Dissertations / Theses on the topic 'Food microbiology (Analysis)'

A gas chromatographic method for the routine analysis of volatile and semi-volatile sulphur compounds in beer was developed. Various selective and specific detectors, capillary columns and methods of sample preparation were compared. The combination offering the best sensitivity and stability consisted of solid phase microextraction (SPME) as the sample preparation step; a combined polar/non-polar chromatographic column; and a pulsed flame photometric detector (PFPD). All parameters were optimised to achieve maximum sensitivity. The system was linear for the range of sulphur compound concentrations found in beer, and displayed good reproducibility. The calibrated SPME-GC-PFPD system was used to analyse several different varieties of beer from a range of breweries, and to investigate the change in the concentrations of sulphur compounds when beer is subjected to illumination.

The practical work described in this thesis falls into four distinct subject areas: HPLC, tea liquor fractionation, mass spectrometry, and NMR and IR. The practical work, is therefore written in four chapters, each chapter having its own introduction, results and discussion, and experimental sections. The work is concerned with the analysis of black tea liquor, its aim being to provide an analytical input into a larger project, being carried at NRI (Natural Resources Institute), on the statistical evaluation of tea quality.

Recent epidemiological studies have suggested that the development and progression of several chronic diseases may be initiated or augmented by oxidative stress. Reactive oxygen species and reactive nitrogen species react readily with and can damage nucleic acids, proteins, and lipids. While biological systems are equipped antioxidant defenses to cope with oxidative stress, oxidative damage may still occur when oxidative stress overwhelms antioxidant defenses. This damage, if left unchecked, may lead to a variety of degenerative diseases, including heart disease, Alzheimer's Disease, Parkinson's Disease and cancer. Several assays have been designed to describe the antioxidant activity of various phytochemicals, vitamins, and other compounds. The ORAC and TOSC assays have emerged as industry standards for measuring antioxidant activity due to their high reliability and sensitivity. Until recently, however, little has been done to assess the relative correlation between these two assays. Furthermore, no assay has been developed to measure changes in antioxidant activities of cells in response to oxidative stress. The current work investigates the correlation between measured antioxidant activities of samples in the both the ORAC and TOSC assays. Recent antioxidant research also focuses on relating chemical structure to antioxidant activity. Previous research in this area has included a broad range of chemical groups, but no study has attempted to formulate a structure-function framework that has applicability to compounds of any group. The current work uses amino acids as a simplest-case model for studying the relationships between chemical structure and antioxidant activity. One particular area of emerging research has centered around comparing organic and conventionally grown food products. The impetus of these investigations lies in claims made by organic supporting groups that these food products are generally more beneficial than their conventional counterparts. Despite the rapid rise in popularity of organic foods, there remains a dearth of research investigating these claims. The current work compares the antioxidant activities of organic and conventionally grown blueberries and apples.

Thesis (Masters Diploma (Food Technology))--Cape Technikon, Cape Town,1993
This study is based on the Hazard Analysis in the Wet Milling of maize for the production of starch at the Bellville plant of African Products. Wet milling of maize is a highly specific and completely integrated system developed to separate the major components of the kernel as completely as possible. Many microbiological problems existed in the process at this plant which could not be solved over the years. Microbial analyses were done throughout the plant and high microbial counts were obtained at various sampling points. In applying HACCP, the following major hazards were identified: The presence of Faecal Streptococci, Sraphylococcus aureus, Bacillus cereus, Faecal coliforms, Fusarium, Dip/odia, Aspergillus, Penicillium and various Yeast strains. The follOWing Critical Control Points (CCP's) were identified in the wet milling process: Maize trucks, in-process water, steeping, storage tanks, Reineveld, wet mlxmg boxes, Laidlaw, drying and bagging off point. The follOWing were done as part of the HACCP plan: i) modifications of the plant were suggested, ii) different sanitation programmes were evaluated, iii) monitoring of cep's, and iv) training of personnel. In general, a regular sanitation programme need to be exercised in the wet-milling plant to prevent a build up of microbial populations at various sampling points. High S02 levels can be maintained throughout the plant to achieve this. The final starch will then be used for Industrial starch. Criteria to monitor the CCP's were suggested. Hazard Analysis is an effective method to improve the quality of the final product.

Matthew R. Borglin This thesis demonstrates efficacy of Octenyl Succinic Anhydride (OSA), as a biofilm sanitizer. Biofilms allow bacteria to adhere to solid surfaces with the use of excreted polymeric compounds. For example, surfaces found in food production or processing facilities such as the interior of a raw milk holding tank, are some of the most susceptible to biofilm contamination. When present, biofilms can cause a variety of negative effects, which include; reduction of product shelf life, corrosion, and outbreaks of foodborne illnesses. The close association of biofilms with the majority of foodborne illness cases led the US Environmental Protection Agency (EPA) to create a new category of sanitizer specifically designed for treatment of mature biofilms. The efficacy of sanitizers in this new regulatory category is determined by the EPA protocols MB-19 and MB-20. The EPA’s protocols outline methods for cultivating, treating, and measuring effects on Pseudomonas aeruginosa biofilms in a continuous flow stir bar bioreactor. Biofilm modification by OSA was verified by the presence of octenyl esters on OSA treated biofilms with single point Raman spectrophotometry. OSA modified biofilm’s antimicrobial properties were first investigated with crystal violet staining in 96-well microtiter plates with inconclusive results. However, effective antimicrobial properties where apparent when using the CDC Biofilm Reactor. OSA treatments consistently returned a 6-log CFU/coupon reduction in biomass compared to controls. Inhibition of planktonic and/or biofilm regrowth was demonstrated using the 96-well plate methodology. This thesis demonstrated the effectiveness of OSA chemical esterification reaction as a biofilm treatment. In doing so, this work suggests a new approach for biofilm remediation by chemically modifying the structural components of biofilm.

A demanda por frutas e hortaliças frescas, associada à necessidade de maior praticidade da vida atual, está causando um aumento no interesse, por parte dos consumidores, nos produtos minimamente processados (MP). Processamento mínimo inclui as operações de lavagem, corte, descascamento e embalagem do produto. Entre os microrganismos patogênicos que, potencialmente, podem ser transmitidos por vegetais MP citam-se: Listeria monocytogenes, Escherichia coli O157:H7 e Salmonella sp. A pesquisa destes microrganismos é usualmente demorada mas, a cada dia, novos métodos para detecção rápida de patógenos em alimentos são lançados no mercado. Dentre estes métodos, aqueles que empregam ferramentas moleculares têm se tornado mais populares, cabendo destacar os que empregam a reação de polimerização em cadeia (PCR). Para a pesquisa de Listeria monocytogenes existe no mercado o sistema automatizado BAX®System que permite a detecção de L. monocytogenes em, no máximo, 54 h. Neste estudo, buscou-se avaliar a microbiota de vegetais folhosos MP além do emprego do sistema BAX® para a detecção de L. monocytogenes nestes produtos. Foram examinadas, no período de março a julho de 2003, 181 amostras de saladas MP coletadas em diferentes estabelecimentos comerciais no município de São Paulo, SP. Em 133 amostras foram feitas determinações das populações de coliformes totais e fecais, Enterobacteriaceae, microrganismos psicrotróficos aeróbios e pesquisa de Salmonella sp. L. monocytogenes foi pesquisada nas 181 amostras empregando-se o sistema BAX® e, paralelamente, a semeadura do caldo de enriquecimento em placas contendo ágar Palcam e Oxford, com a identificação das colônias suspeitas através de testes bioquímicos tradicionais. Das 133 amostras, 51% apresentaram populações de microrganismos psicrotróficos aeróbios > 106 UFC/g e 42% apresentaram populações de Enterobacteriaceae entre 105 – 106 UFC/g. Coliformes fecais estiveram presente em populações superiores a 102 UFC/g em 97 amostras (73%) e Salmonella foi detectada em 4 amostras (3%). L. monocytogenes estava presente em 1 (0.6%) amostra de espinafre das 181 amostras examinadas, tendo sido detectada, simultaneamente, por ambos os métodos empregados. As outras espécies de Listeria encontradas, empregando-se a semeadura em placa foram: L. welshimeri (1 amostra de alface mimosa) e L. innocua (2 amostras de agrião). Os resultados indicam que grande parte dos vegetais MP examinados, apresentaram qualidade microbiológica deficiente e podem ser veículos de patógenos como a Salmonella. O BAX®System é de grande utilidade para as análises de vegetais MP, que permite a obtenção de resultados mais rapidamente que o método tradicional, sem perda na sensibilidade e na especificidade.
The increasing demand for fresh fruits and vegetables, associated with the desire of convenient goodies, is causing an expansion on the market share of minimally processed products (MP). Minimal processing includes operations such as washing, cutting, peeling and packaging of the product. Amongst pathogenic microorganisms that can be transmitted by MP vegetables are: Listeria monocytogenes (Lm), Escherichia coli O157:H7 and Salmonella sp. Searching for these microorganisms is labor intense and time-consuming, however new methods for fast detection of pathogens are commercially available. Methods employing molecular technology are becoming more popular and the polymerase chain reaction (PCR) is now a good choice. There is an automatized PCR system (BAX®System) that can be used for Lm detection in up to 54 h. The aims of this study was to evaluate the microflora of MP vegetables and to evaluate the effectiveness of the BAX®System for screening Lm on those products. From March to July 2003, 181 samples of MP salads were collected at retail level in the city of São Paulo, SP. Total and faecal coliforms, Enterobacteriaceae, psychrotrophic microorganisms enumeration and Salmonella evaluation were conducted in 133 samples. L. monocytogenes was assessed in 181 samples using the BAX®System and also by plating the enrichment broth onto palcam and Oxford agars. Suspected colonies of Listeria were submited to classical biochemical tests. Population of psychrotrophic microorganisms >106 CFU/g was observed in 51% of the 133 samples and Enterobacteriaceae population between 105 - 106 CFU/g was in 42%. 97 samples (73%) showed population of faecal coliforms >102 CFU/g (Brazilian standard) and Salmonella was detected in 4 samples (3%). L. monocytogenes was detected in 1 spinach sample (0,6%) out of the 181 examined MP vegetables. This positive sample was simultaneously detected by both methods. The other Listeria species identified by plating were L. welshimeri (1 sample of curly lettuce) and L. innocua (2 watercress samples). The results indicate that the MP vegetables had poor microbiological quality and could be vehicle of pathogens such as Salmonella. BAX®System showed good specificity and sensitivity when used for vegetable analysis and was easier to perform and faster than the classical method.

Water is the most consumed foodstuff around the world. Therefore it is very important to analyze possible faecal contamination of water, and Enterococci are an indicator for that. They are also used as an indicator for possible faecal contamination in food. Normally you find Enterococci in the intestines of humans and animals, but Enterococci can also give infections like urinary tract infection.

In this study, varying number of colonies and colours of Enterococci on different media were evaluated. The purpose was to investigate if three different methods would give the same numbers of colonies. Another interesting perspective was to investigate if one medium could be used for two methods. Membrane filter techniques and surface spreading techniques were used to detect Enterococci. These methods were compared with a most probable number method.

None of the strains showed an optimal result on all media, however one medium, ChromoCult, showed good results for all investigated strains.

Use of the bioluminometric ATP assay was evaluated for estimating total bacterial counts in ground beef. Minimum sensitivity was found to be 10⁶ cfu/g using a double filtration procedure for sample preparation. Although ATP content per cfu decreased approximately 10 fold during storage, correlation of total aerobic plate count (APC) with microbial ATP content was 0.96. Selective non-microbial ATP extraction with ATPase treatment was evaluated for use in conjunction with the double filtration procedure to increase assay sensitivity. The new method was effective for removing additional non-microbial ATP without reducing ATP in bacteria. Estimated APC values were generally accurate to within ±0.50 log for ground beef samples above the detection limit of 5 x 10⁴ cfu/g. ATPase treatment increased sensitivity of the ATP assay and APC estimation by about 1 log while increasing assay time by 40 minutes, for a total of 60 minutes for 4 samples assayed in triplicate. The ATP assay was evaluated for use with ground beef patties inoculated with mixed ground beef spoilage flora, Pseudomonas, or Lactobacillus and stored at 2°C or 10°C using oxygen permeable or impermeable (vacuum) packaging. Excellent correlation (r²=0.95) was obtained for each inoculum and storage condition over the range of 5 x 10⁴ to 1 x 10⁹ cfu/g, when estimated APC values were compared with experimentally observed APC values. Usefulness of the ATP assay for estimating APC values of frozen ground beef was evaluated. Retail ground beef and Lactobacillus- and Pseudomonas-inoculated beef were frozen and thawed at different rates and examined for APC and microbial ATP content. Results indicated that, although freezing and thawing lowered numbers of Pseudomonas, APC values and microbial ATP content closely correlated. APC estimates were generally accurate to within 1/2 log. The importance of using an ATP assay standard to correct for variable enzyme activity and presence of quenching factors was demonstrated, and improved formulae were developed for optimum assay standard use. Alternate regression methods were evaluated for estimation of APC values but did not yield enhanced accuracy. Only one regression equation was needed for estimating APC values of ground beef containing different types of bacteria stored in various ways. Therefore, little knowledge of ground beef history is needed in order to rapidly and accurately estimate microbial numbers in ground beef using the bioluminometric ATP assay.
Ph. D.

Thesis (Ph.D.) - University of Glasgow, 2008.
Ph.D. thesis submitted to the Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, 2008. Includes bibliographical references. Print version also available.

La validité des contrôles microbiologiques, réalisés dans l'objectif d'assurer la sécurité sanitaire des aliments, nécessite notamment l'obtention de résultats d'analyse fiables. La fiabilité des résultats implique l'utilisation de méthodes validées, mises en œuvre par un laboratoire compétent. Les essais inter-laboratoires permettent de s'assurer, du moins en partie, du respect de ces deux conditions. Cependant, en raison de limites expérimentales, ces essais ne sont pas aussi largement pratiqués dans le domaine de la microbiologie des aliments qu'ils ne le sont dans d'autres domaines analytiques. Dans un premier temps, une revue des documents de référence permet d'établir un état des lieux. Cette revue concerne les trois objectifs que l'on peut assigner à des essais interlaboratoires, à savoir l'évaluation de méthodes d'analyse, celle des laboratoires, et la caractérisation de matériaux de référence. Les documents de portée générale, puis ceux spécifiques de l'analyse des aliments, sont pris en compte, et leur degré d'applicabilité à l'analyse microbiologique des aliments est envisagé. Les référentiels et pratiques propres au domaine d'intérêt traité sont finalement présentés, et les déviations par rapport aux documents généraux analysées. Sur cette base, sont présentées les conditions de mise en œuvre de deux types d'essais interlaboratoires, soit la validation de méthodes dans le cadre d'un projet européen du 4ème Programme Cadre de Recherche & Développement d'une part, et l'évaluation de laboratoires par le biais d'essais d'aptitude pour les Laboratoires Nationaux de Référence sur le lait d'autre part. Les difficultés relatives au protocole expérimental, et liées aux spécificités de la microbiologie, sont mises en exergue. Les modes d'exploitation des résultats, en fonction des objectifs et de la nature, qualitative ou quantitative, de la détermination, sont expliqués. En ce qui concerne la caractérisation de la performance des méthodes d'analyse, l'utilisation de statistiques robustes pour estimer la fidélité des méthodes quantitatives est discutée, ainsi que la façon de caractériser la fidélité comme la justesse des méthodes qualitatives. Sur ces aspects, des perspectives d'amélioration sont envisagées. L'intérêt de l'organisation des essais inter-laboratoires en microbiologie des aliments est ensuite abordé. Celui-ci réside dans l'utilisation que l'on peut faire de ces essais comme éléments incontournables de validation d'une méthode d'analyse et d'évaluation d'un laboratoire afin, d'une part, de crédibiliser ou d'améliorer les méthodes d'analyse normalisées au niveau international, et d'autre part d'estimer l'incertitude de mesure attachée aux résultats d'analyse. Quant aux limites de ces essais, essentiellement d'ordre expérimental, elles tiennent surtout à la nature vivante de l'analyte, et concernent des questions de représentativité.

Amostras de açúcar mascavo disponíveis em diferentes estabelecimentos comerciais no município de Araras - SP foram adquiridas para análises. No total foram coletados 49 pacotes de açúcar durante um período de quatro meses. As análises foram realizadas com o objetivo de avaliar a qualidade desse produto oferecido ao consumidor. As análises microbiológicas foram realizadas considerando a pesquisa dos seguintes microrganismos: bactérias mesófilas, bolores e leveduras, esporos de bactérias termófilas flat-sour, esporos de bactérias termófilas anaeróbias produtoras de H2S, esporos de bactérias termófilas anaeróbias não produtoras de H2S, Salmonella, coliformes totais e termotolerantes. Ainda foram determinadas a umidade (%) e atividade de água das amostras. Os resultados das análises apresentaram variações nas concentrações dos microrganismos estudados, cabendo às bactérias mesófilas e aos esporos de termófilas flat-sour as contagens mais elevadas, seguidos por bolores e leveduras. Todas as marcas apresentaram ausência de Salmonella, coliformes totais e termotolerantes, atendendo a legislação brasileira. Em relação ao padrão da National Canners Association apenas as marcas A, G, H e J estavam adequadas a essas exigências, as demais estavam em desacordo. Para o padrão da International Commission for Uniform Methods of Sugar Analysis (ICUMSA), nenhuma marca obteve resultado satisfatório em relação à qualidade microbiológica. Os teores de umidade variaram de 1,94 a 3,63% e os valores de atividade de água variaram de 0,55 a 0,64, havendo diferença significativa ao nível de 5% de significância. Pelos resultados obtidos considera-se que as marcas comercializadas estão de acordo com a legislação vigente no país, porém a maioria das marcas não atende aos padrões internacionais.
Samples of brown sugar available in different establishments in the city of Araras - SP were acquired for analysis. In the total 49 packages of sugar were collected during a period of four months. The analyses were performed to evaluate the quality of that product for the consumer. Microbiological tests were performed to search the following microorganisms: mesophilic bacteria, yeasts and molds, thermophilic flat-sour spores, thermophilic anaerobic H2S-producing spores, thermophilic anaerobic non-producing H2S spores, Salmonella and total and thermotolerant coliforms. Also humidity (%) and water activity were determined for the samples. The analysis results presented variations in the concentrations of microorganisms studied, being the mesophilic bacteria and thermophilic \"flat-sour\" spores the ones with highest scores, followed by yeasts and molds. All brands showed absence of Salmonella and total and thermotolerant coliforms, in accordance with the Brazilian legislation. Regarding the National Canners Association standards, only brands A, G, H and J were suited to these demands, the others being in disagreement. For the International Commission for Uniform Methods of Sugar Analysis (ICUMSA), standards no brands obtained satisfactory results in relation to microbiological quality. The moisture content ranged from 1.94 to 3.63% and the values of water activity ranged from 0.55 to 0.64, significant difference at 5% significance level. By the results it is considered that the brands are marketed under the law of the country, but most brands do not meet international standards. Humidity rates ranged from 1.94 to 3.63% and water activity ranged from 0.55 to 0.64, with significant difference at 5% significance level. By the results obtained it is considered that all brands sold are in accordance to the laws of the country, but most brands do not meet the international standards.

O queijo Minas frescal é um produto muito perecível e de curta duração. É necessário boas práticas de higienização durante sua produção e condições de armazenamento adequado para evitar a contaminação de microrganismos patogênicos e deteriorantes. Este trabalho teve por objetivo avaliar a vida útil do queijo Minas frescal durante o período de 30 dias armazenado a 4°C, através de análises sensoriais, microbiológicas e físico-químicas. Foram selecionadas seis marcas de queijo Minas frescal com registro no Serviço de Inspeção Federal (SIF), comercializados em supermercados de Piracicaba/SP até ser encontrado um produto dentro dos padrões da legislação vigente (BRASIL, 2001). O queijo selecionado foi adquirido direto do próprio lacticínio. As análises foram realizadas nos períodos de 1, 10, 20 e 30 dias após a fabricação do produto em três diferentes lotes denominados A, B e C. A contagem da população de microrganismos psicrotróficos totais, psicrotróficas proteolíticos, psicrotróficas lipolíticos, bactérias lácticas assim como de bactérias mesófilas totais apresentaram constante aumento mostrando crescimento de 8,50; 8,04; 8,30; 6,05 e 7,7 log UFC/g, respectivamente. Apenas do 30º dia do lote C o produto apresentou-se impróprio para o consumo com 1,7x104 NMP/g de Escherichia coli. Observou-se a redução do pH de 6,66 a 5,85 (0,81) e o aumento do ácido láctico de 0,044 a 0,07% (0,026%). O teor de gordura do queijo foi de 21,3% não sendo observado a sua redução. A quantidade de ácidos graxos livres foram determinados pela lipólise a qual aumentou em 0,22 mg KOH/g de gordura juntamente com a evolução dos microrganismos lipolíticos. A proteína variou de 21,30% a 22,10% sem diferença estatística significativa. A análise sensorial realizada pelo teste de aceitação através da escala hedônia teve entre 6,5 a 7,5 como pontuação o que indica que os julgadores gostaram ligeiramente e gostaram moderadamente do produto respectivamente durante o período de 30 dias. Conclui-se que o desenvolvimento das bactérias mesófilas, psicrotróficas totais, psicrotróficas proteolíticas, psicrotróficas lipolíticas e bactérias lácticas tem crescimento constante no queijo mesmo sob refrigeração levando este a se deteriorar com o passar do tempo. Da mesma forma o aumento significativo da acidez e de ácidos graxos livres. Apesar do aumento dos microrganismos e as alterações químicas o produto não teve mudanças sensoriais quanto aos atributos aparência, cor, odor, sabor e textura perante os dias de armazenamento. Uma vez que o queijo seja produzido sob ótimas condições higiênico sanitárias, ele é capaz de atingir o período de 30 dias de vida útil quando mantido a 4° C.
The Minas fresh cheese is very perishable product and its a short shelf life. Good practical of hygienic condition is necessary during its production and adjusted storage to prevent the proliferation of pathogenic and damage microorganism. This work had the objective evaluating the shelf life of the Minas fresh cheese for 30 days storaged at 4°C, through sensorial analyses, microbiological and physic-chemistries. Six marks of Minas fresh cheese commercialized in supermarkets in city Piracicaba/SP were selected, those are registered in Serviço de Inspeção Federal (SIF), but just one of them is inside of the standards of the current law (BRASIL, 2001). The select cheese was acquire direct of the Factory Dairy. The analyses were carried out in the periods of 1, 10, 20 and 30 days after the manufacturing of the product in the three different lots named A, B and C. The counting of the population of psychrotrophic microorganism total, psychrotrophic proteolytic, psychrotrophic lipolytic, lactic acidy bacteria as well as of mesophilic bacteria pesent constant increase showing growth of 8,50, 8,04, 8,30, 6,05 and 7,7 log UFC/g, respectively. On the 30th day of the lot C the product showed improper for the consumption with 1,7x104 NMP/g de Escherichia coli. It was observed a reduction of pH of 6,66 the 5,85 (0,81) and the increase of the lactic acidy of 0,044 the 0,07% (0,026%). The content of the cheese fat was 21,3% therefore its reduction wasn't observed. The amount of free fatty acid was determined by lipolysis which increased in 0,22 mg KOH/g of fat together with the evolution of the lipolytics microorganisms. The protein varied from 21,30% to 22,10% without difference significant statistics. The sensorial analysis carried through by the test of acceptance out the hedônic scale had punctuation between 6,5 the 7,5 what it indicates that the judges liked slightly and moderately respectively during the period of 30 days. The conclusion is that the development of the mesophilic bacteria, total psychrotrophic microorganism total, psychrotrophic proteolytic, psychrotrophic lipolytic and lactic acidy bacteria has constant growth in the cheese under refrigeration taking it this if to spoil with passing of the time. In the same way the significant increase of the acidity and free fatty acid. Despite of the increase of the microorganisms and the chemical alterations the product did not have sensorial changes how much to the attributes appearance, color, odor, flavor and in the presence the storage days. A time that the cheese is produced under excellent sanitary conditions hygienical, it is capable to reach the period of 30 days of shelf life when kept 4° C.

Thesis (Msc Food Sc (Food Science))--University of Stellenbosch, 2009.
Grape juice is transformed into wine through the complex processes of alcoholic and malolactic fermentation that is performed by yeasts, lactic acid bacteria and acetic acid bacteria. However, the microbes involved in these processes do not only take part in ensuring the successful production of wine, but also cause spoilage of the wine if their growth is not controlled. Conventional, culture-dependent methods of microbiology have been used as the main technique in detecting and identifying these spoilage microbes. Cultureindependent techniques of molecular biology have recently become more popular in detecting possible spoilage microbes present in must and wine, since it allows the detection and identification of viable, but non-culturable microbes and are not as timeconsuming as conventional microbiological methods. The aim of this study was to investigate the sustainability of polymerase chain reaction (PCR)-based denaturing gradient gel electrophoresis (DGGE) analysis in detecting wine spoilage microbes inoculated into sterile saline solution (SSS) (0.85% (m/v) NaCl) and sterile white wine and red wine as single microbial species and as part of mixed microbial populations. Three methods of DNA isolation from SSS, sterile white wine and sterile red wine inoculated with reference microbial strains were compared in terms of DNA concentration and purity, as well as simplicity of the technique. These three DNA isolation methods were the TZ-method, the proteinase K-method and the phenol extraction method. DNA could not successfully be isolated from red wine using any of the three DNA isolation methods. The TZ-method was the method of choice for the isolation of DNA from inoculated SSS and sterile white wine as this technique gave the best results in terms of simplicity, DNA concentration and purity. PCR and DGGE conditions were optimised for the universal primer pair, HDA1-GC and HDA2, the wine-bacteria specific primer pair, WBAC1-GC and WBAC2, and the yeast specific primer pair, NL1-GC and LS2. DNA from Acetobacter pasteurianus, Lactobacillus plantarum, Pediococcus pentosaceus, Oenococcus oeni, Brettanomyces bruxellensis and Saccharomyces cerevisiae were amplified with the appropriate primers and successfully resolved with DGGE analysis. PCR and DGGE detection limits were successfully determined when 106 cfu.ml-1 of the reference microbes, A. pasteurianus, Lb. plantarum, Pd. pentosaceus and B. bruxellensis were separately inoculated into SSS and sterile white wine. It was possible to detect low concentrations (101 cfu.ml-1) with PCR for A. pasteurianus, Lb. plantarum, Grape juice is transformed into wine through the complex processes of alcoholic and malolactic fermentation that is performed by yeasts, lactic acid bacteria and acetic acid bacteria. However, the microbes involved in these processes do not only take part in ensuring the successful production of wine, but also cause spoilage of the wine if their growth is not controlled. Conventional, culture-dependent methods of microbiology have been used as the main technique in detecting and identifying these spoilage microbes. Cultureindependent techniques of molecular biology have recently become more popular in detecting possible spoilage microbes present in must and wine, since it allows the detection and identification of viable, but non-culturable microbes and are not as timeconsuming as conventional microbiological methods. The aim of this study was to investigate the sustainability of polymerase chain reaction (PCR)-based denaturing gradient gel electrophoresis (DGGE) analysis in detecting wine spoilage microbes inoculated into sterile saline solution (SSS) (0.85% (m/v) NaCl) and sterile white wine and red wine as single microbial species and as part of mixed microbial populations. Three methods of DNA isolation from SSS, sterile white wine and sterile red wine inoculated with reference microbial strains were compared in terms of DNA concentration and purity, as well as simplicity of the technique. These three DNA isolation methods were the TZ-method, the proteinase K-method and the phenol extraction method. DNA could not successfully be isolated from red wine using any of the three DNA isolation methods. The TZ-method was the method of choice for the isolation of DNA from inoculated SSS and sterile white wine as this technique gave the best results in terms of simplicity, DNA concentration and purity. PCR and DGGE conditions were optimised for the universal primer pair, HDA1-GC and HDA2, the wine-bacteria specific primer pair, WBAC1-GC and WBAC2, and the yeast specific primer pair, NL1-GC and LS2. DNA from Acetobacter pasteurianus, Lactobacillus plantarum, Pediococcus pentosaceus, Oenococcus oeni, Brettanomyces bruxellensis and Saccharomyces cerevisiae were amplified with the appropriate primers and successfully resolved with DGGE analysis. PCR and DGGE detection limits were successfully determined when 106 cfu.ml-1 of the reference microbes, A. pasteurianus, Lb. plantarum, Pd. pentosaceus and B. bruxellensis were separately inoculated into SSS and sterile white wine. It was possible to detect low concentrations (101 cfu.ml-1) with PCR for A. pasteurianus, Lb. plantarum, iv Pd. pentosaceus, and B. bruxellensis in SSS when amplified with the HDA1-GC and HDA2 primer pair. A PCR detection limit of 102 cfu.ml-1 was determined in sterile white wine for Pd. pentosaceus and 103 cfu.ml-1 for B. bruxellensis using this primer pair. The results obtained from the PCR amplification with the WBAC1-GC and WBAC2 primer pair compared well with the results of the HDA1-GC and HDA2 primer pair. The results from the DGGE detection limits indicated that it was possible to detect lower concentrations (101 – 102 cfu.ml-1) of A. pasteurianus, Lb. plantarum and Pd. pentosaceus with the HDA1-GC and HDA2 primer pair than the WBAC-GC and WBAC2 primer pair (102 – 104 cfu.ml-1). Lower detection limits were also determined for B. bruxellensis amplified with the HDA1-GC and HDA2 primer pair (103 – 104 cfu.ml-1) than with the NL1-GC and LS2 primer pair (105 cfu.ml-1). PCR and DGGE detection limits for the inoculation of A. pasteurianus, Lb. plantarum and B. bruxellensis at an inoculum of 108 cfu.ml-1 as part of mixed populations in SSS and sterile white wine compared well with the results obtained from the reference microbes inoculated as single microbial species. PCR detection limits of 101 cfu.ml-1 were determined for all three reference microbes inoculated as part of mixed populations when amplified with the HDA1-GC and HDA2 and the WBAC1-GC and WBAC2 primer pairs. It was observed that similar or higher DGGE detection limits were obtained for the reference microbes inoculated in sterile white wine (101 – 107 cfu.ml-1) than when inoculated into SSS (101 – 105 cfu.ml-1). PCR-based DGGE analysis proved to be a technique that could be used successfully with the universal, wine-bacteria and yeast specific primer pairs for the detection of A. pasteurianus, Lb. plantarum, Pd. pentosaceus and B. bruxellensis. The culture-independent technique makes the early detection of possible spoilage microbes at low concentrations in wine possible.

Escherichia coli produtoras de toxina de Shiga (STEC) são consideradas importantes patógenos de origem alimentar que apresentam o trato intestinal de ruminantes domésticos, principalmente bovinos, seu reservatório natural. Esses microrganismos estão associados a doenças severas em humanos, tais como colíte hemorrágica (CH) e síndrome urêmica hemolítica (SHU). Este trabalho teve como objetivos avaliar a ocorrência de STEC em diferentes fontes, ambientais ou não, da criação e abate de bovinos confinados. Além disso, detectar a presença dos genes stx1, stx2, ehxA e eaeA; identificar cepas O157:H7 através da pesquisa do gene uidA; evidenciar a capacidade de produção de Stx e de Eh; identificar variantes de stx e de eaeA; e determinar os sorotipos e a diversidade genética das cepas de STEC. A avaliação da presença dos genes (stx1, stx2, ehxA, eaeA e uidA) e da produção de Eh foi utilizada como triagem para a seleção de cepas possivelmente patogênicas, sendo que do total de 628 isolados avaliados, foram selecionadas 50 cepas STEC e 12 consideradas como EPEC atípicas. Das STEC, 76% foram isolados provenientes de amostras de fezes, enquanto 18% foram de amostras de carcaças e 6% de amostras de água da baia. Seis cepas isoladas de fezes e 1 de carcaça foram sorotipificadas como O157:H7, todas positivas para a presença do gene uidA. Além do sorogrupo O157, nenhum outro, dentre os principais causadores de surtos e casos esporádicos de CH e SHU, foi detectado. Das 30 cepas que apresentaram resultado positivo no ensaio de citotoxicidade em células Vero, 96,7% apresentaram gene para a produção de Stx. Em 17 das STEC foi possível identificar o tipo de Stx produzida, através de ensaio imunocromatográfico, sendo que todas apresentaram os genes correspondentes à toxina identificada, com exceção de uma cepa de carcaça que foi positiva para a produção de ambas as toxinas, mas apresentando apenas o gene stx2. Através da análise por PFGE, observou-se a disseminação e permanência de cepas STEC entre os animais. Dentre as 50 cepas STEC, 28% foram positivas para a variante Stx2d ativável e das 21 cepas eaeApositivas apenas em 8 foram detectadas variantes desse gene, sendo 7 positivas para eae-γ e a outra cepa positiva para eae-β). Através dos resultados obtidos, podemos dizer que a pesquisa do gene uidA pode ser considerada uma ótima ferramenta na triagem de isolados do sorotipo O157:H7. Por outro lado, o gene ehxA e a produção de Eh não se mostraram como bons marcadores para pesquisa de cepas Stx positivas. Houve uma ampla diversidade de sqrotipos/sorogrupos entre as cepas STEC típicas. É importante salientar que, neste estudo, STEC O157:H7 foi detectada pela primeira vez no Brasil em amostra de carcaça de bovino criado em confinamento. A detecção de cepas STEC em amostras de fezes e principalmente em amostras de carcaças de bovinos demonstra um potencial risco à saúde pública, uma vez que tais cepas podem contaminar e chegar viáveis ao produto final.
Shiga toxin (Stx)-producing Escherichia coli (STEC) are considered important foodborne pathogens that have the intestinal tract of ruminants, in particular cattle, as reservoir. These microorganisms are associated with severe human diseases as hemorrhagic colitis (HC) and hemolytic-uremic syndrome (HUS). The aims of this research were to evaluate the occurrence of STEC from different sources during the feedlot cattle breeding and slaughtering; detecting the presence of stx1, stx2, ehxA and eaeA genes; identifying O157:H7 strains through uidA; evidencing Stx and Eh production capacity; identifying stx and eaeA variants and determining STEC strains serotypes and genetic diversity. The potentially pathogenic strains were screened by detection of stx1, stx2, ehxA, eaeA and uidA, and Eh production, amongst 628 isolates studied. Fifty isolates were identified as STEC and 12 others as atypical EPEC. Among the STEC isolates, 76% were from feces, 18% from carcasses and 6% from water samples. Six strains isolated from feces and one from carcass were serotyped as O157:H7, ali being positive for the uidA. No other serogroup linked to outbreaks or sporadic cases of HC and HUS were found. From the 30 strains that showed cytotoxic effect on Vero cells, the great majority (96.7%) was positive for stx. Using an immunochromatographic assay, it was possible to identify the type of Stx produced by 17 out of the 50 STEC strains. All but one of these strains harbored the gene correspondent to the identified toxin. The other strain, even though producing both toxins, presented only stx2. It was possible to determine by PFGE the dissemination and persistence of STEC strains among the animals. 14/50 (28%) STEC strains were positive for the variant Stx2d activatable. Amongst 21 eaeA-positive strains, the variants of this gene were detected only in eight, being seven positive for eae-γ and the other eae-β. The results showed that uidA gene can be considered an excellent tool for screening O157:H7 strains. On the other hand, ehxA and Eh production, could not be considered as good markers for Stx-positive strains detection. A great diversity of serotypes/serogroups was observed among typical STEC strains. It is important to notice that this is the first report of O157:H7 strains in carcasses trom feedlot cattle in Brazil. The detection of STEC strain in fecal samples and in carcasses trom feedlot cattle evidences the potential public health risk, once these strains can contaminate the final product.

O leite é um dos alimentos mais completos da natureza e sua importância é baseada em seu elevado valor nutritivo, como riqueza de proteínas, vitaminas, gordura, sais minerais e a alta digestibilidade. Esses fatores são relevantes para considerá-lo um excelente meio de cultura para a maioria dos microrganismos. A pasteurização é necessária e tem a finalidade de eliminar os microrganismos patogênicos, além de diminuir ao máximo o número de microrganismos em geral, mas alguns deles ainda podem sobreviver ao tratamento térmico aplicado. O objetivo do presente trabalho foi avaliar a condição microbiológica e a eficácia da pasteurização industrial do leite tipos A, B, C e as condições microbiológicas do leite cru, através da enumeração de bactérias aeróbias mesófilas, termófilas e psicrotróficas, Staphylococcus coagulase positiva, determinação do Número Mais Provável (NMP) de coliformes totais e fecais e pesquisa de Salmonella spp. em 30 amostras de leite comercializado no município de Piracicaba- SP. Com base na legislação do DIPOA (Brasil, 2002) os resultados mostraram que 44,4% das amostras de leite tipo A apresentaram-se fora do padrão microbiológico para aeróbios mesófilos. Para coliformes totais e fecais, o mesmo leite apresentou 66,7% e 55,6% das amostras respectivamente, em desacordo com a legislação vigente. No leite tipo B, nenhuma amostra esteve fora do padrão microbiológico em relação a aeróbios mesófilos. Já para coliformes totais e fecais, 33,3% das amostras estiveram em desacordo com a legislação em vigor. O leite tipo C foi o que apresentou resultado mais satisfatório, pois ao contrário do que se esperava, nenhuma amostra esteve fora do padrão em relação às análises microbiológicas realizadas no presente trabalho. O leite cru analisado apresentou valores elevados para todas as análises microbiológicas realizadas. Não foram encontradas amostras contaminadas com Salmonella spp. em todas as amostras de leite analisadas. O maior valor encontrado para Staphylococcus coagulase positiva foi 1,1 x 102 UFC/mL, portanto, longe da dose infectiva, mas esse fato não deixa de ser preocupante, já que o leite é um ótimo substrato para bactérias. Após a pasteurização (62,8ºC/30’) realizada no Laboratório de Microbiologia de Alimentos, do Departamento de Agroindústria, Alimentos e Nutrição, da ESALQ, todas as amostras novamente analisadas para os parâmetros microbiológicos citados, se mostraram em acordo com a legislação nacional vigente (Brasil, 2002). Os resultados encontrados na presente pesquisa podem ser indicativos de prováveis falhas do binômio tempo/temperatura durante a pasteurização industrial; matéria-prima excessivamente contaminada; higienização e sanificação deficientes das linhas de produção ou contaminação pós-pasteurização.
Milk is one of the most valuable of all foods in nature and its importance is based on its high nutritive value. Milk is rich in proteins, vitamins, fat, mineral salts, and high digestibility. These factors are relevant for it to be considered an excellent culture medium for most microorganisms. Pasteurization is necessary and its main purpose is to eliminate pathogenic microorganisms, in addition to reducing to the maximum the number of microorganisms in general. However, some of them may still outlive the thermal treatment applied. The aim of this project was to evaluate the microbiological conditions and efficiency of the industrial pasteurization of type-A,-B and-C milk, and the microbiological conditions of raw milk, through the number of mesophillic, thermophilic and psychrotrophic aerobe bacteria, Staphylococcus positive coagulase, through the determination of the Most Probable Number (MPN) of total and fecal coliforms, and also through the research on Salmonella spp. in 30 samples of milk commercialized in Piracicaba-SP. Based on DIPOA legislation (Brazil, 2002), the results showed that 44.4% of the type-A milk samples did not meet the microbiological standard for mesophillic aerobe for total and fecal coliforms, 66.7% and 55.6% of the same milk samples, respectively, were not in accordance with the present legislation. For the type-B milk, no sample failed to meet the microbiological standard in relation to mesophillic aerobes. However, for total and fecal coliforms, 33.3% of the samples were in disagreement with the present legislation. Type-C milk was the one presenting the best result. All samples were in accordance with the microbiological analysis performed in this work. The raw milk examined showed high values for all microbiological analyses. No infected samples with Salmonella spp. were found in the analyzed milk samples. The greatest value found for Staphylococcus positive coagulase was 1.1 x 102 UFC/mL, thus, far from the infective dose, although somewhat concearning since milk is considered na excellent environment for the infestation of bacteria. After the pasteurization (62.8ºC/30’) performed in the Food Microbiology Laboratory, of the Agroindustry, Foods and Nutrition Department, ESALQ/USP, all samples that analyzed again, based on the microbiological parameters mentioned above, met the national legislation in force (Brazil, 2002). The results found in the research may be an indication of probable failures regarding time/temperature during the industrial pasteurization; infected raw-material; defecient sanitation of product lines or post-pasteurization contamination.

Thesis (MSc Food Sc (Food Science))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: This study focussed on the sensory effects of the main volatile compounds produced by Brettanomyces yeast causing spoilage in wine. This research firstly aimed to determine the detection thresholds of eight Brett-related spoilage compounds in wine. The second aim was to determine the sensory effect of the four most important Brett-related compounds when present individually in wine. The third aim was to determine the sensory effects of these four compounds when present in wine in a range of combinations, and to further investigate their effect on consumer liking. Finally, this project aimed to investigate the incidence of these compounds in a small range of South African wines. The sensory detection thresholds of 4-ethylphenol, 4-ethylguaiacol, 4-ethylcatechol, 4- vinylphenol, 4-vinylguaiacol, isovaleric acid, isobutyric acid and acetic acid were determined. Apart from 4-ethylcatechol, these values generally agreed well with recent literature where values determined in wine are available. However, the discrepancies highlighted the importance of the effect of the medium (wine) when determining sensory detection thresholds. The use of the median as alternative calculation method was also investigated, and it was found that this method gives more insightful results than the standard American Society of Testing Materials (ASTM E679-04) method. Four compounds, namely 4-ethylphenol, 4-ethylguaiacol, 4-ethylcatechol and isovaleric acid were profiled individually in wine using a trained sensory panel. It was found that all four compounds caused a suppression of the natural berry-like character in the wine, which induced a sick-sweet character. 4-ethylphenol contributed Elastoplast™ and leather aromas in the wine, both of which are commonly associated with Brettanomyces taint. 4-ethylguaiacol added a medicinal aroma to the wine, and 4-ethylcatechol and isovaleric acid were responsible for savoury and pungent aromas, respectively. 4-ethylphenol, 4-ethylguaiacol, 4-ethylcatechol and isovaleric acid were also profiled in combination according to the central composite design. Several univariate and multivariate methods were applied to the dataset obtained. PARAFAC, a multiway method not widely utilized regarding sensory data, was applied to the data, the results of which were complementary to those obtained during univariate and multivariate analyses. It was found that there is a great deal of interaction between the four compounds profiled in terms of sensory effects. The most notable was the Elastoplast™ attribute, the intensity of which was affected by all four compounds. The pungent attribute was also affected by the 4-ethylphenol concentration. Consumer analysis revealed that some of the samples spiked with Brettanomyces-spoilage compounds were preferred to the unspiked (control sample). However, no further relationship could be found between consumer liking and either chemical composition or sensory profile. It is therefore speculated that consumer liking of Brettanomyces infected wine is driven by more complex sensory or socio-demographic factors. Finally, the concentration of 4-ethylphenol, 4-ethylguaiacol, 4-ethylcatechol, 4-vinylphenol, 4- vinylguaiacol, isovaleric acid, isobutyric acid and acetic acid was determined in a small set of South African wines, selected to contain a high proportion of wines spoiled by Brettanomyces. Significant correlations were found between 4-ethylphenol and 4-ethylguaiacol, as well as 4- ethylphenol and isovaleric acid. However, no correlation could be found between 4-ethylphenol and 4-ethylcatechol. It is speculated that this lack of relationship is due to the different precursor profiles present in the analysed wines. This study paved the way for future investigations on the sensory effects of Brettanomyces spoilage in Pinotage red wine.
AFRIKAANSE OPSOMMING: Hierdie studie het gefokus op die sensoriese invloed van die belangrikste vlugtige komponente wat deur die Brettanomyces gis geproduseer word en bederf veroorsaak in wyn. Eerstens is gefokus op die bepaling van die deteksiedrempelwaardes van agt Brett-verwante bederwende komponente. Die tweede doelwit was om die sensoriese invloed van vier van die mees belangrike Brett-komponente te bepaal wanneer hulle individueel in wyn voorkom. Die derde doelwit was om die sensoriese invloed van hierdie vier komponente te bepaal wanneer hulle in verskillende kombinasies in wyn voorkom, asook die effek daarvan op verbruikervoorkeur. Laastens is gepoog om die voorkoms van hierdie komponente in ‘n klein seleksie van Suid- Afrikaanse wyne te bepaal. Die sensoriese deteksiedrempelwaardes vir 4-etielfenol, 4-etielguaiacol, 4-etielcatechol, 4- vinielfenol, 4-vinielguaiacol, isovaleraatsuur, isobuteraatsuur en asynsuur is bepaal. Met die uitsondering van 4-etielcatechol het die waardes oor die algemeen goed ooreengestem met waardes wat onlangs in die wetenskaplike literatuur gepubliseer is. Die uitsonderings het egter die belangrikheid van die medium (wyn) gedurende die bepaling van sensoriese deteksiedrempelwaardes uitgelig. Die gebruik van die mediaan as ‘n alternatiewe berekeningsmetode is ook ondersoek en daar is gevind dat hierdie metode meer insiggewende resultate lewer as die standaard American Society of Testing Materials (ASTM E679-04) metode. Vier komponente naamlik 4-etielfenol, 4-etielguaiacol, 4-etielcatechol en isovaleraatsuur is individueel in wyn geprofileer met behulp van ‘n opgeleide sensoriese paneel. Daar is gevind dat al vier die komponente die natuurlike bessiekarakter in die wyn onderdruk terwyl dit aanleiding gee tot ‘n onnatuurlike soet karakter. 4-etielfenol is gekenmerk aan Elastoplast™ en leeragtige aromas in die wyn en beide van hulle word algemeen geassosieer met Brettanomyces bederf. 4-etielguaiacol het ‘n medisinale aroma tot die wyn toegevoeg en 4- etielcatechol en isovaleraatsuur het respektiewelik souterige (“savoury”) en sterk (“pungent”) aromas tot gevolg gehad. 4-etielfenol, 4-etielguaiacol, 4-etielcatechol en isovaleraatsuur is ook in verskeie kombinasies geprofileer volgens die sentrale saamgestelde ontwerp (“central composite design”). Verskeie enkelveranderlike en meerveranderlike statistiese analisemetodes is ook op die datastel uitgevoer. PARAFAC, ‘n meerrigtingsmetode wat nie normaalweg vir sensoriese analise data gebruik word nie, is ook uitgevoer op die data en die resultate was komplimentêr tot die van die enkelveranderlike en meerveranderlike analisemetodes. Daar is gevind dat, met betrekking tot sensoriese effekte, daar noemenswaardige interaksie tussen die vier komponente plaasvind. Die mees opmerklike hiervan was die Elastoplast™ aroma, waarvan die intensiteit deur al vier die ander komponente geaffekteer is. Verder is die sterk (“pungent”) aroma beïnvloed deur die 4-etielfenol konsentrasie. Verbruikersvoorkeur-analise het aangedui dat sommige van die monsters waarby Brettanomyces bederwende komponente gevoeg is, verkies word bó die kontrole-wyn. Daar kon egter geen verdere verband gevind word tussen die verbruiker se voorkeur en, nog die chemise komposisie of sensoriese profiele, van die wyn nie. Daar kan dus gespekuleer word dat verbruiker voorkeur van Brettanomyces bederfde wyn gedryf word deur meer komplekse en sosio-demografiese faktore. Laastens is die konsentrasies van 4-etielfenol, 4-etielguaiacol, 4-etielcatechol, 4-vinielfenol, 4- vinielguaiacol, isovaleraatsuur, isobuteraatsuur en asynsuur in ‘n seleksie van Suid-Afrikaanse wyne bepaal. Dié wyne is spesifiek so gekies sodat ‘n aansienlike aantal van hulle met Brettanomyces bederf was. Betekenisvolle korrelasies is gevind tussen 4-etielfenol and 4- etielguaiacol, sowel as 4-etielfenol en isovaleraatsuur. Daar is egter geen korrelasie tussen 4- etielfenol and 4-etielcatechol gevind nie. Daar word vermoed dat hierdie gebrek aan korrelasie te wyte is aan die voorloperkomponent profiele teenwoordig in die wyne. Hierdie studie het die weg gebaan vir verdere ondersoeke na die sensoriese effekte van Brettanomyces bederf in Pinotage rooi wyn.

Nesta pesquisa, estudaram-se as variações ocorridas na relação entre autólise de culturas lácticas e o desenvolvimento da proteólise de queijo Prato produzido em quatro regiões brasileiras: Santa Catarina (Queijo A), Goiás (Queijo B), São Paulo (Queijo C) e Minas Gerais (Queijo D). A análise quantitativa da população de bactérias lácticas durante a maturação mostrou perfis microbiológicos similares para todas as amostras de queijos examinadas. Após 5 dias de maturação, lactococos e estreptococos estavam presentes em números mais elevados do que lactobacilos mesofílicos e termofílicos, leuconostoc e fermentadores de lactato. Contudo, essas populações aumentaram consideravelmente no final do processo de maturação. Enterococos e fermentadores de citrato permaneceram em números relativamente reduzidos ao longo da maturação. A análise qualitativa mostrou a predominância de \"non starter lactic acid bactéria\" (NSLAB) nos queijos das quatro origens, principalmente de Lactobacillus sp. Outros gêneros foram identificados em menor proporção: Enterococcus sp., Pediococcus sp., Aerococcus sp., Tetragenococcus sp. e Streptococcus sp. O queijo C diferiu dos demais por não apresentar Pediococcus sp. e Streptococcus sp. As culturas lácticas adicionadas Lactococcus lacfis sp. e Leuconostoc sp. estavam presentes em populações menores. A autólise foi estudada pela determinação da atividade de aminopeptidases e detecção de autolisinas por zimogramas e de enzimas intracelulares por \"imunoblotting\". Uma maior liberação de aminopeptidases ocorreu no queijo D, seguido dos queijos C, B e A. Não foram detectadas bandas de atividade lítica nos zimogramas dos queijos A e B em todas as condições avaliadas. Nos zimogramas, detectou-se uma banda de 30 KDa nos queijos C e D a pH 7,4 e 44°C, e uma outra de 40 KDa, exclusiva no queijo D, ambas de fraca intensidade. A pH 6,8 e 42°C, detectou-se bandas de 40KDa de fraca intensidade no queijo C e forte no D, além de mais duas de fraca intensidade de 90KDa e 110KDa no queijo D. Na análise em \"imunnoblotting\" com o antisoro anti-Lc, foi observado apenas sinais fracos de reação positiva e em números inferiores àquelas reveladas com o citoplasma bruto de Lac. Lacfis subsp. lacfis (controle positivo), indicando que a autólise foi praticamente inexistente. Com o antisoro anti-D-LDH, também não se detectou sinais de reação positiva nos queijos A e B, enquanto nos queijos C e D, verificou-se sinais positivos de 37KDa, de forte intensidade e correspondentes à proteína D-LDH, indicando a lise de Lab. helveticus. A evolução da proteólise foi determinada quantitativamente durante a maturação e avaliada com base nos índices: NS-pH4,6/NT% e NNP/NT%, teor de tirosina, eletroforese (Uréia-PAGE) e quantificação de aminoácidos individuais livres. Não foram detectadas diferenças significativas entre os queijos A. B, C e D no início da maturação. Contudo, com a fragmentação das proteínas, ocorreu um aumento gradual desses índices, tendo-se observado valores mais elevados no queijo D, seguido dos queijos C, B e A. Os perfis eletroforéticos de proteínas foram similares para os queijos das quatro origens e mostraram claramente que o coagulante e a plasmina foram os responsáveis pela degradação inicial das caseínas. A taxa de degradação da αs1- e β-caseína apresentou a seguinte ordem: D > C ≥ B > A. O acúmulo de aminoácidos livres também foi mais rápido no queijo D, seguido dos queijos C, B e A. Portanto, a autólise de Lab. helveticus no queijo D acelerou a proteólise, diminuindo o período de maturação em 45% e não afetando negativamente o desenvolvimento de \"flavour\" e nem a textura. No final da maturação (45 dias), os compostos voláteis foram determinados por meio de cromatografia gasosa com espectrometria de massa (CG-MS). Com raras exceções, os queijos das quatro origens continham os mesmos compostos voláteis, embora em quantidades distintas. Os álcoois e ésters foram os compostos majoritários nos queijos A e B e benzaldeído, 3-metil-butanal-2 e hexanal nos C e D. O perfil de textura instrumental (TPA) e a análise sensorial descritiva e quantitativa foram realizados. Os queijos B, C e D apresentaram características mais típicas de queijo Prato, independentemente do fato de que o aroma de manteiga e o sabor doce serem mais acentuados no queijo D. O queijo A foi classificado como tendo as menores características de queijo Prato e apresentou maior nível de defeitos de \"flavour\", principalmente residual e amargor. Os queijos avaliados não apresentaram diferenças significativas quanto à elasticidade e coesividade. Pequenas alterações na composição físico-química dos queijos, principalmente os teores de umidade e de caseína, influenciaram nos parâmetros como a firmeza e a adesividade. O presente estudo demonstrou pela primeira vez a ausência de autólise de Lc. Lacfis sp. em queijo Prato de quatro origens, bem como a ocorrência de autólise de Lab. helveticus nos queijos de duas origens, C e D. A pronunciada autólise dessa espécie teve um impacto positivo na proteólise e foi a responsável pelo aumento da concentração de aminoácidos livres nesses queijos. As diferenças na evolução da proteólise observada entre os queijos C e D, com taxas mais baixas no queijo C, independentemente da autólise pronunciada de Lab. helveticus, foram atribuídas à falta de uniformidade na composição físico-química dos queijos, principalmente pH e os teores de sal na umidade (S/U).
This paper reports a study aimed at evaluating the variations that occur in the interrelationship between autolysis of lactic starter bacteria and the development of proteolysis in Prato cheese produced in four different regions of Brazil: Santa Catarina (Cheese A), Goiás (Cheese B), São Paulo (Cheese C) and Minas Gerais (Cheese D). Quantitative analysis of microbial population yielded similar microbiological profiles for all the cheese samples investigated. After 5 days ripening, lactococci and streptococci were present in higher numbers than mesophilic and thermophilic lactobacilli, leuconostoc and lactate fermenting bacteria. However, the populations of the latter species had considerably increased by the time the ripening process completed 45 days. Enterococci and citrate fermenting bacteria remained present in relatively low numbers throughout ripening. The findings from qualitative analysis confirmed the predominance of non-starter lactic acid bacteria (NSLAB) in the cheeses from four different origins, especially Lactobacillus sp. Other genera of non-starter lactic acid bacteria (NSLAB) were identified in smaller proportions: Enterococcus sp., Pediococcus sp., Aerococcus sp., Tetragenococcus sp. and Streptococcus sp. Cheese C differed from the cheeses in that it no evidence was found of the presence of Pediococcus sp. and Streptococcus sp. The bacteria of the lactic starter culture Lactococcus lactis sp. and Leuconostoc sp. were also found to be present, although in lower numbers. Autolysis was studied by: (1) determination of aminopeptidase activity; (2) detection of autolysins by zymograms and (3) detection of intracellular enzymes by immunoblotting. The release of aminopeptidase was highest in cheese D, followed by C, B and A. No bands of lytic activity were appeared in the zymograms of Cheeses A and B in all conditions evaluated. At pH 7,4 and 44°C, a low-intensity band of 30 KDa was found in cheeses C e D, whereas another low-intensity band was observed only in cheese D. At pH 6,8 and 42°C, bands of 40KDa were observed in cheese C (low intensity) and cheese D (high intensity), in addition to two more low-intensity bands of 90KDa and 110KDa in cheese D. Immunoblotting with antiserum anti-Lc produced only minor signs of positive reaction, evidenced by the formation of low-intensity bands of 100 Kda in cheeses A and B and two high-intensity bands of 75 Kda and 100 Kda in cheeses C and D. Since these were present in smaller numbers to those revealed with crude cytoplasm of Lac. lactis subsp. lactis, it was concluded that autolysis did practically non occur. Immunoblotting with antiserum anti-D-LDH also detected sings of positive reaction in cheeses A and B, whereas in cheeses C e D positive high-intensity signs of 37Kda were found relative to D-LDH protein, indicating lysis of Lab. helveticus. The evolution of proteolysis was determined quantitatively during the ripening process and evaluated on the basis of the following parameters: NS-pH4,6/NT% and NNP/NT% indexes, tyrosine content, electrophoresis (Urea-PAGE) and quantification of free amino acids. No significant differences were found between cheeses A, B, C and D in the ear1y stages of ripening. However, with the on-going fragmentation of proteins during ripening, a gradual increase of the ripening indexes occurred, with the highest values being observed in cheese D, followed by C, B e A. The electrophoretic profiles were similar for the four cheeses investigated and clear1y showed that the clotting agent or milk coagulant and plasmin were responsible for the initial breakdown of the caseins. The degradation rate of Q.sl- and p-casein followed the following order: D > C ≥ B > A. The buildup of free amino acids was also faster in cheese D, followed by cheeses C, B e A. At the end of the ripening process studied (45 days), the volatile compounds were identified using gas chromatography and mass spectrometry (GC-MS), whereas the instrumental texture profile was measured and evaluated by Texture Profile Analysis (TPA). Cheese samples were evaluated by descriptive and quantitative sensory analysis. With rare exceptions, the cheeses of four different origins contained the same volatile compounds, although in different quantities. Alcohols and esters were the predominant volatile compounds in cheeses A and B and benzaldehyde, 3-methyl-butanal-2 and hexanal in cheeses C and D. Autolysis of Lb. helveticus accelerated proteolysis in cheese D, thereby reducing ripening time by 45% without any negative effect on either flavor or texture development. Cheeses B, C and D exhibited the most typical Prato cheese characteristics, in spite of the fact that the buttery aroma and sweet taste were more pronounced in cheese D. Cheese A was rated as the cheese with the less typical overall Prato cheese profile and was also the one that exhibited the highest degree and number of flavor defects, notably aftertaste and bitterness. The cheeses investigated did not present any significant differences as to elasticity and cohesiveness. Minor changes in the physical-chemical composition of the cheeses - mainly related to the moisture and casein levels - influenced parameters such as firmness and adhesiveness. The present study demonstrates for the very first time the absence of autolysis of Lc. Lactis sp. in Prato cheese from four different origins, as well as the occurrence of autolysis of Lb. helveticus in two of the cheeses analyzed (cheeses C and D). The pronounced autolysis of this species had a positive impact on proteolysis and was responsible for the release of increased quantities of free amino acids in these cheeses. The differences in the evolution of proteolysis observed between cheeses C and D - lower rate of proteolysis in cheese C, in spite of pronounced autolysis of Lb. helveticus - were attributed to poor uniformity of the physical-chemical composition of this cheese, particularly as related to pH and the salt and moisture levels (S/M).

The poultry industry is seeking an advanced chilling system that can improve chilling efficiency, microbial safety, and water consumption without compromising meat quality. The objective of this study was to evaluate the effects of sub-zero saline chilling methods on chilling efficiency, breast fillet tenderness and microbial reduction of broiler carcasses. Following evisceration and rinsing, broiler carcasses were randomly assigned to one of three chilling solutions: 1) 0% salt or ice water control (0% NaCl/0.5oC), 2) 3% salt (3% NaCl/-1.8oC), and 3) 4% salt (4% NaCl/-2.41oC) solutions. Broiler carcasses in sub-zero saline solutions reached the target internal temperature of < 4.4 oC in a faster rate than the 0% salt control, reducing the chilling time by 11% and 39 % for 3% NaCl/-1.8oC and 4% NaCl/-2.41oC solutions, respectively. There was no significant difference in breast fillet pH, regardless of chilling treatment (P < 0.05). However, the breast fillets from sub-zero saline solutions showed higher R-value and longer sarcomere length than those of control fillets (P < 0.05). Breast fillets excised from carcasses in 4% NaCl/2.41oC were significantly tenderized more than the control fillets, with an intermediate tenderness observed for the fillets from 3% NaCl/-1.8oC (P< 0.05). Before chilling, broiler carcasses contained mesophilic aerobic bacteria (MAB), Escherichia coli(E. coli), and total coliforms for 3.81, 0.78, and 1.86 log colony forming unit (CFU)/g, respectively. After chilling, the populations of E. coliand total coliforms were significantly reduced on the carcasses in 3% NaCl/-1.8oC and 4% NaCl/-2.41oCcompared to the control fillets (P< 0.05). There was no significant difference for MAB populations, regardless of treatment. Based on these results, chilling of broiler carcasses in 4% NaCl/-2.4 °C solution seems to be the best choice to improve chilling efficiency, meat tenderness, and microbial reduction compared to the control (0% NaCl/0.5ºC) and 3% NaCl/-1.8oCsolutions.

Dissertação de Mestrado Integrado em Medicina Veterinária
A transmissão de agentes patogénicos através do consumo de alimentos pode constituir um problema sério de Saúde Pública e, por isso, as questões relacionadas com a segurança alimentar têm suscitado uma crescente preocupação nas autoridades, indústrias, produtores, fornecedores e consumidores. Sinal desta preocupação é toda uma nova legislação alimentar que tem sido desenvolvida. A Análise de Risco é uma ferramenta fundamental para a implementação de sistemas de autocontrolo da cadeia alimentar, como o Sistema HACCP que, assente em sete princípios, de carácter pró-activo, sistemático e científico, valoriza a prevenção em detrimento dos testes ao produto final e tem por objectivo identificar os perigos e concentrar o controlo da produção de alimentos para consumo nos chamados pontos críticos de controlo (PCC). Igualmente indispensável, para a eficácia do Sistema HACCP, é a implementação de programas de pré-requisitos, como as boas práticas de higiene e de fabrico, assim como a formação e sensibilização de todos os operadores da cadeia alimentar para esta problemática. Tendo como exemplo um catering de aviação, viu-se realçada precisamente a importância da sólida implementação dos programas de pré-requisitos, do total envolvimento e sentido de responsabilidade dos manipuladores e da actuação dirigida, persistente e determinada da equipa HACCP responsável.
ABSTRACT - Food-borne illness is a serious Public Health problem. This fact has increased public awareness and is today a major concern to authorities, industry, production, supplying sectors and consumers, as a new food hygiene legislation is being created. Risk Analysis is a fundamental tool for the implementation of self-control systems in the food chain, such as the HACCP which is based in seven principles, has an active, systematic and scientific character prioritizing prevention instead of testing of the final product. The objective of this method is to identify hazards and center food production control in critical control points (CCP). The implementation of prerequisites, such as good manufacturing practice and good hygiene practice, are vital for the efficiency of the HACCP. The consciousness of the food industry operators, along with continuous training programs of the operators play an important role to solve the problem. This essay applies the above concepts to a flight catering business where the establishment of a self-control system, along with employee training, and a firm hold, guidance and persistent actions of the HACCP team is enhanced.

Este estudo visou analisar a qualidade microbiológica do leite humano cru recebido em um banco de leite humano (BLH), quantificando os microrganismos aeróbios mesófilos, Staphylococcus coagulase positivos (ECP), coliformes totais e Escherichia coli. Foram realizadas análises microbiológicas em 60 amostras de leite humano cru recebidas no BLH e os resultados foram os seguintes: o microrganismo E. coli foi positivo em 50% das amostras analisadas, os coliformes totais em 75% delas, o ECP em 46,66% e em 96,66% do total das amostras foram detectados microrganismos aeróbios mesófilos. Das amostras analisadas, a população de E. coli foi positiva em 93,33% (28 amostras) com uma variação entre 1 e 99 NMP/mL; 3,33% (1 amostra) entre 104 e 9,9 x 104 NMP/mL; 3,33% (1 amostra) entre 105 e 9,9 x105 NMP/mL. A população de coliformes totais que foi positiva em 57,77% (26 amostras) das amostras analisadas ficou entre 1 e 99 NMP/mL; 24,4% (11 amostras) ficaram entre 102 e 9,9x102 NMP/mL; 2,22% (1 amostra) entre 103 e 9,9x103 NMP/mL; 8,88% (4 amostras) entre 104 e 9,9x104 NMP/mL; 4,44% (2 amostras) entre 105 e 9,9x105 NMP/mL; 2,22% (1 amostra) entre 106 e 9,9x106 NMP/mL. Os microrganismos aeróbios mesófilos apresentaram-se na quase totalidade das amostras, sendo que 17,24% (10 amostras) variaram de 1 a 99 UFC/mL; 17,24% (10 amostras) entre 102 e 9,9x102 UFC/mL; 10,34% (6 amostras) ficaram entre 103 e 9,9 x 103 UFC/mL; 25,86% (15 amostras) entre 104 e 9,9 x104 UFC/mL; 17,24% (10 amostras) entre 105 e 9,9 x 105 UFC/mL; 10,34% (6 amostras) entre 106 e 9,9x106 UFC/mL e 1,72% (1 amostra) entre 107 e 9,9x107 UFC/mL. Porém, 3,33% (2 amostras) apresentaram-se negativas. Do total de amostras analisadas, o grupo dos microrganismos ECP apresentouse em 78,57% (22 amostras) com variação entre 1 e 99 UFC/mL; em 3,57% (1 amostra) entre 102 e 9,9x102 UFC/mL; 10,71% (3 amostras) entre 103 e 9,9x103 UFC/mL e 7,14% (2 amostras) entre 104 e 9,9 x 104 UFC/mL.
This paper aimed to analyze the microbiological quality of the raw human milk which is received in a human milk bank (HMB) by quantifying the aerobic mesophiles, Staphylococcus aureus (SCP), total coliforms and Escherichia coli. Sixty (60) samples of raw human milk received in HMB were microbiologic analyzed and the results were as follows: the E. coli microorganism was positive in 50% of the analyzed samples, the total coliforms were in 75% of them, the ECP in 46,6% and in 96,6% of the total amount of samples aerobic mesophiles microorganisms were detected. From the total analyzed samples, the population of E. coli was positive in 93,33% (28 samples) with a variation between 1 and 99 NMP/mL; 3,33% (1 sample) was between 104 and 9,9x104 NMP/mL; 3,33% (1 sample) was from 105 to 9,9x105 NMP/mL. The population of the total coliforms which was positive in 57,77% (26 samples) of the total analyzed samples were between 1 and 99 NMP/mL; 24,4% (11 samples) were between 102 and 9,9x102 NMP/mL; 2,22% (1 sample) were between 103 and 9,9x103 NMP/mL; 8,88% (4 samples) were between 104 and 9,9x104 NMP/mL; 4,44% (2 samples) between 105 and 9,9x105 NMP/mL and in 2,22% (1 sample) from 106 to 9,9x106 NMP/mL. The aerobic mesophiles microorganisms were presented in almost all the samples, being: 17,24% (10 samples) with a variation of 1 to 99 UFC/mL; 17,24% (10 samples) between 102 and 9,9x102 UFC/mL; 10,34% (6 samples) were between 103 and 9,9x103 UFC/mL; 25,86% (15 samples) between 104 and 9,9x104 UFC/mL; 17,24% (10 samples) between 105 and 9,9x105 UFC/mL; 10,34% (6 samples) between 106 and 9,9x106 UFC/mL and in 1,72% (1 sample) between 107 and 9,9x107 UFC/mL. However 3,33% (2 samples) showed to be negative. From the total analyzed samples, the SCP group presented in 78,57% (22 samples) with a variation between 1 and 99 UFC/mL; 3,57% (1 sample) between 102 and 9,9 x 102 UFC/mL; 10,71% (3 samples) between 103 e 9,9x103 UFC/mL and 7,14% (2 samples) were between 104 and 9,9x104 UFC/mL.

O experimento foi dividido em duas partes, sendo que inicialmente estudou-se os queijos Minas Frescal embalados sob ar atmosférico, atmosfera modificada de 70% CO2 e 30%N2 (ATM) e Vácuo e na segunda parte os queijos embalados sob os três tratamentos foram irradiados com doses de 2 KGy, sendo analisadas, nas duas partes do experimento, a evolução microbiana e as características sensoriais e físico-químicas do queijo sob os diferentes tratamentos durante o armazenamento a 4ºC. Na primeira fase do experimento verificou-se que a ATM e o Vácuo diminuíam a intensidade de crescimento da população de microrganismos mesófilos e psicrotróficos totais e reduziram a população de Staphylococcus coagulase positiva, mas não foram eficientes para o controle das populações coliformes totais e Escherichia coli, enquanto que no controle todas as populações cresceram continuamente, segundo os aspectos sensoriais de cor, odor e aparência dos queijos, mantiveram essas características durante os 40 dias de armazenamento e o controle diminuiu os índices de aceitabilidade progressivamente sendo rejeitado no 17º dia. Na segunda parte do experimento observou-se que irradiação no queijo Minas frescal com 2 KGy reduziu as populações de bactérias mesófilas aeróbias, psicrotróficas aeróbias e anaeróbias, Staphylococcus coagulase positiva, coliformes totais e Escherichia coli e os tratamentos ATM e Vácuo foram muitos eficientes pois eles evitaram o crescimento destes microrganismos durante o armazenamento, enquanto que no controle a população de mesófilos e psicrotróficos aeróbios cresceu durante o armazenamento. Segundo as características sensoriais, o tratamento ATM foi o mais eficiente pois este manteve a aparência, textura e sabor por mais de 43 dias, o vácuo por 36 dias e o controle por apenas 8 dias. A utilização da irradiação com atmosfera modificada juntamente com baixas temperaturas de armazenamento aumentaram a vida útil do queijo, impedindo o desenvolvimento microbiano no produto e mantendo suas características sensoriais.
The experiment was divided into two parts. Initially, it was studied the “Minas Frescal” cheeses packed under atmospheric air, modified atmosphere of 70% CO2 and 30% N2 (ATM) and vacuum. Second the cheeses packed under these three treatments had been radiated by doses of 2 KGy. In the two parts of the experiment, it was analyzed the microbial evolution and, the sensory and physical-chemical characteristics of the cheeses under the different treatments during a 4ºC-storage. In the first phase of the experiment it was verified that the ATM and the vacuum decreased the intensity of the total population growth of aerobic mesophilic and psichorotrophs and had reduced the population of Staphylococcus positive coagulase, but they had not been efficient controlling the total coliforms and Escherichia coli, while in control all the populations had continuously grown, according to the sensory characteristics of the cheeses, color, odor and appearance. These characteristics were kept the same during the 40 days of storage, and the control decreased the acceptability levels gradually, being rejected in the 17th day. In the second part of the experiment, it was observed that a 2KGy-irradiation over the “Minas Frescal” cheeses reduced the populations of aerobic mesophilic, aerobic and anaerobic psichorotrophs, Staphylococcus positive coagulase, total coliforms and Escherichia coli. The ATM and vacuum treatments were very efficient therefore they prevented the growth of these microrganisms during the storage, while in control, the aerobic mesophilic and psichrotrophs population grew during the storage. According to sensory aspects, the ATM treatment was the most efficient one, because it kept the appearance, texture and flavor for more than 43 days while the vacuum kept for 36 days and the control for only 8 days. The use of the irradiation with modified atmosphere and low temperatures of storage increased the shelflife of the cheeses, hindering the growth of the microbial populations in the product and keeping product sensory characteristics.

As mudanças nos hábitos de consumo e a presença de compostos com propriedades antioxidantes, capazes de reduzir o risco de doenças degenerativas, aumentaram a procura por vegetais minimamente processados. Uma vez que as doenças transmitidas por esses vegetais são um problema crescente no cenário internacional, este estudo foi conduzido com o objetivo de avaliar a viabilidade da utilização do processo de irradiação associado à embalagem sob atmosfera modificada (15% O2, 5% e O2 e 80% N2 ) em rúcula (Eruca sativa Mill.) minimamente processada para garantir a inocuidade do alimento. Nesta pesquisa constatou-se que a sanificação com ozônio (0,08 ppm/5 minutos) reduziu as populações de psicrotróficos e mesófilos aeróbios, bactérias láticas, Pseudomonas e coliformes termotolerantes em até 1 ciclo logarítimico. Não foi verificada a presença de Listeria monocytogenes ou Salmonella nas amostras analisadas, tanto antes quanto após o processamento mínimo. Os valores de D10 determinados neste estudo, para Salmonella e L. monocytogenes inoculadas em amostras de rúcula variaram de 0,16 a 0,22 kGy e de 0,33 a 0,48 kGy, respectivamente, não diferindo estatisticamente (P>0,05) em relação ao tipo de embalagem utilizada (ar atmosférico e atmosfera modificada). A combinação da aplicação de 2 kGy associada à atmosfera modificada foi o tratamento mais eficiente para reduzir a população de L. monocytogenes a níveis não detectáveis ao longo de todo o período de armazenamento (5°C por 16 dias). Tanto as amostras irradiadas quanto as controle apresentaram redução no teor de vitamina e ao longo dos 16 dias de armazenamento. Por outro lado, verificou-se aumento do teor de f1avonóides para as amostras irradiadas. Não se observou alteração no teor de carotenóides com atividade prá-vitamina A após irradiação ou período de estocagem. A análise sensorial demonstrou que a aplicação de 2 kGy não afetou a aceitação da rúcula. Além disso, a percepção da população sobre o risco da irradiação de alimentos é significativamente influenciada (P≤0,05) em função do tipo de informação adquirida. O processo de irradiação associado às Boas Práticas de Fabricação garantem a segurança microbiológica desse produto, entretanto é necessário maior divulgação sobre esta tecnologia para que ela possa ser aceita comercialmente no Brasil.
Changes in consumption habits and the presence of bioactive compounds with antioxidant capacity, which have the property to protect against degenerative disease, has increased the demand for minimally processed vegetables. Since foodbome diseases associated with these vegetables are increasing problems on the intemational scene, this study was conducted to evaluate the feasibility of associating irradiation with modified atmosphere packaging (15% O2, 5% CO2 and 80 % N2) in minimally processed arugula (Eruca sativa Mill.), to ensure the food safety. Sanitization in ozone-treated water (0.08 ppm/5 minites) reduced psychrotrophic, mesophilic, lactic acid bacteria, Pseudomonas and fecal coliform by 1 log. Listeria monocytogenes and Salmonella were not isolated in samples. D10 values for Salmonella and L. monocytogenes inoculated in arugula samples ranged frem 0.16 to 0.22 kGy and from 0.33 to 0.48 kGy, respectively, not statistically different (P> 0.05) with the type of packaging used (air and modified atmosphere). The 2 kGy dose with modified atmosphere packaging was the most efficient treatment reducing the population of L monocytogenes at non detectable levels during the storage period up to 16 days at 5°C. The vitamin C content decreased in irradiated (1 and 2 kGy) and non-irradiated samples during the storage period, on the other hand, irradiation caused a very significant increase in flavonoid content. No significant change in carotenoids with pro-vitaminic A activity content was observed after irradiation or storage period. The sensory evaluation showed that the exposition to 2 kGy did not affect the acceptance of arugula. What is more, the population risk perception of food irradiation is significantly influenced (P≤:0.05) by the type of the given information. The combination of irradiation and Good Manufacture Practices improve the microbiological safety of these products, however it is necessary to provide more information about this technology so it can be commercially acceptable in Brazil.

Os mexilhões cultivados no litoral Norte de São Paulo, município de Ubatuba, são comercializados in natura, constituindo risco à população. Com o crescimento da atividade é possível sugerir a implantação de uma unidade de processamento de mexilhões que promova um aumento do tempo de armazenamento, facilitando a comercialização e permitindo a exportação, além de fornecer ao consumidor um produto de melhor qualidade. Os mexilhões foram submetidos ao processamento por cocção, congelamento e armazenamento, sendo então determinados o ponto de congelamento, a velocidade de congelamento a as curvas de congelamento do mexilhão semidesconchado. A qualidade microbiológica e físico-química do produto foi avaliada. O beneficiamento do mexilhão iniciou-se com a cocção por imersão em água à ebulição por 10 minutos. Após a retirada das conchas, os mexilhões foram congelados individualmente IQF (Individually Quick Frozen) a –20ºC e armazenados a -18ºC durante 90 dias. A curva de congelamento do mexilhão apresentou forma geral típica, com o ponto de congelamento situando-se na faixa de zero a –1,5ºC; a velocidade de congelamento variou de 2 cm/h a 3,3 cm/h, conforme a disposição dentro da câmara de congelamento. Os resultados físico-químicos mostraram que não houve diferença significativa no valor nutricional dos mexilhões in natura, processados e armazenados, apresentando os teores médios de 7,4mg/100g de proteínas, 5,8 mg/100g de carboidratos e 1,4 mg/100g de lipídeos. Os valores encontrados para BNVT, TMA e pH no mexilhão in natura foram, 4,3 mg/100g; 2,0 mg/100g e 6,2, respectivamente, estando dentro dos limites estipulados pela legislação de 30mg/100g para BNVT e 4 mg/100g de TMA. Após o processo de cocção e congelamento houve um aumento no valor do pH para 6,9, enquanto o BNVT manteve-se na média. Todas as amostras de mexilhão in natura atenderam aos padrões microbiológicos estabelecidos pela legislação (RDC, nº12 de 02 de janeiro de 2001). Salmonella sp e Vibrio parahaemolyticus não foram isoladas em nenhuma das amostras de mexilhões in natura, cozidos, congelados e armazenados. O tratamento térmico foi efetivo no controle dos coliformes fecais, redução de coliformes totais, Staphylococcus coagulase+ e psicrotróficos. O processo de congelamento reduziu a contagem de coliformes totais e Staphyloocccus coagulase+, mantendo-se inalterados durante o armazenamento a –18ºC por 90 dias. Concluiu-se que o beneficiamento do mexilhão pelo processo combinado de cocção, congelamento e armazenamento assegura a qualidade físico-química e microbiológica do produto, podendo ser adotado como padrão para industrialização.
The mussels cultivated in the coast North of São Paulo, city of Ubatuba, are commercialized in natura, constituting risk to the population. With the growth of the activity it is possible to suggest the implantation of a unit of mussel processing that promotes an increase of the storage time, facilitating the commercialization and allowing the exportation, besides supplying to the consumer a product of better quality. The mussels had been submitted to the processing for cooking, freezing and storage, being then determined the freezing point, the speed of freezing to the curves of freezing of the semidesconchado mussel. The microbiological quality and physicochemical of the product were evaluated. The processing of the mussel was started with the immersion in boiling water per 10 minutes. After the withdrawal of the shells, the mussels had been frozen individually IQF (Individually Quick Frozen) at -20ºC and stored at -18ºC during 90 days. The curve of freezing of the mussel presented typical general form, with the freezing point placing from zero to -1,5ºC; the freezing speed varied from 2 cm.h-1 to 3,3 cm.h-1, as the disposal

Os mexilhões são alimentos marinhos freqüentemente ingeridos crus, ou parcialmente cozidos, e o hábito de aferventar estes bivalves somente até que abram as valvas, é insuficiente para eliminar os microrganismos patogênicos eventualmente presentes neste molusco. Após levantamento inicial, e visando melhorar a qualidade do mexilhão Perna perna cultivado e comercializado no município de Ubatuba, SP, esta pesquisa estudou o crescimento de Staphylococcus aureus e Bacillus cereus em mexilhões in natura e pré-cozidos, e a eliminação dos mesmos por meio de tratamentos térmicos, avaliando, posteriormente, as características físico-químicas e sensoriais dos produtos. Em ambos os casos, lotes de 1 kg de mexilhão foram inoculados, individualmente, com cepas de S. aureus e B. cereus e mantidos, por 10 horas, a temperatura ambiente (25ºC±1ºC) e sob refrigeração (7ºC±1ºC). Posteriormente, foram estabelecidos seis tipos de tratamentos térmicos, sendo três sob vapor (5, 10 e 15 min) e três por imersão em água (5, 10 e 15 min), buscando estabelecer o binômio que proporcionasse a eliminação dos mesmos, e avaliando o rendimento, os aspectos físico-químicos e sensoriais. Para ambos microrganismos, ocorreu crescimento durante as 10 horas de estudo, sendo este mais evidente, nos tratamentos mantidos a temperatura ambiente. No mexilhão pré-cozido ocorreram as maiores contagens microbianas, se comparado ao mexilhão in natura. Com relação aos tratamentos térmicos, todos foram eficientes, eliminando os microrganismos da ordem de, pelo menos 2 ciclos logarítmicos, no entanto, os tratamentos térmicos por imersão em água, permitiram melhores resultados do que os tratamentos sob vapor. As análises fisico-químicas e sensoriais, não apresentaram diferença estatística entre os tratamentos térmicos estudados. Com o emprego de altas temperaturas por um determinado período, obteve-se perda de alguns minerais, como Potássio e Boro, tendo outros, não apresentado alteração com relação ao tempo de exposição ao calor. Já, quanto ao rendimento, houve diferença, em nível de 5%, sendo os melhores rendimentos alcançados nos menores tempos de exposição ao calor e, os tratamentos por imersão, apresentaram resultados melhores que os tratamentos sob vapor. Concluiu-se que o tratamento térmico, binômio tempo-temperatura, de 10 min em água à ebulição, é suficiente para reduzir os microrganismos, permitindo a retenção dos nutrientes e um rendimento de 54,36%, podendo, portanto, ser recomendado para os produtores, visando melhorar a qualidade do mexilhão, via adequação do manejo atualmente empregado.
Mussels are seafood frequently ingested raw or partially cooked and the habit of boiling bivalves only to open the valves, is insufficient to eliminate several species of pathogenic bacteria. Seeking to improve the quality of the cultivated and marketed Perna perna mussel in the district of Ubatuba, SP, this research studied the microbiological growth of Staphylococcus aureus and Bacillus cereus in fresh and pre-cooked mussels, and the elimination by thermal treatments, being evaluated its physicochemical and sensorial characteristics. For such, lots of 1 kg of mussel was inoculated individually with strains of S. aureus and B. cereus, and maintained by 10 hours at environmental temperature (25ºC±1ºC) and under refrigeration (7ºC±1ºC). Six thermal treatments were established, 3 in steam (5, 10 and 15 min) and 3 in boiling water (5, 10 and 15 min), being looked in the elimination of B. cereus and S. aureus, and also evaluating the performance, and physical-chemical and sensorial aspects. Microbiological growth was verified after 10 hours for both microorganisms, and this being more evident in the treatments maintained at environmental temperature. Pre-cooked mussel obtained the largest microbial developments, if compared to fresh mussels. About the thermal treatments, everyone was efficient, eliminating at least 2 logarithmic cycles, however, thermal treatments in boiling water obtained better results than the steam treatments. The physical-chemical and sensorial analyses, didn't present statistical difference among the thermal treatments studied. Use of high temperatures for a determinate period, obtained lost in some minerals, like potassium and boron, and others minerals not presented alteration in relation to the heat time exposure. Already in the performance, it was obtained statistical difference, being the best performances reached in the smallest times of heat exposition, and the treatments in boiling water presented better results than the steam treatments. The thermal treatment, binomial time-temperature, of 10 min in boiling water, is enough to reduce the microorganisms, allowing the retention of the nutrients and performance of 54.36%, could be recommended for the producers seeking to improve the traditional handling.

Atualmente, as perdas econômicas são uma realidade devido ao problema de off flavor em pescado, ocasionados pela qualidade da água dos criatórios e pelo manejo empregado no cultivo, nem sempre satisfatórios. Buscou-se a avaliação microbiológica, físico-química e sensorial dos peixes, com a finalidade de detectar prováveis alterações que estivessem comprometendo a qualidade do pescado. Com o objetivo de detectar e controlar o off flavor em tilápias cultivadas foi sugerido a depuração antes da distribuição e venda e a utilização da defumação como meio de mascarar este problema. O monitoramento da qualidade do pescado iniciou-se com as análises microbiológicas, físico-químicas e biológica da água dos tanques de cultivo e de depuração. Os peixes provenientes de área de cultivo do Estado de São Paulo, região de Artur Nogueira, da espécie tilápia do Nilo, Oreochromis niloticus, foram submetidos à depuração (3, 5 e 7 dias) e a seguir foram eviscerados e filetados. Os peixes submetidos à depuração e o controle foram comparados entre si para posterior escolha do melhor tempo para depuração; estes, não apresentaram diferenças significativas (p>0,05) para composição centesimal, nitrogênio não protéico - NNP, bases nitrogenadas voláteis totais - BNVT e pH nos tempos de depuração. O processo de depuração provocou perda de peso mais intensa nos três primeiros dias de depuração. Através das análises sensoriais verificou-se que os peixes, que corresponderam ao controle, apresentaram maior intensidade de off flavor quando comparados aos demais. A depuração por 3 dias para o atributo odor apresentou eficiência semelhante, estatisticamente (Teste F), ao tratamento por 5 dias porém, foi considerada diferente do tratamento por 7 dias. Para o atributo sabor os tratamentos de 3, 5 e 7 dias não diferiram, estatisticamente, entre si. A água do tanque de cultivo apresentou problemas de eutrofização e presença de geosmina, ao contrário da água do tanque de depuração que apresentou-se límpida e ausente de geosmina. Os peixes que não sofreram depuração (controle), foram submetidos à defumação. Os filés de peixe in natura e os defumados foram comparados entre si, quanto a composição centesimal e verificou-se que estes foram afetados significativamente (Teste F). Para o nitrogênio não protéico - NNP, as bases nitrogenadas voláteis totais - BNVT e o pH não houve alteração significativa (p>0,05) quanto aos tratamentos. Através das análises sensoriais verificou-se que os peixes in natura apresentaram maior intensidade de off flavor quando comparados aos defumados. Para os atributos odor e sabor houve diferença significativa (p>0,05) entre os tratamentos. O processo de depuração permitiu bons resultados quanto a eliminação de off flavor da tilápia, pois, através das análises realizadas, pode-se aferir o tempo ideal de 5 dias de depuração conduzida em um tanque com água corrente e limpa. Quando se comparou o pescado defumado com o pescado in natura, o primeiro recebeu uma maior aceitação pelos degustadores. O processo de defumação é uma forma de mascarar a presença do off flavor; é um processo simples, pouco oneroso e que pode ser adotado prontamente pelos produtores como forma de agregar valor ao pescado.
Currently, economic losses are a reality due to the fish off flavor provoked by the handling used in the culture and by the water quality of the fish tanks, which are not always satisfactory. Microbiological, physical-chemical and sensory evaluations of the fish were carried out in order to detect probable alterations that would compromise the fish quality. The off flavor detection, as well as, the suggestion for depuration before distribution and sale, and the use of the smoking as a way of disguising this problem were the objectives of this research. Microbiological, physical-chemical and biological analyses of the waters in the fish and the depuration ponds were performed to monitor the quality of the fish. The fish– Nile Tilapias (Oreochromis niloticus) from the area of Arthur Nogueira, in the State of São Paulo – was submitted to the depuration (3, 5 and 7 days). The fish submitted to the depuration and the control fish were compared in order to find the best depuration period. No significant differences (p>0.05) for centesimal composition, non-protein nitrogen - NNP, total base volatile nitrogen BNVT, and pH were found among the depuration periods. The depuration process provoked a more intense weight loss in the first three days for depuration. Through the sensory analyses, we could verify that the control fish presented a greater off flavor intensity than the others. The three-day depuration showed a similar statistical efficiency (F Test) than that of the five-day treatment, but differed from that of the 7-day treatment. As for the flavor attribute, the 3, 5, and 7-day treatments did not show a significant difference, statistically. The water in the fish pond presented eutrophization problems and presence of geosmine, whereas the water in the depuration pond was clear and geosmine-free. The fish that did not go through depuration (control) were submitted to smoking. Fresh fish fillets were compared to the smoked ones. As to the centesimal composition, we verified that they were significantly affected (F Test). As to non-protein nitrogen – NNP, total base volatile nitrogen – BNVT and pH, there were no significant changes (p>0.05) either among the treatments. Through sensory analysis, we verified that the fresh fish presented a more intense off flavor than the smoked ones. As to odor and flavor, there was a significant (p>0.05) difference among the treatments. The depuration process was efficient in the elimination of the off-flavor in the tilapia, for, according to our analyses, we could conclude that the ideal period for this elimination was 5 days of depuration, carried out in a pond with clean running water. When compared to the fresh fish, the smoked fish was better accepted by the panelists. The smoking process is a way of disguising the off flavor; it is a simple, inexpensive process, which can be readily adopted by producers a way of adding value to the fish.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
A irradiação é uma técnica de conservação ainda pouco aceita para alimentos e bebidas no Brasil, devido a grande parte da população desconhecer seus mecanismos de ação. Dessa forma, o objetivo desse estudo foi avaliar os aspectos físico-químico, bioquímico, microbiológico e sensorial de bebida mista de extrato hidrossolúvel de soja (EHS) e suco de uva, submetida à radiação gama. Neste estudo, foram realizados quatro experimentos, visando selecionar o melhor cultivar de soja (BRS-213, BRS-258 e EMBRAPA-48) em relação às características bioativas após a aplicação de diferentes doses de radiação gama (experimento 1). No segundo experimento, selecionou-se o método mais apropriado de produção do EHS a partir de duas formas de extração; determinou-se também a dose de radiação adequada para se aplicar ao EHS (etapa 1) e ao suco de uva (etapa 2), no experimento 3. No experimento 4 foi determinada a temperatura ideal de armazenamento e a adição ou não do conservante ácido benzoico em bebida mista de EHS e suco de uva. Para o experimento 1, realizou-se análises bioquímicas, sendo que nos demais, foram realizadas análises físico-químicas, bioquímicas, microbiológicas e sensoriais. No experimento 4, realizou-se também o teste de vida de prateleira. A partir dos resultados submetidos à estatística descritiva e inferencial (testes paramétricos - t e ANOVA; e testes não paramétricos -Kruskall-Wallis e Friedmann) constatou-se que a elevação da dose de radiação gama aumentou o teor dos compostos fenólicos para os cultivares de soja BRS-213 e BRS-258, bem como a atividade da enzima superóxido dismutase no cultivar BRS-213 (experimento 1). O EHS extraído a frio apresentou maior teor de proteína, lipídios e compostos fenólicos que o EHS extraído a quente. Por outro lado, a segunda extração apresentou melhores resultados na análise sensorial (experimento 2). No experimento 3 o EHS (etapa 1) submetido a 8 kGy apresentou menor teor de proteína e de compostos fenólicos. O EHS e o suco de uva (etapa 2) submetidos a 8 kGy demonstraram variação na cor e redução das notas do atributo sabor quando comparados aos demais tratamentos. A partir da dose de 2 kGy eliminou-se os bolores e leveduras do suco de uva. A bebida mista refrigerada e com adição de ácido benzoico apresentou maior vida de prateleira (9 meses). Concluiu-se que a dose de 4 kGy associada a refrigeração e ao conservante foi indicada para bebida mista de EHS com suco de uva, preservando suas características fitossanitárias, físico-químicas, bioquímicas e sensoriais.
The gamma irradiation is a conservation technique still not accepted for food and beverages in Brazil, due to much of the population ignore their mechanisms of action. Thus, the aim of this study was to evaluate the physic-chemical, biochemical, microbiological and sensory aspect of mixed beverage of soybean hydrosoluble extract and grape juice, submitted to gamma radiation. In this study, four experiments were conducted in order to select the best soybean cultivar regarding the bioactive characteristics (BRS-213, BRS 258 and EMBRAPA-48) after application of different doses of gamma radiation (experiment 1). In the second experiment, we selected the most appropriate method of producing soybean soybean hydrosoluble extract (EHS) from two forms of extraction. Also determined the adequate dosage of radiation to be applied to the EHS (step 1) and grape juice (step 2), in experiment 3. And the experiment 4 was determined the ideal storage temperature and with or without addition of the preservative benzoic acid in the mixed beverage of EHS and grape juice. For the experiment 1 was performed biochemical analyzes, and in the other were conducted in physical-chemical, biochemical, microbiological and sensory analiyses. In experiment 4, it was also carried shelf life test. When results submitted to descriptive and inferential statistics (tests parametric- t and ANOVA, and nonparametric tests -Kruskall-Wallis and Friedman) it was found that the elevation of gamma radiation dose increased the content of phenolic compounds for the soybean cultivars BRS-213 and BRS-258 and SOD activity in cultivar BRS-213 (experiment 1). The EHS cold extraction has a higher protein, lipid and phenolic compounds content that the EHS hot extraction. On the other hand, the second extraction showed better results in sensory analysis (experiment 2). In experiment 3 EHS (step 1) submitted to 8 kGy showed lower protein and phenolic compounds content. The EHS and grape juice (step 2) submitted to 8 kGy showed variation in color and reduction of notes of flavor attribute when compared to other treatments. From the dose of 2 kGy eliminates molds and yeast of grape juice. The refrigerated mixed beverage and with the addition of benzoic acid had longer shelf life (9 months). It was concluded that the dose of 4 kGy associated with refrigeration and preservative was nominated for mixed beverage of EHS with grape juice, preserving their phytosanitary characteristics, physical-chemical, biochemical and sensory.

Os alimentos funcionais suplementados com microrganismos probióticos e ingredientes prebióticos se destacam por seus impactos positivos sobre a saúde do consumidor, representando uma categoria em ascensão: os produtos alimentícios simbióticos. O presente trabalho objetivou desenvolver um produto alimentício, para consumo em porções individuais, a partir da associação de uma barra de cereais a um gelado comestível com baixo teor de gordura e adicionado dos microrganismos probióticos LactobaciIlus acidophilus La-5 e Bifidobacterium animalis Bb-12, suplementado ou não com o ingrediente prebiótico inulina, verificar a viabilidade dos probióticos, avaliar a aceitabilidade sensorial do produto e suas características físico-químicas durante o seu armazenamento a -18°C. Utilizando um planejamento fatorial 22, foram produzidos (em triplicata) quatro tratamentos da porção gelado comestível, todos adicionados de probióticos: T1 (controle), T2 (adição de inulina), T3 (teor reduzido de gorduras) e T4 (adição de inulina e teor reduzido de gorduras). Os produtos foram armazenados a -18°C por até 168 dias. Os parâmetros avaliados na porção gelado comestível foram: pH (antes e após a maturação da calda e após 168 dias de armazenamento), Overrun (após o congelamento), viabilidade dos probióticos (na mistura final e após 1, 2, 7, 14, 21, 28, 84 e 168 dias), dureza instrumental (texturômetro TA-XT2) e fração de derretimento (após 14 dias). Os quatro tratamentos do produto final (barra de cereal em conjunto com a porção gelado comestível) foram avaliados sensorialmente por provadores não treinados, após 7, 28 e 84 dias de armazenamento, utilizando-se teste de aceitabilidade, com escala estruturada de nove pontos. As análises para determinação da composição centesimal da porção gelado comestível e da barra de cereais foram realizadas, para cada porção individualmente. As populações de L. acidophilus e B. animalis foram superiores a 7 log UFC/g, por até 168 dias nas formulações suplementadas com inulina e/ou substituto de gordura e é adição de inulina contribuiu para a manutenção da viabilidade de B. animalis ao longe do armazenamento. A adição e/ou substituição de ingredientes da formulação de gelado comestível não interferiu nos parâmetros físico-químicos pH e overrun. Para a dureza e a velocidade de derretimento, foi verificada diferença significativa (p<0,05). Entretanto, tais diferenças não se refletiram na aceitação do produto pelo consumidor uma vez que não foi verificada diferença para a aceitabilidade sensorial entre o: quatro tratamentos avaliados, com notas sempre superiores a 7 e sem a interferência: do período de armazenamento sobre essa aceitabilidade. O presente trabalho mostrou que a associação de uma barra de cereais a um gelado comestível probiótico não fermentado, com 1,5% de gordura láctea e adicionado de 8% de inulina, tecnologicamente viável para disponibilizar ao consumidor uma alternativa de alimento funcional simbiótico para consumo em porções individuais.
Functional foods supplemented with probiotic microorganisms and prebiotic ingredients are increasingly popular as they improve consumer health, and these foods nowadays form a new group of food products called synbiotic products. The present study aimed to develop a food product for consumption in individual portions, associating a cereal bar and a low fat ice cream bar containing the probiotic microorganisms Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12, supplemented or not with the prebiotic ingredient inulin, to verify the viability of the probiotics and to evaluate the sensory acceptability of the products and their physical-chemical characteristics during storage at -18°C. Employing a 22 design, four trials of the ice cream portion were produced in triplicate, all of them supplemented with probiotics: T1 (control), T2 (with inulin), T3 (with reduced fat content) and T4 (with inulin and reduced fat). The products were stored at -18°C for up to 168 days. The parameters evaluated in the ice cream portion included: pH (before and after the aging of the mix, and after 168 days of storage); overrun (after the freezing stage); viability of the probiotics (in the final mixture and after 1,2, 7, 14, 21, 28, 84, and 168 days); instrumental hardness (TA-XT2 texturometer); and melting rate (after 14 days). The four trials of the final product (cereal bar associated with ice cream bar) were submitted to sensory evaluation by an untrained panel, employing the acceptability test, with a 9-point structured hedonic scale, after 7, 28, and 84 days of storage. The compositional analyses of the ice cream and cereal bars were carried out individually for each portion. The populations of L. acidophilus e B. animalis were above 7 log CFU/g, for up to 168 days for the formulations supplemented with inulin and/or fat substitute, and the addition of inulin contributed to the maintenance of the viability of B. animalis throughout storage. The addition and/or substitution of ingredients in the ice cream formulations did not significantly affect pH and overrun. As for hardness and melting rate, significant differences were observed (p<0.05). However, such differences did not influence the product acceptance by the consumer, as no differences for sensory acceptability between the four trials evaluated was observed, and scores were always above 7, without any interference of the storage period on this acceptability. The study showed that the association of a cereal bar and a low fat non-fermented probiotic ice cream, containing 1.5% milk fat and supplemented with 8.0% inulin, is technologically feasible to provide consumers with a synbiotic functional food product, as an option for consumption of individual portions.

Dissertação de Mestrado em Segurança Alimentar
O desperdício alimentar tem um impacto significativo a nível ambiental, económico e social. Sempre que a sua redução não seja possível, deverá ser promovida a reutilização dos alimentos, através de mercados secundários ou da doação aos membros mais vulneráveis da sociedade. O presente estudo pretendeu caracterizar o processo de redistribuição de refeições e desenvolver um modelo quantitativo de análise do risco relativo associado ao processo. Foi feita uma recolha de dados de temperatura das refeições desde a doação até ao consumo, com recurso a data logger e inquéritos de dados de consumo aos beneficiários. Com base em modelos de crescimento e curvas de dose-resposta disponíveis na literatura, e concentrações iniciais fixas, foi modelado o crescimento de Salmonella, Staphylococcus aureus e Listeria monocytogenes e calculados os riscos relativos nos momentos da recolha da refeição e do consumo. O risco relativo médio foi de 1,30; 1,27 e 1,02 na recolha e 1,25; 1,57 e 1,04 no 1º consumo, respetivamente para Salmonella, S. aureus e L. monocytogenes. Considerando um controlo do processo na sua fase de preparação (máximo 1h a 10ºC), foi possível reduzir o risco relativo na recolha para 1,06; 1,11 e 1,0. O risco associado ao processo de reaproveitamento de refeições pode ser reduzido através de um maior controlo das temperaturas de conservação nas instituições mediadoras e da sensibilização dos beneficiários para a melhoria da conservação e para a importância do consumo dentro do prazo estabelecido.
ABSTRACT - Relative Risk Assessment of meals redistribution for human consumption - Case study - Food waste has a significant environmental, economic and social impact. When it´s not possible to reduce this waste, re-use of food through secondary markets or giving to the most vulnerable members of society should be promoted. The present study aimed to characterize the process of reuse of meals and to develop a quantitative model of relative risk analysis associated to the process. Meal temperature data were collected from donation to consumption using data logger and consumer data surveys. Based on growth models and dose-response curves available in the literature, and fixed initial concentrations, the growth of Salmonella, Stapylococcus aureus and Listeria monocytogenes was modeled and relative risks were calculated at times of meal collection and consumption. The mean relative risk was 1.30; 1.27 and 1.02 at collection and 1.25; 1.57 and 1.04 at the 1st consumption, respectively for Salmonella, S. aureus and L. monocytogenes. Considering a control of the process in its preparation phase (maximum 1h at 10ºC), it was possible to reduce the relative risk at the collection to 1.06; 1.11 and 1.0. The risk associated with the meal reuse process can be reduced through a better control of the storage temperatures in the mediating institutions and the awareness of the beneficiaries to improve the conservation and consumption within the established period.
N/A

O Brasil consolidou-se como o principal produtor e exportador mundial de carne bovina. Estudos microbiológicos, geralmente realizados com amostras de carne coletadas no comércio e não na cadeia produtiva de carne, resultam numa insuficiência de dados a respeito das características fenotípicas e genotípicas das bactérias patogênicas de relevância nos produtos destinados à exportação. Objetivando determinar a prevalência e características de Salmonella spp em carne bovina para exportação, realizou-se a coleta de amostras de superfícies de 200 bovinos adultos, provenientes de 12 fazendas, abatidos em Frigorífico Exportador em São Paulo, Brasil, no ano de 2008. Foram coletadas amostras do couro do animal, logo após a realização da sangria (Co), da carcaça do mesmo animal, após a esfola (Ca I) e da carcaça do mesmo animal, após o toalete e antes da refrigeração (Ca II). O isolamento e a identificação de Salmonella spp foram realizados de acordo com o método - ISO 6579:2002, com algumas modificações. O patógeno foi detectado em 14 amostras de couro (7,0%), 5 de carcaça I (2,5%) e 4 de carcaça II (2,0%). Verificou-se a prevalência do sorovar S. Give (52,0%), seguido de S. Abaetetuba (16,0%), S. Typhimurium (8,0%), S. Agona (4,0%) e S. Dublin (4,0%), e quatro cepas (16,0%) não tipáveis. A tipagem molecular, feita por PFGE, mostrou que as salmonelas expressaram 12 perfis genéticos distintos, sendo 10 formados por apenas uma cepa, cada. As demais cepas (15) pertenceram a dois perfis genéticos apenas, que apresentaram 91,7% de similaridade. De acordo com o teste de infecção de células Caco-2, a maioria das cepas (92,0%) apresentou Eficiência de Invasão inferior a 1,0%, indicando baixo potencial de virulência. Quanto ao perfil de resistência a antibióticos, 68,0% das cepas analisadas foram multiresistentes, apresentando 12 perfis diferentes. Animais diferentes, provenientes de uma mesma fazenda, apresentaram salmonelas de um mesmo sorovar e com o mesmo perfil genético e de resistência a drogas, comprovando a ocorrência de contaminação cruzada durante o processamento da carne bovina. A multiresistência das salmonelas isoladas e a possibilidade de disseminação desses patógenos denotam a necessidade de se adotar medidas de higiene adequadas e maior prudência no emprego de antimicrobianos, na dieta alimentar e na terapêutica veterinária.
Brazil is an important bovine meat producer and exporter. Microbiological surveys are generally run with meat samples collected at retail level and not with meat for export, explaining the lack of data on the phenotypic and genotypic characteristics of pathogens of relevance in exported food products. This study aimed to evaluate the prevalence and characteristics of Salmonella spp in bovine meat destined for export, through surface sampling of hides and carcasses of 200 animals, from 12 farms, slaughtered in 2008 in an export slaughterhouse located in São Paulo, Brazil. Sampling was done from the hides right after bleeding (Co) and from carcasses of the same animal after removal of the hide (Ca I) and after cleaning but before chilling (Ca II). Isolation and identification of Salmonella spp were done according to ISO 6579:2002, with some modifications. The pathogen was detected in 14 samples of hides (7,0%), 5 of carcasses Ca I (2,5%) and 4 of carcasses Ca II (2,0%). The most prevalent serovars were S. Give (52,0%), followed by S. Abaetetuba (16,0%), S. Typhimurium (8,0%), S. Agona (4,0%) and S. Dublin (4,0%). Four isolates (16,0%) were not typable. Molecular typing using PFGE indicated that the isolates presented 12 molecular profiles, ten of them containing one single isolate. Fifteen isolates belonged to only two distinct profiles, with 91.7% similarity. Invasion Efficiency tests, run with Caco-2 cells, indicated that most isolates (92,0%) presented low virulence potential. 68,0% of the isolates were multiresistant to antimicrobial drugs, presenting 12 different resistance profiles. Different animals, coming from the same rearing farm, harbored salmonellae belonging to same serovar and presenting the same genetic and antimicrobial resistance profiles, indicating cross contamination in the slaughterhouse during production of meat. The occurrence of salmonellae that can disseminate in the slaughterhouse and the multiresistance presented by the strains strengthen the need for adoption of proper hygiene control measures and care in the use of antibiotics in human and veterinary therapeutics.

O aumento do consumo de vegetais frescos e a globalização do mercado de hortaliças e frutas frescas provocaram um aumento na preocupação com as enfermidades transmitidas por alimentos (ETA) associadas a esses produtos. No Brasil, a produção de hortaliças orgãnicas vem crescendo, aproximadamente 40% ao ano. Considerando o exposto, foram analisadas 108 amostras de agrião orgãnico minimamente processado e irradiado coletadas, aleatoriamente, em produtores da região de São Roque, São Paulo, no período de novembro de 2005 a março de 2007, para avaliar a ecologia microbiana e a concentração de vitamina C ao longo da cadeia produtiva. As amostras de agrião orgãnico coletadas no campo e as de minimamente processado apresentaram populações superiores a 3,0 log UFC/g para aeróbios mesófilos, aeróbios psicrotróficos, Pseudomonas spp, coliformes termotolerantes e E. coli. Salmonella spp, E. coli 0157:H7 e L. monocytogenes não foram detectadas ao longo da cadeia produtiva. Comparando o processo mínimo com a combinação processo mínimo e irradiação constata-se que a combinação é mais eficiente uma vez que o processo mínimo seguido de exposição à dose de 1 kGy foi suficiente para reduzir de maneira significativa os diversos grupos de mícrorganismos no agrião. As concentrações de ácido ascórbico, ácido dehidroáscorbico e de vitamina C variaram em todas as etapas de processamento do agrião orgãnico minimamente processado e irradiado. Ao longo da vida-de-prateleira de agrião orgãnico minimamente processado e irradiado, a população de L. monocytogenes foi reduzida em, aproximadamente, 4,5, 5,5 e 5,9 ciclos log de acordo com as doses de 1 kGy, 2 kGye 3 kGy, respectivamente. Comportamento similar pode ser constatado para a população de Salmonella spp,. Os resultados da análise sensorial mostraram que o conhecimento ou não do processo de irradiação pelo consumidor não prejudica a aceitação e a intenção de compra do produto irradiado. Portanto o processo de irradiação visando a melhoria da qualidade de agrião orgânico é factível desde que sejam seguidas as Boas Práticas Agrícolas, Boas Práticas de Produção e Boas Práticas do Processo de Irradiação.
With the increase in the consumption of fresh vegetables and the globalization of the market for fresh fruits and vegetables, the concern with foodborne diseases associated with these products has also increased. In Brazil, the production of organic vegetables has increased approximately 40% per year in the last decade. Considering the above, 108 samples of irradiated, minimally processed, organic watercress from producers in the region of Sao Roque, Sao Paulo, were collected through November 2005 to March 2007, to assess the microbial ecology and vitamin C content. Samples of organic watercress collected at the farm level and at the industry level showed populations higher than 3,0 UFC/g for aerobic mesophilic, psychrotrophic, Pseudomonas spp, fecal coliforms and E. coli. Salmonella, E. coli O157:H7 and L. monocytogenes were not detected along the production chain. Comparing the minimal process and the combination of minimal process and irradiation, the combination was more efficient since even the lower dose (1 kGy) was sufficient to reduce the population of the various groups of microorganisms. The ascorbic acid, dehidroascorbic acid and vitamin C content varied at all stages of minimal processing as well as with the exposed doses of irradiation (1, 2 and 3 kGy). The population of L. monocytogenes decreased approximately 4.5, 5.5, and 5.9 log cycles, depending on the exposed doses, throughout the shelf life of irradiated minimally processed organic watercress. Similar behavior was showed by the Salmonella population. Sensory evaluation results showed that previous knowledge or none by consumers does not interfere with the acceptance and intention of purchase of irradiated minimally processed organic watercress. Thus the process of irradiation to improve the microbiological quality of minimally processed organic watercress is feasible provided that Good Agricultural Practice, Good Practice Production and Good Irradiation Practice are followed.

Equine protozoal myeloencephalitis (EPM), caused by the protozoan parasite Sarcocystis neurona, is one of the most important neurological diseases of horses in the Americas. While seroprevalence of S. neurona in horses is high, clinical manifestation of EPM occurs in less than 1% of infected horses. Factors governing the occurrence and severity of EPM are largely unknown, although horse immunity might play an important role in clinical outcome. We hypothesize that EPM occurs due to an aberrant immune response, which will be discernable in the equine IgG subisotypes a, b, and (T) that recognize S. neurona in infected diseased horses versus infected but clinically healthy horses. Based on previously-established serum antibody concentrations for IgG subisotypes in healthy horses, standard curves were generated and served to establish the concentration of antigen-specific IgG subisotypes in equine serum and CSF in infected diseased and infected normal horses. The subisotype concentrations and ratios between subisotypes were analyzed to assess whether neurological disease is associated with detectable differences in the antibody response elicited by infection. Results indicate a type I biased immune response in infected diseased horses, implicating the role of immunity in the development of EPM.

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.

Doutoramento em Biologia
Rapid and specific detection of foodborne bacteria that can cause food spoilage or illness associated to its consumption is an increasingly important task in food industry. Bacterial detection, identification, and classification are generally performed using traditional methods based on biochemical or serological tests and the molecular methods based on DNA or RNA fingerprints. However, these methodologies are expensive, time consuming and laborious. Infrared spectroscopy is a reliable, rapid, and economic technique which could be explored as a tool for bacterial analysis in the food industry. In this thesis it was evaluated the potential of IR spectroscopy to study the bacterial quality of foods. In Chapter 2, it was developed a calibration model that successfully allowed to predict the bacterial concentration of naturally contaminated cooked ham samples kept at refrigeration temperature during 8 days. In this part, it was developed the methodology that allowed the best reproducibility of spectra from bacteria colonies with minimal sample preparation, which was used in the subsequent work. Several attempts trying different resolutions and number of scans in the IR were made. A spectral resolution of 4 cm-1, with 32 scans were the settings that allowed the best results. Subsequently, in Chapter 3, it was made an attempt to identify 22 different foodborne bacterial genera/species using IR spectroscopy coupled with multivariate analysis. The principal component analysis, used as an exploratory technique, allowed to form distinct groups, each one corresponding to a different genus, in most of the cases. Then, a hierarchical cluster analysis was performed to further analyse the group formation and the possibility of distinction between species of the same bacterial genus. It was observed that IR spectroscopy not only is suitable to the distinction of the different genera, but also to differentiate species of the same genus, with the simultaneous use of principal component analysis and cluster analysis techniques. The utilization of IR spectroscopy and multivariate statistical analysis were also investigated in Chapter 4, in order to confirm the presence of Listeria monocytogenes and Salmonella spp. isolated from contaminated foods, after growth in selective medium. This would allow to substitute the traditional biochemical and serological methods that are used to confirm these pathogens and that delay the obtainment of the results up to 2 days. The obtained results allowed the distinction of 3 different Listeria species and the distinction of Salmonella spp. from other bacteria that can be mistaken with them. Finally, in chapter 5, high pressure processing, an emerging methodology that permits to produce microbiologically safe foods and extend their shelf-life, was applied to 12 foodborne bacteria to determine their resistance and the effects of pressure in cells. A treatment of 300 MPa, during 15 minutes at room temperature was applied. Gram-negative bacteria were inactivated to undetectable levels and Gram-positive showed different resistances. Bacillus cereus and Staphylococcus aureus decreased only 2 logs and Listeria innocua decreased about 5 logs. IR spectroscopy was performed in bacterial colonies before and after HPP in order to investigate the alterations of the cellular compounds. It was found that high pressure alters bands assigned to some cellular components as proteins, lipids, oligopolysaccharides, phosphate groups from the cell wall and nucleic acids, suggesting disruption of the cell envelopes. In this work, bacterial quantification and classification, as well as assessment of cellular compounds modification with high pressure processing were successfully performed. Taking this into account, it was showed that IR spectroscopy is a very promising technique to analyse bacteria in a simple and inexpensive manner.
A deteção rápida e específica de bactérias que podem provocar deterioração de alimentos ou doenças associadas ao seu consumo é cada vez mais importante na indústria alimentar. A deteção, identificação e classificação de bactérias são geralmente realizadas utilizando métodos tradicionais baseados em testes bioquímicos e/ou serológicos e em métodos moleculares baseados em análise de DNA ou RNA. Contudo, estas metodologias são dispendiosas, demoradas e trabalhosas. A espectroscopia de infravermelho é uma técnica confiável, rápida e económica, que pode ser explorada como ferramenta para a indústria alimentar. Nesta tese foi avaliado o potencial da espectroscopia de infravermelho para estudar a qualidade bacteriana de alimentos. No capítulo 2, foi desenvolvido um modelo de calibração que permitiu prever com sucesso a concentração bacteriana de fiambre naturalmente contaminado, mantido em refrigeração durante 8 dias. Nesta parte, foi desenvolvida a metodologia que permitiu obter a melhor reprodutibilidade dos espectros das colónias de bactérias, com preparação mínima das amostras, que foi utilizada no trabalho subsequente. Foram realizadas várias tentativas para a obtenção de espectros de infravermelho, testando diferentes resoluções e número de scans. Os melhores resultados foram obtidos utilizando uma resolução espectral de 4 cm-1 e 32 varrimentos. De seguida, no capítulo 3, foi feita uma tentativa de identificar 22 bactérias provenientes de alimentos usando a espectroscopia de infravermelho associada a análise multivariada. A análise de componentes principais, utilizada como método exploratório, permitiu a formação de grupos distintos, cada um correspondendo a um género diferente, na grande maioria dos casos. Posteriormente, foi realizada uma análise hierárquica por clusters de forma a investigar a formação de grupos e a possibilidade de distinção de espécies dentro de um mesmo género de bactérias. Observou-se que a espectroscopia de infravermelho é adequada não só para a distinção de diferentes géneros, mas também para diferenciar espécies dentro de um mesmo género, com o uso simultâneo de análise de componentes principais e análise hierárquica por clusters. A utilização de espectroscopia de infravermelho e análise estatística multivariada foram também investigadas no capítulo 4 para confirmação da presença de Listeria monocytogenes e Salmonella spp., isoladas a partir de alimentos contaminados, após crescimento em meio selectivo. Isto permitiria a substituição dos métodos bioquímicos e serológicos que são usados para confirmar a presença destas bactérias patogénicas e que podem atrasar a obtenção de resultados por 2 dias. Os resultados obtidos permitiram a distinção de Salmonella spp. de outras bactérias que se possam confundir com elas. Por fim, no capítulo 5, o processamento por alta pressão, uma metodologia emergente que permite produzir alimentos microbiologicamente seguros e aumentar o seu tempo de prateleira, foi aplicada a 12 bactérias alimentares, de forma a determinar a sua resistência e os efeitos da pressão a nível das células. Foi aplicado um tratamento de 300 MPa, à temperatura ambiente e durante 15 minutos. As bactérias de Gram-negativo foram inativadas até níveis não detetáveis, enquanto as de Gram-positivo mostraram diferentes níveis de resistência. As espécies Bacillus cereus e Staphyloccus aureus decresceram apenas 2 unidades logarítmicas enquanto a espécie Listeria innocua diminuiu cerca de 5 unidades logarítmicas. A espectroscopia de infravermelho foi utilizada na análise das colónias bacterianas antes e após o tratamento por alta pressão, de forma a investigar as alterações que são provocadas nos componentes celulares com este tipo de processamento. Descobriu-se que a alta pressão altera bandas espectrais correspondentes a alguns componentes celulares, de entre os quais proteínas, lípidos, oligopolissacarídeos, grupos fosfato da parede celular e ácidos nucleicos, podendo indicar rutura da parede/membrana celular. Neste trabalho, a quantificação de bactérias e a sua classificação, bem como a análise de modificação nos componentes celulares após processamento por alta pressão foram realizados com sucesso. Assim, a espectroscopia de infravermelho demonstrou ser uma técnica bastante promissora para analisar bactérias provenientes de alimentos de uma forma simples e pouco dispendiosa.

O presente trabalho visou o desenvolvimento de uma sobremesa aerada simbiótica tipo musse, com baixo teor de gordura, processada com a adição da cultura probiótica de Lactobacillus acidophilus La-5, dos ingredientes prebióticos oligofrutose e inulina e de concentrado protéico de soro de leite (WPC), para ser armazenada sob refrigeração e congelamento, e a comparação do efeito desses ingredientes sobre as características do produto e a resistência in vitro do probiótico adicionado. Utilizando o delineamento experimental para misturas de três fatores e um ponto central, 7 formulações de musses de goiaba foram estudadas durante o armazenamento a 4°C, durante 28 dias, e a -18°C, durante 112 dias. As maiores populações de L. acidophilus foram alcançadas nas musses congeladas, com valores sempre superiores a 7 log UFC/g por até 12 semanas de armazenamento a -18°C. A viabilidade de L. acidophilus se mostrou satisfatória até o 28º dia nas musses refrigeradas adicionadas de WPC, com populações variando entre 7,7 e 6,2 log UFC/g. Para as demais musses, a população do probiótico chegou a reduzir, em média, até 2 log após 28 dias. L. acidophilus apresentou grande redução da viabilidade, tanto para as musses refrigeradas como para as congeladas, nos ensaios de sobrevivência às condições gastrintestinais simuladas in vitro. Nas musses refrigeradas, a substituição total ou parcial da gordura láctea por inulina resultou em melhor sobrevivência do probiótico durante o ensaio in vitro na primeira semana. Ao considerar o período completo de armazenamento, a menor redução da viabilidade de L. acidophilus ao longo dos ensaios in vitro foi observada para as musses congeladas. A substituição da gordura láctea por inulina e WPC e o congelamento resultaram em diferenças significativas nos parâmetros de textura das musses (p<0,05), não interferindo, porém, na sua aceitabilidade sensorial. Considerando as populações máximas do probiótico ao longo do armazenamento e a sua sobrevivência às condições gastrintestinais simuladas in vitro, observou-se os melhores resultados com a substituição parcial da gordura láctea adicionada no produto refrigerado por WPC, na proporção de 2 a 3% da formulação. A adição simultânea de WPC e inulina para musses armazenadas sob refrigeração e congelamento é recomendada desde que a proporção conjunta desses ingredientes não ultrapasse 2,6%, no sentido de não prejudicar a textura e as características sensoriais do produto. De modo especial para as musses congeladas, também é aconselhada a proporção de 2% de gordura láctea e 2% de inulina, para a qual foram obtidos os melhores resultados nos ensaios de sobrevivência in vitro do probiótico.
The aim of the present study was to develop a mousse-type synbiotic aerated dessert with low fat content, supplemented with the Lactobacillus acidophilus La-5 probiotic culture, the prebiotic ingredients oligofructose and inulin and whey protein concentrate (WPC), to be stored refrigerated and frozen, and to compare the effect of these ingredients on the product characteristics and the in vitro resistance of the added probiotic microorganism. Using a simplex centroid design, seven guava mousse-making trials were studied during storage at 4°C for 28 days and at -18°C for 112 days. The highest populations of L. acidophilus were achieved in frozen products, always above 7 log CFU/g at up to 12 weeks of storage at -18°C. L. acidophilus viability was satisfactory up to 28 days in the refrigerated mousses supplemented with WPC, with populations between 6.2 and 7.7 log CFU/g. For the other mousses, the populations decreased around 2 log cycles afier 28 days of refrigerated storage. Reductions in L. acidophilus survival during the in vitro assays were high both for refrigerated and frozen mousses. For the refrigerated mousses, the total or partial substitution of milk fat by inulin increased the probiotic survival during the in vitro assays in the first week. In terms of the whole storage, L. acidophilus survival decreased less during the in vitro assays for the frozen mousses. The substitution of milk fat by inulin and WPC and the frozen storage lead to significant differences in the texture of mousses (p<0.05), without affecting their sensory acceptability. Considering the maximum probiotic populations during storage and the survival under the in vitro simulated gastrointestinal conditions, the best results for the refrigerated product were obtained with the partial substitution of milk fat by WPC at 2 to 3%. The simultaneous addition of inulin and WPC is recommended. However, the total proportion of both ingredients together should not exceed 2.6%, so as to obtain a texture profile and a sensory acceptance similar to the traditional product. Particularly for frozen mousses, the mixture of 2% milk fat and 2% inulin in the formulation is also suggested, for which the best results on probiotic survival in the in vitro assays were observed.

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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.