scholarly journals Novel Approach to Mycotoxin Detoxification in Farm Animals Using Probiotics Added to Feed Stuffs

2010 ◽  
Author(s):  
Roni Shapira ◽  
Judith Grizzle ◽  
Nachman Paster ◽  
Mark Pines ◽  
Chamindrani Mendis-Handagama
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
High Efficiency ◽  
Farm Animals ◽  
Probiotic Bacteria ◽  
Expression Vectors ◽  
Growth Media ◽  
Strong Promoter ◽  
E Coli ◽  
Major Interest

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.

Biological Detoxification of the Mycotoxin Deoxynivalenol (DON) to Improve Safety of Animal Feed and Food

2010 ◽  
Author(s):  
Ting Zhou ◽  
Roni Shapira ◽  
Peter Pauls ◽  
Nachman Paster ◽  
Mark Pines
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
High Efficiency ◽  
Fluorescent Protein ◽  
Animal Feed ◽  
Probiotic Bacteria ◽  
Expression Vectors ◽  
Growth Media ◽  
Strong Promoter ◽  
E Coli

The trichothecene deoxynivalenol (DON, vomitoxin), one of the most common mycotoxin contaminants of grains, is produced by members of the Fusarium genus. DON poses a health risk to consumers and impairs livestock performance because it causes feed refusal, nausea, vomiting, diarrhea, hemolytic effects and cellular injury. 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 overall objective 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.

Effect of bovine colostrum, cheese whey, and spray-dried porcine plasma on the in vitro growth of probiotic bacteria and Escherichia coli

2014 ◽  
Vol 60 (5) ◽  
pp. 287-295 ◽  
Author(s):  
Claude P. Champagne ◽  
Yves Raymond ◽  
Yves Pouliot ◽  
Sylvie F. Gauthier ◽  
Martin Lessard
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
Cheese Whey ◽  
Probiotic Bacteria ◽  
E Coli ◽  
Spray Dried ◽  
Mrs Broth

The aim of this study is to evaluate the effects of defatted colostrum (Col), defatted decaseinated colostrum whey, cheese whey, and spray-dried porcine plasma (SDPP) as supplements of a growth medium (de Man – Rogosa – Sharpe (MRS) broth) on the multiplication of lactic acid bacteria, probiotic bacteria, and potentially pathogenic Escherichia coli. Using automated spectrophotometry (in vitro system), we evaluated the effect of the 4 supplements on maximum growth rate (μmax), lag time (LagT), and biomass (ODmax) of 12 lactic acid bacteria and probiotic bacteria and of an E. coli culture. Enrichment of MRS broth with a Col concentration of 10 g/L increased the μmax of 5 of the 12 strains by up to 55%. Negative effects of Col or SDPP on growth rates were also observed with 3 probiotic strains; in one instance μmax was reduced by 40%. The most effective inhibitor of E. coli growth was SDPP, and this effect was not linked to its lysozyme content. The positive effect of enrichment with the dairy-based ingredient might be linked to enrichment in sugars and increased buffering power of the medium. These in vitro data suggest that both Col and SDPP could be considered as supplements to animal feeds to improve intestinal health because of their potential to promote growth of probiotic bacteria and to inhibit growth of pathogenic bacteria such as E. coli.

scholarly journals Antimicrobial Potential of Probiotics in Combination with Start er Lactic Acid Bacteria

2017 ◽  
Vol 48 (4) ◽  
pp. 208-215 ◽  
Author(s):  
T. Mančušková ◽  
A. Medveďová ◽  
Ľ. Valík
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Acidophilus ◽  
Probiotic Bacteria ◽  
E Coli ◽  
Antimicrobial Potential ◽  
Final Density ◽  
Average Change ◽  
Lactobacillus Acidophilus Ncfm

Abstract The symbiotic interrelationship of probiotic bacteria and starter lactic acid bacteria is of fundamental importance in many fields of industrial microbiology and it is also a great interest of food technologists. This study deals with the antimicrobial potential of cell-free supernatants of Lactobacillus acidophilus NCFM, L. rhamnosus GG and Fresco culture when cultivated alone or together against 5 strains of Escherichia coli and 5 strains of Staphylococcus aureus. While the effect on E. coli was not proven (average change of final density compared to the control was only ΔNEC,24h = 0.12 log CFU ml-1), the decrease of S. aureus final density in the presence of nisin and cell-free supernatant of L. acidophilus NCFM and Fresco culture was considerable (ΔNSA,24h,nis10 = -1.95 log CFU ml-1 and ΔNSA,24h,NCFM+Fr24 = -0.69 log CFU ml-1, respectively).

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
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Farm Animals ◽  
Fish Analysis ◽  
Culture Methods ◽  
Associated Bacteria ◽  
E Coli ◽  
Bacterial Groups

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.

scholarly journals Complete Genome Sequence of Lactobacillus reuteri Byun-re-01, Isolated from Mouse Small Intestine

2018 ◽  
Vol 7 (17) ◽  
Author(s):  
Dongjun Kim ◽  
Mun-ju Cho ◽  
Seungchan Cho ◽  
Yongjun Lee ◽  
Sung June Byun ◽  
...  
Keyword(s):  
Small Intestine ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Lactobacillus Reuteri ◽  
Probiotic Bacteria ◽  
Content Type ◽  
Mouse Small Intestine

Lactic acid bacteria (LAB) are generally recognized as safe (GRAS) and serve as probiotic bacteria when consumed in adequate amounts. Here, we report the complete genome sequence of Lactobacillus reuteri Byun-re-01, isolated from mouse small intestine.

scholarly journals Lactic Acid Bacteria (Lactobacillus bulgaricus and Streptococcus thermophillus) in Yogurt Inhibit the Growth of Escherichia coli, Salmonella typhimurium, and Shigella sp. In Vitro

2021 ◽  
Vol 31 (4) ◽  
pp. 2
Author(s):  
IDSAP Peramiarti
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Salmonella Typhimurium ◽  
Pathogenic Bacteria ◽  
Test Method ◽  
P Value ◽  
E Coli ◽  
Significant Difference

Diarrhea is defecation with a frequency more often than usual (three times or more) a day (10 mL/kg/day) with a soft or liquid consistency, even in the form of water alone. Pathogenic bacteria, such as Escherichia coli, Salmonella typhimurium, and Shigella sp., play a role in many cases, to which antibiotics are prescribed as the first-line therapy. However, since antibiotic resistance cases are often found, preventive therapies are needed, such as consuming yogurt, which is produced through a fermentation process by lactic acid bacteria (LAB). This research aimed to determine the activity of lactic acid bacteria (Liactobacillus bulgaricus and Streptococcus thermophilus) in yogurt in inhibiting the growth of the pathogenic bacteria E. coli, S. typhimurium, and Shigella sp. The research applied in vitro with the liquid dilution test method and the true experimental design research method with post-test-only and control group design. The design was used to see the inhibitory effect of yogurt LAB on the growth of E. coli, S. typhimurium, and Shigell sp. to compare the effect of several different yogurt concentrations, namely 20%, 40%, 60%, and 80%. The results of the Least Significance Different analysis showed that there was a significant difference between yogurt with a concentration of 0% and that with various concentrations in inhibiting the growth of E. coli, S. typhimurium, and Shigella sp. with a p-value of &lt;0.05. Whereas, there was no significant difference in the various concentrations of yogurt in inhibiting the growth of the three kinds of bacteria with a p-value of &gt; 0.05.<p class="Default" align="center"> </p>

scholarly journals Viability of lactic acid bacteria in different components of Ogi with anti diarrhoeagenic E. coli activities

2019 ◽  
Vol 3 (6) ◽  
pp. 206-213
Author(s):  
Roseline Eleojo Kwasi ◽  
Iyanuoluwa Gladys Aremu ◽  
Qudus Olamide Dosunmu ◽  
Funmilola A. Ayeni
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Antimicrobial Properties ◽  
Culture Method ◽  
Antimicrobial Activities ◽  
Drying Methods ◽  
Bacterial Strains ◽  
E Coli ◽  
Log Reduction ◽  
Freeze Dried

Background: Ogi constitutes a rich source of lactic acid bacteria (LAB) with associated health benefits to humans through antimicrobial activities. However, the high viability of LAB in Ogi and its supernatant (Omidun) is essential. Aims: This study was carried out to assess the viability of LAB in various forms of modified and natural Ogi and the antimicrobial properties of Omidun against diarrhoeagenic E coli. Methods and Material: The viability of LAB was assessed in fermented Ogi slurry and Omidun for one month and also freeze-dried Ogi with and without added bacterial strains for two months. A further 10 days viability study of modified Omidun, refrigerated Omidun, and normal Ogi was performed. The antimicrobial effects of modified Omidun against five selected strains of diarrhoeagenic E. coli (DEC) were evaluated by the co-culture method. Results: Both drying methods significantly affected carotenoids and phenolic compounds. The Ogi slurry had viable LAB only for 10 days after which, there was a succession of fungi and yeast. Omidun showed 2 log10cfu/ml reduction of LAB count each week and the freeze-dried Ogi showed progressive reduction in viability. Refrigerated Omidun has little viable LAB, while higher viability was seen in modified Omidun (≥2 log cfu/ml) than normal Omidun. Modified Omidun intervention led to 2-4 log reduction in diarrhoeagenic E. coli strains and total inactivation of shigella-toxin producing E. coli H66D strain in co-culture. Conclusions: The consumption of Ogi should be within 10 days of milling using modified Omidun. There are practical potentials of consumption of Omidun in destroying E. coli strains implicated in diarrhea. Keywords: Ogi, Omidun, lactic acid bacteria, diarrhoeagenic Escherichia coli strains, Viability.

scholarly journals Antimicrobial activity of lactic acid bacteria isolated from garri on Escherichia coli strains isolated from clinical and environmental samples

2020 ◽  
Vol 12 (4) ◽  
pp. 357-365
Author(s):  
H.I. Atta ◽  
A. Gimba ◽  
T. Bamgbose
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactobacillus Acidophilus ◽  
Plant Pathogens ◽  
Resistant Bacteria ◽  
Fermented Foods ◽  
Environmental Isolates ◽  
E Coli

Abstract. The production of bacteriocins by lactic acid bacteria affords them the ability to inhibit the growth of bacteria; they are particularly important in the biocontrol of human and plant pathogens. Lactic acid bacteria have been frequently isolated from fermented foods due to the high acidity these foods contain. In this study, lactic acid bacteria were isolated from garri, a popular Nigerian staple food, which is fermented from cassava, and their antagonistic activity against clinical and environmental isolates of Escherichia coli was determined. The species of Lactobacillus isolated include: Lactobacillus plantarum (50%), Lactobacillus fermentum (20%), Lactobacillus acidophilus (20%), and Lactobacillus salivarius (10%). Growth inhibition of the strains of E.coli was observed in Lactobacillus plantarum that inhibited the growth of both. The clinical and environmental isolates of E. coli were inhibited by Lactobacillus plantarum, while Lactobacillus acidophilus showed activity against only the clinical isolate. The greatest zone of inhibition against the strains of E. coli was recorded by Lactobacillus acidophilus (22.7±1.53 mm). The bacteriocins produced by Lactobacillus species have a good potential in the biocontrol of pathogens, and should be the focus of further studies on antibiotic resistant bacteria.

Pro-/Anti-Inflammatory Food Supplements

2020 ◽  
pp. 142-168
Author(s):  
Hüseyin Eseceli
Keyword(s):  
Immune Response ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Food Supplements ◽  
Gastrointestinal System ◽  
Probiotic Bacteria ◽  
Immune Systems ◽  
Positive Effects ◽  
Endogenous Defense ◽  
Probiotic Effect

Probiotics are living microorganisms, meaning “for life” and consist of two parts, “pro” and “biota,” and prebiotics are indigestible carbohydrates that increase the number and activities of colon bacteria and the effectiveness of probiotics. Probiotic consumption has been reported to have many positive effects, such as increasing immune response, balancing the colony, and increasing endogenous defense capacity of cells. The aim of prebiotics is to grow probiotic bacteria, thus improving the gastrointestinal and immune systems. Recent studies have shown that probiotics may have an impact on gastrointestinal system diseases. It has been shown to stimulate the proliferation of beneficial microorganisms in prebiotics and assist in the probiotic effect. The most commonly used probiotics and prebiotics are lactic acid bacteria that are types of Lactobacillus and Bifidobacterium. Prebiotics are naturally present in nutrients. Probiotics and prebiotics are used to prevent many diseases.

scholarly journals Microbiological Condition of Retail Beef Steaks: A Canadian Survey

2018 ◽  
Vol 7 (4) ◽  
pp. 1 ◽  
Author(s):  
Xianqin Yang ◽  
Julia Devos ◽  
Hui Wang ◽  
Mark Klassen
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Microbial Quality ◽  
Indicator Organisms ◽  
Mean Values ◽  
E Coli ◽  
Upper Limit ◽  
Brochothrix Thermosphacta ◽  
Beef Steaks

The second national baseline microbiological survey of beef steaks offered for retail in Canada was conducted in 2015. A total of 621 steaks of four types (cross rib, CR; inside round, IR; striploin, SL; top sirloin, TS) collected from 135 retail stores in five cities across Canada were tested. Swab samples each from swabbing the entire upper surface of each steak were processed for enumeration of seven groups of indicator organisms: total aerobes (AER), psychrotrophs (PSY), lactic acid bacteria (LAB), pseudomonads (PSE), Brochothrix thermosphacta (BRO), coliforms (COL) and Escherichia coli (ECO). The overall mean values (log CFU/100 cm2) were 5.17±1.29, 4.92±1.36, 4.79±1.42, 3.26±1.49, 2.34±1.88, and 0.80±1.05 for AER, PSY, LAB, PSE, BRO, and COL, respectively. ECO were not recovered from 87.3% of the steaks and when there was recovery, the numbers were mostly ≤ 1 log CFU/100 cm2. Strong correlation was found between the log numbers of AER and PSY, of AER and LAB, and of PSY and LAB, while the correlation between the log numbers of COL and ECO was weak. The numbers of COL and ECO from different groups of steak types or from different cities were not substantially different. Of the four types of steaks, IR had the lowest median values for AER, PSY, LAB, PSE and BRO, followed by CR. The microbiological condition of retail beef steaks in this survey was on par with that in the previous one, with very low numbers of generic E. coli being recovered from very few steaks and the indicators for microbial quality being at numbers much lower than the upper limit for shelf life of beef.

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