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This study depicts the drug-resistance and phylogenomic characteristics of 365 Escherichia coli (EC) and 76 Klebsiella pneumoniae (KP) isolated from stray dogs (293) in and around Kolkata, India. Initial screening found 59 isolates, including 48 E. coli and 11 KP multidrug resistant, which included 33 extended-spectrum β-lactamase, 41 AmpC β-lactamase and 18 metallo-β-lactamase producers carrying bla NDM-1 (11) and bla NDM-5 (7) genes. Majority of them had the resistant genes such as bla CTX-M (33), bla TEM (18), bla SHV (4), bla OXA (17), bla FOX (2), bla DHA (2), bla CITM (15), bla CMY-2 (13), bla GES (2) and bla VEB (2), qnr S (15), qnr B (3), aac-6'-Ib-cr (14), tet A (26), tet in phylogroup A (20), B1 (14), and B2 (6). Enterobacteriaceae repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) showed genetic diversity of multidrug resistant isolates irrespective of their origin, resistance, and virulence types, differentiating the EC in five clades (A-E) and KP in four clades (A-D). As these stray dogs, which had no history or scope of previous antimicrobial therapy, were found to have contracted potential antimicrobial resistance pathogens, the role of environment in spread of such pathogens and further possibility of human infections cannot be ruled out.sul -1 (25), arm A (2) and rmt B (6), in addition to adherence genes such as csg A (33), fim A (27), fli C (13), sdi A (33), rcs A (38), and rpo S (39). They also carried plasmid of diverse replicon types of which IncF IA and F IB were the most frequent. Phylogrouping categorized most of the MDR E. coli in phylogroup A (20), B1 (14), and B2 (6). Enterobacteriaceae repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) showed genetic diversity of multidrug resistant isolates irrespective of their origin, resistance, and virulence types, differentiating the EC in five clades (A-E) and KP in four clades (A-D). As these stray dogs, which had no history or scope of previous antimicrobial therapy, were found to have contracted potential antimicrobial resistance pathogens, the role of environment in spread of such pathogens and further possibility of human infections cannot be ruled out.

Objectives: This study aimed to assess the antimicrobial resistance profile, virulence potential, and genetic characterization of avian pathogenic Escherichia coli (APEC) that cause colibacillosis in poultry., Methods: Antibiotic susceptibility testing (AST) was measured via the Kirby-Bauer disc diffusion method against 27 commonly used antibiotics. Phylogrouping, virulence-associated gene detection, and hybrid strain detection via multiplex polymerase chain reaction (PCR) and genetic diversity were analysed via ERIC-PCR fingertyping method., Results: AST analysis showed 100% of isolates were multidrug-resistant (MDR) and highest resistance was against penicillin, tetracycline, and macrolide classes of antibiotics. The mcr-1 gene was present in 40% of the isolates, though only 4% of isolates were showing phenotypic resistance. Despite the scarce use of fluoroquinolone, carbapenem, and cephalosporin in the poultry sector, resistance was evident because of the high prevalence of extended-spectrum β-lactamase (ESBL) (53.7%) and other β-lactamases in APEC isolates. β-lactamase genotyping of APEC isolates revealed that 85.7% of isolates contained either blaCTX or blaTEM and around 38% of isolates were complement resistant. Growth in human urine was evident in 67.3% of isolates. Phylogroup B1 (51%) was the most prevalent group followed by phylogroups A (30.6%), D (13.61%), and B2 (4.76%). The most prevalent virulence-associated genes were fimH, iss, and tatT. Results showed that 26 isolates (17.69%) can be termed hybrid strains and APEC/EHEC (enterohemorrhagic E. coli) was the most prevalent hybrid E. coli pathotype. ERIC-PCR fingerprinting genotype analysis clustered APEC isolates in 40 groups (E1-E40). This study provides insights into the antibiotic resistance and virulence profiling of the APEC isolates in Pakistan., Conclusions: The findings of this study provide insights into that the antibiotic resistance and virulence profiling of the APEC isolates in Pakistan. This data can inform future studies designed to better estimate the severity of the colibacillosis in poultry farms., Competing Interests: Competing interests None declared, (Copyright © 2023. Published by Elsevier Ltd.)

Genotypic diversity among multi-drug-resistant (MDR) aquatic E. coli isolated from different sites of Yamuna River was analyzed using repetitive element PCR (rep-PCR) methods viz. ERIC-PCR and (GTG) 5 -PCR and compared with the MDR animal fecal isolates. The 97 E. coli isolates belonging to different serotypes, phylogroups, and multi-drug resistance patterns were analyzed. High genetic diversity was observed by both the methods; however, (GTG) 5 typing showed higher discriminating potential. Combination of ERIC types (E1-E32) and (GTG) 5 types (G1-G46) generated 77 genotypes. The frequency of genotypes ranged from 0.013 to 0.065. The genotype composition of E. coli isolates was highly diverse at all the sampling sites across Yamuna River except at its entry site in Delhi. The sampling sites under the influence of high anthropogenic activities showed an increase in number of unique genotype isolates. These sites also exhibited high multiple antibiotic resistance (MAR) indexes (above 0.25) suggesting high risk of contamination. Principal coordinate analysis (PCoA) showed limited clustering of genotypes based on the sampling sites. The most frequent genotypes were grouped in the positive zone of both the principal coordinates (PC1 and PC2). The genotypes of most of the animal fecal isolates were unique and occupied a common space in the negative PC1 area forming a separate cluster. High genotypic diversity among the aquatic E. coli and the drain isolates, discharging the untreated municipal waste in the river, was observed, suggesting that the sewage effluents contribute substantially to contamination of this river system than animal feces. The presence of such a high diversity among the MDR E. coli isolates in the natural river systems is of great public health significance and highlights the need of an efficient surveillance system for better management of Indian natural water bodies.

In this study, the prevalence of ESBL/pAmpC-producing Escherichia coli and their molecular characterization from cloacal swab samples were investigated. All samples were obtained from broiler flocks that are located in Hatay, Adana, and Mersin provinces of Turkey. Antimicrobial susceptibilities of the isolates were determined by disk diffusion method following the CLSI criteria. Genetic mechanisms mediating resistance in ESBL/pAmpC-producing E. coli isolates were identified by polymerase chain reaction (PCR) and followed by DNA sequencing. Phylogenetic groups and plasmid replicon types of the isolates were also investigated by PCR. The clonal relationship of selected isolates was investigated by enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus sequence typing (MLST) method. Of 430 cloacal swab samples, 154 (35.8%) were positive for ESBL/pAmpC-producing E. coli. The ESBL/pAmpC type beta-lactamases were as follows: CMY-2 (n = 46), CMY-2 + TEM-1b (n = 63), SHV-12 (n = 5), SHV-12 + TEM-1b (n = 12), CTX-M-3 (n = 14), CTX-M-3 + TEM-1b (n = 1), CTX-M-15 (n = 4), CTX-M-15 + TEM-1b (n = 4), and CTX-M-1 (n = 3). Moreover, various rates of resistance to different antimicrobials were determined such as nalidixic acid (92.9%), ciprofloxacin (76%), sulfamethoxazole-trimethoprim (78.6%), tetracycline (73.4%), streptomycin (52.6%), chloramphenicol (44.2%), kanamycin (27.9%), tobramycin (24.7%), gentamicin (19.5%), and amikacin (0.6%). Furthermore, 148 (96.1%) isolates were found to be MDR. The ESBL/pAmpC-producing isolates were distributed into the following phylogroups: E (n = 61), B1 (n = 30), F (n = 20), A (n = 19), B2 (n = 11), D (n = 10), and C (n = 3). ERIC-PCR analysis showed 51 unrelated patterns. Out of the 28 selected isolates, the following sequence types (STs) were detected: ST354 (n = 3), ST114 (n = 3), ST5696 (n = 2), ST156 (n = 2), ST174 (n = 2), ST362 (n = 2), ST157 (n = 2), ST5114 (n = 2), ST6635, ST539, ST457, ST1640, ST95, ST5843, ST1158, ST10, ST648, and ST4248. The results of the current study revealed that broilers in Turkey are important reservoir of ESBL/pAmpC-producing E. coli, which suggest that these agents have a great potential of transmission to humans by food chain or direct contact.

Aims: In this study, the association between multidrug resistance (MDR) and the expression of some virulence factors were evaluated in Escherichia coli strains isolated from infant faeces and fresh green vegetables. The effect of isolate origin on associated virulence factors was evaluated. In addition, genetic fingerprinting of a sample of these isolates (10 isolates from each group) was studied in order to detect any genetic relatedness among these isolates., Methods and Results: Escherichia coli isolates were divided into four groups based on their origin (human faeces or plant) and their antibiotic resistance (multiresistance or susceptible). PCR was used to investigate heat-labile and heat-stable enterotoxin genes, and four siderophore genes (aerobactin, enterobactin, salmochelin and yersiniabactin). Genetic fingerprinting of the isolates was performed using enterobacterial repetitive intergenic consensus PCR. Siderophore production was measured by a colorimetric method. Biofilm formation was evaluated by a crystal violet assay. The results of the study showed that the expression of MDR is not significantly associated with an increase in these virulence factors or with biofilm formation. However, the origin of isolates had a significant association with siderophore gene availability and consequently on the concentrations of siderophores released. Genetic fingerprinting indicated that human and plant isolates have the same clonal origin, suggesting their circulation among humans and plants., Conclusion: Antibiotic-susceptible strains of E. coli may be as virulent as MDR strains. Results also suggest that the environment can play a potential role in selection of strains with specific virulence factors., Significance and Impact of the Study: Antibiotic-susceptible isolates of Escherichia coli from plant or human origin can be as virulent as the multidrug resistance (MDR) ones. Genetic relatedness was detected among the isolates of plant and human origin, indicating the circulation of these bacteria among human and plants. This could imply a potential role for environmental antimicrobial resistant bacteria in human infection., (© 2019 The Society for Applied Microbiology.)

Background: Urinary tract infections (UTIs) are a highly prevalent infection among children and Escherichia coli is one of the most important pathogens causing pediatric UTIs. Production of extended spectrum b-lactamase (ESBL) enzymes is an important factor in the emergence of antibiotic resistance among these bacteria. This study aimed to determine the resistance patterns, the frequency of ESBL-encoding genes and the genetic diversity of E. coli strains isolated from children with UTIs who were admitted to children's referral hospital of Hazrat Masoumeh, Qom, Iran., Methods: A total of 102 consecutive non-duplicative strains of E.coli that were isolated from children with UTIs were included into the study. Antibiotic susceptibility of the isolates was determined by disk diffusion method according to the CLSI guidelines. The ability of the isolates to produce ESBLs was phenotypically determined by both combined disk test and double disk synergy test. The ESBL encoding genes (bla CTX-M, bla SHV, and bla TEM) in phenotypically confirmed ESBL-positive isolates was detected by PCR method. The genetic relatedness of the isolates was designated by enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR)., Results: Eighty-three percent (n=85) of the children were female. Most of the infected boys (88%, n=15) were less than 1 year of age and most of the infected girls (48%, n=41) aged 1 to 6 years old. The highest sensitivity was observed to nitrofurantoin (8%, n=8), followed by amikacin (12%, n=12) and piperacillin-tazobactam (17%, n=17). In contrast, the highest resistance rate was seen to ampicillin (94%, n=96) and cefazolin (93%, n=95). Eighty-eight percent (90 out of 102) of the strains were multidrug-resistant (MDR). Fifty-eight percent (n=59) of the strains were ESBL-positive and results of the combined disk test was in concordance with PCR. The blaCTX-M was the most frequent ESBL encoding gene (88%, n=52), followed by blaTEM (54%, n=32), and blaSHV (15%, n=9). Based on the ERIC-PCR technique, isolates were clustered in 13 different types. There was no relationship between different ERIC types and origin of the isolates (i.e. hospitalized or outpatients), ESBL-producing ability, and antibiotic resistance patterns., Conclusions: High prevalence of ESBL-positive isolates of E. coli (58%) was found in our study and all of them were MDR. In addition, there were statistically significant differences in the resistance rates of ESBL-producers, and non-producers to some antibiotics, which result in limiting their therapeutic options. Continuous surveillance of the emergence of ESBL-producing isolates and their antibiotic resistance profiles as well as using appropriate typing methods is needed for reducing their spread, selecting appropriate treatment regimens and finding hospital outbreaks.

Objective: The prevalence of Escherichia coli including from ice cubes in Indonesia is quite high. Unfortunately, little is known about the genetic diversity of E. coli from ice cube production site. Genotypic variation in E. coli populations is a major barrier to control public health risk associated with foodborne pathogen. The aims of this study were to analyze the genotypic diversity of E. coli strains isolated from various samples in order to determine the genetic relationship between those strains. This study is also important to understand the occurrence, prevalence and profile picture of different pathogenic E. coli in various sources which potentially cause disease., Results: Enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic polymerase chain reaction (REP-PCR) dendrogram showed high genetic diversity of 120 E. coli isolates in majority of sampling sites. DNA fingerprint patterns showed 26 and 21 clusters with 11 and 3 fingerprints individual lineages for ERIC and REP-PCR respectively. There was no correlation observed between phylogenetic relationship and virulence genes. The result indicated a variation of E. coli isolates in ice cube manufacturers. ERIC-PCR method is more discriminative compared with REP-PCR to analyze the genetic diversity of E. coli from ice cubes production sites.

The extensive use of β-lactam antibiotics has led to emergence and spread of extended-spectrum β-lactamases (ESBLs). This study was conducted to investigate the prevalence of 7 different ESBL genes (bla TEM , bla SHV , bla CTX-M , bla VEB , bla PER , bla GES , and bla OXA-10 ) and O25b-ST131 high-risk clone among 61 clinical isolates of Escherichia coli. Also, one broad-spectrum β-lactamase (bla OXA-1 ) was investigated. This study was also constructed to evaluate iodometric overlay method in detection of ESBL production. Phenotypic identification of E. coli isolates using API 20E revealed 18 distinct biotypes. DNA fingerprinting using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) differentiated all isolates into 2 main phylogenetic groups with 60 distinct genetic profiles. Elevated values of minimal inhibitory concentration (MIC) 50 and MIC 90 for third- and fourth-generation cephalosporins were observed. Phenotypic tests revealed that 85.24% of isolates were ESBL producers. The incidence rates of bla TEM , bla SHV , bla CTX-M , bla GES , bla OXA-1 , and bla OXA-10 among E. coli ESBL producer phenotype were 69.23%, 25%, 96.15%, 3.85%, 11.54%, and 48%, respectively. On the other hand, bla VEB and bla PER were not detected. Sequencing of bla TEM and bla SHV revealed that bla TEM-214 and bla SHV-11 were the most prevalent variants. Group characterization of bla CTX-M revealed that bla CTX-M-1 was the most prevalent group of bla CTX-M family. It was found that 30.77% of E. coli ESBL producers belonged to O25b-ST131 clone harboring bla CTX-M-15 . This study concluded that iodometric overlay method was 100% sensitive in detection of ESBL production. To our knowledge, this is the first Egyptian study that declares the emergence of E. coli O25b-ST131 harboring bla GES .

Unlabelled: Bacterial pathogens producing CTX-M beta-lactamases are emerging around the world as a source of resistance to oxyimino cephalosporins such as cefotaxime. In this study, we have investigated the prevalence of blaCTX-M genes among clinical isolates of Escherichia coli and Klebsiella pneumoniae. Of the double-disk synergy test-positive E. coli (n = 94) and Kl. pneumoniae (n = 73) strains isolated during the study period, 41 (44·08%) E. coli and 32 (43·24%) Kl. pneumoniae isolates were found to be positive for blaCTX-M genes. Twenty-two integrons (13 for E. coli and 9 for Kl. pneumoniae) were detected whose sizes ranged from 600 bp to 1·5 kb. All these integrons were found to be of Class1 type and were invariably PCR positive for int1 and sul1 genes. Marker transfer experiments demonstrated plasmid-mediated transfer of cefotaxime and ceftazidime resistance markers. In addition, analysis of the enterobacterial repetitive intergenic consensus (ERIC)-PCR typing of the blaCTX-M -carrying isolates showed that they were genetically diverse and heterogeneous suggesting that multiple subtypes of the species were involved in infection., Significance and Impact of the Study: A high frequency of blaCTX-M -resistant marker has been found in Escherichia coli and Klebsiella pneumoniae isolates of clinical origin. Analysis of the ERIC-PCR typing of the blaCTX-M -carrying isolates showed that they were genetically diverse and heterogeneous suggesting that multiple subtypes of the species were involved in infection., (© 2016 The Society for Applied Microbiology.)

Captive birds of prey are often used for pest control in urban areas, while also participating in falconry exhibitions. Traveling across the country, these birds may represent a public health concern as they can host pathogenic and zoonotic agents and share the same environment as humans and synanthropic species. In this work, Escherichia coli from the cloacal samples of 27 captive birds of prey were characterized to determine their pathogenic potential. Isolates were clustered through ERIC-PCR fingerprinting, and the phylogenetic groups were assessed using a quadruplex PCR method. Their virulence and resistance profile against nine antibiotics were determined, as well as the isolates' ability to produce extended-spectrum β-lactamases (ESBLs). The 84 original isolates were grouped into 33 clonal types, and it was observed that more than half of the studied isolates belonged to groups D and B2. Most isolates presented gelatinase activity (88%), almost half were able to produce biofilm (45%), and some were able to produce α-hemolysin (18%). The isolates presented high resistance rates towards piperacillin (42%), tetracycline (33%), and doxycycline (30%), and 6% of the isolates were able to produce ESBLs. The results confirm the importance of these birds as reservoirs of virulence and resistance determinants that can be disseminated between wildlife and humans, stressing the need for more studies focusing on these animals. [ABSTRACT FROM AUTHOR]

Background: Uropathogenic Escherichia coli (UPEC) is the main etiological agent behind community-acquired and hospital-acquired urinary tract infections (UTIs), which are among the most prevalent human infections. The management of UPEC infections is becoming increasingly difficult owing to multi-drug resistance, biofilm formation, and the possession of an extensive virulence arsenal. This study aims to characterize UPEC isolates in Tanta, Egypt, with regard to their antimicrobial resistance, phylogenetic profile, biofilm formation, and virulence, as well as the potential associations among these factors. Methods: One hundred UPEC isolates were obtained from UTI patients in Tanta, Egypt. Antimicrobial susceptibility was assessed using the Kirby-Bauer method. Extended-spectrum β-lactamases (ESBLs) production was screened using the double disk synergy test and confirmed with PCR. Biofilm formation was evaluated using the microtiter-plate assay and microscopy-based techniques. The phylogenetic groups of the isolates were determined. The hemolytic activity, motility, siderophore production, and serum resistance of the isolates were also evaluated. The clonal relatedness of the isolates was assessed using ERIC-PCR. Results: Isolates displayed elevated resistance to cephalosporins (90–43%), sulfamethoxazole-trimethoprim (63%), and ciprofloxacin (53%). Ninety percent of the isolates were multidrug-resistant (MDR)/ extensively drug-resistant (XDR) and 67% produced ESBLs. Notably, there was an inverse correlation between biofilm formation and antimicrobial resistance, and 31%, 29%, 32%, and 8% of the isolates were strong, moderate, weak, and non-biofilm producers, respectively. Beta-hemolysis, motility, siderophore production, and serum resistance were detected in 64%, 84%, 65%, and 11% of the isolates, respectively. Siderophore production was correlated to resistance to multiple antibiotics, while hemolysis was more prevalent in susceptible isolates and associated with stronger biofilms. Phylogroups B2 and D predominated, with lower resistance and stronger biofilms in group B2. ERIC-PCR revealed considerable diversity among the isolates. Conclusion: This research highlights the dissemination of resistance in UPEC in Tanta, Egypt. The evident correlation between biofilm and resistance suggests a resistance cost on bacterial cells; and that isolates with lower resistance may rely on biofilms to enhance their survival. This emphasizes the importance of considering biofilm formation ability during the treatment of UPEC infections to avoid therapeutic failure and/or infection recurrence. [ABSTRACT FROM AUTHOR]

Captive birds of prey are often used for pest control in urban areas, while also participating in falconry exhibitions. Traveling across the country, these birds may represent a public health concern as they can host pathogenic and zoonotic agents and share the same environment as humans and synanthropic species. In this work, Escherichia coli from the cloacal samples of 27 captive birds of prey were characterized to determine their pathogenic potential. Isolates were clustered through ERIC-PCR fingerprinting, and the phylogenetic groups were assessed using a quadruplex PCR method. Their virulence and resistance profile against nine antibiotics were determined, as well as the isolates’ ability to produce extended-spectrum β-lactamases (ESBLs). The 84 original isolates were grouped into 33 clonal types, and it was observed that more than half of the studied isolates belonged to groups D and B2. Most isolates presented gelatinase activity (88%), almost half were able to produce biofilm (45%), and some were able to produce α-hemolysin (18%). The isolates presented high resistance rates towards piperacillin (42%), tetracycline (33%), and doxycycline (30%), and 6% of the isolates were able to produce ESBLs. The results confirm the importance of these birds as reservoirs of virulence and resistance determinants that can be disseminated between wildlife and humans, stressing the need for more studies focusing on these animals.

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Abstract There is a global concern and increasing reports regarding foodborne disease infections associated with consuming contaminated vegetables, seafood, meat, and poultry products. Among foodborne bacterial pathogens globally, Salmonella, Escherichia coli, and Shigella were the most frequently implicated in causing food poisoning infections in children and adults. In Saudi Arabia, the consumption rates of imported fresh fruits, vegetables, seafood, and meat products are considered high. Therefore, the development of simple PCR based DNA fingerprinting methods is essential to track the source and route of microbial contamination among imported frozen meat and seafood products. A total of 38 E. coli strains were subtyped using ERIC1R, ERIC2, and a pair combination (ERIC1R + ERIC2) to generate genomic fingerprinting. The three Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR)-based primers were generated in 26, 24, and 16 different genotypes while using ERIC1R, ERIC2, and ERIC1R + ERIC2, respectively. The Discrimination Index values obtained by ERIC1R, ERIC2, and ERIC1R + ERIC2 were 0.976, 0.965, and 0.903, respectively. ERIC1R and ERIC2 primers had the best discriminatory ability and typeability value and proved suitable for investigating genetic analysis among the population of E. coli strains. At the same time, the ERIC1R + ERIC2 primer pair has average discriminatory power and typeability value for differentiating E. coli strains. These results suggest that subtyping using ERIC1R and ERIC2 primer is a more reliable and rapid typing strategy for E. coli strains.

1. The Shiga toxin-producing Escherichia coli (STEC) is a hazardous zoonotic agent for chicken meat consumers. This study determined the serogroups and evaluated the virulence genes, antibiotic resistance, biofilm-forming profiles and genetic relationships of STEC isolates in chicken meat. 2. A total of 100 samples belonging to dressed-whole chicken and different parts of the chicken (wing, breast, thigh, drumstick) were collected between September and November 2019 from different retail markets in Kayseri, Türkiye. 3. Phenotypic (identification, disc diffusion test, Congo red agar and microtitre plate tests) and molecular tests (identification, serogrouping, virulence factors, biofilm, antibiotic susceptibility, 16S rRNA sequencing and enterobacterial repetitive intergenic consensus-PCR for typing of the isolates) were carried out. 4. E. coli was isolated from 35% of the samples and 35% of the samples harboured at least one STEC. Among 35 STEC isolates, 3 (8.5%), 6 (17.1%), 2 (5.7%) and 3 (8.5%) were found to be positive for fliCH2, fliCH8, fliCH11, fliCH19 genes, respectively. Out of 35 STEC positive isolates, 4 (11.4%) were identified as E. coli O157, from which 2 (5.7%) were E. coli O157:H7. E. coli O157 was detected in two (10%), one (5%), one (5%) of the thigh, drumstick and whole chicken samples, respectively. 5. Biofilm-forming ability was reported in 33 (94.2%) of 35 E. coli isolates, whilst the biofilm-associated genes detected among 35 STEC isolates included csgA (88.5%), fimH (88.5%), bcsA (85.7%), agn43 (14.2%) and papC (8.5%). The STEC strains showed resistance against ampicillin (88.5%) and erythromycin (88.5%), followed by tetracycline (74.2%) and gentamicin (25.7%). However, the distribution of isolates harbouring blaCMY, ere(A), tet(A) and aac(3)-IV antibiotic resistance genes was found to be 17.1%, 11.4%, 85.7% and 5.7%, respectively. 6. ERIC-PCR showed that E. coli strains obtained from different parts and whole of chicken samples had genetic diversities. ERIC-PCR patterns grouped strains of 35 STEC into eight clusters designated A-H, with 73% similarity. Proper hygiene measures and staff training are essential for public health during poultry processing and in retail stores to control STEC. [ABSTRACT FROM AUTHOR]

Background: Escherichia coli is a bacterial agent that causes urogenital system infection in dogs. Beta-lactam (ß-lactam) group antibiotics are frequently used in the treatment of E. coli infections. Aims: This study aimed to investigate the presence of extendedspectrum ß-lactamase (ESBL) and plasmidic AmpC in E. coli strains isolated from the urogenital tracts of 125 dogs. Methods: Fifty E. coli strains were identified by conventional bacteriological and PCR methods. Disk diffusion method was used for the determination of antimicrobial susceptibility of the isolates as well as productions of plasmidic AmpC and ESBL. The presence of blaTEM, blaSHV, and blaCTX-M group genes was determined in the isolates by PCR. ERIC-PCR was also used for genotyping of the isolates. Results: Although 22 (44%) of 50 E. coli isolates were found to be ESBL positive, no isolate shows plasmidic AmpC ß-lactamase production. Among 22 ESBL positive isolates, blaTEM, blaSHV, and blaCTX-M group 1 genes were found in 11 (50%), 1 (4.54%), and 6 (27.27%) isolates, respectively. The highest resistance was observed against tetracycline (28%), followed by streptomycin (24%), trimethoprim-sulfamethoxazole (24%), and chloramphenicol (22%), respectively. In the isolates, 11 different main profiles were also determined by ERIC-PCR. It was shown that ESBL positive isolates were related to G10 profiles. Conclusion: The use of extended spectrum ß-lactam group antibiotics for the treatment of E. coli infections in dogs is critical; nevertheless, they may not be effective due to the high rate of resistance to this antibiotic group in E. coli. [ABSTRACT FROM AUTHOR]

There is a global concern and increasing reports regarding foodborne disease infections associated with consuming contaminated vegetables, seafood, meat, and poultry products. Among foodborne bacterial pathogens globally, Salmonella, Escherichia coli, and Shigella were the most frequently implicated in causing food poisoning infections in children and adults. In Saudi Arabia, the consumption rates of imported fresh fruits, vegetables, seafood, and meat products are considered high. Therefore, the development of simple PCR based DNA fingerprinting methods is essential to track the source and route of microbial contamination among imported frozen meat and seafood products. A total of 38 E. coli strains were subtyped using ERIC1R, ERIC2, and a pair combination (ERIC1R + ERIC2) to generate genomic fingerprinting. The three Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR)-based primers were generated in 26, 24, and 16 different genotypes while using ERIC1R, ERIC2, and ERIC1R + ERIC2, respectively. The Discrimination Index values obtained by ERIC1R, ERIC2, and ERIC1R + ERIC2 were 0.976, 0.965, and 0.903, respectively. ERIC1R and ERIC2 primers had the best discriminatory ability and typeability value and proved suitable for investigating genetic analysis among the population of E. coli strains. At the same time, the ERIC1R + ERIC2 primer pair has average discriminatory power and typeability value for differentiating E. coli strains. These results suggest that subtyping using ERIC1R and ERIC2 primer is a more reliable and rapid typing strategy for E. coli strains. [ABSTRACT FROM AUTHOR]

Introduction: The Kingdom of Saudi Arabia has limited data on enteropathogenic Escherichia coli (EPEC). Therefore, this study was undertaken to contribute to EPEC surveillance and investigate the molecular epidemiology of EPEC strains that have been implicated in human infection in King Fahd Hospital of the University (KFHU) between 2013 and 2014 in the Eastern Province of Saudi Arabia. Methods: A total of 60 non-duplicate E. coli isolates associated with human gastroenteritis were included in this study. They were characterized using PCR to determine virulence genes, antimicrobial resistance patterns, and enterobacterial repetitive intergenic consensus (ERIC-PCR). Results: Among the 60 strains, 20% of those examined were positive for the intimin eae and bfpA genes and identified as typical EPEC (tEPEC). Furthermore, 44 of E. coli strains tested positive for the eae gene only and revealed a high occurrence rate of 73.3% of atypical EPEC (aEPEC) within the overall examined strains. All strains were positive for the EAST1 gene, and none tested positive for the stx gene. More than 70% of EPEC strains were multi-drug resistant (MDR) and aEPEC strains with the highest proportion of this feature of MDR. ERIC-PCR fingerprint revealed a total of 19 ERIC types with eight related distinct clusters and a similarity rate cut-off with ≥ 90% homology from the identified isolates. Conclusion: A high antibiotic resistance rate was reported for first-line antibiotics, such as ampicillin, tetracycline, nalidixic acid, trimethoprim-sulfamethoxazole, noroxin, and ciprofloxacin. [ABSTRACT FROM AUTHOR]

The present study aimed to determine the pathogenic Escherichia coli and Salmonella spp. and to investigate their phylogenetic relation by Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) in retail chicken samples. A total of 75 samples were processed for isolation of E. coli and Salmonella spp. by classical cultural methods and isolates were confirmed by the species-specific PCR. Salmonella spp. was detected in 21.3% and E. coli was detected in 74.6% of the chicken carcasses. S. Enteritidis and S. Typhimurium were not detected in chickens by duplex PCR-based assay. O157 based on serotyping and PCR, was not detected in any of the isolates. Besides, virulence and toxin genes were not detected in any of the E. coli isolates. According to ERIC patterns, the obtained ribotypes showed that all Salmonella spp. isolates presented large genetic diversity, whereas only two (3.5%) of E. coli isolates were genetically identical. Although virulent E. coli, and pathogenic serotypes of Salmonella spp. were not detected in our study, it is thought that their high incidence should be considered as an indicator of failure to comply with hygienic conditions and lack of sanitary practices especially in slaughterhouses. [ABSTRACT FROM AUTHOR]

Avian pathogenic Escherichia coli (APEC) and uropathogenic E. coli (UPEC) can cause vast infections in humans and poultry. The present study was conducted to compare the isolates of the APEC and UPEC pathotypes on the basis phenotypic and genotypic features of antibiotic resistance and phylogenetic differences. Total number of 70 identified E. coli strains, including 35 APEC and 35 UPEC isolates, were isolated from avian colibacillosis and human urinary tract infection (UTI), and were subjected to the antimicrobial susceptibility testing, polymerase chain reaction (PCR) detection of the resistance genes, phylogenetic grouping and DNA fingerprinting with enterobacterial repetitive intergenic consensus PCR (ERIC - PCR) to survey the variability of the isolates. The most resistance rates among all E. coli isolates were, respectively, obtained for Ampicillin (84.20%) and sulfamethoxazoletrimethoprim (65.70%). The APEC and UPEC isolates showed the most susceptibility to imipenem and gentamycin, respectively. Among 70 APEC and UPEC isolates 34.20%, 32.80%, 20.00%, and 12.80% belonged to the A, B2, D, and B1 phylogenetic groups, respectively. Analysis of the DNA fingerprinting phylogenetic tree showed 10 specific clusters of APEC and UPEC isolates. According to the results, the most effective antibiotics and the phenotypic and genotypic predominant resistance patterns of the APEC and UPEC isolates were different. Moreover, APECs and UPECs showed various dominant phylogenetic groups. With all descriptions, the APEC isolates still are potential candidates for carrying important resistance genes and can be one of the possible strains related to human infections. [ABSTRACT FROM AUTHOR]

Rapid urbanization has increased human-animal interaction and consequently enhanced the chances to acquire zoonotic diseases. The current investigation is focused to uncover the genetic diversity of multidrug-resistant E. coli strains between different ecologies (i.e., humans, livestock, and environment) at the molecular level by employing antimicrobial resistance profiling, virulence genes profiling, and microbial typing approach using ERIC PCR. Based on multiple antibiotic resistance, overall, 19 antibiotic resistance patterns (R1–R19) were observed. Most of the strains (49/60) were detected to have the combinations of stx, eaeA, and hlyA genes and considered STEC/EPEC/EHEC. A total of 18 unique genetic profiles were identified based on ERIC-PCR fingerprints and most of the strains (13) belong to P1 whereas the least number of strains were showing profiles P7 and P8-P11 (one member each profile). The calculated values for Shannon index (H) for human, animal, and environment are 1.70, 1.82, and 1.78, respectively revealing the highest genetic diversity among the E. coli strains of animal origin. The study revealed that drug-resistant pathogenic E. coli strains could be transmitted bidirectionally among the environment, humans, and animals. [ABSTRACT FROM AUTHOR]

Rapid urbanization has increased human-animal interaction and consequently enhanced the chances to acquire zoonotic diseases. The current investigation is focused to uncover the genetic diversity of multidrug-resistant E. coli strains between different ecologies (i.e., humans, livestock, and environment) at the molecular level by employing antimicrobial resistance profiling, virulence genes profiling, and microbial typing approach using ERIC PCR. Based on multiple antibiotic resistance, overall, 19 antibiotic resistance patterns (R1–R19) were observed. Most of the strains (49/60) were detected to have the combinations of stx, eaeA, and hlyA genes and considered STEC/EPEC/EHEC. A total of 18 unique genetic profiles were identified based on ERIC-PCR fingerprints and most of the strains (13) belong to P1 whereas the least number of strains were showing profiles P7 and P8-P11 (one member each profile). The calculated values for Shannon index (H) for human, animal, and environment are 1.70, 1.82, and 1.78, respectively revealing the highest genetic diversity among the E. coli strains of animal origin. The study revealed that drug-resistant pathogenic E. coli strains could be transmitted bidirectionally among the environment, humans, and animals.

Background and Aim: Rapid and precise typing of E.coli is a prerequisite for epidemiological surveillance and controlling of infection caused by this bacterium. The present study was conducted to determine the molecular diversity of E.coli strains isolated from human and swimming pools samples using Enterobacterial Repetitive Intergenic Consensus (ERIC) method. Materials and Methods: This was descriptive-analytic study was conducted, from 2014 to 2016. The target population in this study were 45 isolates of E.coli (25 isolates from human, 20 isolates from swimming pools sample), all of which were confirmed by biochemical and molecular methods. They were then classified using the ERIC-PCR technique. Results: Generally, in ERIC-PCR product electrophoresis, 3-7 different bands were observed in the range of 100 to 1500 bp. A total of 21 ERIC-PCR profiles were found among all of the studied isolates, that included 12 profiles among human isolates and 10 profiles among swimming pools isolates, and there was one similar profile between them. Based on the results of the dendrogram, 2 main clusters and 7 categories (A to G) were observed. Conclusion: All isolates were typed using ERIC-PCR and high genetic diversity was observed among the isolates. On the other hand, the presence of common profiles among both sources indicates the possible rotation of these isolates between human-water-humans.

One hundred and ten samples of the tap water in Baquba city were collection. Ten isolates were obtained. The results of the sensitivity test showed that 10 isolates were multidrug, 10 isolates exhibited moderate resistance ranging from 5-8 antagonists. Through the use of the RAPD-PCR profiling system, the vast majority of Escherichia coli strains under study have large genomes and high virulence factors by belonging to group A2. Results of genetic profiling using the ERIC-PCR and BOX-PCR system showed several groups of isolates were identified as having a genetic variation and B2 group was given the highest proportion of ERIC-PCR system, 70% in ERIC While A2 group was given the highest proportion of BOX-PCR system, 40% in the BOX indicating that there is a negative spread The present study indicates that there is a correlation between taxonomic systems was evident by the distribution of isolates in each pattern and group. [ABSTRACT FROM AUTHOR]

In this study, it was aimed to investigate the presence of broad-spectrum beta-lactamase (ESBL) producing Escherichia coli in caged birds. A total of 139 fecal swab samples were collected from cage birds from 14 different aviaries in Adana. ESBL producing E. coli was detected in 4.3% (n=6) of the fecal samples and these isolates harbored blaCTX-M-15 gene. ERIC-PCR analysis revealed three different band patterns among the isolates. These results indicate that cage birds are carrier of ESBL-producing E. coli. Therefore, further epidemiological studies are needed to determine the presence of resistant bacteria including ESBL producing E. coli in various animal species.

The aim of this study was to detect virulence genes and drug resistance genes in pathogenic Escherichia coli (E.coli) isolated from diarrhea calves feces and to detect drug resistance phenotypes, and genotypic. As a result, a total of 22 strains of E.coli were separated and identified, but only 18 strains were pathogenic. The virulence gene test showed that the LT, Stx1, F5, F6, and irp2 gene detection rate were 66.67% (12/18), 16.67% (3/18), 55.56% (10/18), 44.44% (8/18) and 16.67% (3/18), respectively. Antibiotic susceptibility tests showed that the isolates were susceptible to Kanamycin and Florfenicol and resistant to Cefalotin (94.44%) and the like. Resistance gene tet (A), gryA, parC, sulI, sulII, sulIII, blaTEM, blaCTX-m, blaSHV, aac (3)-II, aac (6 ')-Ib and rmtB detection rate are 77.78% (14/18), 44.44% (8/18), 55.56% (10/18), 22.22% (4/18), 11.11% (2/18), 11.11% (2/18), 77.78 % (14/18), 5.56% (1/18), 44.44% (8/18), 50% (9/18), 55.56% (10/18), and 27.78% (5/18), respectively. ERIC-PCR (repetitive intergenic consensus-polymerase chain reaction) classified 18 pathogenic E.coli into 4 distinct genotypes, and all strains carrying the LT virulence gene were brought together, but there was no link between drug resistance genes and genotypic. The results of this study provids experimental evidence for the prevention, control pathogenic E. coli disease in calves. [ABSTRACT FROM AUTHOR]

The drug resistance of Escherichia coli from animals was detected in Shandong Province in 2016, the gene mapping of Enterobacterial Repetitive Intergenic Consensus-PCR(ERIC-PCR) of multidrug-resistant isolates were analyzed, and then the relationship between ERIC-PCR genotyping and drug resistance of highly-resistant E. coli with multidrug resistance was discussed. A total of 110 E. coli isolates were separated and identified from diseased swine and avian, and their resistance to 10 kinds of antimicrobials was determined by the agar dilution method. Twenty highly-resistant isolates with multidrug resistance were selected to carry on the ERIC-PCR, followed by cluster genetic analysis according to the DNA fingerprints. The swine-sourced E. coli isolates possessed serious resistance against several kinds of antimicrobial agents, for example, all isolates were tolerant to florfenicol, doxycycline and ampicillin(100%), but had a relative lower resistance rate to cefotaxime sodium(57.14%). The same situation was observed in the poultry-sourced E. coli isolates, which had a resistance rate of 95.51% against florfenicol, while the lowest rate of 61.80% appeared on ciprofloxacin. Analysis of ERTIC showed dispersive fingerprint patterns of highly-resistant and multidrug-resistant isolates which represented a multiple clone resources, and there were certain correlation between the B genotype and the drug-resistant characteristics. It indicated that the animal-sourced E. coli isolates had a high level of drug resistance and were multidrug-resistant, which meant there was severe antibiotic resistance against not only different kinds of antibiotics but also different drugs of the same kind. The highly-resistant E. coli isolates with multidrug resistance had no apparent species preference, while their spread and pervasion posed a potential threat to the development of animal husbandry and the public health security. [ABSTRACT FROM AUTHOR]

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Shiga toxin‐producing Escherichia coli strains (STEC) are food‐borne pathogens. While E. coli O157:H7 is commonly associated with cattle, less is known about the prevalence of non‐O157 STEC serogroups in bovines. This study evaluated the prevalence and virulence status of O157:H7 and six E. coli O‐serogroups (O26, O103, O45, O145, O121, O111) in New Zealand dairy farms using molecular as well as culture‐based methods. Fresh farm dairy effluent (FDE) (n = 36) and composite calf faeces (n = 12) were collected over three samplings from 12 dairy farms. All seven target serogroups were detected through molecular techniques. Of the 202 isolates which were serologically confirmed following traditional culturing and immunomagnetic separation (IMS), O103, O26, O45 and O121 were the most common serogroups, being found in 81, 47, 42 and 32% of the FDE and in 17, 33, 25 and 9% of the calf faeces respectively. The majority (157/202) of the isolates were negative for stx and eae virulence genes. The prevalence of the seven target STEC was low, and only nine O26 isolates (4%) were recovered from four of the farms. The study has highlighted the need for improving the isolation of Top 7 STEC from the stx‐negative populations present in fresh dairy effluent and calf faeces. Significance and Impact of the Study: Shiga toxin‐producing Escherichia coli (STEC) are important food‐borne pathogens that can cause severe illness in humans. Cattle are asymptomatic reservoirs for STEC, and transmission to humans can be by consumption of food products or water contaminated with cattle faeces. Our study investigated the prevalence of O157:H7 and six E. coli serogroups of STEC (O26, O103, O45, O145, O121, O111) over time in the dairy reservoir and increases the knowledge and understanding of these pathogens on pasture‐based farms. Such information is required to develop risk‐assessment models aiming at limiting transmission of these STEC to human. Significance and Impact of the Study: Shiga toxin‐producing Escherichia coli (STEC) are important food‐borne pathogens that can cause severe illness in humans. Cattle are asymptomatic reservoirs for STEC, and transmission to humans can be by consumption of food products or water contaminated with cattle faeces. Our study investigated the prevalence of O157:H7 and six E. coli serogroups of STEC (O26, O103, O45, O145, O121, O111) over time in the dairy reservoir and increases the knowledge and understanding of these pathogens on pasture‐based farms. Such information is required to develop risk‐assessment models aiming at limiting transmission of these STEC to human. [ABSTRACT FROM AUTHOR]

Background and Aim: Rapid and precise typing of E.coli is a prerequisite for epidemiological surveillance and controlling of infection caused by this bacterium. The present study was conducted to determine the molecular diversity of E.coli strains isolated from human and swimming pools samples using Enterobacterial Repetitive Intergenic Consensus (ERIC) method. Materials and Methods: This was descriptive-analytic study was conducted, from 2014 to 2016. The target population in this study were 45 isolates of E.coli (25 isolates from human, 20 isolates from swimming pools sample), all of which were confirmed by biochemical and molecular methods. They were then classified using the ERIC-PCR technique. Results: Generally, in ERIC-PCR product electrophoresis, 3-7 different bands were observed in the range of 100 to 1500 bp. A total of 21 ERIC-PCR profiles were found among all of the studied isolates, that included 12 profiles among human isolates and 10 profiles among swimming pools isolates, and there was one similar profile between them. Based on the results of the dendrogram, 2 main clusters and 7 categories (A to G) were observed. Conclusion: All isolates were typed using ERIC-PCR and high genetic diversity was observed among the isolates. On the other hand, the presence of common profiles among both sources indicates the possible rotation of these isolates between human-water-humans. [ABSTRACT FROM AUTHOR]

Tempeh is a famous Indonesian fermented food derived from soybeans inoculated with Rhizopus sp. Tempeh production varies depend on the producers and often conducted in an uncontrolled condition. This condition could lead to the growth of Escherichia coli which is known as bacterial indicators of environmental hygiene. Some strains of E. coli could induce diarrhea, acute gastroenteritis or gastrointestinal tract infections. The aim of this study was to compare genetic diversity of E. coli isolates from tempeh with medical isolates employing ERIC-PCR method. In this study, 63f and 1387r primers were used to amplify 16S rRNA genes, and ERIC 1R and ERIC 2 primers were used for ERIC-PCR analysis. Tempeh samples were obtained from four producers in Bogor. Thirty-three isolates of E. coli were successfully isolated from tempeh samples produced by only two producers, we could not obtain E. coli isolates from the other two producers. In addition, the same tempeh samples could carry different genotypes of E. coli. On the other hand, the same genotypes could be found in different tempeh samples. Based on phylogenetic tree analysis, E. coli from tempeh could be separated from medical isolates. We showed that E. coli isolates derived from tempeh were genetically different from those of medical or pathogenic isolates.

In this study, it was aimed to investigate the presence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli in caged birds. A total of 139 fecal swab samples were collected from cage birds from 14 different aviaries in Adana. ESBL producing E. coli was detected in 4.3% (n=6) of the fecal samples and these isolates harbored blaCTX-M-15 gene. ERIC-PCR analysis revealed three different band patterns among the isolates. These results indicate that cage birds are carrier of ESBL-producing E. coli. Therefore, further epidemiological studies are needed to determine the presence of resistant bacteria including ESBL producing E. coli in various animal species. [ABSTRACT FROM AUTHOR]

The extensive use of β-lactam antibiotics has led to emergence and spread of extended-spectrum β-lactamases (ESBLs). This study was conducted to investigate the prevalence of 7 different ESBL genes (blaTEM, blaSHV, blaCTX-M, blaVEB, blaPER, blaGES, and blaOXA-10) and O25b-ST131 high-risk clone among 61 clinical isolates of Escherichia coli. Also, one broad-spectrum β-lactamase (blaOXA-1) was investigated. This study was also constructed to evaluate iodometric overlay method in detection of ESBL production. Phenotypic identification of E. coli isolates using API 20E revealed 18 distinct biotypes. DNA fingerprinting using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) differentiated all isolates into 2 main phylogenetic groups with 60 distinct genetic profiles. Elevated values of minimal inhibitory concentration (MIC)50 and MIC90 for third- and fourth-generation cephalosporins were observed. Phenotypic tests revealed that 85.24% of isolates were ESBL producers. The incidence rates of blaTEM, blaSHV, blaCTX-M, blaGES, blaOXA-1, and blaOXA-10 among E. coli ESBL producer phenotype were 69.23%, 25%, 96.15%, 3.85%, 11.54%, and 48%, respectively. On the other hand, blaVEB and blaPER were not detected. Sequencing of blaTEM and blaSHV revealed that blaTEM-214 and blaSHV-11 were the most prevalent variants. Group characterization of blaCTX-M revealed that blaCTX-M-1 was the most prevalent group of blaCTX-M family. It was found that 30.77% of E. coli ESBL producers belonged to O25b- ST131 clone harboring blaCTX-M-15. This study concluded that iodometric overlay method was 100% sensitive in detection of ESBL production. To our knowledge, this is the first Egyptian study that declares the emergence of E. coli O25b-ST131 harboring blaGES. [ABSTRACT FROM AUTHOR]

In this study, genotypic and phenotypic characteristics of the strains, antibiotic susceptibility, and antibiotic resistance genes of the Escherichia coli, Bacillus cereus, Listeria spp., and Salmonella spp. isolated from various animal foods in Sinop province were determined. Resistance percentages of the isolated strains to the antibiotics penicillin, vancomycin, clindamycin, tetracycline, ampicillin/sulbactam, gentamicin, ciprofloxacin, and chloramphenicol were 87.8, 71.9, 69.7, 63.4, 27.5, 23.1, 20.9, and 13.4%, respectively. Antibiotic resistance genes tetA, blaCMY-2, and cat1 were found in 45, 21.25, and 6.25% of all the strains, respectively. The virulence genes were also investigated in the isolated in E. coli strains; the eae gene was found in 13.1% of the strains, while the stx1 was not identified in any strains. As a result, we can say that SDS-PAGE method may provide discrimination at species level; however, surveys at subspecies level demand molecular approaches such as ERIC and (GTG)5-PCR. In addition, we can suggest that molecular techniques may be especially helpful in the rapid identification of multidrug-resistant and isolates with virulence genes. This study has revealed that the investigated bacteria in animal foods is a significant reservoir of resistance genes. [ABSTRACT FROM AUTHOR]

Shiga toxin producing (STEC), enteropathogenic (EPEC) and enterotoxigenic Escherichia coli (ETEC) were isolated from 29 diarrhoeic yak calves from two districts of Arunachal Pradesh, India (West Kameng and Tawang) during 2005 to 2011. The STEC (28) and EPEC (12) isolates belonged to 25 different O serogroups. Among the 28 STEC isolates, 8 (28.5%) isolates carried only stx1, 10 (35.7%) isolates were positive for both stx1 and stx2, 10 (35.7%) isolates carried only stx2. The stx variants such as stx1c, stx2c, stx2d and stx2e were detected in 7 (25%), 9 (32.1%), 1 (3.5%) and 4 (14.28%) isolates, respectively. Among the 12 EPEC isolates, 2 (4.65%) strains were ‘typical’ possessing bfpA gene. The ehxA and saa genes were present in 19 and 4 isolates, respectively. Among the 14 ETEC isolates, 6 (42.8%), 7 (50%), 5 (35.7%), 5 (35.7%), 7 (50%) and 6 (42.8%) isolates carried genes coding for STa, STb, LT, F5, F41 and EAST1, respectively. The present study detected LT, STb, F41 as the most prevalent type in yaks associated with diarrhoea. Further, the STEC, EPEC and ETEC isolates showed resistance against furazolidone (40%), nitrofurantoin (39%), nalidixic acid (38%), erythromycin (38%), kanamycin (38%), amikacin (37%). The dendrogram revealed clonal relationship among the isolates after RAPD/ ERIC analysis. The study could not detect any specific RAPD/ERIC cluster associated with origin, serotypes or virotypes of the isolates. Thus, it was concluded that genetically diverse, multi-drug resistant enterovirulent E. coli were associated with death due to diarrhoea in yak calves in India. [ABSTRACT FROM AUTHOR]

The prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) is increasing rapidly in both hospital environments and animal farms. A lot of animal manure has been directly applied into arable fields in developing countries. But the impact of ESBL-positive bacteria from animal manure on the agricultural fields is sparse, especially in the rural regions of Tai'an, China. Here, we collected 29, 3, and 10 ESBL-producing E. coli from pig manure, compost, and soil samples, respectively. To track ESBL-harboring E. coli from agricultural soil, these isolates of different sources were analyzed with regard to antibiotic resistance profiles, ESBL genes, plasmid replicons, and enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR) typing. The results showed that all the isolates exhibited multi-drug resistant (MDR). CTX-M gene was the predominant ESBL gene in the isolates from pig farm samples (30/32, 93.8%) and soil samples (7/10, 70.0%), but no SHV gene was detected. Twenty-five isolates contained the IncF-type replicon of plasmid, including 18 strains (18/32, 56.3%) from the pig farm and 7 (7/10, 70.0%) from the soil samples. ERIC-PCR demonstrated that 3 isolates from soil had above 90% genetic similarity with strains from pig farm samples. In conclusion, application of animal manure carrying drug-resistant bacteria on agricultural fields is a likely contributor to antibiotic resistance gene spread. [ABSTRACT FROM AUTHOR]

Escherichia coli and Escherichia coli O157 were identified from "selom" (Oenanthe stolonifera), "pegaga" (Centella asiatica), beef, chicken, lamb, buffalo, "ulam Raja" (Cosmos caudatus) and "tenggek burung" (Euodia redlevi). The bacteria were recovered using chromagenic agar. Isolated Escherichia coli and Escherichia coli 0157 were further characterized by plasmid profiling and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The virulence genes of the isolates (VT1, VT2, LT, ST, eaeA, inV) that produces pathogenic Escherichia coli and 16S rRNA gene were screened by a multiplex PCR assay. The plasmid profiling analysis showed that out of 176 isolates, only 103 isolates contained plasmids. ERIC-PCR analysis generated amplified products in the range of ~150 bp to > 1000 bp categorizing isolates into a total of 52 different profiles. Multiplex PCR showed that 20 (32.3%) of the isolates carried eaeA gene, 6 (9.7%) isolates possessed inV genes, only 1 (1.6%) have VT2 genes and 1 (1.6%) as well carried VT1 genes, 2 (3.2%) of the isolates harboured LT genes, and only 1 (1.6%) isolate possessed ST genes. There were no correlation between plasmid, ERIC-PCR and virulence genes profiles. [ABSTRACT FROM AUTHOR]

Abstract: To understand the spread of microbial aerosols in pig houses, with Escherichia coli (E. coli) as indicator, the airborne E. coli in 4 pig houses and their surroundings at different points 10, 50m upwind and 10, 50, 100, 200 and 400m downwind respectively from the pig houses were collected, and the concentrations were calculated at each sampling point. Furthermore, the feces of pigs were collected to separate E. coli. The ERIC-PCR (Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction) technology was used to amplify the isolated E. coli DNA samples, then the amplified results were analyzed by NTSYS-pc (Version 2.10) to identify the similarity of isolated E. coli. The results showed that the airborne E. coli concentrations in indoor air of the 4 pig houses (21–35CFUm− 3 air) were much higher than those in upwind and downwind air (P <0.05), but there were no significant differences (P >0.05) at downwind distances. The ERIC-PCR results also showed that 52.4% of the fecal E. coli (four houses being respectively 2/4, 50%; 2/4, 50%; 3/6, 50%; 4/7, 57.1%) were identical to the indoor airborne E. coli isolates, and there was more than 90% similarity between the majority of E. coli (50%, 21/42) isolated from downwind air at 10, 50, 100 and 200m and those from indoor air or feces. It could be concluded that the aerosols in pig houses can spread to the surroundings, and thus effective measures should be taken to control and minimize the spread of microbial aerosols. [Copyright &y& Elsevier]

Aims: Fresh fruits and vegetables are increasingly recognized as vectors for food-borne illness. On farm contamination through contaminated irrigation water is considered likely source of the pathogen for several outbreaks. The purpose of this study is to investigate the possible similarity of strains of Escherichia coli isolated from the soil and vegetables irrigated by treated wastewater. Methods and Results: Seventy-five strains of enteropathogenic Escherichia coli isolated from vegetables, soil and irrigation water were tested for sensitivity to antibiotics and shown to be sensitive. The result of enterobacterial repetitive intergenic consensus (ERIC)-PCR shows similarities between analysed strains isolated from the three different samples. Moreover strains of E. coli isolated from vegetables over different periods of time have the same ERIC-PCR profile. Conclusions: The isolated strains of enteropathogenic E. coli can persist in soil and in vegetables growing in fields treated with contaminated irrigation water for an extended period of time. Significance and Impact of the Study: Contaminated irrigation water can transport pathogenic bacteria, which persists in the soil for a long period of time and contaminates the vegetables growing in the field irrigated by this contaminated water. [ABSTRACT FROM AUTHOR]

Among the 51 clinical isolates collected from a university hospital in Korea, nine isolates were resistant to cephamycins. Nine isolates were shown to produce CMY-11 and these also included three isolates producing TEM-1. The results from ERIC-PCR revealed that dissemination of CMY-11 was due to outbreaks of resistant species and to the intra-species spread of resistance to cephamycins in Korea. CMY-11 β-lactamase genes from nine clinical isolates that were responsible for resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin and amoxicillin-clavulanic acid, were cloned and characterised. A sequence identical to the common regions in In6, In7 and a novel integron from pSAL-1 was found upstream from blaCMY-11 gene at nucleotide 1–71. Eighteen nucleotides between position 71 and 72 were inserted into the blaCMY-11 gene. [Copyright &y& Elsevier]

Forty-nine avian Escherichia coli isolates isolated from different outbreak cases of septicemia (24 isolates), swollen head syndrome (14 isolates) and omphalitis (11 isolates), and 30 commensal isolates isolated from poultry with no signs of illness were characterized by enterobacterial repetitive intergenic consensus (ERIC)–PCR technique and their serotypes were determined. The ERIC–PCR profile allowed the typing of the 79 isolates into 68 ERIC-types and grouped the isolates into four main clusters (A–D), with the omphalitis isolates being grouped with the commensals and separated from the septicaemia and swollen head syndrome. These results indicate that ERIC–PCR is a technique that could replace other molecular characterization techniques such as random amplification of polymorphic DNA (RAPD)–PCR and restriction fragment length polymorphism (RFLP), reinforce previous observations that omphalitis isolates are just opportunistic agents, and are consistent with many reports that specific genotypes are responsible for causing specific diseases. Most of the isolates were either nontypable or rough, supporting the need for alternative methods for typing these isolates. [Copyright &y& Elsevier]