Antimicrobial resistance patterns in Enterobacteriaceae isolated from an urban wastewater treatment plant

Over 18 months, enterobacteria were isolated from the raw (189 isolates) and treated (156 isolates) wastewater of a municipal treatment plant. The isolates were identified as members of the genera Escherichia (76%), Shigella (7%), Klebsiella (12%) and Acinetobacter (4%). Antimicrobial susceptibility phenotypes were determined using the agar diffusion method for the antibiotics amoxicillin, gentamicin, ciprofloxacin, sulfamethoxazole/trimethoprim, tetracycline and cephalothin, the disinfectants hydrogen peroxide, sodium hypochlorite, quaternary ammonium/formaldehyde and iodine, and the heavy metals nickel, cadmium, chromium, mercury and zinc. Class 1 integrons were detected by PCR amplification using the primers CS5 and CS3. Compared with the raw influent, the treated wastewater presented higher relative proportions of Escherichia spp. isolates resistant to ciprofloxacin and cephalothin (P<0.0001 and P<0.05, respectively). Except for mercury, which showed a positive correlation with tetracycline and sulfamethoxazole/trimethoprim, no significant positive correlations were observed between antibiotic, disinfectant and heavy metal resistance. The variable regions of class 1 integrons, detected in c. 10% of the Escherichia spp. isolates, contained predominantly the gene cassettes aadA1/dhfrI.     

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.     

Antibiotic resistance among coliform bacteria isolated from hospital and urban wastewaters

The mean count of coliforms/ml in urban and hospital drains were 68.9×10⁴ and 48.4×10⁴, respectively.Klebsiella predominated amongst the 16 coliform species that were differentiated;Escherichia coli occurred with highest frequency. All the isolates were resistant to at least one antibiotic and resistance to ampicillin, tetracycline, and chloramphenicol was particularly high. Multiple antibiotic resistant strains were common in both environments—85% and 65% for hospital and urban sources, respectively. The difference in the frequency of resistant strains from the two sources was not significant. 20% of strains from the two sources were multiply resistant to six antibiotics. The role of untreated wastewater in the maintenance and dissemination of resistant coliforms and its public health significance are highlighted.

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Résumé Le comptage moyen de coliformes par ml dans des égouts urbains ou d'hôpital était respectivement de 68.9 10⁴ et de 48.4 10⁴.Kiebsiella prédominait parmi les 16 espèces de coliformes différenciées;Escherichia coli apparaissait avec la fréquence la plus élevée. Tours les isolats étaient résistants à au moins un antibiotique et la résistance à l'ampicilline, la tetracycline et le chloramphénicol était particulièrement élevée. Des souches résistantes à plusieurs antibiotiques étaient communes dans les deux environnements—85% et 65% respectivement pour les origines urbaines et d'hôpital. La différence dans la fréquence de souches résistantes provenant des deux origines n'était pas significative. Vingt pour-cent des souches provenant des deux origines présentaient une résistance multiple aux six antibiotiques. On a mis en évidence le rôle de l'eau résiduaire non traitée dans le maintien et la dissémination de coliformes résistants et de sa signification pour la santé publique.

     

Numerical taxonomy of heavy metal-tolerant bacteria isolated from an estuary.

A total of 230 strains of metal-tolerant bacteria from water and sediment samples collected in Chesapeake Bay were isolated on a medium containing cobalt, lead, mercury, or molybdenum. In addition, a set of 71 cultures were simultaneously isolated on glucose tryptone yeast extract agar medium without metals. Twenty-three reference strains were also included in the numerical taxonomy study of these bacteria, bringing the grand total of strains examined to 324. All strains were examined for 112 biochemical, cultural, morphological, and physiological characters. The taxonomic data obtained were analyzed by computer and the simple matching (SSM) and Jaccard (SJ) coefficients were calculated. Clustering achieved by unweighted average linkage is presented and, from sorted similarity matrices and dendrograms, 294 strains, i.e., 97% of the total, were recovered in 12 phenetic groups defined at the 75 to 80% similarity level. Among the strains there were nine phena presumptively identified as Bacillus, Erwinia, Mycobacterium, Pseudomonas, and coryneforms. From the results of the taxonomic study, it is concluded that metal tolerance in estuarine water and sediment bacteria occurs among a restricted range of taxa distributed throughout the estuarine environment.     

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.     

Ecology and taxonomy of chitinoclasticCytophaga and related chitin-degrading bacteria isolated from an estuary

A total of 103 strains of estuarine, Chitinoclastic bacteria isolated from water, and sediment samples collected from the upper Chesapeake Bay, including 17 freshwater and 11 seawater isolates, were subjected to numerical taxonomy analysis. The isolates included 44 yellow-orange pigmented strains classified asCytophaga-like bacteria (CLB) of theCytophagaceae. Salt requirement of the strains ranged from tolerance to 1% NaCl to an absolute requirement for NaCl, with 1% NaCl satisfying this requirement. The largest phenon consisted of facultatively anaerobic, oligo-nitrophilic, and flexirubin pigment-producing freshwater and estuarine isolates, and included reference strains of bothCytophaga johnsonae Stanier andCytophaga aquatilis Strohl and Tait. Other phena, containing a smaller number of strains, comprised marine and estuarine isolates which did not produce flexirubin pigments, and required organic nitrogen for growth and for production of chitinolytic enzymes. Salt-requiring, flexirubin pigment-producing, chitin-degrading strains were, on occasion, isolated from estuarine samples and represented phena found in estuaries. Most of theCytophaga isolates, as well as chitin-degrading species not of the genusCytophaga that were isolated from Chesapeake Bay, clustered in phena representing previously described species of aerobic, zymogenic, chitinoclastic bacteria. When the frequency of occurrence of features related to environmental parameters, viz., pH, salinity, temperature range of growth, and growth on media lacking organic nitrogen, was calculated, ecological groupings of strains in the 2 major phena of CLB could be distinguished among the estuarine, chitin-degrading bacteria.     

Diversity and biotechnological potential of culturable bacteria from Brazilian mangrove sediment

Mangrove ecosystems are environments subject to substantial degradation by anthropogenic activities. Its location, in coastal area, interfacing the continents and the oceans makes it substantially important in the prospection for biotechnological applications. In this study, we assessed the diversity of culturable bacteria present over the seasons at two depths (0–10 and 30–40 cm) in a mangrove sediment and in a transect area from the land to the sea. In total, 238 bacteria were isolated, characterized by Amplified Ribosomal DNA Restriction Analysis (ARDRA) and further identified, by Fatty Acid Methyl Esther (FAME-MIDI), into the orders of Vibrionales, Actinomycetales and Bacillales. Also the ability of the isolates in producing economically important enzymes (amylases, proteases, esterases and lipases) was evaluated and the order Vibrionales was the main enzymatic source.     

Fungi isolated from olive ecosystems and screening of their potential biotechnological use

This study investigated the fungi diversity of fresh olive (Olea europaea L.) fruits, olive paste (crushed olives) and olive pomace (solid waste) and screened and quantified enzymatic activities with biotechnological applications. Fungi were randomly isolated from olive cultivars from Castilla La Mancha region (Spain). Identification included comparison of their polymerase chain reaction (PCR) amplicons of the ITS1-5.8S-ITS2 ribosomal DNA region, followed by nucleotide sequence analysis. Fourteen different species with DNA sequences of different similarities were identified, belonging to seven different genera (Aspergillus, Penicillium, Rhizomucor, Mucor, Rhizopus, Lichtheimia and Galactomyces). Aspergillus fumigatus, followed by Galactomyces geotrichum, Penicillium commune and Rhizomucor variabilis var. regularior were the most frequent species. Specific enzyme screening was assayed on agar plates, using cellobiose, carboxymethylcellulose (CMC), polygalacturonic acid and CaCl(2)/Tween 80 as substrates for β-glucosidase, carboxymethylcellulase (CMCase), polygalacturonase and lipase, respectively. Species exhibiting the best activities were: Aspergillus fumigatus (for β-glucosidase, CMCase and lipase); Rhizopus oryzae (for β-glucosidase and lipase); Rhizomucor variabilis (for β-glucosidase, CMCase and polygalacturonase); Mucor fragilis (β-glucosidase, CMCase and lipase); Galactomyces geotrichum (for β-glucosidase, polygalacturonase and lipase) and Penicillium commune and Penicillium crustosum (for lipase). The species that had shown the best enzymatic activities were grown on hemicellulose, cellulose and pectin and some activities were quantified (xylanase, cellulase, β-glucosidase and pectinase). An isolate of A. fumigatus and one of A. niger showed the best cellulase and xylanase activities, while no species presented good pectinase and β-glucosidase activities. The selected species with potential enzymatic activities could be used for future applications of industrial interest.     

Antimicrobial resistance of Enterococcus species isolated from produce

The purpose of this study was to characterize the antibiotic resistance profiles of Enterococcus species isolated from fresh produce harvested in the southwestern United States. Among the 185 Enterococcus isolates obtained, 97 (52%) were Enterococcus faecium, 38 (21%) were Enterococcus faecalis, and 50 (27%) were other Enterococcus species. Of human clinical importance, E. faecium strains had a much higher prevalence of resistance to ciprofloxacin, tetracycline, and nitrofurantoin than E. faecalis. E. faecalis strains had a low prevalence of resistance to antibiotics used to treat E. faecalis infections of both clinical and of agricultural relevance, excluding its intrinsic resistance patterns. Thirty-four percent of the isolates had multiple-drug-resistance patterns, excluding intrinsic resistance. Data on the prevalence and types of antibiotic resistance in Enterococcus species isolated from fresh produce may be used to describe baseline antibiotic susceptibility profiles associated with Enterococcus spp. isolated from the environment. The data collected may also help elucidate the role of foods in the transmission of antibiotic-resistant strains to human populations.     

Antimicrobial peptides isolated from insects and their potential applications

Antimicrobial peptides (AMPs) in insects have the potential to be developed as chemotherapy agents against numerous microbial species. This article reviewed the existing knowledge of what have been focused so far on published materials related to AMPs isolated from insects. Previous studies were focused on peptide characterization and the mechanism pathways of different AMPs from a variety of insect Orders. Most studied insect Orders are as follows: Hymenoptera (50%), Diptera (17%), Coleoptera (13%), Lepidoptera (10%), Hemiptera (5%), Blattodea (3%) and Odonata (2%). Dozens of new AMPs have been extracted from insects recently. However, more studies in vivo and in vitro are necessary to fully understand their effect and the mechanisms of antimicrobial action to utilize their promising potential in cosmetic and pharmaceutical industries.     

尿液分离鲍曼不动杆菌耐药性及喹诺酮耐药机制研究

目的研究武警浙江省总队医院尿液分离鲍曼不动杆菌患者的临床特点、细菌的耐药性及菌株对喹诺酮的耐药机制。方法收集2011年1月至2011年12月临床尿液标本中分离的鲍曼不动杆菌,回顾性分析患者的临床特点,分析其对临床常用抗菌药物的耐药性,并使用PCR方法筛查qnr A、qnr B、qnr S、qep A、aac(6')-Ib-cr,gyr A和par C基因,并对阳性结果进行基因测序分析。结果共收集尿液分离鲍曼不动杆菌53株,均分离自住院患者,其中重症监护病房和外科病房患者占88.7%、留置导尿患者占94.3%,拨除导尿管并给予适当抗菌治疗后临床预后良好。菌株对临床常见抗菌药物耐药率较高,其中头孢哌酮/舒巴坦较低为15.1%、米诺环素为20.8%、阿米卡星为22.6%、美罗培南为26.4%、亚胺培南为28.3%、氨苄西林/舒巴坦为32.1%,其余药物耐药率超过40%。25株环丙沙星耐药菌株中有17株gyr A基因突变,12株par C基因突变,1株qnr B基因阳性,未发现qep A阳性菌株,7株aac(6')-Ib阳性,经序列分析确定未发生aac(6')-Ib-cr突变。结论尿液分离鲍曼不动杆菌主要来自于院内接受留置导尿的患者,其对抗菌药物耐药率较高,合适的治疗临床预后良好。对喹诺酮耐药机制主要为gyr A和par C基因突变,并出现了qnr B基因。     

Consortia of cyanobacteria/microalgae and bacteria: biotechnological potential

Microbial metabolites are of huge biotechnological potential and their production can be coupled with detoxification of environmental pollutants and wastewater treatment mediated by the versatile microorganisms. The consortia of cyanobacteria/microalgae and bacteria can be efficient in detoxification of organic and inorganic pollutants, and removal of nutrients from wastewaters, compared to the individual microorganisms. Cyanobacterial/algal photosynthesis provides oxygen, a key electron acceptor to the pollutant-degrading heterotrophic bacteria. In turn, bacteria support photoautotrophic growth of the partners by providing carbon dioxide and other stimulatory means. Competition for resources and cooperation for pollutant abatement between these two guilds of microorganisms will determine the success of consortium engineering while harnessing the biotechnological potential of the partners. Relative to the introduction of gene(s) in a single organism wherein the genes depend on the regulatory- and metabolic network for proper expression, microbial consortium engineering is easier and achievable. The currently available biotechnological tools such as metabolic profiling and functional genomics can aid in the consortium engineering. The present review examines the current status of research on the consortia, and emphasizes the construction of consortia with desired partners to serve a dual mission of pollutant removal and commercial production of microbial metabolites.     

Biofilms of nitrile-hydrolyzing bacteria: Dynamics of growth,resistance to toxic substances,and biotechnological potential

Monospecies and mixed bacterial biofilms of Rhodococcus ruber gt1, Pseudomonas fluorescens C2, Alcaligenes faecalis 2, and Rh. erythropolis 11-2 were obtained during growth in presence of the carriers. The transformation of aliphatic and aromatic nitriles by the biofilms of nitrile-hydrolyzing bacteria, as well as the growth dynamics of Rh. ruber gt1 and P. fluorescens C2 biofilms and their resistance to toxic substrates and products of nitrile hydrolysis, were studied. It was shown that the P. fluorescens C2 biofilm mass and total ATP content reached the maxima after 1 day of growth, whereas Rh. ruber gt1 reached them after 3–4 days of cultivation. The biofilms of Rh. ruber gt1 and P. fluorescens C2 were more resistant to the effects of high concentrations of acrylamide and acrylonitrile and had a greater adaptive capacity than planktonic cells.     

Antimicrobial resistance and in vitro gene transfer in bacteria isolated from the ulcers of EUS-affected fish in India

Aims:  The occurrence of drug resistance and plasmid-mediated transferability was investigated in 15 Aeromonas isolates collected from the ulcers of epizootic ulcerative syndrome (EUS)-affected fishes Katla ( Catla catla ), Mrigel ( Cirrhinus mrigala ) and Punti ( Puntius sp.).
Methods and Results:  Disc diffusion assay showed that all the strains were resistant to ampicillin and sensitive to streptomycin. Of the 15 isolates examined, 93·3% isolates were resistant to erythromycin, sulfadiazine and novobiocin, while 66% were resistant to rifampin and 20% to chloramphenicol. All isolates harboured plasmids with sizes ranging from 64 to 23 kbp with a 23-kbp plasmid in common. Plasmids from 11 Aeromonas strains were transferred to Escherichia coli DH5α recipient strain along with the transfer of ampicillin, erythromycin and chloramphenicol resistance determinants with frequencies ranging from 7·0 × 10⁻⁶ to 1·8 × 10⁻⁵ transconjugants per recipient cell.
Conclusions:  The resistance to ampicillin, erythromycin, sulfadiazine, novobiocin and chloramphenicol is prevalent among the bacteria isolated from EUS-affected fish, and resistant determinants of some of these antibiotics have been transferred to the bacteria of other origin.
Significance and Impact of the Study:  The emergence of antibiotic resistance bacteria and gene transfer in vitr o suggests that antibiotics should be used more cautiously to treat Aeromonas infections in aquaculture.     

Antimicrobial susceptibility of anaerobic bacteria isolated from clinical specimens by using an agar dilution procedure

An agar dilution procedure was used to evaluate the antimicrobial susceptibility of 2,126 anaerobic bacteria isolated from a broad spectrum of properly obtained clinical specimens. Significant numbers ofBacteroides species,Fusobacterium species, anaerobic Gram-positive cocci, and anaerobic nonsporeforming Gram-positive bacilli would not grow on Wilkins-Chalgren agar used in the procedure. The failure to obtain growth from theBacteroides species was of special concern since these organisms generally show a higher degree of resistance to antibiotics than the other anaerobic bacteria, which failed to grow or grew poorly on that medium. When compared to other investigations, the data indicated an emerging resistnace to clindamy a forB. thetaiotaomicron andB. distasonis. IndividualClostridium species showed a wide divergence in susceptibility.     

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.