Microbial quality of shrimp products of export trade produced from aquacultured shrimp

Bacteriological quality of individually quick frozen (IQF) shrimp products produced from aquacultured tiger shrimp (Penaeus monodon) has been analysed in terms of aerobic plate count (APC), coliforms, Escherichia coli, coagulase-positive staphylococci, Salmonella, and Listeria monocytogenes. Eight hundred forty-six samples of raw, peeled, and deveined tail-on (RPTO), 928 samples of cooked, peeled, and deveined tail-on (CPTO), 295 samples of headless, undeveined shell-on (HLSO), and 141 samples of raw, peeled, and deveined tail-off (RPND) shrimps were analysed for the above bacteriological parameters. Salmonella was isolated in only one sample of raw, peeled tail-on. Serotyping of the strain revealed that it was S. typhimurium. While none of the cooked, peeled tail-on shrimp samples exceeded the aerobic plate count (APC) of 105 colony forming units per gram (cfu/g), 2.5% of raw, peeled, tail-on, 6.4% of raw, peeled tail-off, and 7.5% of headless shell-on shrimp samples exceeded that level. Coliforms were detected in all the products, though at a low level. Prevalence of coliforms was higher in headless shell-on (26%) shrimps followed by raw, peeled, and deveined tail-off (19%), raw, peeled tail-on (10%), and cooked, peeled tail-on (3.8%) shrimps. While none of the cooked, peeled tail-on shrimp samples were positive for coagulase-positive staphylococci and E. coli, 0.6–1.3% of the raw, peeled tail-on were positive for staphylococci and E. coli, respectively. Prevalence of staphylococci was highest in raw, peeled tail-off (5%) shrimps and the highest prevalence of E. coli (4.8%) was noticed in headless shell-on shrimps. L. monocytogenes was not detected in any of the cooked, peeled tail-on shrimps. Overall results revealed that the plant under investigation had exerted good process control in order to maintain superior bacteriological quality of their products

Horizontal nutrient fluxes and production efficiencies in urban and peri-urban crop and livestock husbandry of Niamey, Niger

Urban and peri-urban agriculture (UPA) increasingly supplies food and non-food values to the rapidly growing West African cities. However, little is known about the resource use efficiencies in West African small-scale UPA crop and livestock production systems, and about the benefits that urban producers and retailers obtain from the cultivation and sale of UPA products. To contribute to filling this gap of knowledge, the studies comprising this doctoral thesis determined nutrient use efficiencies in representative urban crop and livestock production system in Niamey, Niger, and investigated potential health risks for consumers. Also assessed was the economic efficiency of urban farming activities. The field study, which was conducted during November 2005 to January 2008, quantified management-related horizontal nutrient flows in 10 vegetable gardens, 9 millet fields and 13 cattle and small ruminant production units. These farms, selected on the basis of a preceding study, represented the diversity of UPA crop and livestock production systems in Niamey. Based on the management intensity, the market orientation and especially the nutrient input to individual gardens and fields, these were categorized as high or low input systems. In the livestock study, high and low input cattle and small ruminant units were differentiated based on the amounts of total feed dry matter offered daily to the animals at the homestead. Additionally, economic returns to gardeners and market retailers cultivating and selling amaranth, lettuce, cabbage and tomato - four highly appreciated vegetables in Niamey were determined during a 6-months survey in forty gardens and five markets. For vegetable gardens and millet fields, significant differences in partial horizontal nutrient balances were determined for both management intensities. Per hectare, average annual partial balances for carbon (C), nitrogen (N), phosphorus (P) and potassium (K) amounted to 9936 kg C, 1133 kg N, 223 kg P and 312 kg K in high input vegetable gardens as opposed to 9580 kg C, 290 kg N, 125 kg P and 351 kg K in low input gardens. These surpluses were mainly explained by heavy use of mineral fertilizers and animal manure to which irrigation with nutrient rich wastewater added. In high input millet fields, annual surpluses of 259 kg C ha-1, 126 kg N ha-1, 20 kg P ha-1 and 0.4 kg K ha-1 were determined. Surpluses of 12 kg C ha-1, 17 kg N ha-1, and deficits of -3 kg P ha-1 and -3 kg K ha-1 were determined for low input millet fields. Here, carbon and nutrient inputs predominantly originated from livestock manure application through corralling of sheep, goats and cattle. In the livestock enterprises, N, P and K supplied by forages offered at the farm exceeded the animals’ requirements for maintenance and growth in high and low input sheep/goat as well as cattle units. The highest average growth rate determined in high input sheep/goat units was 104 g d-1 during the cool dry season, while a maximum average gain of 70 g d-1 was determined for low input sheep/goat units during the hot dry season. In low as well as in high input cattle units, animals lost weight during the hot dry season, and gained weight during the cool dry season. In all livestock units, conversion efficiencies for feeds offered at the homestead were rather poor, ranging from 13 to 42 kg dry matter (DM) per kg live weight gain (LWG) in cattle and from 16 to 43 kg DM kg-1 LWG in sheep/goats, pointing to a substantial waste of feeds and nutrients. The economic assessment of the production of four high value vegetables pointed to a low efficiency of N and P use in amaranth and lettuce production, causing low economic returns for these crops compared to tomato and cabbage to which inexpensive animal manure was applied. The net profit of market retailers depended on the type of vegetable marketed. In addition it depended on marketplace for amaranth and lettuce, and on season and marketplace for cabbage and tomato. Analysis of faecal pathogens in lettuce irrigated with river water and fertilized with animal manure indicated a substantial contamination by Salmonella spp. with 7.2 x 104 colony forming units (CFU) per 25 g of produce fresh matter, while counts of Escherichia coli averaged 3.9 x 104 CFU g-1. In lettuce irrigated with wastewater, Salmonella counts averaged 9.8 x 104 CFU 25 g-1 and E. coli counts were 0.6 x 104 CFU g-1; these values exceeded the tolerable contamination levels in vegetables of 10 CFU g-1 for E. coli and of 0 CFU 25 g-1 for Salmonella. Taken together, the results of this study indicate that Niamey’s UPA enterprises put environmental safety at risk since excess inputs of N, P and K to crop and livestock production units favour N volatilisation and groundwater pollution by nutrient leaching. However, more detailed studies are needed to corroborate these indications. Farmers’ revenues could be significantly increased if nutrient use efficiency in the different production (sub)systems was improved by better matching nutrient supply through fertilizers and feeds with the actual nutrient demands of plants and animals.

Heavy metal and microbial loads in sewage irrigated vegetables of Kabul, Afghanistan

Little is known about the heavy metal and microbial contamination of vegetables produced in Central Asian cities. We therefore measured the concentration of cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn) and of faecal pathogens (Coliform bacteria, Salmonella sp., Shigella sp., Ascaris lubricoides, Entamoeba sp. and pinworms [Oxyuris vermicularis syn. Enterobius vermicularis]) in soil, irrigation water, and marketed vegetables of Kabul City, Afghanistan. Leaf Pb and Zn concentrations of leafy vegetables were with 1–5 and 33–160 mg kg^{-1} dry weight (DW) several-fold above respective international thresholds of 0.3 mg Pb kg^{-1} and 50 mg Zn kg^{-1}. The tissue concentration of Cu was below threshold limits in all samples except for spinach in one farm. Above-threshold loads of microbes and parasites on vegetables were found in five out of six gardens with coliforms ranging from 0.5–2 × 10^7 cells 100g^{-1} fresh weight (FW), but no Salmonella and Shigella were found. Contamination with 0.2 × 10^7 eggs 100g^{-1} FW of Ascaris was detected on produce of three farms and critical concentrations of Entamoeba in a single case, while Oxyuris vermicularis, and Enterobius vermicularis were found on produce of three and four farms, respectively. Irrigation water had Ascaris, Coliforms, Salmonella, Shigella, Entamoeba, and Oxyuris vermicularis syn. Enterobius vermicularis ranging from 0.35 × 10^7 to 2 × 10^7 cells l^{-1}. The heavy metal and microbial loads on fresh UPA vegetables are likely the result of contamination from rising traffic, residues of the past decades of war and lacking treatment of sewage which needs urgent attention.

Laboratorio virtual para la enseñanza y aprendizaje de la identificación de microorganismos.

El trabajo, de momento, no ha sido publicado

Caracterització de Lactobacils d'origen intestinal i avaluació in vivo del seu poder probiòtic en pollastres

Una soca de Lactobacillus salivarius resistent a la rifampicina, CTC2197, es va assajar com a probiòtic en pollastres, estudiant la seva capacitat de prevenir la colonització de Salmonella enteritidis C-114 en pollastres. Quan la soca probiòtica es va administrar via oral juntament amb S.enteritidis C-114 directament al proventricle en pollets Leghorn de 1 dia, el patògen fou eliminat completament després de 21 dies. Els mateixos resultats es van obtenir quan la soca es va administrar a través del menjar i l'aigua a més de la inoculació directa al proventricle. La inclusió de L.salivarius CTC2197 en el menjar del primer dia va mostrar que una concentració de 105 UFC g-1 era suficient per assegurar la colonització dels tracte gastrointestinal dels pollets després de 1 setmana. No obstant, entre els 21 i 28 dies, L.salivarius CTC2197 no va ser detectable en el tracte gastrointestinal d'alguns pollets, mostrant que seria necessària més d'una dosis per assegurar la seva presència fins al final de l'etapa d'engreix. La liofilització i la congelació per glicerol o llet descremada com a agents crioprotector, van semblar mètodes adequats per preservar la soca probiòtica. La inclusió de L.salivarius CTC2197 en un pinso comercial va semblar ser un bon mètode per subministrar-lo en granja, tot i que la soca va mostrar sensibilitat a les temperatures utilitzades durant l'emmagatzematge del pinso i a les incubadores dels pollets. A més, la supervivència va millorar després de diverses reinoculacions en pinso.

Development of molecular-based techniques for the detection, identification and quantification of food-borne pathogens

La presencia de microorganismos patógenos en alimentos es uno de los problemas esenciales en salud pública, y las enfermedades producidas por los mismos es una de las causas más importantes de enfermedad. Por tanto, la aplicación de controles microbiológicos dentro de los programas de aseguramiento de la calidad es una premisa para minimizar el riesgo de infección de los consumidores. Los métodos microbiológicos clásicos requieren, en general, el uso de pre-enriquecimientos no-selectivos, enriquecimientos selectivos, aislamiento en medios selectivos y la confirmación posterior usando pruebas basadas en la morfología, bioquímica y serología propias de cada uno de los microorganismos objeto de estudio. Por lo tanto, estos métodos son laboriosos, requieren un largo proceso para obtener resultados definitivos y, además, no siempre pueden realizarse. Para solucionar estos inconvenientes se han desarrollado diversas metodologías alternativas para la detección identificación y cuantificación de microorganismos patógenos de origen alimentario, entre las que destacan los métodos inmunológicos y moleculares. En esta última categoría, la técnica basada en la reacción en cadena de la polimerasa (PCR) se ha convertido en la técnica diagnóstica más popular en microbiología, y recientemente, la introducción de una mejora de ésta, la PCR a tiempo real, ha producido una segunda revolución en la metodología diagnóstica molecular, como pude observarse por el número creciente de publicaciones científicas y la aparición continua de nuevos kits comerciales. La PCR a tiempo real es una técnica altamente sensible -detección de hasta una molécula- que permite la cuantificación exacta de secuencias de ADN específicas de microorganismos patógenos de origen alimentario. Además, otras ventajas que favorecen su implantación potencial en laboratorios de análisis de alimentos son su rapidez, sencillez y el formato en tubo cerrado que puede evitar contaminaciones post-PCR y favorece la automatización y un alto rendimiento. En este trabajo se han desarrollado técnicas moleculares (PCR y NASBA) sensibles y fiables para la detección, identificación y cuantificación de bacterias patogénicas de origen alimentario (Listeria spp., Mycobacterium avium subsp. paratuberculosis y Salmonella spp.). En concreto, se han diseñado y optimizado métodos basados en la técnica de PCR a tiempo real para cada uno de estos agentes: L. monocytogenes, L. innocua, Listeria spp. M. avium subsp. paratuberculosis, y también se ha optimizado y evaluado en diferentes centros un método previamente desarrollado para Salmonella spp. Además, se ha diseñado y optimizado un método basado en la técnica NASBA para la detección específica de M. avium subsp. paratuberculosis. También se evaluó la aplicación potencial de la técnica NASBA para la detección específica de formas viables de este microorganismo. Todos los métodos presentaron una especificidad del 100 % con una sensibilidad adecuada para su aplicación potencial a muestras reales de alimentos. Además, se han desarrollado y evaluado procedimientos de preparación de las muestras en productos cárnicos, productos pesqueros, leche y agua. De esta manera se han desarrollado métodos basados en la PCR a tiempo real totalmente específicos y altamente sensibles para la determinación cuantitativa de L. monocytogenes en productos cárnicos y en salmón y productos derivados como el salmón ahumado y de M. avium subsp. paratuberculosis en muestras de agua y leche. Además este último método ha sido también aplicado para evaluar la presencia de este microorganismo en el intestino de pacientes con la enfermedad de Crohn's, a partir de biopsias obtenidas de colonoscopia de voluntarios afectados. En conclusión, este estudio presenta ensayos moleculares selectivos y sensibles para la detección de patógenos en alimentos (Listeria spp., Mycobacterium avium subsp. paratuberculosis) y para una rápida e inambigua identificación de Salmonella spp. La exactitud relativa de los ensayos ha sido excelente, si se comparan con los métodos microbiológicos de referencia y pueden serusados para la cuantificación de tanto ADN genómico como de suspensiones celulares. Por otro lado, la combinación con tratamientos de preamplificación ha resultado ser de gran eficiencia para el análisis de las bacterias objeto de estudio. Por tanto, pueden constituir una estrategia útil para la detección rápida y sensible de patógenos en alimentos y deberían ser una herramienta adicional al rango de herramientas diagnósticas disponibles para el estudio de patógenos de origen alimentario.

Lactic acid bacteria as bioprotective agents against foodborne pathogens and spoilage microorganisms in fresh fruits and vegetables

La present tesi doctoral es centra en l'aplicació dels bacteris de l'àcid lactic (BAL) com a agents bioprotectors davant microorganismes patògens i deteriorants.Es van aïllar i seleccionar BAL de fruites i hortalisses fresques i es van assajar in vitro davant 5 microorganismes fitopatògens i 5 patògens humans.Es van realitzar assajos d'eficàcia en pomes Golden Delicious amb tots els aïllats enfront les infeccions causades pel fong Penicillium expansum. La soca més eficaç era Weissella cibaria TM128, que reduïa el diàmetre de les infeccions en un 50%.Les soques seleccionades es van assajar enfront els patògens Salmonella typhimurium, Escherichia coli i Listeria monocytogenes en enciams Iceberg i pomes Golden Delicious.Els BAL interferien eficientment amb el creixemet de S. typhimurium, and L. monocytogenes, però van mostrar poc efecte enfront E. coli.Finalment, es van realitzar assajos dosi-resposta amb les soques Leuconostoc mesenteroides CM135, CM160 and PM249 enfront L. monocytogenes. De totes les soques assajades, la soca CM160 va ser la més efectiva.

Importância da utilização das ferramentas de gestão da qualidade para a produção de alimentos seguros : análise de uma unidade de alimentação e nutrição (UAN) na cidade de Belém - PA

As empresas do segmento de alimentação coletiva cresceram significativamente nos últimos anos, influenciadas, principalmente, pela exigência dos usuários que têm como referência a grande variedade de produtos e serviços disponíveis no consumo de refeições fora do âmbito doméstico. A quantidade de pessoas que se alimenta fora de casa é cada vez maior, devido a fatos como crescimento das cidades, inserção da mulher no mercado de trabalho e tempo indisponível para o preparo das refeições. Para garantir a segurança dos alimentos oferecidos, é necessária a aplicação de ferramentas da qualidade como as Boas Práticas, procedimentos operacionais padronizados e outras, que devem ser adotadas na linha de processamento de alimentos com o objetivo de evitar possíveis doenças transmitidas por alimentos. O objetivo da presente pesquisa foi de analisar a importância da utilização das ferramentas da qualidade no processamento das preparações culinárias produzidas em uma Unidade de Alimentação e Nutrição (UAN) na cidade de Belém do Pará. Foram coletadas 50 amostras de preparações culinárias produzidas no estabelecimento onde foi observada contaminação de 36% por coliforme a 45ºC,16% por S. aureus e 4% por salmonella spp., foi aplicado um check-list (lista de verificação) baseada na legislação vigente, de forma a identificar as não-conformidades apresentadas pelos estabelecimentos.Os resultados mostraram que 47,4% dos itens de boas práticas não estão sendo aplicadas no estabelecimento. Observou-se durante a pesquisa na Unidade de Alimentação e Nutrição que as ferramentas da qualidade não são aplicadas e algumas são aplicadas inadequadamente durante o processamento das preparações culinárias, concorrendo para a possibilidade de ofertar aos comensais de produtos impróprios para o consumo. Conclui-se que a utilização das ferramentas da qualidade são importantes recursos para a obtenção da garantia da qualidade que podem ser aprimoradas através do desenvolvimento de treinamentos de habilidades e percepção para trabalhar atitudes de cada colaborador dentro da linha de produção de alimentos seguros.

Optimal protection of stabilised dry live bacteria from bile toxicity in oral dosage forms by bile acid adsorbent resins

We previously found that dried live bacteria of a vaccine strain can be temporarily sensitive to bile acids and suggested that Bile Adsorbing Resins (BAR) can be used in oral vaccine tablets to protect dried bacteria from intestinal bile. Here, we report a quantitative analysis of the ability of BAR to exclude the dye bromophenol blue from penetrating into matrix tablets and also sections of hard capsule shells. Based on this quantitative analysis, we made a fully optimised formulation, comprising 25% w/w of cholestyramine in Vcaps™ HPMC capsules. This gave effectively 100% protection of viability from 4% bile, with 4200-fold more live bacteria recovered from this formulation compared to unprotected dry bacteria. From the image analysis, we found that the filler material or compaction force used had no measurable effect on dye exclusion but did affect the rate of tablet hydration. Increasing the mass fraction of BAR gave more exclusion of dye up to 25% w/w, after which a plateau was reached and no further dye exclusion was seen. More effective dye exclusion was seen with smaller particle sizes (i.e. cholestyramine) and when the BAR was thoroughly dried and disaggregated. Similar results were found when imaging dye penetration into capsule sections or tablets. The predictions of the dye penetration study were tested using capsules filled with dried attenuated Salmonella vaccine plus different BAR types, and the expected protection from bile was found, validating the imaging study. Surprisingly, depending on the capsule shell material, some protection was given by the capsule alone without adding BAR, with Vcaps™ HPMC capsules providing up to 174-fold protection against 1% bile; faster releasing Vcaps Plus™ HPMC capsules and Coni Snap™ gelatin capsules gave less protection.

Formulation of a live bacterial vaccine for stable room temperature storage results in loss of acid, bile and bile salt resistance

Live bacterial vaccines have great promise both as vaccines against enteric pathogens and as heterologous antigen vectors against diverse diseases. Ideally, room temperature stable dry formulations of live bacterial vaccines will allow oral vaccination without cold-chain storage or injections. Attenuated Salmonella can cross the intestinal wall and deliver replicating antigen plus innate immune activation signals directly to the intestinal immune tissues, however the ingested bacteria must survive firstly gastric acid and secondly the antimicrobial defences of the small intestine. We found that the way in which cells are grown prior to formulation markedly affects sensitivity to acid and bile. Using a previously published stable storage formulation that maintained over 10% viability after 56 days storage at room temperature, we found dried samples of an attenuated S. typhimurium vaccine lost acid and bile resistance compared to the same bacteria taken from fresh culture. The stable formulation utilised osmotic preconditioning in defined medium plus elevated salt concentration to induce intracellular trehalose accumulation before drying. Dried bacteria grown in rich media without osmotic preconditioning showed more resistance to bile, but less stability during storage, suggesting a trade-off between bile resistance and stability. Further optimization is needed to produce the ultimate room-temperature stable oral live bacterial vaccine formulation.

Modelling risk perception and trust in food safety information within the theory of planned behaviour

This study suggests a statistical strategy for explaining how food purchasing intentions are influenced by different levels of risk perception and trust in food safety information. The modelling process is based on Ajzen's Theory of Planned Behaviour and includes trust and risk perception as additional explanatory factors. Interaction and endogeneity across these determinants is explored through a system of simultaneous equations, while the SPARTA equation is estimated through an ordered probit model. Furthermore, parameters are allowed to vary as a function of socio-demographic variables. The application explores chicken purchasing intentions both in a standard situation and conditional to an hypothetical salmonella scare. Data were collected through a nationally representative UK wide survey of 533 UK respondents in face-to-face, in-home interviews. Empirical findings show that interactions exist among the determinants of planned behaviour and socio-demographic variables improve the model's performance. Attitudes emerge as the key determinant of intention to purchase chicken, while trust in food safety information provided by media reduces the likelihood to purchase. (C) 2006 Elsevier Ltd. All rights reserved.

Consumption of welfare-friendly food products in Great Britain, Italy and Sweden, and how it may be influenced by consumer attitudes to, and behaviour towards, animal welfare attributes

The interest in animal welfare and welfare-friendly food products has been increasing in Europe over the last 10 years. The media, highlighting traditional farming methods and food scares such as those related to salmonella, bovine spongiform encephalopathy/variant Creutzfeldt-Jakob disease (BSE) and avian influenza, have brought the methods of animal farming to public attention. Concerns about farm animal welfare are reflected in the increase in the number of vegetarians and vegans and an increase in consumers wishing to purchase food which is more animal welfare-friendly. This paper considers consumers’ attitudes to animal welfare and to marketing practices, such as product labelling, welfare grading systems and food assurance marks using comparative data collected in a survey of around 1500 consumers in each of Great Britain, Italy and Sweden as part of the EU-funded Welfare Quality research project. The findings suggest a need for the provision of improved consumer information on the welfare provenance of food using appropriate product labelling and other methods.

Food scares and trust: A European study

The complex interactions between the determinants of food purchase under risk are explored using the SPARTA model, based on the theory of planned behaviour, and estimated through a combination of multivariate statistical techniques. The application investigates chicken consumption choices in two scenarios: ( a) a 'standard' purchasing situation; and (b) following a hypothetical Salmonella scare. The data are from a nationally representative survey of 2,725 respondents from five European countries: France, Germany, Italy, the Netherlands and the United Kingdom. Results show that the effects and interactions of behavioural determinants vary significantly within Europe. Only in the case of a food scare do risk perceptions and trust come into play. The policy priority should be on building and maintaining trust in food and health authorities and research institutions, while food chain actors could mitigate the consequences of a food scare through public trust. No relationship is found between socio-demographic variables and consumer trust in food safety information.

Purification and characterization of the flagellar basal body of Rhodobacter sphaeroides

Flagellar hook-basal body (HBB) complexes were purified from Rhodobacter sphaeroides. The HBB was more acid labile but more heat stable than that of Salmonella species, and protein identification revealed that HBB components were expressed only from one of the two sets of flagellar gene clusters on the R. sphaeroides genome, under the heterotrophic growth conditions tested here.