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227200.9964Routine wastewater-based monitoring of antibiotic resistance in two Finnish hospitals: focus on carbapenem resistance genes and genes associated with bacteria causing hospital-acquired infections. BACKGROUND: Wastewater-based monitoring represents a useful tool for antibiotic resistance surveillance. AIM: To investigate the prevalence and abundance of antibiotic resistance genes (ARGs) in hospital wastewater over time. METHODS: Wastewater from two hospitals in Finland (HUS1 and HUS2) was monitored weekly for nine weeks (weeks 25-33) in summer 2020. A high-throughput real-time polymerization chain reaction (HT-qPCR) system was used to detect and quantify 216 ARGs and genes associated with mobile genetic elements (MGEs), integrons, and bacteria causing hospital-acquired infections (HAIs), as well as the 16S rRNA gene. Data from HT-qPCR were analysed and visualized using a novel digital platform, ResistApp. Eight carbapenem resistance genes (blaGES, blaKPC, blaVIM, blaNDM, blaCMY, blaMOX, blaOXA48, and blaOXA51) and three genes associated with bacteria causing HAIs (Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa) were studied. FINDINGS: There was a significantly higher number of ARGs at both hospitals in weeks 27-30 (174-191 genes) compared to other sampling weeks (151-171 genes). Our analyses also indicated that the two hospitals, which used different amounts of antibiotics, had significantly different resistance gene profiles. Carbapenem resistance genes were more prevalent and abundant in HUS1 than HUS2. Across both hospitals, blaGES and blaVIM were the most prevalent and abundant. There was also a strong positive association between blaKPC and K. pneumoniae in HUS1 wastewater. CONCLUSION: Routine wastewater-based monitoring using ResistApp can provide valuable information on the prevalence and abundance of ARGs in hospitals. This helps hospitals understand the spread of antibiotic resistance in hospitals and identify potential areas for intervention.202134537275
526310.9962Seasonal 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
349120.9961Assessment of seasonal variations in antibiotic resistance genes and microbial communities in sewage treatment plants for public health monitoring. The spread of antimicrobial resistance (AMR) around the globe, especially in the urban cities with high population, is a major concern. Therefore, the current study aims at identifying antibiotic resistant bacteria, microbial community compositions and the quantification of antimicrobial resistant genes from six sewage treatment plants (STPs) across Pune city in Maharashtra, India. A total of 106 isolates obtained were tested against six antibiotics in which the highest resistance was observed against trimethoprim (24.53 %). The qPCR assays of seven antibiotic resistance genes revealed abundance of bla(imp-1) and mecA genes in the summer and monsoon seasons followed by bla(NDM-1) gene in the summer and winter seasons. The alpha diversity indices depicted highest microbial diversity of inlet samples during winter, followed by inlet samples during the summer and monsoon seasons. Comparative analysis revealed Bifidobacterium (51 %), Pseudomonas (28.7 %) and Zoogloea (17.6 %) as the most abundant genera in the inlet samples during the summer, monsoon and winter seasons respectively while Acinetobacter (31 %) and Flavobacterium (23 % in winter and 18.2 % in summer) dominated the outlet samples. The co-network analysis revealed positive and negative interactions in the winter and monsoon but only positive interactions in the summer season. Venn diagrams showed higher abundance of ASVs in the outlet samples than the inlet. The top genera correlated exactly opposite with the pH compared to BOD and COD. PICRUSt2-based functional prediction revealed a higher abundance of methicillin resistance, β-lactamase resistance and multidrug resistance genes in inlet samples while chloramphenicol resistance was found higher in outlet samples. Further, we observed that potential pathogens causing infectious disease such as pertussis, shigellosis and tuberculosis were present in all three seasons.202540320120
526730.9961Diversity and antibiotic resistance of cultivable bacteria in bulk tank milk from dairy farms in Shandong Province, China. INTRODUCTION: This study systematically analyzed bacterial diversity and antimicrobial resistance (AMR) profiles in bulk tank milk from five dairy farms (n = 30) in Shandong Province, China, to assess public health risks associated with microbial contamination and provide critical data for regional quality control and AMR risk assessment in dairy production systems. METHODS: Total bacterial counts were quantified, revealing significant inter-farm variation (P < 0.05) with a range of 3.94-6.68 log CFU/mL. Among 129 bacterial isolates, genus-level dominance and species prevalence were identified. Antimicrobial susceptibility testing (AST) against 10 agents was performed using integrated resistance criteria combining Clinical and Laboratory Standards Institute (CLSI) standards and epidemiological cutoff values (ECOFFs). Nine resistance genes targeting seven antibiotic classes were detected via PCR. RESULTS: The highest resistance rate was observed for sulfadiazine (53.2%) and the lowest for levofloxacin (6.0%). Multidrug resistance was detected in 23% (20/87) of isolates, with 14 strains meeting ECOFFs-based resistance criteria. PCR analysis showed sul1 (70.5%) and ant(4')-Ia (54.3%) as the most prevalent resistance genes, while mcr-1, lnu (B), and bla (NDM-1) were absent in all isolates. Regional resistance variations correlated significantly with farm management practices. DISCUSSION: These findings underscore the impact of historical antibiotic use on AMR dissemination. Enhanced AMR surveillance in raw milk, improved antibiotic stewardship, and targeted interventions are crucial to mitigate public health risks from microbial contamination and horizontal gene transfer of resistance determinants.202540771950
287740.9961Metagenomic insights into isolable bacterial communities and antimicrobial resistance in airborne dust from pig farms. This study aims to investigate bacterial communities and antimicrobial resistance (AMR) in airborne dust from pig farms. Airborne dust, pig feces and feed were collected from nine pig farms in Thailand. Airborne dust samples were collected from upwind and downwind (25 meters from pig house), and inside (in the middle of the pig house) of the selected pig house. Pig feces and feed samples were individually collected from the pen floor and feed trough from the same pig house where airborne dust was collected. A direct total bacteria count on each sampling plate was conducted and averaged. The ESKAPE pathogens together with Escherichia coli, Salmonella, and Streptococcus were examined. A total of 163 bacterial isolates were collected and tested for MICs. Pooled bacteria from the inside airborne dust samples were analyzed using Metagenomic Sequencing. The highest bacterial concentration (1.9-11.2 × 10(3) CFU/m(3)) was found inside pig houses. Staphylococcus (n = 37) and Enterococcus (n = 36) were most frequent bacterial species. Salmonella (n = 3) were exclusively isolated from feed and feces. Target bacteria showed a variety of resistance phenotypes, and the same bacterial species with the same resistance phenotype were found in airborne dust, feed and fecal from each farm. Metagenomic Sequencing analysis revealed 1,652 bacterial species across all pig farms, of which the predominant bacterial phylum was Bacillota. One hundred fifty-nine AMR genes of 12 different antibiotic classes were identified, with aminoglycoside resistance genes (24%) being the most prevalent. A total of 251 different plasmids were discovered, and the same plasmid was detected in multiple farms. In conclusion, the phenotypic and metagenomic results demonstrated that airborne dust from pig farms contained a diverse array of bacterial species and genes encoding resistance to a range of clinically important antimicrobial agents, indicating the significant role in the spread of AMR bacterial pathogens with potential hazards to human health. Policy measurements to address AMR in airborne dust from livestock farms are mandatory.202438872793
526150.9961Prevalence of antibiotic resistance genes from effluent of coastal aquaculture, South Korea. The wide use of antibiotics in aquaculture for prophylactic and therapeutic purposes can potentially lead to the prevalence of antibiotic resistance genes (ARGs). This study reports for the first time the profile of ARGs from effluents of coastal aquaculture located in South Jeolla province and Jeju Island, South Korea. Using quantitative PCR (qPCR), twenty-two ARGs encoding tetracycline resistance (tetA, tetB, tetD, tetE, tetG, tetH, tetM, tetQ, tetX, tetZ, tetBP), sulfonamide resistance (sul1, sul2), quinolone resistance (qnrD, qnrS, aac(6')-Ib-cr), β-lactams resistance (bla(TEM), bla(CTX), bla(SHV)), macrolide resistance (ermC), florfenicol resistance (floR) and multidrug resistance (oqxA) and a class 1 integrons-integrase gene (intI1) were quantified. In addition, Illumina Miseq sequencing was applied to investigate microbial community differences across fish farm effluents. Results from qPCR showed that the total number of detected ARGs ranged from 4.24 × 10(-3) to 1.46 × 10(-2) copies/16S rRNA gene. Among them, tetB and tetD were predominant, accounting for 74.8%-98.0% of the total ARGs. Furthermore, intI1 gene showed positive correlation with tetB, tetD, tetE, tetH, tetX, tetZ tetQ and sul1. Microbial community analysis revealed potential host bacteria for ARGs and intI1. Two genera, Vibrio and Marinomonas belonging to Gammaproteobacteria, showed significant correlation with tetB and tetD, the most dominant ARGs in all samples. Also, operational taxonomic units (OTUs)-based network analysis revealed that ten OTUs, classified into the phyla Proteobacteria, Cyanobacteria/Chloroplast, Bacteroidetes, Verrucomicrobia and an unclassified phylum, were potential hosts of tetracycline resistance genes (i.e., tetA, tetG, tetH, tetM, tetQ and tetZ). Further systematic monitoring of ARGs is warranted for risk assessment and management of antibacterial resistance from fish farm effluents.201829031406
258460.9960Resistance to medically important antimicrobials in broiler and layer farms in Cameroon and its relation with biosecurity and antimicrobial use. INTRODUCTION: Poultry production accounts for 42% of Cameroonian meat production. However, infectious diseases represent the main hindrance in this sector, resulting in overuse and misuse of antimicrobials that can contribute to the emergence and dissemination of antimicrobial resistance (AMR). This study aimed to evaluate the prevalence of antimicrobial resistance genes (ARGs) conferring resistance to carbapenems (bla(VIM-2) and bla(NDM) ), (fluoro) quinolones (qnrS, qnrA, and qnrB), polymyxins (mcr1 to mcr5), and macrolides (ermA and ermB) in the poultry farm environment. Additionally, the study examined the relationship between these ARGs and biosecurity implementation, as well as farmers' knowledge, attitudes, and practices toward antimicrobial use (AMU) and AMR, including their perception of AMR risk. MATERIALS AND METHODS: Fecal, drinking water, and biofilm samples from drinking water pipelines were collected from 15 poultry farms and subsequently analyzed by real-time PCR and 16S rRNA NGS. RESULTS: All samples tested positive for genes conferring resistance to (fluoro) quinolones, 97.8% to macrolides, 64.4% to polymyxins, and 11.1% to carbapenems. Of concern, more than half of the samples (64.4%) showed a multi-drug resistance (MDR) pattern (i.e., resistance to ≥3 antimicrobial classes). Drinking water and biofilm microbial communities significantly differed from the one of the fecal samples, both in term of diversity (α-diversity) and composition (β-diversity). Furthermore, opportunistic pathogens (i.e., Comamonadaceae and Sphingomonadaceae) were among the most abundant bacteria in drinking water and biofilm. The level of biosecurity implementation was intermediate, while the knowledge and attitude of poultry farmers toward AMU were insufficient and unsuitable, respectively. Good practices toward AMU were found to be correlated with a reduction in polymyxins and MDR. DISCUSSION: This study provides valuable information on resistance to medically important antimicrobials in poultry production in Cameroon and highlights their potential impact on human and environmental health.202439881983
272570.9960Hygiene practices and antibiotic resistance among dental and medical students: a comparative study. PURPOSE: Healthcare students' hand and smartphone hygiene is critical due to potential pathogenic and antibiotic-resistant bacteria transmission. This study evaluates hygiene practices in medical and dental students at Kuwait University, exploring antibiotic resistance gene prevalence. METHODS: Swab samples were collected from the hands and smartphones of 32 medical and 30 dental students. These samples were cultured on Columbia Blood Agar and McConkey Agar plates to quantify bacterial colony-forming units (CFUs). The extracted DNA from these colonies underwent RT-PCR to identify antibiotic resistance genes, including tem-1, shv, blaZ, and mecA. Additionally, a questionnaire addressing hygiene practices was distributed post-sample collection. RESULTS: Medical students exhibited more frequent hand hygiene compared to dental students (P ≤ 0.0001). Although significantly fewer bacterial CFUs were found on medical students' smartphones (mean = 35 ± 53) than dental students' (mean = 89 ± 129) (P ≤ 0.05), no significant differences were observed in CFU counts on their hands (medical: mean = 17 ± 37; dental: mean = 96 ± 229). Detection of at least one of the targeted antibiotic resistance genes on medical (89% hands, 52% smartphones) and dental students' (79% hands, 63% smartphones) was not statistically significant. However, the prevalence of two genes, tem-1 and shv, was significantly higher on medical students' hands (78% and 65%, respectively) than on dental students' hands (32% and 28%, respectively). CONCLUSION: Clinically significant prevalence of antibiotic resistance genes were found on medical and dental students' hands and smartphones, emphasizing the importance of ongoing education regarding hand hygiene and smartphone disinfection. This continuous reinforcement in the curriculum is crucial to minimizing the risk of cross-contamination.202438514584
349480.9960Pathogenic bacteria and antibiotic resistance genes in hospital indoor bioaerosols: pollution characteristics, interrelation analysis, and inhalation risk assessment. Hospitals are high risk areas for the spread of diseases, with indoor bioaerosols containing a variety of pathogens. Inhalation of these pathogens may cause severe nosocomial infections in patients and medical staff. A comprehensive investigation was conducted during the influenza A outbreak to explore the distribution and pathogenic risk of airborne pathogens and antibiotic resistance genes (ARGs) across different hospital departments. It was revealed that airborne bacterial concentrations ranged from 118 to 259 CFU/m(3), and the main aerosol particle size was 4.7-5.8 μm (27.7 %). The proportion of bioaerosols smaller than 2.5 μm was highest in the respiratory waiting area (59.3 %). The dominant pathogens detected in hospital air were Bacillus, Staphylococcus, Pseudomonas and Micrococcus. The absolute abundance of ARGs/mobile genetic elements (MGEs) ranged from 0.55 to 479.44 copies/m(3), peaking in the respiratory ward air. TetL-02, lnuA-01, intI1, ermB, and qacEdelta1-02 were the top five ARGs/MGEs in hospital air. Moreover, doctors inhaled higher doses of ARGs/MGEs in inpatient wards than outpatient waiting areas. Network analysis identified Pseudomonas, Micrococcus, Microbacterium, and Enterobacter as potential ARGs reservoirs. The Bugbase result showed the presence of potentially pathogenic pathogens in the bioaerosols at all sampling sites. The quantitative microbiological risk assessment results further showed that airborne Staphylococcus could pose an infection risk to medical staff. It was determined that the use of masks was effective in reducing this risk to an acceptable level. This study will provide a scientific basis for comprehensively understanding the characteristics and potential risks of hospital bioaerosols during the outbreak of respiratory infectious diseases.202540222613
209990.9960Antibiotic-resistant bacteria and resistance-genes in drinking water source in north Shoa zone, Amhara region, Ethiopia. BACKGROUND: The growing number of antimicrobial-resistant bacteria in a range of environments poses a serious challenge to infectious disease prevention. Good water quality is critical to human health and has a direct impact on a country's socio-economic growth. Therefore, assessing the bacteriological quality of drinking water provides benchmark data and provides insight into the development of further protection and treatment measures. METHODS: A cross-sectional study was conducted from February 1, 2022, to September 31, 2023, in the diarrhea hotspot areas of North Shewa Zone (Minjar-Shenkora and Mojana-Wedera districts). Water samples were collected from drinking water sources (hand-pumps, boreholes, wells, spring water and ponds) to assess the quality following WHO guidelines. The collected water samples were processed for bacterial isolation, antimicrobial susceptibility testing, and detection of antimicrobial resistance genes. Data were entered and analyzed using the Statistical Package for the Social Sciences (SPSS) version 25. RESULTS: A total of (49/138, 35.5%) bacteria were isolated from 138 drinking water samples, with a positive rate of (41/138, 29.7%). Among the isolates, (16/138, 11.6%) were Staphylococcus aureus while (33/138, 23.9%) were members of Enterobacteriaceae. Relatively high resistance rate among all isolates were observed for the most prescribed antibiotics in Ethiopia, including erythromycin, cotrimoxazole, doxycycline, ceftriaxone, gentamicin, and chloramphenicol. However, a low resistance was observed for early introduced antibiotics such as ciprofloxacin and recently introduced antibiotics such as cefotaxime, ceftazidime, imipenem, and meropenem. Among the 49 bacteria isolates, (32/49, 65.3%) were multidrug-resistant (MDR) pathogens while (12/49, 24.5%) were ESβL producers. Different ESβL genes were detected in most bacterial isolates. The predominant ESβL genes were blaCTX-M-gp8/25 (6/33, 18.2%), blaCTX-M-gp9 (5/33, 15.2%), and blaCTX-M-gp1 (5/33, 15.2%). CONCLUSION: The result of this study suggests that most water sources in the study area were contaminated by various bacterial species that are resistant to different antibiotics. Various ESβL resistance genes have also been detected. Therefore, regular sanitary inspection and bacteriological analysis should be mandatory to protect drinking water sources from contamination and the persistence of resistant bacteria.202439310913
5242100.9959Highly sensitive detection of antimicrobial resistance genes in hospital wastewater using the multiplex hybrid capture target enrichment. Wastewater can be useful in monitoring the spread of antimicrobial resistance (AMR) within a hospital. The abundance of antibiotic resistance genes (ARGs) in hospital effluent was assessed using metagenomic sequencing (mDNA-seq) and hybrid capture (xHYB). mDNA-seq analysis and subsequent xHYB targeted enrichment were conducted on two effluent samples per month from November 2018 to May 2021. Reads per kilobase per million (RPKM) values were calculated for all 1,272 ARGs in the constructed database. The monthly numbers of patients with presumed extended-spectrum β-lactamase (ESBL)-producing and metallo-β-lactamase (MBL)-producing bacteria, methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant enterococci (VRE) were compared with the monthly RPKM values of bla(CTX-M), bla(IMP), mecA, vanA, and vanB by xHYB. The average RPKM value for all ARGs detected by xHYB was significantly higher than that of mDNA-seq (665, 225, and 328, respectively, and P < 0.05). The average number of patients with ESBL producers and RPKM values of bla(CTX-M-1) genes in 2020 were significantly higher than that in 2019 (17 and 13 patients per month and 921 vs 232 per month, respectively, both P < 0.05). The average numbers of patients with MBL-producers, MRSA, and VRE were 1, 28, and 0 per month, respectively, while the average RPKM values of bla(IMP), mecA, vanA, and vanB were 6,163, 6, 0, and 126 per month, respectively. Monitoring ARGs in hospital effluent using xHYB was found to be more useful than conventional mDNA-seq in detecting ARGs including bla(CTX-M), bla(IMP,) and vanB, which are important for infection control.IMPORTANCEEnvironmental ARGs play a crucial role in the emergence and spread of AMR that constitutes a significant global health threat. One major source of ARGs is effluent from healthcare facilities, where patients are frequently administered antimicrobials. Culture-independent methods, including metagenomics, can detect environmental ARGs carried by non-culturable bacteria and extracellular ARGs. mDNA-seq is one of the most comprehensive methods for environmental ARG surveillance; however, its sensitivity is insufficient for wastewater surveillance. This study demonstrates that xHYB appropriately monitors ARGs in hospital effluent for sensitive identification of nosocomial AMR dissemination. Correlations were observed between the numbers of inpatients with antibiotic-resistant bacteria and the ARG RPKM values in hospital effluent over time. ARG surveillance in hospital effluent using the highly sensitive and specific xHYB method could improve our understanding of the emergence and spread of AMR within a hospital.202337222510
7781110.9959Untreated HWWs Emerged as Hotpots for ARGs. Hospital wastewaters (HWWs) are reported to be hotspots for antibiotics and antibiotic-resistant bacteria. However, limited information involves the impact of these effluents on dissemination of antibiotic-resistance genes (ARGs). In this study, therefore, seasonally collected HWWs were monitored for overall bacterial load and seven ARGs aadA, tetA, cmlA, sul1, qnrS, ermB and bla (CTX-M )by using quantitative polymerase chain reaction method. Overall bacterial 16S rRNA copy number was found to be the lowest in winter with 10(3 )copy number/mL, while the highest copy number, with 10(5 )copy number/mL, was observed in both summer and spring. All hospitals tested displayed similar seasonal ARG copy number profile of aadA > tetA > cmlA ≈ sul1 > ermB ≈ qnrS > bla (CTX-M). The results indicated that untreated HWWs were hotspots for ARGs and required attention before discharging into public sewer.202031965225
2727120.9959Prevalence and Antibiotic Resistance Pattern of Streptococcus, Staphylococcus, Neisseria meningitidis and Enterobacteriaceae in Two Reference Hospitals of Yaoundé: An Overview before and during COVID-19 Pandemic Era. The COVID-19 pandemic led to tremendously use of antimicrobial due to the lack of proper treatment strategies, raising concerns about emergence of antimicrobial resistance (AMR). This study aimed at determining the prevalence and antibiotic resistance pattern of selected bacteria isolates in 02 referral health facilities in Yaoundé before and during the COVID-19 pandemic era. We conducted a retrospective study over a period of 03 years (from 1 January 2019 to 31 December 2021) in the bacteriology units of the Central and General Hospitals of Yaoundé, Cameroon. Data on bacteria genera (Streptococcus, Staphylococcus, Neisseria meningitidis and Enterobacteriaceae) as well as their corresponding specifics antibiotics: Cefixime, azythromycin and erythromycin were obtained from laboratory records. The global resistance rate of bacteria as well as their correlation with antibiotics according to COVID-19 pandemic era was determined and compared. For p < 0.05, the difference was statistically significant. In all, 426 bacterial strains were included. It appeared that the highest number of bacteria isolates and lowest rate of bacterial resistance were recorded during the pre-COVID-19 period in 2019 (160 isolates vs. 58.8% resistance rate). Conversely, lower bacteria strains but greater resistance burden were recorded during the pandemic era (2020 and 2021) with the lowest bacteria amount and peak of bacteria resistance registered in 2020, the year of COVID-19 onset (120 isolates vs. 70% resistance in 2020 and 146 isolates vs. 58.9% resistance in 2021). In contrast to almost all others groups of bacteria where the resistance burden was quite constant or decreasing over years, the Enterobacteriaceae exhibited greater resistance rate during the pandemic period [60% (48/80) in 2019 to 86.9% (60/69) in 2020 and 64.5% (61/95) in 2021)]. Concerning antibiotics, unlike erythromycin, azythromycin related resitance increased during the pandemic period and the resistance to Cefixim tends to decrease the year of the pandemic onset (2020) and re-increase one year therafter. A significant association was found between resistant Enterobacteriaceae strains and cefixime (R = 0.7; p = 0.0001) and also, between resistant Staphylococcus strains and erythromycin (R = 0.8; p = 0.0001). These retrospective data showed a herogeneous MDR bacteria rate and antibiotic resistance pattern over time before and during the COVID-19 pandemic era suggesting that antimicrobial resistance needs to be more closely monitored.202337237832
3113130.9959Resistome in the indoor dust samples from workplaces and households: a pilot study. The antibiotic resistance genes (ARGs) limit the susceptibility of bacteria to antimicrobials, representing a problem of high importance. Current research on the presence of ARGs in microorganisms focuses mainly on humans, livestock, hospitals, or wastewater. However, the spectrum of ARGs in the dust resistome in workplaces and households has gone relatively unexplored. This pilot study aimed to analyze resistome in indoor dust samples from participants' workplaces (a pediatric hospital, a maternity hospital, and a research center) and households and compare two different approaches to the ARGs analysis; high-throughput quantitative PCR (HT-qPCR) and whole metagenome shotgun sequencing (WMGS). In total, 143 ARGs were detected using HT-qPCR, with ARGs associated with the macrolides, lincosamides, and streptogramin B (MLSB) phenotype being the most abundant, followed by MDR (multi-drug resistance) genes, and genes conferring resistance to aminoglycosides. A higher overall relative quantity of ARGs was observed in indoor dust samples from workplaces than from households, with the pediatric hospital being associated with the highest relative quantity of ARGs. WMGS analysis revealed 36 ARGs, of which five were detected by both HT-qPCR and WMGS techniques. Accordingly, the efficacy of the WMGS approach to detect ARGs was lower than that of HT-qPCR. In summary, our pilot data revealed that indoor dust in buildings where people spend most of their time (workplaces, households) can be a significant source of antimicrobial-resistant microorganisms, which may potentially pose a health risk to both humans and animals.202439691696
5264140.9958Comparison 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
2712150.9958Antibiotic Resistance Profiles of Bacteria Isolated from Hotspot Environments in Bahir Dar City, Northwestern Ethiopia. BACKGROUND: Wastes generated from hotspot environments contain a wide range of antibiotics and pathogens that play a significant role in the dissemination of antibiotic-resistant bacteria in the environment. This study was carried out to isolate bacteria from hotspot environments and determine their resistance profiles to commonly used antibiotics in Bahir Dar city, Ethiopia. METHODS: A cross-sectional study was conducted from October 2020 to June 2021 in Bahir Dar City. A total of 126 waste and wastewater samples were aseptically collected, transported, and processed for bacteriological isolation and susceptibility testing following standard procedures. RESULTS: A total of 411 bacterial isolates were recovered and the highest value of 122 (29.7%) bacterial isolates were obtained from medical wastewater samples, and the most frequently isolated bacteria were assigned to the species Escherichia coli with 82 strains (19.5%). The results revealed that the highest resistance profile of 69 (95.8%) was obtained in Staphylococcus aureus against ampicillin and 46 (86.8%) Citrobacter spp. against tetracycline. Two hundred and sixteen (52.6%) of bacteria showed multi-drug resistance and the highest multi-drug resistance was observed in Pseudomonas spp. 47 (65.3%), followed by Escherichia coli 51 (62.2%). The highest resistance profile of 12 (85.7%) and 60 (74.1%) for tetracycline were obtained from beef waste and wastewater and medical wastewater samples. The highest multi-drug resistance was recorded in isolates isolated from beef waste and wastewater samples 11 (64.7%), followed by medical wastewater samples 84 (64.1%). Even though a higher (>0.2) multi-antibiotic resistance index was found in all hotspot environments, the highest multi-antibiotic resistance index (0.477) was recorded in bacteria isolated from medical wastewater. CONCLUSION: It was concluded that wastes generated from hotspot environments and released in the environment contain large numbers of antibiotic-resistant, multidrug, extensively, and pan-drug-resistant bacteria. Proper waste management strategies should be established.202235785260
2720160.9958Phenotypic and genotypic characterization of antimicrobial resistance in Enterococcus spp. Isolated from the skin microbiota of channel catfish (Ictalurus punctatus) in Southeastern United States. BACKGROUND: Aquaculture systems may contribute to the emergence and persistence of antimicrobial-resistant (AMR) bacteria, posing risks to animal, environmental, and human health. This study characterized the phenotypic and genotypic antimicrobial resistance profiles of Enterococcus spp. isolated from the skin microbiota of 125 channel catfish (Ictalurus punctatus) harvested from two earthen ponds in Alabama, USA. METHODS: Skin swabs from the body of channel catfish were enriched in Enterococcosel broth and cultured on Enterococcosel agar at 28 °C for 24 h. Isolates were confirmed using Biolog Gen III and VITEK(®)2, and antimicrobial susceptibility was determined using the Kirby-Bauer disk diffusion method. Thirty-five randomly sampled isolates underwent whole-genome sequencing for genotypic characterization. RESULTS: 36% of isolates exhibited multidrug resistance (resistance to ≥ 3 antimicrobial classes), with the highest resistance rates observed for ampicillin (44.8%), rifampicin (42.4%), and tetracycline (38.4%). The most prevalent resistance genes were aac(6')-Iid (65.7%), aac(6')-Ii (22.9%), efmA, and msr(C) (20.0% each). Plasmid replicons rep1 and repUS15 frequently co-occurred with resistance genes. Biofilm-associated genes, including efaA, fsrA, fsrB, sprE, ebpABC, ace, and scm, were commonly detected. Multivariate analyses (PERMANOVA, PCA) revealed no significant species-level differences in resistance burden or biofilm gene carriage, indicating similar resistance and virulence gene carriage across species in this dataset. CONCLUSIONS: The skin microbiota of pond-raised catfish harbors antimicrobial-resistant Enterococcus spp. with mobile resistance elements and biofilm-associated virulence factors, suggesting a potential role in AMR persistence within aquaculture settings. These findings support the need for targeted AMR surveillance in fish-associated microbiota as part of integrated One Health strategies.202540760424
1812170.9958Pathogen Detection and Resistome Analysis in Healthy Shelter Dogs Using Whole Metagenome Sequencing. According to the Humane Society, 25 to 40 percent of pet dogs in the United States are adopted from animal shelters. Shelter dogs can harbor bacterial, viral, fungal, and protozoal pathogens, posing risks to canine and human health. These bacterial pathogens may also carry antibiotic resistance genes (ARGs), serving as a reservoir for antimicrobial resistance (AMR) transmission. This study aimed to utilize whole metagenome sequencing (WMS) to screen for microbial pathogens and assess the resistome in healthy shelter dogs. Fecal samples from 58 healthy shelter dogs across 10 shelters in Kentucky, Tennessee, and Virginia were analyzed using WMS. Genomic DNA was extracted, and bioinformatics analyses were performed to identify pathogens and ARGs. The WMS detected 53 potentially zoonotic or known pathogens including thirty-eight bacterial species, two protozoa, five yeast species, one nematode, four molds, and three viruses. A total of 4560 ARGs signatures representing 182 unique genes across 14 antibiotic classes were detected. Tetracycline resistance genes were most abundant (49%), while β-lactam resistance genes showed the highest diversity with 75 unique ARGs. ARGs were predominantly detected in commensal bacteria; however, nearly half (18/38, 47.4%) of known bacterial pathogens detected in this study carried ARGs for resistance to one or more antibiotic classes. This study provides evidence that healthy shelter dogs carry a diverse range of zoonotic and antibiotic-resistant pathogens, posing a transmission risk through fecal shedding. These findings highlight the value of WMS for pathogen detection and AMR surveillance, informing therapeutic and prophylactic strategies to mitigate the transmission of pathogens among shelter dog populations and the risk associated with zoonoses.202539860994
3490180.9958Monitoring and evaluation of antibiotic resistance genes in three rivers in northeast China. Antibiotic resistance genes (ARGs) have become an important public health problem. In this study, we used metagenomic sequencing to analyze the composition of ARGs in selected original habitats of northeast China, comprising three different rivers and riverbank soils of the Heilongjiang River, Tumen River, and Yalu River. Twenty types of ARG were detected in the water samples. The major ARGs were multidrug resistance genes, at approximately 0.5 copies/16S rRNA, accounting for 57.5% of the total ARG abundance. The abundance of multidrug, bacitracin, beta-lactam, macrolide-lincosamide-streptogramin, sulfonamide, fosmidomycin, and polymyxin resistance genes covered 96.9% of the total ARG abundance. No significant ecological boundary of ARG diversity was observed. The compositions of the resistance genes in the three rivers were very similar to each other, and 92.1% of ARG subtypes were shared by all water samples. Except for vancomycin resistance genes, almost all ARGs in riverbank soils were detected in the river water. About 31.05% ARGs were carried by Pseudomonas. Opportunistic pathogenic bacteria carrying resistance genes were mainly related to diarrhea and respiratory infections. Multidrug and beta-lactam resistance genes correlated positively with mobile genetic elements (MGEs), indicating a potential risk of diffusion. The composition of ARGs in three different rivers was similar, indicating that climate plays an important role in ARG occurrence. ARG subtypes in river water were almost completely the same as those in riverbank soil. ARGs had no significant geographical distribution characteristics. Many ARGs were carried by human pathogenic bacteria related to diarrhea and respiratory infections, such as Pseudomonas aeruginosa and Aeromonas caviae. In general, our results provide a valuable dataset of river water ARG distribution in northeast China. The related ecological and geographical distribution characteristics should be further explored.202235122641
2723190.9958Hospital air: A potential route for transmission of infections caused by β-lactam-resistant bacteria. BACKGROUND: The emergence of bacterial resistance to β-lactam antibiotics seriously challenges the treatment of various nosocomial infections. This study was designed to investigate the presence of β-lactam-resistant bacteria (BLRB) in hospital air. METHODS: A total of 64 air samples were collected in 4 hospital wards. Detection of airborne bacteria was carried out using culture plates with and without β-lactams. BLRB isolates were screened for the presence of 5 common β-lactamase-encoding genes. Sequence analysis of predominant BLRB was also performed. RESULTS: The prevalence of BLRB ranged between 3% and 34%. Oxacillin-resistant bacteria had the highest prevalence, followed by ceftazidime- and cefazolin-resistant bacteria. The frequency of β-lactamase-encoding genes in isolated BLRB ranged between 0% and 47%, with the highest and lowest detection for OXA-23 and CTX-m-32, respectively. MecA had a relatively high frequency in surgery wards and operating theaters, whereas the frequency of blaTEM was higher in intensive care units and internal medicine wards. OXA-51 was detected in 4 wards. Acinetobacter spp, Acinetobacter baumannii, and Staphylococcus spp were the most predominant BLRB. CONCLUSIONS: The results revealed that hospital air is a potential route of transmission of BLRB, such as Acinetobacter and Staphylococcus, 2 important causative agents of nosocomial infections. Therefore, improvement of control measures against the spreading of airborne bacteria in hospital environments is warranted.201627021512