scholarly journals Rep PCR Characterization of Lactic Acid Bacteria Isolated From Raw Cows’ Milk in Algeria.

Author(s):  
Akil LOMANI ◽  
Dennis S Nielsen ◽  
Larbi Ahmed Amine ◽  
Tahri Ahmed ◽  
Mediani Ahmed

Abstract Milk and indigenous fermented, milk based products are a rich source of lactic acid bacteria (LAB) and may serve as a source of potential probiotics. In the present study LAB were isolated from raw cows’ milk sampled at five different Algerian farms. A total of 24 Gram positive, catalase negative isolates where identified to species level using a combination of (GTG) 5-based rep-PCR fingerprinting and 16S rRNA gene sequencing. All isolates were identified as Enterococcus faecium and all isolates had highly similar rep-PCR profiles. Four representative isolates were screened for acid tolerance, bile salts tolerance, antimicrobial susceptibility, antibacterial activity and haemolysis. The four selected strains all exhibited good tolerance to low pH (2, 3, and 4), and to bile salts (concentrations of 0.5%, 1%, and 2%) and were sensitive to chloramphenicol, vancomycin, tetracycline, gentamicine and peniciline G, but were resistant to oxaciline. Cell-free supernatants of the four tested strains all inhibited Staphylococcus aureus, Escherichia coli and Listeria monocytogenes but not Salmonella Typhi. No haemolytic activity was observed.

2021 ◽  
Vol 11 (17) ◽  
pp. 7897
Author(s):  
Giuseppe Aprea ◽  
Alessandra Alessiani ◽  
Franca Rossi ◽  
Lorena Sacchini ◽  
Arianna Boni ◽  
...  

This study focused on the characterization of lactic acid bacteria (LAB) in a renowned traditional Italian cheese, Pecorino di Farindola, in order to select an autochthonous culture and investigate its potential for the improvement of safety and functional properties. Two hundred and six LAB isolated throughout production and maturation from nine cheese lots of three farms were identified by 16S rRNA gene sequencing and tested for the presence of genes encoding virulence factors, vancomycin resistance (for enterococci), biogenic amines (BAs) and bacteriocin production for antimicrobial activity; and for the capacity to survive in the gastrointestinal tract (GIT) based on tolerance to low pH and bile salts and adhesion to CaCo-2 cells. A Lacticaseibacillus paracasei isolate was used in cheese making and determined a decline of spiked Listeria monocytogenes and Escherichia coli O157 faster than in the control cheese. The autochthonous bacterial groups were numerically unaffected, apart from lactobacilli that were recovered in higher numbers in cheese with the addition of the L. paracasei strain. Based on repetitive extragenic palyndrome (Rep) polymerase chain reaction (PCR) profiles, the added strain possibly dominated until day 88 in the cheese. Results encourage further trials with autochthonous cultures in order to efficiently inhibit hazardous bacteria and to enrich a functional microbiota in Pecorino di Farindola and similar cheeses.


Author(s):  
Giuseppe Aprea ◽  
Alessandra Alessiani ◽  
Franca Rossi ◽  
Lorena Sacchini ◽  
Arianna Boni ◽  
...  

This study focused on the characterization of lactic acid bacteria (LAB) in a renowned traditional Italian cheese, Pecorino di Farindola, to select an autochthonous culture and investigate its po-tential for the improvement of safety and functional properties. Two hundred and six LAB isolated throughout production and maturation from nine cheese lots of three farms were identified by 16S rRNA gene sequencing and tested for the presence of genes encoding virulence factors, vancomycin resistance (for enterococci), biogenic amines (BAs) and bacteriocin production, for antimicrobial activity, and for the capacity to survive in the gastro-intestinal tract (GIT) based on tolerance to low pH and bile salts and adhesion to CaCo-2 cells. A Lacticaseibacillus paracasei isolate was used in cheese making and determined a decline of spiked Listeria monocytogenes and Escherichia coli O157 faster than in control cheese. The autochthonous bacterial groups were numerically unaffected, apart from lactobacilli that were recovered in higher numbers in the cheese with added L. paracasei. Based on Repetitive Extragenic Palyndrome (Rep) PCR profiles, the added strain possibly dominated until day 88 in cheese. Results encourage further trials with autochthonous cultures to efficiently inhibit hazardous bacteria and enrich a functional microbiota in Pecorino di Farindola and similar cheeses.


2019 ◽  
Vol 4 (1) ◽  
pp. 106-109
Author(s):  
Helen J. Lawalata ◽  
Jovialine A. Rungkat

Diversity of Lactic acid bacteria (LAB) that have ability to improved number of carnocine on bakasang were determind by application of molecular approach. Molecular characterization of LAB was based on 16S rRNA gene sequencing.  The result  showed that one isolate lactic acid bacteria (Pediococcus sp.B.5.1)   among 9 isolates LAB exhibited the highest ability to improved number of carnocine. The molecular characterization based on 16S rRNA gene sequence showed that the Pediococcus B.5.1 isolate  clearly belonged to members of species Pediococcus acidilactici. Therefore, lactic acid bacteria with the high ability to improved number of carnosine could be found from fermented fish bakasang and make bakasang as a functional food.


Author(s):  
Jinghui Yao ◽  
Jing Gao ◽  
Jianming Guo ◽  
Hengan Wang ◽  
En Zhang ◽  
...  

The consumption of cheese in China is increasing rapidly. Little is known about the microbiota, the presence of antibiotic-resistant bacteria, or the distribution of antibiotic resistance genes (ARGs) in commercially-produced cheeses sold in China. These are important criteria for evaluating quality and safety. Thus, this study assessed the metagenomics of fifteen types of cheese using 16S rRNA gene sequencing. Fourteen bacterial genera were detected. Lactococcus , Lactobacillus , and Streptococcus were dominant based on numbers of sequence reads. Multidrug-resistant lactic acid bacteria were isolated from most of the types of cheese. The isolates showed 100% and 91.7% resistance to streptomycin and sulfamethoxazole, respectively, and genes involved in acquired resistance to streptomycin ( strB) and sulfonamides ( sul2) were detected with high frequency. To analyze the distribution of ARGs in the cheeses in overall, 309 ARGs from eight categories of ARG and nine transposase genes were profiled. A total of 169 ARGs were detected in the 15 cheeses; their occurrence and abundance varied significantly between cheeses. Our study demonstrates that there is various diversity of the bacteria and ARGs in cheeses sold in China. The risks associated with multidrug resistance of dominant lactic acid bacteria are of great concern.


2020 ◽  
Vol 25 (6) ◽  
pp. 2079-2085
Author(s):  
LIN-HU NAN ◽  
◽  
YI-SHENG CHEN ◽  
HUI-CHUNG WU ◽  
YU-CHING SU ◽  
...  

Lychee is a popular fruit in China and southeastern Asia. Although it is very popular, the microbiota of lactic acid bacteria (LAB) associated with lychee remains poorly described. Lychee samples from seven different markets located in three cities in Taiwan were collected and a total of 104 LAB were isolated. Through RFLP analyses of 16S rDNA and rpoA genes for grouping and 16S rRNA gene sequencing, these isolates were finally divided into 6 groups (A to F). The most common genera of LAB in lychee samples were Weissella and Leuconostoc. Weissella confusa strain E was found to produce a bacteriocin active against Listeria monocytogenes and some other Gram-positive bacteria. Mass spectrometry analysis revealed the bacteriocin mass to be approximately 3426.77 Da, which is different to other known Weissella bacteriocins. In addition, strain MB7 included in the genus Leuconostoc was identified as potential novel species or subspecies on the basis of phylogenetic analysis of 16S rRNA, rpoA and pheS gene sequences. Thus, this is the first report describing the distribution and varieties of LAB associated with lychee fruits. In addition, one potential novel LAB species or subspecies and one potential novel bacteriocin were also reported in this study.


2019 ◽  
Vol 8 (1) ◽  
pp. 5 ◽  
Author(s):  
Anshul Sharma ◽  
Jasmine Kaur ◽  
Sulhee Lee ◽  
Young-Seo Park

The present work aimed at tracking intentionally inoculated lactic acid bacteria (LAB) strains in yogurt and probiotic powder. Leuconostoc (Leu.) mesenteroides (11251), Lactobacillus (L.) brevis (B151), and Lactobacillus plantarum (LB41K) strains were tracked in yogurt, and L. plantarum (LB41P) was tracked in a commercial probiotic powder. The yogurt was intentionally inoculated with the selected bacterial strains. Two types of yogurt with known and unknown bacterial pools were utilized. The standard 16S rRNA gene sequencing was used to evaluate the initial screening. The molecular typing tools, random amplified polymorphic DNA (RAPD), repetitive element palindromic PCR (rep-PCR), and comparative gene sequence analysis of selected housekeeping loci were used to track the inoculated dubious strains. Out of 30 random selections for each inoculation, the developed method identified seven (11251), nine (B151), and five (LB41K) colonies in the yogurt. The validation was performed by identifying 7 colonies (LB41P) out of 30 in the probiotic powder. The DNA banding profiles and the gene sequence alignments led to the identification of the correct inoculated strains. Overall, the study summarizes the use of molecular tools to identify the deliberately inoculated LAB strains. In conclusion, the proposed polyphasic approach effectively tracked the intentionally inoculated strains: Leu. mesenteroides, L. brevis, and L. plantarum (LB41K) in yogurt and L. plantarum (LB41P) in probiotic powder. The study demonstrates how to track industrially relevant misused LAB strains in marketable food products.


2017 ◽  
Vol 1 (1) ◽  
pp. 89-90 ◽  
Author(s):  
Hanna Aljaksandrauna Bareika ◽  
Anatasija Vyachaslavauna Sidarenka ◽  
Galina Ivanovna Novik

Abstract From fresh and fermented vegetables (cucumber, sauercraut, eggplant), cereals (wheat, rice), legumes (soy) 37 cultures of lactic acid bacteria were isolated. Based on biochemical characteristics, MALDI-TOF MS protein profiling and 16S rRNA gene sequencing they were identified as representatives of Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Enterococcus genera. Six members of Lactobacillus genera and two members of Leuconostoc genera actively producing EPS were selected for further investigation as components of starters for production of special dietetic foodstuffs.


2013 ◽  
Vol 14 (3) ◽  
Author(s):  
Rofiq Sunaryanto ◽  
Bambang Marwoto

Dadih is one of the traditional fermented milk products of West Sumatera. Dadih contain a lot of lactic acid bacteria that acts as a coagulant and preservative. Some lactic acid bacteria also act as a probiotic agent because of characteristics that are resistant to acidic conditions. Some of the main requirements of microbes that can be used as probiotic microbes were resistant to low pH, bile salts, able to colonize, and having antimicrobial activity. Each species of the genus Lactobacillus havedifferent characteristics. This characteristic are influenced by the environment in which the bacteria live. Isolation, identification and characterization of lactic acid bacteria derived from buffalo milk were done. The results of isolated lactic acid bacteria was Lactobacillus plantarum. The characterization of Lactobacillus plantarum indicated that these isolates were able to survive in a concentration of medium containing 0.5% bile salts, resistant to acidic media until pH 2, have antimicrobial activity (inhibit Escherichia coli, Staphylococcus aureus and Enterococcus faecalis).


Foods ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 489 ◽  
Author(s):  
Adriana Antunes-Rohling ◽  
Silvia Calero ◽  
Nabil Halaihel ◽  
Pedro Marquina ◽  
Javier Raso ◽  
...  

The aim of this study was to characterize the spoilage microbiota of hake fillets stored under modified atmospheres (MAP) (50% CO2/50% N2) at different temperatures using high-throughput 16S rRNA gene sequencing and to compare the results with those obtained using traditional microbiology techniques. The results obtained indicate that, as expected, higher storage temperatures lead to shorter shelf-lives (the time of sensory rejection by panelists). Thus, the shelf-life decreased from six days to two days for Batch A when the storage temperature increased from 1 to 7 °C, and from five to two days—when the same increase in storage temperature was compared—for Batch B. In all cases, the trimethylamine (TMA) levels measured at the time of sensory rejection of hake fillets exceeded the recommended threshold of 5 mg/100 g. Photobacterium and Psychrobacter were the most abundant genera at the time of spoilage in all but one of the samples analyzed: Thus, Photobacterium represented between 19% and 46%, and Psychrobacter between 27% and 38% of the total microbiota. They were followed by Moritella, Carnobacterium, Shewanella, and Vibrio, whose relative order varied depending on the sample/batch analyzed. These results highlight the relevance of Photobacterium as a spoiler of hake stored in atmospheres rich in CO2. Further research will be required to elucidate if other microorganisms, such as Psychrobacter, Moritella, or Carnobacterium, also contribute to spoilage of hake when stored under MAP.


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