Biofilm-producing Escherichia coli O104:H4 overcomes bile salts toxicity by expressing virulence and resistance proteins.

We investigated bile salts' ability to induce phenotypic changes in biofilm production and protein expression of pathogenic Escherichia coli strains. For this purpose, 82 pathogenic E. coli strains isolated from humans (n = 70), and animals (n = 12), were examined for their ability to form biofilms in the presence or absence of bile salts. We also identified bacterial proteins expressed in response to bile salts using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-electrophoresis) and liquid chromatography-mass spectrometry (LC-MS/MS). Lastly, we evaluated the ability of these strains to adhere to Caco-2 epithelial cells in the presence of bile salts. Regarding biofilm formation, two strains isolated from an outbreak in Republic of Georgia in 2009 were the only ones that showed a high and moderate capacity to form biofilm in the presence of bile salts. Further, we observed that those isolates, when in the presence of bile salts, expressed different proteins identified as outer membrane proteins (i.e. OmpC), and resistance to adverse growth conditions (i.e. F0F1, HN-S, and L7/L12). We also found that these isolates exhibited high adhesion to epithelial cells in the presence of bile salts. Together, these results contribute to the phenotypic characterization of E. coli O104: H4 strains.

Optimizing the Antimicrobial Activity of Sodium Hypochlorite (NaClO) over Exposure Time for the Control of Salmonella spp. In Vitro.

Fish is a nutritionally rich product; however, it is easily contaminated by pathogenic microorganisms, such as Salmonella spp. Therefore, this study aimed to identify the best concentration of sodium hypochlorite (NaClO), exposure time, and water temperature that allow the most effective antimicrobial effect on the viable population of Salmonella spp. Thus, Salmonella Enteritidis ATCC 13076 and Salmonella Schwarzengrund were exposed to different time frames, ranging from 5 min to 38.5 min, temperatures between 5 and 38.5 °C, and NaClO concentrations ranging from 0.36 to 6.36 ppm, through a central composite rotational design experiment (CCRD). The results demonstrated that the ATCC strain exhibited a quadratic response to sodium hypochlorite when combined with exposure time, indicating that initial contact would already be sufficient for the compound's action to inhibit the growth of the mentioned bacteria. However, for S. Schwarzengrund (isolated directly from fish cultivated in aquaculture), both NaClO concentration and exposure time significantly influenced inactivation, following a linear pattern. This suggests that increasing the exposure time of NaClO could be an alternative to enhance Salmonella elimination rates in fish slaughterhouses. Thus, the analysis indicates that the Salmonella spp. strains used in in vitro experiments were sensitive to concentrations equal to or greater than the recommended ones, requiring a longer exposure time combined with the recommended NaClO concentration in the case of isolates from aquaculture.

Can Extraintestinal Pathogenic Escherichia coli with Heat Resistance Profile Overcome Nonthermal Technologies?

Ultraviolet-C light-emitting diode (UVC-LED) and ultrasound (US) are two nonthermal technologies with the potential to destroy pathogens. However, little is known about their effectiveness in strains with a history of heat resistance. Thus, this study aimed to evaluate the phenotype and genotype of heat-resistant extraintestinal pathogenic Escherichia coli (ExPEC) with heat resistance genes after the application of US, UVC-LED, and UVC-LED+US. For this, two central composite rotatable designs were used to optimize the UVC-LED and US conditions in four ExPEC isolated from beef. From the genome of these isolates obtained in a previous study, possible genes for UVC resistance were analyzed. Results showed that US was ineffective in reducing >0.30 log colony-forming unit/mL, and that when used after UVC-LED, it showed a nonsynergic or antagonistic effect. Also, UVC-LED had the greatest effect at the maximum dose (4950 mJ/cm2 from 1.65 mW/cm2 for 50 min). However, the strains showed some recovery after that, which could be implicated in the expression of genes included in SOS system genes, some others present in the transmissible Locus of Stress Tolerance (trxBC and degP), and others (terC). Thus, ExPEC can overcome the conditions used in this study for US, UVC-LED, and UVC-LED+US, probably due to the history of resistance to other cellular damage. The result of this study will contribute to future studies that aim to find better treatment conditions for each food product.

Genomic features and heat resistance profiles of Escherichia coli isolated from Brazilian beef.

AIMS: Characterize Escherichia coli and E. coli -producing (STEC) isolates from Brazilian beef to determine heat resistance and the presence of the transmissible locus of stress tolerance (tLST). METHODS AND RESULTS: Twenty-two STEC previously isolated from beef and characterized as STEC by PCR were subjected to different heat survival challenges (60°C and 71°C). Furthermore, the three tLST-positive isolates and one tLST-negative isolate by PCR were selected for WGS analysis. Phenotypic results indicated that 3/22 (13.64%) were heat resistant, 12/22 (54.54%) were moderately resistant, and 7/22 (31.82%) were sensitive to heat treatments. WGS analyses showed that three isolates with heat resistance showed tLST with up to 80% and 42% of similarity by BLAST analysis, with the major tLST genes being responsible for the homeostasis module. However, WGS showed the absence of stx genes associated with tLST-positive isolates, albeit with virulence and resistance genes found in extraintestinal pathogenic E. coli (ExPEC). CONCLUSION: Our findings demonstrate the presence of heat-resistant E. coli as well as confirm some tLST genes in E. coli isolated from Brazilian beef.

Heat-resistant and biofilm-forming Escherichia coli in pasteurized milk from Brazil.

Escherichia coli harboring a transmissible locus of stress tolerance (tLST) and the ability to form biofilms represent a serious risk in dairy production. Thus, we aimed to evaluate the microbiological quality of pasteurized milk from two dairy producers in Mato Grosso, Brazil, with a focus on determining the possible presence of E. coli with heat resistance (60 °C/6 min), biofilm-forming potential phenotypes and genotypes, and antimicrobial susceptibility. For this, fifty pasteurized milk samples from producers named A and B were obtained for 5 weeks to investigate the presence of Enterobacteriaceae members, coliforms, and E. coli. For heat resistance, E. coli isolates were exposed to a water bath at 60 °C for 0 and 6 min. In antibiogram analysis, eight antibiotics belonging to six antimicrobial classes were analyzed. The potential to form biofilms was quantified at 570 nm, and curli expression by Congo Red was analyzed. To determine the genotypic profile, we performed PCR for the tLST and rpoS genes, and pulsed-field gel electrophoresis (PFGE) was used to investigate the clonal profile of the isolates. Thus, producer A presented unsatisfactory microbiological conditions regarding Enterobacteriaceae and coliforms for weeks 4 and 5, while all samples analyzed for producer B were contaminated at above-the-limit levels established by national and international legislation. These unsatisfactory conditions enabled us to isolate 31 E. coli from both producers (7 isolates from producer A and 24 isolates from producer B). In this way, 6 E. coli isolates (5 from producer A and 1 from producer B) were highly heat resistant. However, although only 6 E. coli showed a highly heat-resistant profile, 97% (30/31) of all E. coli were tLST-positive. In contrast, all isolates were sensitive to all antimicrobials tested. In addition, moderate or weak biofilm potential was verified in 51.6% (16/31), and the expression of curli and presence of rpoS was not always related to this biofilm potential. Therefore, the results emphasize the spreading of heat-resistant E. coli with tLST in both producers and indicate the biofilm as a possible source of contamination during milk pasteurization. However, the possibility of E. coli producing biofilm and surviving pasteurization temperatures cannot be ruled out, and this should be investigated.

Genomic analysis of Mycobacterium tuberculosis variant bovis strains isolated from bovine in the state of Mato Grosso, Brazil.

The species Mycobacterium tuberculosis variant bovis (M. tuberculosis var. bovis) is associated with tuberculosis, mainly in cattle and buffaloes. This pathogen has the potential to infect other mammals, including humans. Tuberculosis caused by M. tuberculosis var. bovis is a zoonosis clinically identical to tuberculosis caused by Mycobacterium tuberculosis, and the recommended treatment in humans results in the use of antibiotics. In this study, we used the whole genome sequencing (WGS) methodology Illumina NovaSeq 6000 System platform to characterize the genome of M. tuberculosis var. bovis in cattle circulating in Mato Grosso, identify mutations related to drug resistance genes, compare with other strains of M. tuberculosis var. bovis brazilian and assess potential drug resistance. Four isolates of M. tuberculosis var. bovis of cattle origin representing the main livestock circuits, which had been more prevalent in previous studies in the state of Mato Grosso, were selected for the genomic study. The genome sizes of the sequenced strains ranged from 4,306,423 to 4,332,964 bp, and the GC content was 65.6%. The four strains from Mato Grosso presented resistance genes to pncA (pyrazinamide), characterized as drug-resistant strains. In addition to verifying several point mutations in the pncA, rpsA, rpsL, gid, rpoB, katG, gyrB, gyrA, tlyA, embA, embB, embC, fgd, fbiB, and fbiC genes, these genes were similar to antibiotic resistance in more than 92% of the Brazilian strains. Therefore, our results indicated a high genetic diversity between our isolates and other M. tuberculosis var. bovis isolated in Brazil. Thus, multiple transmission routes of this pathogen may be present in the production chain. So, to achieve a bovine tuberculosis-free health status, the use of the WGS as a control and monitoring tool will be crucial to determine these transmission routes.

Salmonella Behavior in Meat during Cool Storage: A Systematic Review and Meta-Analysis.

The aim of the present study was to investigate Salmonella behavior in meat stored in cool conditions (between 0 °C and 7.5 °C), by employing a systematic review and meta-analysis. The data were obtained from research articles published in SciELO, PubMed, the Web of Science, and Scopus databases. The results of the retrieved studies were obtained from meat (beef, chicken, pork, poultry, and turkey), fish, shellfish, and broth media samples The data were extracted as sample size (n), initial concentration (Xi), final concentration (Xf), standard deviation (SD), standard error (SE), and microbial behavior effects (reduction or growth). A meta-analysis was carried out using the metaphor package from R software. A total of 654 articles were initially retrieved. After applying the exclusion criteria, 83 articles were selected for the systematic review, and 61 of these were used for the meta-analysis. Most studies were conducted at 0 °C to 4.4 °C storage temperatures under normal atmosphere package conditions. Salmonella Typhimurium, S. Enteritidis, and a cocktail (strain mixture) were inoculated at 5.0 and 6.0 log CFU mL−1. Articles both with and without the addition of antimicrobial compounds were found. Salmonella concentration decreases were observed in most studies, estimated for all study combinations as −0.8429 ± 0.0931 log CFU g−1 (95% CI; −1.0254, −0.6604) (p < 0.001), varying for each subgroup analysis. According to this survey, Salmonella concentration decreases are frequent during cool storage, although concentration increases and no bacterial inactivation were observed in some studies.

Antimicrobial Resistance of Listeria monocytogenes from Animal Foods to First- and Second-Line Drugs in the Treatment of Listeriosis from 2008 to 2021: A Systematic Review and Meta-Analysis.

First-line drugs for the treatment of listeriosis are the same around the world, but particular conditions might reduce their efficacy, including antimicrobial resistance. Therefore, this study aimed to verify, based on a systematic review and meta-analysis, whether the prevalence of antimicrobial resistance in Listeria monocytogenes from animal foods is higher for first- or second-line antimicrobials. From the total of 302 identified studies, 16 met all the eligibility criteria from 2008 to 2021 and were included in this meta-analysis. They comprised a dataset of 1152 L. monocytogenes isolates, obtained from different animal food products, food processing environment, and live animals. The included studies were developed in South America (n = 5), Europe (n = 4), Asia (n = 3), Africa (n = 2), and North America (n = 2), testing a total of 35 different antimicrobials, 11 of them classified as first-line drugs. Complete lack of antimicrobial resistance across the studies (all L. monocytogenes isolates tested as susceptible) was only observed for linezolid, while widespread antimicrobial resistance (all L. monocytogenes isolates tested resistant) was described for amoxicillin, benzylpenicillin, cefoxitin, fusidic acid, imipenem, sulfamethoxazole, and vancomycin. Overall, the meta-analysis results indicated no evidence that antimicrobial resistance in L. monocytogenes isolated from animal-based food is higher for first-line antimicrobials compared to second-line compounds (p=0.37). A greater volume of publication, together with better characterization of the isolates, is still needed for a more precise estimate of the real prevalence of antimicrobial resistance in L. monocytogenes.

Whole-Genome Sequencing Analyses of Heat-Resistant Escherichia coli Isolated from Brazilian Beef.

Four Escherichia coli isolates with moderate or high heat resistance were sequenced. Through sequencing, truncated transmissible locus of stress tolerance (tLST) variants tLST1 and tLSTa were identified in the three isolates. The most identified tLST genes (clpK and hsp) are responsible for the homeostasis module.

Technical recommendations for updating good food manufacturing practices (GMP) aimed at preventing COVID-19 in food services: checklist and review of GMP manual / Recomendações técnicas para atualização das boas práticas de fabricação de alimentos (BPF) visando a prevenção da COVID-19 em serviços de alimentação: checklist e revisão de manuais BPF

The COVID-19 pandemic caused by the SARS-CoV-2 virus brought additional challenges to the Food and Nutrition Service, so in addition to meeting the hygienic-sanitary conditions of food, companies now need to incorporate new manufacturing practices. foods that aim to preserve the health of workers as well as consumers. In this context, this study aimed to carry out a literature review on technical recommendations for good food manufacturing practices related to the prevention of COVID-19 and prepare a checklist to facilitate the identification of failures, assess risk COVID-19 transmission in food services and guide how to adapt the good manufacturing practices manuals. Based on technical notes (NT) 47, 48 and 49 of 2020 that were edited by ANVISA, based on the standards of the World Health Organization (WHO), a questionnaire was created containing 100 questions in the various categories of the food production sector, such as physical structure/building, hand washing conditions, general worker protection measures, material storage area, personal hygiene conditions, food storage area, company personnel, customer service area, preparation area, portioning and distribution area, food consumption area for workers, area for reception and food services received and food delivery services. In this way, we understand that biosafety measures must be adopted from the update of the manuals of good manufacturing practices in food services, and, to support this action, we propose the use of the checklist in the appendix to identify non-conformities related to the prevention of COVID -19.

A pandemia da COVID-19 causada pelo vírus SARS-CoV-2 trouxe desafios adicionais ao Serviço de Alimentação e Nutrição (SAN), pois, além de atender as condições higiênicas sanitárias dos alimentos, as empresas agora precisam incorporar novas práticas que visem preservar a saúde das pessoas. Nesse contexto, esse estudo buscou realizar uma revisão da literatura sobre as recomendações técnicas de boas práticas de fabricação de alimentos relacionadas com a prevenção da COVID-19 e elaborar uma lista de verificações (check-list) para facilitar a identificação de falhas, avaliar risco de transmissão da COVID-19 nos serviços de alimentação e orientar as adaptações dos manuais de boas práticas de fabricação. Com base nas notas técnicas vigentes no Brasil e baseadas nas diretrizes da Organização Mundial de Saúde (OMS), foi elaborado um questionário contendo 100 perguntas nas diversas categorias do setor de produção de alimentos. Dessa maneira, entendemos que medidas de biossegurança devem ser adotadas a partir da atualização dos manuais de boas práticas de fabricação nos serviços de alimentação, e, para apoiar essa ação, propomos a utilização do checklist em apêndice para identificar as inconformidades relacionadas a prevenção da COVID-19.

Salmonella spp. in Aquaculture: An Exploratory Analysis (Integrative Review) of Microbiological Diagnoses between 2000 and 2020.

The present study aimed to characterize, through descriptive statistics, data from scientific articles selected in a systematic integrative review that performed a microbiological diagnosis of Salmonella spp. in aquaculture. Data were obtained from research articles published in the BVS, Scielo, Science Direct, Scopus and Web of Science databases. The selected studies were published between 2000 and 2020 on samples of aquaculture animal production (fish, shrimp, bivalve mollusks, and other crustaceans) and environmental samples of aquaculture activity (farming water, soil, and sediments). After applying the exclusion criteria, 80 articles were selected. Data such as country of origin, categories of fish investigated, methods of microbiological diagnosis of Salmonella spp., sample units analyzed and most reported serovars were mined. A textual analysis of the word cloud and by similarity and descending hierarchical classification with the application of Reinert's algorithm was performed using R® and Iramuteq® software. The results showed that a higher percentage of the selected articles came from Asian countries (38.75%). Fish was the most sampled category, and the units of analysis of the culture water, muscle and intestine were more positive. The culture isolation method is the most widespread, supported by more accurate techniques such as PCR. The most prevalent Salmonella serovars reported were S. Typhimurium, S. Weltevreden and S. Newport. The textual analysis showed a strong association of the terms "Salmonella", "fish" and "water", and the highest hierarchical class grouped 25.4% of the associated text segments, such as "aquaculture", "food" and "public health". The information produced characterizes the occurrence of Salmonella spp. in the aquaculture sector, providing an overview of recent years. Future research focusing on strategies for the control and prevention of Salmonella spp. in fish production are necessary and should be encouraged.

Inconsistent PCR detection of Shiga toxin-producing Escherichia coli: Insights from whole genome sequence analyses.

Shiga toxin-producing Escherichia coli (STEC) have been linked to food-borne disease outbreaks. As PCR is routinely used to screen foods for STEC, it is important that factors leading to inconsistent detection of STEC by PCR are understood. This study used whole genome sequencing (WGS) to investigate causes of inconsistent PCR detection of stx1, stx2, and serogroup-specific genes. Fifty strains isolated from Alberta feedlot cattle from three different studies were selected with inconsistent or consistent detection of stx and serogroup by PCR. All isolates were initially classified as STEC by PCR. Sequencing was performed using Illumina MiSeq® with sample library by Nextera XT. Virtual PCRs were performed using Geneious and bacteriophage content was determined using PHASTER. Sequencing coverage ranged from 47 to 102x, averaging 74x, with sequences deposited in the NCBI database. Eleven strains were confirmed by WGS as STEC having complete stxA and stxB subunits. However, truncated stx fragments occurred in twenty-two other isolates, some having multiple stx fragments in the genome. Isolates with complete stx by WGS had consistent stx1 and stx2 detection by PCR, although one also having a stx2 fragment had inconsistent stx2 PCR. For all STEC and 18/39 non-STEC, serogroups determined by PCR agreed with those determined by WGS. An additional three WGS serotypes were inconclusive and two isolates were Citrobacter spp. Results demonstrate that stx fragments associated with stx-carrying bacteriophages in the E. coli genome may contribute to inconsistent detection of stx1 and stx2 by PCR. Fourteen isolates had integrated stx bacteriophage but lacked complete or fragmentary stx possibly due to partial bacteriophage excision after sub-cultivation or other unclear mechanisms. The majority of STEC isolates (7/11) did not have identifiable bacteriophage DNA in the contig(s) where stx was located, likely increasing the stability of stx in the bacterial genome and its detection by PCR.

Inactivation of Multi-Drug Resistant Non-Typhoidal Salmonella and Wild-Type Escherichia coli STEC Using Organic Acids: A Potential Alternative to the Food Industry.

Salmonella and Escherichia coli are the main bacterial species involved in food outbreaks worldwide. Recent reports showed that chemical sanitizers commonly used to control these pathogens could induce antibiotic resistance. Therefore, this study aimed to describe the efficiency of chemical sanitizers and organic acids when inactivating wild and clinical strains of Salmonella and E. coli, targeting a 4-log reduction. To achieve this goal, three methods were applied. (i) Disk-diffusion challenge for organic acids. (ii) Determination of MIC for two acids (acetic and lactic), as well as two sanitizers (quaternary compound and sodium hypochlorite). (iii) The development of inactivation models from the previously defined concentrations. In disk-diffusion, the results indicated that wild strains have higher resistance potential when compared to clinical strains. Regarding the models, quaternary ammonium and lactic acid showed a linear pattern of inactivation, while sodium hypochlorite had a linear pattern with tail dispersion, and acetic acid has Weibull dispersion to E. coli. The concentration to 4-log reduction differed from Salmonella and E. coli in acetic acid and sodium hypochlorite. The use of organic acids is an alternative method for antimicrobial control. Our study indicates the levels of organic acids and sanitizers to be used in the inactivation of emerging foodborne pathogens.

Behavior of spoilage bacteria and Salmonella enterica subspecies enterica O:4,5 in vacuum-packaged beef during refrigeration / Comportamento de bactérias deteriorantes e Salmonella enterica subespécie entérica O:4,5 em carne bovina embalada a vácuo durante refrigeração

ABSTRACT: In this study, the kinetic parameters of mesophilic, psychrotrophic and lactic acid bacteria in vacuum-packed beef at 1 °C and 4 °C were estimated from experimental growth curves produced by samples stored during 21 and 60 days, respectively. In a separate experiment, the survival of multidrug resistant (MDR) Salmonella enterica O:4,5 at 1°C was also characterized. The shelf-life of vacuum-packed beef stored at 4 °C was estimated at 16.1 days (95% CI: 14.8 - 17.3 days), whereas at 1 °C it was longer than 21 days because the mesophiles count estimated towards the end of the experiment was 12.5 ln CFU.g-1 (95% CI: 11.8 - 13.3 ln CFU.g-1) which is lower than the shelf-life reference value. At 1 °C, inoculated Salmonella was reduced in 6.61 ln CFU.g-1 (2.87 log CFU.g-1). These results demonstrated the importance of establishing in legislation, especially in Brazil, standard values of deteriorating microorganisms in beef for maintaining product quality.

RESUMO: Neste estudo, os parâmetros cinéticos de bactérias mesófilas, psicrotróficas e ácido lácticas foram estimados em carne bovina embalada a vácuo a 1 °C e 4 °C, a partir de curvas experimentais produzidas em amostras estocadas durante 21 e 60 dias, respectivamente. Em um experimento separado, a sobrevivência de Salmonella enterica O:4,5 multirresistente (MDR) a 1°C também foi caracterizada. A vida de prateleira da carne bovina embalada a vácuo, estocada a 4°C, foi estimada em 16.1 dias (95% CI: 14.8 - 17.3 dias), enquanto que a 1 °C o período foi maior que 21 dias, porque a contagem estimada de mesófilos ao final do experimento foi de 12.5 ln UFC.g-1 (95% CI: 11.8 - 13.3 ln UFC.g-1), o qual é mais baixo que o valor referência de shelf-life. A 1 °C, Salmonella inoculada reduziu em 6.61 ln UFC.g-1 (2.87 log UFC.g-1). Estes resultados demonstram a importância de estabelecimento em legislação, especialmente no Brasil, de valores padrões para contagem de microrganismos deteriorantes em carnes visando manter a qualidade do produto.

Acetic Acid Increased the Inactivation of Multi-drug Resistant Non-typhoidal Salmonella by Large-Scaffold Antibiotic.

Salmonella is a gram-negative bacterium with intrinsic resistance to large-scaffold antibiotics due to the presence of an outer membrane. Based on the mode of action of the organic acids in outer membrane disintegration, and consequently, an enhancement in cell permeability, a combination of acetic acid and a large-scaffold antibiotic is it evaluated. Therefore, the aim of this study is to assess the combination of different levels of acetic acid with vancomycin, in order to determine whether or not the organic acid may overcome the cell wall and the intrinsic resistance in multi-drug resistant Salmonella. Screening of five wild-type Salmonella strains and one clinical strain was performed to select the strain more resistance to acid inhibition. Acetic acid was tested at 2.0, 1.75, 1.50, and 1.25% levels, separated or combined with 8 µg/mL vancomycin dose. An aliquot was collected after exposure and inoculated into the brain and heart infusion agar. The plates were counted and the data analyzed by ANOVA and a posthoc Tukey test (p < 0.05). The results indicate that 1.25 and 1.50% levels did not affect the vancomycin inactivation of multi-drug resistant Salmonella. However, at levels of 1.75 and 2.0%, an increase in microbial reduction is observed. Also, 2% level acetic acid and vancomycin had a threefold increase compared to vancomycin alone. Therefore, the use of acetic acid as prior treatment for Salmonella increased the inactivation rate of vancomycin. The combination of organic acid and antibiotics is a potential tool to overcome cases of antimicrobial resistance.

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Shiga-Toxin Producing Escherichia Coli in Brazil: A Systematic Review.

Shiga-toxin producing E. coli (STEC) can cause serious illnesses, including hemorrhagic colitis and hemolytic uremic syndrome. This is the first systematic review of STEC in Brazil, and will report the main serogroups detected in animals, food products and foodborne diseases. Data were obtained from online databases accessed in January 2019. Papers were selected from each database using the Mesh term entries. Although no human disease outbreaks in Brazil related to STEC has been reported, the presence of several serogroups such as O157 and O111 has been verified in animals, food, and humans. Moreover, other serogroups monitored by international federal agencies and involved in outbreak cases worldwide were detected, and other unusual strains were involved in some isolated individual cases of foodborne disease, such as serotype O118:H16 and serogroup O165. The epidemiological data presented herein indicates the presence of several pathogenic serogroups, including O157:H7, O26, O103, and O111, which have been linked to disease outbreaks worldwide. As available data are concentrated in the Sao Paulo state and almost completely lacking in outlying regions, epidemiological monitoring in Brazil for STEC needs to be expanded and food safety standards for this pathogen should be aligned to that of the food safety standards of international bodies.

Occurrence and antimicrobial resistance of E. coli non-O157 isolated from beef in Mato Grosso, Brazil.

Shiga toxin-producing Escherichia coli (STEC) is an important public health concern pathogen, as it produces two toxins, Stx1 and Stx2, with cytotoxic capacity. In addition, STEC strains are frequently involved in food outbreaks worldwide, leading to public health challenges and economic losses. In this context, the occurrence and antimicrobial resistance profile of the STEC isolated from fresh beef produced in Mato Grosso, Brazil, were estimated. One hundred seven retail beef under vacuum-packaged produced by 13 different slaughterhouses were submitted to microbiological, molecular, and antimicrobial resistance analyses. STEC occurrence in beef was of 4.67%, and five strains presented the stx2 gene. The O111 serogroup, reported in several outbreak cases worldwide, was detected, and other serotypes (O8:H20, O22:H16, and O141:H49) were also isolated. All isolated strains displayed sensitivity to 12 antibiotics, except for two strains, which where streptomycin-resistant. The presence of STEC in retail beef samples indicates public health risks with significant economic impact throughout the retail beef chain.

Escherichia coli O26 and O113:H21 on Carcasses and Beef from a Slaughterhouse Located in Mato Grosso, Brazil.

Shiga toxin-producing Escherichia coli (STEC) is a group of emerging pathogens that can cause human diseases, including hemolytic uremic syndrome (HUS) and hemorrhagic colitis (HC). Monitoring slaughtering stages and checking contamination points are crucial for the production of safe food. In this context, the aim of this study was to verify contamination by STEC strains, to determine the contamination points and evaluate the resistance profile to 12 antimicrobials used in both veterinary and human medicine. A total of 80 samples were obtained from eight collection points (pen floor, rectum, hide, carcass swabs and esophagus, diaphragm, masseter, and retail beef tissue samples). The isolates were collected by dilution plating on MacConkey agar with sorbitol, cefixime, and tellurite and analyzed by multiplex polymerase chain reaction for virulence genes. Serotyping of non-O157 was performed, and testing for 12 antibiotics by disk diffusion was carried out. A total of 18 STEC strains were isolated, presenting different virulence profiles. Contamination by STEC was observed in the rectum (5/18), carcass surface (5/18), hide (3/18), diaphragm (2/18), retail beef (2/18), and masseter muscle (1/18). Pen floor swabs and esophagus tissues showed no STEC contamination. Moreover, three strains were identified as O26 and three as O113:H21 strains, which have been linked to HUS and HC outbreak cases in Brazil. All STEC isolates were susceptible to all evaluated antimicrobials, except streptomycin. The presence of STEC strains is a direct risk to the consumer, especially when isolated from retail beef, and contamination can occur during different slaughter stages. However, antimicrobial resistance profiles did not identify multidrug-resistant strains, limiting potential antimicrobial resistance transmission to other pathogens.