Virulence Genes Stx1 Research Articles

Escherichia coli and Enterobacteriaceae Counts, Virulence Gene Profile, Antimicrobial Resistance, and Biofilm Formation Capacity during Pig Slaughter Stages.

This study aimed to count Enterobacteriaceae and Escherichia coli in different locations on pig carcasses (shank, loin, abdomen, shoulder, and jowl) from two slaughterhouses (A and B) between September 2019 and July 2021 during different slaughter stages (after bleeding, after passing through the epilator machine, after manual toileting in the dirty area, before and after evisceration, and after the final washing), as well as verify antimicrobial resistance and biofilm formation capacity. The main points of Enterobacteriaceae and E. coli contamination were identified in the two slaughterhouses through three collections. The stages with the highest counts were post-bleeding and evisceration in both slaughterhouses and after manual toileting in slaughterhouse B in the first collection. Most E. coli isolates were resistant to multiple antimicrobials, with higher resistance frequencies to amoxicillin, ampicillin, chloramphenicol, sulfonamides, and streptomycin. The virulence genes eae, stx1, and stx2 were also detected. Three isolates had all three genes and exhibited resistance to at least six antimicrobial classes (β-lactams, macrolides, aminoglycosides, sulfonamides, amphenicols, and quinolones). E. coli isolates also showed a high frequency of strains with moderate and strong in vitro biofilm-forming capacity. This is the first study to characterize microbial contamination by pig slaughter stage in the Federal District region, demonstrating the critical points for hygienic production. E. coli was isolated from the surface of pig carcasses, as well as the virulence genes stx1, stx2, and eae were detected. The multi-antimicrobial resistant isolates also had a moderate-to-strong biofilm formation capacity, thus demonstrating risks to public health.

Differential Age-Based Response Induced by a Commercial Probiotic Supplementation in Pastured Goats.

The potential benefit of probiotics in small ruminant production systems has largely been unexplored. We evaluated the effect of a goat commercial probiotic on health and performance indicators in pastured goats from birth until 10months. We randomly allocated 26 newborn nursing goat kids to two groups: a control group that received saline and a treatment group that received a commercial probiotic paste orally. We evaluated select observable health indicators (inappetence, diarrhea, coughing), weight, immunity (IgA, IgG, and innate immune response), total protein, hematocrit (HCT), total lactic acid bacteria (LAB), total coliforms, and prevalence of Escherichia coli (E. coli) primary virulence genes (stx1, stx2, and eae) during the experimental period. The results revealed no significant differences in the health indicators, LAB count, and total E. coli count. Prevalence of stx1 at 1week of age and both stx1 and stx2 genes 4months post-weaning was significantly (P < 0.05) higher in probiotic-supplemented goats. Probiotic supplementation significantly (P < 0.05) increased the total protein and IgA 1month post-supplementation during the pre-weaning period and innate immune markers 2days post-weaning. The HCT in probiotic-supplemented goats was significantly (P < 0.05) higher at 1 and 2months post-weaning. The growth rate was not affected by probiotic supplementation in pre- and peri-weaned goats but was significantly (P < 0.05) lowered in goats older than 4months in the supplemented group. In this pastured goat production study, there were mixed responses to a commercial probiotic in healthy goats based on age. The study suggests that early daily probiotic supplementation in pre-weaned pastured goats may have immune stimulation benefits, but in older healthy animals, post-weaning net benefits are unclear and further research is recommended.

Occurrence, antibiotic resistance and molecular characterisation of Shiga toxin-producing Escherichia coli isolated from broiler chickens in Tunisia

ABSTRACT 1. Shiga toxin-producing Escherichia coli (STEC) strains are associated with disease outbreaks which cause a public health problem. The aim of this study was to determine the frequency of STEC strains, their virulence factors, phylogenetic groups and antimicrobial resistance profiles in broiler chickens. 2. A total of 222 E.coli isolates were collected from the caecum of chickens intended to be slaughtered. Antibiotic susceptibility was tested against 21 antimicrobial agents and ESBL phenotype was assessed by double-disk synergy test. The presence of STEC virulence genes stx1, stx2,eaeA and ehxA was detected by PCR. The identification of STEC serogroups was realised by PCR amplification. Additive virulence genes, phylogenetic groups and integrons were examined among the STEC isolates. 3. Out of 222 E.coli isolates, 72 (32%) were identified as STEC strains and the most predominant serogroups were O103, O145 and O157. Shiga toxin gene 1 (stx1) was found in 84.7% (61/72) of the STEC strains, and eae and stx2 were detected in 38.8% and 13.8%, respectively. The ESBL phenotype was documented in 48.6% (35/72) of isolates. Most of the isolates (90.3%) carried class 1 integron with the gene cassette encoding resistance to trimethoprim (dfrA) and streptomycin (aadA) in 31.9% of the isolates. Class 2 integron was identified in 36.1% of isolates. 4. Broilers can be considered as a reservoir of STEC strains which have high virulence factors and integrons that might be transmitted to other chickens, environments and humans. It is important to undertake surveillance and efficient control measures in slaughterhouses and farms to control measures of STEC bacteria.

Multiple-Drug Resistant Shiga Toxin-Producing E. coli in Raw Milk of Dairy Bovine.

Raw milk may contain pathogenic microorganisms harmful to humans, e.g., multidrug-resistant Escherichia coli non-O157:H7, which can cause severe colitis, hemolytic uremia, and meningitis in children. No studies are available on the prevalence of Shiga toxin-producing E. coli (STEC O157:H7) in sick or healthy dairy animals in the Khyber Pakhtunkhwa Province of Pakistan. This study aimed to isolate, characterize, and detect antibiotic resistance in STEC non-O157:H7 from unpasteurized milk of dairy bovines in this province. We collected raw milk samples (n = 800) from dairy farms, street vendors, and milk shops from different parts of the Khyber Pakhtunkhwa Province. E. coli was isolated from these samples followed by latex agglutination tests for serotyping. The detection of STEC was conducted phenotypically and confirmed by the detection of virulence genes genotypically. An antibiogram of STEC isolates was performed against 12 antibiotics using the disc diffusion method. A total of 321 (40.12%) samples were found to be positive for E. coli in this study. These samples were processed for the presence of four virulence genes (Stx1, Stx2, ehxA, eae). Forty samples (5.0%) were STEC-positive. Of these, 38%, 25%, 19%, and 18% were positive for Stx1, Stx2, ehxA, and eae, respectively. Genotypically, we found that 1.37% of STEC isolates produced extended-spectrum beta-lactamase (ESBL) and contained the blaCTX M gene. Resistance to various antibiotics ranged from 18% to 77%. This study highlights the risk of virulent and multidrug-resistant STEC non-O157:H7 in raw milk and the need for proper quality surveillance and assurance plans to mitigate the potential public health threat.

The Influence of Lactobacillus spp. Secondary Metabolites Isolated from Immature Egyptian Honey on Human Pathogens, Transcription of Virulence Genes and Lung Cancer

This work aimed to isolate, and identify Lactic Acid Bacteria LAB from Egyptian immature citrus honey, and characterize their secondary metabolites, as well as determine the antibacterial activities and transcription of virulence genes (stx1, stx2, and eae) influenced by these bacterial secondary metabolites. From twenty hives, twenty immature citrus bee honey samples were taken. Traditional cultural and biochemical testing were used, followed by molecular confirmation. Further, LAB isolates' antibacterial and cytotoxic properties were investigated. 16S rRNA gene sequencing were assessed and, two lactic acid bacterial isolates were identified as Lactobacillus acidophilus Ch2 and Levilactobacillus brevis Ch1. Both isolates have good antagonistic action against clinical pathogens, with Levilactobacillus brevis Ch1 exhibiting the best antibacterial activity against all indicator pathogens examined. When compared to untreated cancer cells, the isolates demonstrated significant cytotoxic activity. Ch1 and Ch2 cell viability percentages were 39.5% and 18.76%, respectively. Furthermore, when exposed to Levilactobacillus brevis Ch1 metabolites, Shiga-producing Escherichia coli (STEC) virulence gene expression was suppressed. To identify bacterial secondary metabolites, a high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOF) approach was developed. Twenty-seven metabolites from diverse chemical classes were discovered in the crude extracts with antibacterial and anticancer characteristics. This is the first thorough investigation on the metabolic profile of LAB isolated from immature Egyptian honey and the findings suggested that isolates or their secondary metabolites could be used in the food sector as medicinal alternatives or as a biocontrol agent.

Prevalence of Salmonella spp. and Escherichia coli in the feces of free-roaming wildlife throughout South Korea.

Wildlife can carry pathogenic organisms, including viruses, bacteria, parasites, and fungi, which can spread to humans and cause mild to serious illnesses and even death. Spreading through animal feces, these pathogens significantly contributes to the global burden of human diseases. Therefore, the present study investigated the prevalence of zoonotic bacterial pathogens, such as Salmonella spp., Escherichia coli, and Shiga toxin-producing E. coli (STEC), in animal feces. Between September 2015 and August 2017, 699 wildlife fecal samples were collected from various agricultural production regions and mountainous areas in South Korea. Fecal samples were collected from wild mammals (85.26%, 596/699) and birds (14.73%, 103/699). Salmonella spp. and E. coli were present in 3% (21/699) and 45.63% (319/699) of the samples, respectively. Moreover, virulence genes stx1 and both stx1 and stx2 were detected in 13.30% (93/699) and 0.72% (5/699) of the samples, respectively. The 21 Salmonella spp. were detected in badgers (n = 5), leopard cats (n = 7), wild boars (n = 2), and magpies (n = 7); STEC was detected in roe deer, water deer, mice, and wild boars. Through phylogenetic and gene-network analyses, the Salmonella spp. isolates (n = 21 laboratory isolates, at least one isolate from each Salmonella-positive animal fecal sample, and n = 6 widely prevalent reference Salmonella serovars) were grouped into two major lineages: S. enterica subsp. enterica and S. enterica subsp. diarizonae. Similarly, 93 E. coli isolates belonged to stx1, including three major lineages (groups 1-3), and stx1 and stx2 detected groups. To the best of our knowledge, this is the first report of a wild leopard cat serving as a reservoir for Salmonella spp. in South Korea. The research findings can help manage the potential risk of wildlife contamination and improve precautionary measures to protect public health.

Detection and Molecular Characterization of Shiga-toxin-producing Escherichia coli from Fecal Samples of Cattle Farms Al-Buḥayrah Governorate Egypt

Escherichia coli that produce the Shiga toxin (STEC) has become one of the most significant foodborne pathogens linked to serious illnesses. Our study was aimed to detect the existence, antimicrobial profiles, and virulence repertoire of different STEC serotypes from fecal samples of dairy cattle.&#x0D; We collected 19 fecal samples (12 diarrheic and 7 non-diarrheic) from dairy cattle. E. coli isolation and identification were performed. Shiga toxins genes were detected and the antimicrobial resistance was investigated.&#x0D; E. coli was detected at the rate of 13/19 (68.4%) from the fecal samples. The frequency of Stx2a represented 12/13 (92.3%), Stx2 represented 3/13 (23.14%), and Stx1d, Stx2f, Stx2g, and eae represented 2/13 (15.4%). While the virulence genes Stx1, Stx1c, Stx2c, Stx2d, Stx2e, and ehxA represented 0/13 (0%). The most effective antimicrobials were tulathithromycin 30 (53.84%) followed by ceftiofur 30 and marbofloxacin (46.16%), amikacin (15.38%), and the least resistance was recorded for nalidixic acid and oxytetracycline (7.70%). The resistant antibiotics with percentages reached to 100% were Amoxicillin/ Clavulanic acid, Ceftriaxone, Ciprofloxacin, Doxycycline, Erythromycin, Gentamicin, Ninezolid, Nitrofurantoin, Penicillin G, Streptomycin, and Trimethoprim/Sulfamethoxazole, followed by chloramphenicol, nalidixic acid, oxytetracycline, and Tylosine (92.30%), Amikacin and Ampicillin (84.62%), Ceftiofur (23.07%), Marbofloxacin (15.38%) and Tulathithromycin 30 (7.70%).&#x0D; The findings demonstrated that E. coli that are resistant to numerous groups of antimicrobials spread through feces and may pose a risk to human and animal health.

The Pervasiveness of Bio-Hazards in Street-Vended Dairy Products

A total of 60 diverse dairy product samples, including ice cream, Ras cheese, and Kariesh cheese (20 of each), were randomly collected from street vendors in different regions of Alexandria governorate, Egypt to investigate the prevalence of pathogenic E. coli. The outcomes revealed presumptive positive colonies of 83.33% and 80% in ice cream and Kariesh cheese samples, respectively. Confirmatory tests via conventional biochemical identification affirmed the presence of E. coli in 80% of the samples. However, Vitek 2 compact testing revealed that only 46.66% were positive, in contrast to PCR techniques, which substantiated a prevalence of E. coli in only 25% of the samples, with 35% in street-vended Ras cheese and 25% in Kariesh cheese. The phoA gene, integral to E. coli, was prominently featured in street-vended Ras cheese samples at a striking 100%, while the virulence genes stx1 and stx2 were conspicuously absent, except for sporadic instances of the eaeA gene in street-vended products. These findings uncovered a nuanced landscape of bacterial presence, with E. coli exhibiting a significant prevalence disparity among product sources, particularly with street-vended items showing markedly high contamination rates. It is advised that awareness programs should be established to reduce dairy product contamination among street vendors. Keywords: Dairy; Street-vended; Escherichia coli; Contamination.

Molecular characterization of common zoonotic protozoan parasites and bacteria causing diarrhea in dairy calves in Ningxia Hui Autonomous Region, China.

Diarrhea caused by zoonotic pathogens is one of the most common diseases in dairy calves, threatening the health of young animals. Humans are also at risk, in particular children. To explore the pathogens causing diarrhea in dairy calves, the present study applied PCR-based sequencing tools to investigate the occurrence and molecular characteristics of three parasites (Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi) and three bacterial pathogens (Escherichia coli, Clostridium perfringens, and Salmonella spp.) in 343 fecal samples of diarrheic dairy calves from five farms in Lingwu County, Ningxia Hui Autonomous Region, China. The total positive rate of these pathogens in diarrheic dairy calves was 91.0% (312/343; 95% CI, 87.9-94.0), with C. perfringens (61.5%, 211/343; 95% CI, 56.3-66.7) being the dominant one. Co-infection with two to five pathogens was found in 67.3% (231/343; 95% CI, 62.4-72.3) of investigated samples. There were significant differences (p<0.05) in the positive rates of Cryptosporidium spp. and diarrheagenic E. coli among farms, age groups, and seasons. Two Cryptosporidium species (C. parvum and C. bovis) and five gp60 subtypes of C. parvum (IIdA15G1, IIdA20G1, IIdA19G1, IIdA14G1, and a novel IIdA13G1) were identified. Two assemblages (assemblage E and zoonotic assemblage A) of G. duodenalis and six ITS genotypes of E. bieneusi (J, Henan-IV, EbpC, I, EbpA, and ESH-01) were observed. Four virulence genes (eaeA, stx1, stx2, and st) of diarrheagenic E. coli and one toxin type (type A) of C. perfringens were detected. Our study enriches our knowledge on the characteristics and zoonotic potential of diarrhea-related pathogens in dairy calves.

Resistance and Virulence Surveillance in Escherichia coli Isolated from Commercial Meat Samples: A One Health Approach.

Escherichia coli is a key indicator of food hygiene, and its monitoring in meat samples points to the potential presence of antimicrobial-resistant strains capable of causing infections in humans, encompassing resistance profiles categorized as serious threats by the Centers for Disease Control and Prevention (CDC), such as Extended-Spectrum Beta-Lactamase (ESBL)-a problem with consequences for animal, human, and environmental health. The objective of the present work was to isolate and characterize ESBL-producing E. coli strains from poultry, pork, and beef meat samples, with a characterization of their virulence and antimicrobial resistance profiles. A total of 450 meat samples (150 chicken, 150 beef, and 150 pork) were obtained from supermarkets and subsequently cultured in medium supplemented with cefotaxime. The isolated colonies were characterized biochemically, followed by antibiogram testing using the disk diffusion technique. Further classification involved biofilm formation and the presence of antimicrobial resistance genes (blaCTX-M, AmpC-type, mcr-1, and fosA3), and virulence genes (eaeA, st, bfpA, lt, stx1, stx2, aggR, iss, ompT, hlyF, iutA, iroN, fyuA, cvaC, and hylA). Statistical analysis was performed via the likelihood-ratio test. In total, 168 strains were obtained, with 73% originating from chicken, 22% from pork, and 17% from beef samples. Notably, strains exhibited greater resistance to tetracycline (51%), ciprofloxacin (46%), and fosfomycin (38%), apart from β-lactams. The detection of antimicrobial resistance in food-isolated strains is noteworthy, underscoring the significance of antimicrobial resistance as a global concern. More than 90% of the strains were biofilm producers, and strains carrying many ExPEC genes were more likely to be biofilm formers (OR 2.42), which increases the problem since the microorganisms have a greater chance of environment persistence and genetic exchange. Regarding molecular characterization, bovine samples showed a higher prevalence of blaCTX-M-1 (OR 6.52), while chicken strains were more likely to carry the fosA3 gene (OR 2.43, CI 1.17-5.05) and presented between 6 to 8 ExPEC genes (OR 2.5, CI 1.33-5.01) compared to other meat samples. Concerning diarrheagenic E. coli genes, two strains harbored eae. It is important to highlight these strains, as they exhibited both biofilm-forming capacities and multidrug resistance (MDR), potentially enabling colonization in diverse environments and causing infections. In conclusion, this study underscores the presence of β-lactamase-producing E. coli strains, mainly in poultry samples, compared to beef and pork samples. Furthermore, all meat sample strains exhibited many virulence-associated extraintestinal genes, with some strains harboring diarrheagenic E. coli (DEC) genes.

Prevalence and Molecular Characterization of Shiga Toxin-Producing Escherichia coli from Food and Clinical Samples.

Shiga toxin-producing Escherichia coli (STEC) is one of the most prominent food-borne pathogens in humans. The current study aims to detect and to analyze the virulence factors, antibiotic resistance, and plasmid profiles for forty-six STEC strains, isolated from clinical and food strains. Pulsed-field gel electrophoresis (PFGE) was used to determine the genetic relatedness between different serotypes and sources of samples. The clinical samples were found to be resistant to Nb (100%), Tet (100%), Amp (20%), SXT (15%), and Kan (15%) antibiotics. In contrast, the food strains were found to be resistant to Nb (100%), Tet (33%), Amp (16.6%), and SXT (16.6%) antibiotics. The PFGE typing of the forty-six isolates was grouped into more than ten clusters, each with a similarity between 30% and 70%. Most of the isolates were found positive for more than five virulence genes (eae, hlyA, stx1, stx2, stx2f, stx2c, stx2e, stx2, nelB, pagC, sen, toxB, irp, efa, and efa1). All the isolates carried different sizes of the plasmids. The isolates were analyzed for plasmid replicon type by PCR, and 72.5% of the clinical isolates were found to contain X replicon-type plasmid, 50% of the clinical isolates contained FIB replicon-type plasmid, and 17.5% of the clinical isolates contained Y replicon-type plasmid. Three clinical isolates contained both I1 and Hi1 replicon-type plasmid. Only two food isolates contained B/O and W replicon-type plasmid. These results indicate that STEC strains have diverse clonal populations among food and clinical strains that are resistant to several antimicrobials. In conclusion, our findings indicate that food isolates of STEC strains harbor virulence, antimicrobial resistance, plasmid replicon typing determinants like those of other STEC strains from clinical strains. These results suggest that these strains are unique and may contribute to the virulence of the isolates. Therefore, surveillance and characterization of STEC strains can provide useful information about the prevalence of STEC in food and clinical sources. Furthermore, it will help to identify STEC serotypes that are highly pathogenic to humans and may emerge as a threat to public health.

Prevalence, characterization and antimicrobial resistance of non-O157 Shiga toxin-producing Escherichia coli isolates from healthy cattle in Tunisia

AbstractAs a continuation of our previous work, here we examined the occurrence, virulence factors and antibiotic resistance of non-O157 Shiga toxin-producing Escherichia coli (STEC) bacteria in faecal samples of healthy cattle in Tunisia. From 260 faecal samples, 236 E. coli isolates were obtained, of which 30 (12.7%) proved to be non-O157:H7 STEC. The presence of STEC-serogroup virulence genes stx1, stx2, eaeA and ehxA was detected by PCR. Further virulence genes and other genes used for phylogenetic grouping were also examined among the STEC isolates. The most dominant serogroup was O145, followed by O103 and O45. Shiga toxin gene (stx1) was found in 70% of the STEC strains, stx2 and ehxA were detected in 17 isolates (56.7%). Other virulence genes, including fimH, traT, sfa/focDE, iutA, cdt3, hly, fyuA and ibeA, were detected in 100%, 60%, 46.7%, 46.7%, 36.7%, 10%, 6.7% and 3.3%, of the strains, respectively. More than 70% of the isolates were susceptible to most of the tested antibiotics. We concluded that the presence of STEC O145, O103 and O45 (12.7%), together with the earlier described STEC O157:H7 (4.2%) in cattle, intended for slaughter, may represent a threat to public health. Efficient control measures are needed in the farms and slaughterhouses in Tunisia.

Shiga toxin-producing Escherichia coli (STEC) in meat and leafy greens available in the Swedish retail market – Occurrence and diversity of stx subtypes and serotypes

Shiga toxin-producing Escherichia coli (STEC) is a major cause of foodborne illness, ranging from mild diarrhea to permanent kidney failure. This study summarizes the results of four surveys performed at different time periods, which investigated the occurrence and characteristics of STEC in beef, lamb and leafy greens available in the Swedish retail market. Such data is required when assessing the public health risk of varying types of STEC in different foods, and for establishing risk management measures. Samples from domestic and imported products were collected based on their availability in the retail market. The occurrence of STEC was investigated in 477 samples of beef, 330 samples of lamb and 630 samples of leafy greens. The detection of virulence genes (stx1, stx2, eae) was performed using real-time PCR followed by the isolation of bacteria from stx-positive enriched samples using immunomagnetic separation or an immunoblotting method. All STEC isolated from the food samples was further characterised in terms of stx subtyping and serotyping through whole genome sequencing. STEC was isolated from 2 to 14 % of beef samples and 20 to 61 % of lamb samples, depending on the region of origin. STEC was not isolated from samples of leafy greens, although stx genes were detected in 11 (2 %) of the samples tested. In total, 5 of the 151 sequenced STEC isolates from meat contained stx2 and eae, of which 4 such combinations had the stx2a subtype. The stx2 gene, stx2a in particular, is strongly associated with serious disease in humans, especially in combination with the eae gene. The isolates belonged to 20 different serotypes. Two isolates from beef and one from lamb belonged to the serotype O157:H7 and contained genes for stx2 and eae. Overall, several combinations of stx subtypes were found in isolates from beef, whereas stx1c, either alone or together with stx2b, was the dominant combination found in STEC from lamb. In conclusion, STEC was rare in whole meat samples of domestic beef in the Swedish retail market, whereas such bacteria were frequently found in minced meat and whole meat samples of imported beef and domestic and imported lamb. Although the number of isolates containing genes linked to an increased risk of severe disease was low, beef and lamb in the Swedish retail market is a common source of human exposure to potentially pathogenic STEC.

Comparative Study on the Prevalence of Escherichia coli and it’s Antibiogram in Saum from Three Different Storage Places in Household at Mizoram

Background: Saum is a very important traditional food item of the Mizo society in Mizoram, North East India; but there is scanty scientific study and reports on Saum. The present study has been conducted to study and compare the prevalence of E. coli in Saum from three different storage places such as storage/keeping Saum under the sun (24-38°C) during hot part of the day, inside the refrigerator (3-4°C) and near the cooking fire (30-50°C) at home. Methods: The study was conducted from May, 2022 to November, 2022. A total of 105 Saum samples were collected from three different storage places comprising of 35 samples from each storage places and then analysed for the presence of foodborne pathogen E. coli and it’s virulence genes (stx1, stx2, elt, est genes) by conventional method and PCR. All the E. coli isolates were subjected to 12 different antibiotics for antimicrobial resistance pattern by disc diffusion method. Result: The overall prevalence of E. coli from 105 samples was 7.62% (8/105) comprising of 5.71% (2/35) from the storage place under the sun, 2.86% (1/35) from the refrigerator and 14.29% (5/35) near the cooking fire and no presence of virulence genes of E. coli was detected in the present study. The antimicrobial resistance pattern of all the 8 E. coli isolates showed the highest resistance to Cefazolin (100%) followed by Imipenem (37.5%). The present work will be a complementary contribution among Mizo society to know the best and safest storage places of Saum in regards to public health point of view.

First molecular detection of shiga toxin-producing Escherichia coli in dogs from serbia: a potential threat to human health?

Shiga toxin‑producing Escherichia coli (STEC) are considered one of the most significant E. coli pathotypes transmitted by food, causing life‑threatening conditions in children and elderly people. The aim of this study was to investigate the presence and determine the prevalence of STEC in dogs in Serbia by conventional PCR method, targeting three major virulence genes (stx1, stx2, and eae). The overall percentage of positive samples was 12.87% (13/101), with the stx2 gene, the more potent of the two toxins, found in all the positive samples. The finding of eae gene in combination with stx genes (8/13) within the same genetic pool implies the potential presence of enterohemorrhagic E. coli or the potential emergence of these strains, considering an efficient mechanism of horizontal transfer of three major virulence genes. Our results also highlight dogs' lifestyle as a risk factor for STEC colonisation. These E. coli strains, according to our results, are more likely to be found in dogs living outdoors than those kept in house. Due to significant prevalence of STEC in dogs determined in this research and due to close contact between dogs and humans, dogs could be considered a source of human infections.

Prevalence and In Vivo Assessment of Virulence in Shiga Toxin-Producing Escherichia coli Clinical Isolates from Greater Cairo Area.

Background: Shiga toxin-producing Escherichia coli (STEC) has been identified as an important etiologic agent of human disease in Egypt. Aims: To investigate the occurrence and describe the characterization as well as prevalence of STEC in Greater Cairo hospitals as well as molecular characterization of virulence and resistance genes. Methods: Four hundred seventy E. coli clinical isolates were collected from eight hospitals and analyzed by genotypic and phenotypic methods for STEC, followed by histopathological examination and scoring of different organs lesions. Results: The highest proportion of isolates was from urine (151 isolates), whereas the lowest was from splenic drain (3 isolates). In tandem, when serogrouping was performed, 15 serogroups were obtained where the most prevalent was O157 and the least prevalent was O151. All isolates were positive when screened for identity gene gad A, while only typable strains were screened for seven virulence genes stx1 (gene encoding Shiga toxin 1), stx2 (gene encoding Shiga toxin 2), tsh (gene encoding thermostable hemagglutinin), eaeA (gene encoding intimin), invE (gene encoding invasion protein), aggR (gene encoding aggregative adherence transcriptional regulator), and astA (aspartate transaminase) where the prevalence was 48%, 30%, 50%, 57%, 7.5%, 12%, and 58%, respectively. Of 254 typable isolates, 152 were STEC carrying stx1 or stx2 genes or both. Conclusions: Relying on in vivo comparison between different E. coli pathotypes via histopathological examination of different organs, E. coli pathotypes could be divided into mild virulent, moderate virulent, and high virulent strains. Statistical analysis revealed significant correlation between different serogroups and presence of virulence genes.

Detection of Virulence Genes Stx1, Stx2, eae A and hly A in E.coli Isolated from Local Minced Meat

The current study included the investigation of E. coli bacteria in random samples of ground meat, the detection of virulence genes in isolates, and the study of isolates resistance to some widely used antibiotics. Twenty-four samples of minced meat were collected to isolate and identify E. coli bacteria by phenotypic, microscopic characteristics and traditional biochemical tests were done. Antibiotic susceptibility tests, genomes of isolates were extracted, the virulence factors Stx1, Stx2, eae A and hly A were investigated by multiplex PCR technique. 7 isolates showed Colonies with a metallic green luster on EMB medium, gram negative, rod shaped bacteria, facultative anaerobic bacteria, positive for indole, methyl red and catalase tests, and negative for Fox Proscore, urease and oxidase tests. All isolates were containing hly A gene, 6 isolates were containing the virulence gene Stx1, 5 isolates containing were eae A gene and 4 isolates were containing Stx2 gene. The results shown a high resistance of E. coli isolates to all types of antibiotics used, as all isolates were possessed multiple antibiotic resistance.

Rapid detection of Shiga-toxin-producing Escherichia coli O157:H7 based on a colorimetric loop-mediated isothermal amplification (cLAMP) assay using a molecular beacon paired with HRPzyme.

Contamination by Escherichia coli O157:H7 is considered a threat in the livestock and food industries. Therefore, it is necessary to develop methods for the convenient and rapid detection of Shiga-toxin-producing E. coli O157:H7. This study aimed to develop a colorimetric loop-mediated isothermal amplification (cLAMP) assay using a molecular beacon to rapidly detect E. coli O157:H7. Primers and a molecular beacon were designed for targeting the Shiga-toxin-producing virulence genes (stx1 and stx2) as molecular markers. Additionally, Bst polymerase concentration and amplification conditions for bacterial detection were optimized. The sensitivity and specificity of the assay were also investigated and validated on artificially tainted (100-104 CFU/g) Korean beef samples. The cLAMP assay could detect 1 × 101 CFU/g at 65 °C for both genes, and the assay was confirmed to be specific for E. coli O157:H7. The cLAMP takes about an hour and does not require expensive devices (e.g., thermal cycler and detector). Hence, the cLAMP assay proposed herein can be used in the meat industry as a fast and simple way to detect E. coli O157:H7.

Phenotypic and genotypic aspects of sorbitol-negative Escherichia coli isolated from cattle

Escherichia coli especially sorbitol-negative serogroups such as O157 that produces Shiga toxin (STEC) is involved in food poisoning with severity ranging from individual cases into outbreaks that threaten human and animal health. This study aimed to evaluate phenotypic and genotypic aspects of sorbitol-negative E.coli isolated from different sources. A total of 420 samples were collected from fecal swabs and raw milk of cattle from various locations in Kafr Elsheikh government, Egypt. The prevalence of E. coli isolates was 9.3% (39/420) as was determined by bacterial culturing. The phenotype of these isolates was serologically and molecularly determined, and the results showed an overall prevalence of E. coli O157 of 23% (9/39) with a high incidence in fecal swabs samples. The multidrug resistance (MDR) phenotype as detected by antibiotic sensitivity test and confirmed by PCR revealed resistance to amoxicillin, clavulanic acid, vancomycin, ampicillin, and sulbactam with the detection of blaTEM and blaSHV MDR genes in 9 and 2 isolates, respectively. Virulence genes (stx1, stx2, eaeA) were also detected in 1, 9, 6 isolates, respectively. With these results, we could conclude that E. coli O157 was identified in not only cattle fecal swab samples but also in their milk and subsequently this could threaten animal and human health.

SQUAB AND QUAIL MEATS: MICROBIAL STATUS AND PREVALENCE OF MULTIDRUG-RESISTANT SHIGA TOXIN-PRODUCING E. coli

The consumption of exotic meats, such as squab and quail meats, is common in many parts of the world. However, little is known about the safety of these foods. This study aimed to evaluate the microbial characteristics of squab and quail meats, with a particular interest in the prevalence of Shiga toxin-producing Escherichia coli (STEC). Meat samples were examined for total bacterial counts, total mold counts, most probable number of coliforms, total E. coli counts, and the prevalence of STEC. The presence of virulence genes (stx1, stx2, and eae) in STEC isolates was also investigated. Results from microbial analyses revealed poor hygienic status of squab and quail meats. E. coli was isolated from 16% and 10% of squab and quail meats, respectively. Six E. coli serotypes were recovered, including O26, O78, O111, O114, O119, and O127, and STEC genes were detected in all these isolates. Squab liver had the highest E. coli prevalence rates, followed by gizzard, heart, spleen, and breast muscles. The prevalence of E. coli in quail meat samples was similar across all tissues. STEC serotypes showed notable multidrug resistance profiles. We then used ascorbic and rosmarinic acids to increase the safety of breast muscle. Treatment of breast muscles with these acids significantly improved their microbial safety. These findings highlight the potential role of squab and quail meats as a vehicle for STEC transmission to humans, and the beneficial effect of treatment with ascorbic and rosmarinic acids on enhancing the safety of exotic meats. Key words: food safety; shiga toxin; E. coli; squab; quails; meat