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Investigatory Project

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Introduction
The investigatory project designs to help indirectly the common people on the use of indigenous floral resources such as Alstonia Scholaris that serves as a material to regulate and inhibit biotic potential of the individual.

1.1 Background of the Study
The country’s current population is estimated at 93 million people (NSO Annual Report: 2010) where about 65 percent are living in urban area and experience poverty due to the usual family practice of giving birth. Pro-RH Bill lawmakers have presented the effective measures to control the growing population and provided alternative ways to maintain reproductive health and plausible actions to that program. Amidst all the programs, poor families are seen and cannot sustain the basic necessities of their children and depriving them from enjoying the fullness what is due to for the kids. Young women whose ages range from 14-25 often experience fast hormonal secretions that usually associated with active and rapid biotic potential. Early sexual maturity becomes a primordial factor for fertility and early teenage pregnancy. This then is the scenario that strive the researchers to conduct experiment that provide information on the use of Alstonia Scholaris as a viable material for controlling population.

1.2 Statement of the Problem
The project undertaken by the researchers on the Alstonia Scholaris extract as applied to “rabbit” sought to answer the following questions: 1. What is the effect of Alstonia scholaris extract on the biotic potential of “Oryctolagus Cuniculus”?

2. How effective is the use of Alstonia Scholaris extract to the life of “Oryctolagus Cuniculus” in terms of:
a. antifertility
b. implantation/inhibitory of hormonal activity
c. gestation 3. To what extent is the use of Alstonia Scholaris extract before and after mating of the “Oryctolagus Cuniculus” in terms of:
a. antifertility
b. implantation/inhibitory of hormonal activity
c. gestation

1.3 Objectives
The investigatory project entitled: “Inhibitory Mechanism of Alstonia Scholaris On the Reproductive Potential of Oryctolagus Cuniculus” was conducted by the proponents to develop alternative means in controlling population. Specifically, the project was undertaken to:
1. Prove that Alstonia Scholaris is effective way in inhibiting reproductive process and cycles.
2. Make use of Alstonia scholaris as indigenous, endemic floral materials that are potential hormonal switcher for birth control. 3. compare and contrast on the effectiveness of commercially provide pill from the natural Alstonia Scholaris bark extract.

1.4 Null Hypothesis
The researchers assumed the following null hypothesis:
1. Alstonia scholaris extract is not effective inhibiting material for biotic reproduction of “Oryctolagus Cuniculus”.
2. Astonia scholaris extract does not affect the fertility, implantation, hormonal activity and gestation of “Oryctolagus Cuniculus”
3. There is no significant difference on the use of Alstonia Scholaris and commercially-made Trust Pill in terms of reproductive indices.

1.5 Significance of the Study
Alstonia Scholaris as an indigenous tree was found to have medicinal value on the fertility and inhibition of biotic potential of “Oryctolagus Cuniculus”. The researchers conducted the effectiveness of Alstonia as potential hormonal inducer, by speeding up the reproductive cycle of “Oryctolagus Cuniculus”, thus inhibiting the pregnancy of the rabbit.
Ordinary Filipinos who had been suffered from menstruation and reproductive ovulation will benefit from Alstonia for it enhances rapid discharge of undeveloped / unfertilized eggs. Alstonia Scholaris affected fertility, implantation and gestation processes occurring in the female rabbit.
Large size families whose parents were biologically potent may hinder continuous unwanted pregnancies using Alstonia bark extract. Commercially produced hormonal pill would compete medically with Alstonia Scholaris since most of the pills had increased its price in the market. Alstonia Scholaris as a viable antifertility agent helped the Filipino women through inducing hormonal secretion without jeopardizing the reproductive health and pre-natal development.
Health experts were benefitted from Alstonia.Scholaris for they recommend local plant source used as practical way to regulate unwanted pregnancy which found to be affordable and available in the country.

1.6 Scope and Limitations of the Study
The investigatory project dealt with the effect of Alstonia Scholaris, a plant belonging to Apocynaceae to the biotic potential of “rabbit”.
Materials were basically Alstonia Scholaris bark extract as employed to the sexually-matured “rabbit” with different parameters: pre-coital and post-coital conditions.
Biotic potential of the “Oryctolagus Cuniculus” was indicated by fertility, implantation and gestation at considerable number of female rabbit. Inferential statistics were employed to see the veracity of hypothetical statements concerning its inhibitory effect on sexual reproduction.
The study covered the effects of Alstonia Scholaris extract on reproductive indices and uterus examination of the rabbit. Histological examination on the ovaries for oogenesis was conducted using the treatments: positive control (Trust Pill), negative control (water) and experimental treated with Alstonia Scholaris methanolic bark extract.
The study was limited to the number of “Oryctolagus Cuniculus” about 2 rabbits subjected for experimentation. The food served to the specimen were mainly pellets and unlimited to water being served.

Review of Related Literature Plants have been used since the beginning of time for gynecological and obstetric conditions (Miller and Murray: 1998). In remote areas, traditional contraception practices still persist until today. One of the plants made use to satisfy reproductive health care by the traditional birth attendants is Alstonia Scholaris known locally as “dita” or “bita”. Decoction of the bark of Alstonia scholaris has been used indigenously to induce menstruation or to prevent rapid pregnancy. Dita is a smooth tree growing 6 to 20 meters high. Branches are lenticellate. Bark is dark greyish, somewhat rough, yielding an abundant, bitter, and milky sap. Leaves are in whorls,4 to 7in a whorl, leathery, narrowly obovate to spatulate, 10 to 20 centimeters long, 3 to 4.5 centimeters wide, pointed at the base, rounded at the apex, glossy green on the upper surface, white or grayish on the underside. Laternal nerves are very numerous, parallel, and terminating in a intramarginal vein. Flowers are crowded, numerous, somewhat hairy, greenish-white, about 1 centimeter long, hairy in the throat, borne in compact, hairy cymes about 10 centimeters long. Fruits are made up of two slender follicles which are pendulous and cylindric follicles, 20 to 40 centimeters long, 4-5 millimeters in diameter. Seeds are 3 to 4 millimeters long, with brown ciliate hairs on the ends. The bark is almost odourless and very bitter, with abundant bitter and milky sap. Alstonia Scholaris has many uses an reported as antimicrobial, antiamoebic, antidiarrheal, antihypertensive, antimalarial, febrifuge, stimulant, hepatoprotective, immunomodulatory, anti-cancer, antiasthmatic, antioxidant, analgesic, antiinflammatory, anti-fertility, anti-diabetic, cardiotonic.
The bark contains the alkaloids ditamine, crystalline and toxic echitamine; uncrystallizable and bitter principle, and echitamine that is used to serve as an alternative to quinine. Extracts prepared from the plant has been reported to possess cytotoxic activity. At one time, a decoction of the bark was used to treat diarrhoea and malaria, as a tonic, febrifuge, emmenagogue, anticholeric and vulnerary. Decoctions of the leaves were used for beriberi. Ayurveda recommends Alstonia Scholaris for bowel complaints.
An ethanol extract of the bark of Alstonia scholaris enhanced the anticancer activity of berberine in the Ehrlich ascites carcinoma-bearing mice. This extract also showed cytotoxic activity to Hela cells. It contains echitamine and loganin as major compounds and could potentially be used as an anti-irritation agent. In India the bark of Alstonia Scholaris is used solely for medicinal purposes, ranging from Malaria and epilepsy to skin conditions and asthma. In Ayurveda it is used as a bitter and as an astringent herb for treating skin disorders, malarial fever, urticaria, chronic dysentery, diarrhea, in snake bite and for upper purification process of Panchakarma .The Milky juice of the tree is applied to ulcers.
The study made by Dr. Gupta RS, Sharma R, Sharma A, Bhatnager AK, Dobhal MP, Joshi YC and Sharma MC (2002) proves antifertility effect of Alstonia Scholaris bark extract in male rats because they conducted an experiment. They use male Wistar rats to test the effectiveness of Alstonia scholaris bark extract and was given by oral route at a dose of 200 mg/day for 60 days. The fertility and testicular function were assessed by mating tests, sperm motility, sperm concentration, biochemical indices and testicular cell population dynamics. Oral feeding with the extract at a dose of 200 mg/day for the period of 60 days did not cause body weight loss, while the weights of testes, epididymides, seminal vesicle and ventral prostate were significantly reduced. The production of step-19 spermatids was reduced by 79.6 % in treated rats. The populations of preleptotene and pachytene spermatocytes were decreased by 61.9 % and 60.1 %, respectively. Spermatogonia and Sertoli cell population were also affected. The seminiferous tubule and Leydig cell nuclear area were reduced significantly when compared to the controls. Reduced sperm count and motility resulted in a total suppression of fertility. A significant fall in the protein and sialic acid content of the testes, epididymides, seminal vesicle and ventral prostate as well as glycogen content of testes were also noticed. The fructose content in the seminal vesicle was lowered whereas the testicular cholesterol was elevated as compared with the controls. The following compounds were obtained from the extract with chromatographic separation over Si-gel column: a-amyrin, b-amyrin, lupiol acetate, venenative, rhazine and yohimbine. At the dose level employed, Alstonia scholaris bark extract has a significant antifertility effect in male rats; the primary site of action may be post meiotic germ cells.
They also have a study that was undertaken to evaluate the antifertility activity of the active principle, i.e. lupeol acetate, isolated from benzene extract of Alstonia Scholaris in other species of male rats that was the albino rats. The treatment with lupeol acetateat the dose level of 10 mg/rat/day did not cause any significant change in the body weights, but significant reduction in the weight of reproductive organs, i.e. testes, epididymides, seminal vesicle and ventral prostate, was observed. Testicular sperm count, epididymal sperm count and motility were found significantly declined when compared with controls, which resulted in reduction of male fertility by 100%. Arrest of spermatogenesis was noted at various stages with production of primary spermatocytes (preleptotene and pachytene), secondary spermatocytes and step-19 spermatids were decreased by 52.36, 54.91, 55.67 and 69.65%, respectively. The seminiferous tubules appeared reduced in size by 24.62%. Cross-sectional surface area of Sertoli cells as well as their counts was found to be significantly depleted. Leydig cell nuclear area and number of mature Leydig cells were decreased by 27.65 and 35.47%. Biochemical parameters of tissues i.e. protein, sialic acid, glycogen and cholesterol content of testes and seminal vesicular fructose also showed significant reduction. In the present study, the antifertility activity of Alstonia Scholaris bark methanolic extract was investigated in female mice. This would validate its use in folk medicine since so far no attempts have been done to popularize locally grown plants as contraceptives. Methodology
Materials and Methods
1. Collection and Extraction Stem barks of Alstonia scholaris were collected from San Francisco Compound, Quezon City, dried in a cabinet drier at 40-50 degrees Celsius homogenized in a blender and cooked in methanol (2g. of dried plant material per 5ml of methanol) for 48 hrs. with constant shaking. The methanolic extract were filtered with Whatman filter paper no.42 and where concentrated in vacuo at 45 degrees Celsius using rotary evaporator (Heidolph: WB 2000). The methanolic extracts of the stem bark of the plant were used to test its biological activity.

2. Experimental Animal

Domesticated rabbits (Oryctolagus cuniculus) were used. They were housed in a cage that is five times as large as them and maintained under natural conditions in a covered shed. Rabbits were fed with Timothy-based pellets and tap water. An initial of 5 pairs of rabbits were raised to produce offspring for two generations. The second generation offspring were used in the experiment. The offspring were raised until sexual maturity and allowed to give birth once to show that they were of proven fertility before use in the experiment…...

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