Antibiotic resistance profiling and phylotyping of human-diarrheagenic Escherichia coli pathotypes detected from diarrheic and non-diarrheic calves in Iran.

BACKGROUND: Escherichia coli (E. coli) serves as a common indicator of gut microbiota and is utilized for monitoring antimicrobial resistance determinants in food-producing animals. This study aimed to investigate antimicrobial resistance patterns in virulence gene-positive E. coli isolates obtained from 340 healthy and diarrheic calves. METHODS AND RESULTS: A total of 340 fecal swab samples were obtained from diarrheic (n = 170) and healthy (n = 170) calves for 12 months from different farms in Kerman, Iran. The samples were phenotypically analyzed to detect E. coli isolates and antibiotic resistance. Also, antimicrobial resistance genes, diarrheagenic E. coli pathotypes, and phylogenetic background were screened by PCR. Fifteen percent (51/340) of E. coli isolates were positive for at least one of the examined virulence genes (VGs); the prevalence of VGs in E. coli isolates from healthy calves (36/170; 21.17%) was higher than that in diarrheic cases (15/170; 8.82%). Out of the 51 VG-positive isolates, six pathotypes including Shiga toxin-producing E. coli (STEC; 27.45%), enterotoxigenic E. coli (ETEC; 23.52%), enterohemorrhagic E. coli (EHEC; 19.6%), necrotoxigenic E. coli (NTEC; 19.6%), enteropathogenic E. coli (EPEC; 15.68%), enteroinvasive E. coli (EIEC; 1.96%) and three hybrid pathotypes including ETEC/STEC, ETEC/EHEC, and STEC/EIEC were detected among the strains. Antimicrobial resistance (AR) was observed in 98.03% of the VG-positive isolates, which was the same for both healthy and diarrheic calves. The maximum prevalence rate of AR was found against trimethoprim/sulfamethoxazole (49.01%; 3/51), while the minimum prevalence rate was against gentamycin (5.88%; 25/51). Among the VG-positives, phylotype A was found to be the most prevalent followed by B1 and D phylotypes. CONCLUSIONS: The prevalence of VG-positive E. coli isolates was higher in healthy calves compared to diarrheic cases. AR was widespread among VG-positive isolates. These findings suggest that calves may serve as potential reservoirs of antimicrobial-resistant hybrid pathotypes of E. coli.

Explaining the barriers faced by veterinarians against preventing antimicrobial resistance: an innovative interdisciplinary qualitative study.

BACKGROUND: Considering the significance of increased antimicrobial resistance (AMR) and its adverse effects on individual and social health and the important and effective role that veterinarians play in controlling this growing issue worldwide, it is essential to have effective preventive control programs. To this aim, the first step is to identify the factors behind the prevalence of AMR in Iran and the barriers veterinarians face to controlling this problem. Thus, the present study was conducted to explain the barriers veterinarians faced in the prevention of AMR from an Iranian veterinarian's perspective. METHODS: The present research was done in three cities in Iran in 2021. The data were collected through in-depth interviews with 18 veterinarians selected through purposive and snowball sampling and analyzed using conventional qualitative content analysis. RESULTS: The data analysis results were classified into 4 main categories and 44 subcategories. The former included: educational factors, administrative/legal factors, client-related factors, and veterinarian-related factors. CONCLUSIONS: The increased AMR can be approached from multiple aspects. Considering the different factors that affect the increased AMR, it is necessary to consider them all through effective planning and policy-making at multi-level and multidisciplinary dimensions. There is special attention needed to scientific and practical interventions at the individual, interpersonal, social, and even political levels. At the same time, measures should be taken to rehabilitate and maintain the health of society to strengthen supervision and attract the full participation of interested organizations.

An intervention to improve antibiotic prescription behavior in veterinary students: A protocol based on the multi-theory model to tackle antimicrobial resistance.

Background and Aims: Antimicrobial resistance (AMR) is a global health threat. Moreover, incorrect and inappropriate drug prescription behavior is considered a fundamental risk factor. Thus, the present study aims to develop, implement, and evaluate the effectiveness of an educational program based on the multi-theoretical model (MTM) in improving antibiotic prescription behavior in veterinary students of Iran. Methods: The present study will include four phases including a qualitative phase, an instrument design and psychometric test phase, and a cross-sectional, and an interventional phase. In the first phase, the sampling will be purposive with a maximum variety. The interviews will be conducted with a sample of veterinarians. Results: The data will be analyzed in MAXQDA 10. In the second phase, the face and content validity will be tested by a panel of experts as field specialists. A confirmatory factor analysis will be used to test construct validity, and Cronbach's alpha coefficient and intracluster correlation coefficient will be used to determine the internal consistency of the instrument. Then, at this stage, a number of veterinary students will be selected through a multi-stage sampling method. In the cross-sectional phase, another sample of veterinary students will complete a researcher-made questionnaire. Then, Spearman's correlation coefficient test will be used to test the relationship between the two stages of behavior initiation and behavior continuation. The data will be analyzed in SPSS 22. In the third phase, some veterinary students will be selected through a census and will be randomly divided into a control and an intervention group. To collect data in the final phase, the researcher-made questionnaire that was designed in the second phase of the study based on a multi-theory model will be used to extract data. To compare demographic characteristics, compare the correlation between the constructs of the multi-theory model with antibiotic prescribing behavior in the cross-sectional phase and compare the scores of the constructs of the MTM in two intervention and control groups paired-samples T test and independent-samples T test will be used. Conclusion: The present study will aim to improve antibiotic prescription behavior in veterinary students based on a MTM. The findings can be used as a model for training students in clinical fields such as veterinary medicine and general medicine at university at a national level. After verification and approval by experts and university professors, we can expect a change in the educational curriculum to include instructions on how to write out prescriptions for students. There are hopes that the present study if conducted accurately and widely to help prevent AMR in livestock, humans, and society.

Molecular investigation of Coxiella burnetii in aborted fetus of small ruminants in southeast Iran.

The domestic animal, known as a main reservoir of Coxiella burnetii, is susceptible to the occurrence of coxiellosis, which can lead to abortions in domestic animals, causing significant economic damage and posing risks to human health. Therefore, the purpose of this study is to investigate C. burnetii as the causative agent of Q fever in abortion samples of small ruminants in southeastern Iran. This study was conducted between 2020 and 2021 in Zarand city, located in Kerman province (southeast Iran). In this study, 50 abomasum swab samples of aborted sheep and goat fetuses were collected and analyzed using molecular methods to identify C. burnetii. The results revealed that 26% (n: 13) of the collected abortion samples were infected with C. burnetii. Among the positive samples, two (50%) belonged to goat abortion samples while 11 (23.9%) belonged to sheep abortion samples. This study demonstrates that C. burnetii is one of the causes of abortion in small ruminants in southeastern Iran. It is recommended to pay more attention to C. burnetii in domestic animals due to its significant economic impact on livestock and its potential implication for human health in Iran.

Prevalence of canine herpesvirus 1 and associated risk factors among bitches in Iranian breeding kennels and farms.

BACKGROUND: Canine herpesvirus 1 (CHV-1) is an important cause of death in newborn puppies and of fertility problems in adult dogs. Identification of risk factors may help to reduce infection rates and alleviate concerns for dog owners and breeders. This study was designed to screen for CHV-1 infection in bitches of breeding kennels and farms in Iran and relate this to possible risk factors. METHODS: A total of 63 vaginal samples were collected from dogs in 5 breeding kennels (n = 47) and from 7 farms (n = 16). Real-time polymerase chain reaction was used to detect the CHV-1 specific glycoprotein B (gB) gene. Prevalence rates were evaluated in relation to various risk factors, including region, housing, vaccination, deworming, pregnancy, reproductive problems, number of dogs living together and hygiene conditions. RESULTS: In total, 21 (33.3%) of 63 vaginal samples were positive for CHV-1 DNA. The prevalence rate in farms (7/16; 43.7%) was higher than in kennels (14/47; 29.7%). No association was found between CHV-1 prevalence and potential risk factors. CONCLUSIONS: CHV-1 is highly prevalent in dogs in Iranian farms and kennels. Since the CHV1 vaccine is unlicensed in Iran, effective management strategies are essential to reduce the consequences of this pathogen.

High prevalence of ß-lactam and fluoroquinolone resistance in various phylotypes of Escherichia coli isolates from urinary tract infections in Jiroft city, Iran.

BACKGROUND: Urinary tract infection (UTI) is one of the most prevalent infectious diseases with worldwide health threatening. Antimicrobial resistant strains of Escherichia coli (E. coli) are a common cause of UTI which were identified as a treatment challenge. This study aimed to assay the prevalence of common ß-lactam resistance genes including blaTEM, blaSHV, blaCTX-M and blaCMY and phenotypic resistance to commonly used ß-lactam and fluoroquinolone antibiotics in UTIs. These factors were evaluated in various phylogenetic groups (phylotypes) of E. coli isolates. Real-time PCR was applied to detect ß-lactam resistance genes and conventional PCR was used to determine the phylotypes. Phenotypic resistance against ß-lactams (ceftazidime, cefotaxime, aztreonam and ceftriaxone) and fluoroquinolones (ciprofloxacin) were identified by the disc diffusion technique. The ability of extended spectrum ß-lactamases (ESBLs) production in E. coli isolates was detected using the combined disc diffusion method. RESULTS: The prevalence of resistance genes were 89.6% for blaTEM, 44.3% for blaCTX-M, 6.6% for blaSHV and 0.9% for blaCMY. The two high prevalent phylotypes were B2 (29.2%) and D (17.9%) followed by E (14.1%), F (9.4%), C (6.6%) and 10.3% of isolates were unknown in phylotyping. Disc diffusion results showed high prevalence of antibiotic resistance to cefotaxime (88.6%), aztreonam (83%), ceftireaxon (77.3%), ceftazidime (76.4%) and ciprofloxacin (55.6%). Totally, 52.8% of isolates were found as phenotypical ESBL-producers. CONCLUSIONS: This study's results confirmed an explosion of antibiotic resistance amongst E. coli isolates from UTI against ß-lactams and fluoroquinolones. Findings explain the necessity of deep changes in quantity and quality of drug resistance diagnosis and antibiotic therapy strategies. More studies are suggested to better and confident evaluations.

Determinants of antimicrobial resistance occurrence in animal-based food, perceived by livestock farmers: A qualitative phenomenological study.

Background and Purpose of the Study: The determinants of antimicrobial resistance in human medicine have been copiously explored. On the other hand, the existing research in veterinary medicine and animal husbandry is in its infancy. The present qualitative study used the one-health approach to explore farmers' attitudes toward antimicrobial use and antimicrobial stewardship. Methodology: The present qualitative study was phenomenological in type. It was conducted in 2022 in the two cities of Kerman and Bandar Abbas in Iran. The data were collected through in-depth interviews by semistructured interviews with 17 livestock, poultry, and aquatic animal breeders selected through purposive sampling. The interviews lasted between 35 and 65 min in the Farsi language. The data were analyzed using conventional qualitative content analysis and Colaizzi's seven-step analysis technique. Results: The open coding was done in MAXQDA 10 and the results of data analysis were classified into five main themes and 17 subthemes subcategories. The main categories include personal determinants, contextual determinants, legal and regulatory determinants, social determinants, and economic determinants. Conclusion: Considering the increasing use of antibiotics among livestock farmers and breeders of animals used as human food, different measures such as educational, regulatory, social, and even cultural interventions may be able to control and prevent antimicrobial resistance.

Molecular phenotyping of malignant canine mammary tumours: Detection of high-risk group and its relationship with clinicomolecular characteristics.

Canine mammary gland tumours (CMTs) constitute the most common cancer in female dogs and comprise approximately 50% of all canine cancers. With the advent of high-throughput technologies such as microarray and next-generation sequencing, the molecular phenotyping (classification) of various cancers has been extensively developed. The present study used a canine RNA-sequencing dataset, namely GSE119810, to classify 113 malignant CMTs and 64 matched normal samples via an unsupervised hierarchical algorithm with a view to evaluating the association between the resulting subtypes (clusters) (n = 4) and clinical and molecular characteristics. Finally, a molecular classifier was developed, and it detected 1 high-risk molecular subtype in the training dataset (GSE119810) and 2 independent validation datasets (GSE20718 and GSE22516). Our results revealed four molecular subtypes (C2-C5) in malignant CMTs. Furthermore, the normal samples constituted a distinct group in the clustering analysis. Marked significant associations were observed between the molecular subtypes (especially C5) and clinical/molecular features, including positive lymphatic invasion, high tumour grades, histopathology diagnoses, short survival and high TP53 mutation rates (ps <.05). The high-risk subtype (C5) was further characterized through the development of a cell cycle-based gene signature, which comprised 37 proliferation-related genes according to the support vector machine algorithm. This signature identified the high-risk group in both training and validation datasets (ps <.001). In the validation analysis, our potential classifier robustly predicted patients with positive lymphatic invasion, metastases and short survival.

Molecular screening of SARS-CoV-2 in dogs and cats from households with infected owners diagnosed with COVID-19 during Delta and Omicron variant waves in Iran.

OBJECTIVE: The emergence of SARS-CoV-2 infection in dogs and cats in different countries worldwide raises concerns that pets are at a higher risk for spreading or transmitting of SARS-CoV-2 to humans and other pets and increased the research works about the zoonotic aspects and natural routes of infection in companion animals. The current study aimed to detect the SARS-CoV-2 in household dogs and cats living with COVID-19 positive owners. METHODS: Deep oropharyngeal and rectal swabs were collected from 30 household pets (20 cats and 10 dogs) living with COVID-19 positive owners from April 2021 to 2022 in Kerman, Iran. All dogs' and cats' samples were tested by real-time reverse transcription polymerase chain reaction for detection of SARS-CoV-2. RESULTS: Two household cats out of 20 examined (10%) were positive for SARS-CoV-2, whereas none of the examined dogs were positive for SARS-CoV-2. The two cats positive for SARS-CoV-2 were symptomatic and suffered from severe anorexia with maximum contact with their infected owners. CONCLUSION: This study reported the presence of SARS-CoV-2 in household cats in close contact with COVID-19 positive owners during the circulation of new SARS-CoV-2 variants (Delta and Omicron) in Iran and suggested that the transmission may have occurred from owners to their cats. Therefore, infected owners should eagerly limit close contact with their pets during COVID-19 illness.

Highlight of potential impact of new viral genotypes of SARS-CoV-2 on vaccines and anti-viral therapeutics.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causal agent of the coronavirus disease (COVID-19) pandemic, has infected millions of people globally. Genetic variation and selective pressures lead to the accumulation of single nucleotide polymorphism (SNP) within the viral genome that may affect virulence, transmission rate, viral recognition and the efficacy of prophylactic and interventional measures. To address these concerns at the genomic level, we assessed the phylogeny and SNPs of the SARS-CoV-2 mutant population collected to date in Iran in relation to globally reported variants. Phylogenetic analysis of mutant strains revealed the occurrence of the variants known as B.1.1.7 (Alpha), B.1.525 (Eta), and B.1.617 (Delta) that appear to have delineated independently in Iran. SNP analysis of the Iranian sequences revealed that the mutations were predominantly positioned within the S protein-coding region, with most SNPs localizing to the S1 subunit. Seventeen S1-localizing SNPs occurred in the RNA binding domain that interacts with ACE2 of the host cell. Importantly, many of these SNPs are predicted to influence the binding of antibodies and anti-viral therapeutics, indicating that the adaptive host response appears to be imposing a selective pressure that is driving the evolution of the virus in this closed population through enhancing virulence. The SNPs detected within these mutant cohorts are addressed with respect to current prophylactic measures and therapeutic interventions.

High prevalence of clonally related multiple resistant Salmonella Infantis carrying class 1 integrons in broiler farms.

The poultry industry in Iran is the main supplier of protein in the food chain. In the present study, we showed the importance of the possible dissemination of clonally related multiple drug resistant (MDR) Salmonella Infantis in broiler farms in Iran. In total, 156 fecal samples belonging to 23 poultry farms in Razavi Khorasan province, northeast of Iran, were examined for the presence of Salmonella serovars. Molecular serotypes and serogroups, class 1 and 2 integron types, colistin resistance genes ( mcr1 and mcr2) and antimicrobial susceptibility patterns were determined on the recovered Salmonella isolates. Based on PCR analysis, 30 recovered Salmonella isolates were identified asS. Infantis (23 isolates; 76.6%),S. Enteritidis (six isolates; 20%), and one isolate (3.3%) was not serotyped by the applied method. Class 1 integrons were detected in 22 isolates (95.6%) and class 2 integrons were not detected in any of the isolates. Although colistin resistance was prevalent in disc diffusion test, mcr-1 and mcr-2 genes were not detected. All class 1 integrons carried the cassette aadA1 gene. All Salmonella isolates were resistant to colistin and amoxicillin/clavulanic acid and MDR patterns were observed in most (96.6%) isolates. This study revealed a high prevalence rate of S. Infantis and the presence of class 1 integrons in broiler farms. The presence of the same integron cassettes in the sequenced isolates suggests that strains are clonally related. Stringent monitoring programs are required to prevent the spreading of MDR Salmonella serovars into food chain via poultry products.

Serologic and molecular survey of horses to Coxiella burnetii in East of Iran a highly endemic area.

There are few reports about Q fever in horse populations worldwide. This study aimed to detect the C. burnetii infection by serologic and molecular confirmation using commercial ELISA kit and real-time PCR in the East of Iran a region highly endemic. A total of 177 blood samples and 115 vaginal swabs were randomly collected from horses in East of Iran. The sera samples were analyzed for anti C.burnetii Ig G antibodies by a commercial ELISA kit and nucleic acid extraxted from vaginal samples were used to determine the C. burnetii DNA by real-time PCR assay. Antibodies were detected in 5.64 % (10/177) of sera samples and C. burnetii DNA was detected in 7.82 % (9/115) of horse vaginal samples. There was no significant difference in seroprevalence in different sex, age and breed groups. Our study showed that horses could be considered as a mild potential reservoir of C. burnetii which may be effective on horse health status. However, additional studies are needed to assess whether the horse could be considered as a relevant transmission risk indicator for Q fever.

Molecular characterization and antimicrobial resistance of potentially human-pathogenic Escherichia coli strains isolated from riding horses.

BACKGROUND: Transmission of antimicrobial resistant and virulent Escherichia coli (E. coli) from animal to human has been considered as a public health concern. This study aimed to determine the phylogenetic background and prevalence of diarrheagenic E. coli and antimicrobial resistance in healthy riding-horses in Iran. In this research, the genes related to six main pathotypes of E. coli were screened. Also, genotypic and phenotypic antimicrobial resistance against commonly used antibiotics were studied, then phylo-grouping was performed on all the isolates. RESULTS: Out of 65 analyzed isolates, 29.23 % (n = 19) were determined as STEC and 6.15 % (n = 4) as potential EPEC. The most prevalent antimicrobial resistance phenotypes were against amoxicillin/clavulanic acid (46.2 %) and ceftriaxone (38.5 %). blaTEM was the most detected resistance gene (98.4 %) among the isolates and 26.15 % of the E. coli isolates were determined as multi-drug resistant (MDR). Three phylo-types including B1 (76.92 %), A (13.85 %) and D (3.08 %) were detected among the isolates. CONCLUSIONS: Due to the close interaction of horses and humans, these findings would place emphasis on the pathogenic and zoonotic potential of the equine strains and may help to design antimicrobial resistance stewardship programs to control the dissemination of virulent and multi-drug resistant E. coli strains in the community.

Partridge and embryonated partridge egg as new preclinical models for candidiasis.

Candida albicans (C. albicans) is the most common cause of candidiasis in humans and animals. This study was established to a new experimental infection model for systemic candidiasis using partridge and embryonated partridge egg. First, we tested the induction of systemic candidiasis in partridge and embryonated partridge egg. Finally, interaction between virulence factors of C. albicans and Bcl-2 family members was predicted. We observed that embryonic infection causes a decrease in survival time and at later embryonic days (11-12th), embryos showed lesions. Morphometric analysis of the extra-embryonic membrane (EEM) vasculature showed that vascular apoptotic effect of C. albicans was revealed by a significant reduction in capillary area. In immunohistochemistry assay, low expression of Bcl-2 and increased expression of Bax confirmed apoptosis. The gene expression of Bax and Bcl-2 was also altered in fungi-exposed EEM. Ourin silico simulation has shown an accurate interaction between aspartic proteinase, polyamine oxidase, Bcl-2 and BAX. We observed that the disease was associated with adverse consequences, which were similar to human candidiasis. Acquired results support the idea that partridge and embryonated partridge egg can be utilized as appropriate preclinical models to investigate the pathological effects of candidiasis.

First molecular study of Canine herpesvirus-1 in reproductive specimens of adult dogs in southeast of Iran.

Canine herpesvirus-1 (CHV-1) is recognized to be enzootic in the dog population with a widespread distribution. This pathogen leads to a lethal generalized illness in newborn puppies and is associated with reproductive disorders. CHV-1 should be considered as an important pathogen of neonatal death and infertility; so, it appears to pose a threat for breeding kennels. Although serologic data point to the circulation of CHV-1 among dogs of Iran are available, no definitive diagnosis has been conducted based on the molecular assay. So, this research was done to detect the prevalence of CHV-1 in dogs of Kerman. In this study, the presence of CHV-1 in vaginal specimens and biopsies of the uterus of dogs referring to the Veterinary Hospital of Shahid Bahonar University of Kerman was determined. Fifteen uterine samples and seven vaginal samples were included in group of the pregnant dogs. Moreover, thirteen uterine samples and twenty vaginal samples were related to the dogs displaying suspicious clinical signs such as reproductive disorders. Samples were collected and evaluated using real-time PCR. Viral DNA was detected in 21 samples from a total of 140 (15 %) collected samples which were related to 14 uterine samples (20 %) and 7 (10 %) vaginal specimens. The association of this virus with age, breed, housing, pregnancy and reproductive disorders was not significant. Five positive reproductive samples were belonged to the dogs with a history of reproductive disorders including pyometra, metritis, stillbirths, vaginitis and vaginal prolapse. This study is the first molecular detection of CHV-1 in reproductive samples of dogs in Iran. Considering the significant prevalence of this virus, it is necessary to carry out management measures in controlling and preventing this disease. Tracing CHV-1 requires further research on this virus in dogs of this region.

Public health aspects of Shiga toxin-producing Escherichia coli (STEC) strains in sheep and goats of Bakhtiari pastoral tribe, Iran.

Nomadic populations do not have permanent settlements as they move their livestock between grazing areas in different seasons; such movements may have great impact on dissemination of food-borne pathogens in various regions. The aim of this study was to characterize Shiga toxin-producing Escherichia coli (STEC) strains as a food-borne pathogen in sheep and goats of Bakhtiari pastoral tribe in Iran. In the present study, 72 fecal samples were obtained from 26 sheep and 46 goats. First, all recovered E. coli isolates were screened for stx gene. After detection of stx-positive isolates, the virulence genes including stx1, stx2, eae, ehly, saa, astA, subAB, terD, and the genetic markers of O Island 57 (Z2098 and Z2099) were investigated. Also fifteen important STEC O-serogroups were determined using PCR assays. Results showed that 27 animals (27/72; 37.5%) carried STEC strains including 16/26 (61.6%) sheep and 11/46 (23.9%) goats. All STECs were eae-negative but 81.4% (22/27) were positive for saa. The most prevalent virulence profile was stx1/stx2/ehly/saa/subAB (37%; 10/27). Most STECs (24/27) were positive for at least one of the selected OI-57 markers. The O91 (n = 6), O5 (n = 3), O113 (n = 1), O128 (n = 1), and O104 (n = 1) were the detected O-serogroups in this study. It is concluded that such moving animal populations could have public health concerns which have to be addressed in the future.

Domestic and game pigeons as reservoirs for Escherichia coli harbouring antimicrobial resistance genes.

OBJECTIVES: In Iran, pigeons are kept and bred on the roofs of houses, which indicates the potential significance of pigeons in the dispersal of antimicrobial resistant Escherichia coli strains. Here we characterized antimicrobial resistance genotypes in relation to phenotypic presentations and phylogenetic backgrounds of the E. coli isolates from household pigeons in Kerman in southeast Iran. METHODS: Totally, 152 faecal E. coli isolates from domestic and household pigeons were screened for 13 antimicrobial resistance genes, blaTEM, blaSHV, blaCTX-M, sulI, sulII, dhfrI, dhfrV, aadA, aac(3)-I, tetA, tetB, floR and qnrA, by conventional polymerase chain reaction (PCR) technique. Clermont phylogenetic background of E. coli strains was studied and antibiotic resistance of all strains was assessed for seven antibiotics. RESULTS: The antimicrobial resistance genes blaTEM, tetA, tetB and aadA were detected in 52.6%, 6.5%, 6.5% and 5.9% of the isolates, respectively. PCR phylotyping revealed that a significant number of isolates within A0 (54%), A1 (70%), B1 (57.6%), B22 (75%) and D1 (87.5%) phylogroups were positive for the studied resistance genes. One phenotypic resistance pattern (trimethoprim-sulfamethoxazole) was associated with the presence of the corresponding gene sul2. CONCLUSIONS: An alarming rate of phenotypic resistance was observed in this study. Many isolates were positive for the screened resistance genes. According to the phylogenetic background, most resistant isolates belonged to the commensal phylotypes, representing significant role of commensal strains as a source of resistance genes. These findings highlight the role of the pigeon as disseminator of resistant E. coli strains.

Detection of zoonotic diarrheagenic pathotypes of Escherichia coli in healthy household dogs.

BACKGROUND AND OBJECTIVES: Intestinal pathotypes of Escherichia coli belong to the companion animals may poses potential risk to public health following zoonotic transmission. Therefore, this study was proposed to determine the virulence genes associated to diarrheagenic E. coli strains isolated from healthy pet dogs and their owners in the southeast of Iran, Kerman province. MATERIALS AND METHODS: Totally 168 E. coli isolates were collected from 49 healthy household dogs and their owners. Seventy isolates were obtained from non-pet owners as control group. Presence or absence of the virulence genes including eae, stx1, stx2, st1, lt1, ipaH, cnf1 and cnf2 were screened by conventional polymerase chain reaction (PCR) and dissemination pattern of the genes were studied among the various hosts. RESULTS: PCR examinations showed that the most frequent virulence gene was ipaH (6.1%) in dogs followed by eae in dog owners (6.1%) and in controls (8.6%). The most frequent pathotypes in dogs, their owners and controls were EIEC (6.1%), EHEC (4.08%) and EPEC (8.5%), respectively. In one of studied houses, both of dog and its owner harbored E. coli strains with same virulence profile (stx1/eae) and pathotype (EHEC). CONCLUSION: These results collectively indicate that healthy household dogs probably are the mild reservoir of potential virulent E. coli strains with possible active transmission to their contact owner. However, even non-pet owners seemed to be a notable source of intestinal pathotypes, especially EPEC, for their environment. Transmission of E. coli pathotypes may occurs by direct contact with the reservoirs or ingestion of contaminated food. These pathotypes are potentially virulent and creates public health hazards. Further studies are needed for better understanding of dissemination mechanisms of E. coli pathotypes among humans and their pets.

Pathogenic potential of Shiga toxin-producing Escherichia coli strains of caprine origin: virulence genes, Shiga toxin subtypes, phylogenetic background and clonal relatedness.

BACKGROUND: All over the world, Shiga toxin-producing Escherichia coli (STEC) are considered as important zoonotic pathogens. Eight serogroups have the greatest role in the outbreaks and diseases caused by STEC which include O26, O45, O103, O111, O113, O121, O145 and O157. Ruminants, especially cattle are the main reservoirs but the role of small ruminants in the epidemiology of human infections has not been thoroughly assessed in many countries. The objective of this research was to investigate the pathogenic potential of the STEC strains isolated from slaughtered goats. In this study, a total of 57 STEC strains were recovered from 450 goats and characterized by subtyping of stx genes, O-serogrouping, phylo-typing and DNA fingerprinting. RESULTS: Amongst 57 STEC strains isolated from goats, the prevalence of stx1 was significantly more than stx2 (98.2% vs. 24.5%; P ≤ 0.05), and 22.8% of strains harbored both stx1 and stx2 genes. Three (5.2%) isolates were characterized as EHEC, which carried both eae and stx genes. A total of five stx-subtypes were recognized namely: stx1c (94.7%), stx1a (53.7%), stx2d (21%), stx2c (17.5%), and stx2a (15.7%). In some parts of the world, these subtypes have been reported in relation with severe human infections. The stx subtypes predominantly occurred in four combinations, including stx1a/stx1c (35%), stx1c (31.5%), stx1c/stx2a/stx2c/stx2d (5.2%) and stx1c/stx2c/stx2d (%5.2%). In serogrouping, the majority of STECs from goats did not belong to the top 8 serogroups but two strains belonged to O113, which has been recognized as an important pathogenic STEC in Australia. Interestingly, none of stx + eae + isolates belonged to the tested serogroups. In phylo-typing the isolates mostly belonged to phylo-group B1 (82.4%), followed by phylo-group A (12.3%). STEC strains showed a substantial diversity in DNA fingerprinting; there were 24 unique ERIC-types (with a ≥95% similarity) among the isolates. CONCLUSIONS: Despite the fact that the top 8 STEC serogroups were uncommon in caprine strains, the presence of highly pathogenic stx subtypes indicates that small ruminants and their products can be considered as an overlooked public health risk for humans, especially in developing countries which consume traditional products.

Virulence genes, Shiga toxin subtypes, major O-serogroups, and phylogenetic background of Shiga toxin-producing Escherichia coli strains isolated from cattle in Iran.

The aim of this study was to investigate the virulence potential of the isolated bovine STEC for humans in Iran. In this study a collection of STEC strains (n = 50) had been provided via four stages, including sampling from feces of cattle, E. coli isolation, molecular screening of Shiga toxin (stx) genes, and saving the STEC strains from various geographical areas in Iran. The STEC isolates were subjected to stx-subtyping, O-serogrouping, and phylo-grouping by conventional polymerase chain reaction (PCR). Occurrence of stx1 (52%) and stx2 (64%) was not significantly different (p = 0.1), and 16% of isolates carried both stx1 and stx2, simultaneously. In addition, 36% and 80% of the isolates were positive for eae and ehxA, respectively. Molecular subtyping showed that stx1a (52%), stx2a (44%), stx2c (44%), and stx2d (30%) were the most prevalent subtypes; two combinations stx2a/stx2c and stx2c/stx2d coexisted in 18% and 10% of STEC strains, respectively. Three important non-O157 serogroups, including O113 (20%), O26 (12%), and O111 (10%), were predominant, and none of the isolates belonged to O157. Importantly, one O26 isolate carried stx1, stx2, eae and ehxA and revealed highly virulent stx subtypes. Moreover, all the 21 serogrouped strains belonged to the B1 phylo-type. Our study highlights the significance of non-O157 STEC strains carrying highly pathogenic virulence genes in cattle population as the source of this pathogen in Iran. Since non-O157 STEC strains are not routinely tried in most diagnostic laboratories, majority of the STEC-associated human infections appear to be overlooked in the clinical settings.