Detection of culturable and viable but non-culturable cells of beer spoilage lactic acid bacteria by combined use of propidium monoazide andhorA-specific polymerase chain reaction

2016 ◽  
Vol 122 (1) ◽  
pp. 29-33 ◽  
Author(s):  
Yang Deng ◽  
Junfeng Zhao ◽  
Huiping Li ◽  
Zhenbo Xu ◽  
Junyan Liu ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Propidium Monoazide ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Combined Use ◽  
Beer Spoilage ◽  
Polymerase Chain

scholarly journals Inter- and Intra-Species Diversity of Lactic Acid Bacteria in Apis mellifera ligustica Colonies

2020 ◽  
Vol 8 (10) ◽  
pp. 1578 ◽  
Author(s):  
Massimo Iorizzo ◽  
Gianfranco Pannella ◽  
Silvia Jane Lombardi ◽  
Sonia Ganassi ◽  
Bruno Testa ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lactic Acid ◽  
Apis Mellifera ◽  
Lactic Acid Bacteria ◽  
Honey Bees ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Rapd Pcr ◽  
The Social ◽  
Polymerase Chain

Lactic acid bacteria could positively affect the health of honey bees, including nutritional supplementation, immune system development and pathogen colonization resistance. Based on these considerations the present study evaluated predominant Lactic Acid Bacteria (LAB) species from beebread as well as from the social stomach and midgut of Apis mellifera ligustica honey bee foragers. In detail, for each compartment, the diversity in species and biotypes was ascertained through multiple culture-dependent approaches, consisting of Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE), 16S rRNA gene sequencing and Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR). The study of a lactic acid bacteria community, performed with PCR-DGGE and sequence analysis targeting the V1–V3 region of the 16S rRNA gene (rDNA), highlighted the presence of a few species, including Apilactobacillus kunkeei, Lactiplantibacillus plantarum, Fructobacillus fructosus, Levilactobacillus brevis and Lactobacillus delbrueckii subsp. lactis. Depending on the different compartments, diverse levels of biodiversity in species were found. Particularly, a very low inter-species biodiversity was detected in the midgut that was prevalently dominated by the presence of Apilactobacillus kunkeei. On the other hand, the beebread was characterized by a reasonable biodiversity showing the presence of five species and the predominance of Apilactobacillus kunkeei, Lactiplantibacillus plantarum and Fructobacillus fructosus. The RAPD-PCR analysis performed on the three predominant species allowed the differentiation into several biotypes for each species. Moreover, a relationship between biotypes and compartments has been detected and each biotype was able to express a specific biochemical profile. The biotypes that populated the social stomach and midgut were able to metabolize sugars considered toxic for bees while those isolated from beebread could contribute to release useful compounds with functional properties. Based on this knowledge, new biotechnological approaches could be developed to improve the health of honey bees and the quality of bee products.

scholarly journals Isolation and Identification of Lactic Acid Bacteria from Natural Whey Cultures of Buffalo and Cow Milk

Foods ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 233
Author(s):  
Rosangela Marasco ◽  
Mariagiovanna Gazzillo ◽  
Nicoletta Campolattano ◽  
Margherita Sacco ◽  
Lidia Muscariello
Keyword(s):  
Polymerase Chain Reaction ◽  
Lactic Acid ◽  
Microbial Community ◽  
Lactic Acid Bacteria ◽  
Microbial Diversity ◽  
Strain Level ◽  
Sequencing Analysis ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Polymerase Chain

In southern Italy, some artisanal farms produce mozzarella and caciocavallo cheeses by using natural whey starter (NWS), whose microbial diversity is responsible for the characteristic flavor and texture of the final product. We studied the microbial community of NWS cultures of cow’s milk (NWSc) for the production of caciocavallo and buffalo’s milk (NWSb) for the production of mozzarella, both from artisanal farms. Bacterial identification at species and strain level was based on an integrative strategy, combining culture-dependent (sequencing of the 16S rDNA, species/subspecies-specific Polymerase Chain Reaction (PCR) and clustering by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) and culture-independent (next-generation sequencing analysis, NGS) approaches. Results obtained with both approaches showed the occurrence of five species of lactic acid bacteria in NWSb (Lactococcus lactis subsp. lactis, Lactobacillus fermentum, Streptococcus thermophilus, Lactobacillus delbrueckii, and Lactobacillus helveticus) and five species in NWSc (Lc. lactis subsp. lactis, Enterococcus faecium, and S. thermophilus, Lb. helveticus, and Lb. delbrueckii), with the last two found only by the NGS analysis. Moreover, RAPD profiles, performed on Lc. lactis subsp. lactis different isolates from both NWSs, showed nine strains in NWSb and seven strains in NWSc, showing a microbial diversity also at strain level. Characterization of the microbiota of natural whey starters aims to collect new starter bacteria to use for tracing microbial community during the production of artisanal cheeses, in order to preserve their quality and authenticity, and to select new Lactic Acid Bacteria (LAB) strains for the production of functional foods.

A specific oligonucleotide primer for the rapid detection of Lactobacillus lindneri by polymerase chain reaction

1997 ◽  
Vol 43 (2) ◽  
pp. 157-163 ◽  
Author(s):  
Tetsuji Yasui ◽  
Hiroshi Taguchi ◽  
Tomoyuki Okamoto
Keyword(s):  
Polymerase Chain Reaction ◽  
Lactic Acid ◽  
16S Rdna ◽  
Rapid Detection ◽  
Oligonucleotide Primer ◽  
Published Data ◽  
Specific Primer ◽  
Chain Reaction ◽  
Beer Spoilage ◽  
Polymerase Chain

A polymerase chain reaction (PCR) method was developed for the rapid detection of the beer-spoilage heterofermentative lactic acid bacterium Lactobacillus lindneri. Three strains, the Chinese brewery isolate DA1, the Japanese commercial beer isolate BG2, and the Japanese brewery isolate SE3, which were serologically classified as belonging to L. lindneri, were used in this study. After sequencing the 16S rDNA of the isolates DA1 and BG2 and the typical beer-spoilage heterofermentative Lactobacillus brevis L63, these sequences were compared with published data. A L. lindneri specific PCR primer, DA-40, was then constructed based on the V1 variable region of 16S rDNA. The specificity of PCR using the L. lindneri specific primer DA-40 and the universal primer 907r was examined using five L. lindneri strains: the three isolates described above and two strains from culture collection, DSM 20690 and DSM 20692. A variety of beer-spoilage lactic acid bacteria, including 71 Lactobacillus strains and 13 Pediococcus strains, were also included in this examination. No PCR product was obtained from any DNA with the exception of the five L. lindneri strains, indicating that the L. lindneri specific primer DA-40 was highly specific. The detection limit for L. lindneri in beer was 63 CFU/100 mL of beer.Key words: Lactobacillus lindneri, oligonucleotide probe, sequence analysis, 16S rDNA.

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