Scientific- Fame Pharmaceuticals Company

IN VITRO ANTIBACTERIAL ACTIVITY OF "DIARSTAT"
By
Dr. Khin Maung Lwin MB,BS, Ph.D., Dr. B.A.
and
Dr. Myint Oo MSc, DS, Ph.D.

FAME Pharmaceuticals Enterprise
Yangon, Myanmar

ABSTRACT

A herbal formulation "Diarstat" produced by FAME Pharmaceuticals Enterprise has been tested on Bacillus subtillis, Escherichia coli (ETEC; STLT), Pseudomonas pyocyanea, Shigella sonnei and Vibrio cholerae (091; 092;01 strains). Tetracycline HCl was used as control drug. Standard agar disc diffusion method was used through out the experiment. The zones of inhibition recorded were 14mm with B. subtillis, 14mm with E. coli (ETEC), 18mm with E. coli (STLT), 13mm with P. pyocyanea, 20mm with S. sonnei, 14mm with V. cholerae (01), 20mm with V. cholerae (091) and 14mm with V. cholerae (092).

INTRODUCTION

Although diarrheal illness is not the over whelming health problem in the developed countries, it is still account for as much as 20% of the acute care visits by young children at hospitals in developing and under developed countries (Kotloff et al., 1988). In Myanmar, diarrheal illness is one of the top ten diseases defined by Myanmar National Health Programme. In order to combat the problems of diarrhea globally, the World Health Organization in its Diarrheal Disease Control programme has given a special emphasis on the use of traditional folklore medicines in the control and management of diarrhea (Anonymous, 1979). Therefore, FAME Pharmaceuticals Enterprise has formulated a combined herbal extract preparartion called "Diarstat" with six Myanmar medicinal plant products, namely Acacia catechu, Eugenia jambolana, Ferula foetida, Leucaena glauca, Quisqualis indica, and Zingiber officinale.

In the present paper, we have described the in vitro antibacterial activity of "Diarstat" on some of the pathogenic bacteria viz. Bacillus subtillis, Escherichia coli (ETEC; STLT), Pseudomonas pyocyanea, Shigella sonnei and Vibrio cholerae (091; 092;01 strains).

MATERIALS AND METHODS

Preparation of "Diarstat": The formulation of "Diarstat" contains the extracts of Acacia catechu, Eugenia jambolana , Ferula foetida, Leucaena glauca, Quisqualis indica and Zingiber officinale in the ratio of (6:38:6:38:140:6). The combined extract was dissolved in 1% DMSO and the dilutions were made with Phosphate Buffered Saline (PBS) pH 7.2 . The test drug concentration was 100 µg/ml.

Screening of in vitro antibacterial activity: Screening of antibacterial activity was done by impregnated filter-paper discs. The discs, 8mm in diameter, and punched from No. 1 Whatman filter paper was sterilized by autoclaving followed by drying in oven. It was then impregnated with diluted extract and then allowed to dry at room temperature under sterile condition. Test organisms used were Bacillus subtillis, Escherichia coli (ETEC; STLT), Pseudomonas pyocyanea, Shigella sonnei and Vibrio cholerae (091; 092;01 strains). A few colonies of the pure cultures were inoculated into 2-4ml of trypticase soy broth. These tubes were incubated for 2-4 hours in a water bath at 37^oC to produce a bacterial suspension with barely visible to moderate cloudiness. This contained approximately 10⁵ or 10⁷ organisms per ml. The organisms were then streaked onto surface of Oxoid Sensitivity test agar with a swab. After the inoculum had dried, the dried discs were placed on the medium. A control disc (tetracycline) was also placed onto the medium. After overnight incubation at 37^oC, the zones of inhibition were measured.

RESULTS

Figures 1,2,3,4,5 and 6 represents the antibacterial activity of "Diarstat" against Bacillus subtillis, Escherichia coli (ETEC; STLT), Pseudomonas pyocyanea, Shigella sonnei and Vibrio cholerae (091; 092;01 strains). The sensitivity of "Diarstat" was compared with tetracycline by measuring the zones of inhibition. "Diarstat" was found to be antibacterial against Bacillus subtillis, Escherichia coli (ETEC; STLT), Shigella sonnei and Vibrio cholerae (091; 092;01 strains) with the mean diameter of >14mm. "Diarstat" was not very sensitive against Pseudomonas pyocyanea (zone of inhibition 13mm).

DISCUSSION

In subjects with diarrhea, fluid and electrolytes replacement are the most popular treatment. Fluid therapy often continues to consist of clear liquid such as fruit juice and carbonated beverages, that are not suitable for rehydration therapy because of their high osmolarity or inappropriate ratio of sodium to carbohydrate (Snyder, 1991). Oral rehydration therapy (ORT) is the replacement therapy for intravenous dextrose-saline infusion. If patients with diarrhea is often accompanied by bacterial infection, antibiotic therapy is considered in order to shorten the course of illness. In traditional medicine system, many plants or herbs are claimed to have anti-diarrhoeal efficacy without any scientific basis. The aim of the present study was to evaluate the antibacterial activity of Myanmar reputed traditional formulation "Diarstat" with particular reference to pathogenic bacteria potential to cause diarrhea.

CONCLUSION

It is concluded that "Diarstat" showed in vitro antibacterial activity against some pathogenic bacteria causing diarrhea. Further studies with in vivo animal model and clinical trials should be carried out.