Characterization of the Gastrointestinal Mucosa–Associated Microbiota of Pigs and Chickens Using Culture-Based and Molecular Methodologies

2007 ◽  
Vol 70 (12) ◽  
pp. 2799-2804 ◽  
Author(s):  
M. CARMEN COLLADO ◽  
YOLANDA SANZ

The microbiota of the gastrointestinal tract (GIT) can play an important role in the healthy status of farm animals and in the safety of the whole food chain. In this study, the mucosa-associated microbiota of the GIT of pigs and chickens was analyzed by culture methods and fluorescence in situ hybridization combined with flow cytometry (FCM-FISH). In all pig GIT sections, lactic acid bacteria, Enterobacteriaceae, Atopobium, Bacteroides, and Clostridium histolyticum were the predominant bacterial groups. Atopobium, Bifidobacterium, Bacteroides, and Lactobacillus were detected at higher levels (P < 0.05) in the intestine than in the stomach. In all broilers' GIT sections, lactic acid bacteria, Atopobium, Bacteroides, and Escherichia coli were the predominant bacterial groups. Atopobium, Bifidobacterium, E. coli, and Eubacterium rectale–Clostridium coccoides counts were significantly higher (P < 0.05) in the crop, while those of Bacteroides and Lactobacillus were higher (P < 0.05) in the large intestine. Lactic acid bacteria were one of the predominant GIT mucosa–associated bacteria of pigs and broilers, which could be an index of their healthy status. FCM-FISH analysis also allowed the detection of bacterial groups hard to cultivate yet quantitatively important. The distribution of Lactobacillus and Bacteroides followed the same trend in both animal species, whereas that of Atopobium and Bifidobacterium was the opposite. These results contribute to the knowledge on the diversity and distribution of the animal GIT mucosa–associated microbiota.

Author(s):  
ROSALINA YULIANA AYEN ◽  
ENDANG KUSDIYANTINI ◽  
SRI PUJIYANTO

Objective: This research aimed to isolate, determine the characteristics of lactic acid bacteria (LAB) of Sui Wu’u from Bajawa, Nusa Tenggara Timur and identify LAB using 16S rRNA potential as antimicrobial activity against pathogenic bacteria. Methods: Sui Wu’u which has been stored for 6 months was obtained from Bajawa district, inoculated on de Man Rogosa-Sharpe Agar (Merck) + 0.5% CaCO3, purification of LAB, characterization of selected isolates, biochemical test, tolerance test for pH, viability to test temperature, and content NaCl, determination of antimicrobial action by the agar well disk diffusion method using antibiotic (Amoxicillin) as a control and as indicator bacteria (Staphylococcus aureus and Escherichia coli) and isolation of genomic 16S rRNA; molecular identification. Results: Based on research results obtained five isolates of LAB, Gram staining the LAB isolated from Sui Wu’u showed that the isolated bacteria (bacilli and coccus) are Gram-positive, catalase-negative and the isolates have tolerance of viability at temperatures of 10°C, 45°C, and 50°C and to salinitas of 4% and 6.5%. The inhibitory zone LAB isolates (2PKT) against E. coli bacteria (20 mm) and S. aureus (12 mm), and (2PKB) against E. coli bacteria (17 mm) and S. aureus (10 mm). The two selected isolates were identified as Lactobacillus fermentum strain HB bacteria with 100% identification value and 98.93% query cover and L. fermentum strain HT with 100% identification value and 99.23% query cover. Conclusion: L. fermentum from Sui Wu’u has antibacterial activity against Staphylococcus aureus and Escherichia coli.


2010 ◽  
Author(s):  
Roni Shapira ◽  
Judith Grizzle ◽  
Nachman Paster ◽  
Mark Pines ◽  
Chamindrani Mendis-Handagama

T-2 toxin, a toxic product belongs to the trichothecene mycotoxins, attracts major interest because of its severe detrimental effects on the health of human and farm animals. The occurrence of trichothecenes contamination is global and they are very resistant to physical or chemical detoxification techniques. Trichothecenes are absorbed in the small intestine into the blood stream. The hypothesis of this project was to develop a protecting system using probiotic bacteria that will express trichothecene 3-O-acetyltransferase (Tri101) that convert T-2 to a less toxic intermediate to reduce ingested levels in-situ. The major obstacle that we had faced during the project is the absence of stable and efficient expression vectors in probiotics. Most of the project period was invested to screen and isolate strong promoter to express high amounts of the detoxify enzyme on one hand and to stabilize the expression vector on the other hand. In order to estimate the detoxification capacity of the isolated promoters we had developed two very sensitive bioassays.The first system was based on Saccharomyces cerevisiae cells expressing the green fluorescent protein (GFP). Human liver cells proliferation was used as the second bioassay system.Using both systems we were able to prove actual detoxification on living cells by probiotic bacteria expressing Tri101. The first step was the isolation of already discovered strong promoters from lactic acid bacteria, cloning them downstream the Tri101 gene and transformed vectors to E. coli, a lactic acid bacteria strain Lactococcuslactis MG1363, and a probiotic strain of Lactobacillus casei. All plasmid constructs transformed to L. casei were unstable. The promoter designated lacA found to be the most efficient in reducing T-2 from the growth media of E. coli and L. lactis. A prompter library was generated from L. casei in order to isolate authentic probiotic promoters. Seven promoters were isolated, cloned downstream Tri101, transformed to bacteria and their detoxification capability was compared. One of those prompters, designated P201 showed a relatively high efficiency in detoxification. Sequence analysis of the promoter region of P201 and another promoter, P41, revealed the consensus region recognized by the sigma factor. We further attempted to isolate an inducible, strong promoter by comparing the protein profiles of L. casei grown in the presence of 0.3% bile salt (mimicking intestine conditions). Six spots that were consistently overexpressed in the presence of bile salts were isolated and identified. Their promoter reigns are now under investigation and characterization.


Author(s):  
L. Ketrouci ◽  
F. Dalache ◽  
D. Benabdelmoumene ◽  
A.A. Dahou ◽  
A. Homrani

Background: Technological characterization of lactic acid bacteria isolated from sheep’s milk collected in 3 regions in northwestern Algeria.Methods: During the period from 2018 to 2019, fifty strains of Lactic acid bacteria isolated from samples sheep’s milk were evaluated for several technologically-relevant properties: diacetyl and exopolysaccharides production, acidification, proteolytic and lipolytic activity and their antagonist activity against Escherichia coli and Pseudomonas aeruginosa. Result: The results indicate that among all the isolates only 20% were distinguished by their production of EPS mainly the genus Leuconostoc. Diacetyl production was observed in 71% of Lactobacillus, 60% of Enterococcus and 25% in Leuconostoc. 94% isolates showed moderate proteolytic activity. 56% and 60% of the strains degraded tween 80 and olive oil respectively for lipolytic activity. Inhibition activity by the cultures LAB was about 82% and 78% against E. coli and P. aeruginosa respectively. No culture supernatants inhibit P. aeruginosa, however 18% of the Enterococcus trains inhibit E. coli. BME1.A2 and BME2.D4 showed their highest acidification capacity developing a very large quantity of lactic acid after 24 h of incubation, i.e., 7.6 and 8.4 g lactic acid/L respectively.


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.


2021 ◽  
Vol 709 (1) ◽  
pp. 012020
Author(s):  
Evy Rossi ◽  
Akhyar Ali ◽  
Raswen Efendi ◽  
Fajar Restuhadi ◽  
Yelmira Zalfiatri ◽  
...  

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