Evaluation of Probiotic Supplementation as an Intervention for Antibiotic-induced Stress in Murine Model

Abstract

Escalation of antibiotics usage in recent years has imposed serious impacts on physical as well as on mental health. Overuse of antibiotics is known to cause imbalance of gut microbiome profile which in turn can lead to the development of stress and anxiety like behaviors. However, efficacy of restoring disturbed gut microbiome after the overuse of Ciprofloxacin by using probiotic Bacillus clausii is not well researched. In this research, we developed an animal model by using 24 BALB/c male mice of weight 25 mg ± 10mg and divided them equally into two control groups; positive control and negative control, and into two experimental groups to study the effect of Ciprofloxacin and Bacillus clausii on gut microbiota and behavioral response. In negative control group, mice were provided with appropriate food and saline water for 3 weeks. In positive control group, 135mg/kg/d of antibiotic Ciprofloxacin was administered to each mice for period of 3 weeks. In the first experimental group, mice received antibiotic Ciprofloxacin dosage of 135mg/kg/d and were alongside treated with the probiotic Bacillus clausii, with a time interval of three hours between administrations for a period of 3 weeks. In experimental group 2, mice were first administered with antibiotic Ciprofloxacin for 3 weeks followed by treatment with probiotic Bacillus clausii for 1 week. After the end of treatment plan, different behavioral tests including Elevated Plus Maze test, Tail suspension test, Forced Swimming test and Marble Burrowing test were performed on mice to examine the effect of Ciprofloxacin and Bacillus clausii on neurobehavioral changes in body. Fecal samples were collected after the end of treatment period to analyze and estimate the population of Bacillus clausii in gut using Tryptic Soya Agar (TSA) microbial culture testing technique . Results of behavioral tests revealed that experimental group 1 (Ab+pb), showed the most significant results and found to be most mobile among all other groups. In experimental group 2 (Ab with pb ), behavioral response of mice was found to be less efficient, showing lesser activity, as compared to the experimental group 1 . Whereas, positive control group (Ab only) displayed the least mobility, demonstrating depressant effect of Ciprofloxacin. In negative control group (Normal saline), mice showed no anxiety and depression like behavior and their behavioral response was found to be consistent with normal Page | 13 behavior. Microbial culture test results showed that probiotic Bacillus clausii was able to sustain most efficiently in experimental group 1 (Ab+pb) in comparison with other groups. These results revealed that administration of probiotic Bacillus clausii along with antibiotic mitigated the antibiotic-induced stress. Simultaneous supplementation of probiotic Bacillus clausii along with Ciprofloxacin could alleviate the mental stress and facilitate the restoration of gut dysbiosis caused by Ciprofloxacin.