Made available in DSpace on 2017-07-10T17:44:40Z (GMT). No. of bitstreams: 1 Michelle_Medeiros_Carvalho.pdf: 938080 bytes, checksum: 942326b46b04f46650ea2a86d579a566 (MD5) Previous issue date: 2015-09-21 / In the name of food policy, the processing and marketing of Artisanal Cheese made from raw milk have been inhibited by the action of regulatory agencies, which have been demanding pasteurization of milk for the manufacture of cheese as a solution to ensure product safety. A specific legislation for raw milk cheese production was recently published to meet various requirements of society in defense of Artisanal Cheese. The aim of this study was to investigate the situation of 12 rural properties in Seara-SC that were producing and selling Artisanal Colonial-Type Cheese informally. Producers were visited and their conditions for obtaining water, performing milking and making cheese were assessed. A semi-structured questionnaire was used to analyze the difficulties faced by farmers in compliance with current legislation for artisanal raw milk cheese production. Physical, chemical and microbiological analyzes of water, milk and cheese were conducted to check the adequacy of legal standards. It was found that none of the Colonial cheese makers investigated met the requirements established by law for Artisanal Cheese production from raw milk. Among the samples of water, milk and cheese analyzed, 91.7% (n = 11), 50% (n = 6) and 100% (n = 12), respectively, were found outside the legal parameters. It was concluded that cheese makers were unable to meet the requirements set by law, without some technical support, because they were difficult to be applied by them. They often need guidance and do not know where to seek orientation to meet all legal specifications. This study also concluded that public policies need to be employed for the preservation of Artisanal Colonial-Type Cheese, considering its historical and socio-economic relevance for family farming, to preserve the cultural tradition of such communities / Em nome da segurança alimentar a elaboração e comercialização de queijos artesanais produzidos a partir de leite cru vem sendo inibida pela ação de órgãos fiscalizadores, que vêm exigindo a pasteurização do leite para o fabrico de queijos como solução para garantir a inocuidade do produto. Para atender a diversas pressões da sociedade em defesa do queijo artesanal, recentemente foi publicada legislação específica para produção de queijo a partir de leite cru. O objetivo deste trabalho foi investigar a situação de 12 propriedades rurais do município de Seara/SC que produziam e comercializavam Queijo Colonial Artesanal de maneira informal. Os produtores foram visitados com o intuito de conhecer as instalações da ordenha e fabricação do queijo, bem como a forma de obtenção e tratamento da água das propriedades. Também foi conhecida a forma como o queijo é produzido. Por meio da aplicação de um questionário semiestruturado, aplicado foram analisadas as dificuldades enfrentadas pelos produtores rurais para atendimento à legislação vigente para a produção de queijo artesanal feito com leite cru. Foram realizadas análises físico-químicas e microbiológicas de água, leite e queijo dessas propriedades rurais para verificação da adequação aos parâmetros legais existentes. Constatou-se que nenhum produtor de Queijo Colonial pesquisado atende às exigências estabelecidas pela legislação em vigor para produção de Queijo Artesanal a partir de leite cru. Das amostras de água, leite e queijo analisadas, 91,7% (n=11), 50% (n=6) e 100% (n=12), respectivamente, encontraram-se fora dos parâmetros legais. Concluiu-se que os produtores não conseguem atender, sem apoio técnico, às exigências estabelecidas pela legislação vigente por se tratar de medidas difíceis de serem aplicadas por eles, que normalmente necessitam de orientação e têm dificuldades no atendimento de todas as especificações legais. O estudo também concluiu que há necessidade de implementação de políticas públicas para a preservação do Queijo Colonial Artesanal, devido à sua importância histórica e socioeconômica dentro da agricultura familiar, perpetuando a tradição cultural desses produtores rurais

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-03-14T17:16:12Z No. of bitstreams: 1 Lopes, Patricia Regina Kraschinski [Dissertação, 2016].pdf: 1353027 bytes, checksum: d3bd0a8823abd0576a16231891f91a13 (MD5) / Made available in DSpace on 2017-03-14T17:16:12Z (GMT). No. of bitstreams: 1 Lopes, Patricia Regina Kraschinski [Dissertação, 2016].pdf: 1353027 bytes, checksum: d3bd0a8823abd0576a16231891f91a13 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo deste estudo foi avaliar a qualidade microbiológica do leite bovino em diferentes etapas da cadeia produtiva de uma cooperativa de leite do Estado do Rio de Janeiro, e investigar a presença de STEC e STEC O157 no leite e nas fezes dos animais ordenhados. Foram coletadas 100 amostras de leite nas diferentes etapas da cadeia produtiva, sendo 54 amostras do galão de cada propriedade rural, 25 amostras de leite do tanque de refrigeração comunitário, 14 amostras dos carros-tanque isotérmicos e 7 amostras do tanque de resfriamento da indústria bem como 10 amostras de leite pasteurizado. Foram também avaliadas 63 amostras de swab retal do animal ordenhado e 35 amostras de leite recém ordenhado. Para conferir a eficiência da pasteurização foram realizados os testes de peroxidase e fosfatase alcalina. A maioria das amostras (75,9%) de leite cru coletadas nos galões dos produtores apresentou condições microbiológicas satisfatórias, contagem bacteriana total (CBT) < 5,5 log UFC/mL. No entanto, 72% das amostras de leite coletadas no tanque de refrigeração comunitário, 100% das amostras coletadas nos carros-tanque isotérmicos e 100% das amostras coletadas nos tanques de refrigeração da indústria apresentaram condições microbiológicas insatisfatórias. Todas as amostras de leite pasteurizado foram aprovadas quanto a CBT, além de apresentarem a enzina peroxidase ativa e a enzima fosfatase alcalina inativa. No entanto, 80% destas amostras foram classificadas em condições microbiológicas insatisfatórias por apresentarem coliformes totais e E. coli acima dos padrões estabelecidos pela legislação brasileira. Das 100 amostras de leite cru analisadas, 65 apresentaram o gene stx, sendo 33 (61,1%) amostras do galão do produtor, 13 (52%) amostras do tanque de refrigeração comunitário, 14 (100%) amostras dos carros-tanque isotérmicos e cinco (71,4%) amostras dos tanques de resfriamento da indústria. Cinquenta e seis (88,9%) amostras fecais e 17 (48,6%) amostras de leite recém-ordenhado apresentaram o gene stx. Com exceção de uma amostra, todas as amostras de leite recém ordenhado provinham de um animal carreador do gene stx. Foram isolados STEC O157:H7 em 11,9% das amostras de fezes bovinas stxpositivas. Todas as cepas STEC O157:H7 isoladas apresentaram os genes stx2c, eae , tir , espA , espB , ler, iha, astA, EHEC-hlyA e espP. A qualidade microbiológica dos produtos lácteos está diretamente relacionada à qualidade da matéria-prima, com isso, é necessário a adequação dos produtores rurais quanto à implementação de Boas Práticas Agropecuárias, além da implementação das Boas Práticas de Fabricação pela indústria de laticínios / The aim of this study was to evaluate the microbiological quality of cow's milk in different stages of the production chain of a cooperative state milk Rio de Janeiro, and to investigate the presence of STEC and STEC O157 on themilk and the faeces of milked animals. 100 milk samples at different stages of the production chain were collected, 54 samples of a gallon of each rural property, 25 milk samples of Community coolant tank, 14 samples of insulated tank cars and 7 samples of the industrial cooling tank; and 10 samples of pasteurized milk. It was also evaluated 63 samples of rectal swab of milking animals and 35 sample of freshly milked milk. To check the effectiveness of the pasteurization tests peroxidase and alkaline phosphatase were performed. The majority of samples (75.9%) of raw milk collected in gallons producers had satisfactory microbiological conditions, total bacterial count (TBC) < 5.5 log CFU/mL. However, 72% of milk samples collected in the Community coolant tank, 100% of the samples collected in the isothermal tank cars and 100% of the samples collected in the industrial cooling tanks had unsatisfactory microbiological conditions. All (100%) samples of pasteurized milk were approved as the TBC, besides having the peroxidase enzyme is of not presenting the enzyme alkaline phosphatase. However, 80% of these samples were classified as unsatisfactory microbiological conditions for presenting total coliforms and E. coli above the standards set by law. Of 100 raw milk samples analyzed, 65 showed the stx gene, 33 (61.1%) of the producer gallon, 13 (52%) of the community coolant tank, 14 (100%) of the isothermal tank cars and five (71.4%) of cooling tanks industry. Fifty-six (88.9%) and 17 faecal samples (48.6%) of freshly milked milk samples showed the stx gene. With the exception of one sample, all milk samples freshly milked, came from a carrier animal stx gene. They were isolated STEC O157: H7 in 11.9% of samples stx-positive cattle dung. All strains STEC O157: H7 isolated genes showed stx2c, eae , tir , espA , espB , read, iha, astA, EHEC-hlyA and espP. The microbiological quality of dairy products is directly related to the quality of the raw material, thus, the adequacy of farmers as the implementation of Best Practices of Agricultural is required in addition to the implementation of Good Manufacturing Practices by the dairy industry

A caseína e os ácidos graxos livres (AGL) são importantes componentes do leite relacionados às suas características industriais e sensoriais. Alguns fatores são responsáveis por alterações em suas concentrações, dentre eles, pode-se destacar as condições de armazenamento, além da agitação e, principalmente, a contagem de células somáticas (CCS). Para estudar tais alterações, avaliou-se através do presente estudo o efeito da temperatura de armazenamento (- 20oC e 7oC) e da idade das amostras (3, 6, 9 e 12 dias), assim como da CCS (CCS1: 400 mil, CCS2:400-750 mil e CCS3: 750 mil células somáticas/mL de leite). O efeito das condições de armazenamento sobre a caseína e os AGL foi avaliado no primeiro estudo. O efeito da CCS sobre a caseína e os AGL, por outro lado, foi avaliado no segundo estudo. No primeiro estudo, observou-se aumento nos teores de AGL ao longo do tempo, ou seja, acompanhando o aumento do período de armazenamento, com médias superiores em amostras resfriadas (7oC), em relação às congeladas (-20oC). Desta forma, sugere-se que análises laboratoriais devem ser realizadas em até três dias para este componente, devido ao seu aumento progressivo em amostras resfriadas. Para a caseína, entretanto, não foi identificada interação entre os fatores idade e temperatura. As médias deste componente não diferiram considerando-se as condições de armazenamento avaliadas, indicando que as análises para este componente podem ser realizadas até doze dias após a coleta, independente da temperatura de armazenamento. No segundo estudo, identificou-se interação entre classes de CCS e época de coleta, para caseína. A partir dos resultados de AGL, pode-se concluir que o leite encontra-se com altas concentrações deste componente, quando coletado na propriedade rural, sendo também observada influência da CCS sobre as concentrações de caseína e AGL no leite cru. Os AGL apresentaram incrementos significativos comparando-se amostras coletadas em diferentes locais, no trajeto da fazenda à indústria. Por outro lado, comparando-se os mesmos locais: tanque, rota e silo, as médias de caseína não diferiram. Observou-se correlação entre as variáveis CCS e caseína e CCS e AGL. / The casein and the free fatty acids (FFA) are important milk components related to its industrial and sensory characteristics. Some factors are responsible for changes in their concentration among them can highlight the storage conditions, as well as agitation and, especially, the somatic cell count (SCC). To study such changes, we assessed through the present study the effect of storage temperature (-20oC and 7oC) and the age of samples (3, 6, 9 e 12 days), as well as the CCS (CCS1: 400 mil; CCS2: 400-750 mil; CCS3: 750 mil somatic cells/mL of milk). The effect of CCS storage conditions on casein and FFA was evaluated in the first study. The effect of CCS on casein and FFA moreover, was evaluated in the second study. In the first study, it was observed increased levels the AGL over time, ie, accompanying the increase of the storage period, with means higher in refrigerated samples (7oC), in relation to frozen samples (- 20oC). Thus, it is suggested that laboratory examinations shall be performed up to three days to this component, due to their progressive increase in refrigerated samples. For casein, however, was not identified interaction between the factors age and temperature. The mean of this component did not differ considering the storage conditions, indicating that the analyzes for this component can be made until twelve days after collection, regardless of storage temperature. In the second study, we identified interactions between CCS classes and seasons, for casein. From the FFA results, we can conclude that milk meets with high concentrations of this component, when collected in the rural property, being also observed influence of CCS on concentrations of casein and FFA in raw milk. The FFA showed significant increases compared to samples collected at different locations on the path from farm to industry. On the other hand, comparing the same locations, tank, route and silo, the casein average did not differ. Correlation was observed between the variables casein and CCS and FFA and CCS.

Bovine milk contains a variety of endogenous peptides, partially formed by milk proteases that may exert diverse bioactive functions. Milk storage allows further protease activities altering the milk peptidome, while processing, e.g., heat treatment can trigger diverse chemical reactions, such as Maillard reactions and oxidations, leading to different posttranslational modifications (PTMs). The influence of processing on the native and modified peptidome was studied by analyzing peptides extracted from raw milk (RM), ultra-high temperature (UHT) milk, and powdered infant formula (IF) by nano reversed-phase liquid chromatography coupled online to electrospray ionization (ESI) tandem mass spectrometry. Only unmodified peptides proposed by two independent software tools were considered as identified. Thus, 801 identified peptides mainly originated from αS- and β-caseins, but also from milk fat globular membrane proteins, such as glycosylation-dependent cell adhesion molecule 1. RM and UHT milk showed comparable unmodified peptide profiles, whereas IF differed mainly due to a higher number of β-casein peptides. When 26 non-enzymatic posttranslational modifications (PTMs) were targeted in the milk peptidomes, 175 modified peptides were identified, i.e., mostly lactosylated and a few hexosylated or oxidized peptides. Most modified peptides originated from αS-caseins. The numbers of lactosylated peptides increased with harsher processing.

Thesis (M. Tech. (Environmental health)) - Central University of technology, Free State, 2013 / Introduction Dairy farms in central South Africa produce a substantial amount of milk, which is sold in Bloemfontein, Free State. Large volumes of unpasteurized (raw) milk is collected on the dairy farms, which undergoes further processing before it reaches the consumer at the end of the production line. There is a large proportion of the population that, in most cases unknowingly, consumes raw milk that has bacterial counts substantially higher than legal standards. Poor quality unpasteurized milk is either sold as fresh milk in the informal market, or as dairy products, such as cheese, manufactured from unpasteurized milk. Consumers are therefore, in most cases, unaware of the poor quality dairy products they consume. Milk quality is usually assessed in terms of bacterial content, which include Escherichia coli, coliforms and total bacterial count. The bacterial quality of milk is influenced by a number of factors, including farming practices, structural design of the milking shed, herd health and quality of water used in the dairy. If the highest level of hygiene practices is maintained, contamination of the milk by pathogenic microorganisms will be controlled, however, any drop in the vigilance of hygiene practices could result in unacceptable high levels of pathogenic microorganisms resulting in poor quality raw milk. Poor quality raw milk will inevitably result in poor quality pasteurized milk, containing unacceptably high levels of pathogenic organisms, which will eventually reach the consumer. Objectives The objectives of this study were to assess the quality of milk and influencing factors of milk produced on 83 dairy farms that supply milk intended for further processing to the greater Mangaung region, Central South Africa. Influencing factors investigated included, water quality and hygiene of milk contact surfaces, namely pulsator surfaces and milk pipeline surfaces. Methods Standard sampling procedures were followed when milk was sampled from bulk milk tanks, water at the point of use in the dairy, as well as collection of surface swabs. Escherichia coli, coliforms, total bacterial counts and somatic cell counts in milk were determined in terms of the regulations relating to milk and dairy products, and for water in terms of drinking water standards. These data were analysed and the factors that directly influence bacterial quality of milk were identified. Results 93% of the dairy farms displayed E. coli in their bulk milk containers, which did not comply with the legal standard. For coliforms, 86% of the milk samples did not comply with the legal standard. The total bacterial count of 85% of the milk samples did comply with the legal standard. The somatic cell count of 42% of the milk samples did not comply with the legal standard. The pulsator surfaces as well as the milk pipeline surfaces of 13% of the dairy farms displayed the presence of E. coli. 80% of the pulsator surfaces and 78% of the milk pipeline surfaces did comply with the legal standard pertaining to coliforms. The total bacterial count of pulsator surfaces revealed that 19% complied, whereas 29% of the milk pipeline surfaces complied with the legal standard. The water data further revealed that 31% of the dairy farms contained E. coli in the water used in the dairies. 63% of the dairy farms contained more than the allowable number of coliforms in their water. Chi-square tests revealed significant differences (p > 0.05) between the presence or absence of E. coli in milk and water; the presence or absence of E. coli in milk and milk pipeline surfaces; the presence or absence of E. coli in milk and pulsator surfaces and the presence or absence of E. coli in milk and the positioning of the cows in the milking shed. When milk quality indexes were calculated for all the farms, only four farms were classified with excellent milk, the remainder were all classified as producing poor quality milk. The hygiene quality indexes revealed that the hygiene practices on all the farms were not up to standard. Discussion and conclusion The study revealed that the milk produced for commercial processing and distribution in the greater Mangaung region of central South Africa was of poor quality. It is often mistakenly believed that the pasteurization process will remove all microorganisms from milk. As this is not the case, it is of major concern that milk delivered commercially is not of acceptable quality. Furthermore, it could be concluded that the quality of milk products from raw milk were also probably not of acceptable quality. The results further revealed that the possible contributing factors to the poor quality milk produced by the 83 commercial dairy farms were; poor quality water used in dairy sheds and contaminated milk contact surfaces. From this study it could be concluded that the overall status of milk production on the 83 commercial dairy farms studied, did not meet the standards required for milk quality, water quality and hygiene practices.

Thesis (M. Tech. (Biomedical Technology)) -- Central University of technology, Free State, 2013 / The objective of this study was to determine the prevalence of Listeria monocytogenes and also to assess the general hygiene of fresh milk in the Maseru area, Lesotho. A total of 200 milk samples (40 pasteurised and 160 raw milk samples) were used for the research. Raw milk samples were collected from the local farmers at the Dairy reception as they bring it for selling. Pasteurised milk samples were bought from different milk selling points in the Maseru area. The total aerobic plate count, total coliform count and total E. coli count for 160 raw milk samples and 40 pasteurised samples were performed to determine the quality of milk. Milk was enriched in selective broths to increase detection sensitivity and was directly plated on selective agars for direct bacterial enumeration. About 54.4% of the of the raw milk samples had total aerobic plate counts greater that 200 000 cfu/ml while 55.6% (89/160) of the raw samples had high counts of greater than 20 cfu/ml for total coliforms, and 21.9% (35/160) of the samples had higher than expected total E. coli counts. High total coliform count was detected in 17.5% (7/40) of the pasteurised milk samples and about 67.5% (27/40) of these samples exceeded the limit for total aerobic plate counts. The counts exceeded the milk standards for pasteurised milk. Phosphatase activity was detected in seven pasteurised milk samples, whereas 33 tested negative for phosphatase activity. Some pasteurised milk samples tested positive for coliform counts which exceeded the maximum limits according to national standards for pasteurised milk. However, most of the pasteurised samples (82.5%) had acceptable counts of less than 20 cfu/ml. API and PCR were used for confirmation and amplification of the isolated Listeria strains. The prevalence of Listeria was found to be (3.75%). Listeria species were found in 6 out of 200 samples tested (160 raw milk samples and 40 pasteurised milk), and were only detected in the raw milk samples. Five species belonged to Listeria monocytogenes and one was Listeria innocua. None of the Listeria was detected in the pasteurised milk samples. Serotyping was done through multiplex PCR with D1, D2, FlaA and GLT primers to determine the serovar groups of L. monocytogenes. All six isolates revealed 214 bp gene which identifies the serotypes in Lineages I or III. The genetic fingerprinting of the isolated Listeria was also determined. Enterobacterial Repetitive Intergenic Consensus (ERIC) sequence-based PCR was used to generate DNA fingerprints with ERIC specific primers. On the basis of ERIC-PCR fingerprints, three different DNA patterns could be discriminated among the analysed isolates. Three L. monocytogenes isolates showed similar DNA banding patterns, while two isolates both had different profiles. A questionnaire was used to determine consumption of raw (unpasteurised) milk or pasteurised milk and its products and it was completed by 300 households from the community. Although there was no indicated prevalence of raw (unpasteurised) milk consumption from the community, participants indicated symptoms alleged to consumption of pasteurised milk and/or milk products. According to community perception some of the dairy products consumed were allegedly implicated in food poisoning illnesses experienced. Participants indicated more symptoms with both fresh and sour milk consumption than in cheese and yogurt consumption.

O gênero Mycobacterium spp. compreende microrganismos saprófitas e patogênicos de interesse em saúde animal e humana. A espécie M. bovis, que causa tuberculose nos animais, é excretada através do leite de bovinos infectados e tem no consumo de leite cru e seus derivados uma importante via de transmissão para o homem, causando uma doença tão grave quanto à causada pelo M. tuberculosis. Como a doença nos animais é endêmica no Brasil e o queijo minas meia cura é normalmente fabricado com leite cru e muito apreciado pelo consumidor paulistano, amostras desse produto, obtidas em feiras-livres, foram analisadas quanto à ocorrência de micobactérias. As amostras foram descontaminadas pelo método HPC 1,5%, semeadas em meio Stonebrink-Leslie (incubadas a 37ºC/90 dias) e as colônias suspeitas, submetidas à reação de PCR TB multiplex e sequenciamento nucleotídico. Em 12% das amostras (16/133) foram isoladas 26 colônias de Mycobacterium spp., tendo sido identificadas 6 espécies, todas ambientais: Mycobacterium fortuitum, M. confluentis, M. elephantis, M. novocastrense, M. sphagni e M. arupense; 7 isolados, no entanto, permaneceram sem caracterização quanto à espécie. O M. fortuitum é um patógeno oportunista importante em saúde pública, sem que haja, entretanto, evidências de transmissão alimentar; o M. novocastrense, M. arupense e M. elephantis também têm sido consideradas espécies com potencial patogênico ao ser humano. Os resultados sugerem, tal como era esperado, que a frequência e a carga inicial de M. bovis em queijo Minas meia cura sejam baixas, mas se deve considerar que a metodologia empregada, por falta de outra específica, não privilegia a detecção em cenário de baixa carga inicial do agente acompanhada por alta carga contaminante. Sugerem também a necessidade de se avaliar a importância da transmissão alimentar de micobactérias não tuberculosas, especialmente para indivíduos imunossuprimidos. / Mycobacterium genus consists of saprophytic and pathogenic microorganisms of interest in animal and human health. M. bovis specie, which causes tuberculosis in animals, is excreted through the milk of infected cattle and the consumption of raw milk and its derivatives is an important route of transmission to humans, causing a disease as serious as the one caused by M. tuberculosis. Considering that the disease in animals is endemic in Brazil and that minas meia cura cheese cure is usually made from raw milk and much appreciated by paulistano consumer, samples of the product acquired in open markets, were analyzed for the occurrence of mycobacteria. The samples were decontaminated by the method HPC 1.5%, sown in Stonebrink-Leslie medium (incubated at 37 ° C/90 da ys) and the suspected colonies, submitted to PCR TB multiplex and nucleotide sequencing reaction. In 12% of samples (16/133) were isolated 26 colonies of Mycobacterium spp. and 6 species have been identified, all of them are environmental Mycobacterium fortuitum, M. confluentis, M. elephantis, M. novocastrense, M. sphagni and M. arupense; 7 isolates, however, remained without characterization for the specie. M. fortuitum is an important opportunistic pathogen in public health, without, however, evidence of being transmitted by food; M. novocastrense, M. arupense and M. elephantis species have also been considered potentially pathogenic to humans. The results suggest that the frequency and/or contamination load of M. bovis in meia cura Minas cheese is (are) low (s). We have to consider, however, that this result may have been influenced by the absence of an analytical method capable of identifying the agent in the food matrix in which a low load of microorganism is expected, accompanied by high load of contaminants. Also suggest the need to evaluate the possible importance of foodborne non-tuberculous mycobacteria, especially for immunocompromised individuals.

The objective of my study was to analyse the influence of selected factors in relation to somatic cell counts (SCC) in bulk milk samples of raw cow´s milk. Milk samples were tested in eight cowsheds with different technology of breeding and milking for a period of three years. The SCC values were determined by the Fluoro-opto-electronic method using the apparatus Fossomatic. A significant factor influencing the SCC was the season. The highest average values of the SCC were found in the summer months. The lowest average values of the SCC were determined in the loose bedded cubicle housing (250.103.ml-1), while the difference in the SCC compared to the loose bedding-free slatted floor housing (SCC average 317.103.ml-1) was highly statistically significant (p < 0,001). In the tie stall with bedding was the SCC average 292.103.ml-1. A highly statistically significant difference in the SCC (p < 0,001) was also demonstrated between breeds in the milking parlour (SCC average 265.103.ml-1) and breeds in the milking stall in the pipeline systems (SCC average 292.103.ml-1). It also showed a statistically highly significant difference in the SCC (p < 0,001) between breeds dominated by the Holstein breed cows (SCC average 285.103.ml-1) and breeds with breed prevalence of the Czech Fleckvieh cows (SCC average 265.103.ml-1). The level of yield was also affected by the SCC. There was no statistically significant difference among the farms using grazing and breeding without any possibility of keeping cows on pasture.

O gênero Mycobacterium spp. compreende microrganismos saprófitas e patogênicos de interesse em saúde animal e humana. A espécie M. bovis, que causa tuberculose nos animais, é excretada através do leite de bovinos infectados e tem no consumo de leite cru e seus derivados uma importante via de transmissão para o homem, causando uma doença tão grave quanto à causada pelo M. tuberculosis. Como a doença nos animais é endêmica no Brasil e o queijo minas meia cura é normalmente fabricado com leite cru e muito apreciado pelo consumidor paulistano, amostras desse produto, obtidas em feiras-livres, foram analisadas quanto à ocorrência de micobactérias. As amostras foram descontaminadas pelo método HPC 1,5%, semeadas em meio Stonebrink-Leslie (incubadas a 37ºC/90 dias) e as colônias suspeitas, submetidas à reação de PCR TB multiplex e sequenciamento nucleotídico. Em 12% das amostras (16/133) foram isoladas 26 colônias de Mycobacterium spp., tendo sido identificadas 6 espécies, todas ambientais: Mycobacterium fortuitum, M. confluentis, M. elephantis, M. novocastrense, M. sphagni e M. arupense; 7 isolados, no entanto, permaneceram sem caracterização quanto à espécie. O M. fortuitum é um patógeno oportunista importante em saúde pública, sem que haja, entretanto, evidências de transmissão alimentar; o M. novocastrense, M. arupense e M. elephantis também têm sido consideradas espécies com potencial patogênico ao ser humano. Os resultados sugerem, tal como era esperado, que a frequência e a carga inicial de M. bovis em queijo Minas meia cura sejam baixas, mas se deve considerar que a metodologia empregada, por falta de outra específica, não privilegia a detecção em cenário de baixa carga inicial do agente acompanhada por alta carga contaminante. Sugerem também a necessidade de se avaliar a importância da transmissão alimentar de micobactérias não tuberculosas, especialmente para indivíduos imunossuprimidos. / Mycobacterium genus consists of saprophytic and pathogenic microorganisms of interest in animal and human health. M. bovis specie, which causes tuberculosis in animals, is excreted through the milk of infected cattle and the consumption of raw milk and its derivatives is an important route of transmission to humans, causing a disease as serious as the one caused by M. tuberculosis. Considering that the disease in animals is endemic in Brazil and that minas meia cura cheese cure is usually made from raw milk and much appreciated by paulistano consumer, samples of the product acquired in open markets, were analyzed for the occurrence of mycobacteria. The samples were decontaminated by the method HPC 1.5%, sown in Stonebrink-Leslie medium (incubated at 37 ° C/90 da ys) and the suspected colonies, submitted to PCR TB multiplex and nucleotide sequencing reaction. In 12% of samples (16/133) were isolated 26 colonies of Mycobacterium spp. and 6 species have been identified, all of them are environmental Mycobacterium fortuitum, M. confluentis, M. elephantis, M. novocastrense, M. sphagni and M. arupense; 7 isolates, however, remained without characterization for the specie. M. fortuitum is an important opportunistic pathogen in public health, without, however, evidence of being transmitted by food; M. novocastrense, M. arupense and M. elephantis species have also been considered potentially pathogenic to humans. The results suggest that the frequency and/or contamination load of M. bovis in meia cura Minas cheese is (are) low (s). We have to consider, however, that this result may have been influenced by the absence of an analytical method capable of identifying the agent in the food matrix in which a low load of microorganism is expected, accompanied by high load of contaminants. Also suggest the need to evaluate the possible importance of foodborne non-tuberculous mycobacteria, especially for immunocompromised individuals.

Made available in DSpace on 2014-07-29T15:22:58Z (GMT). No. of bitstreams: 1 dissertacao flavio pereira.pdf: 1203680 bytes, checksum: bcaf62e9eb5545119172fb4728bdaaee (MD5) Previous issue date: 2010-02-26 / Raw milk is a natural culture medium for the microbial growth and easy deterioration, because of that, thermal processing is needed as soon as possible. A new technology has been used to improve the food quality, active packaging, such as antimicrobial films, wich the intention to control the microbial growth in the products. The aim of this study was to evaluate the UHT milk quality produced in Goiás State, isolation and identification of Bacillus sporothermodurans, beside create a antimicrobial film for inhibition of this microrganism high heat resistance spore former. The isolation of psychrotrophic in raw milk was used PCA with incubation at 21 °C for 72h. To evaluate the UHT milk has been doing research on aerobic mesophilic in PCA with incubation at 30 °C for 72h and to thermoresistents microrganisms was in ABHI with incubation at 35 °C for 48h. The identification of isolates was performed according to convetional methods of biochemistry identification. From raw milk, the psychrotrophic microrganisms most isolates were Staphylococcus sp. and Corynebacterium sp., both with 21,5% from 70 isolates, verifying a higher contauge of Gram-positive microrganisms (82,85%). Considering the 157 UHT isolates, just 31 were identified as Gram-positive microrganisms through biochemical tests, only 2 correponded to B. sporothermodurans, although there are differences from the tests and their results between various authors, requiring more accurate identification using molecular biology. The antimicrobial film maded of cellulose acetate with addition of nisin to inhibit the development of B. sporothermodurans, the results indicated action of nisin in inhibition this microrganism, however, when incorporated into the film, did not show the expected results in both tests, solid and liquid medium, although the specific rate (&#956;) growth of the microrganism was changing iin the lowest concentration tested. There is a need to conduct testing in foods, mainly in dairy products which have been reported contamination by B. sporothermodurans. / O leite por ser um alimento rico, constituí-se em um meio natural para o desenvolvimento de vários microrganismos, principalmente bactérias; desta forma, muito perecível. É necessário tratamento térmico o mais rápido possível, a fim de manter a qualidade. Uma nova tecnologia vem sendo utilizada na melhoria dos alimentos, denominada de embalagem ativa, como os filmes antimicrobianos, na intenção de controlar o desenvolvimento microbiano no produto. O objetivo deste trabalho foi de avaliar a qualidade de leite UAT produzido no Estado de Goiás, assim como isolar e identificar microrganismos psicrotróficos de leite cru e microrganismos de leite UAT, sendo que para este direcionado para identificação de Bacillus sporothermodurans, bem como criação de filme antimicrobiano para inibição deste microrganismo produtor de esporos altamente resistentes. Para o isolamento de psicrotróficos em leite cru, foi utilizado PCA e incubação à 21°C por 72h. Para avaliação de qualidade do leite UAT, foi feito a pesquisa de aeróbios mesófilos em PCA com incubação à 30°C por 72h e o isolamento de microrganismos termorresistentes foi realizado em ABHI com incubação a 35°C por 48h. A identificação dos isolados foi realizada conforme metodologias clássicas de identificação bioquímica. Segundo esta identificação, os microrganismos psicrotróficos mais isolado foram Staphylococcus sp. e Corynebacterium sp. com 21,5% cada, dos 70 isolados, verificando uma percentagem maior de microrganismos Gram-positivos (82,85%). Considerando os isolados de leite UHT, dos 157, apenas 31 foram identificados como sendo bastonetes Gram-positivos que submetidos a testes bioquímicos, apenas 2 corresponderam ao B. sporothermodurans seguindo-se os resultados conforme registrado, apesar de haver divergências quanto a alguns testes e seus resultados, necessitando de identificação mais apurada utilizando-se de biologia molecular. Quanto ao filme antimicrobiano a base polimérica utilizada foi o acetato de celulose com incorporação de nisina, a fim de inibir o desenvolvimento do B. sporothermodurans com o intuito de verificar a ação deste peptídeo antimicrobiano frente a este microrganismo. Os resultados obtidos indicaram ação da nisina na inibição do microrganismo estudado, no entanto, quando incorporado ao filme, não apresentou o resultado esperado, tanto em testes em meio sólido como em meio líquido, apesar de ter alterando a velocidade específica (&#956;) de crescimento do microrganismo na concentração mais baixa testada. Há a necessidade de realização de teste em alimentos, especialmente em leite e derivados em que foram relatados a contaminação pelo B. sporothermodurans