Enumeration of Chemoorganotrophic Carbonyl Sulfide (COS)-degrading Microorganisms by the Most Probable Number Method.

Carbonyl sulfide (COS) is the most abundant sulfur compound in the atmosphere, and, thus, is important in the global sulfur cycle. Soil is a major sink of atmospheric COS and the numerical distribution of soil microorganisms that degrade COS is indispensable for estimating the COS-degrading potential of soil. However, difficulties are associated with counting COS-degrading microorganisms using culture-dependent approaches, such as the most probable number (MPN) method, because of the chemical hydrolysis of COS by water. We herein developed a two-step MPN method for COS-degrading microorganisms: the first step for chemoorganotrophic growth that supported a sufficient number of cells for COS degradation in the second step. Our new MPN analysis of various environmental samples revealed that the cell density of COS-degrading microorganisms in forest soils ranged between 106 and 108 MPN (g dry soil)-1, which was markedly higher than those in volcanic deposit and water samples, and strongly correlated with the rate of COS degradation in environmental samples. Numerically dominant COS degraders that were isolated from the MPN-positive culture were related to bacteria in the orders Bacillales and Actinomycetales. The present results provide numerical evidence for the ubiquity of COS-degrading microbes in natural environments.

Neural Network Model for Thermal Inactivation of Salmonella Typhimurium to Elimination in Ground Chicken: Acquisition of Data by Whole Sample Enrichment, Miniature Most-Probable-Number Method.

Predictive models are valuable tools for assessing food safety. Existing thermal inactivation models for Salmonella and ground chicken do not provide predictions above 71°C, which is below the recommended final cooked temperature of 73.9°C for chicken. They also do not predict when all Salmonella are eliminated without extrapolating beyond the data used to develop them. Thus, a study was undertaken to develop a model for thermal inactivation of Salmonella to elimination in ground chicken at temperatures above those of existing models. Ground chicken thigh portions (0.76 cm3) in microcentrifuge tubes were inoculated with 4.45 ± 0.25 log most probable number (MPN) of a single strain of Salmonella Typhimurium (chicken isolate). They were cooked at 50 to 100°C in 2 or 2.5°C increments in a heating block that simulated two-sided pan frying. A whole sample enrichment, miniature MPN (WSE-mMPN) method was used for enumeration. The lower limit of detection was one Salmonella cell per portion. MPN data were used to develop a multiple-layer feedforward neural network model. Model performance was evaluated using the acceptable prediction zone (APZ) method. The proportion of residuals in an APZ (pAPZ) from -1 log (fail-safe) to 0.5 log (fail-dangerous) was 0.911 (379 of 416) for dependent data and 0.910 (162 of 178) for independent data for interpolation. A pAPZ ≥0.7 indicated that model predictions had acceptable bias and accuracy. There were no local prediction problems because pAPZ for individual thermal inactivation curves ranged from 0.813 to 1.000. Independent data for interpolation satisfied the test data criteria of the APZ method. Thus, the model was successfully validated. Predicted times for a 1-log reduction ranged from 9.6 min at 56°C to 0.71 min at 100°C. Predicted times for elimination ranged from 8.6 min at 60°C to 1.4 min at 100°C. The model will be a valuable new tool for predicting and managing this important risk to public health.

Comparación de los métodos PetrifilmTM coliformes y Número Más Probable (NMP) para la determinación de coliformes fecales en muestras de queso blanco / Comparison study between the coliform PetrifilmTM and the Most Probable Number methods for the assessment of fecal coliforms in white cheese

Los coliformes fecales son un grupo importante de microorganismos indicadores de inocuidad en alimentos, constituido principalmente por Escherichia coli, el cual es considerado como indicador de contaminación reciente de origen fecal, por ello la importancia de investigar su presencia y determinar rápidamente el nivel poblacional en alimentos. Este trabajo tiene como objetivo comparar el método tradicional Número Más Probable (NMP) para la determinación de coliformes fecales, según Norma Venezolana Covenin Nº 1104-96 con el método de placas rehidratables PetrifilmTM 3M coliformes incubadas en baño de agua circulante (PBA) a 45 ± 0,2°C por 24 ± 2 horas y en estufa con aire circulante 44 ± 1°C por 24 ± 2 horas (PES) de acuerdo con lo recomendado por la Asociación Francesa de Normalización (AFNOR) y la Corporación 3M. Se analizaron un total de 42 muestras de queso blanco aplicando ambas metodologías, dispensando simultáneamente diluciones seriadas de las muestras. Para el análisis estadístico se realizó una correlación de muestras relacionadas (SPS versión 7,5), obteniéndose los siguientes resultados r = 0,952 entre NMP y PES y r = 0,944 entre NMP y PBA; lo que indica una buena correlación positiva entre ambas metodologías en sus diferentes modalidades para la determinación de coliformes fecales en muestras de queso blanco. Se concluye que las placas PetrifilmTM coliformes incubadas a la temperatura óptima de crecimiento de dichos microorganismos es un método alternativo, rápido y confiable para la determinación del nivel de coliformes fecales en queso blanco.

Fecal coliform belong to an important group of sanitary quality indicator microorganisms in food, mainly constituted for Escherichia coli, considerated as indicators of recent fecal contamination, that is why it is very important to investigate their presence and to detect their population in food rapidly. The objetive of this study was to compare the Most Probable Number (NPM) method for fecal coliform determination, according to Venezuelan standard COVENIN Nº 1104-96, with the coliform PetrifilmTM 3M TM plates, incubated in circulating thermostatically - controlled water bath (PBA) at 45 ± 0,2 ° C for 24 ± 2 hours and in a circulating air incubator at 44 ± 1° C for 24 ± 2 hours (PES), according to the recommendation of Association Francoise of Normalization, Paris (AFNOR) and 3M TM corporation. Forty-two white cheese samples were analyzed using both methods mentioned above. They were dispensed at the same time with decimal dilutions of the samples. Data generated were subjected to correlation of related samples (SPS 7.5 version) and the correlation coefficients (r) were obtained; r = 0.952 NMP and PES; r = 0.944 NMP and PBA. It is interesting to observe a good correlation between the methodologies in their different forms for fecal coliform determination in white cheese. Coliform PetrifilmTM plates incubated at the optimal temperature of coliform fecal culture represent a rapid alternative and reliable method for the assessment of fecal coliform population in white cheese.