THE EFFICACY OF TSAANG GUBAT ( Ehretia microphylla Lam ) DECOCTION AS AN ANTI-DIARRHEAL AGENT VERSUS ATROPINE SULFATE GR

1. THE EFFICACY OF TSAANG GUBAT (Ehretia microphylla Lam) DECOCTION AS AN ANTI-DIARRHEAL AGENT VERSUS ATROPINE SULFATEGROUP 2A BUDAO, Cherry Pinky M. BAŇAREZ, Karla Kristel A. BAQUIRAN, Jessica Aevan U. BASMAYOR, Edwin Marlon C. BASTE, Charisse Liz P. BAUTISTA, Jose Antonio L. BEESLA, Sundeep Kaur B. BELMONTE, Carlo Alphonso BONDOC, Hidelisa E. BORROMEO, Christian Leo P. BRION, Marco Alberto C. BUENSALIDA, Rainier John BULAONG, Marie Veronica G. BUMANGLAG, Niña M. BURGO, Terence Aaron L. BUŇAG, Mark Chester Victor T. CACDAC, Maybelle Chris CAMACLANG, Marie Len A. CANADALLA, Kristine Joyce L. CAOILI, Sylvia Nica J. CASTILLO, Maria Carmella P. CHAN, Jose Carlos A. CHAVEZ, Frances Joy T.

2. INTRODUCTION

3. ABSTRACT • Diarrhea • ↑ stool mass, ↑ stool frequency, ↑ stool fluidity • > 200 gm/da • severe: > 14 L/da • w/o fluid resuscitation: death • increase in stool features • acute: > 2 wks • persistent: 2-4 wks • chronic: > 4 wks

4. Agents / Factors: Escherichia coli Campylobacter jejuni Shigella spp. Aeromonas spp. side effects from medications i.e. antibiotics, bronchodilators, antacids, laxatives assoc DO: lactose intolerance, IBS PATHOGENESIS

5. Other Assoc'ns ingestion of poisonous substances e.g. organophosphate insecticides amanita and other mushrooms arsenic cpds preformed environmental toxins PATHOGENESIS

6. APPROACH TO DIARRHEA fluid and electrolyte replacement oral rehydration solutions : replace lost fluids antibiotics : elimination of any causative agent elimination of dietary lactose : suppressing the underlying mechanism mild opiate i.e. Loperamide® : cases of mild to moderate diarrhea

7. LOCAL HERBAL MEDICATIONS primary health care increasing cost found to be effective in tx of common ailments attested by the NSDB advocated by the DOH “Tsaang Gubat” Ehretia microphylla Lam. tx: diarrhea, stomachache

8. BACKGROUND OF THE STUDY to determine the efficacy of “Tsaang Gubat” as an alternative anti-diarrheal drug tested along with Atropine Sulfate and Normal Saline Solution “Tsaang Gubat” as effective as Atropine, as a very good alt drug for diarrheal cases in poverty-stricken communities due to lack of funds for purchasing expensive anti-diarrheal drugs

9. STATEMENT OF THE PROBLEM • Philippines is the second largest contributor to diarrhea morbidity in the world, next to China. • DOH and UNICEF: • over 70,000 Filipino children die • span of 7 years • WHO: • fourth leading cause of death among children ≤ 4 y.o. • third leading cause of death among Filipino children • Philippines ranks second among 13 countries with most number of diarrhea cases

10. OBJECTIVE to determine the effectiveness of Tsaang Gubat, one of the 10 herbal medicines launched by the DOH as an anti-diarrheal medication, compared to the standard drug, Atropine By using the test drug Tsaang Gubat, was the distance travelled by the charcoal meal shortened? Is Tsaang Gubat for treatment of diarrhea? Is Tsaang Gubat as effective as the standard drug, Atropine, for the treatment of diarrhea?

11. SIGNIFICANCE OF THE STUDY • trend of utilizing medicinal plants from treating common ailments (e.g. colds) to extreme cases (e.g. cancer) • more scientific tests conducted to verify the efficacy • commonly advocated by our forefathers and our neighborhood albularios • government introduced several programs aimed at promoting and further developing this to bolster the health care sector

12. SIGNIFICANCE OF THE STUDY • Tsaang Gubat • one of the herbal medicines listed in “Sampung Halamang Gamot” program in the early 1990's • shrub commonly found in the Philippines • endorsed as anti-spasmodic for abdominal pains and for other GI DO (diarrhea, dysentery) • listed in the BFAD as medicinal plant

13. SCOPE AND LIMITATION • limited to the effects of the medicinal plant in decreasing intestinal motility = beneficial of diarrhea or LBM • no other forms of treatment for diarrhea or LBM have been included • Normal Saline Solution has no known effect on GI motility • Atropine is a known muscarinic (cholinergic) blocker = inhibits GI motility

14. SCOPE AND LIMITATION (con't) starved mice of the same sex and about the same weight as subjects = prevent bias d/t diff in sex and wt starvation so as not to affect drug absorption and prevent any obstruction in the GIT for the charcoal sol'n

15. SCHEMATIC DIAGRAM

16. METHODOLOGY % distance travelled as indicator of anti-diarrheal activity parameter measured is the length of the small intestine travelled by the charcoal

17. METHODOLOGY • Male and female albino mice obtained from UERM • day prior to the expt, mice were placed in wire meshed cages, given standard pellet diet and water • experimentation carried out accdg to the IAEC guidelines • mice were weighed and labeled for proper ID • divided into three groups: • Positive Control (Atropine Sulfate) • Negative Control (Normal Saline Solution) • Test Drug (“Tsaang Gubat”)

18. METHODOLOGY (con't) • 9 mice in total; 3 mice per group • Negative control: Normal Saline Solution via gavage 0.5 mL/kg orally • Positive control: Atropine Sulfate via gavage 10mg/kg • Test drug: Ehretia microphylla Lam. (Tsaang Gubat”) extract via gavage 10g/kg orally

19. METHODOLOGY (con't) Charcoal sol'n 10 gm charcoal mixed with 100 mL castor oil 20 mL of charcoal-castol oil suspension then mixed with 10 mL coconut oil suspension stirred constantly to obtain uniform suspension

20. METHOLODOGY (con't) 2 kg dried leaves of “Tsaang Gubat” chopped into small pieces, consequently boiled in 1L of distilled water for 8 hrs boiled after filtering using filter paper, collected in a beaker repeated twice until dark-brown extract was obtained refrigerated for 1 hr prior to the expt

21. METHODOLOGY • ANIMATION OF METHODOLOGY

22. COLLECTION OF DATA 20 mins after, mice were sacrificed intestines were excised (from pylorum to cecum) distance travelled by activated charcoal measured and recorded corresponding to the different doses administered total length was also measured Percentage Distance = Activated charcoal___ X 100 Total length of intestine

23. ANALYSIS OF DATA weight of the rat dose of drug administered length of intestine charcoal distance length of the small intestine travelled by the charcoal percent of distance travelled = Length travelled by activated charcoal x 100 total length of intestine

24. RESULTS AND DISCUSSION

26. RESULTS AND DISCUSSION Table 1.0. Mean and Standard Deviation of the Percentage of Distance Travelled by Charcoal with administration of Atropine and Normal Saline Solution Distance Travelled by Charcoal

27. RESULTS AND DISCUSSION Table 2.0. T-test Values for the Individual Samples in the Positive Control Group and the Negative Control Group

29. RESULTS AND DISCUSSION Table 3.0. Mean and Standard Deviation of the Percentage of Distance Travelled by Charcoal with administration of Tsaang Gubat and Normal Saline Solution Distance Travelled by Charcoal

31. Distance Travelled by Charcoal

33. RESULTS AND DISCUSSION • Mouse 1: 10 cm distance (27% of the total intestinal length of 36 cm) • Mouse 2: 12 cm distance (30% of the total intestinal length of 40 cm) • Mouse 3: 18 cm distance (50% of the total intestinal length of 36 cm)

34. RESULTS AND DISCUSSION

35. CONCLUSION • Negative Ctrl Grp (NSS) • isotonic solution; same conc as body cells; N physiologic conditions in the intestine • ACTUAL: shorter % distance travelled in the intestine • THEORETICAL: longer % distance travelled in the intestine

36. Atropine Sulfate antimuscarinic / anticholinergic agent competitive antagonist of Ach on the muscarinic receptor ↓ GI motility & secretion ACTUAL: longer % distance travelled in the intestines THEORETICAL: shorter % distance travelled in the intestine

37. RECOMMENDATIONS

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