Background: Escherichia coli infections are one of the major health problems in many countries. The emergence of antibiotic resistance has complicated the treatment of infections caused by this microorganism. Today, researchers are focusing on novel approaches for the treatment of E. coli infections among which is using spore-forming probiotics for their high stability in the harsh gastrointestinal (GIT) environment. Results: This study screened the potential of B. coagulans and B. subtilis to inhibit the expression of flu, eae, luxS, and ctxM genes in 40 E. coli isolated from a total of 300 fecal samples of patients suffering from diarrhea from August 2018 to January 2019. For this purpose, first the presence of these genes was confirmed in the isolates using the PCR method. Then, the standard strains and broiler-derived B. subtilis and B. coagulans were used to screen their ability to tolerate low pH, bile salts, and gastric enzymes. After confirming their ability to survive under the simulant gastric environment, they were cocultured with E. coli isolates (n=4) harboring all the flu, eae, luxS, and ctxM genes. After reaching the logarithmic growth phase, the expression levels of flu, eae, luxS, and ctxM genes were determined using the real-time PCR methods. According to the results, there was a statistically significant relationship between the exposure of E. coli with commercial and broiler-derived spore-forming probiotics and the reduced expression of these genes. Conclusion: The broiler-derived isolates had a greater capacity to decrease the expression of these genes compared to the standard strains, proposing their adoption for dietary supplementations. However, more studies are required to study the effects of these spore-forming bacteria on E. coli infections in vivo and their possible influence on lactose tolerance, nutritional absorption, and reduced cholesterol levels.
Background Escherichia coli antibiotic resistance is one of the major health problems in many countries. Nowadays, researchers are focusing on novel approaches for the treatment of E. coli infections, including the use of spore-forming probiotics for their high stability in the harsh gastrointestinal (GIT) environment.MethodsInitially, 300 stool samples were collected from patients with gastrointestinal infections admitted to Imam Khomeini Hospital in Tehran. Then, diagnostic tests were performed to detect E. coli isolates on the samples. A DNA test was applied to examine the presence of ctxM, luxS, eae and flu in the samples. Afterward, the effect of native and commercial probiotics of Bacillus subtilis and Bacillus coagulant was investigated on the expression of the studied genes. ResultsGenes of flu, eae, luxS, and ctxM which are involved in bacterial attachment, biofilm formation, signaling, and antimicrobial resistance were existing in four out of 40 E. coli isolated among patients suffering from diarrhea. The expression levels of flu, luxS, eae, and ctxM genes decreased significantly (p-value < 0.05) after co-culture of E. coli with B. subtilis ATCC 6633 and broiler-derived B. subtilis. Both broiler-derived B. subtilis and broiler-derived B. coagulans have a significant effect on all the studied genes.Conclusions The broiler-derived isolates had a greater capacity to decrease the expression of these genes than the standard strains, proposing their adoption for dietary supplementations.
Background and objective: Antibiotics are widely used worldwide. However, due to the emergence of antibiotic resistance in a wide range of microorganisms, their use worldwide has failed. Probiotics are suggested as complementary and alternative medicine. The present study aimed to investigate the effects of probiotics isolated from local chickens on the expression of luxS and ctxM genes in resistant Escherichia coli. Materials and methods: 300 fecal samples were taken from patients referring to Imam Khomeini Hospital in Tehran during May–September 2016 and Escherichia coli samples were isolated using specific culture media and biochemical tests and then the presence of luxS and ctxM genes were identified using PCR with specific primers. In order to extract the probiotics forming spores, the intestinal contents of 10 poultry that had not used any antibiotics and probiotics were cultured, isolated, and identified using biochemical and PCR methods. Commercial strains of Bacillus subtilis and Bacillus coagulans were purchased to compare their effects with native bacteria. These strains were then co-cultured with resistant Escherichia coli strains containing ctxM and luxS genes. Real-time PCR was used to evaluate the effect of these probiotics on gene expression Results: The results indicated that 40 isolates (7.5%) of Escherichia coli were obtained from the 300 fecal samples. Thirteen samples (32.5%) were outpatients and 27 (67.5%) were inpatients. All isolates were isolated from men and women aged 21–62. Four Escherichia coli strains were isolated from patients carrying ctxM and luxS genes. Isolation of Bacillus coagulans and Bacillus subtilis from samples was confirmed by biochemical and molecular experiments. The commercial and native strains of Bacillus coagulans reduced the expression of the luxS and ctxM genes by 3.60, 3.30, 1.58, and 2.70 times respectively. Also, the commercial and native strains of Bacillus subtilis decreased the expression of the luxS and ctxM genes by 1.37, 1.10, 2.20, and 2.80 times respectively. The results of statistical analysis showed a significant relationship between the presence of native and commercial probiotics in culture and reduced expression of ctxM and luxS genes. Conclusion: According to the results, supplements of Bacillus coagulans and Bacillus subtilis increase the effect of antibiotics resistance in Escherichia coli by reducing the expression of resistance genes.
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