Extended Spectrum Beta-Lactamase (ESBL) Producing Enterobacteria in Aquatic Environmental Sources of Bangladesh (original) (raw)

Extended spectrum beta-lactamase (ESBL) producing escherichia coli in poultry and water in Lusaka district

2015

Untreated wastewater is a risk factor for the spread of antibiotic resistance in the environment. However, little is known about the contribution of untreated wastewater to the burden of antibiotic resistance in the Nigerian environment. In this study, a total of 143 ceftazidime-/cefpodoxime-resistant bacteria isolated from untreated wastewater and untreated wastewater-contaminated surface and groundwater in Nigeria were screened for extended-spectrum β-lactamase (ESBL) genes, integrons and integron gene cassettes by PCR. The genetic environment of bla CTX-M-15 was mapped by PCR and potentially conjugative plasmids were detected among the isolates by degenerate primer MOB typing (DPMT). ESBL production was confirmed in 114 (79.7%) isolates and ESBL genes (bla SHV , bla CTX-M-15 and bla TEM) were detected in 85 (74.6%) ESBL-producing isolates. bla CTX-M-15 was associated with ISEcp1 and with orf477 in 12 isolates and with ISEcp1, IS26 and orf477 in six others. To the best of our knowledge, this is the first report of bla CTX-M-15 in hand-dug wells and borehole serving as sources of drinking water and a first report of the genetic environment of bla CTX-M-15 in environmental bacteria from Nigeria. The results of this study confirm untreated wastewater as an important medium for the spread of ESBL-producing bacteria within the Nigerian environment. Hence, the widespread practice of discharging untreated wastewater into the aquatic ecosystem in Nigeria is a serious risk to public health. Keywords Antibiotic resistance. β-Lactamases. bla SHV. bla TEM. bla CTX-M-15. Conjugative relaxase. Untreated wastewater

A survey of extended-spectrum beta-lactamase-producing Enterobacteriaceae in urban wetlands in southwestern Nigeria as a step towards generating prevalence maps of antimicrobial resistance

PLOS ONE

In many countries, emission of insufficiently treated wastewater into water bodies appears to be an important factor in spreading clinically relevant antimicrobial resistant bacteria. In this study, we looked for the presence of Enterobacteriaceae strains with resistance to 3 rd generation cephalosporin antibiotics in four urban wetlands in southwestern Nigeria by isolation, whole genome sequencing and qPCR enumeration of marker genes. Genome analysis of multi-drug resistant and potentially pathogenic Escherichia coli isolates (members of the widely distributed ST10 complex) revealed the presence of the extended spectrum beta-lactamase gene bla CTX-M-15 on self-transmissible IncF plasmids. The gene was also present together with a bla TEM-1B gene on self-transmissible IncH plasmids in multi-drug resistant Enterobacter cloacae isolates. A Citrobacter freundii isolate carried bla TEM-1B on an IncRtype plasmid without discernable conjugation apparatus. All strains were isolated from a wetland for which previous qPCR enumeration of marker genes, in particular the ratio of intI1 to 16S rRNA gene copy numbers, had indicated a strong anthropogenic impact. Consistent with the isolation origin, qPCR analysis in this study showed that the bla CTX-M gene was present at an abundance of 1x10-4 relative to bacterial 16S rRNA gene copy numbers. The results indicate that contamination of these urban aquatic ecosystems with clinically relevant antibiotic resistant bacteria is substantial in some areas. Measures should therefore be put in place to mitigate the propagation of clinically relevant antimicrobial resistance within the Nigerian aquatic ecosystems.

Resistotyping and extended-spectrum beta-lactamase genes among Escherichia coli from wastewater treatment plants and recipient surface water for reuse in South Africa

New Microbes and New Infections, 2020

The spread of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli has increased in different environments. This study aimed to evaluate the patterns of antibiotic resistance and ESBL genes among E. coli isolates collected from wastewater and recipient surface water in South Africa. Fifteen samples containing nine wastewater and six river water samples were collected from a local wastewater treatment plant. The E. coli isolates were detected using standard microbiology methods. Antibiotic susceptibility testing was performed using disc diffusion agar. The occurrence of bla CTX-M , bla SHV and bla TEM ESBL genes was investigated by PCR. Exactly 140 isolates were selected from the primary enumeration plates with a log 10 CFU/mL count that ranged from 4.1 to 4.2 (influent), 4.2 to 4.5 (biofilter) and 2.5 to 3.3 (effluent). The wastewater effluent showed an impact on the receiving water environment, as the treatment efficiency was 92% and the downstream log 10 CFU/mL count (range, 3.6-3.8 log 10 CFU/mL) was higher than the upstream count (range, 3.3-3.6 log 10 CFU/mL). Antibiotic testing results showed that 40% to 100% of E. coli isolates were resistant to ampicillin, penicillin, tetracycline and cefotaxime but susceptible to imipenem, meropenem and ciprofloxacin. A total of 40 studied isolates (28.6%) had both the bla TEM and bla CTX-M genes, while no bla SHV was detected. The wastewater treatment plants contributed multidrug-resistant ESBL-producing E. coli isolates that can be potential environmental health risks. Regular monitoring policies are recommended to prevent the spread of antibiotic resistance in the region.

Characterization of Extended Spectrum Beta-Lactamases (ESBL) Producing Escherichia coli Isolates from Surface Water Adjacent to Pharmaceutical Industries in Bangladesh: Antimicrobial Resistance and Virulence Pattern

Microbiology and Biotechnology Letters

The pharmaceutical industry in Bangladesh produces a diverse range of antibiotics for human and animal use, however, waste disposal management is inadequate. This results in substantial quantities of antibiotics being discharged into water bodies, which provide suitable environment for the growth of antibiotic-resistant bacteria, capable of spreading resistance genes. This study intended for exploring the bacterial antibiotic resistance profile in adjoining aquatic environmental sources of pharmaceutical manufacturing facilities in Bangladesh. Seven surface water samples were collected from the vicinity of two pharmaceutical industries located in the Savar area and 51 Escherichia coli isolates were identified using both phenotypic and genotypic methods. Antibiotic susceptibility tests revealed the highest percentage of resistance against ampicillin, azithromycin, and nalidixic acid (100%) and the lowest resistance against meropenem (1.96%) out of sixteen different antibiotics tested. 100% of the study E. coli isolates were observed with Multidrug resistance phenotypes, with the Multiple Antibiotic Resistance (MAR) value ranging from 0.6-1.0. Furthermore, 69% of the isolates were Extended Spectrum Beta-Lactamases (ESBL) positive as per the Double Disk Diffusion Synergy Test (DDST). ESBL resistance genes bla TEM , bla CTX-M-13 , bla CTX-M-15 , and bla SHV were detected in 70.6% (n = 36), 60.8% (n = 32), 54.9% (n = 28), and 1.96% (n = 1) of the isolates, respectively, by Polymerase Chain Reaction (PCR). Additionally, 15.68% (n = 8) of the isolates were positive for E. coli specific virulence genes in PCR. These findings suggest that pharmaceutical wastewater, if not properly treated, could be a formidable source of antibiotic resistance spread in the surrounding aquatic environment. Therefore, continued surveillance for drug resistance among bacterial populations around drug manufacturing facilities in Bangladesh is necessary, along with proper waste disposal management.

Genetic diversity and prevalence of extended spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae in aquatic environment receiving untreated hospital effluents

Research Square (Research Square), 2022

The spread of extended spectrum beta-lactamase producing bacteria in the environment has been recognized as a challenge to public health. Currently one of the most important mechanisms of resistance in Enterobacteriaceae is the synthesis of enzymes that contribute to modern expanded spectrum cephalosporin resistance, mainly extended spectrum beta-lactamases. The aim of the present study was to assess the occurrence of ESBL producing Escherichia coli and Klebsiella pneumoniae from selected water bodies receiving hospital e uents in Kerala, India. Nearly 69.8% of Enterobacteriaceae isolates were multi-drug resistant by Kirby-Bauer disc diffusion method. Double disc synergy test was used to detect the ESBL production and the genes responsible for imparting resistance was detected by PCR. Conjugation experiments con rmed the mechanism of plasmid-mediated transfer of resistance. The prevalence of ESBL production in E. coli and K. pneumoniae was 37.4% and 11.2% respectively. Among the ESBL encoding genes, bla CTX−M was the most prevalent group followed by bla TEM , bla OXA , bla CMY and bla SHV. The results suggest that healthcare settings are one of the key contributors in the spread of ESBLproducing bacteria, not only through cross-transmission and ingestion of antibiotics, but also through the discharge of waste without a proper treatment, leading to harmful effects on the aquatic environment. The high prevalence of ESBL-producing Enterobacteriaceae with resistance genes in public water bodies even post-treatment poses a serious threat. .

Antibiotic resistance pattern and molecular detection of ESBL-associated genes in E. coli from surface and wastewater of Islamabad capital territory, Pakistan

Journal of Water and Health

The goal of this study was to determine how surface and wastewater contribute to the contamination of the environment with an extended-spectrum β-lactamase-producing Escherichia coli (ESBL E. coli). Water samples (n = 32) were collected from eight different locations of Islamabad and processed for microbiological and molecular analyses of E. coli and ESBL E. coli. Antimicrobial susceptibility testing was carried out to determine the resistance pattern of the isolates. A total of 21 water samples were contaminated with E. coli and 15 isolates were identified as ESBL producers harboring blaTEM (40%) and blaCTX-M (33.33%) genes. Interestingly, all the ESBL E. coli isolates showed the least resistance against second-generation Cephalosporins compared to other generations. Moreover, the study showed that the aquatic environment is harboring multidrug-resistant E. coli; therefore, it may act as a source of transmission to humans. The recovery of ESBL E. coli isolates resistant to higher g...

Distribution Profile of Extended Spectrum Beta Lactamase (ESBL) producing Escherichia coli Isolates from Asa River (Nigeria)

Journal of Applied Sciences and Environmental Management, 2017

Escherichia coli are known pathogenic organism that has caused diseases which has led to severe morbidity and increased death rate. The occurrence of extended spectrum beta Lactamase (bla) producing Escherichia coli has been on the rise. Water samples were investigated as a potential reservoir for the Extended Spectrum Beta-Lactamase (ESBL)-producing E. coli using phenotypic (culture-based) and molecular methods. Double disc synergy test was determined between a disc of amoxicillin-clavulanate (20µg/10µg) (augmentin) and a 30-µg disc of each thirdgeneration cephalosporin antibiotic placed at a distance of 20 mm from centre to centre on a Mueller-Hinton Agar plate streaked with the isolate. An isolate was considered to be ESBL negative if there was no enhancement between any of the cephalosporin and the clavulanate-containing discs and were then subjected to specific Polymerase Chain Reaction (PCR). Eighty-four environmental E. coli was isolated. 58(69.04%) showed positivity for ESBL production. E. coli isolates positive for ESBL-production selected and subjected to plasmid curing were all plasmid mediated. 16 isolates subjected to PCR to identify the presence of blaSHV (Sulphydryl Variable), blaTEM (Temoneira) and blaCTX-M (Cefotaximase) genes revealed that 11(68.7%) of these had at least one ESBL gene (either blaCTX-M or blaTEM, or both), 5(31.3%) isolates do not have any of the three ESBL genes, and blaSHV was not detected in any of the isolates. The results of this study indicate the widespread prevalence of ESBLs in E. coli. Therefore, beta-lactam antibiotics and beta-lactamase inhibitors should be prescribed based on an antibacterial susceptibility test. © JASEM

Characterization of Ciprofloxacin resistant Extended Spectrum <i>β</i>-Lactamase (ESBL) producing <i>Escherichia </i>spp. from clinical waste water in Bangladesh

Advances in Bioscience and Biotechnology, 2013

Clinical Waste Water (CWW) possesses the risks of spreading antibiotic resistant bacteria in the environment. In Bangladesh, liquid discharge is directly released into the municipal sewage system and pollutes the surroundings water bodies/rivers. Liquid samples were collected from the three CWW discharge sites connected to DMCH (Dhaka Medical College Hospital) and from one control group non-connected to DMCH. About 32, 30 and 26 times higher ciprofloxacin, cefixime and multi-drug resistant bacterial count were found in CWW over control samples. Among the isolates, two Escherichia spp. isolates, denoted 26N and 28N, were found to be resistant against fluoroquinolone (MBC of Ciprofloxacin > 1024 μg/ml), cephalosporin, glycopeptide, monobactam, penicillin, tetracycline, rifampicin, macrolides, sulfonamide and nitrofuran classes of drugs and were also ESBL positive through phenotypic assay. Plasmid curing experiment detected possible plasmid mediated resistance of fluoroquinolone, cephalosporin, tetracycline, lincosamide and nitrofuran classes of antibiotics. Phylogenetically, isolate 26N and 28N were characterized as Escherichia coli and Escherichia fergusonii. These MDR and ESBL positive bacteria are potent to disseminate resistant determinants in the surrounding environments.

Extended spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae isolated from dogs

2017

Untreated wastewater is a risk factor for the spread of antibiotic resistance in the environment. However, little is known about the contribution of untreated wastewater to the burden of antibiotic resistance in the Nigerian environment. In this study, a total of 143 ceftazidime-/cefpodoxime-resistant bacteria isolated from untreated wastewater and untreated wastewater-contaminated surface and groundwater in Nigeria were screened for extended-spectrum β-lactamase (ESBL) genes, integrons and integron gene cassettes by PCR. The genetic environment of bla CTX-M-15 was mapped by PCR and potentially conjugative plasmids were detected among the isolates by degenerate primer MOB typing (DPMT). ESBL production was confirmed in 114 (79.7%) isolates and ESBL genes (bla SHV , bla CTX-M-15 and bla TEM) were detected in 85 (74.6%) ESBL-producing isolates. bla CTX-M-15 was associated with ISEcp1 and with orf477 in 12 isolates and with ISEcp1, IS26 and orf477 in six others. To the best of our knowledge, this is the first report of bla CTX-M-15 in hand-dug wells and borehole serving as sources of drinking water and a first report of the genetic environment of bla CTX-M-15 in environmental bacteria from Nigeria. The results of this study confirm untreated wastewater as an important medium for the spread of ESBL-producing bacteria within the Nigerian environment. Hence, the widespread practice of discharging untreated wastewater into the aquatic ecosystem in Nigeria is a serious risk to public health. Keywords Antibiotic resistance. β-Lactamases. bla SHV. bla TEM. bla CTX-M-15. Conjugative relaxase. Untreated wastewater

Analysis of Wastewater Reveals the Spread of Diverse Extended-Spectrum β-Lactamase-Producing E. coli Strains in uMgungundlovu District, South Africa

Antibiotics

Wastewater treatment plants (WWTPs) are major reservoirs of antibiotic-resistant bacteria (ARB), favouring antibiotic resistance genes (ARGs) interchange among bacteria and they can provide valuable information on ARB circulating in a community. This study characterised extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli from the influent and effluent of four WWTPs in uMgungundlovu District, KwaZulu-Natal, South Africa. E. coli was enumerated using the membrane filtration method and confirmed using the API 20E test and real-time polymerase chain reaction. ESBL-producers were phenotypically identified by their susceptibility to the third-generation cephalosporins using the disc diffusion and the double-disc synergy methods against cefotaxime (30 µg) with and without 10 µg clavulanic acid. Genotypic verification was by PCR of the TEM, SHV, and CTX-M genes. The clonality of isolates was assessed by ERIC-PCR. The highest E. coli count ranged between 1.1 × 105 (influent) a...