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260600.9842Pathogenic multiple antimicrobial resistant Escherichia coli serotypes in recreational waters of Mumbai, India: a potential public health risk. Globally, coastal waters have emerged into a pool of antibiotic resistance genes and multiple antibiotic resistant microorganisms, and pathogenicity of these resistant microorganisms in terms of serotypes and virulence genes has made the environment vulnerable. The current study underscores the presence of multiple antibiotic resistant pathogenic serotypes and pathotypes of Escherichia coli, the predominant faecal indicator bacteria (FIB), in surface water and sediment samples of famous recreational beaches (Juhu, Versova, Mahim, Dadar, and Girgaon) of Mumbai. Out of 65 faecal coliforms (FC) randomly selected, 38 isolates were biochemically characterized, serotyped (for 'O' antigen), antibiogram-phenotyped (for 22 antimicrobial agents), and genotyped by polymerase chain reaction (for virulence factors). These isolates belonged to 16 different serotypes (UT, O141, O2, O119, O120, O9, O35, O126, O91, O128, O87, O86, R, O101, O118, and O15) out of which UT (18.4%), O141 (15.7%), and O2 (13.1%) were predominant, indicating its remarkable diversity. Furthermore, the generated antibiogram profile revealed that 95% of these isolates were multiple antibiotic resistant. More than 60% of aminoglycoside-sensitive E. coli isolates exhibited resistance to penicillin, extended penicillin, quinolone, and cephalosporin classes of antibiotic while resistance to other antibiotics was comparatively less. Antibiotic resistance (AR) indexing indicated that these isolates may have rooted from a high-risk source of contamination. Preliminary findings revealed the presence of enterotoxin-encoding genes (stx1 and stx2 specific for enterohaemorrhagic E. coli and Shiga toxin-producing E. coli, heat-stable toxin enterotoxin specific for enterotoxigenic E. coli) in pathogenic serotypes. Thus, government authorities and environmental planners should create public awareness and adopt effective measures for coastal management to prevent serious health risks associated with these contaminated coastal waters.201728316051
252210.9837Identification and specificity validation of unique and antimicrobial resistance genes to trace suspected pathogenic AMR bacteria and to monitor the development of AMR in non-AMR strains in the environment and clinical settings. The detection of developing antimicrobial resistance (AMR) has become a global issue. The detection of developing antimicrobial resistance has become a global issue. The growing number of AMR bacteria poses a new threat to public health. Therefore, a less laborious and quick confirmatory test becomes important for further investigations into developing AMR in the environment and in clinical settings. This study aims to present a comprehensive analysis and validation of unique and antimicrobial-resistant strains from the WHO priority list of antimicrobial-resistant bacteria and previously reported AMR strains such as Acinetobacter baumannii, Aeromonas spp., Anaeromonas frigoriresistens, Anaeromonas gelatinfytica, Bacillus spp., Campylobacter jejuni subsp. jejuni, Enterococcus faecalis, Escherichia coli, Haemophilus influenzae, Helicobacter pylori, Klebsiella pneumonia subsp. pneumoniae, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serovar Typhimurium, Thermanaeromonas toyohensis, and Vibrio proteolyticus. Using in-house designed gene-specific primers, 18 different antibiotic resistance genes (algJ, alpB, AQU-1, CEPH-A3, ciaB, CMY-1-MOX-7, CMY-1-MOX-9, CMY-1/MOX, cphA2, cphA5, cphA7, ebpA, ECP_4655, fliC, OXA-51, RfbU, ThiU2, and tolB) from 46 strains were selected and validated. Hence, this study provides insight into the identification of strain-specific, unique antimicrobial resistance genes. Targeted amplification and verification using selected unique marker genes have been reported. Thus, the present detection and validation use a robust method for the entire experiment. Results also highlight the presence of another set of 18 antibiotic-resistant and unique genes (Aqu1, cphA2, cphA3, cphA5, cphA7, cmy1/mox7, cmy1/mox9, asaI, ascV, asoB, oxa-12, acr-2, pepA, uo65, pliI, dr0274, tapY2, and cpeT). Of these sets of genes, 15 were found to be suitable for the detection of pathogenic strains belonging to the genera Aeromonas, Pseudomonas, Helicobacter, Campylobacter, Enterococcus, Klebsiella, Acinetobacter, Salmonella, Haemophilus, and Bacillus. Thus, we have detected and verified sets of unique and antimicrobial resistance genes in bacteria on the WHO Priority List and from published reports on AMR bacteria. This study offers advantages for confirming antimicrobial resistance in all suspected AMR bacteria and monitoring the development of AMR in non-AMR bacteria, in the environment, and in clinical settings.202338058762
278220.9836Urban dust fecal pollution in Mexico City: antibiotic resistance and virulence factors of Escherichia coli. Fecal pollution of settled dust samples from indoor and outdoor urban environments, was measured and characterized by the presence of fecal coliforms (FC), and by the characterization of Escherichia coli virulence genes, adherence and antibiotic resistance traits as markers. There were more FC indoors than outdoors (mean values 1089 and 435MPN/g). Among indoor samples, there were more FC in houses with carpets and/or pets. Using a PCR-based assay for six enteropathogenicity genes (belonging to the EAEC, EHEC and EPEC pathotypes) on randomly selected E. coli isolates, there was no significant difference between isolates from indoors and outdoors (60% and 72% positive to at least one gene). The serotypes commonly associated with pathogenic strains, such as O86 and O28, were found in the indoor isolates; whereas O4, O66 and O9 were found amongst outdoor isolates. However, there were significantly more outdoor isolates resistant to at least one antibiotic (73% vs. 45% from indoors) among the strains positive for virulence factors, and outdoor isolates were more commonly multiresistant. There was no relationship between the presence of virulence genes and resistance traits. These results indicate that outdoor fecal bacteria were more likely from human sources, and those found indoors were related to pets and maintained in carpets. This study illustrates the risk posed by fecal bacteria from human sources, usually bearing virulence and resistance traits. Furthermore, the high prevalence of strains carrying genes associated to EAEC or EHEC pathotypes, in both indoor and outdoor environments, adds to the health risk.200616762593
134030.9835Prevalence, Virulence, and Antimicrobial Resistance of Campylobacter spp. in Raw Milk, Beef, and Pork Meat in Northern Poland. The purpose of this study was to determine whether raw milk, unpasteurized dairy products, pork, and beef available for sale in the Kujawsko-Pomorskie and Wielkopolska regions in Poland are contaminated with Campylobacter spp. bacteria and may be a potential source of infection. For isolated strains, antibiotic susceptibility and the presence of genes responsible for virulence were examined. Material for research included 1058 food samples collected between 2014 and 2018 with 454 samples of raw milk and unpasteurized dairy products (milk from vending machines, milk from owners of dairy cows, cheese, milk cream) and 604 samples of raw meat (pork, beef). The results indicated that 9.3% of the samples were positive for Campylobacter spp., and Campylobacter jejuni was predominant in this study. Campylobacter bacteria was not found in milk collected from vending machines, as well as cheese and milk cream samples. Campylobacter was noted in 12.7% of beef samples, 11.8% of raw milk purchased from individual suppliers, and 10.9% of pork samples. Resistance to erythromycin (2.0%), azithromycin (3.1%), gentamicin (4.1%), tetracycline (65.3%), and ciprofloxacin (71.4%) was determined using the disc diffusion method. Furthermore, the prevalence of racR, sodB, csrA, virB11, cdtB, iam, and wlaN genes were examined using the PCR method. The sodB, csrA, and cdtB genes exhibited the highest detection rate, but none of the genes were identified in 100% of the isolates. Statistically significant differences between the presence of virulence marker genes, including for iam, racR, and csrA markers, were noted among different sources of the isolates. Differences in the distribution of iam, wlaN, and virB11 were also shown between C. jejuni and C. coli strains. As a result of the analysis, it has been concluded that unpasteurized milk, beef, and pork could be a sources of Campylobacter pathogens. Moreover, this study revealed virulent properties of Campylobacter isolated from such food products and high resistance rates to fluoroquinolones, which may represent difficulties in campylobacteriosis treatment.201931533265
278340.9833Prevalence of β-lactamase genes in domestic washing machines and dishwashers and the impact of laundering processes on antibiotic-resistant bacteria. AIMS: To investigate the prevalence of β-lactamase genes in domestic washing machines and dishwashers, and the decontamination efficacy of laundering. METHODS AND RESULTS: For the first investigation, swab samples from washing machines (n = 29) and dishwashers (n = 24) were analysed by real-time quantitative PCR to detect genes encoding β-lactamases. To test the impact of laundering on resistant bacteria, cotton test swatches were artificially contaminated with susceptible and resistant strains of Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus within a second investigation. They were washed in a domestic washing machine with or without activated oxygen bleach (AOB)-containing detergent at 20-50°C. β-Lactamase genes (most commonly of the AmpC- and OXA-type) were detected in 79% of the washing machines and in 96% of the dishwashers and Pseudomonadaceae dominated the microbiota. The level of bacterial reduction after laundering was ≥80% for all Ps. aeruginosa and Kl. pneumoniae strains, while it was only 37-61% for the methicillin-resistant Staph. aureus outbreak strain. In general, the reduction was tendentially higher for susceptible bacteria than for the resistant outbreak strains, especially for Staph. aureus. CONCLUSIONS: β-Lactamase genes seem to be frequently present in domestic appliances and may pose a potential risk for cross-contamination and horizontal transfer of genes encoding resistance against clinically important β-lactams. In general, higher temperatures and the use of AOB can improve the reduction of antibiotic-resistant bacteria, including Staph. aureus which appears to be less susceptible to the decontamination effect of laundering. SIGNIFICANCE AND IMPACT OF THIS STUDY: Data on the presence of antibiotic-resistant bacteria in the domestic environment are limited. This study suggests that β-lactamase genes in washing machines and dishwashers are frequent, and that antibiotic-resistant strains are generally more resistant to the used washing conditions.201728845592
258250.9833The Household Resistome: Frequency of β-Lactamases, Class 1 Integrons, and Antibiotic-Resistant Bacteria in the Domestic Environment and Their Reduction during Automated Dishwashing and Laundering. Households provide a habitat for bacteria originating from humans, animals, foods, contaminated clothes, or other sources. Thus, bacteria carrying antibiotic resistance genes (ARGs) may be introduced via household members, animals, or the water supply from external habitats into private households and vice versa. Since data on antibiotic resistance (ABR) in the domestic environment are limited, this study aimed to determine the abundance of β-lactamase, mobile colistin resistance, and class 1 integron genes and the correlation of their presence and to characterize phenotypically resistant strains in 54 private households in Germany. Additionally, the persistence of antibiotic-resistant bacteria during automated dishwashing compared to that during laundering was assessed. Shower drains, washing machines, and dishwashers were sampled and analyzed using quantitative real-time PCR. Resistant strains were isolated, followed by identification and antibiotic susceptibility testing using a Vitek 2 system. The results showed a significantly higher relative ARG abundance of 0.2367 ARG copies/16S rRNA gene copies in shower drains than in dishwashers (0.1329 ARG copies/16S rRNA gene copies) and washing machines (0.0006 ARG copies/16S rRNA gene copies). bla(CMY-2), bla(ACT/MIR), and bla(OXA-48) were the most prevalent ARG, and intI1 occurred in 96.3% of the households, while no mcr genes were detected. Several β-lactamase genes co-occurred, and the resistance of bacterial isolates correlated positively with genotypic resistance, with carbapenemase genes dominating across isolates. Antibiotic-resistant bacteria were significantly reduced during automated dishwashing as well as laundering tests and did not differ from susceptible strains. Overall, the domestic environment may represent a potential reservoir of β-lactamase genes and β-lactam-resistant bacteria, with shower drains being the dominant source of ABR.IMPORTANCE The abundance of antibiotic-resistant bacteria and ARGs is steadily increasing and has been comprehensively analyzed in natural environments, animals, foods, and wastewater treatment plants. In this respect, β-lactams and colistin are of particular interest due to the emergence of multidrug-resistant Gram-negative bacteria. Despite the connection of private households to these environments, only a few studies have focused on the domestic environment so far. Therefore, the present study further investigated the occurrence of ARGs and antibiotic-resistant bacteria in shower drains, washing machines, and dishwashers. The analysis of the domestic environment as a potential reservoir of resistant bacteria is crucial to determine whether households contribute to the spread of ABR or may be a habitat where resistant bacteria from the natural environment, humans, food, or water are selected due to the use of detergents, antimicrobial products, and antibiotics. Furthermore, ABR could limit the options for the treatment of infections arising in the domestic environment.202032978137
240860.9830Prevalence and Detection of qac Genes from Disinfectant-Resistant Staphylococcus aureus Isolated from Salon Tools in Ishaka Town, Bushenyi District of Uganda. Bacterial infections are on a rise with causal-resistant strains increasing the economic burden to both patients and healthcare providers. Salons are recently reported as one of the sources for transmission of such resistant bacterial strains. The current study aimed at the identification of the prevalent bacteria and characterization of quaternary ammonium compound (qac) genes from disinfectant-resistant S. aureus isolated from salon tools in Ishaka town, Bushenyi District of Uganda. A total of 125 swabs were collected from different salon tools (combs, brushes, scissors, clippers, and shaving machines), and prevalent bacteria were isolated using standard microbiological methods. Identification of isolated bacteria was done using standard phenotypic methods including analytical profile index (API). Susceptibility patterns of the isolated bacteria to disinfectant were determined using the agar well diffusion method. Quaternary ammonium compound (qac) genes (qacA/B and qacC) associated with disinfectant resistances were detected from disinfectant-resistant S. aureus using multiplex polymerase chain reaction (PCR) and Sanger sequencing methods. Of the 125 swab samples collected from salons, 78 (62.4%) were contaminated with different bacteria species. Among the salon tools, clippers had the highest contamination of 20 (80.0%), while shaving machines had the lowest contamination of 11 (44.0%). The most prevalent bacteria identified were Staphylococcus epidermidis (28.1%) followed by S. aureus (26.5%). Of all the disinfectants tested, the highest resistance was shown with sodium hypochlorite 1%. Out of the eight (8) disinfectant-resistant S. aureus analysed for qac genes, 2 (25%) isolates (STP6 and STP9) were found to be qacA/B positive, while 2 (25%) isolates (STP8 and STP9) were found to be qacC gene positive. This study has shown that bacterial contamination of salon tools is common, coupled with resistance to disinfectants with sodium hypochlorite resistance being more common. Furthermore, observed resistance was attributed to the presence of qac genes among S. aureus isolates. A search for qac genes for disinfectant resistance from other bacteria species is recommended.202032849931
121170.9828Molecular characterization of multidrug-resistant Escherichia coli of the phylogroups A and C in dairy calves with meningitis and septicemia. Escherichia coli is an important cause of septicemia (SEPEC) and neonatal meningitis (NMEC) in dairy calves. However, the diversity of virulence profiles, phylogroups, antimicrobial resistance patterns, carriage of integron structures, and fluoroquinolone (FQ) resistance mechanisms have not been fully investigated. Also, there is a paucity of knowledge about the virulence profiles and frequency of potential SEPEC in feces from calves with or without diarrhea. This study aimed to characterize the virulence potential, phylogroups, antimicrobial susceptibility, integron content, and FQ-resistance mechanisms in Escherichia coli isolated from calves with meningitis and septicemia. Additionally, the virulence genes (VGs) and profiles of E. coli isolated from diarrheic and non-diarrheic calves were compared between them and together with NMEC and SEPEC in order to identify shared profiles. Tissue and fluid samples from eight dairy calves with septicemia, four of which had concurrent meningitis, were processed for bacteriology and histopathology. Typing of VGs was assessed in 166 isolates from diverse samples of each calf. Selected isolates were evaluated for antimicrobial susceptibility by the disk diffusion test. Phylogroups, integron gene cassettes cartography, and FQ-resistance determinants were analyzed by PCR, sequencing, and bioinformatic tools. Furthermore, 109 fecal samples and 700 fecal isolates from dairy calves with or without diarrhea were evaluated to detect 19 VGs by uniplex PCR. Highly diverse VG profiles were characterized among NMEC and SEPEC isolates, but iucD was the predominant virulence marker. Histologic lesions in all calves supported their pathogenicity. Selected isolates mainly belonged to phylogroups A and C and showed multidrug resistance. Classic (dfrA17 and arr3-dfrA27) and complex (dfrA17-aadA5::ISCR1::bla(CTX-M-2)) class 1 integrons were identified. Target-site mutations in GyrA (S83L and D87N) and ParC (S80I) encoding genes were associated with FQ resistance. The VGs detected more frequently in fecal samples included f17G (50%), papC (30%), iucD (20%), clpG (19%), eae (16%), and afaE-8 (13%). Fecal isolates displaying the profiles of f17 or potential SEPEC were found in 25% of calves with and without diarrhea. The frequency of E. coli VGs and profiles did not differ between both groups (p > 0.05) and were identical or similar to those found in NMEC and SEPEC. Overall, multidrug-resistant E. coli isolates with diverse VG profiles and belonging to phylogroups A and C can be implicated in natural cases of meningitis and septicemia. Their resistance phenotypes can be partially explained by class 1 integron gene cassettes and target-site mutations in gyrA and parC. These results highlight the value of antimicrobial resistance surveillance in pathogenic bacteria isolated from food-producing animals. Besides, calves frequently shed potential SEPEC in their feces as commensals ("Trojan horse"). Thus, these bacteria may be disseminated in the farm environment, causing septicemia and meningitis under predisposing factors.202234982979
248780.9828Clinical cases, drug resistance, and virulence genes profiling in Uropathogenic Escherichia coli. Uropathogenic Escherichia coli (UPEC) as the most important bacterial agent of urinary tract infections (UTIs) encompasses a wide treasure of virulence genes and factors. In due to this default, the aim of this research was to detect and identify some important virulence genes including cnf1, upaH, hlyA, ibeA, and cdtB in isolated UPEC pathotypes. In this research, clinical samples of urine were collected in Shahr-e-Qods, Tehran, Iran. The UPEC pathotypes were confirmed by standard biochemical tests. The DNAs of isolated bacteria were extracted. The genes of cnf1, upaH, hlyA, ibeA, and cdtB were run for multiplex PCR and gel electrophoresis. Furthermore, the antibiogram was done for the isolated UPEC strains by 11 common antibiotics. In accordance with the results, the virulence genes of cnf1, upaH, hlyA, ibeA, and cdtB were respectively recognized in 100%, 51.2%, 38.4%, 9.3%, and 0% of isolated UPEC pathotypes. In consequence, the final virulence gene profiling of the isolated UPEC strains was patterned as cnf1, cnf1-upaH, cnf1-upaH-hlyA, and cnf1-upaH-hlyA-ibeA. The chi-square tests showed no significant correlations between virulence gene profile and UTIs, between virulence gene profile and antibiotic resistance, and between virulence genes and different types of UTIs. The cnf1 virulence gene contributes in the occurrence of all types of UTIs. In contrast to cnf1, the cdtB gene was absent in the isolated UPEC strains in this investigation. The most ineffective antibiotics were recognized as Penicillin, Tetracycline, and Nalidixic acid, respectively, while Streptomycin, Chloramphenicol, and Ciprofloxacin are the best options for UTIs treatment.202031950434
236890.9828Smelly shark, smelly ray: what is infecting you? AIMS: Although elasmobranchs are consumed worldwide, bacteriological assessments for this group are still sorely lacking. In this context, this study assessed bacteria of sharks and rays from one of the most important landing ports along the Rio de Janeiro coast. METHODS AND RESULTS: Bacteria were isolated from the cloacal swabs of the sampled elasmobranchs. They were cultured, and Vibrio, Aeromonas, and Enterobacterales were isolated and identified. The isolated bacteria were then biochemically identified and antimicrobial susceptibility assays were performed. Antigenic characterizations were performed for Salmonella spp. and Polymerase Chain Reaction (PCR) assays were performed to identify Escherichia coli pathotypes. Several bacteria of interest in the One Health context were detected. The most prevalent Enterobacterales were Morganella morganii and Citrobacter freundii, while Vibrio harveyi and Vibrio fluvialis were the most prevalent among Vibrio spp. and Aeromonas allosacharophila and Aeromonas veronii bv. veronii were the most frequent among Aeromonas spp. Several bacteria also displayed antimicrobial resistance, indicative of Public Health concerns. A total of 10% of Vibrio strains were resistant to trimethoprim-sulfamethoxazole and 40% displayed intermediate resistance to cefoxitin. Salmonella enterica strains displayed intermediate resistance to ciprofloxacin, nalidixic acid and streptomycin. All V. cholerae strains were identified as non-O1/non-O139. The detected E. coli strains did not exhibit pathogenicity genes. This is the first study to perform serology assessments for S. enterica subsp. enterica isolated from elasmobranchs, identifying the zoonotic Typhimurium serovar. Salmonella serology evaluations are, therefore, paramount to identify the importance of elasmobranchs in the epidemiological salmonellosis chain. CONCLUSIONS: The detection of several pathogenic and antibiotic-resistant bacteria may pose significant Public Health risks in Brazil, due to high elasmobranch consumption rates, indicating the urgent need for further bacteriological assessments in this group.202438486350
2099100.9828Antibiotic-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
1354110.9827The prevalence, antibiotic resistance and multilocus sequence typing of colistin-resistant bacteria isolated from Penaeus vannamei farms in earthen ponds and HDPE film-lined ponds in China. The aquaculture environment, especially the culture ponds and aquaculture products, is considered to be an important reservoir of colistin resistance genes. However, systematic investigations of colistin resistance in Penaeus vannamei farming in different culture modes are scarce. In this study, a total of 93 non-duplicated samples were collected from P. vannamei farms in five cities in China from 2019 to 2021. The prevalence, antibiotic resistance and multilocus sequence typing (MLST) of colistin-resistant bacteria were measured and analysed. The results showed that among the 1601 isolates in P. vannamei and its environmental samples, the pollution of colistin-resistant bacteria was serious (the overall prevalence was 37.3% and 28.8%, respectively), regardless of the earthen pond or high-density polyethylene (HDPE) film-lined pond. Among 533 isolates, the prevalence of mobile colistin resistance (mcr) genes, mcr-1, was the highest (60%, 320/533), followed by mcr-4 (1.5%, 8/533), mcr-8 (0.9%, 5/533), mcr-10 (0.6%, 3/533) and mcr-7 (0.4%, 2/533). The prevalence of mcr-1 in earthen ponds was significantly higher than that in HDPE film-lined ponds (67.5% vs. 49.1%, p < .001). The dominant strain carrying mcr-1 was Bacillus spp. (54.1%, 173/320), followed by Enterobacter spp. (8.1%, 26/320), Staphylococcus spp. (6.3%, 20/320) and Aeromonas spp. (5.3%, 17/320). The antibiotic resistance profiles of 173 Bacillus spp. varied among different sampling locations and culture types. These isolates were highly resistant to cefepime, ceftriaxone, trimethoprim-sulfamethoxazole and ceftiofur (>45%), and multidrug-resistant isolates were common (62.4%, 108/173). Sequence type (ST) 26 (37/66, 56%) was found to be the most prevalent ST in mcr-1-positive Bacillus cereus isolated from the aquaculture environment. In summary, our study pointed out that it is necessary to continuously monitor antibiotic usage and its residues regardless of the pond types, especially with regard to critical drugs such as colistin.202235841601
2725120.9827Hygiene 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
5607130.9826Phenotypic and genotypic characterization of antimicrobial resistance and virulence profiles of Salmonella enterica serotypes isolated from necropsied horses in Kentucky. Salmonella is a foodborne pathogen that poses a significant threat to global public health. It affects several animal species, including horses. Salmonella infections in horses can be either asymptomatic or cause severe clinical illness. Infections caused by Salmonella are presently controlled with antibiotics. Due to the formation of biofilms and the emergence of antimicrobial resistance, the treatment has become more complicated. Our study focused on investigating the prevalence of Salmonella enterica in necropsied horses, assessing the capability for biofilm formation, and motility, determining the phenotypic and genotypic profiles of antibiotic resistance, and detecting virulence genes. A total of 2,182 necropsied horses were tested for the presence of Salmonella. Intestinal samples were enriched in selenite broth and cultured on hektoen and eosin methylene blue agar plates, whereas other samples were directly cultured on aforementioned plates. Confirmation of the serotypes was performed according to the Kauffmann-White-Le Minor Scheme followed by biofilm formation screening using crystal violet assay. The resistance profile of the isolates was determined by broth microdilution assay using the Sensititre️ Vet (Equine EQUIN2F). The genotypic antimicrobial resistance (AMR) and virulence profiles were detected using polymerase chain reaction (PCR). The overall prevalence of Salmonella was 1.19% (26/2182), with 11 different serotypes identified. Salmonella Typhimurium was the most prevalent serotype with 19.2% prevalence. All of the isolates were identified as biofilm producers and motile. Virulence genes related to invasion (invA, hilA, mgtC, and spiA), biofilm formation (csgA and csgB), and motility (filA, motA, flgG, figG, flgH, fimC, fimD, and fimH) of Salmonella were detected among 100% of the isolates. An overall 11.4% of the isolates were identified as multidrug-resistant (MDR), with resistance to gentamicin, amikacin, ampicillin, ceftazidime, ceftiofur, chloramphenicol, and trimethoprim/sulfamethoxazole. We found that beta-lactamase-producing genes bla(TEM), bla(CTXM), and bla(SHV2) were identified in 11.5% of the isolates, while only 3.8% carried the bla(OXA-9) gene. The presence of MDR pathogenic Salmonella in horses is alarming for human and animal health, especially when they have a high affinity for forming biofilm. Our study found horses as potential sources of pathogenic Salmonella transmission to humans. Thus, it is important to perform continuous monitoring and surveillance studies to track the source of infection and develop preventive measures. IMPORTANCE: This study focuses on understanding how Salmonella, specifically isolated from horses, can resist antibiotics and cause disease. Salmonella is a well-known foodborne pathogen that can pose risks not only to animals but also to humans. By studying the bacteria from necropsied horses, the research aims to uncover how certain Salmonella strains develop resistance to antibiotics and which genetic factors make them more dangerous. In addition to antibiotic resistance, the research explores the biofilm-forming ability of these strains, which enhances their survival in harsh environments. The study also investigates their motility, a factor that contributes to the spread of infection. The findings can improve treatment strategies for horses and help prevent the transmission of resistant bacteria to other animals as well as humans. Ultimately, the research could contribute to better management of antibiotic resistance in both veterinary and public health contexts, helping to safeguard animal welfare and public health.202539846771
2584140.9825Resistance 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
2706150.9824Prevalence and antimicrobial resistance profile of bacterial foodborne pathogens in Nile tilapia fish (Oreochromis niloticus) at points of retail sale in Nairobi, Kenya. Proteus spp., Staphylococcus spp., Pseudeomonas spp., and pathogenic Vibrios are among the major foodborne pathogens associated with the consumption of contaminated fish. The increasing occurrence of antimicrobial resistance in these pathogens is a serious public health concern globally and therefore continuous monitoring of antimicrobial resistance of these bacteria along the food chain is crucial for for control of foodborne illnesses. The aim of this study was to assess the prevalence, antimicrobial resistance patterns, antibiotic resistance genes, and genetic diversity of bacterial foodborne pathogens recovered from fresh Nile tilapia (Oreochromis niloticus) obtained from retail markets in Nairobi, Kenya. A total of 68 O. niloticus fish with an average weight of 300.12 ± 25.66 g and body length of 23.00 ± 0.82 cm were randomly sampled from retail markets and tested for the presence of Proteus, Staphylococcus aureus, Pseudomonas aeruginosa, Vibrio cholerae, and Vibrio parahaemolyticus. Standard culture-based microbiological and Kirby-Bauer agar disk diffusion methods were used to isolate and determine the antimicrobial resistance patterns of the isolates to 11 selected antibiotics. Statistical analysis was performed using Minitab v17.1, with p < 0.05 considered significant. The genetic diversity of the multidrug-resistant (MDR) and extensively drug-resistant (XDR) bacteria was determined using 16S rRNA sequencing and phylogenetic analysis, and polymerase chain reaction (PCR) was used for detection of antibiotic resistance genes in MDR bacterial isolates. High levels of bacterial contamination were detected in fresh O. niloticus fish (44/68, 64.71%). The most prevalent bacteria were Proteus spp. (44.12%), with the rest of the bacterial species registering a prevalence of 10.29%, 4.41%, 2.94%, and 2.94% (for S. aureus, P. aeruginosa, V. cholerae, and V. parahaemolyticus, respectively). Antimicrobial resistance was detected in all the bacteria species and all the isolates were resistant to at least one antibiotic except cefepime (30 µg). Additionally, 86.36% of the isolates exhibited multidrug resistance, with higher multiple antibiotic resistance indices (MAR index >0.3) indicating that fresh O. niloticus fish were highly contaminated with MDR bacteria. Results of 16S rRNA sequences, BLASTn analysis, and phylogenetic trees confirmed the identified MDR bacterial isolates as Proteus mirabilis and other Proteus spp., S. aureus, P. aeruginosa, V. cholerae, and V. parahaemolyticus. PCR analysis confirmed the presence of multiple antibiotic resistance genes blaTEM-1, blaCMY-2, tetA, tetC, Sul2, dfrA7, strA, and aadA belonging to β-lactamases, tetracycline, sulfonamide, trimethoprim, and aminoglycosides in all the MDR bacterial isolates. There was strong correlation between antibiotic- resistant genes and phenotypic resistance to antibiotics of MDR bacteria. This study showed high prevalence of multidrug resistance among foodborne bacterial isolates from fresh O. niloticus fish obtained from retail markets. From this study, we conclude that fresh O. niloticus fish are a potential source of MDR bacteria, which could be a major risk to public health as a consequence of their dissemination along the human food chain. These results highlight the prevalence of antimicrobial-resistant foodborne pathogens in fish purchased from retail markets and underscore the risk associated with improper handling of fish.202339816642
2876160.9824High prevalence of multiple-antibiotic-resistant (MAR) Escherichia coli in river bed sediments of the Apies River, South Africa. This study aimed at investigating the presence of antibiotic-resistant Escherichia coli in river bed sediments of the Apies River, Gauteng, South Africa, in order to better inform health management decisions designed to protect users of the river. Overall, 180 water and sediment samples were collected at 10 sites along the Apies River from January to February 2014. E. coli was enumerated using the Colilert® 18/Quanti-Tray® 2000 (IDEXX). Isolates were purified by streaking on eosin methylene blue agar followed by the indole test. Pure E. coli isolates were tested for resistance to nine antibiotics by the Kirby-Bauer disc diffusion method. Over 98% of the isolates were resistant to at least one of the antibiotics tested. The highest resistance was observed against nitrofurantoin (sediments) and ampicillin (water). Over 80% of all resistant isolates showed multiple antibiotic resistance (resistance to ≥3 antibiotics). The abundance of E. coli in the sediments not only adds to the evidence that sediments are a reservoir for bacteria and possibly other pathogens including antibiotic-resistant bacteria but also suggests that antibiotic-resistant genes could be transferred to pathogens due to the high prevalence of multiple-antibiotic-resistant (MAR) strains of E. coli observed in the sediment. Using untreated water from the Apies River following resuspension for drinking and other household purposes could pose serious health risks for users. Our results suggest that river bed sediments could serve as reservoirs for MAR bacteria including pathogens under different climatic conditions and their analysis could provide information of public health concerns.201526419380
1333170.9824Virulence-encoding genes related to extraintestinal pathogenic E. coli and multidrug resistant pattern of strains isolated from neonatal calves with different severity scores of umbilical infections. Umbilical infections in calves comprise a major cause of neonatal mortality and have been related to a variety of microorganisms. E. coli is an opportunistic enteropathogen characterized by a diversity of virulence factors (VF). Nonetheless, the gene profiles that encode VF associated with umbilical infections in calves and their effect on the clinical severity remains unclear. In this scenario, microbial identification (with an emphasis on E. coli), was carried out among 150 neonatal calves (≤30 days of age) with umbilical infections, where the omphalopathies were clinically scored as mild, moderate, or severe. Also, a panel of 16 virulence-encoding genes related to extraintestinal pathogenic E. coli (ExPEC) were investigated, i.e., fimbriae/adhesins (sfa/focDEa, papA, papC, afaBC), toxins (hlyA, sat, cnf1, cdt), siderophores (iroN, irp2, iucD, ireA), invasins (ibeA), and serum resistance (ompT, traT, kpsMT II). Bacteria and yeasts isolates were identified using mass spectrometry. Bacteria, yeasts, and fungi were isolated in 94.7% (142/150) of neonatal calves sampled. E. coli was the agent most frequently isolated (59/150 = 39.3%), in pure culture (27/59 = 45.8%) and combined infections (32/59 = 54.2%), although a great variety (n = 83) of other species of microorganisms were identified. Clinical severity scores of 1, 2, and 3 were observed in 32.2% (19/59), 23.7% (14/59), and 44.1% (26/59) of E. coli infections, respectively. The ExPEC genes detected were related to serum resistance (traT, 42/59 = 72.2%; ompT, 35/59 = 59.3%, kpsMTII, 10/59 = 17%), invasins (ibeA, 11/59 = 18.6%), siderophores (iucD, 9/59 = 15.3%; iroN, 8/59 = 13.6%), and adhesins/fimbriae (papA, 8/59 = 13.6%; papC, 15/59 = 9.6%). The presence of each virulence gene was not associated with the case's clinical score. Among all isolates, 89.8% (53/59) showed in vitro resistance to sulfamethoxazole/trimethoprim and 59.3% to ampicillin (35/59), while 94.1% (55/59) revealed a multidrug resistant profile. Great complexity of bacteria, yeast, and fungi species was identified, reinforcing the umbilical infections of neonatal calves as a polymicrobial disorder. The high occurrence of E. coli (39.3%) highlights the role of this pathogen in the etiology of umbilical infections in calves. Furthermore, a panel of ExPEC genes was investigated for the first time among calves that were clinically scored for case severity. The high prevalence of traT and ompT indicates that these serum resistance-related genes could be used as biomarkers for further investigations of ExPEC isolates from umbilical infections. Our results contribute to the etiological investigation, clinical severity scoring, antimicrobial resistance pattern, and virulence-related to ExPEC genes involved in umbilical infections of neonatal calves.202336427660
1345180.9824Toxigenic potential and antimicrobial susceptibility of Bacillus cereus group bacteria isolated from Tunisian foodstuffs. BACKGROUND: Despite the importance of the B. cereus group as major foodborne pathogens that may cause diarrheal and/or emetic syndrome(s), no study in Tunisia has been conducted in order to characterize the pathogenic potential of the B. cereus group. The aim of this study was to assess the sanitary potential risks of 174 B. cereus group strains isolated from different foodstuffs by detecting and profiling virulence genes (hblA, hblB, hblC, hblD, nheA, nheB, nheC, cytK, bceT and ces), testing the isolates cytotoxic activity on Caco-2 cells and antimicrobial susceptibility towards 11 antibiotics. RESULTS: The entertoxin genes detected among B. cereus isolates were, in decreasing order, nheA (98.9%), nheC (97.7%) and nheB (86.8%) versus hblC (54.6%), hblD (54.6%), hblA (29.9%) and hblB (14.9%), respectively encoding for Non-hemolytic enterotoxin (NHE) and Hemolysin BL (HBL). The isolates are multi-toxigenic, harbouring at least one gene of each NHE and HBL complexes associated or not to bceT, cytK-2 and ces genes. Based on the incidence of virulence genes, the strains were separated into 12 toxigenic groups. Isolates positive for cytK (37,9%) harbored the cytK-2 variant. The detection rates of bceT and ces genes were 50.6 and 4%, respectively. When bacteria were incubated in BHI-YE at 30 °C for 18 h and for 5 d, 70.7 and 35% of the strains were shown to be cytotoxic to Caco-2 cells, respectively. The cytotoxicity of B. cereus strains depended on the food source of isolation. The presence of virulence factors is not always consistent with cytotoxicity. However, different combinations of enterotoxin genetic determinants are significantly associated to the cytotoxic potential of the bacteria. All strains were fully sensitive to rifampicin, chloramphenicol, ciprofloxacin, and gentamycin. The majority of the isolates were susceptible to streptomycin, kanamycin, erythromycin, vancomycin and tetracycline but showed resistance to ampicillin and novobiocin. CONCLUSION: Our results contribute data that are primary to facilitate risk assessments in order to prevent food poisoning due to B. cereus group.201931445510
2791190.9823Molecular characterization, antibiotic resistant pattern and biofilm forming potentiality of bacterial community associated with Ompok pabda fish farming in southwestern Bangladesh. Ompok pabda is gaining popularity in the aquaculture industry due to its increasing demand; however research on microbial diversity and antibiotic susceptibility remains limited. The present study was designed to identify the bacterial pathogens commonly found in the pabda farming system with their biofilm forming potential and antibiotic susceptibility. Different bacterial strains were isolated from water, sediments and gut, gill of pabda fish and the isolates were identified based on their morphological traits, biochemical and molecular analysis. Antibiotic susceptibilities, antibiotic resistance gene determination and biofilm formation capabilities were evaluated by disc diffusion method, PCR amplification and Microtiter plate (MTP) assay, respectively. The respective isolates of gill and gut of pabda aquaculture and their environments were: Exiguobacterium spp. (25 %), Enterococcus spp. (20 %), Bacillus spp. (10 %), Acinetobacter spp. (10 %), Enterobacter spp. (10 %), Aeromonas spp. (10 %), Lactococcus spp. (5 %), Klebsiella spp. (5 %) and Kurthia spp. (5 %). Antibiotic resistance frequencies were found to be relatively high, especially for trimethoprim (95 %), sulfafurazole (75 %), ampicillin (60 %), amoxicillin-clavulanic acid (55 %), and cephradine (50 %). 30 % isolates were categorized as DR bacteria followed by 30 % isolates were MDR bacteria and 40 % were classified as XDR bacteria. Moreover, 4 antibiotic resistant genes were detected with sul1 (30 %), dfrA1 (10 %), tetC (40 %), and qnrA (5 %) of isolates. Based on the microtiter plate method, 20 %, 25 %, and 30 % of isolates were found to produce strong, moderate, and weak biofilms, respectively. The findings suggest that biofilm forming bacterial strains found in O. pabda fish farm may be a potential source of numerous antibiotic-resistant bacteria. The study sheds new light on antibiotic resistance genes, which are typically inherited by bacteria and play an important role in developing effective treatments or control strategies.202439047804