to induce ulcers when injected intramuscularly in healthy Anabas testudineus. CONCLUSIONS: All six pathogenic isolates were sensitive towards oxytetracycline, chloramphenicol and nalidixic acid. Oxytetracycline seems to be an effective antibiotic, and further investigations to determine the mode of treatment and dose appear to be worthwhile.     

Antimicrobial resistance in bacteria isolated from aquaculture sources in Australia

AIMS: To carry out a preliminary assessment of the occurrence of resistance to antimicrobials in bacteria that has been isolated from a variety of aquaculture species and environments in Australia. METHOD AND RESULTS: A total of 100 Gram-negative (Vibrio spp. and Aeromonas spp. predominantly) and four Gram-positive bacteria isolated from farmed fish, crustaceans and water from crab larval rearing tanks were obtained from diagnostic laboratories from different parts of Australia. All the isolates were tested for sensitivity to 19 antibiotics and Minimal Inhibitory Concentrations were determined by the agar dilution method. Plasmid DNA was isolated by the alkali lysis method. Resistance to ampicillin, amoxycillin, cephalexin and erythromycin was widespread; resistance to oxytetracycline, tetracycline, nalidixic acid and sulfonamides was common but resistance to chloramphenicol, florfenicol, ceftiofur, cephalothin, cefoperazone, oxolinic acid, gentamicin, kanamycin and trimethoprim was less common. All strains were susceptible to ciprofloxacin. Multiple resistance was also observed and 74.4% of resistant isolates had between one and ten plasmids with sizes ranging 2-51 kbp. CONCLUSIONS: No antibiotics are registered for use in aquaculture in Australia but these results suggest that there has been significant off-label use. SIGNIFICANCE AND IMPACT OF STUDY: Transfer of antibiotic resistant bacteria to humans via the food chain is a significant health concern. In comparison with studies on terrestrial food producing animals, there are fewer studies on antibiotic resistance in bacteria from aquaculture enterprises and this study provides further support to the view that there is the risk of transfer of resistant bacteria to humans from consumption of aquaculture products. From the Australian perspective, although there are no products registered for use in aquaculture, antimicrobial resistance is present in isolates from aquaculture and aquaculture environments.     

不同来源株香港海鸥形菌药敏试验结果分析

目的对分离自社区获得性胃肠炎患者及淡水鱼肠道内的香港海鸥形菌(Laribacter hongkongensis,LH)进行药敏试验,为临床合理使用抗菌药物提供依据。方法2005年至2006年杭州市第一人民医院肠道门诊从社区获得性胃肠炎患者中分离到4株及2种淡水鱼体内10株LH菌株,计14株,按美国CLSI推荐的纸片扩散法进行药敏试验。结果药敏结果显示,14株LH菌株对24种常用抗菌药物中的15种药物,分别表现出不同程度的耐药性,耐药率达62.5%;尤其对青霉素、头孢唑啉和头孢哌酮等3种药物,14株LH菌株均为耐药,耐药率为100.0%;其中人体株LHHZ26,对12种药物耐药、1种为中介。LH对3代头孢表现为较高的耐药性,3种头孢药物的耐药率为71.43%~100%;14株LH株对头孢吡肟及头孢哌酮/舒巴坦均为敏感。对氨曲南(14.29%)、庆大霉素(7.14%)、复方新诺明(7.14%)的耐药率较低;喹诺酮类、氯霉素、阿米卡星、美罗培南、亚胺培南等药物均敏感。结论LH菌株对β-内酰胺类药物表现出耐药性,对青霉素类、头孢菌素类药物广泛耐药。建议对LH感染患者喹诺酮类为首选治疗用药。     

Antimicrobial resistance in Gram-negative bacilli isolated from infant formulas

A total of 90 samples of infant formula (IF) were collected from the lactary of a teaching hospital, during a 4-month period from July to August 1999. The sanitary conditions of the formulas were analyzed, and a physiological characterization of Gram-negative bacillus isolates and antimicrobial susceptibility testing were performed. Colony counts were considered to be unacceptable for the majority of the IF samples and the contamination rates were related to inadequate handling. Coliforms (35 degrees C and 45 degrees C growth) were detected in most of the IF tested. Klebsiella pneumoniae, Citrobacter freundii, Cedacea davisae, Klebsiella planticola and Enterobacter cloacae were the isolates most commonly identified. Antimicrobial susceptibility testing showed significant resistance rates, particularly to amoxicillin/clavulanic acid, cefoxitin, cephalotin or ampicillin. One extended-spectrum beta-lactamase-producing K. pneumoniae strain was also recovered.     

新生儿重症监护病房常见病原菌耐药率比较分析

目的探讨新生儿重症监护病房与其他病房分离病原菌的耐药率。方法采用VITEK全自动微生物分析仪进行药物敏感性分析,对深圳市儿童医院1999～2004年分离的2组常见病原菌的耐药率进行统计分析,耐药率的显著性比较用χ2检验。结果新生儿重症监护病房常见病原菌为表皮葡萄球菌、凝固酶阴性葡萄球菌和肺炎克雷伯菌,而其他病房以金黄色葡萄球菌、表皮葡萄球菌、凝固酶阴性葡萄球菌、大肠埃希菌和铜绿假单胞菌为主。新生儿重症监护病房病原菌对左氧沙星、环丙沙星、头孢吡肟、头孢他啶和哌拉西林三唑巴坦的耐药率分别为376%、368%、712%、712%和505%,显著高于儿科其他病房分离菌株的耐药率(P<0001)。结论新生儿重症监护病房分离菌的耐药率高于其他病房病原菌的耐药率。     

Incidence of haemolysin-positive and drug-resistant Aeromonas hydrophila in freshly caught finfish and prawn collected from major commercial fishes of coastal South India

The incidence of Aeromonas hydrophila in freshly caught finfish and prawns from four major commercial fish landing sites of coastal South India was studied for a period of one year. Among 514 analysed samples of seafood (410 finfish and 104 prawn), 37% of them (37.3% of finfish and 35.6% of prawn) were contaminated with A. hydrophila. A total of 255 strains of A. hydrophila were isolated. Of the total isolates, about 78.4% of them were producers of haemolysin. All strains were resistant to bacitracin and all were sensitive to chloramphenicol. The results indicate that the strains originated from high-risk sources. The presence of A. hydrophila is an indication of marine contamination. The increasing presence of haemolysin-producing multiple drug-resistant A. hydrophila in fish and prawn may become a potential human health hazard.     

Antimicrobial resistance of bacteria isolated from slaughtered and retail chickens in South Africa

Animal feed is increasingly being supplemented with antibiotics to decrease the risk of epidemics in animal husbandry. This practice could lead to the selection for antibiotic resistant micro-organisms. The aim of this study was to determine the level of antibiotic resistant bacteria present on retail and abattoir chicken. Staphylococci, Enterobacteriaceae, Salmonella and isolates from total aerobic plate count were tested for resistance to vancomycin, streptomycin, methicillin, tetracycline and gentamicin using the disc diffusion susceptibility test; resistance to penicillin was determined using oxacillin. Results from the antibiotic code profile indicated that many of the bacterial strains were displaying multiple antibiotic resistance (MAR). A larger proportion of resistance to most antibiotics, except for vancomycin, was displayed by the abattoir samples, therefore suggesting that the incidence of MAR pathogenic bacteria was also higher in the abattoir samples. This resistance spectrum of abattoir samples is a result of farmers adding low doses of antibiotics to livestock feed to improve feeding efficiency so that the animals need less food to reach marketable weight. The lower incidence of MAR pathogenic bacteria in the retail samples is a result of resistance genes being lost due to lack of selective pressure, or to the fact that the resistant flora are being replaced by more sensitive flora during processing. The use of subtherapeutic levels of antibiotics for prophylaxis and as growth promoters remains a concern as the laws of evolution dictate that microbes will eventually develop resistance to practically any antibiotic. Selective pressure exerted by widespread antimicrobial use is therefore the driving force in the development of antibiotic resistance. This study indicated that a large proportion of the bacterial flora on fresh chicken is resistant to a variety of antibiotics, and that resultant food-related infections will be more difficult to treat.     

Antimicrobial resistance in Campylobacter isolated from food animals and humans in northern Thailand

A study was conducted to determine the prevalence of Campylobacter with antimicrobial resistance from chickens, pigs, dairy cows, healthy farm workers, and children hospitalized with diarrhea in northern Thailand. Resistance was highest in pig samples and lowest in healthy farm workers. Resistance to fluoroquinolones and tetracycline was high in all study populations. The increased prevalence of resistant isolates from meat samples collected at markets, compared to isolates collected from animals on the farm or the slaughterhouse, suggests that contamination after carcasses leave the slaughterhouse is an important factor in the spread of resistant bacteria into the human food chain.     

Screening of benzalkonium chloride resistance in Listeria monocytogenes strains isolated during cold smoked fish production

AIMS: To determine the susceptibility to disinfectants and cross-resistance to antibiotics in Listeria monocytogenes strains isolated from fish products and the fish-processing environment. METHODS AND RESULTS: Minimal inhibitory concentration assessment, using the agar dilution method, showed 108 of 255 L. monocytogenes isolates with low susceptibility to benzalkonium chloride (BC), commonly used in food industries. Most of them are from raw products of farmed fish during processing, while the remaining resistant isolates were mainly from the environment and finished products irrespective of the fish species. Two BC-resistant isolates were resistant to ethidium bromide (EB). The conservation of resistance after plasmid curing suggested that the resistance genes are not plasmid associated. EB accumulation assays demonstrated that the two BC(R) EB(R) isolates used an efflux pump to expel these substrates whereas a different mechanism was probably used by the majority of the strains with BC(R) EB(S) pattern. No cross-resistance was found with antibiotics. CONCLUSIONS: This study highlights the difference in susceptibilities to BC for L. monocytogenes strains isolated from fish-processing plants and in resistance mechanisms to BC developed by these bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of BC resistant L. monocytogenes strains could contribute to their adaptation and so explained their survival and persistence in the fish-processing environment.     

Occurrence of foodborne pathogenic bacteria in retail prepackaged portions of marine fish in Spain

AIMS: To survey the presence of indigenous and nonindigenous foodborne bacterial pathogens in displayed prepacked portions of fresh marine fish. METHODS AND RESULTS: A survey of 50 different samples of fresh marine fish (conger, swordfish, sole, grouper and whiting) was conducted over a period of 5 months. Trays of fillets and steaks were obtained at retail level and tested for foodborne bacterial pathogens. Vibrio cholerae and Salmonella were not detected. Two samples (4%) yielded Vibrio strains carrying a DNA fragment specific for Vibrio parahaemolyticus, but resulted negative to PCR amplification of the virulence-related tdh gene. Levels of motile Aeromonas ranging from 2.29 to 7.20 log CFU g(-1) were found in 31 (62%) samples. All fish portions were positive for the Aeromonas hlyA gene and 38 for both aerA and hlyA genes, which may contribute to diarrhoea-related virulence. The incidence of Listeria monocytogenes was 10%. Levels of Staphylococcus aureus lower than 2 log CFU g(-1) were found in 15 (30%) samples. Numbers of presumptive Clostridium perfringens ranging from 1.82 +/- 0.22 to 4.26 +/- 1.25 log CFU g(-1) were detected in 42 (84%) samples. Edwardsiella tarda was detected in two samples of grouper fillets. CONCLUSIONS: Displayed portions of raw fish carried bacteria that can cause foodborne disease. The risk posed by fresh fish when properly cooked is low, but high when destined to be consumed raw, undercooked or very lightly processed. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed that raw fish sold in Spain could be a source of foodborne bacterial pathogens. Improvements in handling and processing are needed to minimize the prevalence of pathogenic bacteria.     

Antimicrobial resistance and biotechnological potential of plastic-associated bacteria isolated from an urban estuary

Plastics have quickly become one of the major pollutants in aquatic environments worldwide and solving the plastic pollution crisis is considered a central goal of modern society. In this study, 10 different plastic samples, including high- and low-density polyethylene and polypropylene, were collected from a deeply polluted urban estuary in Brazil. By employing different isolation and analysis approaches to investigate plastic-associated bacteria, a predominance of potentially pathogenic bacteria such as Acinetobacter, Aeromonas, and Vibrio was observed throughout all plastic samples. Bacteria typically found in the aquatic environment harboured clinically relevant genes encoding resistance to carbapenems (bla_KPC) and colistin (such as mcr-3 and mcr-4), along with genetic determinants associated with potentially active gene mobilization. Whole genome sequencing and annotation of three plastic-associated Vibrio strains further demonstrated the carriage of mobile genetic elements and antimicrobial resistance and virulence genes. On the other hand, bacteria isolated from the same samples were also able to produce esterases, lipases, and bioemulsifiers, thus highlighting that the plastisphere could also be of special interest from a biotechnological perspective.     

胆汁分离细菌种类及主要分离菌株耐药性分析

目的了解2000年至2008年临床胆汁普通培养分离细菌的种类,并分析主要分离菌株的耐药性。方法回顾性分析近9年710例胆囊炎和胆囊结石患者胆汁普通培养分离细菌的分布情况及耐药率;用微量稀释法进行药物敏感性测定。结果710例患者胆汁培养共分离出细菌435株,检出率为61.27%,前5位分别为大肠埃希菌、肠球菌、假单胞菌、肺炎克雷伯菌和凝固酶阴性葡萄球菌。革兰阴性杆菌对亚胺培南、阿米卡星的耐药率均低于10%,对常用青霉素类和头孢菌素类抗生素耐药率超过50%。结论胆汁分离细菌以肠道细菌为主,阿米卡星可以作为经验治疗的首选药物。