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Isolation of PCR-ready DNA from probiotic products for baby nutrition
Mantlová, Gabriela ; Havlíková, Šárka (referee) ; Španová, Alena (advisor)
The aim of thesis is focused on isolation of DNA in quality for polymerase chain reaction (PCR) and the identification of probiotic bacteria. From six probiotic supplements for children were isolated PCR-ready DNAs using magnetic carriers P(HEMA-co-GMA). Isolated DNA was amplified by genus-specific and species-specific primers. DNAs of Lactobacillus, Bifidobacterium and Streptococcus genera were identified as: L. acidophilus, L. rhamnosus, L. casei, B. bifidum, B. longum ssp. longum, B. breve, B. longum ssp infantis, B. animalis and S. thermophilus. The identification corresponded with the data declared by the producers.
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Isolation of PCR-ready DNA from dairy products for baby nutrition
Mantlová, Jana ; Eva, Kvasničková (referee) ; Španová, Alena (advisor)
The work was focused on isolation of PCR-ready DNA and the identification of probiotic lactic acid bacteria that were isolated from five milk product for infant nutrition. DNA was isolated from crude cell-lysates of the products by magnetic P(HEMA-co-GMA) microspheres. DNAs isolated from crude cell lysates of control strains using phenol extraction method were used as positive controls. Using PCRs DNA of genera Bifidobacterium and species B. animalis, B. bifidum, B. breve, B. infantis, B. longum and Streptococcus thermophilus species were identified in products. The results obtained are consistent with the data declared by the manufacturers.
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Selective isolation of the genus Bifidobacterium bacteria from foods
Mizerovská, Lucie ; Šárka, Havlíková (referee) ; Rittich, Bohuslav (advisor)
Probiotic lactic acid bacteria (LAB) are very often used in food procesing industry, such as milk products, cheese and fermentsd salami production in nova days. In diploma thesis were tested symbiotic food supplements from different producers. Bacterial DNA was isolated from crude cell lysates of six food suplements by magnetic particles P(HEMA-co-GMA). PCR-ready DNAs were isolated. from all products The detection of Bifidobacterium bacteria identified by PCR was in agreement with those declared by the manufacturers. Magnetic particles with immobilized antibodies against Bifidobacterium were used in the next part of thesis. These particles were used for the isolation of target cells from two products with cell identification by genus specific PCR.
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PCR identification of nonpathogenic bacteria strains in cheeses
Jurečková, Nela ; Doušková, Dagmar (referee) ; Španová, Alena (advisor)
Different species of genus Bifidobacterium are part of human and animal intestinal flora. These bacteria have benefit effects and therefore they are used in foods and pharmaceutical products as probiotics. Cheese is now suitable as a probiotic matrix except yoghurts and fermentated milks. This diploma thesis was focused on optimalization of DNA isolation from bacteria of genus Bifidobacterium. Magnetic microparticles (P(HEMA-co¬-GMA)) were used for DNA isolation in presence of 8% polyethyleneglycol PEG 6000 and 5 M sodium chloride. Phenol extraction weas also used as an isolation method. Isolated DNA was used for amplification in domain, genus and species specific PCRs. Optimized method was tested for detection of bacteria of genus Bifidobacterium in experimentaly prepared probiotic cheeses. These cheeses contained potential probiotic bacteria from Laktoflóra collection. Bacteria were identified into species using species specific PCR. Species Bifidobacterium animalis was identified in all samples of probiotic cheeses.
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Probiotic bacteria in pharmaceutical products
Suranová, Mária ; Vránová, Dana (referee) ; Veselá, Mária (advisor)
Probiotics are live microorganisms, that when administered in adequate amount confer a health benefit on the host. They are used as part of specific milk products and either as dietary supplements in form of tablets, capsules, coated pills and sirups. The most famous probiotic microorganisms are bacteria of Lactobacillus and Bifidobacterium genus, that belong in group of lactic acid bacteria. For the assessment of probiotic preparations is used broad range of methods for the microbial analysis of foods. These methods can be separated on quantitative, that enumerate amount of microorganisms in product and qualitative, that are capable detect presence or absence of certain microorganisms. Both types of the methods have large importance in control of the efficiency probiotic preparations.
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Identification and characterization of selected species of lactic acid bacteria
Trojánek, Zdeněk ; Špano,, Miroslav (referee) ; Rittich, Bohuslav (advisor)
Among the important representative of lactic acid bacteria are gram-positive bacteria of the genus Bifidobacterium. They are constituent part of ordinary intestinal flora, which help keep the balance and prevent colonization of intestinal by pathogens. Bifidobacteria are used in various dairy products for their prebiotic effect. Molecular biological methods are used for rapid and accurate identification of bacteria of genus Bifidobacterium. In our case, we have focused on DNA isolation and application of the polymerase chain reaction (PCR) for identification of bacteria of genus Bifidobacterium. Statistical evaluation of fingerprints obtained by rep-PCR with (GTG)5 primer was carrid out.
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Identification of selected probiotic bacteria in food additives
Bubeníková, Lucia ; Španová, Alena (referee) ; Rittich, Bohuslav (advisor)
Bifidobacteria and lactobacilli are gram-positive bacteria which belong to the lactic acid bacteria group. They are ordinary constituents of gastrointestinal microflora, play a pivotal role in human nutrition and health, prevent pathogen colonization and maintain normal mucosal immunity. In present days they are used in production of functional food products and pharmaceutical additives. Polymerase chain reaction (PCR) is used to the detection and identification of bacteria which belong to the genus of Bifidobacterium and Lactobacillus. Specific primers for each genus are used in PCR reactions for in vitro amplification of definite part of DNA. In this work, total DNA was isolated from three additives (Pangamin Bifi plus, Biopron Junior and Probiodom) by the method of phenol extraction and amplified using genus-specific PCR primers giving amplicons of 523 bp (Bifidobacterium) and 250 bp (Lactobacillus). Occurrence of bacteria of both genera was proved in all of the three tested probiotic products.
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