Antibiotic resistance of soil bacteria

Summary A total of 560 bacterial isolates from four rhizosphere and eight non-rhizosphere soils were examined for resistance to 7 antibiotics. There were marked differences between the overall levels of antibiotic resistance found in the different soils. The rhizosphere populations were not consistently more antibiotic resistant than their corresponding non-rhizosphere soils.     

Antibiotic resistance of raw-milk-associated psychrotrophic bacteria

The present study proposed to evaluate the breadth of antibiotic resistance among psychrotrophic bacteria spoiling raw milks in Finland. A core of 60 isolates, retrieved from farms, trucks, and silos, phenotypically characterized, were analysed with ATB PSE strips, composed of a panel of 17 beta-lactams and non-beta-lactams antibiotics, representatives of five classes. Surprisingly, 60% of the psychrotrophs harboured multiresistant traits. We observed an increase of this feature along the cold chain of raw milk transportation.     

Antibiotic and biocide resistance in bacteria: introduction

Drug resistance in bacteria is increasing and the pace at which new antibiotics are being produced is slowing. It is now almost commonplace to hear about methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), multi-drug resistance in Mycobacterium tuberculosis (MDRTB) strains and multi-drug-resistant (MDR) gram-negative bacteria. So-called new and emerging pathogens add to the gravity of the situation. Reduced susceptibility to biocides is also apparently increasing, but is more likely to be low level in nature and to concentrations well below those used in hospital, domestic an industrial practice. A particular problem, however, is found with bacteria and other micro-organisms present in biofilms, where a variety of factors can contribute to greater insusceptibility compared with cells in planktonic culture. Also of potential concern is the possibility that widespread usage of biocides is responsible for the selection and maintenance of antibiotic-resistant bacteria. The basic mechanisms of action of, and bacterial resistance to, antibiotics are generally well documented, although data continue to accumulate about the nature and importance of efflux systems. In contrast, the modes of action of most biocides are poorly understood and consequently, detailed evaluation of bacterial resistance mechanisms is often disappointing. During this Symposium, the mechanisms of bacterial resistance to antibiotics and biocides are discussed at length. It is hoped that this knowledge will be used to develop newer, more effective drugs and biocides that can be better and perhaps, on occasion, more logically used to combat the increasing problem of bacterial resistance.     

Prevalence of enterohemorrhagic Escherichia coli in raw and treated municipal sewage.

Municipal sewage was screened for DNA encoding Shiga-like Toxin (SLT) II, a key protein involved in the virulence of enterohemorrhagic Escherichia coli. PCR analysis of sewage concentrates showed that DNA encoding SLT II was present in a single sample of untreated sewage and absent in all other samples tested (n = 6). Thermotolerant E. coli cultured from the sewage (n = 1,520) also tested negative for SLT II by colony hybridization.     

Antibiotic resistance of native and faecal bacteria isolated from rivers, reservoirs and sewage treatment facilities in Victoria, south-eastern Australia

The incidence of resistance to ampicillin, chloramphenicol, kanamycin, nalidixic acid, neomycin and streptomycin was significantly greater (P < 0.001) in native heterotrophic bacteria than in Escherichia coli isolated from a range of sites along the Yarra River in south-eastern Australia. There was no significant difference in the incidence of resistance between native and faecal bacteria to tetracycline. Both groups were almost totally resistant to penicillin. Multivariate analyses indicated little clear spatial pattern in the incidence of resistance in native bacteria from upstream vs downstream sites along the Yarra River. In contrast, E. coli isolated from upstream (rural) sites tended to have a lower incidence of resistance than isolates from downstream (urban) sites. These findings have implications for the use of antibiotic resistance as a bacteriological water quality parameter.     

Antibiotic resistance in bacteria and its future for novel antibiotic development

Since the first introduction of the sulfa drugs and penicillin into clinical use, large numbers of antibiotics have been developed and hence contributed to human health. But extensive use of antibiotics has raised a serious public health problem due to multiantibiotic resistant bacterial pathogens that inevitably develop resistance to every new drug launched in the clinic. Consequently, there is a pressing need to develop new antibiotics to keep pace with bacterial resistance. Recent advances in microbial genomics and X-ray crystallography provide opportunities to identify novel antibacterial targets for the development of new classes of antibiotics and to design more potent antimicrobial compounds derived from existing antibiotics respectively. To prevent and control infectious diseases caused by multiantibiotic resistant bacteria, we need to understand more about the molecular aspects of the pathogens' physiology and to pursue ways to prolong the life of precious antibiotics.     

Antibiotic resistance in the bacteria of a remote upland river catchment

The incidence of antibiotic resistance was determined in a total of 183 heterotrophic bacteria of non-faecal origin isolated from the catchment of a remote upland stream. Some 40% were sensitive to each of the eight antibiotics tested while more than 35% were resistant to at least two compounds. Three isolates each carried seven resistance determinants. Unsuccessful attempts were made, using two curing techniques, to demonstrate that resistance was plasmid-borne. The high incidence of antibiotic resistance, particularly amongst pseudomonads, in an environment virtually free from anthropogenic influence is surprising, and has implications for the assessment of the spread of antibiotic resistance amongst populations of environmental bacteria.     

Occurrence of Cryptosporidium spp. oocysts in raw and treated sewage and river water in north-eastern Spain

AIMS: To determine the occurrence and levels of Cryptosporidium parvum oocysts in wastewater and surface waters in north-eastern Spain. METHODS AND RESULTS: Samples from five sewage treatment plants were taken monthly and quarterly during 2003. In addition, water was collected monthly from the River Llobregat (NE Spain) during the period from 2001 to 2003. All samples were analysed by filtration on cellulose acetate filters or through Envirocheck using EPA method 1623, followed by immunomagnetic separation and examination by laser scanning cytometry. All raw sewage, secondary effluent and river water samples tested were positive for Cryptosporidium oocysts. Of the tertiary sewage effluents tested, 71% were positive for Cryptosporidium oocysts. The proportion of viable oocysts varied according to the sample. CONCLUSIONS: Two clear maxima were observed during spring and autumn in raw sewage, showing a seasonal distribution and a correlation with the number of cryptosporidiosis cases and rainfall events. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides the first data on the occurrence of Cryptosporidium oocysts in natural waters in north-eastern Spain.     

Antibiotic resistance of enterococci and related bacteria in an urban wastewater treatment plant

The main objective of this work was to study the ecology of enterococci and related bacteria in raw and treated wastewater from a treatment plant receiving domestic and pretreated industrial effluents in order to assess the influence of treatment on the prevalence of antibiotic resistance phenotypes among this group of bacteria. The predominant species found in the raw wastewater were Entercoccus hirae, Entercoccus faecium and Entercoccus faecalis. Wastewater treatment led to a reduction in E. hirae (alpha<0.1) and an increase in E. faecium (alpha<0.1); the relative proportions of E. faecalis remained the same in the raw and in the treated wastewater. Among the isolates tested, no vancomycin resistance was observed among the enterococci. Entercoccus faecium and E. faecalis showed resistance prevalence values reaching 33%, 40% and 57% for the antibiotics ciprofloxacin, erythromycin and tetracycline, respectively. Antibiotic-resistant strains of enterococci were not eliminated by wastewater treatment. A positive selection of ciprofloxacin-resistant enterococci was indicated by a significant increase in resistance prevalence (alpha<0.02) in treated wastewater compared with the raw wastewater.     

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.

     

Antibiotic resistance patterns of gram-negative bacteria isolated from environmental sources.

A total of 2,445 gram-negative bacteria belonging to fecal coliform, Pseudomonas, Moraxella, Acinetobacter, and Flavobacterium-Cytophaga groups were isolated from the rivers and bay of Tillamook, Oregon, and their resistances to chloramphenicol (25 microgram/ml), streptomycin (10 microgram/ml), ampicillin (10 microgram/ml), tetracycline (25 microgram/ml), chlortetracycline (25 microgram/ml), oxytetracycline (25 microgram/ml), neomycin (50 microgram/ml), nitrofurazone (12.5 microgram/ml), nalidixic acid (25 microgram/ml), kanamycin (25 microgram/ml), and penicillin G (10 IU/ml) were determined. Among fecal coliforms the bay isolates showed greater resistance to antibiotics than those from tributaries or surface runoff. No such well-defined difference was found among other bacterial groups. The antibiotic resistance patterns of gram-negative bacteria from different sources correlated well, perhaps indicating their common origin. The antibiotic resistance patterns of gram-negative bacteria of different general also correlated well, perhaps indicating that bacteria which share a common environment also share a common mode for developing antibiotic resistance.     

Amplicon-based profiling of bacteria in raw and secondary treated wastewater from treatment plants across Australia

In this study, we investigated the use of Illumina high-throughput sequencing of 16S ribosomal RNA (rRNA) amplicons to explore microbial diversity and community structure in raw and secondary treated wastewater (WW) samples from four municipal wastewater treatment plants (WWTPs A–D) across Australia. Sequence reads were analyzed to determine the abundance and diversity of bacterial communities in raw and secondary treated WW samples across the four WWTPs. In addition, sequence reads were also characterized to phenotypic features and to estimate the abundance of potential pathogenic bacterial genera and antibiotic-resistant genes in total bacterial communities. The mean coverage, Shannon diversity index, observed richness (S _obs), and abundance-based coverage estimate (ACE) of richness for raw and secondary treated WW samples did not differ significantly (P > 0.05) among the four WWTPs examined. Generally, raw and secondary treated WW samples were dominated by members of the genera Pseudomonas, Arcobacter, and Bacteroides. Evaluation of source contributions to secondary treated WW, done using SourceTracker, revealed that 8.80–61.4% of the bacterial communities in secondary treated WW samples were attributed to raw WW. Twenty-five bacterial genera were classified as containing potential bacterial pathogens. The abundance of potentially pathogenic genera in raw WW samples was higher than that found in secondary treated WW samples. Among the pathogenic genera identified, Pseudomonas and Arcobacter had the greatest percentage of the sequence reads. The abundances of antibiotic resistance genes were generally low (<0.5%), except for genes encoding ABC transporters, which accounted for approximately 3% of inferred genes. These findings provided a comprehensive profile of bacterial communities, including potential bacterial pathogens and antibiotic-resistant genes, in raw and secondary treated WW samples from four WWTPs across Australia and demonstrated that Illumina high-throughput sequencing can be an alternative approach for monitoring WW quality in order to protect environmental and human health.

     

Antibiotic resistance of hemolytic bacteria, depending on iron content in bacterial cells

The study of sensitivity to antibiotic substances of different chemical structure and the mechanism of the antimicrobial action of hemolytic bacteria Bacillus sp., Corynebacterium sp. and Escherichia coli under the condition of high iron content in bacterial cells was carried out. An increase in antimicrobial resistance of iron-loaded Gram-positive bacteria, in contrast to Gram-negative ones, was shown.     

Antibiotic susceptibility patterns and resistance genes of starter cultures and probiotic bacteria used in food

A survey of starter and probiotic cultures was carried out to determine the current antibiotic resistance situation in microbial food additives in Switzerland. Two hundred isolates from 90 different sources were typed by molecular and other methods to belong to the genera Lactobacillus (74 samples), Staphylococcus (33 samples), Bifidobacterium (6 samples), Pediococcus (5 samples), or were categorized as lactococci or streptococci (82 samples). They were screened for phenotypic resistances to 20 antibiotics by the disk diffusion method. Twenty-seven isolates exhibiting resistances that are not an intrinsic feature of the respective genera were further analyzed by microarray hybridization as a tool to trace back phenotypic resistances to specific genetic determinants. Their presence was finally verified by PCR amplification or Southern hybridization. These studies resulted in the detection of the tetracycline resistance gene tet(K) in 5 Staphylococcus isolates used as meat starter cultures, the tetracycline resistance gene tet(W) in the probiotic cultures Bifidobacterium lactis DSM 10140 and Lactobacillus reuteri SD 2112 (residing on a plasmid), and the lincosamide resistance gene lnu(A) (formerly linA) in L. reuteri SD 2112.     

Anaerobic co-digestion of mechanically biologically treated municipal waste with primary sewage sludge - a feasibility study

This bench scale study investigated the suitability of MBT material for treatment by anaerobic digestion and the impacts of co-digestion of these wastes with sewage sludge. The results suggest that MBT material is amenable to anaerobic digestion with sewage sludge. The main problems for scale-up are related to the physical composition of the MBT material, the accumulation of heavy metals and other inert contaminants and the impact of both of these factors on final sludge quality. Full-scale trials would be required to assess the long-term impacts of MBT waste on anaerobic digestion, if this form of co-digestion were to be pursued. The material contamination that presents a barrier to the direct recycling of MBT material in land-applications is also a major hurdle in commercial co-digestion. Better quality input material would be likely to result in higher methane yields and fewer restrictions on the utilisation of the product in recycling.