Comprehensive evaluation of abattoirs with no Hazard Analysis Critical Control Point plan in Tucumán,Argentina / Evaluación integral de frigoríficos sin un plan de análisis de peligros y puntos críticos de control en Tucumán, Argentina

Abstract This work focused on the comprehensive study of two provincial transit abattoirs inTucumán, Argentina, with no Hazard Analysis Critical Control Point (HACCP) plan. Visits (n = 20)were conducted between 2016 and 2018 during the operational and post-operational processes.Risk was estimated and the bacteriological analysis of carcass and environmental samples wasperformed. Risk estimation showed the predominance of high risk in both abattoirs. The maindeviations from the HACCP plan were: deficient building conditions, deficient workflow, lack of sectorization of changing rooms and bathrooms, lack of implementation of Standardized Sanitary Operational Procedures, and no food safety training of workers. The counts of indi-cator microorganisms from both abattoirs were not significant. Salmonella spp. was isolated from 7.5% carcass and 7.3% environmental samples. The Salmonella serovars identified were Cerro, Corvallis, Havana and Agona. Shiga toxin (stx) genes were detected in 24.4% carcass and 30.9% environmental samples. The isolates were characterized as Escherichia coli O8:H7/stx1, O116:H49/stx2 and O136:H40/stx2. Based on these results, it would be possible to implement an improvement plan in Tucumán abattoirs together with the local health authorities. Still, the need to work jointly with the sanitary authority in search of a unique sanitary standard for Argentina remains unaddressed.

Resumen Este trabajo se centró en el estudio integral de dos frigoríficos de tránsito provincial en Tucumán, Argentina, carentes de un plan de análisis de peligros y puntos críticos de control (HACCP, por sus siglas en inglés). Las visitas (n = 20) se realizaron entre 2016 y 2018 durante los procesos operativos y posoperativos. Se realizó la estimación del riesgo y el análisis bacteriológico de medias reses y muestras ambientales. La estimación del riesgo demostró un predominio de riesgo alto en ambos frigoríficos. Las principales desviaciones del plan HACCP fueron las deficientes condiciones edilicias, un inadecuado flujo de trabajo, la falta de sectorización de vestuarios y banños, una implementación nula de procedimientos operativos estandarizados de saneamiento y una insuficiente capacitación en seguridad alimentaria de los operarios. Los recuentos de microorganismos indicadores de ambos frigoríficos no presentaron diferencias significativas. Salmonella spp. se aisló del 7,5% de muestras de medias reses y del 7,3% de muestras ambientales. Se identificaron las siguientes serovariedades de Salmonella: Cerro, Corvallis, Havana y Agona. Se detectaron genes de toxina Shiga (sfx) en el 24,4% de las muestras de medias reses y en el 30,9% de las muestras ambientales. Los aislamientos se caracterizaron como Escherichia coli O8:H7/sfx1, O116:H49/sfx2 y O136:H40/sfx2. Teniendo en cuenta estos resultados, sería posible implementar un plan de mejoramiento en frigoríficos de Tucumán conjuntamente con las autoridades locales de salud. Aun así, sigue sin abordarse la necesidad de trabajar en vinculación con las autoridades sanitarias en la búsqueda de una norma integrada única para Argentina.

Molecular subtyping of Salmonella spp. strains in provincial abattoirs with no hazard analysis critical control point from Buenos Aires, Argentina / Subtipificación molecular de Salmonella spp. en frigoríficos provinciales sin análisisde peligros y puntos críticos de control ubicados en Buenos Aires, Argentina

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.

Comparación de tres técnicas para subtipificación molecular de Listeria monocytogenes / Comparison of three molecular subtyping techniques for Listeria monocytogenes

Abstract Listeria monocytogenes is a foodborne pathogen. The recent alert for L. monocytogenes in vegetables from Argentina warns about the importance of reinforcing its isolation, characterization and subtyping in food, clinical and environmental samples. The aim of the present study was to compare the discriminatory power of enterobacterial repetitive interpower; genic consensus polymerase chain reaction (ERIC-PCR), automated ribotyping and pulsed-field gel electrophoresis (PFGE) to subtype strains of L. monocytogenes isolated from Argentine meat and environmental samples. Simpson's Diversity Index (DI) was calculated on the basis of based on the dendrograms obtained in the by cluster analysis, showing the following discriminatory power: ApaI-PFGE (0.980), AscI-PFGE (0.966), ribotyping (0.912), ERIC-PCR (0.886). The ID values between ApaI- and AscI-PFGE and between ribotyping and ERIC-PCR were not significantly different. Of the three techniques evaluated, PFGE showed the highest discriminatory power. However, the subtyping techniques should be accompanied by effective food monitoring strategies and reliable clinical and epidemiological studies.

Resumen Listeria monocytogenes es un patógeno alimentario. La reciente alerta por la presencia de L. monocytogenes en vegetales en Argentina advierte sobre la importancia de reforzar el aislamiento, la caracterización y la subtipificación de esta bacteria en muestras clínicas de alimentos y ambientales. El objetivo del presente estudio fue comparar el poder discriminatorio de enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR), la ribotipificación automatizada y la pulsed-field gel electrophoresis (PFGE) para subtipificar cepas de L. monocytogenes aisladas de carne y de muestras ambientales en Argentina. El índice de diversidad (ID) de Simpson, calculado a partir de los dendrogramas obtenidos en el análisis de agrupamiento, mostró los siguientes resultados: Apal-PFGE (0,980), AscI-PFGE (0,966), riboti-pado (0,912), ERIC-PCR (0,886). Los valores obtenidos no fueron significativamente diferentes al comparar entre Apal- y AscI-PFGE, ni entre ribotipadoy ERIC-PCR. De las técnicas evaluadas, la PFGE presentó el mayor poder discriminatorio. Sin embargo, las técnicas de subtipificación deberían acompañarse de estrategias de control de los alimentos efectivas y de estudios clínicos y epidemiológicos confiables.

Estudos laboratoriais para avaliação do desempenho de camadas de cobertura de aterros sanitários em relação à redução de emissões de gases e infiltrações / Laboratory studies to evaluate the performance of landfill cover layers for the reduction of gases emissions and infiltrations

RESUMO As camadas de cobertura de aterros sanitários são construídas para impedir a saída de gases para a atmosfera e a entrada de líquidos no interior do aterro, minimizando os impactos no meio ambiente. Este trabalho avaliou, por meio de ensaios de colunas de solos em laboratório, o desempenho de duas camadas de cobertura, compreendendo o comportamento dos solos em função das emissões de metano (CH4) e da infiltração de água em seu interior. Foram utilizadas duas configurações de camada de solos: Coluna 1, camada convencional com solo com espessura 0,60 m e grau de compactação de 80%; e Coluna 2, camada oxidativa, com espessura de 0,30 m de solo com grau de compactação de 80%, sobreposta por uma camada de 0,30 m de solo misturado com produto compostado na proporção de 1:1 em volume com grau de compactação de 76%. Após a confecção das colunas (dimensões úteis de 0,60 m de altura e 0,15 m de diâmetro), foi injetado CH4 em sua parte inferior com concentração de 100% na vazão de 0,5 L/h (8,3 mL/min ou 486 g/m2.dia) para se avaliar a redução das emissões. Posteriormente, foi simulada chuva de 40 mm - sendo 30 mm em 1 dia e 10 mm 3 dias após a primeira simulação - e avaliada a infiltração da água e as emissões de CH4 nas colunas. Os resultados obtidos indicam que as duas configurações de camadas de cobertura apresentaram resultados satisfatórios, diminuindo o volume de líquidos para o interior do aterro e minimizando as emissões de CH4 para a atmosfera. Observou-se que a Coluna 2 (0,30 m de solo + 0,30 m de solo + produto compostado) se mostrou mais eficiente tanto na redução de emissões como na capacidade de retenção de água em relação à Coluna 1 (0,60 m de solo), sugerindo que camadas oxidativas podem se tornar uma alternativa tecnológica para regiões onde exista escassez de solos argilosos.

ABSTRACT Landfill cover layers are built to prevent the release of gases into the atmosphere and the inflow of liquids into the landfill, minimizing impacts on the environment. This study evaluated, through the testing of soil columns in the laboratory, the performance of two cover layers, comprehending the behavior of the soils as a function of methane emissions and liquid flows in its interior. Two configurations were used in the soil layer: Column 1, conventional layer with compacted soil of 0.60 m thickness and 80% degree of compaction; and Column 2, oxidative layer with 0.30 m thickness of soil and 80% degree of compaction, stuck by a layer of 0.30 m of mixed soil with composted product in the volume ratio of 1:1 with 76% degree of compaction. After the confection of the columns (0.60 m in height and 0.15 m in diameter), methane gas was injected into the inferior part of the columns with a concentration of 100% at a flow rate of 0.5 L/h (8.3 mL/min or 486 g/m2.day) to evaluate the emission reduction. Subsequently, a rain of 40 mm was simulated - being 30 mm in one day and 10 mm three days after the first simulation -, and both the methane emissions and the infiltration of water in the columns were evaluated. The obtained results indicate that the two column configurations of landfill cover layers presented satisfactory results, reducing the volume of liquids into the landfill and minimizing the methane emissions into the atmosphere. It is observed that Column 2 (0.30 m of soil + 0.30 m of soil + composted product) was more efficient in the reduction of emissions, as well as in the retention capacity of water in relation to Column 1 (0.60 m of soil), suggesting that oxidative layers can become a technological alternative for regions where there is scarcity of clay soils.