Assessment of hygiene practices and microbial safety of milk supplied by smallholder farmers to processors in selected counties in Kenya.

Smallholder farmers dominate the Kenyan dairy sector producing 95% of the total milk. However, several concerns have been raised on the quality and safety of the milk they produce. This study assessed the hygienic practices and microbial safety of milk supplied by smallholder farmers to processors in Bomet, Nyeri, and Nakuru counties in Kenya. Interviews and direct observations were carried out to assess hygiene and handling practices by farmers and a total of 92 milk samples were collected along four collection channels: direct suppliers, traders, cooperatives with coolers, and cooperatives without coolers. Microbial analysis was done following standard procedures and data analysed using GenStat and SPSS. This study revealed that farmers did not employ good hygienic practices in their routine dairy management. They used plastic containers for milking and milk storage (34.2%); they did not clean sheds (47.9%) and did not set aside cows that suffered from mastitis factors (83.6%), resulting in poor microbial quality of raw milk along the collection channels. The highest mean total viable counts (8.72 log10 cfu/ml) were recorded in Nakuru while Nyeri had the highest mean E. coli counts (4.97 log10 cfu/ml) and Bomet recorded the highest mean counts of 5.13 and 5.78 log10 cfu/ml for Staphylococcus aureus and Listeria monocytogenes respectively. Based on all above-mentioned parameters, the microbial load in most samples from all three counties exceeded the set Kenyan standards. Farmer training, improving road infrastructure, use of instant coolers at cooperatives, and quality-based payment systems are recommended as measures to curb microbial growth.

To culture or not to culture: careful assessment of metabarcoding data is necessary when evaluating the microbiota of a modified-atmosphere-packaged vegetarian meat alternative throughout its shelf-life period.

BACKGROUND: As the increased consumption of ready-to-eat meat alternatives is a fairly recent trend, little is known about the composition and dynamics of the microbiota present on such products. Such information is nonetheless valuable in view of spoilage and food safety prevention. Even though refrigeration and modified-atmosphere-packaging (MAP) can extend the shelf-life period, microbial spoilage can still occur in these products. In the present study, the microbiota of a vegetarian alternative to poultry-based charcuterie was investigated during storage, contrasting the use of a culture-dependent method to a culture-independent metagenetic method. RESULTS: The former revealed that lactic acid bacteria (LAB) were the most abundant microbial group, specifically at the end of the shelf-life period, whereby Latilactobacillus sakei was the most abundant species. Metabarcoding analysis, in contrast, revealed that DNA of Xanthomonas was most prominently present, which likely was an artifact due to the presence of xanthan gum as an ingredient, followed by Streptococcus and Weissella. CONCLUSIONS: Taken together, these results indicated that Lb. sakei was likely the most prominent specific spoilage organisms (SSO) and, additionally, that the use of metagenetic analysis needs to be interpreted with care in this specific type of product. In order to improve the performance of metagenetics in food samples with a high DNA matrix but a low bacterial DNA load, selective depletion techniques for matrix DNA could be explored.

Assessment of the sanitary quality of ready to eat sesame, a low moisture street food from Burkina Faso.

BACKGROUND: Microbial contamination of edible low moisture food poses a significant public health risk for human. In this study, the microbial quality of sweet dehulled sesame seed croquettes, salted dehulled sesame seed and the raw sesame seed, sold under ambient conditions were examined. The samples were collected in the cities of Burkina Faso. The first type is sweet dehulled sesame seed croquettes (n1 = 25); the second type is salted dehulled sesame seed (n2 = 25) and the third type is raw sesame seed (n3 = 25). Assessment of the microbial quality was based on the total aerobic mesophilic bacteria, the thermotolerant coliforms, the yeasts and moulds, the E. coli, and the Salmonella spp. using ISO methods. RESULTS: The results showed the presence of microorganisms varying from <1.0 to 1.72 × 105 CFU g- 1 for thermotolerant coliforms, from <1.0 to 6,12 × 106 CFU g- 1 for the total mesophilic aerobic flora and from <1.0 to 8.10 × 105 CFU g- 1 for yeasts and moulds. The higher contaminations rates were mostly observed in raw sesame seed samples. No E coli or Salmonella pathogens were detected. Based on international standards of dehydrated food, 50.67% of the ready to eat sesame are satisficing while 17.33% are acceptable and 32% are not satisficing. CONCLUSION: Attention should be emphasized on the processing practices, especially in crowded places where RTE sesames seeds are mostly sold. The high numbers of all microbial groups in these sesame seed samples suggested that the production of RTE sesame seed should be improved by better hygiene. This study highlights also that RTE sesame seed might harbor a wide range of microorganisms when processes are weak of hygiene.

Validation of a High-Throughput Sausage Casing Model for the Assessment of Bacterial Inactivation Affected by Salt Concentration, pH, and Temperature.

Previous studies have shown the efficacy of high concentrations of salt as the main preservative against vegetative bacteria present on natural sausage casings. These studies were limited in the number of variables and the interactions between these variables that were assessed. To remedy this situation, a MicroCasing high-throughput model was developed and validated to study the inactivation kinetics of various combinations of parameters (salt concentration, pH, and temperature) on eight bacterial isolates of Salmonella enterica, Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes over a prolonged period. A Weibullian power model was the best fit to show the trends in sensitivity of each bacterial isolate to salt, pH, and temperature over time. The inactivation kinetics generated with this novel approach could serve as a predictive model for the required salting period for casings. The actual bacterial contamination of the product can vary with the respective production step during processing from animal intestine into sausage casings (initial level, ∼105 CFU/g; level after salting, <102 CFU/g). Subsequent selection and grading of these casings will require complete removal of all salt, and upon completion of this production step, the casings will be resalted. By determining the actual contamination level before the salting process, the minimum storage period in salt can be calculated and potentially optimized by adjusting the pH and temperature. As a result, a standard holding period of at least 30 days may no longer be necessary to produce salted natural casings in accordance with validated quality and food safety criteria.

The qualified presumption of safety assessment and its role in EFSA risk evaluations: 15 years past.

Microorganisms are intentionally added at different stages of the food and feed chain (food or feed additive, novel food or plant protection product) and are subjected to regulation and safety assessment by the European Food Safety Authority. Safety evaluation is based on application dossiers for market authorisation to the European Commission. The qualified presumption of safety (QPS) concept was developed in 20031 to provide a harmonised generic safety pre-appraisal of the above microorganisms. Unambiguously defined biological taxonomic units (TUs) are assessed for their body of knowledge, their safety and their end use. Identified safety concerns for a certain TU can be, where reasonable in number and not universally present, reflected as 'qualifications.' Strains belonging to TUs having QPS status may benefit of a fast track evaluation. The lowest TU for which the QPS status is granted is the species level for bacteria and yeasts and the family for viruses. The QPS concept is also applicable to genetically modified microorganisms used for production purposes. Based on the current body of knowledge and/or the ambiguous taxonomic position, some TUs, such as filamentous fungi, bacteriophages, Enterococcus faecium, Escherichia coli, Streptomyces spp. and Oomycetes, are not considered liable for QPS status.

Next generation microbiological risk assessment-Potential of omics data for hazard characterisation.

According to the World Health Organization estimates in 2015, 600 million people fall ill every year from contaminated food and 420,000 die. Microbial risk assessment (MRA) was developed as a tool to reduce and prevent risks presented by pathogens and/or their toxins. MRA is organized in four steps to analyse information and assist in both designing appropriate control options and implementation of regulatory decisions and programs. Among the four steps, hazard characterisation is performed to establish the probability and severity of a disease outcome, which is determined as function of the dose of toxin and/or pathogen ingested. This dose-response relationship is subject to both variability and uncertainty. The purpose of this review/opinion article is to discuss how Next Generation Omics can impact hazard characterisation and, more precisely, how it can improve our understanding of variability and limit the uncertainty in the dose-response relation. The expansion of omics tools (e.g. genomics, transcriptomics, proteomics and metabolomics) allows for a better understanding of pathogenicity mechanisms and virulence levels of bacterial strains. Detection and identification of virulence genes, comparative genomics, analyses of mRNA and protein levels and the development of biomarkers can help in building a mechanistic dose-response model to predict disease severity. In this respect, systems biology can help to identify critical system characteristics that confer virulence and explain variability between strains. Despite challenges in the integration of omics into risk assessment, some omics methods have already been used by regulatory agencies for hazard identification. Standardized methods, reproducibility and datasets obtained from realistic conditions remain a challenge, and are needed to improve accuracy of hazard characterisation. When these improvements are realized, they will allow the health authorities and government policy makers to prioritize hazards more accurately and thus refine surveillance programs with the collaboration of all stakeholders of the food chain.

Microbiological criteria and ecology of commercially available processed cheeses according to the product specification and physicochemical characteristics.

Although global cheese manufacturers release a variety of products onto the market, research on the microbiological quality and safety of cheese has focused mainly on conventional cheeses made from milk. Here, this study aimed to investigate commercially processed cheese products produced by mixing conventional cheeses after melting. Two approaches were used: a summary and comparison of legal definitions and standards/regulations regarding the microbiological criteria used by major cheese traders in the global market (Australia/New Zealand, China, European Union, Japan, Mexico, Republic of Korea, and the United States) and a comprehensive microbiological analysis of commercial products (n = 800), along with an assessment of salinity, pH, water activity, and heating conditions. The results of the literature search showed that major importing countries (China, Japan, Mexico, and the Republic of Korea) have stricter microbiological criteria for commercially available cheese products than major exporters (Australia/New Zealand, EU, and the USA). The former set limits with respect to the number of total coliforms in the product. Microbiological analyses were designed according to global standards and recommendations. No test sample contained detectable levels of Clostridium perfringens, enterohemorrhagic Escherichia coli, Listeria monocytogenes, Salmonella, or Staphylococcus aureus. In addition, no coliform bacteria (including E. coli) were detected. Overall, 79.9% of the samples contained detectable aerobic plate counts (1.0-7.8 log CFU/g); these levels varied significantly according to product type (grated cheese > chunks; cream cheese > portions or sliced) (p < .05). There was no significant association between microbe levels and salinity, water activity, pH, and heating conditions. The results can be used to develop a comprehensive database about commercially processed cheese products available in the global market and, as such, may be helpful for both national authorities and cheese manufacturers when considering novel strategic management plans for microbiological quality and safety.

The STARTEC Decision Support Tool for Better Tradeoffs between Food Safety, Quality, Nutrition, and Costs in Production of Advanced Ready-to-Eat Foods.

A prototype decision support IT-tool for the food industry was developed in the STARTEC project. Typical processes and decision steps were mapped using real life production scenarios of participating food companies manufacturing complex ready-to-eat foods. Companies looked for a more integrated approach when making food safety decisions that would align with existing HACCP systems. The tool was designed with shelf life assessments and data on safety, quality, and costs, using a pasta salad meal as a case product. The process flow chart was used as starting point, with simulation options at each process step. Key parameters like pH, water activity, costs of ingredients and salaries, and default models for calculations of Listeria monocytogenes, quality scores, and vitamin C, were placed in an interactive database. Customization of the models and settings was possible on the user-interface. The simulation module outputs were provided as detailed curves or categorized as "good"; "sufficient"; or "corrective action needed" based on threshold limit values set by the user. Possible corrective actions were suggested by the system. The tool was tested and approved by end-users based on selected ready-to-eat food products. Compared to other decision support tools, the STARTEC-tool is product-specific and multidisciplinary and includes interpretation and targeted recommendations for end-users.

Regulatory Issues Associated with Preharvest Food Safety: European Union Perspective.

Free movement of safe and wholesome food is an essential aspect of any society. This article contains an updated description of the regulatory issues associated with preharvest food safety within the European Union. Salmonella, Campylobacter, Trichinella, antimicrobial resistance, and bovine spongiform encephalopathy are dealt with in detail. Moreover, Cysticercus bovis/Taenia saginata, Toxoplasma, Yersinia, verotoxigenic/shigatoxigenic Escherichia coli, Listeria, and foodborne viruses are briefly covered. The article describes how the focus in the European Union is changing to involve a supply chain view with a focus on cost-effectiveness. The precautionary principle-as well as the use of private standards as an instrument to ensure compliance-is dealt with. In addition, actions in the pipeline are presented and discussed.

Integrated assessment of biomarker responses and microbiological analysis of oysters from São Luís Island, Brazil.

This study was conducted to evaluate the use of biochemical biomarkers and microbiological analysis to identify levels of oyster contamination at different ports in São Luís Island (Maranhão), Brazil. Oysters were analyzed for total coliforms, thermotolerant coliforms, Escherichia coli and Aeromonas spp. In addition, tissue was removed from the digestive gland to determine the glutathione-S-transferase (GST) and catalase (CAT) activity. The highest percentage of microbiological contamination of oyster samples occurred during the rainy season. The activity of GST and catalase in oysters was also higher in the rainy season, coinciding with the greatest abundance of total and thermotolerant coliforms. Among the prospective biomarkers, GST showed the best results for identification of areas with higher levels of contamination.

Food hygiene assessment in catering establishments in Hay Hassani district-Casablanca.

Contaminated food is responsible for a significant amount of illnesses. In Morocco, it has become a worrying concern. Numerous awareness campaigns are conducted to warn the population against the risks of such scourge in ways that will prevent foodborne illness. Lawful commissions are in charge of examining and ensuring food safety in production and catering establishments, in addition to the assessment of food poisoning risks. The aim of this study is to evaluate the hygienic quality of food handling, preparation, and storage in catering establishments within Hay Hassani prefecture in Casablanca. During the period 2006-2012 a total of 1765 food samples were taken and examined for microbiological quality tests. As analyzed, 562 per 1765 samples are declared unhealthy for consumption. We note that some products were highly contaminated as compared to other products (p <0.001), specifically vegetable dishes, and meat dishes. In Hay Hassani district food is generally prepared and sold under unhygienic conditions, adequate corrective measures have been announced to improve hygienic practices.

[Evaluation of the presence of hygienic and sanitary indicator microorganisms in food served in public schools in Porto Alegre, Brazil]. / Avaliação da presença de microrganismos indicadores higiênico-sanitários em alimentos servidos em escolas públicas de Porto Alegre, Brasil.

The objective of this study was to evaluate the presence of hygienic and sanitary indicator microorganisms in samples of food served in public schools in Porto Alegre. All the food served in the meal of the session visited was analyzed for Escherichia coli, coagulase-positive Staphylococcus, Salmonella sp. and Shigella sp. Of the total of 196 food products analyzed in 120 schools, 4 contained and Escherichia coli score above the permitted level, and 2 contained coagulase-positive Staphylococcus. Neither Shigella nor Salmonella genus were detected. In the majority of schools studied, it was found that food was of an adequate hygienic-sanitary standard. However, only municipal schools had the supervision of a technician responsible for school food. In the state schools, 60% had never been visited by a nutritionist and in these schools several procedures failed to comply with legal requirements. In most of the schools studied, the food served to students was within adequate standards, though the problems detected revealed the need for the implementation of Best Practices in the school environment.

Avaliação da presença de microrganismos indicadores higiênico-sanitários em alimentos servidos em escolas públicas de Porto Alegre, Brasil / Evaluation of the presence of hygienic and sanitary indicator microorganisms in food served in public schools in Porto Alegre, Brazil

Este estudo tem por objetivo avaliar a presença de microrganismos indicadores higiênico-sanitários em amostras de alimentos servidos em escolas públicas de Porto Alegre. Foram analisados todos os alimentos servidos na refeição do turno da visita, quanto à presença de Escherichia coli, Staphylococcus coagulase positiva, Salmonella sp. e Shigella sp. No total de 196 alimentos analisados de 120 escolas, 4 apresentavam contagem de Escherichia coli acima do permitido e dois tinham a presença de Staphylococcus coagulase positiva. Os gêneros Shigella e Salmonella não foram encontrados. Foi observado que a maioria das escolas estudadas servia alimentos dentro de padrões higiênico-sanitários adequados. Foi evidenciado que somente escolas municipais contavam com a orientação de responsável técnico pela alimentação escolar. Das escolas estaduais 60% nunca haviam recebido visita de nutricionista nas quais foram encontrados procedimentos em desacordo com as exigências da legislação. Na maioria das escolas, os alimentos servidos estavam dentro de padrões adequados, porém os problemas detectados demonstram a necessidade da implantação das Boas Práticas no ambiente escolar.

The objective of this study was to evaluate the presence of hygienic and sanitary indicator microorganisms in samples of food served in public schools in Porto Alegre. All the food served in the meal of the session visited was analyzed for Escherichia coli, coagulase-positive Staphylococcus, Salmonella sp. and Shigella sp. Of the total of 196 food products analyzed in 120 schools, 4 contained and Escherichia coli score above the permitted level, and 2 contained coagulase-positive Staphylococcus. Neither Shigella nor Salmonella genus were detected. In the majority of schools studied, it was found that food was of an adequate hygienic-sanitary standard. However, only municipal schools had the supervision of a technician responsible for school food. In the state schools, 60% had never been visited by a nutritionist and in these schools several procedures failed to comply with legal requirements. In most of the schools studied, the food served to students was within adequate standards, though the problems detected revealed the need for the implementation of Best Practices in the school environment.

Investigation into an unusual increase of human cases of Salmonella Goldcoast infection in Hungary in 2009.

We describe the outbreak investigation associated with an unusual increase in Salmonella Goldcoast cases in Hungary observed in autumn 2009, which included descriptive and analytical epidemiological studies and microbiological and veterinary investigations. Sixty cases were identified between 1 January 2009 and 1 March 2010, 50 of them from late July 2009 to January 2010. Of 50 S. Goldcoast isolates, 44 showed an indistinguishable pulsed-field gel electrophoresis profile. We conducted a matched case-control study that indicated a statistically significant association between S. Goldcoast infection and the consumption of pork cheese. The majority of cases (seven of nine) reporting consumption of this product belonged to a single family cluster. After removing six cases of this cluster, pork cheese still showed an elevated but non-significant risk for being a case in the univariable analysis (Mantel-Haenszel odds ratio (MH OR): 3.87, 95% confidence interval (CI): 0.38-39.47). A single S. Goldcoast isolate was identified during routine veterinary surveillance activities in 2009 in minced beef from a butcher's shop, originating from an abattoir where also pigs were slaughtered. We conclude that the outbreak was probably due to multiple sources of contaminated meat, probably pork, released on the market over a period of several months in 2009.

Assessment of hygiene standards and Hazard Analysis Critical Control Points implementation on passenger ships.

The level of hygiene on ferries can have impact on travellers' health. The aim of this study was to assess the hygiene standards of ferries in Greece and to investigate whether Hazard Analysis Critical Control Points (HACCP) implementation contributes to the hygiene status and particularly food safety aboard passenger ships. Hygiene inspections on 17 ferries in Greece were performed using a standardized inspection form, with a 135-point scale. Thirty-four water and 17 food samples were collected and analysed. About 65% (11/17) of ferries were scored with >100 points. Ferries with HACCP received higher scores during inspection compared to those without HACCP (p value <0.001). All 34 microbiological water test results were found negative and, from the 17 food samples, only one was found positive for Salmonella spp. Implementation of management systems including HACCP principles can help to raise the level of hygiene aboard passenger ships.

Microbiological assessment and evaluation of rehydration instructions on powdered infant formulas, follow-up formulas, and infant foods in Malaysia.

A total of 90 samples comprising powdered infant formulas (n=51), follow-up formulas (n=21), and infant foods (n=18) from 15 domestic and imported brands were purchased from various retailers in Klang Valley, Malaysia and evaluated in terms of microbiological quality and the similarity of rehydration instructions on the product label to guidelines set by the World Health Organization. Microbiological analysis included the determination of aerobic plate count (APC) and the presence of Enterobacteriaceae and Cronobacter spp. Isolates of interest were identified using ID 32E (bioMérieux France, Craponne, France). In this study, 87% of powdered infant formulas, follow-up formulas, and infant foods analyzed had an APC below the permitted level of <10(4) cfu/g. These acceptable APC ranged between <10(2) to 7.2×10(3) cfu/g. The most frequently isolated Enterobacteriaceae was Enterobacter cloacae, which was present in 3 infant formulas and 1 infant food tested. Other Enterobacteriaceae detected from powdered infant and follow-up formulas were Citrobacter spp., Klebsiella spp., and other Enterobacter spp. No Cronobacter species were found in any samples. Rehydration instructions from the product labels were collated and it was observed that none directed the use of water with a temperature >70°C for formula preparation, as specified by the 2008 revised World Health Organization guidelines. Six brands instructed the use of water at 40 to 55°C, a temperature range that would support the survival and even growth of Enterobacteriaceae.

Listeriosis outbreak caused by Latin-style fresh cheese, Bizkaia, Spain, August 2012.

Two cases of laboratory-confirmed listeriosis were detected in Bizkaia, Spain, at the end of August. The epidemiological investigation indicated that these two cases were associated with the consumption of Latin-style fresh cheese made from pasteurised milk in Portugal. Different batches of the same cheese were analysed and confirmed as contaminated with Listeria monocytogenes. The product was withdrawn from the market and the population was advised not to consume this kind of cheese.

Impact assessment of risk management interventions.

Much effort has been invested in the development and implementation of international recommendations to manage the risk of foodborne antimicrobial resistance, and monitoring programmes to measure bacterial antimicrobial resistance and antimicrobial product volumes. A variety of approaches have been recommended for various stakeholders in the food animal and food production sectors. Interestingly, much less consideration has been given to the establishment of success criteria for the individual interventions and even less for the cumulative effects, when all interventions are considered together as consecutive 'hurdles' along the food chain. The author explores the outcome and unforeseen consequences of these various interventions and appropriate methods that could provide data to assess their impact, as well as key learning experiences that should lead to refinements of such interventions in the future.

[The significance of some potentially pathogenic microorganisms in occurence of food toxicoinfections. Report 2. Assessment of the role of toxigenic strains of Bacillus cereus].

The data on nomenclature, classification and taxonomy of aerobic spore-forming Bacillus cereus are summarized. The main features of the two types of diseases, caused B. cereus, and statistical information on outbreaks of food-borne disease caused by B. cereus are presented. The detailed description of emetic toxin cereulide properties are given. The analysis of existing methods for detection of the presence of B. cereus and their toxins in foods are conducted. The data on the use of different cell models for studying the cytotoxic effects and the enterotoxigenic properties of B. cereus are described. Results of own researches allowed to conclude that certain types of products, primarily made from milk and vegetable raw materials, can be a source of transmission to humans of toxins produced by B. cereus. It is shown that in the absence of competing vegetative microflora increases the risk of accumulation of toxins produced by the most stable populations, including toxigenic spore B. cereus. Tested and proposed for the practical implementation of the dry culture media on the basis of the balanced growth and selective components, dyes and buffer mixtures. The developed environment were used for the isolation and identification B. cereus during microbial control these groups of foods.