Microbiological characterization of aquatic microbiomes targeting taxonomical marker genes and antibiotic resistance genes of opportunistic bacteria

Alexander, Johannes; Bollmann, Anna; Seitz, Wolfram; Schwartz, Thomas

doi:10.1016/j.scitotenv.2015.01.046

Cited by 108 publications

(63 citation statements)

References 49 publications

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“…Absolute quantification of resistance genes in water samples was carried out for four different genes, namely, ampC, vanA, tet(A), and mecA, using the StepOnePlus real-time PCR system (Life Technologies Inc., Burlington, ON, Canada), following a previously described method with slight modifications (25). DNA samples were diluted to a concentration of 0.724 ng/l, and 2.68 l of the diluted DNA (to a total of 1.94 ng of DNA per reaction) was used in a total reaction volume of 8 l which contained 9 M respective primers and 2ϫ SsoFast EvaGreen Supermix (Bio-Rad Canada, Mississauga, ON, Canada).…”

Section: Methodsmentioning

confidence: 99%

“…Although there are other known methods that either calculate the relative quantification of the 16S rRNA gene copy number or absolute quantification of the target gene per volume of the water sample , we used calibration curves derived from different antibiotic resistance reference bacteria for absolute quantification. We prefer this method, which has been used previously (25), because our samples were likely to contain uncharacterized bacteria and also contain bacteria with different genetic backgrounds (e.g., multicopy plasmids), and, therefore, we believe our method of quantifying resistance genes normalizing against the amount of DNA circumvents the possible problems associated with mismatches in the 16S qPCR primers. Multiplex PCR detection of ␤-lactamase and carbapenemase genes.…”

Section: ϫ21mentioning

confidence: 99%

“…2C). The presence of mecA is indicative of methicillin-resistant Staphylococcus aureus (MRSA), which has been shown to be present in various aquatic environments (25,41), although it has also been shown to be present in nonstaphylococcal pathogens (43). mecA was primarily detected in the lake water and bucket samples and in one cistern sample.…”

Section: Fig 2 Quantification Of Ampc (A) Tet(a) (B)mentioning

confidence: 99%

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Detection of Antibiotic Resistance Genes in Source and Drinking Water Samples from a First Nations Community in Canada

Fernando

Tun

Poole

et al. 2016

Appl Environ Microbiol

114

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Access to safe drinking water is now recognized as a human right by the United Nations. In developed countries like Canada, access to clean water is generally not a matter of concern. However, one in every five First Nations reserves is under a drinking water advisory, often due to unacceptable microbiological quality. In this study, we analyzed source and potable water from a First Nations community for the presence of coliform bacteria as well as various antibiotic resistance genes. Samples, including those from drinking water sources, were found to be positive for various antibiotic resistance genes, namely, ampC, tet(A), mecA, ␤-lactamase genes (SHV-type, TEM-type, CTX-M-type, OXA-1, and CMY-2-type), and carbapenemase genes (KPC, IMP, VIM, NDM, GES, and OXA-48 genes). Not surprisingly, substantial numbers of total coliforms, including Escherichia coli, were recovered from these samples, and this result was also confirmed using Illumina sequencing of the 16S rRNA gene. These findings deserve further attention, as the presence of coliforms and antibiotic resistance genes potentially puts the health of the community members at risk. IMPORTANCEIn this study, we highlight the poor microbiological quality of drinking water in a First Nations community in Canada. We examined the coliform load as well as the presence of antibiotic resistance genes in these samples. This study examined the presence of antibiotic-resistant genes in drinking water samples from a First Nations Community in Canada. We believe that our findings are of considerable significance, since the issue of poor water quality in First Nations communities in Canada is often ignored, and our findings will help shed some light on this important issue.A ntibiotic resistance in bacteria has been recognized as one of the greatest threats to human health by the World Health Organization (1). Overuse and misuse of antibiotics contribute to the buildup of selective pressure aiding the proliferation of antibiotic-resistant bacteria (2, 3). While hospital environments are notorious for selecting for antibiotic-resistant bacteria, it is now becoming increasingly evident that overuse and misuse of antibiotics are also creating a selective pressure outside hospital settings. Studies over the last few years have shown the presence of antibiotics and of antibiotic-resistant bacteria in the broader environment, including water supplies and soil samples (4). This is indeed alarming as the high number of antibiotic-resistant bacteria in communities makes the treatment of community-acquired infections increasingly challenging (5, 6).Not surprisingly, water samples from communities that lack access to clean water contain high numbers of bacteria (7-9). While a high bacterial count in the water supply itself poses an increased health risk (10), the presence of antibiotic-resistant bacteria makes this risk even more serious. Lack of access to clean and safe water is a problem that is generally associated with developing countries; however, this is a reality as wel...

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Section: Methodsmentioning

confidence: 99%

Section: ϫ21mentioning

confidence: 99%

Section: Fig 2 Quantification Of Ampc (A) Tet(a) (B)mentioning

confidence: 99%

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Detection of Antibiotic Resistance Genes in Source and Drinking Water Samples from a First Nations Community in Canada

Fernando

Tun

Poole

et al. 2016

Appl Environ Microbiol

114

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“…The definition of drug-resistant bacteria given by ECDC [3] is based on the limit values relating to the clinical strains. That is why it seems necessary to have an alternative approach to the microbial characteristics of environmental strains [4]. However, so far there is no exhaustive data allowing to assess the consequences of the occurence of antibiotic resistance in environmental bacteria.…”

Section: Introductionmentioning

confidence: 99%

Impact of type of wastewater treatment process on the antibiotic resistance of bacterial populations

et al. 2017

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Abstract. Antibiotic resistant bacteria reach the environment directly with faeces, and indirectly with sewage discharged from wastewater treatment plants (WWTPs). The aim of the study was to determine the level of removal of bacteria during wastewater treatment. Samples of untreated and treated sewage were collected from 13 WWTPs with different capacity, modification of treatment and type of inflowing wastewater. Microbiological characterization of the samples included determination of the total number of bacteria resistant (ARB) to β-lactams and tetracyclines as well as the number of Escherichia coli resistant to the same drugs. The counts of ARB and E. coli were determined on TSA and mFc media with/without antibiotic supplementation, respectively. The highest percent of reduction in number of ARB (at least 99.9%) and E. coli (above 99.3%) was obtained for WWTPs with A 2 O system. The lowest percentage reduction of ARB and E. coli was observed for WWTPs operating with SBR system. The lowest number of microorganisms resistant to analyzed antibiotics was observed for ARB and E. coli resistant to cefotaxime and doxycycline. The results indicate a large variation in the removal of antibiotic-resistant bacteria in WWTPs depending on modifications of treatment system and type of inflowing wastewater.

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“…Studies around the world have infrequently and inconsistently detected vanA genes and Enterococcus species isolates with vanA phenotypes in WWTP effluent and surface waters (56)(57)(58)(59). One study in the United States isolated Enterococcus faecium carrying vanA genes on a recreational marine beach in Washington (52), but no other confirmation has been established outside hospital settings.…”

mentioning

confidence: 99%

Vancomycin-Resistant Enterococci and Bacterial Community Structure following a Sewage Spill into an Aquatic Environment

Young

Sun

Badgley

et al. 2016

Appl Environ Microbiol

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Sewage spills can release antibiotic-resistant bacteria into surface waters, contributing to environmental reservoirs and potentially impacting human health. Vancomycin-resistant enterococci (VRE) are nosocomial pathogens that have been detected in environmental habitats, including soil, water, and beach sands, as well as wildlife feces. However, VRE harboring vanA genes that confer high-level resistance have infrequently been found outside clinical settings in the United States. This study found culturable Enterococcus faecium harboring the vanA gene in water and sediment for up to 3 days after a sewage spill, and the quantitative PCR (qPCR) signal for vanA persisted for an additional week. Culturable levels of enterococci in water exceeded recreational water guidelines for 2 weeks following the spill, declining about five orders of magnitude in sediments and two orders of magnitude in the water column over 6 weeks. Analysis of bacterial taxa via 16S rRNA gene sequencing showed changes in community structure through time following the sewage spill in sediment and water. The spread of opportunistic pathogens harboring high-level vancomycin resistance genes beyond hospitals and into the broader community and associated habitats is a potential threat to public health, requiring further studies that examine the persistence, occurrence, and survival of VRE in different environmental matrices. IMPORTANCEVancomycin-resistant enterococci (VRE) are harmful bacteria that are resistant to the powerful antibiotic vancomycin, which is used as a last resort against many infections. This study followed the release of VRE in a major sewage spill and their persistence over time. Such events can act as a means of spreading vancomycin-resistant bacteria in the environment, which can eventually impact human health.A ntibiotic-resistant bacteria (ARB) are a growing public health threat and an economic burden globally. The Centers for Disease Control and Prevention (CDC) in the United States has placed a high priority on addressing antibiotic resistance because of rising rates of ARB infection and associated disease burden and health care costs (1, 2). Most infections caused by ARB are nosocomial transmissions (i.e., originating in a hospital), but the role of environmental reservoirs in spreading ARB outside clinical settings is poorly understood. Studies have emphasized the role of environmental reservoirs in the spread of antibiotic resistance for decades, but more field and laboratory studies are necessary to address the specific mechanisms and conditions under which ARB survive and antibiotic resistance genes (ARGs) persist or can be transferred (3-5). Wastewater treatment plants (WWTPs) are sources of ARB, ARGs, and antimicrobial compounds through both treated effluent and the unplanned release of raw sewage to surface waters (6-9). ARB, ARGs, and antibiotics can be released into aquatic environments through human and agricultural waste, establishing routes of human exposure and threats to ecosystem health.Vancomycin i...

show abstract

Microbiological characterization of aquatic microbiomes targeting taxonomical marker genes and antibiotic resistance genes of opportunistic bacteria

Cited by 108 publications

References 49 publications

Detection of Antibiotic Resistance Genes in Source and Drinking Water Samples from a First Nations Community in Canada

Detection of Antibiotic Resistance Genes in Source and Drinking Water Samples from a First Nations Community in Canada

Impact of type of wastewater treatment process on the antibiotic resistance of bacterial populations

Vancomycin-Resistant Enterococci and Bacterial Community Structure following a Sewage Spill into an Aquatic Environment

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