Prevalence of antibiotic-resistant fecal bacteria in a river impacted by both an antibiotic production plant and urban treated discharges. - Related Documents




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526501.0000Prevalence of antibiotic-resistant fecal bacteria in a river impacted by both an antibiotic production plant and urban treated discharges. In this study, the abundance and spatial dynamics of antibiotic-resistant fecal bacteria (Escherichia coli, total coliforms and Enterococcus spp.) were determined in water and sediment samples from a river impacted by both antibiotic production plant (APP) and urban wastewater treatment plant (WWTP) discharges. Agar dilution and disk diffusion methods were also used for antimicrobial susceptibility testing. Two antimicrobial agents, cephalexin (25 μg/ml) and amoxicillin (50 μg/ml), were evaluated using the agar dilution method for E. coli, total coliforms (TC) and Enterococcus spp., whereas the degree of sensitivity or resistance of E. coli isolates to penicillin (10 U), ampicillin (10 μg), doxycycline (30 μg), tetracycline (30 μg), erythromycin (15 μg), azithromycin (15 μg) and streptomycin (10 μg) was performed using the disk diffusion method. Real-time PCR assays were used to determine the prevalence of three antibiotic-resistance genes (ARGs). The agar dilution method showed that most E. coli isolates and TC were resistant to amoxicillin, especially after receiving the APP discharges. Antibiotic resistances to amoxicillin and cephalexin were higher after the APP discharge point than after the WWTP effluent. The disk diffusion method revealed that 100% of bacterial isolates were resistant to penicillin and erythromycin. Multidrug-resistant bacteria were detected and showed a higher proportion at the WWTP discharge point than those in the APP. Highly multidrug-resistant bacteria (resistance to more than 4 antibiotics) were also detected, reaching mean values of 41.6% in water samples and 50.1% in sediments. The relative abundance of the blaTEM, blaCTX-M and blaSHV genes was higher in samples from the treatment plants than in those collected upstream from the discharges, especially for water samples collected at the APP discharge point. These results clearly demonstrate that both the APP and the WWTP contribute to the emergence and spread of antibiotic resistance in the environment.201424836130
527110.9999Occurrence of Antibiotic-Resistant Bacteria and Genes in Two Drinking Water Treatment and Distribution Systems in the North-West Province of South Africa. With the increasing spread of antimicrobial resistance, there is growing attention to the contribution made by drinking water systems. The potential health impact of two drinking water treatment and distribution systems (A and B) in the North-West Province of South Africa was determined by investigating the water quality and occurrence of antimicrobial-resistant heterotrophic bacteria and genes in the raw and treated water over four seasons. Most of the physicochemical parameters except for electrical conductivity were within permissible limits. Coliform bacteria reduced from raw to potable water except for counts higher than the threshold recorded in Summer and Winter. A total of 203 heterotrophic bacterial isolates were recovered on chromogenic R2A medium and subjected to susceptibility testing to twelve antibiotics. Most of the isolates were resistant to β-lactam antibiotics and Trimethoprim, whereas they were susceptible to Ciprofloxacin, Erythromycin, and Neomycin. The proportions of Cephalothin and Kanamycin-resistant isolates were significantly higher (p < 0.05) after treatment for site A, compared to significantly lower β-lactam, Oxytetracycline, and Trimethoprim-resistant isolates for B. Over 50% of isolates were of high risk, indicating their origin from high antibiotic-use sources. Seventy-one (35%) isolates were multidrug-resistant, out of which the majority (53.5%, n = 38) possessed the strA gene, followed by strB 21 (29.6%), dfrB 13 (18.3%), aadA 11 (15.5%), bla(CTX-M) 5 (7.0%), and tetA 3 (4.2%). The 16S rRNA gene sequences of the isolates revealed strains belonging to eight bacterial families, some of which are clinically important.202033126462
527020.9999Environmental antibiotic stress and high-risk resistance genes in bacterial communities of the Gomti and Ganga Rivers, India. BACKGROUND: The river ecosystems provide habitats and source of water for a number of species including humans. The uncontrolled accumulation of pollutants in the aquatic environment enhances the development of antibiotic-resistant bacteria and genes. METHODS: Water samples were collected seasonally from different sites of Gomti and Ganga River. Bacteria were isolated by plating on nutrient agar supplemented with individual antibiotics (100 µg/ml) to select the resistant strains. These isolates were subsequently tested for cross-resistance to other antibiotics using the disc diffusion method. PCR was performed to detect selected ARGs. RESULTS: The enumeration of microbial population of Gomti River, the tetracycline-resistant bacteria comprised 38% of the bacterial population during spring and chloramphenicol resistance during autumn was a mere 11.9%. Nevertheless, erythromycin resistance was widespread amongst Ganga river bacteria during winter by 28%, while ciprofloxacin resistance was seen in autumn with only 15.8%. Bacterial population led to decline due to antibiotic-induced stress. The tetracycline-resistant bacteria were completely resistant to ampicillin and 66.6% were resistant to erythromycin. In Ganga river water, 53.5% of ampicillin-resistant isolates were resistant to erythromycin and sulphadiazine 93.3% were resistant to nalidixic acid. In the Gomti River water, the most common resistance gene among tetracycline resistant isolates was tetM (83.3%), followed by ampC (83.3%) in ampicillin-resistant isolates. In the Ganga River, 66.6% of bacterial isolates were found to have ampC and ermB genes. The sul1 gene was absent in all the bacterial isolates in both water samples. CONCLUSION: These findings indicate that both rivers act as reservoirs for multidrug-resistant bacteria harbouring complex resistance gene profiles.202540928717
525630.9999Characterization of antibiotic resistance genes and bacteria in a municipal water resource recovery facility. Municipal water resource recovery facilities (WRRFs) are important sources of antibiotic-resistant bacteria and genes (ARB and ARGs). In this study, antibiotic-resistant total heterotrophic bacteria (THB(R) ) counts (CFU/ml) cultivated from influent, effluent of activated sludge process, and outflow of disinfection unit of an urban WRRF were investigated for the presence of 16, 32, 64, and 128 μg/ml of nine antibiotics. The isolates of Pseudomonas spp., Acinetobacter spp., and Escherichia coli obtained from effluent of activated sludge process were subjected for molecular identification by detecting the 16S rRNA gene sequences. Additionally, using the polymerase chain reaction method (PCR), the isolates were investigated for the presence of bla(SHV) , bla(TEM) , bla(CTX-M) , bla(VIM) , sul1, and qnrS genes. According to the results, the abundance of THB(R) counts was not significantly reduced by the biological treatment except for cefixime and sulfamethoxazole; it also increased for some antibiotics after disinfection unit. The average removal efficiency of THB(R) resistant to ciprofloxacin, sulfamethoxazole, and ceftazidime were 7.9 ± 1.7%, 41.8 ± 2.1%, and 14.4 ± 6.2%, respectively. Also, all the tested isolates were resistant to at least four antibiotics. For all antibiotics, the resistance ratio (THB(R) /THB) significantly increased in the effluent and after chlorination unit. Among 12 resistant isolates, bla(TEM) and sul1 genes were the most frequently detected ones involved in 92% and 83% of the isolates, respectively. Both bla(TEM) and sul1 genes were found in 100% of E. coli, and 83% and 67% of Pseudomonas spp. isolates, respectively. Further efforts are necessary to limit the transmission of ARB and ARGs from WRRFs into the environment and prevent human health threats. PRACTITIONER POINTS: The ratio of resistance significantly increased after biological treatment. Up to 40% of heterotrophic bacteria in the effluent was antibiotic resistant. bla(TEM) and sul1 genes were more prevalent (92%) in all isolates of bacteria. Both bla(TEM) and sul1 genes were found in 100% of E. coli isolates. Pseudomonas spp. holds bla(TEM) and sul1 genes in 83% and 67% of isolates, respectively.202235765862
277140.9998Identification, antibiotic resistance, and virulence profiling of Aeromonas and Pseudomonas species from wastewater and surface water. Aquatic environments are hotspots for the spread of antibiotic-resistant bacteria and genes due to pollution caused mainly by anthropogenic activities. The aim of this study was to evaluate the impact of wastewater effluents, informal settlements, hospital, and veterinary clinic discharges on the occurrence, antibiotic resistance profile and virulence signatures of Aeromonas spp. and Pseudomonas spp. isolated from surface water and wastewater. High counts of Aeromonas spp. (2.5 (± 0.8) - 3.3 (± 0.4) log(10) CFU mL(-1)) and Pseudomonas spp. (0.6 (± 1.0) - 1.8 (± 1.0) log(10) CFU mL(-1)) were obtained. Polymerase chain reaction (PCR) and MALDI-TOF characterization identified four species of Aeromonas and five of Pseudomonas. The isolates displayed resistance to 3 or more antibiotics (71% of Aeromonas and 94% of Pseudomonas). Aeromonas spp. showed significant association with the antibiotic meropenem (χ(2) = 3.993, P < 0.05). The virulence gene aer in Aeromonas was found to be positively associated with the antibiotic resistance gene blaOXA (χ(2) = 6.657, P < 0.05) and the antibiotic ceftazidime (χ(2) = 7.537, P < 0.05). Aeromonas recovered from both wastewater and surface water displayed high resistance to ampicillin and had higher multiple antibiotic resistance (MAR) indices close to the hospital. Pseudomonas isolates on the other hand exhibited low resistance to carbapenems but very high resistance to the third-generation cephalosporins and cefixime. The results showed that some of the Pseudomonas spp. and Aeromonas spp. isolates were extended-spectrum β-lactamase producing bacteria. In conclusion, the strong association between virulence genes and antibiotic resistance in the isolates shows the potential health risk to communities through direct and indirect exposure to the water.202133893564
285550.9998Antibiotic resistance, plasmid-mediated quinolone resistance (PMQR) genes and ampC gene in two typical municipal wastewater treatment plants. Antibiotic resistant bacteria and plasmid-mediated quinolone resistance genes and ampC gene were investigated for Escherichia coli isolates from two typical municipal wastewater treatment plants in both dry and wet seasons by using the antibiotic susceptibility test and PCR assay, respectively. The results showed that 98.4% of the isolates (1056) were found resistant to antibiotic(s) tested and 90.6% showed multiple resistances to at least three antibiotics. Tetracycline was found to have the highest resistance frequency (70.8%), followed by ampicillin (65.1%), whereas ceftazidime had the lowest resistance frequency of 9.0%. Moreover, 39.2% of the E. coli isolates were carrying plasmids. intI1 had the highest detection rate in the plasmids (38.1%), followed by qnrS, ampC, qnrB, intI2 and aac(6')-Ib-cr. The disinfection process (UV and chlorination) could significantly reduce the number of bacteria, but percentage of the resistant bacteria, resistance frequency for each antibiotic, MAR index and detection rate of the plasmid-mediated resistance genes were all found increasing in the effluents of biological units. The results of this study showed that a more frequent horizontal gene transfer occurred in the biological units. Wastewater treatment plants were an important medium for the recombination and dissemination of antibiotic resistance genes in the environment.201424441525
135960.9998Assessment of Bacterial Contamination and Antimicrobial Resistance of Escherichia coli Isolates from Slovak Dairy Farms. The conditions in livestock housing are suitable for the survival of airborne microorganisms, mainly due to high temperatures, humidity, and the presence of organic material. The total count of airborne bacteria concentrations in cattle farms ranged from 3.01 log(10) CFU/mL to 6.90 log(10) CFU/mL; for coliform bacteria, they were from 2.18 log(10) CFU/mL to 3.34 log(10) CFU/mL; and for molds, they ranged from 3.00 log(10) CFU/mL to 4.57 log(10) CFU/mL. Bacteria resistant to antimicrobial substances and resistance genes can be spread on animal farms. Antimicrobial resistance in ubiquitous Escherichia coli isolated from cattle feces was investigated. Minimum inhibitory concentration (MIC) testing was utilized to identify phenotypic resistance profiles, and the PCR method was employed to detect the presence of resistant genes. A higher percentage of resistance was found to amikacin (65%), tetracycline (61%), streptomycin (56%), ampicillin (55%), and nalidixic acid (45%). Multidrug resistance was determined in up to 64.3% of the isolates studied. The most widespread resistance genes were bla(TEM) (85.7%), sul2 (66.7%), tetB (52.38%), and sul1 (47.6%). We found that 4.8% of the E. coli isolates had the bla(CMY) gene. We found that, despite phenotypic resistance, E. coli isolates do not necessarily carry genes conferring resistance to that particular antimicrobial agent.202439518818
273770.9998Meropenem-resistant bacteria in hospital effluents in Seoul, Korea. This study aimed to understand the prevalence, diversity, antibiotic resistance, β-lactamase gene types, and possibility of environmental survival of meropenem-resistant bacteria present in hospital effluents in Seoul, Korea. Water sampling was performed at five general hospitals in Seoul, Korea, in January 2017. Water samples were plated in triplicate on tryptic soy agar plates with 16 mg/L meropenem. Meropenem-resistant bacteria were selected and subjected to 16S rRNA analysis for species determination and PCR for identification of β-lactamase gene types. Resistant bacteria were cultured in sterilized surface water. Meropenem-resistant bacteria exhibited resistance to more than 12 antibiotics and possessed several β-lactamase genes, such as those encoding OXT-M, NDM-1, AmpC, and OXA. They were able to multiply and survive in sterilized surface water for up to 60 days. Multidrug-resistant bacteria represent an environmental health risk, as they can survive in the environment for an extended period of time. Therefore, these bacteria should be monitored before discharge.201830361772
286980.9998Antibiotic resistance and antibiotic-resistance genes of Pseudomonas spp. and Escherichia coli isolated from untreated hospital wastewater. Hospitals are considered an important factor in the spread of antibiotic-resistant bacteria (ARBs) and antibiotic-resistance genes (ARGs). The purpose of this research was to characterize the microbial populations in hospital wastewater and investigated the prevalence of β-lactamase, SulІ and QnrS resistance genes. In the first step, culture method was used to isolate Pseudomonas aeruginosa and Escherichia coli. In the next step, accurate identification of isolated bacteria was carried out using the polymerase chain reaction (PCR) method, then the resistance of the bacteria at different concentrations of antibiotics (8-128 μg/mL) was examined. Finally the ARGs were detected using the PCR method. The averages of heterotrophic plate count (HPC) and ARB concentration in wastewater samples were 1.8 × 10(8) and 4.3 × 10(6) CFU/100 mL, respectively. The highest resistance rates were found for sulfamethoxazole and the highest resistance rates in the β-lactamase group were for ceftazidime, while highest sensitivity was for gentamicin and there was no isolate that was sensitive to the studied antibiotics. SulІ and QnrS were the highest and lowest abundance of all ARGs in samples respectively and blaSHV was the highest β-lactam resistance gene. Our results indicated an increase in the resistance of identified bacteria to several antibiotics. So it can be concluded that numerous antibiotic-resistant pathogens and vast numbers of ARGs exist in the human body so that their release from hospitals without effective treatment can cause many dangers to the environment and human health.202134280162
526690.9998Distribution analysis of tetracycline resistance genes in Escherichia coli isolated from floor surface and effluent of pig slaughterhouses in Banten Province, Indonesia. BACKGROUND AND AIM: Slaughterhouses and their effluents could serve as a "hotspot" for the occurrence and distribution of antibiotic-resistant bacteria in the environment. This study aimed to understand the distribution of tetracycline resistance genes in Escherichia coli isolated from the floor surface and effluent samples of pig slaughterhouses in Banten Province, Indonesia. MATERIALS AND METHODS: Ten samples, each from floor surface swabs and effluents, were collected from 10 pig slaughterhouses in Banten Province. Escherichia coli strains were isolated and identified by referring to the protocol of the Global Tricycle Surveillance extended-spectrum beta-lactamase E. coli from the WHO (2021). Quantitative real-time polymerase chain reaction (qPCR) was used to detect the tet genes. RESULTS: The tetA, tetB, tetC, tetM, tetO, and tetX genes were distributed in the isolates from the floor surface samples, and the tetA, tetC, tetE, tetM, tetO, and tetX genes were distributed in the isolates from the effluent samples. The tetO gene (60%) was the most dominant gene in the isolates from floor surface samples, while the tetA gene was the dominant one in the isolates from the effluent samples (50%). The tetA + tetO gene combination was the dominant pattern (15%) in the E. coli isolates. CONCLUSION: The high prevalence and diversity of the tet genes in floor surface and effluent samples from pig slaughterhouses in Banten Province indicated that the transmission of the tet genes had occurred from pigs to the environment; thus, this situation should be considered a serious threat to public health.202337041843
2736100.9998Characterization of Bacterial Communities and Their Antibiotic Resistance Profiles in Wastewaters Obtained from Pharmaceutical Facilities in Lagos and Ogun States, Nigeria. In Nigeria, pharmaceutical wastewaters are routinely disseminated in river waters; this could be associated with public health risk to humans and animals. In this study, we characterized antibiotic resistant bacteria (ARB) and their antibiotic resistance profile as well as screening for sul1 and sul2 genes in pharmaceutical wastewater effluents. Bacterial composition of the wastewater sources was isolated on non-selective media and characterized by the polymerase chain reaction (PCR) amplification of the 16S rRNA genes, with subsequent grouping using restriction fragment length polymorphism (RFLP) and sequencing. The antibiotics sensitivity profiles were investigated using the standard disk diffusion plate method and the minimum inhibitory concentrations (MICs) of selected antibiotics on the bacterial isolates. A total of 254 bacterial strains were isolated, and majority of the isolates were identified as Acinetobacter sp., Klebsiella pneumonia, Proteus mirabilis, Enterobacter sp. and Bacillus sp. A total of 218 (85.8%) of the bacterial isolates were multidrug resistant. High MICs values were observed for all antibiotics used in the study. The result showed that 31.7%, 21.7% and 43.3% of the bacterial isolates harbored sul1, sul2, and Intl1 genes, respectively. Pharmaceuticals wastewaters are potential reservoirs of ARBs which may harbor resistance genes with possible risk to public health.201829966226
2858110.9998Antibiotic resistance in fecal sludge and soil in Ho Chi Minh City, Vietnam. This study investigated the prevalence of antibiotic-resistant bacteria and genes in fecal sludge and soil in Ho Chi Minh City, Vietnam, and identified the factors contributing to the survival of antibiotic-resistant bacteria in soil. Sludge and soil samples (n = 24 and 55, respectively) were collected from residential septic systems and environmental reservoirs (i.e., canals, rivers, and parks) in twelve districts of Ho Chi Minh City and tested against a library of 12 antibiotic-resistant genes and 1 integron gene. The susceptibility of isolated Escherichia coli from sludge and soil (n = 104 and 129, respectively) was tested against nine antibiotics. Over 60% of sludge and soil samples harbored sul1, ere(A), intI1, cmIA, and tet(A) genes. The three most common phenotypic resistances found in E. coli isolated from sludge and soil were to ampicillin, tetracycline, and sulfamethoxazole/trimethoprim. In a temporal microcosm study of antibiotic-susceptible and multi-drug-resistant E. coli inoculated in soil, temperature (21.4 vs. 30 °C), resistance phenotype, and soil background microbial community were associated with E. coli decay rates over 73 days. This is the first study that provides insights into the high prevalence of antibiotic resistance in septic systems and environmental reservoirs in Ho Chi Minh City, Vietnam. Findings highlight that the fecal sludge and soil environments in Vietnam are likely reservoirs for dissemination of and human exposure to antibiotic resistance.201931643014
2857120.9998Changes in antibiotic resistance of Escherichia coli during the broiler feeding cycle. The purpose of this study was to investigate the drug-resistant phenotypes and genes of Escherichia coli in animal, environmental, and human samples before and after antibiotic use at a large-scale broiler farm to understand the respective effects on E. coli resistance during the broiler feeding cycle. The antibiotic use per broiler house was 143.04 to 183.50 mg/kg, and included tilmicosin, florfenicol, apramycin, and neomycin. All strains isolated on the first day the broilers arrived (T1; day 1) were antibiotic-resistant bacteria. E. coli strains isolated from animal samples were resistant to ampicillin, tetracycline, and sulfamethoxazole (100%), and those isolated from environmental samples were resistant to 5 different drugs (74.07%, 20 of 27). E. coli strains isolated on the last day before the broilers left (T2; day 47) had a higher resistance rate to florfenicol (100%, 36 of 36) than at T1 (P < 0.05). Multidrug resistance increased from T1 (84.21%, 32 of 38) to T2 (97.22%, 35 of 36). Most strains were resistant to 5 classes of antibiotics, and 2 strains were resistant to 6 classes of antibiotics. Among 13 identified drug resistance genes, 11 and 13 were detected at T1 and T2, respectively. NDM-1 was detected in 4 environmental samples and 1 animal sample. In conclusion, the use of antibiotics during breeding increases E. coli resistance to antibacterial drugs. Drug-resistant bacteria in animals and the environment proliferate during the feeding cycle, leading to the widespread distribution of drug resistance genes and an increase in the overall resistance of bacteria.202033248614
5269130.9998Prevalence of antibiotic resistance genes in bacteria from Gomti and Ganga rivers: implications for water quality and public health. Rivers serve as a significant habitat and water sources for diverse organisms, including humans. An important environmental and public health concern is the increase in antibiotic-resistant bacteria (ARBs) and genes (ARGs) in aquatic ecosystems brought about by excessive pollutant flow. The research highlighted that river water, which is receiving discharge from wastewater treatment plants, is harbouring multidrug-resistant bacteria. River water samples were collected in January, April, July and October 2022 from three separate locations of each Gomti and Ganga river. A total of 114 bacteria were isolated from Gomti as well as the Ganga River. All the isolates were tested for their resistance to various antibiotics by disc diffusion method. The isolated bacteria were tested for the antibiotic resistance genes using PCR and were identified by 16s rRNA sequencing. The ARBs percentages for each antibiotic were as follows: ampicillin (100%); cefotaxime (96.4, 63.1%); erythromycin (52.6, 57.8%); amikacin (68.4, 50.8%); tetracycline (47.3, 54.3%); nalidixic acid (47.3, 45.6%); streptomycin (68.4, 49.1%); gentamycin (43.8, 35%); chloramphenicol (26.3, 33.3%); neomycin (49.1, 29.8%) and ciprofloxacin (24.5, 7.01%). Further, antibiotic resistance genes in Gomti and Ganga water samples disclose distinctive patterns, including resistance to ermB (25, 40%); tetM (25, 33.3%); ampC (44.4, 40%) and cmlA1 (16.6%). Notably cmlA1 resistant genes were absent in all bacterial strains of the Gomti River. Additionally, gyrA gene was not found in both the river water samples. The presence of ARGs in the bacteria from river water shows threat of transferring these genes to native environmental bacteria. To protect the environment and public health, constant research is necessary to fully understand the extent and consequences of antibiotic resistance in these aquatic habitats.202439349711
2854140.9998Occurrence of antibiotic resistance genes in culturable bacteria isolated from Turkish trout farms and their local aquatic environment. Antibiotic resistance and presence of the resistance genes were investigated in the bacteria isolated from water, sediment, and fish in trout farms. A total of 9 bacterial species, particularly Escherichia coli, were isolated from the water and sediment samples, and 12 species were isolated from fish. The antimicrobial test indicated the highest resistance against sulfamethoxazole and ampicillin in coliform bacteria, and against sulfamethoxazole, imipenem, and aztreonam in known pathogenic bacteria isolated from fish. The most effective antibiotics were rifampicin, chloramphenicol, and tetracycline. The multiple antibiotic resistance index was above the critical limit for almost all of the bacteria isolated. The most common antibiotic resistance gene was ampC, followed by tetA, sul2, blaCTX-M1, and blaTEM in the coliform bacteria. At least one resistance gene was found in 70.8% of the bacteria, and 66.6% of the bacteria had 2 or more resistance genes. Approximately 36.54% of the bacteria that contain plasmids were able to transfer them to other bacteria. The plasmid-mediated transferable resistance genes were ampC, blaCTX-M1, tetA, sul2, and blaTEM. These results indicate that the aquatic environment could play an important role in the development of antibiotic resistance and the dissemination of resistance genes among bacteria.201525993887
5263150.9998Seasonal Variations in Water-Quality, Antibiotic Residues, Resistant Bacteria and Antibiotic Resistance Genes of Escherichia coli Isolates from Water and Sediments of the Kshipra River in Central India. OBJECTIVES: To characterize the seasonal variation, over one year, in water-quality, antibiotic residue levels, antibiotic resistance genes and antibiotic resistance in Escherichia coli isolates from water and sediment of the Kshipra River in Central India. METHODS: Water and sediment samples were collected from seven selected points from the Kshipra River in the Indian city of Ujjain in the summer, rainy season, autumn and winter seasons in 2014. Water quality parameters (physical, chemical and microbiological) were analyzed using standard methods. High-performance liquid chromatography⁻tandem mass spectrometry was used to determine the concentrations of antibiotic residues. In river water and sediment samples, antibiotic resistance and multidrug resistance patterns of isolated E. coli to 17 antibiotics were tested and genes coding for resistance and phylogenetic groups were detected using multiplex polymerase chain reaction. One-way analysis of variance (ANOVA) and Fisher tests were applied to determine seasonal variation. RESULTS: In river water, seasonal variation was significantly associated with various water quality parameters, presence of sulfamethoxazole residues, bacteria resistant to ampicillin, cefepime, meropenem, amikacin, gentamicin, tigecycline, multidrug resistance and CTX-M-1 gene. The majority of the Extended Spectrum Beta-Lactamase (ESBL)-producing E. coli isolates from river water and sediment in all different seasons belonged to phylogenetic group A or B1. CONCLUSIONS: Antibiotic pollution, resistance and resistance genes in the Kshipra River showed significant seasonal variation. Guidelines and regulatory standards are needed to control environmental dissemination of these “pollutants” in this holy river.201829914198
5264160.9998Comparison of Culture- and Quantitative PCR-Based Indicators of Antibiotic Resistance in Wastewater, Recycled Water, and Tap Water. Standardized methods are needed to support monitoring of antibiotic resistance in environmental samples. Culture-based methods target species of human-health relevance, while the direct quantification of antibiotic resistance genes (ARGs) measures the antibiotic resistance potential in the microbial community. This study compared measurements of tetracycline-, sulphonamide-, and cefotaxime-resistant presumptive total and fecal coliforms and presumptive enterococci versus a suite of ARGs quantified by quantitative polymerase chain reaction (qPCR) across waste-, recycled-, tap-, and freshwater. Cross-laboratory comparison of results involved measurements on samples collected and analysed in the US and Portugal. The same DNA extracts analysed in the US and Portugal produced comparable qPCR results (variation <28%), except for bla(OXA-1) gene (0%-57%). Presumptive total and fecal coliforms and cefotaxime-resistant total coliforms strongly correlated with bla(CTX-M) and intI1 (0.725 ≤ R(2) ≤ 0.762; p < 0.0001). Further, presumptive total and fecal coliforms correlated with the Escherichia coli-specific biomarkers, gadAB, and uidA, suggesting that both methods captured fecal-sourced bacteria. The genes encoding resistance to sulphonamides (sul1 and sul2) were the most abundant, followed by genes encoding resistance to tetracyclines (tet(A) and tet(O)) and β-lactams (bla(OXA-1) and(,)bla(CTX-M)), which was in agreement with the culture-based enumerations. The findings can help inform future application of methods being considered for international antibiotic resistance surveillance in the environment.201931671709
5257170.9998Removal of fecal indicator bacteria and antibiotic resistant genes in constructed wetlands. Wastewater discharge evidently increased bacterial diversity in the receiving waterbodies. The objective of this study was to evaluate the effectiveness of a constructed wetland in reducing fecal indicator bacteria (FIB) and antibiotic resistant genes (ARGs). We determined the prevalence and attenuation of fecal indicator bacteria including Escherichia coli and enterococci, along with ARGs, and human-associated Bacteroidales (HF183) markers by quantitative polymerase chain reaction (qPCR) method. Three types of water samples (inlet, intermediate, and outlet) from a constructed wetland were collected once a month from May to December in 2013. The overall reduction of E. coli was 50.0% based on culture method. According to the qPCR result, the overall removal rate of E. coli was only 6.7%. Enterococci were found in 62.5% of the wetland samples. HF183 genetic marker was detected in all final effluent samples with concentration ranging from 1.8 to 4.22 log(10) gene copies (GC)/100 ml. Of the ARGs tested, erythromycin resistance genes (ermF) were detected in 79.2% of the wetland samples. The class 1 integrase (intI1) was detected in all water samples with concentration ranging from 0.83 to 5.54 log(10) GC/100 ml. The overall removal rates of enterococci, HF183, intI1, and ermF were 84.0%, 66.6%, 67.2%, and 13.1%, respectively.201930758793
2772180.9998Antibiotic Resistance in Pseudomonas spp. Through the Urban Water Cycle. Selection and dissemination of resistant bacteria and antibiotic resistance genes (ARGs) require a deeper understanding since antibiotics are permanently released to the environment. The objective of this paper was to evaluate the phenotypic resistance of 499 isolates of Pseudomonas spp. from urban water sources, and the prevalence of 20 ARGs within those isolates. Resistance to penicillins, cephalosporins, carbapenems, quinolones, macrolides, and tetracyclines was mainly observed in the hospital effluent, municipal wastewater and river water downstream the city. Resistant strains were frequently identified as P. aeruginosa and P. putida. P. aeruginosa isolates were mostly resistant to cefepime, ceftazidime, imipenem, and gentamycin, while P. putida strains were especially resistant to piperacillin-tazobactam. ARGs such as bla(TEM-1), bla(SHV-1), bla(PER-1), bla(AmpC), bla(VIM-1), PstS, qnrA, qnrB, ermB, tetA, tetB and tetC have been detected. The bla(AmpC) gene was found in P. aeruginosa, while bla(TEM-1) and bla(PER-1) genes were found in P. putida. Class 1 integron integrase gene was found in 6.81% of the Pseudomonas isolates.202133625570
2862190.9998Regulation Transcriptional of Antibiotic Resistance Genes (ARGs) in Bacteria Isolated from WWTP. The incidence of antibiotics and transcriptional regulation of ARGs in isolated bacteria from wastewater needs to be explored. By HPLC, in samples of untreated wastewater, ampicillin (49.74 ± 5.70 µg/mL), chloramphenicol (0.60 ± 0.03 µg/mL), tylosin (72.95 ± 2.03 µg/mL), and oxytetracycline (0.22 ± 0.01 µg/mL) was determined. Through metagenomic analysis identified 58 bacterial species belonging to 9 phyla and at least 14 species have shown resistance to a variety of antibiotics. Twenty-two bacterial isolates were proved to be resistant to fifteen antibiotics of new generation and used in medical research to combat infectious diseases. Fourteen strains were shown to harbor plasmids in size ranges of 2-5 Kb, 6-10 Kb and plasmids with size greater than 10 Kb. By quantitative PCR it was possible to identify genes sul, qnr, cat1, aadA1, and sat-1 gene were shown to be present in gDNA samples from treated and untreated samples of wastewater and by relative expression analysis, differential expression of cat1, ermB, act, and tetA genes was demonstrated in strains that showed identity with Escherichia coli, Bacteroides fragilis, and Salmonella thyphi, and that were stressed with different concentrations of antibiotics. The presence of ARGs in untreated water samples, as well as in bacterial isolates, was indicative that in these habitats there are microorganisms that can resist β-lactams, aminoglycosides, tetracyclines, sulfonamides, and quinolones.202337672120