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.

Influence of temperature and pH on induction of Shiga toxin Stx1a in Escherichia coli.

Shiga toxin-producing strains represent pathogenic group that is of concern in food production. The present study evaluated forty-eight E. coli isolates (11 with intact stx gene, while remaining isolates presented only stx-fragments) for Shiga toxin production. The four most expressive stx-producers (O26, O103, O145, and O157) were selected to evaluate effects of pH (3.5, 4.5, and 7) and temperature (35, 40, and 50°C). After determining acid stress effects in media on Stx-induction, we mimicked "in natura" conditions using milk, apple, and orange juices. Only isolates that showed the presence of intact stx gene (11/48) produced Shiga toxin. In addition, acid pH had a role in down-regulating the production of Shiga toxin, in both lactic acid and juices. In contrast, non-lethal heating (40°C), when in neutral pH and milk was a favorable environment to induce Shiga toxin. Lastly, two isolates (O26 and O103) showed a higher capacity to produce Shiga toxin and were included in a genomic cluster with other E. coli involved in worldwide foodborne outbreaks. The induction of this toxin when subjected to 40°C may represent a potential risk to the consumer, since the pathogenic effect of oral ingestion of Shiga toxin has already been proved in an animal model.

Efficacy of Quaternary Ammonium Compounds for Control of Individual and Mixed Cultures of Escherichia coli with High- and Low-Quaternary Ammonium Compounds Resistance.

Escherichia coli is a well-characterized micro-organism in scientific literature. Similarly, quaternary ammonium compounds (QACs) are historical sanitizers in food processing. However, the use of QACs has been questioned due to bacterial resistance in some studies. Therefore, this study aimed to compare effects of single and mixed cultures of E. coli strains of different serogroups with either high (six strains) or low (five strains) resistance to QACs. Twenty-five combinations of strains with either high (H)- or low (L)-QAC resistance were analyzed (H + H vs. L + L). After exposure to QAC, combinations with statistical differences (p < 0.05) compared with individuals were selected and an inactivation model determined using GInaFit®. Only one combination of two strains (C23 and C20) with low-QAC resistance (mixture T18) had greater resistance (p < 0.05) than the individual isolates. The combination T18 and individual strain C23 presented a Weibull model, whereas the other isolated strain (C20) presented a biphasic inactivation model with a shoulder. Whole genome sequencing determined that unlike C20, C23 carried yehW, which may have led to Weibull inactivation. Possibly, very rapid interaction of C20 with the QAC favored increased survival of C23 and overall persistence of the T18 mixture. Consequently, our results indicate that individual E. coli with low-QAC resistance can synergistically interfere with QAC inactivation.

Phenolic Compounds in Bacterial Inactivation: A Perspective from Brazil.

Phenolic compounds are natural substances that are produced through the secondary metabolism of plants, fungi, and bacteria, in addition to being produced by chemical synthesis. These compounds have anti-inflammatory, antioxidant, and antimicrobial properties, among others. In this way, Brazil represents one of the most promising countries regarding phenolic compounds since it has a heterogeneous flora, with the presence of six distinct biomes (Cerrado, Amazon, Atlantic Forest, Caatinga, Pantanal, and Pampa). Recently, several studies have pointed to an era of antimicrobial resistance due to the unrestricted and large-scale use of antibiotics, which led to the emergence of some survival mechanisms of bacteria to these compounds. Therefore, the use of natural substances with antimicrobial action can help combat these resistant pathogens and represent a natural alternative that may be useful in animal nutrition for direct application in food and can be used in human nutrition to promote health. Therefore, this study aimed to (i) evaluate the phenolic compounds with antimicrobial properties isolated from plants present in Brazil, (ii) discuss the compounds across different classes (flavonoids, xanthones, coumarins, phenolic acids, and others), and (iii) address the structure-activity relationship of phenolic compounds that lead to antimicrobial action.

Association of resistance to quaternary ammonium compounds and organic acids with genetic markers and their relationship to Escherichia coli serogroup.

Sanitizer resistance is being extensively investigated due to the potential for bacterial survival and cross-resistance with other antimicrobials. Similarly, organic acids are being used due to their microbial inactivation potential as well as being generally recognized as safe (GRAS). However, little is known about associations of genetic and phenotypic factors in Escherichia coli related to resistance to sanitizers and organic acids as well as differences between "Top 7" serogroups. Therefore, we investigated 746 E. coli isolates for resistance to lactic acid and two commercial sanitizers based on quaternary ammonium and peracetic acid. Furthermore, we correlated resistance to several genetic markers and investigated 44 isolates using Whole Genome Sequencing. Results indicate that factors related to motility, biofilm formation, and Locus of Heat Resistance played a role in resistance to sanitizers and lactic acid. In addition, Top 7 serogroups significantly differed in sanitizer and acid resistance, with O157 being the most consistently resistant to all treatments. Finally, mutations in rpoA, rpoC, and rpoS genes were observed, in addition to presence of a Gad gene with alpha-toxin formation in all O121 and O145 isolates, which may be related to increased resistance of these serogroups to the acids used in the present study.

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.

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.

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.

Salmonella in the processing line of farmed Tambatinga (Colossoma macropomum x Piaractus brachypomus) in Mato Grosso, Brazil: serotypes of occurrence and antimicrobial profile.

The objective of this study was to evaluate the dispersion dynamics and antimicrobial resistance profiles of Salmonella in the processing of Tambatinga (Colossoma macropomum x Piaractus brachypomus). Thirty fish were monitored during four processing stages (reception, first wash, evisceration, and prepackage area) in a fish slaughterhouse. One hundred and twenty fish surface samples were collected and tested through bacteriological analysis, PCR, serotyping, and antimicrobial resistance profile (disk-diffusion). Of these samples, 7.5% (9/120) were positive for Salmonella, with 0.83% being observed in the pre-packaging phase, indicating a low occurrence at this stage. All the analyzed stages were positive for Salmonella, with the prevalent serovars being Ndolo, Mbandaka, Typhimurium, Rough, and O:16. All strains were sensitive to various antimicrobials. Improvements in microbiological control during all processing stages should be implemented to ensure a Salmonella-free product.

Salmonella Schwarzengrund, Akuafo, and O:16 isolated from vacuum-packaged beef produced in the state of Mato Grosso, Brazil.

INTRODUCTION: Salmonella spp. is a pathogen associated with foodborne infections, mainly in foods of animal origin. In this context, the present study investigated the occurrence of Salmonella serotypes, genotypes and the antimicrobial resistance profiles of strains in fresh beef produced in Mato Grosso, Brazil. METHODOLOGY: A total of 107 samples from 13 different slaughterhouses in the Mato Grosso were analyzed. Suggestive Salmonella spp. colonies detected during the biochemical screening were submitted to DNA extraction, and hilA gene amplification was used for the PCR reaction. Antimicrobial resistance analyses were performed using 17 antimicrobial agents from eight different classes by the disk diffusion method. Strains exhibiting multiple drug resistances were submitted to PCR genotyping based on repetitive sequences (rep-PCR), using a commercial semiautomatic DiversiLab® system. RESULTS: A total of 5.6% (6/107) of the samples tested positive by the conventional method and were confirmed by PCR, namely two S. Akuafo, two non-typable Salmonella enterica strains, one Salmonella O:16 serovar, and one S. Schwarzengrund. The antimicrobial resistance profiles indicated resistance to gentamicin (30%), tetracycline, nitrofurantoin, and trimethoprim + sulfamethoxazole (16%). Genotyping indicated a 70% difference between S. Schwarzengrund and the non-typable Salmonella strains. No genetic similarities were observed between the six Salmonella isolates based on rep-PCR, including two S. Akuafo. CONCLUSIONS: The results obtained herein corroborate that Salmonella serovar Schwarzengrund is commonly isolated in animal products in the state of Mato Grosso, Brazil, also highlighting the presence of two unusual Salmonella serovars in beef (Akuafo and O:16).

Everybody loves cheese: crosslink between persistence and virulence of Shiga-toxin Escherichia coli.

General cheese manufacturing involves high temperatures, fermentation and ripening steps that function as hurdles to microbial growth. On the other hand, the application of several different formulations and manufacturing techniques may create a bacterial protective environment. In cheese, the persistent behavior of Shiga toxin-producing Escherichia coli (STEC) relies on complex mechanisms that enable bacteria to respond to stressful conditions found in cheese matrix. In this review, we discuss how STEC manages to survive to high and low temperatures, hyperosmotic conditions, exposure to weak organic acids, and pH decreasing related to cheese manufacturing, the cheese matrix itself and storage. Moreover, we discuss how these stress responses interact with each other by enhancing adaptation and consequently, the persistence of STEC in cheese. Further, we show how virulence genes eae and tir are affected by stress response mechanisms, increasing either cell adherence or virulence factors production, which leads to a selection of more resistant and virulent pathogens in the cheese industry, leading to a public health issue.

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.

Whole-Genome Draft Assemblies of Difficult-to-Classify Escherichia coli O157 and Non-O157 Isolates from Feces of Canadian Feedlot Cattle.

Forty-eight Escherichia coli strains were chosen due to variable detection of stx or serogroup by PCR. Although all strains were initially determined to be Shiga toxin-producing Escherichia coli (STEC), their genomes revealed 11 isolates carrying stx 1a, stx 1b, stx 2a, and/or stx 2b Assembled genome sizes varied between 4,667,418 and 5,556,121 bp, with N 50 values between 79,648 and 294,166 bp and G+C contents between 50.3% and 51.4%.