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Vegetables, especially those eaten raw, have been implicated in several foodborne disease outbreaks. Since multiple vegetable matrices and hazards are involved, risk managers have to prioritize those with the greatest impact on public health to design control strategies. In this study, a scientific-based risk ranking of foodborne pathogens transmitted by leafy green vegetables in Argentina was performed. The prioritization process included hazard identification, evaluation criteria identification and definition, criteria weighting, expert survey design and selection and call for experts, hazard score calculation, hazard ranking and variation coefficient, and result analysis. Regression tree analysis determined four risk clusters: high (Cryptosporidum spp., Toxoplasma gondii, Norovirus), moderate (Giardia spp., Listeria spp., Shigella sonnei), low (Shiga toxin-producing Escherichia coli, Ascaris spp., Entamoeba histolytica, Salmonella spp., Rotavirus, Enterovirus) and very low (Campylobacter jejuni, hepatitis A virus and Yersinia pseudotuberculosis). Diseases caused by Norovirus, Cryptosporidium spp. and T. gondii do not require mandatory notification. Neither viruses nor parasites are included as microbiological criteria for foodstuff. The lack of outbreak studies did not allow to accurately identify vegetables as a source of Norovirus disease. Information on listeriosis cases or outbreaks due to vegetable consumption was not available. Shigella spp. was the main responsible for bacterial diarrhea, but it has not been epidemiologically associated with vegetable consumption. The quality of the available information for all hazards studied was very low and low. The implementation of good practice guidelines throughout the entire vegetable production chain could prevent the presence of the identified hazards. The current study allowed the identification of vacancy areas and could help reinforce the need for performing epidemiological studies on foodborne diseases potentially associated with vegetable consumption in Argentina.

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Abstract We subtyped 32 Salmonella enterica strains isolated from carcasses (n = 10), theenvironment (n = 14), head meat (n = 1) and viscera washing and chilling water (n = 7) in provin-cial abattoirs with no Hazard Analysis Critical Control Point (HACCP) system from Buenos Aires,Argentina, before and after implementing improvement actions. Pulsed-field gel electrophore-sis (PFGE) was carried out using the XbaI restriction enzyme. Strains belonged to six serovars,from which 10 restriction patterns were obtained (five unique patterns and five clusters). Wefound different clones of S. enterica serovars in the same abattoir by XbaI-PFGE. In addition topromoting good hygiene practices, the implementation of an HACCP plan is necessary to meetthe zero-tolerance criteria for Salmonella on beef.

Resumen Subtipificamos en total 32 cepas de Salmonella enterica aisladas de carcasas(n = 10), medio ambiente (n = 14), carne de cabeza (n = 1) y agua de lavado y enfriamientode vísceras (n = 7) en frigoríficos provinciales de Buenos Aires (Argentina) sin análisis de peli-gros y puntos críticos de control (hazard analysis critical control point [HACCP]); la toma demuestras se efectuó antes y después de implementar acciones de mejora. Se llevó a cabo elec-troforesis en gel de campo pulsado (PFGE) utilizando la enzima de restricción XbaI. Las cepaspertenecían a 6 serovares y presentaron 10 patrones de restricción (5 patrones únicos y 5 clus-ters). Demostramos la presencia de diferentes serovares de S. enterica en un mismo frigorífico.

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We subtyped 32 Salmonella enterica strains isolated from carcasses (n=10), the environment (n=14), head meat (n=1) and viscera washing and chilling water (n=7) in provincial abattoirs with no Hazard Analysis Critical Control Point (HACCP) system from Buenos Aires, Argentina, before and after implementing improvement actions. Pulsed-field gel electrophoresis (PFGE) was carried out using the XbaI restriction enzyme. Strains belonged to six serovars, from which 10 restriction patterns were obtained (five unique patterns and five clusters). We found different clones of S. enterica serovars in the same abattoir by XbaI-PFGE. In addition to promoting good hygiene practices, the implementation of an HACCP plan is necessary to meet the zero-tolerance criteria for Salmonella on beef.

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The objective of this study was to develop a quantitative microbial risk assessment (QMRA) model to evaluate potential risk mitigation strategies to reduce the probability of acquiring hemolytic uremic syndrome (HUS) associated with beef consumption in Argentina. Five scenarios were simulated to evaluate the effect of interventions on the probability of acquiring HUS from Shiga toxin-producing Escherichia coli (STEC)-contaminated ground beef and commercial hamburger consumption. These control strategies were chosen based on previous results of the sensitivity analysis of a baseline QMRA model. The application of improvement actions in abattoirs not applying Hazard Analysis and Critical Control Points (HACCP) for STEC would result 7.6 times lower in the probability that consumers acquired HUS from ground beef consumption, while the implementation of improvements in butcher shops would lead to a smaller reduction. In abattoirs applying HACCP for STEC, the risk of acquiring HUS from commercial hamburger consumption was significantly reduced. Treatment with 2% lactic acid, hot water and irradiation reduced 4.5, 3.5 and 93.1 times the risk of HUS, respectively. The most efficient interventions, in terms of case reduction, being those that are applied in the initial stages of the meat chain.

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We developed a quantitative microbiological risk assessment (QMRA) of haemolytic uremic syndrome (HUS) associated with Shiga toxin-producing Escherichia coli (STEC)-contaminated beef (intact beef cuts, ground beef and commercial hamburgers) in children under 15 years of age from Argentina. The QMRA was used to characterize STEC prevalence and concentration levels in each product through the Argentinean beef supply chain, including cattle primary production, cattle transport, processing and storage in the abattoir, retail and home preparation, and consumption. Median HUS probability from beef cut, ground beef and commercial hamburger consumption was <10-15, 5.4x10-8 and 3.5x10-8, respectively. The expected average annual number of HUS cases was 0, 28 and 4, respectively. Risk of infection and HUS probability were sensitive to the type of abattoir, the application or not of Hazard Analysis and Critical Control Points (HACCP) for STEC (HACCP-STEC), stx prevalence in carcasses and trimmings, storage conditions from the abattoir to retailers and home, the joint consumption of salads and beef products, and cooking preference. The QMRA results showed that the probability of HUS was higher if beef cuts (1.7x) and ground beef (1.2x) were from carcasses provided by abattoirs not applying HACCP-STEC. Thus, the use of a single sanitary standard that included the application of HACCP-STEC in all Argentinean abattoirs would greatly reduce HUS incidence. The average number of annual HUS cases estimated by the QMRA (n = 32) would explain about 10.0% of cases in children under 15 years per year in Argentina. Since other routes of contamination can be involved, including those not related to food, further research on the beef production chain, other food chains, person-to-person transmission and outbreak studies should be conducted to reduce the impact of HUS on the child population of Argentina.

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We characterized Shiga toxin-producing Escherichia coli (STEC) O157 (n = 20) and non-O157 (n = 68) isolated from carcasses (n = 54), the environment (n = 20), head meat (n = 3) and viscera washing and chilling water (n = 11) in provincial abattoirs before and after implementing improvement actions. The strains were tested for eae, saa, ehxA and fliCH7 genes. Variants stx1 and stx2 were also determined. Pulsed-field gel electrophoresis (PFGE) was carried out with restriction enzymes XbaI and BlnI. All twenty O157 STEC strains [H7; H21; HNM] carried genes rfbO157 and ehxA; 90.0 % were positive for eae and 15.0 % were negative for fliCH7 and positive for saa. Results of PFGE showed 17 XbaI patterns, of which 14 were unique and three formed clusters. From the 68 non-O157 STEC strains, 66.2 %, 55.9 % and 2.9 % were positive for ehxA, saa and eae genes, respectively. Fifty-three XbaI patterns were obtained (49 unique and four forming clusters). Cross-contamination between products and between the environment and products was confirmed in all abattoirs. While the proposed improvements reduced the risk of contamination, Good Hygiene Practices and Good Manufacturing Practices should be implemented in provincial abattoirs, stressing the importance of having a uniform national food safety standard.

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Abstract We studied and compared the prevalence of Campylobacter jejuni and Campylobacter coli in chicken carcasses from conventional and kosher broiler abattoirs and retail stores. The prevalence of thermotolerant Campylobacter-positive carcasses was 94.0 (kosher) and 32.0% (conventional) (p< 0.0001), while the prevalence of samples contaminated with C. jejuni, C. coli and simultaneously with both species was 36.0, 2.0 and 56.0% (kosher) and 26.0, 4.0 and 2.0% (conventional) (p< 0.0001), respectively. Samples of chicken carcasses (n = 25) and food contact surfaces (tables, n = 25; knives, n=25) from 25 retails were collected and risk quantification was performed. Retails were categorized as high-risk (n = 11), moderate-risk (n = 11) and low-risk (n = 3). Nineteen (76.0%) carcasses, 20 (80.0%) tables and 18 (72.0%) knives were Campylobacter-positive. Retails and abattoirs proved to be sources of carcass contaminaron with Campylobacter spp. Carcasses from kosher abattoirs were mostly contaminated with Campylobacter spp., whereas C. coli was the most prevalent species isolated from carcasses in retail stores.

Resumen El objetivo del estudio fue determinar y comparar la prevalencia de Campylobacter jejuni y Campylobacter coli en carcasas de pollo obtenidas en frigoríficos por faena convencional y kosher, y en locales de expendio. La prevalencia de Campylobacter spp. termotolerante fue del 94,0 (kosher) y del 32,0% (convencional) (p< 0,0001). La prevalencia de muestras contaminadas con C. jejuni, C. coli y con ambas especies fue del 36,0, del 2,0 y del 56,0% (Kosher) y del 26,0, del 4,0 y del 2,0% (convencional) (p< 0,0001), respectivamente. Se tomaron muestras de carcasas (n = 25) y superficies (tablas, n = 25; cuchilla, n = 25) en 25 locales. Los locales fueron categorizados como de riesgo alto (n = 11), moderado (n = 11) y bajo (n = 3). Diecinueve (76,0%) carcasas, 20 (80,0%) tablas y 18 (72,0%) cuchillas fueron positivas para Campylobacter spp. Frigoríficos y locales fueron fuente de contaminación de carcasas con Campylobacter spp. La prevalencia de Campylobacter spp. fue mayor en carcasas kosher. Campylobacter coli fue la especie más prevalente en carcasas de locales.

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The slaughter process plays an important role in animal welfare, meat quality, safety and public health through the meat production chain. In this study, we performed a three-stage evaluation: I) comprehensive evaluation, II) implementation of improvement actions and III) verification of the success of the actions implemented in three abattoirs from Argentina during 2016-2018. Risk was estimated using two checklists, quantified on a 1-100 scale and classified as high (1-40), moderate (41-70) and low (71-100). In stages I and III, Salmonella spp., E. coli O157:H7 and non-O157 STEC were detected and isolated in samples from carcasses (n = 252), the environment (n = 252); head meat (n = 21) and viscera washing and chilling water (n = 105). Carcass samples were analyzed for mesophilic aerobic organisms, coliforms and E. coli enumeration. Of 201 water samples taken, 42.0-75.6 % were non-potable quality. After the implementation of improvement actions in stage II (building, processes, systems for water purification and training), the estimation of risk of contamination was reduced from high to moderate in all three abattoirs, the count of indicator microorganisms decreased in two abattoirs, and the presence of pathogens significantly decreased. Salmonella spp. was not isolated from any of the samples collected in two abattoirs. Isolation of E. coli O157:H7 decreased in carcass and was not isolated from viscera washing and chilling water. Isolation of non-O157 STEC decreased in carcass but not in environmental samples. Finally, 75.0-95.0 % of water samples were of potable quality. Although this was only the first step in the process of change and improvement of abattoirs, the assessment of the situation and the proposal of solutions to correct deviations in a joint effort with the health authorities helped to implement a work model for enhancing food safety before meat reaches consumers.

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We studied and compared the prevalence of Campylobacter jejuni and Campylobacter coli in chicken carcasses from conventional and kosher broiler abattoirs and retail stores. The prevalence of thermotolerant Campylobacter-positive carcasses was 94.0 (kosher) and 32.0% (conventional) (p<0.0001), while the prevalence of samples contaminated with C. jejuni, C. coli and simultaneously with both species was 36.0, 2.0 and 56.0% (kosher) and 26.0, 4.0 and 2.0% (conventional) (p<0.0001), respectively. Samples of chicken carcasses (n=25) and food contact surfaces (tables, n=25; knives, n=25) from 25 retails were collected and risk quantification was performed. Retails were categorized as high-risk (n=11), moderate-risk (n=11) and low-risk (n=3). Nineteen (76.0%) carcasses, 20 (80.0%) tables and 18 (72.0%) knives were Campylobacter-positive. Retails and abattoirs proved to be sources of carcass contamination with Campylobacter spp. Carcasses from kosher abattoirs were mostly contaminated with Campylobacter spp., whereas C. coli was the most prevalent species isolated from carcasses in retail stores.

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Antimicrobial treatments could help to decrease the transmission of microorganisms to beef carcasses and abattoir environments. The aim of this study was to evaluate the effectiveness of interventions in reducing Shiga toxin genes (stx1 and stx2) presence in a commercial abattoir. Intervention measures included the application of electrolytically generated hypochlorous acid to steer pens (experiment 1), chlorinated water, electrolytically generated hypochlorous acid, and isoclor to steer pens (experiment 2), electrolytically generated hypochlorous acid to knocking pens (experiment 3), and aqueous ozone and electrolytically generated hypochlorous acid onto beef carcasses (experiment 4). Detection of stx in samples was performed with BAX® System Real-Time PCR Assay. Our results showed that treatment with pressurized electrolytically generated hypochlorous acid and isoclor were effective to reduce stx presence from hides on steer pens. Although there is no single strategy to ensure the reduction of stx presence in a commercial abattoir, the combined application of several antimicrobial interventions would be ideal.

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