THE EFFICACY OF TSAANG GUBAT (Ehretia microphylla Lam) DECOCTION AS AN ANTI-DIARRHEAL AGENT VERSUS ATROPINE SULFATE GROUP 2A

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

GROUP 2A

BUDAO, Cherry Pinky M.BAAREZ, 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 AlphonsoBONDOC, Hidelisa E.BORROMEO, Christian Leo P.BRION, Marco Alberto C.BUENSALIDA, Rainier John BULAONG, Marie Veronica G.BUMANGLAG, Nia M.BURGO, Terence Aaron L.BUAG, Mark Chester Victor T.CACDAC, Maybelle ChrisCAMACLANG, Marie Len A.CANADALLA, Kristine Joyce L.CAOILI, Sylvia Nica J.CASTILLO, Maria Carmella P.CHAN, Jose Carlos A.CHAVEZ, Frances Joy T.1INTRODUCTION2Agents / Factors:Escherichia coliCampylobacter jejuniShigella spp.Aeromonas spp.side effects from medications i.e. antibiotics, bronchodilators, antacids, laxativesassoc DO: lactose intolerance, IBSPATHOGENESIS4Diarrhea is caused by several agents or factors, such as infectious agents which include E. Coli, Campylobacter, Shigella, Aeronomas, and Salmonella. Other factors include side effects from medications such as antibiotics, bronchodilators, antacids, and laxatives. Disorders associated with diarrhea include lactose intolerance and irritable bowel syndrome.Other Assoc'nsingestion of poisonous substances e.g. organophosphate insecticidesamanita and other mushroomsarsenic cpdspreformed environmental toxinsPATHOGENESIS5Diarrhea is also associated with ingestion of poisonous substances such as organophosphate insecticides, amanita and other mushrooms, arsenic, and pre-formed environmental toxins.APPROACH TO DIARRHEAfluid and electrolyte replacementoral rehydration solutions : replace lost fluidsantibiotics : elimination of any causative agentelimination of dietary lactose : suppressing the underlying mechanismmild opiate i.e. Loperamide : cases of mild to moderate diarrhea6An approach to diarrhea is usually directed towards fluid and electrolyte replacement. Oral rehydration solutions are given to replace lost fluids. Other management solution for diarrhea is elimination of any causative agent involved by giving antibiotics. In other cases which involve other underlying disease states, management options are focused on suppressing the underlying mechanism, such as elimination of dietary lactose in lactose-intolerant persons. A mild opiate such as Loperamide may be effective in cases of mild to moderate watery diarrhea.LOCAL HERBAL MEDICATIONSprimary health careincreasing costfound to be effective in tx of common ailments attested by the NSDBadvocated by the DOHTsaang GubatEhretia microphylla Lam.tx: diarrhea, stomachache7As part of primary health care and because of the increasing cost of drugs, the use of locally available medicinal plants has been advocated by the Department of Health. Many local plants and herbs in the Philippine backyard and field have been found to be effective in the treatment of common ailments as attested by the National Science Development Board, other government and private agencies/persons engaged in research. The Department of Health is advocating the use of Tsaang Gubat (Ehretia microphylla Lam.) for diarrhea and stomachache.BACKGROUND OF THE STUDYto determine the efficacy of Tsaang Gubat as an alternative anti-diarrheal drugtested along with Atropine Sulfate and Normal Saline SolutionTsaang 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 drugs8The study was conducted to determine the efficacy of Tsaang Gubat as an alternative anti-diarrheal drug. Compared with Atropine and Normal Saline Solution, the effectiveness of the said herbal medication was tested along with the other drugs. The researchers believe that the efficacy of Tsaang Gubat is as great as that of Atropine and, if proven so, can be used as a very good alternative drug for diarrheal cases which is predominant in poverty-stricken communities which lack funds for purchasing expensive drugs to treat diarrheal casesOBJECTIVEto 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, AtropineBy 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?10The purpose of the study is to compare the anti-diarrheal efficacy of Tsaang Gubat, of the 10 herbal medicines launched by the Department of Health in early 1990s, to the standard drug Atropine.

The purpose of the study is to answer the following questions:1) By using the test drug Tsaang Gubat, wasthe distance travelled by the charcoal meal shortened?2) Is Tsaang Gubat for treatment of diarrhea?3) Is Tsaang Gubat as effective as the standard drug, Atropine, for the treatment of diarrhea?SIGNIFICANCE OF THE STUDYTsaang Gubatone of the herbal medicines listed in Sampung Halamang Gamot program in the early 1990'sshrub commonly found in the Philippinesendorsed as anti-spasmodic for abdominal pains and for other GI DO (diarrhea, dysentery)listed in the BFAD as medicinal plant12Tsaang Gubat is a shrub that is found abundantly in the Philippines. It is endorsed by the DOH as an antispasmodic for abdominal pains. It is also used for other gastrointestinal disorders like diarrhea and dysentery. Researches and tests conducted on Tsaang Gubat have proven its worth as herbal medicine as such, aside from being endorsed by the DOH, it is also listed as a medicinal plant with the Bureau of Food and Drugs (BFAD). Thus, studies on herbal medicines like Tsaang Gubat must continue as these can provide cheap, locally available and easily accessible alternative medicines for many ailments and other health conditions.SCOPE AND LIMITATIONlimited to the effects of the medicinal plant in decreasing intestinal motility = beneficial of diarrhea or LBMno other forms of treatment for diarrhea or LBM have been includedNormal Saline Solution has no known effect on GI motilityAtropine is a known muscarinic (cholinergic) blocker = inhibits GI motility13Although the review of related literature presents a number of indications for the use of tsaang gubat, the present study was only limited to the effects on the medicinal plant in decreasing intestinal motility which is beneficial of diarrhea or loose bowel movement. Furthermore, no other forms of known treatment for diarrhea or loose bowel movement have been included in the study. Atropine sulfate was used as the positive control and Normal Saline Solution as negative control. The NSS has no known effect on gastrointestinal motility while atropine on the other hand, is a known muscarinic blocker that inhibits intestinal motility.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 wtstarvation so as not to affect drug absorption and prevent any obstruction in the GIT for the charcoal sol'n14The subjects used in the experiment were starved mice of the same sex and have about the same weight. This was done to prevent bias due to differences in sex and weight, the mice had been starved so as not to affect the absorption of drugs and to prevent any obstruction in the GI tract for the way of the charcoal solution. SCHEMATIC DIAGRAM15METHODOLOGY% distance travelled as indicator of anti-diarrheal activityparameter measured is the length of the small intestine travelled by the charcoal16METHODOLOGYMale and female albino mice obtained from UERMday prior to the expt, mice were placed in wire meshed cages, given standard pellet diet and waterexperimentation carried out accdg to the IAEC guidelinesmice were weighed and labeled for proper IDdivided into three groups:Positive Control (Atropine Sulfate)Negative Control (Normal Saline Solution)Test Drug (Tsaang Gubat)17Albino mice (Male/Female) obtained from the UERM technician were used for the experiment. A day prior to the experiment, the mice were placed in cages that are wire meshed and were given with standard pellet diet and water. The mice were subjected to fasting 12 hours prior to the experiment. All the animal experimentation in this study was carried out according to the guidelines of Institutional Animal Ethics Committee (IAEC). The mice were weighted and labelled for proper identification. The mice were divided into three groups for administration of Positive control, Negative control, and Test drug.METHODOLOGY (con't)9 mice in total; 3 mice per groupNegative control: Normal Saline Solution via gavage 0.5 mL/kg orallyPositive control: Atropine Sulfate via gavage 10mg/kgTest drug: Ehretia microphylla Lam. (Tsaang Gubat) extract via gavage 10g/kg orally18Nine mice were divided to such groups. The negative control group received Normal Saline Solution via gavage with the dose of 0.5 ml/kg orally. Positive control group received atropine sulfate via gavage with the dose of 10 mg/kg orally. Test group had received via gavage Ehretia microphylla Lamextract with the dose of 10 g/kg orally. METHODOLOGY (con't)Charcoal sol'n10 gm charcoal mixed with 100 mL castor oil20 mL of charcoal-castol oil suspension then mixed with 10 mL coconut oilsuspension stirred constantly to obtain uniform suspension19Charcoal solution was prepared as follows: 10 g of charcoal was mixed with 100 ml castor oil. 20 ml from this suspension was then mixed with 10 ml coconut oil. The suspension was stirred constantly to obtain a uniform suspension.METHOLODOGY (con't)2 kg dried leaves of Tsaang Gubat chopped into small pieces, consequently boiled in 1L of distilled water for 8 hrsboiled after filtering using filter paper, collected in a beakerrepeated twice until dark-brown extract was obtainedrefrigerated for 1 hr prior to the expt20 Two kilograms of Tsaang Gubat (Ehretia microphylla Lam) dried leaves were chopped into small pieces, and consequently boiled in one litter of distilled water for 8 hours. It was boiled after filtering using filter paper and was collected in a beaker. The process was repeated two times until a dark-brown extract was obtained. The extract collected in a beaker was refrigerated for 1 hour prior to the experiment.METHODOLOGYANIMATION OF METHODOLOGY21COLLECTION OF DATA20 mins after, mice were sacrificedintestines were excised (from pylorum to cecum)distance travelled by activated charcoal measured and recorded corresponding to the different doses administeredtotal length was also measuredPercentage Distance = Activated charcoal___ X 100 Total length of intestine22Twenty minutes after charcoal intake, mice were sacrificed. Then intestines were excised from pylorus to cecum. The distance travelled by the activated charcoal was measured and recorded corresponding to the different doses administered. The total length of the intestine was also measured. Mice were dispensed appropriately. Percentage of distance travelled was calculated using the formulaANALYSIS OF DATAweight of the ratdose of drug administeredlength of intestinecharcoal distancelength of the small intestine travelled by the charcoal

percent of distance travelled= Length travelled by activated charcoal x 100 total length of intestine23The parameter to be measured in the experiment is the length of the small intestine traveled by the charcoal. The data on weight of the rat, dose of drug administered, length of intestine, charcoal distance and percent of distance traveled by the charcoal were obtained from all groups. Percentage of distance traveled was obtained by using the formula: (Length traveled by activated charcoal total length of intestine) x 100.RESULTS AND DISCUSSION2425Independent T-Test was performed to evaluate significant difference in the mean percentages of distance travelled by the charcoal in the intestine of the mice given with Atropine (positive control) and Normal Saline solution (negative control). RESULTS AND DISCUSSION

Table 1.0. Mean and Standard Deviation of the Percentage of DistanceTravelled by Charcoal with administration of Atropine and NormalSaline SolutionDistance Travelled by Charcoal26Table 1.0 gives the descriptive statistics for each of the two groups. There are 18 experimental mice for each drug group with an average of 23.2267 in the effect of the drug which is the percentage of distance travelled by the charcoal for the positive control group using atropine, with a standard deviation of 23.85. For the negative control group administered with NSS, the mean of percentage of distance travelled is 46.65 with a standard deviation of 22.05.

RESULTS AND DISCUSSIONTable 2.0. T-test Values for the Individual Samples in the Positive ControlGroup and the Negative Control Group27In Table 2.0., it can be seen that an equal variances t-test failed to reveal a statistically reliable difference between the mean percentage distance travelled between the positive control group (M = 23.23, s = 23.85) and the negative control group (M = 46.65, s = 22.05), t(34) = 3.06, since the p= .621 is higher than the p-value at level of significance which is .05. This implies that there is no significant evidence that the positive control group administered with atropine has a greater effect compared to that of the negative control group.RESULTS AND DISCUSSIONTable 3.0. Mean and Standard Deviation of the Percentage of DistanceTravelled by Charcoal with administration of Tsaang Gubat andNormal Saline Solution

Distance Travelled by Charcoal29The mean percentage of distance travelled by the charcoal in the intestines of the mice given with Tsaang Gubat wild Tea (Ehretia microphylla Lam) (test group) against the negative control (normal saline solution) is presented in Table 3.0. It can be seen that in the test group, the mean distance travelled by the tsaang gubat decoction was 31.35 with a mean standard deviation of 15.68 while the negative control (NSS) has a mean distance travelled by 46.6561 with a mean standard deviation of 22.05.RESULTS AND DISCUSSIONMouse 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)33Based on the groups experimental results in the negative control group, 0.5 ml of Normal saline was administered to three mice. The charcoal tracing of Mouse 1 traveled a distance of 10 cm, which is 27.7% of the total intestinal length of 36 cm.

The charcoal tracing of Mouse 2 traveled a distance of 12 cm, which is 30% of its total intestinal length of 40 cm. While the charcoal tracing of Mouse 3 traveled a distance of 18 cm, which is 50% of its total intestinal length of 36 cm.

The groups experimental results showed that Mouse 3 of the negative control group covered a longer travel distance of charcoal as compared to Mouse 1 and Mouse 2 of the same group. Whereas, mouse 1 and mouse 2 of the negative control group covered a shorter travel distance as compared to the mouse 1 and mouse 2 of the positive control group. Mouse 1 of the positive control group showed the greatest distance traveled as compared to both Positive and Negative control groups.RESULTS AND DISCUSSION

34CONCLUSIONNegative Ctrl Grp (NSS)isotonic solution; same conc as body cells; N physiologic conditions in the intestineACTUAL: shorter % distance travelled in the intestineTHEORETICAL: longer % distance travelled in the intestine35In general, the negative control group (normal saline) showed a shorter distance traveled in the intestines in terms of percentage as compare to the positive control group (Atropine sulfate).

The Normal Saline Solution administered to the Negative control group theoretically should have shown a longer travel distance of the charcoal in the intestine as compared to the positive control group, which is administered with Atropine Sulfate.

RECOMMENDATIONS37REFERENCESAlcantara, A. (2000). Community Health Nursing Services in the Philippines. (9th ed.). PhilippinesAlvarez, A. A. (2010, January 18).Tsaang Gubat/Wild Tea (Ehretia microphylla Lam.). Retrieved August 22, 2010 from http://www.philippineherbalmedicine.org/tsaang_gubat.htmBabu, R. D. S., et al. (2009). Antidiarrheal Activity of Cynodon Dactylon. Pers. Pharmacognosy Magazine, 5(19), 23-7.Cinco, E. B. (2010, June 13). Alternative Health Care. Retrieved August 20, 2010 from http://www.mb.com.ph/node/261899Cortez-Maramba, N., et al. (1983). Guidebook on the Proper Use of Medicinal Plants. National Science and Technology Authority-University of the Philippines. Quezon City: Katha Publishing Co. Inc.Das, A. K., et al. (2009). Evaluation of Anti-diarrhea Activity of Rhizophora mucronata Bark Extracts. The Internet Journal of Alternative Medicine. ISSN: 1540-2584.Fauci, A., Braunwald, E., Kasper, D., Hauser, S., Longo, D., Loscalzo, J. (2008). Harrison's Principles of Internal Medicine (17th ed.). United States of America: The McGraw-Hill Companies, Inc.GMA News TV. Diarrhea kills 10,000 Pinoy kids every year WHO. (2008, September 12). Retrieved August 22, 2010 from http://www.gmanews.tv/story/119921/Diarrhea-kills-10000-Pinoy-kids-every-year---WHOKatzung, B. G., Masters, S. B., & Trevor, A. J. (2009). Basic and Clinical Pharmacology (11th ed.). Singapore: McGraw Hill Companies, Inc.38