Determining the role of a probiotic in the restoration of intestinal microbial balance by molecular and cultural techniques.
The human intestine has a vast variety of microorganisms, and their balance is dependent on several factors. Antibiotics affect microfloral balance and allow naturally opportunistic organisms to multiply. Azithromycin is the most widely used macrolide antibiotic, active against a wide number of pathogens including Pseudomonas aeruginosa and Staphylococcus aureus. It is currently used in the treatment of cystic fibrosis patients. The use of probiotics has advantages in gastrointestinal conditions, including infectious diarrhea and imbalance due to antibiotic use. In this research, the effect of azithromycin on the intestinal microbiota of Sprague Dawley rats and the role of Lactobacillus acidophilus in the restoration of the balance by employing molecular and cultural techniques was investigated. PCR with universal primers targeting the V3 region of the 16S rRNA gene followed by DGGE was used to characterize the overall intestinal microbiota composition. Cultivable fecal bacteria count using microbiological media and semi-quantitative PCR with group-specific primers were also utilized to analyze the effects of antibiotic and probiotic on microflora. We found that the total amount of 16S rRNA gene and fecal aerobic bacterial count was reduced following azithromycin administration along with elimination of non-pathogenic Escherichia coli, but it was restored by the use of the probiotic. The results from PCR with group-specific primers showed that Bacteroides sp was present in the control and probiotic groups, but it was nearly eliminated in the antibiotic group. Moreover, semi-quantitative PCR revealed that the numbers of Enterobacteriaceae were nearly the same in the probiotic group and decreased in the antibiotic group, while Bifidobacterium was significantly increased in the probiotic group and decreased in the antibiotic group (P < 0.05) as compared with that in the control group. Azithromycin-induced dysbiosis can result in prolonged deleterious effects on the host. The present study revealed that the use of lactic acid bacteria particularly L. acidophilus helped to restore intestinal microfloral balance.
Department of Biotechnology, Dalian Medical University, Dalian, China.
Affhan
A
Shoaib
Department of Biotechnology, Dalian Medical University, Dalian, China.
W
W
Dachang
Department of Biotechnology, Dalian Medical University, Dalian, China jimxin@hotmail.com.
Y
Y
Xin
D000818:Animals; D017963:Azithromycin / Q000737:chemistry; D001644:Bifidobacterium / Q000187:drug effects; D001835:Body Weight; D015169:Colony Count, Microbial; D058645:Denaturing Gradient Gel Electrophoresis; D004926:Escherichia coli / Q000187:drug effects; D005243:Feces / Q000382:microbiology; D000069196:Gastrointestinal Microbiome / Q000187:drug effects; D007422:Intestines / Q000382:microbiology*; D007779:Lactobacillus acidophilus / Q000378:metabolism*; D008297:Male; D016133:Polymerase Chain Reaction; D019936:Probiotics / Q000737:chemistry*; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D012336:RNA, Ribosomal, 16S / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D013211:Staphylococcus aureus / Q000187:drug effects
D012336:RNA, Ribosomal, 16S; D017963:Azithromycin