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131300.9996Molecular epidemiology of ceftiofur-resistant Escherichia coli isolates from dairy calves. Healthy calves (n = 96, 1 to 9 weeks old) from a dairy herd in central Pennsylvania were examined each month over a five-month period for fecal shedding of ceftiofur-resistant gram-negative bacteria. Ceftiofur-resistant Escherichia coli isolates (n = 122) were characterized by antimicrobial resistance (disk diffusion and MIC), serotype, pulsed-field gel electrophoresis subtypes, beta-lactamase genes, and virulence genes. Antibiotic disk diffusion assays showed that the isolates were resistant to ampicillin (100%), ceftiofur (100%), chloramphenicol (94%), florfenicol (93%), gentamicin (89%), spectinomycin (72%), tetracycline (98%), ticarcillin (99%), and ticarcillin-clavulanic acid (99%). All isolates were multidrug resistant and displayed elevated MICs. The E. coli isolates belonged to 42 serotypes, of which O8:H25 was the predominant serotype (49.2%). Pulsed-field gel electrophoresis classified the E. coli isolates into 27 profiles. Cluster analysis showed that 77 isolates (63.1%) belonged to one unique group. The prevalence of pathogenic E. coli was low (8%). A total of 117 ceftiofur-resistant E. coli isolates (96%) possessed the bla(CMY2) gene. Based on phenotypic and genotypic characterization, the ceftiofur-resistant E. coli isolates belonged to 59 clonal types. There was no significant relationship between calf age and clonal type. The findings of this study revealed that healthy dairy calves were rapidly colonized by antibiotic-resistant strains of E. coli shortly after birth. The high prevalence of multidrug-resistant nonpathogenic E. coli in calves could be a significant source of resistance genes to other bacteria that share the same environment.200616751500
104410.9996Molecular Characterization of Antimicrobial Resistance and Virulence Genes of Bacterial Pathogens from Bovine and Caprine Mastitis in Northern Lebanon. Mastitis is an infectious disease encountered in dairy animals worldwide that is currently a growing concern in Lebanon. This study aimed at investigating the etiology of the main mastitis-causing pathogens in Northern Lebanon, determining their antimicrobial susceptibility profiles, and identifying their antimicrobial resistance (AMR) genes. A total of 101 quarter milk samples were collected from 77 cows and 11 goats presenting symptoms of mastitis on 45 dairy farms. Bacterial identification was carried out through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibility was tested by disc diffusion and broth microdilution methods. Molecular characterization included polymerase chain reaction (PCR) screening for genes encoding extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC among Enterobacterales isolates, and virulence factors among Staphylococcus isolates. Escherichia coli isolates were subjected to phylogenetic typing by a quadruplex PCR method. The most frequently identified species were Streptococcus uberis (19.2%), Streptococcus agalactiae (15.1%), E. coli (12.3%), and Staphylococcus aureus (10.96%). Gram-positive bacteria were resistant to macrolides and tetracycline, whereas gram-negative bacteria displayed resistance to ampicillin and tetracycline. Two ESBL genes, bla(TEM) (83.3%) and bla(OXA) (16.7%), and one AmpC beta-lactamase gene, bla(CMY-II) (16.7%), were detected among six E. coli isolates, which mainly belonged to phylogenetic group B1. Among Staphylococcus spp., the mecA gene was present in three isolates. Furthermore, four isolates contained at least one toxin gene, and all S. aureus isolates carried the ica operon. These findings revealed the alarming risk of AMR in the Lebanese dairy chain and the importance of monitoring antimicrobial usage.202134071800
112520.9996Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal. AIM: The aim of this work was to detect antibiotic resistance in Gram-negative bacteria isolated from subclinical mastitis in cattle in West Bengal. MATERIALS AND METHODS: The milk samples were collected from the cattle suffering with subclinical mastitis in West Bengal. The milk samples were inoculated into the nutrient broth and incubated at 37°C. On the next day, the growth was transferred into nutrient agar and MacConkey agar. All the pure cultures obtained from nutrient agar slant were subjected to Gram-staining and standard biochemical tests. All the bacterial isolates were tested in vitro for their sensitivity to different antibiotics commonly used in veterinary practices. All Gram-negative isolates including positive control were subjected to polymerase chain reaction (PCR) for detection of bla(CTX-M), bla(TEM), bla(SHV), bla(VIM), tetA, tetB, tetC, and tetM genes considered for extended-spectrum β-lactamase (ESBL), metallo-β-lactamase, and tetracycline resistance. RESULTS: In total, 50 Gram-negative organisms (Escherichia coli, Proteus, Pseudomonas, Klebsiella, and Enterobacter) were isolated from milk samples of subclinical mastitis infected cattle. Among these Gram-negative isolates, 48% (24/50) were found either ESBL producing or tetracycline resistant. Out of total 50 Gram-negative isolates, bla(CTX-M) was detected in 18 (36%) isolates, and 6 (12%) harbored bla(TEM) genes in PCR. None of the isolates carried bla(SHV) genes. Further, in this study, 5 (10%) isolates harbored tet(A) gene, and 8 (16%) isolates carried tet(B) gene. No tet(C) gene was detected from the isolates. CONCLUSION: This study showed emerging trend of antibiotic-resistant Gram-negative bacteria associated with subclinical mastitis in cattle in West Bengal, India.201728620255
214230.9996Resistance to β-lactams and distribution of β-lactam resistance genes in subgingival microbiota from Spanish patients with periodontitis. OBJECTIVES: The aim of this study was to analyze the distribution of β-lactamase genes and the multidrug resistance profiles in β-lactam-resistant subgingival bacteria from patients with periodontitis. MATERIALS AND METHODS: Subgingival samples were obtained from 130 Spanish patients with generalized periodontitis stage III or IV. Samples were grown on agar plates with amoxicillin or cefotaxime and incubated in anaerobic and microaerophilic conditions. Isolates were identified to the species level by the sequencing of their 16S rRNA gene. A screening for the following β-lactamase genes was performed by the polymerase chain reaction (PCR) technique: bla(TEM), bla(SHV), bla(CTX-M), bla(CfxA), bla(CepA), bla(CblA), and bla(ampC). Additionally, multidrug resistance to tetracycline, chloramphenicol, streptomycin, erythromycin, and kanamycin was assessed, growing the isolates on agar plates with breakpoint concentrations of each antimicrobial. RESULTS: β-lactam-resistant isolates were found in 83% of the patients. Seven hundred and thirty-seven isolates from 35 different genera were obtained, with Prevotella and Streptococcus being the most identified genera. bla(CfxA) was the gene most detected, being observed in 24.8% of the isolates, followed by bla(TEM) (12.9%). Most of the isolates (81.3%) were multidrug-resistant. CONCLUSIONS: This study shows that β-lactam resistance is widespread among Spanish patients with periodontitis. Furthermore, it suggests that the subgingival commensal microbiota might be a reservoir of multidrug resistance and β-lactamase genes. CLINICAL RELEVANCE: Most of the samples yielded β-lactam-resistant isolates, and 4 different groups of bla genes were detected among the isolates. Most of the isolates were also multidrug-resistant. The results show that, although β-lactams may still be effective, their future might be hindered by the presence of β-lactam-resistant bacteria and the presence of transferable bla genes.202032495224
266740.9996Prevalence, virulence and antimicrobial resistance patterns of Aeromonas spp. isolated from children with diarrhea. BACKGROUND: Aeromonas spp. cause various intestinal and extraintestinal diseases. These bacteria are usually isolated from fecal samples, especially in children under five years old. The aim of this study was to assess the prevalence of Aeromonas spp. and their antimicrobial resistance profile in children with diarrhea referred to the Children Medical Center in Tehran, between 2013 and 2014. METHODS: A total number of 391 stool samples were collected from children with ages between 1 day and 14 years old, with diarrhea (acute or chronic), referred to the Children Hospital, Tehran, Iran, between 2013 and 2014. Samples were enriched in alkaline peptone water broth for 24 hours at 37 °C and then cultured. Suspicious colonies were analyzed through biochemical tests. Furthermore, antimicrobial susceptibility tests were carried out for the isolates. Isolates were further studied for act, ast, alt, aerA and hlyA virulence genes using polymerase chain reaction. RESULTS: In total, 12 isolates (3.1%) were identified as Aeromonas spp.; all were confirmed using the API-20E test. Of these isolates, five A. caviae (42%), four A. veronii (33%) and three A. hydrophila (25%) were identified in cases with gastroenteritis. Second to ampicillin (which was included in the growth medium used), the highest rate of antimicrobial resistance was seen against nalidixic acid and trimethoprim-sulfamethoxazole (5 isolates each, 41.6%) and the lowest rate of antimicrobial resistance was seen against gentamicin, amikacin and cefepime (none of the isolates). Results included 76.4% act, 64.7% ast, 71.5% alt, 83.3% aerA and 11.7% hlyA genes. CONCLUSION: Aeromonas spp. are important due to their role in diarrhea in children; therefore, isolation and identification of these fecal pathogens should seriously be considered in medical laboratories. Since virulence genes play a significant role in gastroenteritis symptoms caused by these bacteria, Aeromonas species that include virulence genes are potentially suspected to cause severe infections. Moreover, bacterial antimicrobial resistance is increasing, especially against trimethoprim-sulfamethoxazole and nalidixic acid.201627622161
267950.9995Detection and Molecular Characterization of Staphylococci from Eggs of Household Chickens. Eggs are a healthy and nutritious food source, but may be contaminated by bacteria. Previous studies have reported the presence of staphylococci in eggs of farmed chickens, but no study has evaluated the staphylococcal population of eggs from household chickens. In this study, staphylococci from eggs (n = 275) of household chickens collected from November 2016 to March 2017 from different villages of Khyber Pakhtunkhwa province, Pakistan, were characterized. Seven species of staphylococci were identified from 65 eggs, including the predominant species, Staphylococcus xylosus (49/275; 17.8%). S. xylosus isolates (n = 73) were tested for antimicrobial susceptibility, presence of resistance genes, genetic relatedness, and inhibitory activity against other bacteria. The majority of isolates were resistant to oxacillin (83.6%) and tetracycline (24.7%), but also exhibited resistance to daptomycin and linezolid (5.5% each). Of the 10 resistance genes tested, isolates were only positive for mecA (35.6%; 26/73), mecC/C1 (2.7%; 2/73), and tet(K) (14/73; 19%). Using pulsed-field gel electrophoresis (PFGE), nine clusters had identical PFGE patterns. Isolates produced inhibitory activity against a broad spectrum of bacteria; 20.5%, 19.2%, 17.8%, and 16.4% of S. xylosus were able to inhibit growth of Salmonella enterica serotype Typhi, methicillin-susceptible Staphylococcus aureus, Escherichia coli, and methicillin-resistant Staphylococcus aureus, respectively. This study demonstrated the presence of genetically related antimicrobial-resistant S. xylosus from eggs from household chickens. Like table eggs, eggs of household chickens also contain staphylococci that may be resistant to antimicrobials used to treat human infections. These data will allow comparison between staphylococci from eggs from different sources and may indicate the relative safety of eggs from household chickens. Further study of these egg types and their microbial composition is warranted.201931009262
116460.9995The distribution of beta lactamase genes in Escherichia coli phylotypes isolated from diarrhea and UTI cases in northwest Iran. BACKGROUND: Pathogenic Escherichia coli strains are a common cause of intestinal and extra-intestinal infections, especially in developing countries. Extended spectrum beta-lactamases (ESBLS), a heterogeneous group of plasmid-encoded beta-lactamases, are common throughout the world. OBJECTIVES: The aim of the present study was to determine the phenotypic and genotypic characteristics of ESBLS produced by E. coli isolates taken from patients with diarrhea and urinary tract infections (UTI) in northwest Iran. MATERIAL AND METHODS: A total of 132 E. coli isolates (92 isolates from UTI and 40 isolates from diarrheic cases) were recovered and confirmed by biochemical tests. The isolates were examined for blaTEM and blaSHV genes and phylogenetic background by two multiplex PCR assays. The isolates were tested for antibiotic susceptibility against nine antibiotic agents by the disk diffusion method. RESULTS: The phylogenetic analysis showed that the UTI isolates mostly fell into phylo-group B2, followed by D, while the diarrheic isolates belonged to phylo-groups D and A. Out of 92 UTI isolates, 29.3% and 17.4% possessed blaTEM and blaSHV genes, respectively. Ten diarrheic isolates were positive for blaTEM, two isolates possessed the blaSHV gene, and one isolate was positive for both genes. The UTI isolates that were positive for blaTEM and blaSHV genes mostly belonged to phylo-groups D and B2, whereas the diarrhea isolates were in phylo-groups D and A. Phylogenetic group D isolates have an accumulation of ESBLS genes in the diarrheic and UTI isolates. In both the UTI and diarrhea isolates, the maximum rate of resistance was against cefazolin, and the minimum rate of resistance was against nitrofurantoin. Twenty-four antibiotic resistance patterns were observed among the isolates. The amikacin, ciprofloxacin, cefotaxime, cefuroxime, cefazolin, gentamicin, nalidixic acid and trimethoprim/sulfamethoxazole resistance pattern was the most prevalent in the isolates that belonged to phylo-group D. CONCLUSIONS: The correct choice of effective antibiotic policy is needed to limit the spread of antibiotic resistance in bacteria.201425166436
104370.9995Antibiotic Susceptibility Profiles of Bacterial Isolates Recovered from Abscesses in Cattle and Sheep at a Slaughterhouse in Algeria. Abscesses represent the most prominent emerging problem in the red meat industry, leading to great economic constraints and public health hazards. Data on etiological agents present in these purulent lesions in Algeria are very scarce. The aim of this study was to identify the bacteria responsible for these abscesses and to determine their antibiotic susceptibility profiles. A total of 123 samples of abscesses from 100 slaughtered sheep and 23 slaughtered cattle were cultured in several media. A total of 114 bacterial isolates were cultured from 103 abscesses. Bacteria were identified using MALDI-TOF, and antibiotic susceptibility was determined by the disk diffusion method on Mueller-Hinton agar. A total of 73.6% (n = 84) corresponded to Enterobacterales, of which four were multidrug-resistant (MDR). These isolates, together with Staphylococcus aureus, coagulase negative Staphylococci, and seven randomly chosen susceptible Escherichia coli isolates, were further characterized using WGS. Resistome analysis of the four MDR Enterobacterales isolates revealed the presence of OXA-48 carbapenemase in two Klebsiella pneumoniae ST985 and one E. coli ST10 isolates and a CTX-M-15 ESBL in one E. coli isolate ST1706. Two coagulase-negative Staphylococci isolates were found to carry the mecA gene. WGS showed the presence of different resistance genes and virulence genes. Our study revealed 5% of MDR Enterobacterales (including ESBLs and carbapenemases) identified from abscesses, thus urging the need for abscess monitoring in slaughterhouses.202438543576
116580.9995Isolation, Antimicrobial Susceptibility Profile and Detection of Sul1, blaTEM, and blaSHV in Amoxicillin-Clavulanate-Resistant Bacteria Isolated From Retail Sausages in Kampar, Malaysia. BACKGROUND: Due to the overuse of antibiotics in livestock as a growth-promoting agent, the emergence of multi-antibiotic resistant bacteria is becoming a concern. OBJECTIVES: In this study, we aimed to detect the presence and discover the molecular determinants of foodborne bacteria in retail sausages resistant towards the antibacterial agent amoxicillin-clavulanate. METHODS: Two grams of sausages were chopped into small pieces and transferred into sterile Luria-Bertani (LB) enrichment broths overnight before they were plated on MacConkey agar petri dishes. The bacteria isolated were then screened for amoxicillin-clavulanate resistance, and an antimicrobial susceptibility test of each isolate was performed by using the disc diffusion method. Double synergy and phenotypic tests were carried out to detect the presence of extended spectrum β-lactamase (ESBL). API 20E kit was used to identify the Enterobacteriaceae. All isolates were further examined by polymerase chain reaction (PCR) for resistant genes blaOXA-1, blaOXA-10, plasmid-mediated AmpC (blaCMY and blaDHA), and the chromosome-mediated AmpC, Sul1, blaTEM, and blaSHV genes. RESULTS: A total of 18 amoxicillin-clavulanate resistant isolates were obtained from seven different types of retail sausages. Only half of them were identified as Enterobacteriaceae, but none were ESBL-producers. All the 18 isolated strains demonstrated resistance towards amoxicillin-clavulanate, penicillin and oxacillin (100%), cefotaxime (71.4%), cefpodoxime (66.7%), and ampicillin (83.3%). blaTEM was the most frequently detected β-lactamase gene. Both plasmid- and chromosomal-bound blaTEM genes were detected in all of the isolated Enterobacteriaceae. blaSHV and Sul1 accounted for 22.2% and 11.1% of the amoxicillin-clavulanate resistant isolates, respectively, whereas blaAMPC, blaCMY, blaDHA, blaOXA-1, and blaOXA-10 were not found in any of the isolates. The only one ESBL-producing bacteria detected in this study was Chryseobacterium meningosepticum, which harbored the blaTEM gene. CONCLUSIONS: The multidrug resistant bacteria that carry antibiotic resistant genes from retail sausages may increase the risk of transmission to humans via the consumption of contaminated sausages. Stricter measures must be taken to address the use of antibiotics in animal agriculture and to consider their potential impact on human health.201627942365
214390.9995Detection of cfxA2, cfxA3, and cfxA6 genes in beta-lactamase producing oral anaerobes. Purpose The aim of this study was to identify β-lactamase-producing oral anaerobic bacteria and screen them for the presence of cfxA and BlaTEM genes that are responsible for β-lactamase production and resistance to β-lactam antibiotics. Material and Methods Periodontal pocket debris samples were collected from 48 patients with chronic periodontitis and anaerobically cultured on blood agar plates with and without β-lactam antibiotics. Presumptive β-lactamase-producing isolates were evaluated for definite β-lactamase production using the nitrocefin slide method and identified using the API Rapid 32A system. Antimicrobial susceptibility was performed using disc diffusion and microbroth dilution tests as described by CLSI Methods. Isolates were screened for the presence of the β-lactamase-TEM (BlaTEM) and β-lactamase-cfxA genes using Polymerase Chain Reaction (PCR). Amplified PCR products were sequenced and the cfxA gene was characterized using Genbank databases. Results Seventy five percent of patients carried two species of β-lactamase-producing anaerobic bacteria that comprised 9.4% of the total number of cultivable bacteria. Fifty one percent of β-lactamase-producing strains mainly Prevotella, Porphyromonas, and Bacteroides carried the cfxA gene, whereas none of them carried blaTEM. Further characterization of the cfxA gene showed that 76.7% of these strains carried the cfxA2 gene, 14% carried cfxA3, and 9.3% carried cfxA6. The cfxA6 gene was present in three Prevotella spp. and in one Porphyromonas spp. Strains containing cfxA genes (56%) were resistant to the β-lactam antibiotics. Conclusion This study indicates that there is a high prevalence of the cfxA gene in β-lactamase-producing anaerobic oral bacteria, which may lead to drug resistance and treatment failure.201627119762
1131100.9995Antimicrobial resistance genotypes and phenotypes from multidrug-resistant bacterial wound infection isolates in Cambodia. This study aimed to identify the molecular determinants responsible for antibiotic resistance among human wound isolates in Cambodia. Staphylococcus spp. (n=10) and a variety of Gram-negative isolates (n=21) were taken from a larger collection of wound isolates collected during 2011-2013 and were analysed for the presence of >230 resistance determinants using a broad-spectrum DNA microarray. These isolates were chosen to represent the species most commonly found in wound isolates referred during this time and to include some of the most resistant strains. Resistance determinants detected among the staphylococci included blaZ (90%), mecA (100%), erm(B) (70%), erm(C) (20%), tet(38) (90%), tet(K) (40%), tet(L(p)) (10%), tet(M) (20%), lnu(A)/lin(A) and lnu(B)/lin(B) (10% each), msr(A)/msr(B)/msr(SA) (10%), norA (80%) and dfrA (10%). Eleven different β-lactamase genes were detected among the Gram-negative bacteria, including genes encoding the TEM (48%), CTX-M-1 (48%), CTX-M-9 (5%), SHV (5%) and VEB (10%) families of broad-spectrum and extended-spectrum β-lactamase enzymes, as well as the carbapenemase gene bla(OXA-23). Forty additional genes were also detected in the Gram-negative isolates conferring resistance to aminoglycosides (11 genes), phenicols (5 genes), macrolides [4 genes, including mph(A)/mph(K) (10%)], lincosamides [lnu(F)/lin(F), lnu(G)/lin(G)], tetracycline (4 genes), rifampicin [arr (29%)], quaternary amines [qacEΔ1 (43%)], quinolones [qnrS (14%) and qnrB (5%)], sulfonamides [sul1 (29%), sul2 (38%) and sul3 (10%)], streptothricin (sat2) and trimethoprim (6 genes). The results obtained here provide a snapshot of the broad variety of resistance determinants currently circulating within Cambodia.201527873709
2674110.9995Phylogeny, virulence factors and antimicrobial susceptibility of Escherichia coli isolated in clinical bovine mastitis. The aim of this study was to identify specific phylogeny groups, virulence genes or antimicrobial resistance traits of Escherichia coli isolated in bovine mastitis associated to clinical signs, persistence of intramammary infection in the quarter and recovery from mastitis. A total of 154 E. coli isolates from bovine clinical mastitis, 144 from the acute stage and 10 from follow-up samples 3 weeks later, originating from 144 cows in 65 dairy herds in Southern Finland were investigated. Phylogeny groups and virulence genes of the isolates were determined using polymerase chain reaction, and antimicrobial susceptibility using the VetMIC™ microdilution method. In ten cows (11.8%), infection persisted, confirmed by re-isolation of the same pulsed-field gel electrophoresis type from the affected quarter at 3 weeks post-treatment. The majority of isolates, 119 (82.6%), belonged to phylogeny group A, which mainly consisted of commensal strains. Altogether 56 isolates (38.9%) had at least one virulence gene detected. Most common virulence genes detected were irp2, iucD, papC iss; genes svg, stx1, stx2, cnf1 and hlyA were not found. Combinations of virulence genes varied greatly. Forty (27.8%) of the 144 E. coli isolates showed resistance to at least one antimicrobial tested. None of the studied phylogeny groups, virulence factors or antimicrobial resistance traits was associated with clinical signs, persistence of intramammary infection or clinical recovery from mastitis. The results support the conclusion that mastitis-causing E. coli bacteria are typical commensals.201120729012
1266120.9995Characterization of methicillin-resistant coagulase-negative staphylococci in milk from cows with mastitis in Brazil. Staphylococci are one of the most prevalent microorganisms in bovine mastitis. Staphylococcus spp. are widespread in the environment, and can infect animals and humans as opportunistic pathogens. The objective of this study was to determine the frequency of methicillin-resistance (MR) among coagulase-negative staphylococci (CoNS) previously obtained from milk of mastitic cows in Brazil and to characterize the antimicrobial resistance phenotype/genotype and the SCCmec type of MRCoNS isolates. Identification of MRCoNS was based on both biochemical and molecular methods. Susceptibility testing for eleven antimicrobials was performed by disk-diffusion agar. Antimicrobial resistance genes and SCCmec were investigated by specific PCRs. Twenty-six MRCoNS were detected (20 % of total CoNS), obtained from 24 animals, and were identified as follows: S. epidermidis (7 isolates), S. chromogenes (7), S. warneri (6), S. hyicus (5) and S. simulans (1). All MRCoNS isolates carried mecA while the mecC gene was not detected in any CoNS. The SCCmec IVa was demonstrated in nine MRCoNS, while the remaining 17 isolates harbored non-typeable SCCmec cassettes. In addition to oxacillin and cefoxitin resistance, MRCoNS showed resistance to tetracycline (n = 7), streptomycin (n = 6), tobramycin (n = 6), and gentamicin (n = 4), and harbored the genes tet(K) (n = 7), str (n = 3), ant(4') (n = 6) and aac(6')-aph(2″) (n = 4), respectively. In addition, seven strains showed intermediate resistance to clindamycin and two to streptomycin, of which two harboured the lnu(B) and lsa(E) genes and two the aad(E) gene, respectively. One isolate presented intermediate erythromycin and clindamycin resistance and harbored an erm(C) gene with an uncommon 89-bp deletion rendering a premature stop codon. MRCoNS can be implicated in mastitis of cows and they constitute a reservoir of resistance genes that can be transferred to other pathogenic bacteria.201424817534
2972130.9995Genetic characterisation of class 1 integrons among multidrug-resistant Salmonella serotypes in broiler chicken farms. OBJECTIVES: Antimicrobial resistance in Salmonella serotypes has been reported. Integrons play an important role in the dissemination of antimicrobial resistance genes in bacteria. Scarce literature is available on the identification of integrons in Salmonella isolated from broiler chickens. In this study, antimicrobial susceptibility testing and characterisation of class 1 integrons among multidrug-resistant (MDR) Salmonella enterica serotypes in broiler chicken farms in Egypt were performed. METHODS: Antimicrobial susceptibility was determined by the disk diffusion method. PCR was performed to detect antimicrobial resistance genes and class 1 integrons in the tested Salmonella serotypes. Gene sequencing of the variable region of a class 1 integron was performed. RESULTS: Salmonella spp. were detected in 26 (13.5%) of 192 broiler samples, with Salmonella Enteritidis being the most frequently detected serotype, followed by Salmonella Kentucky and Salmonella Typhimurium and other serotypes. A very high resistance rate was observed to trimethoprim/sulfamethoxazole (100%), whilst a low resistance rate was observed to cefuroxime (57.7%). MDR S. enterica isolates displayed resistance to ciprofloxacin and azithromycin. Class 1 integrons were detected in 20 (76.9%) of the 26 Salmonella isolates. A high prevalence of class 1 integrons, as the first recorded percentage in the literature, associated with MDR Salmonella isolates was observed. CONCLUSIONS: Antimicrobial resistance rates in Salmonella serotypes from broiler chicken farms were alarming, especially for ciprofloxacin and azithromycin. Thus, another therapeutic strategy other than antimicrobials is recommended to prevent outbreaks of MDR Salmonella.201829684574
2680140.9995Antimicrobial Resistance, Biofilm Formation, and Virulence Genes in Enterococcus Species from Small Backyard Chicken Flocks. Backyard birds are small flocks that are more common in developing countries. They are used for poultry meat and egg production. However, they are also implicated in the maintenance and transmission of several zoonotic diseases, including multidrug-resistant bacteria. Enterococci are one of the most common zoonotic bacteria. They colonize numerous body sites and cause a wide range of serious nosocomial infections in humans. Therefore, the objective of the present study was to investigate the diversity in Enterococcus spp. in healthy birds and to determine the occurrence of multidrug resistance (MDR), multi-locus sequence types, and virulence genes and biofilm formation. From March 2019 to December 2020, cloacal swabs were collected from 15 healthy backyard broiler flocks. A total of 90 enterococci strains were recovered and classified according to the 16S rRNA sequence into Enterococcus faecalis (50%); Enterococcus faecium (33.33%), Enterococcus hirae (13.33%), and Enterococcus avium (3.33%). The isolates exhibited high resistance to tetracycline (55.6%), erythromycin (31.1%), and ampicillin (30%). However, all of the isolates were susceptible to linezolid. Multidrug resistance (MDR) was identified in 30 (33.3%) isolates. The enterococci AMR-associated genes ermB, ermA, tetM, tetL, vanA, cat, and pbp5 were identified in 24 (26.6%), 11 (12.2%), 39 (43.3%), 34 (37.7%), 1 (1.1%), 4 (4.4%), and 23 (25.5%) isolates, respectively. Of the 90 enterococci, 21 (23.3%), 27 (30%), and 36 (40%) isolates showed the presence of cylA, gelE, and agg virulence-associated genes, respectively. Seventy-three (81.1%) isolates exhibited biofilm formation. A statistically significant correlation was obtained for biofilm formation versus the MAR index and MDR. Multi-locus sequence typing (MLST) identified eleven and eight different STs for E. faecalis and E. faecium, respectively. Seven different rep-family plasmid genes (rep1-2, rep3, rep5-6, rep9, and rep11) were detected in the MDR enterococci. Two-thirds (20/30; 66.6%) of the enterococci were positive for one or two rep-families. In conclusion, the results show that healthy backyard chickens could act as a reservoir for MDR and virulent Enterococcus spp. Thus, an effective antimicrobial stewardship program and further studies using a One Health approach are required to investigate the role of backyard chickens as vectors for AMR transmission to humans.202235326843
2138150.9995Isolation and molecular identification of multidrug-resistant Escherichia coli strains isolated from mastitic cows in Egypt. BACKGROUND: Mastitis is a common disease that affects the dairy sector globally because it not only impacts animal welfare but can also lead to significant financial losses. AIM: This study examined the phenotypic and genotypic profiles of the multidrug-resistant (MDR) Escherichia coli (E. coli) strains that were isolated from mastitic cows in Egypt to detect their pattern of antibiotic resistance. METHODS: Four hundred native breed lactating cows were evaluated to identify clinical and subclinical mastitis. A total of 100 mastitic milk samples (64 from clinical mastitis and 36 from subclinical mastitis) were collected for phenotypic isolation and identification of coliform bacteria. Escherichia coli isolates were identified through their morphological features, Gram staining, and biochemical tests. The identified E. coli strains were examined against various antibiotics using disk diffusion methods. All E. coli strains were analyzed for the antibiotic resistance genes Streptomycin (aadA), blaTEM, Tetracycline (tetA), Sulfonamides, and qnrA using PCR. RESULTS: Among 400 examined dairy cows, the prevalences of clinical and subclinical mastitis were 16% and 9%, respectively. Bacteriological isolation of coliform bacteria from mastitic milk samples revealed that E. coli was the most prevalent bacterium. Among 10 isolates of biochemically verified E. coli strains, 8 (80%) were MDR across 6 distinct classes of antibiotics. All recovered E. coli strains exhibited higher resistance to Amoxicillin, Cefotaxime, Sulphamethaxzole/Trimethoprim, and Tetracycline. High susceptibility was noticed to Ciprofloxaccin, Amoxicillin+Clavulinic, Streptomycin, Gentamicin, Chloramphenicol, and Colistin. The blaTEM gene was among the most common antibiotic resistance genes found in E. coli isolates (100%). Furthermore, the genotypes encoding resistance to tetA, aadA, and Sulfonamides were 50%, 40%, and 50%, respectively. CONCLUSION: MDR pathogenic E. coli strains are common in mastitic dairy cows in Egypt, and preventive actions must be implemented to avoid serious public health concerns.202540557079
2154160.9995Molecular analysis of multidrug-resistant E. coli in pediatric UTIs: findings from a Nigerian Hospital. INTRODUCTION: This study aimed to isolate and characterize antibiotic-resistant Escherichia coli from urine samples of children at the Mother and Child Hospital in Ondo State, Nigeria, assessing antibiogram profiling and resistance genes. METHODOLOGY: Three hundred urine samples (158 females, 142 males), aged 3-5 years, were collected, transported on ice, and analyzed bacteriologically. E. coli and Gram-negative bacteria were isolated using Eosin Methylene Blue agar and identified through colony morphology and biochemical tests. Antibiotic susceptibility was determined via Kirby Bauer's disc diffusion, and resistance genes were detected using Polymerase Chain Reaction (PCR). RESULTS: Of the 300 samples, 40 (13.3%) yielded E. coli with varying antibiotic resistance profiles. The highest resistance was against Amoxicillin-clavulanate (87.5%) followed by Ceftriaxone (80%). Susceptibility was observed to Nitrofurantoin, Erythromycin, and Chloramphenicol. Multiple resistance patterns against 3-4 antibiotic classes were recorded, with 12 distinct patterns observed. Eight isolates harbored blaCTX-M gene, while five carried the aac3-IV gene. CONCLUSIONS: The study concluded a high occurrence of E. coli infection and multiple antibiotic resistance in the region. The presence of resistance genes suggests significant economic and health implications, emphasizing prudent antibiotic use under physician guidance to mitigate multiple antibiotic resistance.202438484349
2662170.9995Nasal Carriage of Methicillin-Resistant Staphylococcus Sciuri Group by Residents of an Urban Informal Settlement in Kenya. BACKGROUND: The Staphylococcus sciuri group constitutes animal-associated bacteria but can comprise up to 4% of coagulase-negative staphylococci isolated from human clinical samples. They are reservoirs of resistance genes that are transferable to Staphylococcus aureus but their distribution in communities in sub-Saharan Africa is unknown despite the clinical importance of methicillin-resistant S. aureus. OBJECTIVES: We characterised methicillin-resistant S. sciuri group isolates from nasal swabs of presumably healthy people living in an informal settlement in Nairobi to identify their resistance patterns, and carriage of two methicillin resistance genes. METHOD: Presumptive methicillin-resistant S. sciuri group were isolated from HardyCHROM™ methicillin-resistant S. aureus media. Isolate identification and antibiotic susceptibility testing were done using the VITEK(®)2 Compact. DNA was extracted using the ISOLATE II genomic kit and polymerase chain reaction used to detect mecA and mecC genes. Results: Of 37 presumptive isolates, 43% (16/37) were methicillin-resistant including - S. sciuri (50%; 8/16), S. lentus (31%; 5/16) and S. vitulinus (19%; 3/16). All isolates were susceptible to ciprofloxacin, gentamycin, levofloxacin, moxifloxacin, nitrofurantoin and tigecycline. Resistance was observed to clindamycin (63%), tetracycline (56%), erythromycin (56%), sulfamethoxazole/trimethoprim (25%), daptomycin (19%), rifampicin (13%), doxycycline, linezolid, and vancomycin (each 6%). Most isolates (88%; 14/16) were resistant to at least 2 antibiotic combinations, including methicillin. The mecA and mecC genes were identified in 75% and 50% of isolates, respectively. CONCLUSION: Colonizing S. sciuri group bacteria can carry resistance to methicillin and other therapeutic antibiotics. This highlights their potential to facilitate antimicrobial resistance transmission in community and hospital settings. Surveillance for emerging multidrug resistant strains should be considered in high transmission settings where human-animal interactions are prevalent. Our study scope precluded identifying other molecular determinants for all the observed resistance phenotypes. Larger studies that address the prevalence and risk factors for colonization with S. sciuri group and adopt a one health approach can complement the surveillance efforts.202337529492
1278180.9995Multidrug-resistant enterococci in the hospital environment: detection of novel vancomycin-resistant E. faecium clone ST910. INTRODUCTION: The role of the hospital environment as a reservoir of resistant bacteria in Tunisia has been poorly investigated; however, it could be responsible for the transmission of multidrug-resistant bacteria. The objective was to study the prevalence of Enterococcus in the environment of a Tunisian hospital and the antibiotic resistance phenotype/genotype in recovered isolates, with special reference to vancomycin resistance. METHODOLOGY: A total of 300 samples were taken (March-June, 2013) and inoculated in Slanetz-Bartley agar plates supplemented or not supplemented with 8 µg/mL of vancomycin. Antibiotic resistance genes were tested by polymerase chain reaction (PCR). The clonal relatedness of the vanA isolates was assessed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence testing (MLST). RESULTS: Enterococci were recovered in 33.3% of tested samples inoculated in SB medium. E faecium was the most prevalent species, followed by E. faecalis and E. casseliflavus. Antimicrobial resistance genes detected were as follows (number of isolates): erm(B) (71), tet(M) (18), aph(3')-IIIa (27), ant(6)-Ia (15), cat(A) (4), and van(C2) (6). Vancomycin-resistant-enterococci (VRE) were recovered from 14 samples (4.7%), when tested in SB-VAN. The 14 VRE (one per positive sample) were identified as E. faecium and contained the van(A),erm(B), tet(M), ant(6)-Ia, and aph(3')-IIIa genes. Thirteen of the VRE strains were ascribed by PFGE and MLST to a novel clone (new ST910), and only one VRE strain was typed as ST80 included in CC17. CONCLUSIONS: The emergence and spread of new clones of VRE, especially in the hospital environment in this country, could become particularly problematic.201627580324
1029190.9995Phylogenetic relationships, virulence and antimicrobial resistance properties of Klebsiella sp. isolated from pet turtles in Korea. Klebsiella sp. are responsible for a multitude of infectious diseases in both humans and animals. In this study, phylogenetic relationships, virulence and antimicrobial resistance gene properties of 16 Klebsiella sp. isolated from 49 pet turtles were investigated. The isolates including Klebsiella oxytoca (n = 13) and Klebsiella pneumoniae (n = 3) were identified using 16S rRNA gene sequencing and each species formed distinct clusters in the neighbour-joining phylogenetic tree. The prevalence of virulence genes including ureC (100%) and kfu (68·75%) was observed among the isolates using Polymerase chain reaction (PCR) assay. The fimH, mrkD and rmpA genes were detected in all K. pneumoniae while these were absent in every K. oxytoca isolate. In antimicrobial susceptibility testing, high resistance rates were observed against ampicillin (100%) and cephalothin (62·50%). The resistance rates against imipenem, tetracycline, trimethoprim/sulfamethoxazole, nalidixic acid and ciprofloxacin were 12·50, 12·50, 12·50, 6·25 and 6·25% respectively. The presence of antimicrobial resistance genes such as plasmid-mediated quinolone resistance (PMQR) [qnrB (37·50%), qnrA (31·25%), qnrS (12·50%) and aac(6')-Ib-cr (12·50%)], extended-spectrum β-lactamase (ESBL) [bla(CTX-M) (18·75%)], β-lactamase [bla(SHV-1) (18·75%)] and tetracycline resistance [tetE (12·50%)] was observed. The results revealed that pet turtle-borne Klebsiella sp. may carry different types of virulence and antimicrobial resistance genes which represents a potential threat to public health. SIGNIFICANCE AND IMPACT OF THE STUDY: Klebsiella sp. are nonmotile Gram-negative bacteria that are found in different environments. The virulence and antimicrobial resistance properties of pet turtle-borne Klebsiella sp. have not been studied before. Phylogenetic relationships, virulence traits and antimicrobial resistance profiles of pet turtle-borne Klebsiella sp. were characterized for the first time in Korea. Multiple virulence and antimicrobial resistance genes were observed among the isolates. The occurrence of virulence and antimicrobial resistance determinants in Klebsiella sp. may represent a potential threat to public health.202031671218