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Chapter I

THE PROBLEM

Introduction

Medicinal plants have been identified and used throughout human history. The

term medicinal plant include various types of plants used in herbalism ("herbology" or

"herbal medicine"). It is the use of plants for medicinal purposes, and the study of such

uses. The word herb has been derived from the Latin word, herba and an old French

word herbe. Nowadays, herb refers to any part of the plant like fruit, seed, stem, bark,

flower, leaf, stigma or a root, as well as a non-woody plant. Earlier, the term herb was

only applied to non-woody plants, including those that come from trees and shrubs. These

medicinal plants are also used as food, flavonoid, medicine or perfume and also in certain

spiritual activities.

Plants have been used for medicinal purposes long before prehistoric period.

Ancient Unani manuscripts Egyptian papyrus and Chinese writings described the use of

herbs. Evidence exist that Unani Hakims, Indian Vaids and European and Mediterranean

cultures were using herbs for over 4000 years as medicine. Indigenous cultures such as

Rome, Egypt, Iran, Africa and America used herbs in their healing rituals, while other

developed traditional medical systems such as Unani, Ayurveda and Chinese Medicine in

which herbal therapies were used systematically. Traditional systems of medicine

continue to be widely practised on many accounts. Population rise, inadequate supply of

drugs, prohibitive cost of treatments, side effects of several synthetic drugs and

development of resistance to currently used drugs for infectious diseases have led to
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increased emphasis on the use of plant materials as a source of medicines for a wide

variety of human ailments.

Among ancient civilisations, India has been known to be rich repository of

medicinal plants. The forest in India is the principal repository of large number of

medicinal and aromatic plants, which are largely collected as raw materials for

manufacture of drugs and perfumery products. About 8,000 herbal remedies have been

codified in AYUSH systems in INDIA. Ayurveda, Unani, Siddha and Folk (tribal)

medicines are the major systems of indigenous medicines. Among these systems,

Ayurveda and Unani Medicine are most developed and widely practised in India.

Recently, WHO (World Health Organization) estimated that 80 percent of people

worldwide rely on herbal medicines for some aspect of their primary health care needs.

According to WHO, around 21,000 plant species have the potential for being used as

medicinal plants. As per data available over three-quarters of the world population relies

mainly on plants and plant extracts for their health care needs. More than 30% of the

entire plant species, at one time or other were used for medicinal purposes. It has been

estimated, that in developed countries such as United States, plant drugs constitute as

much as 25% of the total drugs, while in fast developing countries such as India and

China, the contribution is as much as 80%. Thus, the economic importance of medicinal

plants is much more to countries such as India than to rest of the world. These countries

provide two third of the plants used in modern system of medicine and the health care

system of rural population depend on indigenous systems of medicine. Treatment with

medicinal plants is considered very safe as there is no or minimal side effects. These

remedies are in sync with nature, which is the biggest advantage. The golden fact
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is that, use of herbal treatments is independent of any age groups and the sexes.

(www.nhp.gov.in/introduction-and-importance-of-medicinal-plants).

One such medicinal fruit is the star gooseberry. It is an ancient fruit, originating in

the tropical climates of Madagascar. Filipino botanist Eduardo Quisumbing explains that

although the fruit came to the Philippines in pre-historic times, the star gooseberries did

not achieve the same popularity there as it did when it spread to Indonesia, Malaysia,

Vietnam and Laos. Nonetheless, countries continue labeling the fruit as their own, hence

its other names such as Sri Lankan gooseberry, Malay gooseberry, and Madagascar

gooseberry. This pungent, sour fruit bears no relation with the more agreeable, reddish

European gooseberry (Ribes uva-crispa).

Today, the fruit grows throughout Asia, parts of Central America, the Caribbean

and parts of South America. Garden hobbyists in Hawaii and Florida dabble with star

gooseberries as well. Pinpointing the fruits arrival in India is difficult because of its

ancient history. Star gooseberrys close relative the amla is distinctly native to parts of

India. How and when these two fruits crossed paths is a mystery. The flesh is juicy,

watery, crisp and densely compact, not unlike the texture of amla.

Intestinal worms may cause severe, life threatening infection, predominantly in

children in tropical and subtropical areas where people often defecate on the ground: in

rural parts of Africa, Middle East, South Asia, Indonesia and Central and South America.

In people from Mediterranean parts of Europe or south parts of North America, they

usually cause a relatively mild disease. Presence of parasites of any kind (including
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intestinal worms) in or on the body is called infestation. Helminth is a medical term for

intestinal worm. (www.healthhype.com).

Herbs that cleanse the body of intestinal worms and parasites are called

anthelmintic (expel) or anti-parasite. They may also be called vermifuges (stunning) or

vermicides (killing). Vermicide is an agent that kills intestinal parasitic worms. (Medical-

dictionary.thefreedictionary.com). The gastro-intestinal helminths become resistant to

currently available anthelmintic drugs therefore there is a foremost problem in

treatment of helminths diseases. Hence there is an increasing demand towards natural

anthelmintic or vermicides. (Investigation of Invitro anthelmintic activity of Thespisia

lampas, Sathish B. Kosalge and Ravindra A. Fursule, 2009).

Raw star gooseberries have pungent, overwhelmingly sour and astringent taste.

Based on this fact, this study was conducted to evaluate the vermicidal property of star

gooseberry (Phyllantus acidus) as other sour tasting fruits like lemon, citron, etc., are

proved to have vermicidal property. ( http://theindianvegan.blogspot.com/2013/03/all-

about-star-gooseberry-phyllanthus.html). The Earthworms belonging to phylum annelida

was used in this study because they are anatomically similar and advanced to the phylum

the Phylum Platyhelminthes and Phylum Aschelminthes that have parasitic worms. Star

gooseberry is an easily available fruit and is also cheap. This study, proven positive is a

boon to all classes of people having helminths diseases who cannot afford to buy the

expensive medicines used to treat the disease. It could also benefit people having

helminths which are resistant to conventional anthelminthics.


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Statement of the Problem

This study aimed to evaluate the vermicidal property of Star Gooseberry

(Phyllanthus acidus) fruit extract.

Specifically, this study sought to answer for the following questions:

1. What is the vermicidal property of Star Gooseberry (Phyllanthus acidus) fruit extract

on the test organism Eudrilus eugeniae in terms of the paralyzing time using the

following treatments?

T0 Positive control (Mebendazole antiox)

T1 100 % of crude extract of Star Gooseberry fruit

T2 75 % of crude extract of Star Gooseberry fruit + 25% of distilled water

T3 50 % of crude extract of Star Gooseberry fruit + 50% of distilled water

T4 25 % of crude extract of Star Gooseberry fruit + 75 % distilled water

2. What is the vermicidal property of Star Gooseberry (Phyllanthus acidus) fruit extract

on the test organism Eudrilus eugeniae in terms of the death time using the above

mentioned treatments?

3. Is there a significant difference in the paralyzing time between and among the groups

subjected to different concentrations of crude extract of Star Gooseberry (Phyllanthus

acidus) fruit?
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4. Is there a significant difference in the death time between and among the groups

subjected to different concentrations of crude extract of Star Gooseberry (Phyllanthus

acidus) fruit?

Scope and Delimitation

This study mainly focused on the evaluation of the vermicidal property of Star

Gooseberry (Phyllanthus acidus) fruit extracts in terms of the paralyzing time and death

time against the organism, namely; Eudrilus eugeniae.

The plant samples were procured from the Vigan public Market. The test

organism used was Eudrilus eugeniae which was obtained from accredited animal

distributors. The control for the experiment used was Mebendazole antiox which was

obtained from the local drug store.

Only the fresh berries of Star Gooseberry plant were used in the experimental

investigation. Plant berries extraction and the Vermicidal property testing was conducted

at the Biotechnology Laboratory in College of Arts and Sciences building, University of

Northern Philippines on 6th March 2017.

Review of Literature and Studies

The following reviews of literature and studies aided the researchers in the

conceptualization of this study.

Medicinal plants

Those plants that have healing properties are termed as medicinal plants or herbs.

Medicinal plants can be simply classified as trees, shrubs, woody perennials, annuals and
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biennials, and climbers. Medical herbalism is the practice of healing with medicinal

plants. The ancient scholars only believed that herbs are only solutions to cure a number

of health related problems and diseases. They conducted thorough study about the same,

experimented to arrive at accurate conclusions about the efficacy of different herbs that

have medicinal value. Most of the drugs, thus formulated, are free of side effects or

reactions. This is the reason why herbal treatment is growing in popularity across the

globe. These herbs that have medicinal quality provide rational means for the treatment

of many internal diseases, which are otherwise considered difficult to cure. Medicinal

plants such as Aloe, Tulsi, Neem, Turmeric and Ginger cure several common ailments.

Lots of consumers are using Basil (Tulsi) for making medicines, black tea, in pooja and

other activities in their day to day life. In several parts of the world many herbs are used

to honour their kings showing it as a symbol of luck. Now, after finding the role of herbs

in medicine, lots of consumers started the plantation of tulsi and other medicinal plants in

their home gardens. Medicinal plants are considered as a rich resources of ingredients

which can be used in drug development either pharmacopoeial, non- pharmacopoeial or

synthetic drugs. A part from that, these plants play a critical role in the development of

human cultures around the whole world. Moreover, some plants are considered as

important source of nutrition and as a result of that they are recommended for their

therapeutic values. Some of these plants include ginger, green tea, walnuts, aloe, pepper

and turmeric etc. Some plants and their derivatives are considered as important source for

active ingredients which are used in aspirin and toothpaste etc. Apart from the medicinal

uses, herbs are also used in natural dye, pest control, food, perfume, tea and so on. In

many countries different kinds of medicinal plants/ herbs are used to keep ants, flies,
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mice and flee away from homes and offices. Nowadays medicinal herbs are important

sources for pharmaceutical manufacturing. Recipes for the treatment of common ailments

such as diarrhoea, constipation, hypertension, low sperm count, dysentery and weak

penile erection, piles, coated tongue, menstrual disorders, bronchial asthma, leucorrhoea

and fevers are given by the traditional medicine practitioners very effectively. Over the

past two decades, there has been a tremendous increase in the use of herbal medicine;

however, there is still a significant lack of research data in this field. Therefore, since

1999, WHO has published three volumes of the WHO monographs on selected medicinal

plants. (www.nhp.gov.in/introduction-and-importance-of-medicinal-plants).

Modern western treatment is different from medical herbalism, but at some point

these two merge. For example, the use of friar's balsam or benzoin tincture for treating

colds, the use of Aloe vera gel for treating sunburn and bruises and the use of cascara or

Senn to relieve constipation. The tendency in modern medicine is to use synthetic drugs,

that eventually were modelled on compounds obtained mainly from plants. Therefore,

whether the plants are used as a whole, or extracts or their synthetics, their discovery

originated from the long term practice of medical herbalism by man.

(kafura12.blogspot.com/p/medicinal-plants).

Helminthic diseases and treatment

Helminths infections, repeatedly entitled helminthiasis are among the most

pervasive infection and a foremost degenerative disease distressing a large proportion of

worlds population. In developing countries, they pose a large threat to public health and

contribute to the prevalence of malnutrition, anemia, eosinophilia and pneumonia. The


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helminths parasites mainly subsist in human body in intestinal tract, but they are also

found in tissue, as their larvae migrate towards them. Most diseases caused by helminths

are of a chronic, debilitating nature; they probably cause more morbidity and greater

economic and social deprivation among humans and animals than any single group of

parasites. Chemical control of helminths coupled with improved management has been

the important worm control strategy throughout the world.

Hookworm disease is a common worm infestation in the developing world caused

by Ancylostoma duodenale or Necator americanus. The illness leads to anemia and

malnutrition. Dracunculiasis is caused by the guinea worm or Dracunculus medinensis,

which is transmitted through contaminated water. It lies burrowed within the skin and

causes severe inflammatory reactions. Loiasis or African eye worm disease is caused by

the filaria Loa loaworm, which is contracted through Deer fly or Mango fly bites. The

adult worms move through subcutaneous tissue towards the sub conjunctiva of the eye.

The illness causes red, itchy swellings in the skin referred to as Calabar swellings.

Cysticercosis is caused by the pork tapeworm or Taenia solium. Symptoms often do not

present for years, but eventually painless bumps develop in the skin and muscles or cause

neurological problems. Echinococcosis is caused by Echinococcus tapeworms. The liver

is usually affected first, followed by the lungs and brain. Liver disease may cause

abdominal pain and jaundice while lung disease leads to breathlessness and coughing.

Filariasis is caused by several species of small nematodes that infect humans through the

bite of an insect. Most including Wucheria spp. and Brugia spp. are Lymphatic filaria

that cause disease by inhabiting and disrupting the function of lymphatics. They are the

most common cause of elephantiasis gross swelling and thickening of skin secondary
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to chronic obstruction of lymphatics. Onchocerca volvulus has a predilection for the eye

and is the cause of river blindness. Schistosomiasis is an extremely important disease in

the developing world with more than 200 million people affected. It is caused by several

species of blood flukes the most important of which are S. hematobium and S.

mansoni. Ascariasis is caused by Ascaris lumbricoides (Vector: Cockroach and Flies)

Symptoms are weakness, Anemia, diarrhea, vomiting, etc.(ezinearticles.com)

Some commonly used anthelminthic include: albendazole (Albenza),

mebendazole (Vermox), niclosamide (Niclocide), oxamniquine (Vansil), praziquantel

(Biltricide), pyrantel (Antiminth), pyantel pamoate (Antiminth) and thiabendazole

(Mintezol). Bitional and ivermectin, used to treat onchocerciasis infestations. Other

antihelminthic drugs, such as diethylcarbamazepine citrate (Hetrezan), used for treatment

of roundworms and other parasites.(medical-dictionary.thefreedictionary.com)

However, development of resistance in helminths against conventional

anthelmintic is a foremost problem in treatment of helminths diseases. Henceforth it is

important to look for alternative strategies against gastrointestinal nematodes, which have

led to the proposal of screening medicinal plants for their anthelmintic activity. (Invitro

anthelmintic activity of Luffa cylindrical leaves in Indian adult Earthworm, Sangh Partap

et al., 2012)

There are some effective natural remedies for treating Ascaris lumbricoides

infection. They can be used as an adjuvant therapy. Coconut oil is an effective home

remedy to flush out roundworms. It has anti-parasitic properties. Drink one to two

tablespoon of coconut oil daily in the morning for few days. Oil contains medium chain
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triglyceride which will help to flush out the parasite from intestine. Unripe papaya

contains an enzyme called papain. It has valuable anti parasitic properties which help to

kill the worm and at the same time facilitate its expulsion. Wormwood is a powerful

anthelminthic natural medicine that has been in use since ancient times. Mix half

teaspoon of wormwood oil and one teaspoon of olive oil to get rid of roundworm. Bitter

gourd, pumpkin seeds, carrots are other natural home remedies that will help in

eradicating roundworms from the intestine.(www.tandurust.com Infections)

Paico and Peppermint both possess vermicidal properties, with the former the

stronger contributor to this medicinal property. (www.herbhealers.com/store/ao-paico-

blend.html). Other plants that have been proved to have anthelmintic property are spongy

guard, lemongrass, lead tree, banana plant, combination of ginger and turmeric, Mimosa

pudica, horse gram and coral tree. (www.motherherbs.com)

Star Gooseberries

Though star gooseberries do not receive as much attention as Amla- the star of

the Phyllanthus genusthese fruits may still be considered super fruits for their

incredible health benefits. Traditionally, star gooseberries are used in India to treat a

number of illnesses. According to the book, Biodiversity in India, these fruits are used

as a blood purifier and appetite stimulant. They are also used to remedy bronchitis,

biliousness, and treat digestive disorders such as urinary concretions, diarrhoea, and piles.

As is the case with Amla, star gooseberry concoctions also act as a liver tonic and blood

enrichment remedy. Another concoction includes making a leaf poultice with added

pepper to treat sciatica and rheumatism.


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Scientific studies also prove the efficacy of star gooseberry as a health remedy. A

2006 study published in Phytomedicine confirms its antibacterial activity when tested for

inhibition against E. coli and staph. A 2007 study published in Molecular Pharmacology

indicates that star gooseberry plant extracts may provide treatment against cystic fibrosis

of the lungs. A 2008 study published in Nature and Science indicates that the leaves have

antimicrobial activities that inhibited growth of E. coli. According to a 2011 study

published in the Journal of Chinese Integrative Medicine, the antioxidants in star

gooseberry fruit have a hepatoprotective effect on the liver. A 2010 article published in

the European Journal of Pharmacology found that leaf extracts reduced blood pressure,

thereby suggesting potent hypotensive properties. A 2012 study published in the Asian

Pacific Journal of Tropical Biomedicine indicates that leaf extracts exhibit strong anti-

inflammatory, analgesic and antioxidant properties. The study suggests that star

gooseberry leaves may remedy oxidative stress, pain and inflammation.

(http://inflibnet.ac.in/ojs/index.php/AUJSAT/article/viewFile/519/504)

In Malaya, the ripe or unripe fruits of Phyllantus acidus is cooked and served as a

relish or made into a thick syrup or sweet preserve. It is also combined with other

fruits in making chutney and jam because, it helps these products to "set".

Often, the fruits are candied, or pickled in salt. In the Philippines, they are used

to make vinegar. In Indonesia, the-tart flesh is added to many dishes as a flavouring

agent. The juice is used in cold drinks in the Philippines (Morton et aI.,1987).

(http://inflibnet.ac.in/ojs/index.php/AUJSAT/article/viewFile/519/504)
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Conceptual Framework

The experimental paradigm that was used in the conduction of this study is

presented in Figure 1.

Input Process Output

By submersion process
and noting the
Star Gooseberry Vermicidal
paralyzing and death
(Phyllanthus Property
time.
acidus) fruit
extract

Figure 1

Figure 1.

The Research Paradigm

As shown in the paradigm, the Star Gooseberry (Phyllanthus acidus) fruit extract

was used in the vermicidal testing. The paralyzing time and death time of the test

organism namely Eudrilus eugeniae was used with justification of the vermicidal activity

of the said plant.

Definition of Terms

For better and clear understanding of the study, the following terms are defined.

Vermicidal Activity: Vermicidal Activity of drugs kill infesting helminths and other

worms. The gastrointestinal tract is the abode of many helminths, although some also live
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in tissues, or their larvae migrate into tissues. This mainly aims in killing helminths and

related worms (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3224400/). In this, it

refers to the vermicidal activity of star gooseberry fruit extract.

Extract: A substance got from something by using a machine or chemicals. (Webster

Third New International Dictionary,2000). In this, it refers to the crude extract obtained

from freshly, handpicked Star Gooseberry (Phyllanthus acidus) using a porcelain mortar

Paralyzing time: To make unable to move or act in a particular time. To impair the

progress or functioning of make inoperative or powerless strict regulations that paralyze

normal activity (http:/www.thefreedictionary.com/paralyzed). In this, it refers to the time

until when no movement of any sort could be observed except the worms were shaken

vigorously or dipped in warm water at 50C.

Death time: Conceptions of death will influence the organism oriented toward seeking

an end to life. When it becomes painful or burdensome in a particular time. The organism

loses its life (http://www.uky.edu/Classes/PHI/350/meaning.htm). In this, it refers to the

until when the worms neither moved when shaken vigorously nor when dipped in warm

water at 50C.

HYPOTHESIS

This study posited the following null hypothesis and tested at 0.01 level of

significance.

There is no significant difference in the vermicidal activity in terms of the

paralyzing time and death time between and among the groups of worms subjected to the

before mentioned treatments.


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Research Methodology

This section presents the design of the study, the materials and the experimental

procedures which were used and strictly followed in the conduct of the study.

Research Design

This study employed the experimental method of the research to gather all the

data needed.

There are three phases:

Phase I: Preparation of the plant extract

Phase II: This phase involved the testing of test organisms with the extracts

prepared.

Phase III: This phase involved recording of the paralyzing and the death time of

the test organisms.

Data Gathering Instruments

The laboratory equipments and materials that were used are the following:

a. Pipette g. Eudrilus eugeniae


b. Petri dishes h. Star Gooseberry fruits
c. Beakers i. Mebendazole antiox
d. Conical flask j. Distilled water
e. Inoculating loop k. Ethyl alcohol
f. Autoclave

Procedure

This portion includes the experimental procedures which were followed during the

conduct of the study. This procedure was adopted and modified from Ajayieoba E.O et al
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as stated from the research entitled Evaluation of Anthelmintic Activity of Leaves of

Paederia foetida of the author Manas Kumar Pal.

A. Procurement of the test organism (Worms)

Earthworm Eudrilus eugenia (Annelida) was collected from the authorized animal

distributors. The average size of Eudrilus eugeniae was 6-8cm. They were washed

with water to remove dirt.

B. Sterilization Process

Autoclaving was used in the sterilization of the apparatus that was used in the

conduct of the experiment.

C. Preparation of the plant extract

The berries of the plant were washed with Ethyl alcohol and the seed were also

removed. The fleshy part of the berries was placed in the Porcelain Mortar to get the

crude extract. The extract thus prepared was filtered using the filter paper.

D. Acclimatization of the test animal

Fecundity, growth, maturation and biomass production will all be significantly

greater at 25C than 15C, 20C or 30C (25C = 77F) of the earthworm (Eudrilus

eugeniae). The habitat for the earthworms was soil at room temperature. The

earthworms fed on decaying roots and leaves. Animal manure is also a good food for

the earthworm.

E. Experiment Property

The experiments were done on adult earthworm Eudrilus eugeniae (Mueller,

1774). 10 ml formulations containing four different concentrations of crude extract


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(25% of the pure extract + 75% of distilled water, 50% of the pure extract + 50% of

distilled water, 75% of the pure extract + 25% of distilled water and 100% of the pure

extract) were prepared. 10ml of the positive control Mebendazole antiox and the

crude extracts were taken in different Petri dishes and four earthworms (same type)

were placed in the solutions respectively. All the test solution and standard drug

solution were prepared freshly before starting the experiments. For each treatment,

three sets of replications were done.

F. Data Gathering Procedure

Time for paralysis was noted when no movement of any sort could be observed

except the worms were shaken vigorously. The worms moved when they were dipped

in warm water at 50C. Time for death of worms was recorded after ascertaining that

the worms neither moved when shaken vigorously nor when dipped in warm water at

50C. Mebendazole antiox (10 mg/ml) was used as reference standard while distilled

water as the control.

G. Statistical Treatment

The following statistical tools were used in the analysis of the data gathered.

Mean. The mean was used to get the average of the paralyzing time and death

time.

Analysis of Variance. The ANOVA was used to determine significant differences

in terms of the paralyzing time and death time, subjected to different treatments.

Scheffe Test. It was used to find out which pairs of the treatments significantly

differed.
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