Rapid Identification of Streptococcus thermophilus by Primer-specific PCR Amplification Based on its lacZ Gene

Lick, Sonja; Keller, Martina; Bockelmann, Wilhelm; Heller, Jochem

doi:10.1016/s0723-2020(96)80012-9

Cited by 68 publications

(49 citation statements)

References 17 publications

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“…Resultados semelhantes foram encontrados por Feitosa et al (2003), que, ao estudarem microrganismos indicadores de higiene em queijos produzidos no Rio Grande do Norte, constataram a presença de coliformes termotolerantes em 36,4% das amostras estudadas, 3 a 7NMP/g, dentro do limite descrito na legislação. Duarte et al (2005) Estes resultados se assemelham aos de vários relatos sobre a análise da microbiota de queijos de origem artesanal em várias partes do mundo, como o queijo São Jorge, produzido em Portugal (Kongo et al, 2007), queijo Raschera, proveniente da Itália (Dolci et al, 2008b), Darfiyeh, encontrado na Líbia (Serhan et al, 2009), queijos espanhóis artesanais de leite de cabra (Martín-Platero et al, 2009), queijo Fontina, originado da Itália (Giannino et al, 2009), e queijo Istriano, presente na Croácia (Fuka et al, 2010). Esses autores sempre relacionaram as espécies do gênero enterococos aos aspectos organolépticos dos produtos artesanais, ressaltando a participação na maturação dos queijos.…”

Section: Resultsunclassified

Avaliação da microbiota bacteriana do queijo de coalho artesanal produzido na região Agreste do estado de Pernambuco

Silva

Bismara

Moura

et al. 2012

Arq. Bras. Med. Vet. Zootec.

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RESUMOO objetivo desta pesquisa foi avaliar a qualidade microbiológica e o perfil ácido-láctico do queijo de coalho artesanal. Todas as amostras de queijo apresentaram coliformes totais, termotolerantes e presença de Escherichia coli, porém com os valores dentro dos padrões estabelecidos pela legislação vigente no país. O perfil ácido-láctico estudado mostrou uma microbiota heterogênea, constituída por lactobacilos, lactococos, estreptococos e enterococos, confirmadas as espécies Enterococcus faecalis, Enterococcus faecium, Streptococcus thermophilus e Lactococcus lactis.Palavras-chave: queijo de coalho artesanal, reação em cadeia da polimerase, bactérias ácido-lácticas ABSTRACTThe aim of this study was to evaluate the microbiological quality and lactic-acid profile of artisanal coalho cheese. All cheese samples analyzed showed total coliforms, were thermotolerant, and had Escherichia coli, but all the values were within the standards established by current legislation in the country, and could be considered a food fit for human consumption. The cheese showed a heterogeneous microbiota, being constituted of all tested genus, such as lactobacilli, lactococcus, streptococcus and enterococcus, and confirmed the species: Enterococcus faecalis, Enterococcus faecium, Streptococcus thermophilus and Lactococcus lactis. Keywords: artisanal coalho cheese, polymerase chain reaction, lactic-acid bacteria INTRODUÇÃO A produção de queijo de coalho representa uma atividade bastante significativa para a economia regional, visto que em determinadas localidades é a principal fonte de renda e sobrevivência da população (Almeida et al., 2010). No Nordeste do Brasil a maior parte da produção de queijo de coalho é realizada em pequenas e médias queijarias, as quais movimentam, mensalmente, algo em torno de 10 milhões de reais, o que torna essa atividade importante tanto no âmbito social quanto no econômico (Perry, 2004).Recebido em 1 de julho de 2011 Aceito em 7 de agosto de 2012 *Autor para correspondência (corresponding author) E-mail: mtcvsoares@yahoo.com.br De acordo com Almeida et al. (2010), o queijo de coalho é uma representação genuína da tradição e cultura do estado de Pernambuco, onde está sendo desenvolvido um trabalho de certificação desse queijo visando a uma identidade própria que irá diferenciá-lo dos demais produzidos na região Nordeste.As propriedades organolépticas típicas e o aroma particular dos queijos obtidos com leite cru, tais como o queijo de coalho artesanal, estão associados com atributos do próprio leite, relacionados à raça e ao tipo de nutrição das vacas, ao processo de fabricação básico do queijo tradicional e à microbiota natural autóctone,

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Section: Resultsunclassified

Avaliação da microbiota bacteriana do queijo de coalho artesanal produzido na região Agreste do estado de Pernambuco

Silva

Bismara

Moura

et al. 2012

Arq. Bras. Med. Vet. Zootec.

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“…Twenty six cultures were discarded on the basis of biochemical tests and remaining 76 were subjected to molecular characterization for definite confirmation. Amplification of species specific lacZ gene of S. thermophilus was carried out using universal primer pairs (ST 1 /ST 2 ) devised by Lick et al (1996). On the basis of species specific PCR, out of 76 biochemically identified isolates 62 were confirmed as S. thermophilus (26 from raw milk, 18 from dahi, 14 from lassi and 4 from yoghurt).…”

Section: Discussionmentioning

confidence: 99%

“…These tests include catalase test; growth at 10°C, 45°C, growth at pH 9.6; resistance to heating at 65°C for 30 min, growth in 2 % and 4 % NaCl, 0.04 % K-tellurite and in 0.1 % methylene blue; arginine hydrolysis; esculin hydrolysis test and carbohydrate fermentation (glucose, fructose, sucrose, mannose, mannitol, sorbitol, cellibiose, arabinose, lactose, mellibiose, galactose, xylose, rhamnose, maltose and inulin). All the biochemically identified S. thermophilus isolates were subjected to molecular characterization for definite confirmation of species using PCR by targeting strongly conserved sequences of S. thermophilus lacZ gene (Lick et al 1996). The primers were custom synthesized from Integrated DNA technologies, U.S.A. (Table 1).…”

Section: Methodsmentioning

confidence: 99%

Production of β-galactosidase from streptococcus thermophilus for galactooligosaccharides synthesis

et al. 2014

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Efficiency of different methods for disruption of Streptococcus thermophilus cells, isolated from different dairy products, to release β-galactosidase and synthesis of GOS by extracted enzyme using whey supplemented with different concentrations of lactose as a substrate was studied. Unlike most other studies on GOS synthesis which used only one method of cell disruption and only few microbial strains, we compared five different cell disruption methods and used 30 strains of S. thermophilus in order to find out the most effective method and efficient strain for production of β-galactosidase. Appreciable amount of GOS (53.45 gL −1 ) was synthesized at a lactose concentration of 30 %, using enzyme (10 U mL −1 of reaction medium), extracted from S. thermophilus within a very short incubation time of 5 h at a temperature of 40°C and pH 6.8. S. thermophilus is heavily employed in the preparation of fermented dairy products but this study extends the use of this organism for the production of GOS, a potential prebiotic.

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“…The phenotypically characterized isolates were further genetically identified by species specific polymerase chain reaction (PCR) based on LacZ gene as previously described by Lick et al (1996). S. thermophilus isolates were grown in M17 broth, and genomic DNA was extracted as previously described (Pospiech and Neumann 1995).…”

Section: Molecular Identificationmentioning

confidence: 99%

“…S. thermophilus isolates were grown in M17 broth, and genomic DNA was extracted as previously described (Pospiech and Neumann 1995). PCR was performed for the identification of isolates on an Eppendorf Mastercycler (Hamburg, Germany) according to earlier published literature (Lick et al 1996). Amplification was verified by electrophoresis on 1.5% (w/v) agarose gel in 1× Tris-acetate-EDTA buffer.…”

Section: Molecular Identificationmentioning

confidence: 99%

Bioprospecting of strains of Streptococcus thermophilus from Indian fermented milk products for folate production

Iyer

Tomar

Mohanty

et al. 2011

Dairy Science & Technol.

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This study deals with the bio-prospecting of folate producing strains of Streptococcus thermophilus isolated from milk and different fermented milk products of Indian origin. From a total of 500 randomly selected colonies isolated from 209 different samples, 117 isolates were identified as S. thermophilus by classical biochemical and molecular characterization. Frequency of incidence of S. thermophilus in the different samples of milk and milk products was variable with the highest in the dahi followed by yogurt and lassi and a very low incidence in case of milk and cheese. On screening for folate using a microbiological assay with a trienzyme extraction, about 15% of strains was found to produce folate in the range of 40-50 μg·L −1 , 35% in the range of 20-30 μg·L −1 , and the remainder in the range of 4-16 μg·L −1 . Comparative analysis of the random amplification of polymorphic DNA PCR fingerprint profiles was used to characterize interspecific diversity of the ten highest folate producers. The LacZ gene of two of the highest folate producing isolates were sequenced and submitted to GenBank under following accession numbers of FJ161697 and FJ161698.

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Rapid Identification of Streptococcus thermophilus by Primer-specific PCR Amplification Based on its lacZ Gene

Cited by 68 publications

References 17 publications

Avaliação da microbiota bacteriana do queijo de coalho artesanal produzido na região Agreste do estado de Pernambuco

Avaliação da microbiota bacteriana do queijo de coalho artesanal produzido na região Agreste do estado de Pernambuco

Production of β-galactosidase from streptococcus thermophilus for galactooligosaccharides synthesis

Bioprospecting of strains of Streptococcus thermophilus from Indian fermented milk products for folate production

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