Typing of bacteriophages by randomly amplified polymorphic DNA (RAPD)-PCR to assess genetic diversity

Gutiérrez, Diana; Martín‐Platero, Antonio M.; Rodrı́guez, Ana; Martínez‐Bueno, Manuel; Garcı́a, Pilar; Martı́nez, Beatriz

doi:10.1111/j.1574-6968.2011.02342.x

Cited by 66 publications

(41 citation statements)

References 33 publications

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“…Diana Gutiérrez et al (2011) have suggested that cocktails composed of phages with different genetic characteristics are more effective in phage therapy. They have shown that band patterns obtained by RAPD-PCR are an easy and reproducible method for bacteriophage typing (30).…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Listeria monocytogenes Bacteriophages from Environmental Sources in Kars-Turkey

Mutlu¹

2019

PVJ

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In this study, 45 samples were collected from some commercial businesses and environmental focus located in Kars center. Dirty areas, the area in contact with the sewage and poultry environmental sources were preferred while collecting samples. Again, 3 different silage samples and two samples were suspicious of Listeria were evaluated that brought to Kafkas University, Faculty of Veterinary Microbiology Laboratory. Isolation of lytic phages and lysogenic phages with mitomycin-C from total of 50 samples was performed. From 1 out of 12 of these samples lysogenic, from the remaining 11 samples lytic phage isolation was made. Isolation was confirmed by plaque assay. RAPD-PCR method was used for genotyping. The agarose gel images of 3 phages were observed to have been determined as previous study vB_SepiS-philPLA7, vB_SauS-philPLA35 and ΦH5. When band profiles belong to remaining 9 phages were compared, band profiles of 3 phages and the other 5 phages band profiles evaluated to be the same with each other. This study is thought to be precursors for purposes such as the decontamination of silage will be produced in the region and animal food, since Listeria infections are important for animal and human health and cause considerable economic losses.To Cite This Article: Mutlu N and Şahin M, 2019. Isolation and characterization of Listeria monocytogenes bacteriophages from environmental sources in Kars-Turkey. Pak Vet J, 39(1): 91-95. http://dx.

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Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Listeria monocytogenes Bacteriophages from Environmental Sources in Kars-Turkey

Mutlu¹

2019

PVJ

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“…DNA was extracted from colonies as described above. Primers OPL5 (5'-ACGCAGGCAC-3'), RAPD5 (5'-AACGCGCAAC-3'), P1 (5'-CCGCAGCCAA-3') were used according to Gutiérrez et al (2011). RAPD-PCR reactions were performed with the following cycling program: four cycles at 94 °C for 45 s, 30 °C for 120 s and 72 °C for 60 s; 26 cycles at 94 °C for 5 s, 36 °C for 30 s and 72 °C for 30 s and a final step of 10 min at 75 °C.…”

Section: Genetic Fingerprinting By Rapd-pcrmentioning

confidence: 99%

Biodiversity of Bacteriocin-Producing Lactic Acid Bacteria from Mexican Regional Cheeses and their Contribution to Milk Fermentation

et al. 2016

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The aim of this work was to examine the biodiversity of bacteriocin-producing lactic acid bacteria from homemade cheeses produced in Veracruz (México) and assess their contribution as adjunct cultures in dairy products. Ninety-three presumptive bacteriocinogenic strains were detected by direct antagonism assays and twenty-nine out of them were active against Enterococcus faecalis NRRL-B537, Listeria innocua 062 AST, or Listeria monocytogenes ATCC19115 by the well diffusion test using cell-free supernatants, adjusted to pH 6.0 to exclude inhibition by organic acids. Positive isolates were identified by partial sequencing of the 16s rDNA as Pediococcus acidilactici (4 isolates), Enterococcus faecium (17 isolates), Lactobacillus plantarum (6 isolates) and Lactobacillus fermentum (2 isolates). RAPD-PCR discriminated 7 groups with a 50% similarity and revealed the presence of the same isolates. The coding genes for the synthesis of plantaricin EF, plantaricin JK, plantaricin N, plantaricin NC8 and the inducing peptide plantaricin A were detected by PCR in L. plantarum. Similarly, enterocin P and pediocin PA-1 genes were amplified from Enterococcus and Pediococcus genomic DNA, respectively. Overall, co-culturing of bacteriocin producing Lactobacillus and Pediococcus strains with the dairy starter Lactococcus lactis IPLA947 did not interfere with milk acidification. Lactose consumption, acidification rate and production of lactic acid were unchanged. Nonetheless, higher levels of acetic acid, ethanol and succinic acid were detected depending on the strain. Our results demonstrate the diversity of bacteriocinogenic species in homemade Mexican cheeses which may be used as adjunct cultures to enhancing safety of this wellappreciated cheese while providing a richer range of metabolites.

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“…Phages were induced by UV light and purified as described in Materials and Methods. At the end of the treatment with UV, the phage count was estimated by epifluorescence at 10 10 phage/ml, 1 log unit higher than that reported to be needed for optimal reproducibility (29). The 15 phage DNAs were subjected to two RAPD-PCRs, one using only the random primer M13 and, to increase the sensitivity of the profiles (22), a second one with primers M13 and Lys20_fw.…”

Section: Pcr Screening and Phage Characterization By Lys Prophage Seqmentioning

confidence: 99%

Development of a New Method for Detection and Identification of Oenococcus oeni Bacteriophages Based on Endolysin Gene Sequence and Randomly Amplified Polymorphic DNA

Doria

Napoli

Costantini

et al. 2013

Appl Environ Microbiol

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M alolactic fermentation (MLF) is a secondary fermentation that occurs in wine after the alcoholic fermentation. This fermentation can take place spontaneously through the action of the population of lactic acid bacteria (LAB) present in the wine or by the use of commercial starters, mainly belonging to the species Oenococcus oeni, the principal LAB species responsible for MLF in wine (1, 2).As in other fermented foods, like dairy products, in which bacteriophages represent a potential risk for the fermentative process (3, 4), bacteriophages have also been reported to be a possible cause of unsuccessful MLF in wine (5, 6). Therefore, preparation of commercial starters that take into account the different sensitivities of O. oeni strains to different phages has been proposed (5, 7). However, under laboratory conditions, O. oeni has a very low growth rate, and its poorly visible colonies are hardly produced in agar medium, hampering the accurate detection of O. oeni phages, which has contributed to the low number of studies conducted until now on this topic. For instance, while PCR-based approaches to detect bacteriophages have been used in bacteria of the genera Lactobacillus and Lactococcus present in dairy products (8-11), such methods have never been applied to detect phages infecting O. oeni. In the present study, a new PCR method for the detection of bacteriophages of O. oeni without the need for a sensitive indicator strain and for a growth step on solid medium to confirm the presence of prophages was developed.Until now, typing of O. oeni bacteriophages has been conducted by morphological characterization, structural-protein composition, host range analysis, and/or restriction enzyme analysis (5, 12-15). The last approach (15) was the first DNA-based technique that allowed the grouping of different O. oeni phage types.Bacteriophages of O. oeni present in wines were first observed by Sozzi et al. (12) by electron microscopy (EM) and were subsequently described as being different in size (5). Later, EM analyses of four O. oeni phages showed that they were very similar, exhibiting a Bradley type B morphology, with the presence of a head, a tail, and a base plate (13). Similar results were reported after EM analysis of 10 phages isolated from wines (14), which also showed similar morphology with slight differences in the head size, tail length, and presence of a base plate. These phages were divided into four groups based on structural-protein composition, host range, and restriction enzyme analysis. Likewise, comparable morphologies and structural protein profiles have also been reported in 17 O. oeni mitomycin C (mit C)-induced phages from different O. oeni strains, which were further divided into six groups based on their enzymatic restriction patterns (15).Endolysins, also termed lysins, are enzymes encoded by most double-stranded DNA phage genomes that lyse bacterial cell walls and cause host death (16,17). Detection of the lys gene has recently been used to evaluate phages infecting Lactobacillus helve-

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Typing of bacteriophages by randomly amplified polymorphic DNA (RAPD)-PCR to assess genetic diversity

Cited by 66 publications

References 33 publications

Isolation and Characterization of Listeria monocytogenes Bacteriophages from Environmental Sources in Kars-Turkey

Isolation and Characterization of Listeria monocytogenes Bacteriophages from Environmental Sources in Kars-Turkey

Biodiversity of Bacteriocin-Producing Lactic Acid Bacteria from Mexican Regional Cheeses and their Contribution to Milk Fermentation

Development of a New Method for Detection and Identification of Oenococcus oeni Bacteriophages Based on Endolysin Gene Sequence and Randomly Amplified Polymorphic DNA

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