Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-11-18T16:24:01Z No. of bitstreams: 2 jakline_magela_santos.pdf: 282068 bytes, checksum: 45afba7935a8b53d9cee205b7c963573 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-11-18T16:24:24Z (GMT) No. of bitstreams: 2 jakline_magela_santos.pdf: 282068 bytes, checksum: 45afba7935a8b53d9cee205b7c963573 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Made available in DSpace on 2015-11-18T16:24:24Z (GMT). No. of bitstreams: 2 jakline_magela_santos.pdf: 282068 bytes, checksum: 45afba7935a8b53d9cee205b7c963573 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Previous issue date: 2010 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Funda??o de Amparo ? Pesquisa do estado de Minas Gerais (FAPEMIG) / Objetivou-se verificar as caracter?sticas f?sico-qu?micas e microbiol?gicas do leite cru proveniente de tanques da regi?o de Curvelo, Minas Gerais, e avaliar o crescimento de microrganismos psicrotr?ficos no leite cru refrigerado nas temperaturas de 4, 7 e 10?C. Constatou-se que 9,09% dos tanques estiveram em desacordo com a legisla??o quanto ao ?ndice criosc?pico e 22,72% quanto ? acidez dornic. Os testes f?sico-qu?micos n?o detectaram presen?a de ?gua oxigenada, formol, ou subst?ncias alcalinas em 100% das amostras analisadas. Todas as amostras dos tanques apresentaram teor de gordura acima de 3%. Para contagem de mes?filos 81,81% dos tanques avaliados mantiveram-se abaixo de 7,5x105 UFC/mL e 90,91% dos tanques avaliados apresentaram contagens abaixo da regulamenta??o para a contagem de c?lulas som?ticas que ? de 7,5x105 c?lulas/mL. Na contagem de psicrotr?ficos 72,72% dos tanques apresentaram contagens superiores em mais de 10% ? contagem de mes?filos. Os tempos de redu??o do azul de metileno apresentaram-se acima de 90 minutos em 90,91% dos tanques. O crescimento de psicrotr?ficos no leite cru refrigerado foi mais lento a 4?C do que nas temperaturas de 7 e 10?C (p<0,05) em todos os tempos avaliados. Nas temperaturas de 7 e 10?C, n?o houve diferen?a (p>0,05) no crescimento em todos os tempos avaliados. O crescimento m?dio de psicrotr?ficos a 10?C foi 4 vezes maior que a 4?C, enquanto que a 7?C foi 3,3 vezes maior que o crescimento a 4?C. Conclui-se que o leite proveniente dos tanques est? de acordo com os par?metros previstos na legisla??o, no entanto, a qualidade do leite est? comprometida devido ?s elevadas contagens de psicrotr?ficos. Quanto o crescimento de psicrotr?ficos um tempo m?ximo de estocagem deve ser utilizado como par?metros de identidade e qualidade para leite cru refrigerado / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Zootecnia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2010. / ABSTRACT This study assessed the physicochemical and microbiological characteristics of raw milk came from tanks from Curvelo region, Minas Gerais, Brazil, and evaluating the growth of psychrotrophic in raw milk refrigerated at temperatures of 4.7 and 10? C. It was found that 9.09% of the tanks were not in compliance with the legislation regarding the cryoscopic index and 22.72% for the Dornic acidity. The physical-chemical tests found no presence of hydrogen peroxide, formaldehyde, or alkaline substances in 100% of the samples analysed. All samples from the tanks had fat above 3%. The mesophilic counting of 81.81% of the tanks assessed remained below 7.5x105 CFU/mL and 90.91% of the tanks evaluated presented under regulation scores for somatic cell count that is 7.5x105 cells/mL. For the psychrotrophic counting, 72.72% of the tanks had higher scores in more than 10% of mesophilic counting. The time of reduction of methylene blue showed up after 90 minutes in 90.91% of the tanks. The growth of psychrotrophic in refrigerated raw milk was slower at 4?C than at temperatures of 7 and 10?C (p <0.05) at all times evaluated. At temperatures of 7 and 10?C, there has been no difference (p> 0.05) on growth at all times evaluated. The average growth of psychrotrophic at 10?C was four times higher than at 4?C, while at 7?C it was 3.3 times greater than the growth at 4?C. It is concluded that milk from the tanks complies with the parameters set in legislation; however, the quality of milk is hampered by the high psychrotrophic population. Regarding the growth of psychrotrophic a maximum time of storage should be used as parameters of identity and quality of refrigerated raw milk.

Made available in DSpace on 2014-12-17T15:01:18Z (GMT). No. of bitstreams: 1 PriscillaDLS.pdf: 447651 bytes, checksum: a15751ca9e9de37e7c6b4041718d3642 (MD5) Previous issue date: 2005-12-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Extended storage of refrigerated milk can lead to reduced quality of raw and processed milk, which is a consequence of the growth and metabolic activities of psychrotrophic bacteria, able to grow under 7oC or lower temperatures. Although most of these microorganisms are destroyed by heat treatment, some have the potential to produce termoresistant proteolytic and lipolytic enzymes that can survive even UHT processing and reduce the processed products quality. Recently, the IN 51 determineds that milk should be refrigerated and stored at the farm what increased the importance of this group of microorganisms. In this work, psychrotrophic bacteria were isolated from 20 communitarian bulk tanks and 23 individual bulk tanks from dairy farms located at Zona da Mata region of Minas Gerais State and from southeastern Rio de Janeiro. Selected milk dilutions were plated on standard agar and after incubation for 10 days at 7oC, five colonies were isolated, firstly using nutrient agar and after using McConkey agar for 24 hours at 21oC. The isolates were identified by morphology, Gram stain method, catalase production, fermentative/oxidative metabolism and by API 20E, API 20NE, API Staph, API Coryne or API 50 CH (BioMerieux). In order to ensure reproductibility, API was repeated for 50% of the isolates. Species identification was considered when APILAB indexes reached 75% or higher. 309 strains were isolated, 250 Gram negative and 59 Gram positive. 250 Gram negative isolates were identified as: Acinetobacter spp. (39), Aeromonas spp. (07), A. Hydrophila (16), A. sobria (1), A. caviae (1), Alcaligenes feacalis (1), Burkholderia cepacia (12), Chryseomonas luteola (3), Enterobacter sp. (1), Ewingella americana(6), Hafnia alvei (7), Klebsiella sp. (1), Klebsiella oxytoca (10), Yersinia spp. (2), Methylobacterium mesophilicum (1), Moraxella spp. (4), Pantoea spp. (16), Pasteurella sp. (1), Pseudomonas spp. (10), P. fluorescens (94), P. putida (3), Serratia spp. (3), Sphigomonas paucomobilis (1). Five isolates kept unidentified. Pseudomonas was the predominant bacteria found (43%) and P. fluorescens the predominant species (37.6%), in accordance with previous reports. Qualitative analysis of proteolytic and lipolytic activity was based on halo formation using caseinate agar and tributirina agar during 72 hours at 21oC and during 10 days at 4?C, 10oC and 7?C. Among 250 Gram negative bacteria found, 104 were identified as Pseudomonas spp. and 60,57% of this group showed proteolytic and lipolytic acitivities over all four studied temperatures. 20% of Acinetobacter, Aeromonas, Alcaligenes, Burkholderia, Chryseomonas, Methylobacterium, Moraxella presented only lipolytic activity. Some isolates presented enzymatic activity in one or more studied temperatures. Among Gram positive bacteria, 30.51% were proteolytic and lipolytic at 10oC, 8.47% were proteolytic at 7oC, 10oC, and 21oC, 8.47% were proteolytic at all studied temperatures (4oC, 7oC, 10oC and 21oC) and 3.38% were proteolytic only at 21oC. At 4oC, only one isolate showed proteolytic activity and six isolates were lipolytic. In relation to Gram negative microorganisms, 4% were proteolytic and lipolytic at 7oC, 10oC and 21oC, 10% were proteolytic at 10oC and 4.4% were lipolytic at 4oC, 7oC, 10oC and 21oC, while 6.4% of all isolates were proteolytic and lipolytic at 10oC and 21oC as well as lipolytic at 4oC and 7oC. These findings are in accordance with previous researches that pointed out Pseudomonas as the predominant psycrotrophic flora in stored refrigerated raw milk / O processo de conserva??o do leite cru em temperaturas de refrigera??o por per?odos prolongados pode acarretar perda de qualidade do leite e, conseq?entemente, de seus produtos derivados, fato associado ao crescimento e ? atividade enzim?tica de bact?rias psicrotr?ficas, as quais podem se desenvolver em temperaturas abaixo de 7?C. Estes microrganismos geralmente s?o eliminados durante o tratamento t?rmico do leite, por?m suas enzimas proteol?ticas e lipol?ticas s?o termoest?veis, podendo resistir at? mesmo ao tratamento UAT e permanecerem ativas causando perda de qualidade dos produtos processados. Recentemente a INSTRU??O NORMATIVA 51, estabelece que o leite deve ser refrigerado e armazenado na propriedade rural, o que aumentou a import?ncia desse grupo de bact?rias. Neste trabalho, foram isoladas e identificadas bact?rias contaminantes de leite cru refrigerado proveniente de 20 tanques comunit?rios e 23 tanques individuais de propriedades leiteiras da regi?o da Zona da Mata do Estado de Minas Gerais e sudeste do Rio de Janeiro. Dilui??es selecionadas foram plaqueadas em agar padr?o e ap?s incuba??o a 7?C/10 dias, 5 col?nias foram isoladas em placas contendo agar nutriente, com incuba??o a 21?C/24 h. Os isolados foram identificados de acordo com a morfologia, colora??o de Gram, produ??o de catalase, oxidase, metabolismo fermentativo/oxidativo, crescimento em meio Mac Conkey, rea??o de KOH 3%, hidr?lise da arginina, motilidade, produ??o de H2S e indol em meio SIM e presen?a de esporos, e pelo Sistema API 20E, API 20NE, API Staph, API Coryne ou API 50 CH (BioM?rieux). O sistema API foi repetido para 50% dos isolados para garantir a reprodutibilidade dos resultados. A identifica??o em n?vel de esp?cie foi considerada quando o programa APILAB indicou identidade &#8805; 75%. Foram obtidos 309 isolados, sendo 250 Gram negativos e 59 Gram positivos. Os 250 isolados Gram negativos obtidos foram identificados como: Acinetobacter spp. (39), Aeromonas spp. (07), A. Hydrophila (16), A. sobria (1), A. caviae (1), Alcaligenes feacalis (1), Burkholderia cepacia (12), Chryseomonas luteola (3), Enterobacter sp. (1), Ewingella americana(6), Hafnia alvei (7), Klebsiella sp. (1), Klebsiella oxytoca (10), Yersinia spp. (2), Methylobacterium mesophilicum (1), Moraxella spp. (4), Pantoea spp. (16), Pasteurella sp. (1), Pseudomonas spp. (10), P. fluorescens (94), P. putida (3), Serratia spp. (3), Sphigomonas paucomobilis (1). Cinco isolados n?o puderam ser identificados por meio dos testes utilizados. Pseudomonas foi o g?nero mais isolado (43%) sendo Pseudomonas fluorescens a esp?cie predominante (37,6%), o que corrobora dados da literatura. A produ??o de enzimas proteol?ticas e lipol?ticas foi avaliada qualitativamente pela presen?a de halo em torno da col?nia em agar caseinato e agar tributirina durante 72 horas a 21?C e 10 dias a 4?C, 7?C e 10oC. Do total de 250 Gram negativos, 104 foram identificados como Pseudomonas spp., das quais 60,57% apresentaram atividade proteol?tica e lipol?tica nas quatro temperaturas estudadas. 20% das esp?cies pertencentes aos g?neros: Acinetobacter, Aeromonas, Alcaligenes, Burkholderia, Chryseomonas, Methylobacterium, Moraxella, demonstraram apenas atividade lipol?tica nas quatro condi??es estudadas. Dos 59 isolados Gram-positivas obtidos, foram identificados os seguintes g?neros: Kurthia spp (7), Levedura (1), B. stearothermophilus (1), Bacillus coagulans (1), Bacillus lentus (1), Brevibacterium spp (1), Cellum/Microbacterium (6), Staphylococcus spp (3). Trinta e sete isolados n?o puderam ser identificados por meio dos testes utilizados. Alguns isolados apresentaram atividade enzim?tica em uma ou mais das quatro temperaturas estudadas: Gram-positivos- 30,51% foram proteol?ticas a 7?, 10 e 21?C e lipol?tica a 10?C; 8,47% foram proteol?ticos a 7?, 10? e 21?C; 8,47% foram lipol?tica nas quatro temperaturas estudadas; 3,38% proteol?ticos s? a 21?C e somente um isolado tiveram atividade proteol?tica a 4?C e seis atividade lipol?tica a esta mesma temperatura. Gram negativos - 4% foram proteol?tico e lipol?tico a 7?, 10? e 21?C; 10% proteol?ticas a 10?C; 4,4% lipolitico a 4?, 7? e 21?C; 6,4% foram proteol?tico e lipol?tico a 10? e 21?C e tamb?m lipol?tico a 4? e 7?C. Estes resultados s?o consistentes com os de outras pesquisas, em que tamb?m foram constatados que esp?cies do g?nero Pseudomonas representam a microbiota psicrotr?fica deterioradora mais freq?ente do leite refrigerado