Molecular characterization of Vibrio species isolated from dairy and water samples. - Related Documents




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288201.0000Molecular characterization of Vibrio species isolated from dairy and water samples. Vibrio species can cause foodborne infections and lead to serious gastrointestinal illnesses. The purpose of this research was to detect the Vibrio cholerae and Vibrio parahaemolyticus in raw milk, dairy products, and water samples. Also, it investigated the virulence factors, antibiotic resistance and biofilm formation in isolated bacteria. Conventional and molecular approaches were used to identify the isolates in this study. Vibrio species were detected in 5% of the samples. Vibrio cholerae and Vibrio parahaemolyticus were isolated from 1.25 and 1.5%, respectively, of the total samples. Penicillin resistance was detected in all strains of Vibrio cholerae and Vibrio parahaemolyticus, with a MAR index ranging from 0.16 to 0.5. Four isolates were moderate biofilm producer and three of them were MDR. When Vibrio cholerae was screened for virulence genes, ctxAB, hlyA, and tcpA were found in 80, 60, and 80% of isolates, respectively. However, tdh + /trh + associated-virulence genes were found in 33.3% of Vibrio parahaemolyticus isolates.202337717062
271810.9999Virulence-Associated Genes and Antimicrobial Resistance of Aeromonas hydrophila Isolates from Animal, Food, and Human Sources in Brazil. Aeromonads are natural inhabitants of aquatic environments and may be associated with various human or animal diseases. Its pathogenicity is complex and multifactorial and is associated with many virulence factors. In this study, 110 selected Aeromonas hydrophila isolates isolated from food, animals, and human clinical material from 2010 to 2015 were analyzed. Antimicrobial susceptibility testing was performed by the disk diffusion method, and polymerase chain reaction was conducted to investigate the virulence genes hemolysin (hlyA), cytotoxic enterotoxin (act), heat-labile cytotonic enterotoxin (alt), aerolysin (aerA), and DNase-nuclease (exu). At least 92.7% of the isolates had one of the investigated virulence genes. Twenty different virulence profiles among the isolates were recognized, and the five investigated virulence genes were observed in four isolates. Human source isolates showed greater diversity than food and animal sources. Antimicrobial resistance was observed in 46.4% of the isolates, and multidrug resistance was detected in 3.6% of the isolates. Among the 120 isolates, 45% were resistant to cefoxitin; 23.5% to nalidixic acid; 16.6% to tetracycline; 13.7% to cefotaxime and imipenem; 11.8% to ceftazidime; 5.9% to amikacin, gentamicin, and sulfamethoxazole-trimethoprim; and 3.9% to ciprofloxacin and nitrofurantoin. Overall, the findings of our study indicated the presence of virulence genes and that antimicrobial resistance in A. hydrophila isolates in this study is compatible with potentially pathogenic bacteria. This information will allow us to recognize the potential risk through circulating isolates in animal health and public health and the spread through the food chain offering subsidies for appropriate sanitary actions.202032461959
590020.9998Safety profiles of beneficial lactic acid bacteria isolated from dairy systems. This study aimed to assess the safety aspects of 15 lactic acid bacteria (LAB) strains previously isolated from a dairy environment with relation to their beneficial features. LAB strains were assessed using phenotypic methods according to their production of virulence factors at 25 °C and 37 °C, as well as by examining their potential resistance to 15 antibiotics. Polymerase chain reaction (PCR) was also used to identify the presence of 50 genes associated with virulence factors and antibiotic resistance in the strains. None of the strains presented hemolytic activity or the production of gelatinase, lipase, deoxyribonuclease, or the tested biogenic amines. Based on the disk diffusion assay, all strains were resistant to oxacillin and sulfa/trimethoprim. Further, some were resistant to gentamicin (14), clindamycin (11), vancomycin (9), rifampicin (8), erythromycin (5), tetracycline (4), ampicillin (2), and chloramphenicol (1); no strain was resistant to imipenem. Regarding virulence- and antibiotic-resistance-related genes, 19 out of 50 tested genes were present in some strains; there was a variable association of expression. Based on the obtained data, the isolates presented relatively safe characteristics and behavior, findings that should lead to further studies to assess their potential usage as beneficial cultures in the food industry.202031970700
271930.9998Antimicrobial resistance and virulence signatures of Listeria and Aeromonas species recovered from treated wastewater effluent and receiving surface water in Durban, South Africa. BACKGROUND: Treated wastewater effluent has been found to contain high levels of contaminants, including disease-causing bacteria such as Listeria and Aeromonas species. The aim of this study was to evaluate the antimicrobial resistance and virulence signatures of Listeria and Aeromonas spp. recovered from treated effluents of two wastewater treatment plants and receiving rivers in Durban, South Africa. METHODS: A total of 100 Aeromonas spp. and 78 Listeria spp. were positively identified based on biochemical tests and PCR detection of DNA region conserved in these genera. The antimicrobial resistance profiles of the isolates were determined using Kirby Bauer disc diffusion assay. The presence of important virulence genes were detected via PCR, while other virulence determinants; protease, gelatinase and haemolysin were detected using standard assays. RESULTS: Highest resistance was observed against penicillin, erythromycin and nalidixic acid, with all 78 (100%) tested Listeria spp displaying resistance, followed by ampicillin (83.33%), trimethoprim (67.95%), nitrofurantoin (64.10%) and cephalosporin (60.26%). Among Aeromonas spp., the highest resistance (100%) was observed against ampicillin, penicillin, vancomycin, clindamycin and fusidic acid, followed by cephalosporin (82%), and erythromycin (58%), with 56% of the isolates found to be resistant to naladixic acid and trimethoprim. Among Listeria spp., 26.92% were found to contain virulence genes, with 14.10, 5.12 and 21% harbouring the actA, plcA and iap genes, respectively. Of the 100 tested Aeromonas spp., 52% harboured the aerolysin (aer) virulence associated gene, while lipase (lip) virulence associated gene was also detected in 68% of the tested Aeromonas spp. CONCLUSIONS: The presence of these organisms in effluents samples following conventional wastewater treatment is worrisome as this could lead to major environmental and human health problems. This emphasizes the need for constant evaluation of the wastewater treatment effluents to ensure compliance to set guidelines.201526498595
136240.9998Distribution of phenotypic and genotypic antimicrobial resistance and virulence genes in Vibrio parahaemolyticus isolated from cultivated oysters and estuarine water. A total of 594 Vibrio parahaemolyticus isolates from cultivated oysters (n = 361) and estuarine water (n = 233) were examined for antimicrobial resistance (AMR) phenotype and genotype and virulence genes. Four hundred forty isolates (74.1%) exhibited resistance to at least one antimicrobial agent and 13.5% of the isolates were multidrug-resistant strains. Most of the V. parahaemolyticus isolates were resistant to erythromycin (54.2%), followed by sulfamethoxazole (34.7%) and trimethoprim (27.9%). The most common resistance genes were qnr (77.8%), strB (27.4%) and tet(A) (22.1%), whereas blaTEM (0.8%) was rarely found. Four isolates (0.7%) from oysters (n = 2) and estuarine water (n = 2) were positive to tdh, whereas no trh-positive isolates were observed. Significantly positive associations among AMR genes were observed. The SXT elements and class 1, 2 and 3 integrons were absent in all isolates. The results indicated that V. parahaemolyticus isolates from oysters and estuarine water were potential reservoirs of resistance determinants in the environment. This increasing threat of resistant bacteria in the environment potentially affects human health. A 'One Health' approach involved in multidisciplinary collaborations must be implemented to effectively manage antimicrobial resistance.202032358958
268650.9998Antimicrobial resistance in Escherichia coli and Salmonella spp. isolates from fresh produce and the impact to food safety. Foodborne diseases associated with fresh produce consumption have escalated worldwide, causing microbial safety of produce of critical importance. Bacteria that have increasingly been detected in fresh produce are Escherichia coli and Salmonella spp., both of which have been shown to progressively display antimicrobial resistance. The study focused on the assessment of antimicrobial resistance of these enteric bacteria from different kinds of fresh produce from various open air markets and supermarkets in the Philippines. Using the disk diffusion assay on a total of 50 bacterial isolates obtained from 410 fresh produce surveyed, monoresistance to tetracycline was observed to be the most prevalent (38%), followed by multidrug resistance to tetracycline, chloramphenicol, ciprofloxacin, and nalidixic acid (4%), and lastly by dual resistance to tetracycline and chloramphenicol (2%). Using multiplex and simplex polymerase chain reaction (PCR) assays, tetA (75%) and tetB (9%) were found in tetracycline resistant isolates, whereas catI (67%) and catIII (33%) were detected in chloramphenicol resistant isolates. Sequence analysis of gyr and par genes from the ciprofloxacin and nalidixic acid resistant isolates revealed different mutations. Based on the results, fresh produce act as a reservoir of these antibiotic resistant bacteria which may pose health threat to consumers.201728679083
269360.9998Prevalence, Antimicrobial Resistance and Toxin-Encoding Genes of Clostridioides difficile from Environmental Sources Contaminated by Feces. Clostridioides difficile (C. difficile) is the most common pathogen causing antibiotic-associated intestinal diseases in humans and some animal species, but it can also be present in various environments outside hospitals. Thus, the objective of this study was to investigate the presence and the characteristics of toxin-encoding genes and antimicrobial resistance of C. difficile isolates from different environmental sources. C. difficile was found in 32 out of 81 samples (39.50%) after selective enrichment of spore-forming bacteria and in 45 samples (55.56%) using a TaqMan-based qPCR assay. A total of 169 C. difficile isolates were recovered from those 32 C. difficile-positive environmental samples. The majority of environmental C. difficile isolates were toxigenic, with many (88.75%) positive for tcdA and tcdB. Seventy-four isolates (43.78%) were positive for binary toxins, cdtA and cdtB, and 19 isolates were non-toxigenic. All the environmental C. difficile isolates were susceptible to vancomycin and metronidazole, and most isolates were resistant to ciprofloxacin (66.86%) and clindamycin (46.15%), followed by moxifloxacin (13.02%) and tetracycline (4.73%). Seventy-five isolates (44.38%) showed resistance to at least two of the tested antimicrobials. C. difficile strains are commonly present in various environmental sources contaminated by feces and could be a potential source of community-associated C. difficile infections.202336671363
288370.9998In vitro susceptibility of Vibrio spp. isolated from the environment. Bacteria of the genus Vibrio include harmless aquatic strains as well as strains capable of causing epidemics of cholera and human intestinal diseases. Some of these species may show resistance to different antibiotics including cefotaxime, tetracycline and chloramphenicol. The susceptibility to different antibiotics was tested using 40 Vibrio alginolyticus, eight V. parahaemolyticus and six V. vulnificus strains isolated in the coastal waters of Northern Sardinia (Italy). The frequency of resistance to beta-lactams was unexpectedly high. More than 80% of Vibrio isolates were resistant to ampicillin and 2.5% of V. alginolyticus were resistant to ceftazidime and cefotetan. Forty percent of V. alginolyticus and three V. vulnificus isolates gave a positive nitrocefin test. PCR was also performed using selected primers chosen for having common sequences of bla(TEM) and bla(SHV) genes.200111337229
286180.9998Antibiotic Resistance Profiles and Genomic Analysis of Endophytic Bacteria Isolates from Wild Edible Fungi in Yunnan. The use of antibiotics has led to the emergence of antibiotic resistance, posing significant challenges in the prevention, control, and treatment of microbial diseases, while threatening public health, the environment, and food safety. In this study, the antibiotic resistance phenotypes and genotypes of 56 endophytic bacteria isolates from three species of wild edible fungi in Yunnan were analyzed using the Kirby-Bauer disk diffusion method and PCR amplification. The results revealed that all isolates were sensitive to ofloxacin, but resistance was observed against 17 other antibiotics. Specifically, 55, 53, and 51 isolates exhibited resistance to amoxicillin, penicillin, and vancomycin, respectively. Antibiotic resistance gene (ARG) detection indicated that the sulfonamide sul1 gene had the highest detection rate (53.57%). Excluding the ARG that was not detected, the lowest detection rates were the sulfonamide sul2 and sul3 genes, both at 1.79%. Among six tetracycline resistance genes, only tetK and tetM were detected. For β-lactam antibiotics, blaTEM, blaVIM, and blaSHV genes were present, while blaOXA was absent. In aminoglycoside resistance genes, aadB was not detected, while detection rates for aac(3')-IIa, acrB, and aadA1 were 3.57%, 1.79%, and 37.5%, respectively. The chloramphenicol Cat gene was detected at a rate of 14.29%, whereas floR was absent. For polypeptide resistance, VanC was detected at 3.57%, with EmgrB not detected. All three quinolone genes were detected, with detection rates of 8.92% for GyrA, 39.29% for GyrB, and 37.5% for ParC. Through phylogenetic analysis, 12 isolates that are closely related to ten common foodborne pathogenic bacteria were further selected for whole-genome sequencing and assembly. Gene annotations revealed that each isolate contained more than 15 ARGs and over 30 virulence factors. Notably, the detection rate of antibiotic resistance phenotypes was higher than that of genotypes, highlighting the importance of studying phenotypic antibiotic resistance that lacks identifiable ARGs. This study enriches the research on endophytes in wild edible fungi and provides new data for microbial ecology and antibiotic resistance research. It also offers critical insights for monitoring microbial antibiotic resistance in wild edible fungi and potentially other food sources, contributing to more effective strategies for ecological protection, sustainable agricultural development, and public health security.202540005728
554790.9998Antibiotic Resistant Bacterial Isolates from Captive Green Turtles and In Vitro Sensitivity to Bacteriophages. This study aimed to test multidrug resistant isolates from hospitalised green turtles (Chelonia mydas) and their environment in North Queensland, Australia, for in vitro susceptibility to bacteriophages. Seventy-one Gram-negative bacteria were isolated from green turtle eye swabs and water samples. Broth microdilution tests were used to determine antibiotic susceptibility. All isolates were resistant to at least two antibiotics, with 24% being resistant to seven of the eight antibiotics. Highest resistance rates were detected to enrofloxacin (77%) and ampicillin (69.2%). More than 50% resistance was also found to amoxicillin/clavulanic acid (62.5%), ceftiofur (53.8%), and erythromycin (53.3%). All the enriched phage filtrate mixtures resulted in the lysis of one or more of the multidrug resistant bacteria, including Vibrio harveyi and V. parahaemolyticus. These results indicate that antibiotic resistance is common in Gram-negative bacteria isolated from hospitalised sea turtles and their marine environment in North Queensland, supporting global concern over the rapid evolution of multidrug resistant genes in the environment. Using virulent bacteriophages as antibiotic alternatives would not only be beneficial to turtle health but also prevent further addition of multidrug resistant genes to coastal waters.201729147114
2688100.9998Intestinal and Extraintestinal Pathotypes of Escherichia coli Are Prevalent in Food Prepared and Marketed on the Streets from the Central Zone of Mexico and Exhibit a Differential Phenotype of Resistance Against Antibiotics. Background/Objectives: Antibiotic resistance is a serious public health problem threatening the treatment of infectious diseases caused by Escherichia coli, the main source of food contamination and responsible for many infectious diseases with high indices of AR profiles. Our objective was to study the presence of Escherichia coli in foods that are distributed and prepared on the street, characterizing its sensitivity profile and resistance to antibiotic drugs commonly prescribed in this geographical area. Methods: Standard procedures were performed to identify and isolate E. coli colonies from food samples collected during a three-year study. Susceptibility assays were conducted to determine the antibiotic resistance profile, and Colony PCR assays were performed to determine the pathogenic and antibiotic resistance genes. Results: A total of 189 food samples were collected, and 100% of the samples were positive for E. coli, with higher percentages of contamination for vegetables and fruits. ETEC (lt) and UPEC (vat, cnf1, hylA) genes were identified in 100% of the samples and DAEC (afa) in 27%. E. coli exhibited high percentages of resistance against ampicillin and amoxicillin/clavulanic acid (100%) and cephalexin (45%). The most effective antibiotics were tetracycline, TMP-SMX, polymyxin, and quinolones. The AR genes tetA, sul1, catA1, strA, qnrS, and floR were identified among the samples. Conclusions: Food prepared and marketed on the streets seriously threatens human health. Ampicillin and amoxicillin/clavulanic acid should not be used to treat infections caused by the multidrug-resistant ETEC and UPEC identified in this area. To our knowledge, this is the first study that explores the status of AR in this geographical area.202540298585
5558110.9998Respiratory microbiota of healthy broilers can act as reservoirs for multidrug-resistant Escherichia coli. This study aimed at evaluate the presence and to study characteristics of Escherichia coli in the respiratory system microbiota of healthy broilers. Trachea, air sacs, and lungs of 20 broilers were analyzed at 21 days of age, reared in experimental conditions, without receiving antimicrobials. E. coli strains were isolated and identified using conventional bacteriology through morphological and biochemical characterization. The production of bacteriocin-like substances, the presence of virulence-associated genes (VAGs) of APEC (Avian Pathogenic Escherichia coli) predictors, and the antimicrobial susceptibility were evaluated. E. coli was found in 85 % of the animals (17/20), in the trachea, air sacs or lungs; and it was not found in 15 % of the animals (3/20). A total of 34 isolates were recovered, 13 from the air sacs, 13 from the lungs, and 8 from the trachea, which showed no production of bacteriocin-like substances nor virulence genes associated with APEC. Most isolates, 59 % (20/34), showed resistance to at least one of the tested antimicrobials, and six multiresistant strains were identified. The results demonstrated that strains of E. coli were commensal of the respiratory microbiota, and that they did not present pathogenicity to the host, since there were no clinical signs of disease, macroscopic lesions in the organs of the evaluated broilers, production of bacteriocin-like substances, nor virulence-associated genes considered as predictors of APEC in bacteria. These strains of E. coli were mostly susceptible to antimicrobials. However, the occurrence of multidrug-resistant strains suggests that these animals can act as reservoirs of resistant to antimicrobials E. coli.202134507109
2788120.9998Investigating the virulence genes and antibiotic susceptibility patterns of Vibrio cholerae O1 in environmental and clinical isolates in Accra, Ghana. BACKGROUND: Cholera has been endemic in Ghana since its detection in 1970. It has been shown that long-term survival of the bacteria may be attained in aquatic environments. Consequently, cholera outbreaks may be triggered predominantly in densely populated urban areas. We investigated clinical and environmental isolates of Vibrio cholerae O1 in Accra to determine their virulence genes, antibiotic susceptibility patterns and environmental factors maintaining their persistence in the environment. METHODS: Water samples from various sources were analyzed for the presence of V. cholerae O1 using culture methods. Forty clinical isolates from a previous cholera outbreak were included in the study for comparison. Antibiotic susceptibility patterns of the bacteria were determined by disc diffusion. Virulence genes were identified by analyzing genes for ctx, tcpA (tcpA(El Tor) tcpA(Cl)), zot, ompW, rbfO1 and attRS using PCR. Physicochemical characteristics of water were investigated using standard methods. One-way ANOVA and student t - test were employed to analyze the relationship between physicochemical factors and the occurrence of V. cholerae O1. RESULTS: Eleven V. cholerae O1 strains were successfully isolated from streams, storage tanks and wells during the study period. All isolates were resistant to one or more of the eight antibiotics used. Multidrug resistance was observed in over 97% of the isolates. All isolates had genes for at least one virulence factor. Vibrio cholerae toxin gene was detected in 82.4% of the isolates. Approximately 81.8% of the isolates were positive for tcpA(El Tor) gene, but also harbored the tcpA(cl) gene. Isolates were grouped into thirteen genotypes based on the genes analyzed. High temperature, salinity, total dissolved solids and conductivity was found to significantly correlate positively with isolation of V. cholerae O1. V. cholerae serotype Ogawa biotype El tor is the main biotype circulating in Ghana with the emergence of a hybrid strain. CONCLUSIONS: Multidrug resistant V. cholerae O1 with different genotypes and pathogenicity are present in water sources and co-exist with non O1/O139 in the study area.201930665342
2700130.9998Prevalence of Salmonella Typhimurium and Salmonella Enteritidis isolated from poultry meat: virulence and antimicrobial-resistant genes. Salmonellosis, a zoonotic disease, is one of the leading causes of foodborne illness worldwide. It is responsible for most infections caused by consumption of contaminated food. In recent years, a significant increase in the resistance of these bacteria to common antibiotics has been observed, posing a serious threat to global public health. The aim of this study was to investigate the prevalence of virulent antibiotic-resistant Salmonella spp. strains in Iranian poultry markets. A total of 440 chicken meat samples were randomly selected from meat supply and distribution facilities in Shahrekord and tested for bacteriological contamination. After culturing and isolating the strains, identification was performed using the classical bacteriological method and PCR. To determine antibiotic resistance, a disc diffusion test was performed according to the recommendations of the French Society of Microbiology. PCR was used to detect resistance and virulence genes. Only 9% of the samples were positive for Salmonella. These were Salmonella typhimurium isolates. All Salmonella typhimurium serotypes tested positive for the rfbJ, fljB, invA and fliC genes. Resistance to TET, cotrimoxazole, NA, NIT, piperacillin/tazobactam and other antibiotics was found in 26 (72.2%), 24 (66.7%), 22 (61.1%) and 21 (58.3%) isolates, respectively. The sul1, sul2 and sul3 genes were present in 20, 12 and 4 of 24 cotrimoxazole-resistant bacteria, respectively. Chloramphenicol resistance was found in six isolates, but more isolates tested positive for the floR and cat two genes. In contrast, 2 (33%) of the cat three genes, 3 (50%) of the cmlA genes and 2 (34%) of the cmlB genes were all positive. The results of this investigation showed that Salmonella typhimurium is the most common serotype of the bacterium. This means that most of the antibiotics commonly used in the livestock and poultry industries are ineffective against most Salmonella isolates, which is important for public health.202337322421
2706140.9998Prevalence 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
2677150.9997Detection of Staphylococcus Isolates and Their Antimicrobial Resistance Profiles and Virulence Genes from Subclinical Mastitis Cattle Milk Using MALDI-TOF MS, PCR and Sequencing in Free State Province, South Africa. Staphylococcus species are amongst the bacteria that cause bovine mastitis worldwide, whereby they produce a wide range of protein toxins, virulence factors, and antimicrobial-resistant properties which are enhancing the pathogenicity of these organisms. This study aimed to detect Staphylococcus spp. from the milk of cattle with subclinical mastitis using MALDI-TOF MS and 16S rRNA PCR as well as screening for antimicrobial resistance (AMR) and virulence genes. Our results uncovered that from 166 sampled cows, only 33.13% had subclinical mastitis after initial screening, while the quarter-level prevalence was 54%. Of the 50 cultured bacterial isolates, MALDI-TOF MS and 16S rRNA PCR assay and sequencing identified S. aureus as the dominant bacteria by 76%. Furthermore, an AMR susceptibility test showed that 86% of the isolates were resistant to penicillin, followed by ciprofloxacin (80%) and cefoxitin (52%). Antimicrobial resistance and virulence genes showed that 16% of the isolates carried the mecA gene, while 52% of the isolates carried the Lg G-binding region gene, followed by coa (42%), spa (40%), hla (38%), and hlb (38%), whereas sea and bap genes were detected in 10% and 2% of the isolates, respectively. The occurrence of virulence factors and antimicrobial resistance profiles highlights the need for appropriate strategies to control the spread of these pathogens.202438200885
2909160.9997Determination of the prevalence of antimicrobial resistance genes in canine Clostridium perfringens isolates. Clostridium perfringens is a well documented cause of a mild self-limiting diarrhea and a potentially fatal acute hemorrhagic diarrheal syndrome in the dog. A recent study documented that 21% of canine C. perfringens isolates had MIC's indicative of resistance to tetracycline, an antimicrobial commonly recommended for treatment of C. perfringens-associated diarrhea. The objective of the present study was to further evaluate the antimicrobial susceptibility profiles of these isolates by determining the prevalence of specific resistance genes, their expression, and ability for transference between bacteria. One hundred and twenty-four canine C. perfringens isolates from 124 dogs were evaluated. Minimum inhibitory concentrations of tetracycline, erythromycin, tylosin, and metronidazole were determined using the CLSI Reference Agar Dilution Method. All isolates were screened for three tetracycline resistance genes: tetA(P), tetB(P) and tetM, and two macrolide resistance genes: ermB and ermQ, via PCR using primer sequences previously described. Ninety-six percent (119/124) of the isolates were positive for the tetA(P) gene, and 41% (51/124) were positive for both the tetA(P) and tetB(P) genes. No isolates were positive for the tetB(P) gene alone. Highly susceptible isolates (MIC< or = 4 microg/ml) were significantly more likely to lack the tetB(P) gene. One isolate (0.8%) was positive for the ermB gene, and one isolate was positive for the ermQ gene. The tetM gene was not found in any of the isolates tested. Two out of 15 tested isolates (13%) demonstrated transfer of tetracycline resistance via bacterial conjugation. Tetracycline should be avoided for the treatment of C. perfringens-associated diarrhea in dogs because of the relatively high prevalence of in vitro resistance, and the potential for conjugative transfer of antimicrobial resistance.200616330169
2927170.9997Aeromonas hydrophila from marketed mullet (Mugil cephalus) in Egypt: PCR characterization of β-lactam resistance and virulence genes. AIMS: Aeromonas hydrophila has been isolated from various fish species in Egypt and is known to carry virulence and antimicrobial resistance genes, which pose a risk for public health. The aim of the present study is to report, for the first time, the infection of mullet (Mugil cephalus) with A. hydrophila and to clarify the potential association between antimicrobial resistance and virulence traits encoded in A. hydrophila. METHODS AND RESULTS: In this study, the occurrence of A. hydrophila in marketed mullet and the antimicrobial resistance phenotypes of these isolates were determined. Aeromonas hydrophila isolates were screened for the presence of virulence and β-lactam resistance genes; the correlation between both gene groups was also investigated. The infection rate of examined mullet with A. hydrophila was 37% (50/135). The highest antimicrobial resistance was detected to cefoxitin (100%), followed by ampicillin (84%), ceftazidime (56%) and cefotaxime (40%). Only 4% of the isolates were resistant to erythromycin; 6% were resistant to both gentamicin and kanamycin with no resistance to ciprofloxacin. Variable frequencies of virulence and β-lactam resistance genes were evident from PCR, where aerA and bla(TEM) predominated. The study also indicated a general weak positive correlation (R = 0·3) between both virulence and β-lactam resistance genes. Some of the studied virulence genes (e.g. aerA:hlyA and hlyA:ast) were found to correlate positively. CONCLUSIONS: The presence of virulence and resistance genes in A. hydrophila from food sources poses a serious threat to public health. To our knowledge, this is the first report describing the occurrence of A. hydrophila in mullet and highlighting the coexistence of virulence and β-lactam resistance genes encoded by these bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: These data provide insights into the potential association of antimicrobial resistance and virulence genes in A. hydrophila from marketed mullet in Egypt, which could pose threats to humans even if a weak positive correlation exists between both genes.201829453863
2903180.9997Soil Bacteria in Urban Community Gardens Have the Potential to Disseminate Antimicrobial Resistance Through Horizontal Gene Transfer. Fifteen soil and 45 vegetable samples from Detroit community gardens were analyzed for potential antimicrobial resistance contamination. Soil bacteria were isolated and tested by antimicrobial susceptibility profiling, horizontal gene transfer, and whole-genome sequencing. High-throughput 16S rRNA sequencing analysis was conducted on collected soil samples to determine the total bacterial composition. Of 226 bacterial isolates recovered, 54 were from soil and 172 from vegetables. A high minimal inhibitory concentration (MIC) was defined as the MIC greater than or equal to the resistance breakpoint of Escherichia coli for Gram-negative bacteria or Staphylococcus aureus for Gram-positive bacteria. The high MIC was observed in 63.4 and 69.8% of Gram-negative isolates from soil and vegetables, respectively, against amoxicillin/clavulanic acid, as well as 97.5 and 82.7% against ampicillin, 97.6 and 90.7% against ceftriaxone, 85.4 and 81.3% against cefoxitin, 65.8 and 70.5% against chloramphenicol, and 80.5 and 59.7% against ciprofloxacin. All Gram-positive bacteria showed a high MIC to gentamicin, kanamycin, and penicillin. Forty of 57 isolates carrying tetM (70.2%) successfully transferred tetracycline resistance to a susceptible recipient via conjugation. Whole-genome sequencing analysis identified a wide array of antimicrobial resistance genes (ARGs), including those encoding AdeIJK, Mex, and SmeDEF efflux pumps, suggesting a high potential of the isolates to become antimicrobial resistant, despite some inconsistency between the gene profile and the resistance phenotype. In conclusion, soil bacteria in urban community gardens can serve as a reservoir of antimicrobial resistance with the potential to transfer to clinically important pathogens, resulting in food safety and public health concerns.202134887843
5579190.9997Survey of Shiga toxigenic Escherichia coli O157 and drug-resistant coliform bacteria from in-line milk filters on dairy farms in the Czech Republic. AIMS: To determine the occurrence of Shiga toxin-producing Escherichia coli (STEC) O157 and coliform bacteria isolates resistant to antimicrobial agents in dairy herds by examining milk filters and to analyse the influence of management factors and antibiotic use on antimicrobial resistance. METHODS AND RESULTS: A total of 192 in-line milk filters were sampled on 192 dairy farms in the Czech Republic. Information on feeding, husbandry, production, and antibiotic therapy were obtained by questionnaire. The milk filters were cultured for STEC O157 and coliform bacteria. All recovered isolates were examined for antimicrobial susceptibility and presence of antimicrobial-resistance genes. STEC O157 was detected in four (2%) of the filters. Resistant nonpathogenic E. coli and coliform bacteria isolates with specific genes were detected in 44 (23%) of the filters. CONCLUSIONS: The study demonstrated a high prevalence of resistant coliform bacteria in milk filters obtained on Czech dairy farms. SIGNIFICANCE AND IMPACT OF THE STUDY: The occurrence of resistant coliform bacteria in milk filters was significantly higher among isolates from farms where antibiotic therapy against mastitis was employed during the dry period (P < 0.05).200817953684