Este trabalho teve como objetivo determinar as respostas fisiológicas, físicoquímicas e microbiológicas associadas ao processamento mínimo do rabanete, sendo que para isso foram realizados 5 experimentos. No primeiro experimento, foram avaliadas taxa respiratória e produção de etileno de raízes em retalhos, mantidas a 5ºC (±1ºC) e 90% (±5%) UR por 10 dias. No 10º dia, os rabanetes em retalhos apresentaram uma taxa de respiração 149% superior à verificada nas raízes inteiras (70,35 e 28,23 mL CO2 kg-1 h-1, respectivamente). Não foi detectada produção de etileno. No segundo, foram avaliados os aspectos microbiológicos após o corte e no 10º dia, de dois tratamentos (uma e duas sanitizações), em rabanete minimamente processado mantido a 5ºC (±1ºC) e 90% (±5%) UR. As contagens de bactérias psicrotróficas no tratamento com duas sanitizações mantiveram-se abaixo do limite aceitável; no tratamento com uma sanitização obteve-se, no 10º dia de armazenamento, 5,8 x 106 UFC/g, limite máximo recomendado. Não foi detectada presença de coliformes totais e Salmonella. No terceiro experimento, estudaram-se dois tipos de corte (rodelas e retalhos) e três temperaturas de armazenamento (1, 5 e 10ºC), por 10 dias, analisando-se taxa respiratória, produção de etileno e parâmetros físico-químicos. Após 12 horas do processamento obteve-se, nas raízes em retalhos, 0,14, 0,38 e 0,70 µL C2H4 kg-1 h-1 a 1, 5 e 10ºC, respectivamente. No 10º dia, as raízes inteiras a 1ºC apresentaram a menor taxa respiratória (5,72 mL CO2 kg- 1 h-1) e as raízes em retalhos a 10ºC, a maior taxa (26,71 mL CO2 kg-1h-1). Essas, também, apresentaram um decréscimo no teor de vitamina C. No quarto experimento, avaliaram-se embalagens de filme de PVC (14 e 17 µm de espessura) envolvendo bandejas de poliestireno expandido; filmes plásticos de polietileno de baixa densidade (PEBD) com 20 µm; e embalagens de tereftalato de polietileno (PET), mantidas a 5ºC (±1ºC) e 90% (±5%) UR por 10 dias, sendo realizadas análises de O2, CO2 e físicoquímicas. O teor de equilíbrio de O2 nas embalagens de PVC foi de 12 e 11% para as espessuras 14 e 17 µm, respectivamente. O PEBD apresentou concentrações muito baixas de O2 (0,08% no 8º dia), tendo como conseqüência processos fermentativos. Entre o 2º e 10º dia, o nível de CO2 no interior da embalagem de PVC com 14 µm variou de 3,43 a 2,43%; na PET de 1,78 a 4% e no PEBD de 12 a 15,2%. Os valores de luminosidade (L*), decresceram no decorrer do experimento, indicando escurecimento do produto. No quinto experimento foram estudados antioxidantes (ácido cítrico e ascórbico) em rabanete minimamente processado mantido a 5ºC (±1ºC) e 90% (±5%) UR por 10 dias, avaliando-se taxa respiratória e análises físico-químicas. O tratamento com ácido ascórbico apresentou a maior taxa respiratória nas 4 primeiras horas após o processamento. No 10º dia, obteve-se 34,18; 30,54; 21,31 e 2,22 mL CO2 kg-1 h-1 nos tratamentos com ácido ascórbico, ácido cítrico + ácido ascórbico, controle e ácido cítrico, respectivamente. Os valores de L*, de maneira geral, foram decrescendo ao longo do armazenamento. Os valores de a* mostraram que os tratamentos com ácido cítrico conferiram uma coloração fortemente avermelhada às raízes. Nenhum dos tratamentos evitou o escurecimento do rabanete minimamente processado. / This research was carried out with the objective to determine the physicochemical, physiological and microbiological alterations associated to the fresh cutting of radish. Five experiments were evaluated. In the first, respiratory rate and ethylene production were evaluated in radishes shredded, and stored at 5ºC (±1ºC) and 90% (±5%) RH during 10 days. In the 10th day, the fresh cut radishes showed respiratory rate 149% higher if compared to the whole radishes (70.35 and 28.23 mL CO2 kg-1 h-1, respectively). Ethylene production was not detected. In the second experiment, microbiological evaluations after the cut and in the 10th day, were carried out in fresh cut radishes submitted to one or two sanitation and stored at 5ºC (±1ºC) and 90% (±5%) RH. The numbers of psychotropic bacteria in the treatment with two sanitations stayed on the acceptable limits. In the treatment with one sanitation it was obtained 5.8 x 106 CFU/g in the 10th of storage that is the maximum limit recommended. It was not detected the presence of total coliform and Salmonella. In the third experiment, it was studied two types of cut (slices and shredded) and three storage temperatures (1, 5 and 10ºC). Respiratory rate, ethylene production and physicochemical parameters were evaluated during 10 days. After 12 hours of the processing, it was obtained 0.14, 0.38 and 0.70 mL CO2 kg-1 h-1 to 1, 5 and 10ºC, respectively, in the shredded radishes. In the 10th day, the whole radishes at 1ºC showed the lower smallest respiratory rate (5.72 mL CO2 kg-1 h-1), while whole radishes showed the higher rate at 10ºC (26.71 mL CO2 kg-1 h-1). Also, in the last treatment it was verified a decrease in the contents of vitamin C. In the fourth experiment were evaluated different types of packages in fresh cut radishes stored at 5ºC and 90% RH during 10 days. The packages used were: PVC films with 14 and 17 µm thickness overwrapping expanded polystyrene trays, low density polyethylene film (LDPE) with 20 µm thickness, and polyethylene terephthalate (PET). Concentrations of O2 and CO2 within package and physicochemical analysis were evaluated. The equilibrium concentration of O2 in the package of PVC was 12 and 11% for the 14 and 17 µm thickness, respectively. The LDPE presented very low concentrations of O2 (0.08% on the 8th day), having as consequence fermentative processes. From 2nd to 10th day, the level of CO2 inside the packing of PVC film with 14 µm changed from 3.43 to 2.43%, in PET from 1.78 to 4% and in LDPE from 12 to 15.2%. The values of lightness (L*) decreased in all treatments during storage. In the fifth experiment, it were studied the use of antioxidants (citric and ascorbic acid) in fresh cut radishes, stored at 5ºC and 90% RH during 10 days. Respiratory rate and physicochemical analysis were evaluated. The treatment with ascorbic acid presented the higher respiratory rate in the first 4 hours after the processing. In the 10th day, it was obtained 34.18; 30.54; 21.31 and 2.22 mL CO2 kg-1h-1 in the treatments with ascorbic acid, citric acid + ascorbic acid, control and citric acid, respectively. The treatments with ascorbic acid showed an increment in TSS content on the 2nd day of storage. The values of L* decreased during cold storage. The values of the a* showed that treatments with citric acid presented a strongly red coloration in the roots minimally processed. None of the treatments have avoided the browning of the shredded fresh cut radishes cold stored.

Made available in DSpace on 2014-07-29T15:26:21Z (GMT). No. of bitstreams: 1 Liana Jayme borges.pdf: 1844496 bytes, checksum: 818c982e89c90b277c44959cde90066a (MD5) Previous issue date: 2010-03-19 / Enteral feeding means the nutrition for special purposes, with controlled intake of nutrients. The advantages of its use often become secondary to complications arising from its contamination, which may be associated with infectious complications. The microbial contamination of enteral feeding may occur during all steps being the handling, particularly critical. Considering the importance of enteral feeding as a therapeutic tool in hospitals and the need to guarantee the microbiological quality of the products offered to critical patients, the present work aimed to evaluate the hygienic and sanitary quality of diets and their ingredients and to identify and characterize phenotypic and genotypically, using the antibiogram and pulsed-field gel electrophoresis, strains of Escherichia coli and Staphylococcus aureus obtained from handlers hands and noses, water, module and enteral nutrition from two public hospital in Goiânia, Brazil in order to investigate the probable source of microbological contamination. A total of 80 samples were collected from enteral nutrition and 140 from hands and noses of handlers involved in the diets manufacturing in hospital 1 (H1), between october/2007 and november/2008 and 80 samples from enteral nutrition and 80 from hands and noses of handlers in hospital 2 (H2), between october/2008 and november/2008. From both hospitals were collected 40 samples from water and module. The samples were submitted to microbiological analysis to verify the presence and numbers of pathogenic and indicator microorganisms. E. coli and S. aureus strains were submitted to antibiogram and PFGE. According to antibiogram, all S.aureus isolates (15) from H1 were susceptible to oxacillin, vancomycin, ciprofloxacin and gentamicin. Resistence profile was observed in 10 (66.7%) isolates for penicillin, four (26.7%) isolates for tetracycline and nine (60.0%) isolates for erythromycin, allowing to classify the strains in six different phenotypes (A-F), but it was not efficient for the determination of the bacterial source for the diets. In the H1, all (08) E. coli strains were susceptible to trimethoprim, ciprofloxacin, cephalothin, gentamicin, ceftazidime and tetracycline. Resistence was observed in six (75.0%) isolates for ampicilin. In H2, all strains isolated (12) were susceptible to trimethoprim, ciprofloxacin, gentamicin and ceftazidime and resistence was observed in 11 isolates (91.7%) for cephalothin and 12 (100.0%) for tetracycline and ampicillin, grouping them into five different phenotypes (A-D). Microorganisms showed the same phenotypic profile from handlers and diet samples (phenotypes A and C), suggesting that in these cases, the source of microorganisms for the final product was the food handler. The genotypic typing of S. aureus strains by PFGE generated seven different DNA banding profiles and the E. coli genotyping generated five profiles. Based on the results, two E. coli strains isolated from diets were identical to one strain isolated from food handler from H2 and two of S. aureus isolated from diets were identical to one strain isolated from food handler from H1. This study shows that the enteral feedings showed unsatisfactory sanitary-hygienic conditions in both hospitals and the hand contact is probably one of the sources of greatest significance for enteral diets contamination in the hospital environment. / Entende-se por nutrição enteral a alimentação para fins especiais, com ingestão controlada de nutrientes. As vantagens oferecidas pelo seu emprego muitas vezes tornam-se secundárias às complicações derivadas de sua utilização como a contaminação, que pode estar associada a complicações infecciosas. A contaminação microbiana das fórmulas enterais pode ocorrer em diversas etapas, sendo a manipulação uma etapa especialmente crítica. Tendo em vista a importância da dieta enteral como medida terapêutica em hospitais e a necessidade de se ofertar produtos com qualidade assegurada, devido aos prejuízos que a mesma pode causar aos pacientes, caso esteja contaminada, o objetivo deste estudo foi avaliar a qualidade higiênico-sanitária das dietas e seus ingredientes e caracterizar fenotipicamente, utilizando o antibiograma e, genotipicamente, através da eletroforese em gel em campo pulsado (pulsed-field gel electrophoresis (PFGE), isolados de Escherichia coli e Staphylococcus aureus a partir de manipuladores, água, módulo em pó e dieta enteral de dois hospitais públicos de Goiânia-GO visando estabelecer a possível fonte de microrganismos para o produto final. Um total de 80 amostras de dieta enteral e 140 swabs de mãos e fossas nasais de manipuladores foram coletadas no hospital 1 (H1) entre outubro/2007 e novembro/2008 e 80 amostras de dieta enteral e 80 swabs de mãos e fossas nasais no hospital 2 (H2) entre novembro/2008 e dezembro/2008. Nos dois hospitais foram coletadas também 40 amostras de água e módulo. Foram realizadas análises microbiológicas para contagem de microrganismos indicadores e potencialmente patogênicos. Os isolados de E. coli e S.aureus foram submetidos ao antibiograma e PFGE. De acordo com o antibiograma, todas as cepas de S. aureus isoladas (15) no H1 foram sensíveis à oxacilina, vancomicina, ciprofloxacina e gentamicina. O padrão de resistência foi observado em 10 (66,7%) isolados para penicilina, quatro (26,7%) para tetraciclina e nove (60,0%) para eritromicina, agrupando-os em seis diferentes perfis fenotípicos (A F). Porém, a técnica não foi eficiente em determinar a origem da contaminação das dietas. Para as 20 cepas isoladas de E. coli do H1 e H2, todas (8) do H1 foram sensíveis ao trimetoprim, ciprofloxacina, cefalotina, gentamicina, ceftazidima e tetraciclina. Resistência foi observada em seis (75,0%) isolados para a ampicilina. No H2 todas as cepas isoladas (12) foram sensíveis ao trimetoprim, ciprofloxacina, gentamicina e ceftazidima e resistência foi observado em 11 isolados (91,7%) para a cefalotina e 12 (100,0%) para a tetraciclina e ampicilina, sendo agrupadas em quatro diferentes perfis fenotípicos (A D). Os fenótipos A e C apresentaram microrganismos com o mesmo perfil fenotípico provenientes de manipuladores e dieta, sugerindo que nestes casos, a fonte de microrganismos para o produto final seria os manipuladores. A tipificação genotípica por PFGE originou sete perfis eletroforéticos diferentes para as cepas de S.aureus e cinco para as cepas de E. coli. De acordo com os resultados, duas cepas de E. coli isoladas da dieta foram idênticas a uma cepa isolada do manipulador do H2 e duas cepas de S.aureus isoladas da dieta foram iguais a uma cepa do manipulador do H1. Os dados obtidos neste estudo permitem concluir que as dietas enterais apresentaram condições higiênico-sanitárias insatisfatórias em ambos os hospitais e que o manipulador é provavelmente, uma das fontes de maior significância para a contaminação da dieta enteral em ambiente hospitalar.

Salmonella é um dos mais importantes patógenos causadores de enfermidades transmitidas por alimentos (ETA) no Brasil e em outros países. Devido ao surgimento de fenótipos multi-resistentes (MOR) a agentes antimicrobianos em Salmonella, a caracterização dos genes envolvidos neste processo, sua localização e diversidade são importantes para identificação e compreensão dos fatores envolvidos na resistência. O objetivo deste trabalho foi caracterizar determinantes genéticos de virulência, resistência e integrons classe 1 presentes em cepas de diferentes sorotipos de Salmonella enterica multi-resistentes a antibióticos, isoladas a partir de alimentos de origem suína, avícola e fontes relacionadas. As cepas empregadas pertenceram a nove perfis PFGE distintos com a enzima Xbal, com similaridade genética variando de 38% a 68%. Integrons classe 1 foram detectados em 9 (45%) das 20 cepas de Salmonella enterica, incluindo cinco diferentes sorotipos: Brandenburg, Panama, Agona, Mbandaka e Alachua, e variando de 0,7Kb a 2,7Kb. Os genes de resistência aadA, sul1, sul2, tetA, dhfr, qacEΔl e blatem, que conferem resistência a aminoglicosídeos, sulfonamidas, tetraciclinas, compostos de amônio quaternário e β-lactâmicos, respectivamente, foram identificados no interior dos integrons, no cromossomo bacteriano, ou em ambos. Os genes aadB, floR, tetB e tetG não foram detectados. A resistência às quinolonas foi caracterizada nas 11 cepas que apresentaram resistência ao ácido nalidixico pela análise do gene gyrAM e mutações Ser-83-Fen foram confirmadas após sequenciamento das amostras. Estudos de conjugação demonstraram que apenas uma cepa de Salmonella Mbandaka foi capaz de transferir o gene sul2, para uma cepa de E. coli K12. Com relação ao perfil de virulência, as 20 cepas de Salmonella enterica foram caracterizadas e os genes slyA, invA, sopB e aceK estiveram presentes em 100% delas e o gene h-1i esteve presente em 18 cepas (90%). O gene spvC não foi detectado nas cinco cepas que possuíam plasmídeos. Os dados do presente estudo sugerem que alimentos de origem animal podem ser considerados como reservatórios de cepas de Salmonella enterica virulentas, resistentes a antimicrobianos e apresentando integrons classe 1. Isto caracteriza os produtos de origem suína e avícola como uma importante fonte de patógenos multi-resistentes para humanos. / Salmonella is one of the most important foodborne pathogens in Brazil and worldwide. Due to the emerging of multiresistant phenotypes in Salmonella the characterization of the genes involved in this process, their localization and diversity are important for identifying and understanding the factors involved in the resistance. The purpose of this study was to characterize virulence and antimicrobial determinants in different serovars of antibiotic multiresistant Salmonella enterica strains isolated from pork, poultry and related sources. The isolates belonged to nine different PFGE profiles obtained with Xbal restriction enzyme and showing genetic similarity ranging from 38% to 68%. Class 1 integrons were detected in 9 (45%) of 20 S. enterica strains ranging in size from 0,7Kb to 2,7Kb and comprising five different serotypes: Brandenburg, Panama, Agona, Mbandaka and Alachua. Resistance genes aadA, qacEΔl, sul1, tetA, sul2, dhfr, blatem, that confer resistance to aminoglicosides, sulphonamides, tetracyclines, ammonium quaternary compounds and beta-Iactams, respectively, were identified within class 1 integrons, chromosome, or both. Genes aad8, floR, tetB and tetG were not detected. The resistance to quinolones was characterized in 11 strains that showed resistance to nalidixic acid analyzing gyrA genes and Ser-83-Fen mutations were confirmed after sequencing of the samples. Conjugation studies demonstrated that only one S. Mbandaka strain was able to transfer sul2 gene to the E.coli K12. Regarding virulence profile Salmonella enterica strains were characterized and PCR analysis revealed the presence of the virulence genes invA, aceK, sop8, slyA in all isolates and the presence of virulence gene h-1i in 18 (90%) of them. The spvC gene was not detected in the five strains that harbored plasmids. The data of the present study suggest that foods of animal origin can be considered reservoirs of Salmonella enterica that are virulent, resistant and show class 1 integrons. This characterizes pork and poultry products as important sources of multi-resistant pathogens to human beings.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-08T12:54:40Z No. of bitstreams: 2 Dissertação - Natália Menezes Silva - 2016.pdf: 5000977 bytes, checksum: e00d87ceb9bd95951496870e4475fc1b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-08T12:56:13Z (GMT) No. of bitstreams: 2 Dissertação - Natália Menezes Silva - 2016.pdf: 5000977 bytes, checksum: e00d87ceb9bd95951496870e4475fc1b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-08T12:56:13Z (GMT). No. of bitstreams: 2 Dissertação - Natália Menezes Silva - 2016.pdf: 5000977 bytes, checksum: e00d87ceb9bd95951496870e4475fc1b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-05-02 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The pulp processing is a possibility for use of fruits by traditional people like the quilombolas, inserted in a context in which native fruits are little consumed. Therefore, the hygienic and sanitary quality is essential in the production of safe food. This study aimed to evaluate the processing and hygienic and sanitary conditions of fruits and fruit pulps in quilombola community. The survey was conducted in the Community of the Remnant of Quilombo do Pombal, Goiás, Brazil. To evaluate the physical and functional conditions was applied a checklist before and after the implementation of structural adjustments, training and implementation of the Manual of Good Practices and Standard Operating Procedures. Data were categorized according to the percentage of adequacy. Microbiological analyzes of 32 samples were taken including, fruits of Cerrado found in the community and fruits and their pulps in different steps of the flowchart production. The hazards and critical control points were listed in the production flow chart. The physical and functional diagnostic performed indicated average compliance of blocks of 30.18%, being classified in Group 3, unsatisfactory. After interventions, there was 62.13% of compliance, with classification in Group 2, regular (p <0.05). The intervention effect was significant (p <0.05) for five of the seven blocks evaluated. Microbiological analysis indicated that all the native fruits collected are in accordance with the recommendations of the current health legislation. Fruits and pulps included into processing flowchart presented in accordance with sanitary standards for fecal coliform count and detection of Salmonella spp, however, the mold count and yeast in eight pulps analyzed indicated that five of them had counts above the established by legislation. The results suggest fruit contamination since the harvest and multiplication of such micro-organisms in the remaining stages. The washing and filling steps were able to reduce the load of molds and yeasts. It is understood that the physical and functional adaptations implemented were important to increase the percentage of adequacy of the items, however, the persistence of shortcomings in the work process may compromise the quality of the final product, stressing the need for compliance with good practices. / O processamento de polpas constitui uma possibilidade para utilização de frutos por povos tradicionais como os quilombolas, inseridos em um contexto no qual frutos nativos são pouco consumidos. Para tanto, a qualidade higienicossanitária é fundamental na produção de um alimento seguro. O objetivo deste estudo foi avaliar o processamento e as condições higienicossanitárias de frutos e polpas de frutas em comunidade quilombola. A pesquisa foi realizada na Comunidade dos Remanescentes do Quilombo do Pombal, Goiás, Brasil. Para avaliação das condições físico-funcionais foi aplicada lista de verificação antes e após a realização de adequações estruturais, capacitações e implementação do Manual de Boas Práticas e Procedimentos Operacionais Padronizados. Os dados foram categorizados de acordo com o percentual de adequação. Foram realizadas análises microbiológicas de 32 amostras incluindo, frutos do Cerrado encontrados na comunidade e frutos e suas polpas em diferentes etapas do fluxograma. Foram elencados os perigos e Pontos Críticos de Controle no fluxograma de produção. O diagnóstico físico-funcional realizado indicou média de conformidade dos blocos de 30,18%, sendo classificado no Grupo 3, insatisfatório. Após intervenções, verificou-se 62,13% de conformidade, com classificação no Grupo 2, regular (p<0.05). O efeito da intervenção foi significativo (p<0,05) para cincos dos sete blocos avaliados. A análise microbiológica indicou que todos os frutos nativos coletados estão em conformidade com as recomendações da legislação sanitária vigente. Os frutos e polpas inseridos no fluxograma de processamento apresentaram-se de acordo aos padrões sanitários para contagem de coliformes termotolerantes e pesquisa de Salmonella spp, porém, a contagem de bolores e leveduras em oito polpas analisadas, indicou que cinco apresentaram contagem acima do estabelecido pela legislação. Os resultados sugerem contaminação do fruto já na colheita e a multiplicação destes micro-organismos nas demais etapas, sendo que as etapas de lavagem e envase foram capazes de reduzir a carga de bolores e leveduras. Entende-se que as adequações físico-funcionais implementadas foram importantes para o aumento do percentual de adequação dos itens, contudo, a persistência de falhas no processo de trabalho pode comprometer a qualidade do produto final, ressaltando a necessidade de adequação às boas práticas.

Salmonella Enteritidis (SE) é um enteropatógeno de grande preocupação para indústria de alimentos, principalmente de produtos que não podem ser submetidos a tratamento térmico, como as saladas à base de maionese, freqüentemente envolvidas em surtos de salmonelose. O uso de antimicrobianos naturais nesses produtos pode ser um método alternativo para o controle de SE. Assim, o objetivo deste trabalho foi avaliar o efeito antimicrobiano de óleo essencial de orégano, EDTA e nisina individualmente e a combinação de nisina com EDTA e nisina com óleo essencial de orégano no controle da multiplicação SE em salada de legumes com maionese. A atividade inibitória foi inicialmente avaliada in vitro e posteriormente em salada preparada com legumes, experimentalmente contaminados com SE, misturados à maionese, na concentração de 103 UFC/g, e armazenados em refrigeração (8°C) e em temperatura ambiente (30°C). Os resultados da avaliação in vitro indicaram que o OEO usado individualmente tem melhor efeito antimicrobiano contra SE do que quando empregado em combinação com nisina. Nem a nisina nem o EDTA, quando testados isoladamente ou combinados apresentaram efeito sobre SE. Na salada de legumes com maionese preparada artesanalmente, a presença de 0,2% de OEO resultou em redução na contagem de SE, constituindo-se em uma barreira adicional para a multiplicação do patógeno nesse produto. A análise sensorial da salada de legumes com maionese contendo 0,2% de OEO indicou que houve 95% de aceitação do aroma, 93% de aceitação do sabor e 74% de intenção de compra. A análise sensorial da maionese contendo 0,5% ou 1% de OEO indicou a inviabilidade de se aumentar a concentração do OEO na maionese como alternativa para melhorar o efeito antimicrobiano. / Salmonella Enteritidis (SE) is an enteropathogen relevant for the food industry, especially in foods that do not require heat treatment before consumption such as salads prepared with mayonnaise, frequently associated to outbreaks of salmonellosis. Natural antimicrobials can be used as an alternative procedure to control SE in these foods. The aim of the present study was to evaluate the effect of origanum essential oil (OEO), nisin and EDTA, used individually and in combination, on the growth of SE in mayonnaise and in mayonnaise-based legume salad. The inhibitory activity was evaluated in vitro, in home made mayonnaise and in mayonnaise-based legume salad experimentally contaminated with SE at 103 CFU/g, stored under refrigeration (8°C) and at room temperature (30°C). The results of the in vitro tests indicated that a better antimicrobial effect of OEO on SE was achieved when OEO was used individually than when used combined with nisin or EDTA. Nor nisin nor EDTA, used individually, presented any effect against SE. The presence of OEO in mayonnaise did not add any additional effect to the antimicrobial activity of the intrinsec parameters in this food. In mayonnaise-based legume salad, 0.2% of OEO caused a reduction in the counts of SE, and was an additional hurdle for the growth of the pathogen in this product. The sensorial testing of mayonnaise-based legume salad containing 0.2% OEO indicated an acceptance of 95% for the aroma and 93% for the taste and 74% of purchasing intention. The sensorial testing of mayonnaise containing 0.5% or 1% OEO indicated that the increase in the content of OEO to increase the antimicrobial activity is not feasible.

Bifidobacterium spp. são microrganismos probióticos que podem ser incorporados em produtos alimentícios. Entretanto, para que seus efeitos benéficos à saúde humana ocorram, é necessário que o número de células viáveis na hora do consumo seja, no mínimo, 106UFC/g. As bifidobactérias são sensíveis à elevada acidez e, por isso, torna-se necessária a busca por métodos que possam proteger a integridade da célula, sendo um deles a microencapsulação. Em uma primeira etapa do trabalho, Bifidobacterium lactis foi encapsulado em micropartículas de alginato e alginato modificado (alginatoquitosana, alginato-quitosana-sureteric e alginato-quitosana-acryl-eze) e sua sobrevivência e liberação das micropartículas em fluidos simulados do trato gastrintestinal foram mensuradas utilizando-se soluções tampão com pH 1,5, 5,6 e 7,5, na presença e na ausência de pepsina (3g/L), pancreatina (1g/L) e bile (10g/L). A liberação de células das micropartículas teve uma relação direta com o pH do tampão. A microencapsulação aumentou a taxa de sobrevivência de B. lactis, em comparação com células não encapsuladas, em soluções tampão com pH 1,5 sem a presença de enzimas. Em suco gástrico simulado com enzimas digestivas, por outro lado, foi observado que a pepsina proporcionou um efeito protetor sobre as células de B. lactis, e nesse caso, as taxas de sobrevivência do microrganismo estavam diretamente relacionadas com o grau de injúria das células. Em uma segunda etapa do trabalho, leites fermentados com Streptococcus salivarius ssp. thermophilus e Lactobacillus delbrueckii ssp. bulgaricus foram enriquecidos com culturas de Bifidobacterium lactis submetidas a quatro tratamentos diferentes: desidratação em temperatura ambiente, liofilização/congelamento, encapsulação em alginatoquitosana e encapsulação em alginato-quitosana-acryl-eze. A população sobrevivente de B. lactis foi determinada semanalmente no leite fermentado e também após tratamento simulando condições do trato gastrintestinal. Os resultados indicaram que na ausência de pepsina, as populações de B. lactis foram reduzidas drasticamente após o contato com tampão pH 1,5, não sendo possível a detecção de células viáveis livres ou encapsuladas após 120 minutos de teste. A presença de pepsina influenciou positivamente a recuperação de células viáveis de B. lactis em todas as condições testadas, mas as culturas na forma desidratada apresentaram melhores resultados que as culturas microencapsuladas ou liofilizadas. No caso do leite fermentado contendo as células desidratadas, a população de B. lactis, após o tratamento em suco gástrico com enzimas, foi superior à detectada no produto antes desse tratamento. Conclui-se que a microencapsulação não foi eficiente para proteger B. lactis em leite fermentado contra injúrias causadas pelo trato gastrintestinal simulado. / Bifidobacterium spp. are microorganisms that can be added to foods. However, the benefits for the human health occur when the numbers of viable cells in the moment of the consumption is at least 106CFU/g. Bifidobacteria are acid sensitive, and methods to protect cell integrity, such as microencapsulation, are needed. In the first part of the present study, Bifidobacterium lactis was encapsulated in microparticles of alginate and modified alginate (alginate-chitosan, alginate-chitosan-sureteric and alginate-chitosan-acryl-eze) and the survival and release from microparticles in simulated gastrointestinal conditions were measured, using buffers (pH 1.5, 5.6 and 7.5), in the absence and presence of pepsin (3g/L), pancreatin (1g/L) and bile. The release from microparticles presented a direct relationship with pH. When the pH was 1.5 and no enzyme was present, encapsulation improved the survival of B. lactis, when compared to free cells. However, pepsin had a protective effect on B. lactis, and the survival rate was directly related to the cells injury degree. In the second part of the study, fermented milk samples containing Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. Bulgaricus were supplemented with B. lactis submitted to four different treatments: dehydration at room temperature, freeze drying, encapsulation in alginate-chitosan and encapsulation in alginate-chitosaacryl-eze. The number of viable B. lactis cells in the fermented milk was determined weekly and also after treatment with simulated gastrointestinal conditions. Results indicated that in the absence of pepsin, the number of viable cells decreased significantly after contact with buffers (pH 1.5), and no viable cell was detected after 120 minutes. Pepsin improved the recovery of viable cells in the assayed gastric conditions, being the dehydrated cultures more resistant than other cultures. In fermented milk containing the dehydrated cells, the number of viable cells increased after treatment with simulated gastrointestinal fluids. Microencapsulation was not an effective procedure to protect B. lactis in fermented milk against injury caused by the simulated gastrointestinal tract.

Escherichia coli é um microrganismo presente no trato intestinal do homem e de animais de sangue quente, fazendo parte da microbiota, coexistindo sem causar danos ao hospedeiro. No entanto, algumas linhagens desse microrganismo podem ser patogênicas e causar doenças tanto ao homem como aos animais. E. coli produtoras de toxina de Shiga (STEC), consideradas patógenos de origem alimentar, podem causar desde diarréias brandas até severas e sanguinolentas a complicações graves, como colite hemorrágica (HC), síndrome urêmica hemolítica (HUS) e púrpura trombótica trombocitopênica (TTP). O gado é considerado um importante reservatório deste patógeno e a contaminação de seres humanos ocorre, na maioria das vezes, através do consumo de alimentos ou água contaminados. O presente trabalho teve como objetivos avaliar a ocorrência de E. coli O157:H7 e outras STEC em amostras de couro de animais bovinos e de suas respectivas carcaças, na etapa de pré-evisceração, e meia-carcaças, na etapa de pós-evisceração; identificar os genes que codificam para os fatores de virulência (stx1 , stx2, eaeA e ehxA) dos isolados obtidos; evidenciar cepas de E. coli O157:H7 através da pesquisa do gene uidA; identificar os sorotipos dos isolados; verificar a citotoxicidade dos isolados de STEC em células Vero e avaliar a sensibilidade a diferentes antibióticos. De 198 animais amostrados, sete (3,5%) apresentaram cepas de STEC. Em seis (3%) destes, STEC foi detectada no couro e em um (0,5%) foi isolada de meia-carcaça, não tendo sido detectada em amostras de carcaça. As 23 cepas isoladas do couro apresentaram o perfil stx2, eaeA, uidA e ehxA, podendo ser consideradas E. coli enterohemorrágica (EHEC), e a isolada de meia carcaça apresentou o perfil stx2, uidA e ehxA. Das 24 cepas isoladas, 13 (54,2%) pertenciam ao sorotipo O157:H7. Além deste sorotipo, foram isoladas cepas de outros sorotipos previamente descritos e associados a doenças humanas severas no Brasil e em outros países, como O174:H21, O6:H49, ONT:H7, ONT:H8 e OR:H10. Dos sete animais com cepas positivas para stx2e ehxA, cinco (71,4%) apresentaram cepas com atividade citotóxica em células Vero e um (14,2%) apresentou cepas positivas na avaliação da produção de entero-hemolisina. Com relação ao teste com antibióticos, quatro (16,7%) das 24 cepas testadas apresentaram resistência a um ou mais antibióticos, sendo três (12,5%) a estreptomicina e uma (4,2%) a estreptomicina e ampicilina. Diante destes resultados, pode-se dizer que a produção de entero-hemolisina e a pesquisa dos genes ehxA e uidA não demonstraram ser bons marcadores na pesquisa do sorotipo O157:H7. A presença de cepa de STEC na meia-carcaça alerta para a necessidade de vigilância da presença destes microrganismos, uma vez que eles poderiam contaminar o produto final, colocando em risco a saúde do consumidor. / Escherichia coli is a microorganism present in the intestinal tract of humans and warm-blood animals, being part of the normal microbiota and harmless to the host. However, some strains are able to cause human and animal infections. Shiga toxin-producing E. coli (STEC), regarded as foodborne pathogens, can cause since mild or severe and bloody diarrhea to major complications, such as hemorrhagic colitis (HC), hemolytic-uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP). Cattle are considered the main reservoir of this pathogen and the transmission to humans happens, most of the times, due to the consumption of contaminated food or water. The aim of the present research was to determine the prevalence of E. coli O157:H7 and other STEC on hide samples of beef cattle and on their corresponding carcasses, sampled prior to evisceration, and half-carcasses, sampled after evisceration; identity the genes that code for the virulence factors (stx1, stx2, eaeA e ehxA) of the isolates; detect E. coli O157:H7 strains using the gene uidA as epidemiological marker; identify the serotypes of the STEC isolates; verify the citotoxicity of the isolates in Vero cells and evaluate their resistance to different antibiotics. From 198 animals sampled, seven (3.5%) carried STEC strains. In six (3%) of them, STEC was detected on hide and in one (0.5%) it was isolated from half-carcass. The 23 strains isolated from hide presented the profile stx2, eaeA, uidA e ehxA, and were regarded as enterohemorrhagic Escherichia coli (EHEC), and the one isolated from half-carcass presented the profile stx2, uidA e ehxA. From the 24 isolated strains, 13 (54.2%) belonged to the serotype O157:H7. Besides this serotype, other strains belonging to serotypes that have been previously described and associated with severe human infections in Brazil and other countries, such as O174:H21 , O6:H49, ONT:H7, ONT:H8 and OR:H10, were isolated. From seven animals with strains harboring stx2, and ehxA, five (71.4%) presented verocytotoxigenic strains and one (14.2%) presented enterohemolisin producing strains. Regarding the antibiotics tested, four (16.7%) of the 24 isolated strains were resistant to some antibiotic, being three (12.5%) to streptomycin and one (4.2%) to streptomycin and ampicilin. Faced with these results, the production of enterohemolisin and the search of the genes ehxA and uidA can not be considered good epidemiological markers for the serotype O157:H7. The isolation of STEC strain from the half-carcass alerts for the need of surveillance on the presence of these microorganisms, since they may contaminate the final product, representing a risk to consumers health.

Agroterrorism is the deliberate introduction of a plant or animal disease with thegoal of causing fear, economic instability, illness, or death. After the 2002 terroristattacks on the World Trade Center, the security of the food supply is of increasingconcern to the United States. A major incidence of agroterrorism or food tampering would have far reaching impacts on the economy and public health. The first objective of this project was to determine knowledge and concern of agroterrorism in meat processing facilities in Kentucky, and to determine knowledge and concern of food tampering and food defense in food service establishments in Warren County, Kentucky. The second objective was to determine security strategies that were being implemented by these facilities. Two separate surveys, one for meat processors and the other for food service establishments, were designed to meet these objectives. An observational study was conducted for meat processing facilities. It was found that these facilities were generally unconcerned with agroterrorism, although a reasonable amount of security implementations were in place at these facilities. A statistical comparison between restaurants and non-restaurant food service establishments, such as schools, hospitals, and hotels, was performed. Both types of food service establishments expressed little concern about a food tampering event. Non- restaurant food service establishments were slightly more concerned than restaurants about both food tampering and food defense.

Thesis (MScFoodSc)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: The regular consumption of probiotics is becoming a recognized trend in the food industry due to several reported health benefits. A probiotic is defined as a live microbial feed supplement that beneficially affects the host animal by improving its intestinal microbial balance. A wide variety of probiotic food products are available on the South African market and comprise an assortment of fermented milks, as well as lyophilized preparations in tablet or capsule form. Strains of Lactobacillus acidophilus and Bifidobacterium species are mostly used as probiotic microbes in the industry due to their health enhancing effect. The survival of sensitive probiotic microbial species in food matrices are influenced by various factors such as oxygen concentration, pH levels and manufacturing and storage conditions. These should be considered and monitored as the South African food and health regulations stipulate that probiotic microbes should be present at a concentration of 10⁶ cfu.ml ̄ ¹' in order to exert a beneficial effect. Some health benefits are also correlated to specific microbial species and strains and these factors have resulted in the need for the rapid and accurate identification of probiotic microbes present in food products. The probiotic microbes present in probiotic yoghurts and supplements have in the past been identified using traditional methods such as growth on selective media, morphological, physiological and biochemical characteristics. However, even some of the most sophisticated cultural-dependant techniques are not always sufficient for the identification and classification of especially Bifidobacterium, as well as closely related Lactobacillus species. Molecular techniques are more often employed for the rapid and accurate detection, identification and characterization of microbial species present in food products. The aim of this study was to detect and identify the probiotic species present in various commercial South African yoghurts and lyophilized preparations using peR-based DGGE analysis. A 200 bp fragment of the V2-V3 region of the 16S rRNA gene was amplified and the peR fragments were resolved by DGGE. The unique fingerprints obtained for each product were compared to two reference markers A and B in order to identify the bands present. The results obtained were verified by species-specific peR, as well as sequence analyses of bands that could not be identified when compared to the reference markers. Only 54.5% of the South African probiotic yoghurts that were tested did contain all the microbial species as were mentioned on the labels of these products, compared to merely one third (33.3%) of the lyophilized probiotic food supplements. Some Bifidobacterium species were incorrectly identified according to some product labels, while other products contained various microbes that were not mentioned on the label. Sequence analysis confirmed the presence of a potential pathogenic Streptococcus species in one of the yoghurt products and in some instances the probiotic species claimed on the labels were non-scientific and misleading. The data obtained in this study showed that the various South African probiotic products tested were of poor quality and did not conform to the South African regulations. peR-based DGGE analysis proofed to be a valuable approach for the rapid and accurate detection and identification of the microbial species present in South African probiotic products. This could help with future implementation of quality control procedures in order to ensure a reliable and safe probiotic product to the consumer. / AFRIKAANSE OPSOMMING: Die gereelde inname van probiotiese produkte is besig om In erkende tendens in die voedselindustrie te word, as gevolg van verskeie gesondheidsvoordele wat daaraan gekoppel word. In Probiotika word gedefinieer as In voedingsaanvulling wat uit lewendige mikrobes bestaan en wat In voordelige effek op mens of dier het deur In optimale mikrobiese balans in die ingewande te handhaaf. In Wye verskeidenheid probiotiese voedselprodukte is tans beskikbaar op die Suid- Afrikaanse mark. Hierdie bestaan hoofsaaklik uit verskeie gefermenteerde melkprodukte asook 'n reeks tablette en kapsules wat probiotiese mikrobes in gevriesdroogde vorm bevat. Lactobacillus acidophilus tipes en Bifidobacterium spesies word die algemeenste in die voedselindustrie gebruik aangesien hierdie spesifieke mikrobes bekend is om goeie gesondheid te bevorder. Die oorlewing van sensitiewe probiotiese mikrobiese spesies in voedsel matrikse word beïnvloed deur faktore soos suurstof konsentrasie, pH-vlakke en vervaardigings- en opbergings kondisies. Hierdie faktore moet in aanmerking geneem word en verkieslik gemonitor word aangesien die Suid-Afrikaanse voedsel en gesondheids regulasies stipuleer dat probiotiese mikrobes teen In konsentrasie van 10⁶ kolonie vormende eenhede per ml teenwoordig moet wees om In voordelige effek te toon. Sommige gesondheidsvoordele word direk gekoppel aan spesifieke mikrobiese spesies en spesie-tipes. Hierdie faktore het gelei tot In groot aanvraag na vinnige en akkurate metodes vir die identifikasie van probioties mikrobes in voedselprodukte. Die probiotiese mikrobes teenwoordig in probiotiese joghurts en ook die gevriesdroogde vorms in tablette en kapsules, was al geïdentifiseer deur gebruik te maak van tradisionele metodes soos groei op selektiewe media, morfologiese, fisiologiese en biochemiese eienskappe. Selfs van die mees gesofistikeerde kultuur-afhanklike tegnieke is egter nie altyd voldoende vir die identifikasie en klassifikasie van veral Bifidobacterium en na-verwante Lactobacillus spesies nie. Molekulêre metodes word dikwels aangewend vir die vinnige en akkurate deteksie, identifikasie en karakterisering van mikrobes teenwoordig in voedselprodukte. Die doel van hierdie studie was om die probiotiese mikrobes teenwoordig in verskeie Suid-Afrikaanse joghurts en gevriesdroogde aanvullings, te identifiseer deur gebruik te maak van polimerase kettingreaksie (PKR)-gebaseerde denaturerende gradiënt jelelektroforese (DGGE) analise. 'n PKR fragment van 200 bp van die V2-V3 gedeelte van die 16S ribosomale RNS (rRNS) geen is geamplifiseer, en die PKR fragmente is geskei met behulp van DGGE. Die unieke vingerafdrukke wat verkry is vir elke produk is teen twee verwysings merkers A en B vegelyk om die bande teenwoordig in die profiele te identifiseer. Die resultate is bevestig deur spesies-spesifieke PKR en ook deur die ketting volgordes van die DNS fragmente te bepaal wat nie geïdentifiseer kon word deur vergelyking met die verwysings merkers nie. Slegs 54.5% van die Suid-Afrikaanse probiotiese joghurts wat getoets is het al die mikrobiese spesies bevat soos aangedui was op die etikette van hierdie produkte, teenoor slegs 'n derde (33.3%) van die gevriesdroogde voedingsaanvullings. Sekere Bifidobacterium spesies is verkeerd geïdentifiseer op sommige van die produk etikette, terwyl ander produkte verskeie mikrobes bevat het wat nie op die etiket aangedui was nie. 'n Potensiële patogeniese Streptococcus spesie is in een van die joghurt produkte gevind soos bevestig deur DNS kettingvolgorde bepalings. In sommige gevalle was die probiotiese spesienaam wat aangedui is op die etiket onwetenskaplik en misleidend. Die resultate wat uit hierdie studie verkry is dui aan dat die Suid-Afrikaanse probiotiese produkte wat getoets is van 'n swak gehalte is en nie aan die Suid- Afrikaanse regulasies voldoen nie. Daar is getoon dat PKR-gebaseerde DGGE analise 'n waardevolle tegniek kan wees vir die akkurate deteksie en identifisering van die mikrobiese spesies teenwoordig in probiotiese produkte. Dit kan help met die toekomstige implementering van kwaliteitskontrolerings prosedures om 'n mikrobiologiese betroubare en veilige produk aan die verbruiker te verseker.

Phthalate esters (PEs) belong to a class of organic compounds used as plasticisers in plastic materials such as polyvinyl chloride (PVC), polypropylene (PP), polyethylene terephthalate (PET) and so on, including those used in the food packaging industry. Phthalate plasticisers are not chemically bound to plastic materials and hence, migrate into items such as foodstuffs they house. The study aimed at investigating the prevalence of selected phthalate esters from plastic wrappers into food as well as the presence of food and/or pathogenic microorganisms. Plastic-wrapped cheese, vienna sausages and polony samples purchased from commercial stores in the four regions of Pretoria (Tswane), South Africa, were analysed for the presence of plasticisers; di-2-ethylhexyl adipate (DEHA), di-n-butyl phthalate (DnBP), benzyl-butyl phthalate (BBP), di-butyl phthalate (DBP) and dimethyl phthalate (DMP). Soxhlet extraction using hexane with florisil column cleanup was carried out. Analysis of PEs was by Gas Chromatography-Flame Ionization Detection (GC-FID). Microbiological investigations were performed using standard methods. The concentrations of PEs detected in food samples ranged from below detection limit (bdl) to 4.7003 μg/kg. However, DBP, DMP and BBP were predominantly present with more PEs detected in cheese compared to polony and vienna. In polony samples, DBP levels ranged from 0.0412 to 0.611μg/kg, in cheese, ranged from 0.049 to 0.256 μg/kg and in vienna DBP ranged from 0.074 to 0.209 μg/kg. The phthalate DMP ranged from 0.072 to 4.700 μg/kg in cheese, 0.056 to 0.241 μg/kg in polony and 0.092 to 0.816 μg/kg in vienna. The DEHA detected in cheese and polony was 0.120 μg/kg and 0.075 μg/kg respectively and no DEHA was detected in vienna sausages. For microbiological analysis, the total microbial activity (TMA) ranged from 6.8 x 104 to 1.03 x 108 cfu/g; coliforms ranged from no growth to 2.62 x 106 cfu/g; yeast ranged from no growth to 1.49 x 107 cfu/g; and mould ranged from no growth to 9.2 x 104 cfu/g. The results revealed that microbial activity was high in each sample type but revealed the absence of pathogens. Results revealed incidences of PEs in foods wrapped or packaged in plastics, which gave cause for concern and showed the need for proper monitoring and inspection of the levels of organo-microbial entities in the South African food wrapped in plastic wrappers. / Environmental Sciences / M.Sc. (Environmental Science)