INTRODUCTION parasites, reduce inflammation, lower blood pressure, and lower

INTRODUCTION

Background
of the Study

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     Helminth
is a general term that refers to worms. These helminths are multicellular
eukaryotic invertebrates with either tube-like or flattened bodies exhibiting
bilateral symmetry (O’Donoghue, 2010). They are classified into three groups
according to general external shape and host-organ habitat: Trematodes
(Flukes), Cestodes (Tapeworms), and Nematodes (Roundworms) (Castro, 1996).

Helminths
inhabiting in the gastrointestinal tract are known to be the nematodes
(roundworms).It is revealed that over two billion of the world population are
infected with these parasites. Infection in humans can cause morbidity and
sometimes death by compromising nutritional status and can affect cognitive processes
inducing tissue reactions, such as granuloma. These
intestinal parasites can also cause some gastrointestinal tract problems such
as abdominal pain, diarrhea, anorexia, weight loss, malaise, and provocation of
intestinal obstruction or rectal prolapse (WHO, 2016; Hökelek, 2015). In ruminants,
worms deprive their hosts such as pigs of nutrients, thus, affecting the growth
and feed efficiency. Severe infestation can lead to condemnation and loss of
carcass value (Jacela, 2009).

Throughout the world, the important worm
control strategy has been the chemical control of nematodes coupled with
improved management. Yet, the problem in the treatment of parasitic diseases is
the development of resistance in nematodes against conventional anthelmintics (Partap
et al., 2012). Resistance has arisen to all of the major families of broad
spectrum anthelmintics such as the benzimidazoles, levamisole, and the other
nicotinic agonists, in addition to the avermectins and milbemycins (Sangter and
Gill, 1999 cited in Shalaby, 2013). Hence, it is essential to look for
alternative anthelmintic.

Papaya (Carica
papaya) has become common in the society for its various contribution to
the field of medicine. It is a giant herbaceous plant resembling a tree but not
woody in the Caricaceae family that originated in Central America and is now
grown in tropical areas worldwide for its large, sweet, melon-like fruit
(Courteau, 2017). In some developing countries, the traditional use of papaya
is being investigated as an alternative to commercial treatments for a range of
ailments. It has been used to treat diabetes, as birth control, as an
antiseptic, antimicrobial, or diuretic, to control parasites, reduce
inflammation, lower blood pressure, and lower cholesterol. While there are only
limited data to support most of these uses, there are some evidences for
healing bed sores, and other wounds and in treating intestinal worms in humans
(Wolters Kluwer Health, 2009).

Certain parts of this plant have been tested for
their anthelmintic activity. The papaya seeds, be it in powder or aqueous
extract, reduced the
helminth eggs positively in infected commercial layers (Ameen et al., 2011). To further determine the
anthelmintic activity of papaya, a decoction of its mature leaves were tested
in this study against the gastrointestinal parasites of Philippine native pigs.

 

Statement
of the Problem

     Generally, the study aimed to prove the
anthelmintic potential of papaya leaves decoction against gastrointestinal
parasites of Philippine native pigs.

Specifically,
it aimed to answer the following:

1.
What is the pre-treatment and post-treatment
egg count per gram?; and

2.
Is there a difference between the mean total
percentage reductions of the treatments?

 

Significance
of the Study

The results of the study would benefit the pig
raisers as they may be provided with new, cheaper, and effective
anthelmintic organic drugs which can keep their animals clean and healthy and
as organic as possible. The study may also benefit the papaya growers as
they may produce another use of the said plant making the demand possibly
higher.  This may benefit the society
as new alternative medicine will be introduced. Lastly, the result would serve
as a benchmark information for future researchers.

 

Scope
and Delimitations

     The study was delimited in proving the
anthelmintic potential of papaya leaves decoction against gastrointestinal
parasites of Philippine native pigs regardless of their sex. All pigs used were
1-4 months old. Fecal analysis through McMaster Floatation and Egg Counting
Technique was utilized as diagnostic procedure to examine the animal feces.

     The study was conducted at College of
Veterinary Medicine, Pampanga State Agricultural University, Magalang, Pampanga
on September 2017.

 

Definition of Terms

Anthelmintics
refers to agents used to eradicate intestinal worms
(helminthes) from the body (Drugs.com, 2017).

Anthelmintic Resistance refers to the heritable (and therefore genetic) ability of the
worm to survive a dose of anthelmintic which would normally be effective. It could
also be described as ‘drug tolerance’ in worms (SCOPS, 2012).

In vitro refers to
the technique of performing a given procedure in a controlled environment
outside of a living organism (Autoimmunity Research Foundation, 2012).

In vivo refers to
the technique of performing a given procedure inside of a living organism.
(Autoimmunity Research Foundation, 2012).

 

REVIEW OF THE RELATED LITERATURE

Taxonomic
Classification

     Family Name: Caricaceae

          Scientific Name: Carica papaya

Common Name:
Papaya

                                          

Botanical
Description

     Carica papaya or commonly known as Papaya is a huge herb
which was introduced from America and widely distributed throughout the
Philippines. It has one erect unbranched trunk. It is a fast-growing tree that
can reach 3-7 meters in height. The tree is characterized by its large green
leaves that have 7-9 deeply palmate lobes with elongated hollow petioles
confined in spiral at the top of the trunk. The trunk
and leaves both contain copious white milky latex. The flowers are 5-petalled
and are fleshy, waxy, and slightly fragrant. The fruit, in general, has
smooth skin, green when young which turns yellow when ripe. It is indehiscent,
subglobose, obovoid, or oblong cylindric. The fruit is rich in latex when it is
unripe or green-colored. As it ripens, it becomes light-deep yellow externally,
the thick wall of succulent pulp becomes aromatic and yellow or orange in
color, and contains black seeds coated with a gelatinous and transparent aril
(Morton, 1987; Stuart, 2016; Medical Health Guide, 2017).

Nutritional
Content

     Carica papaya contains many biologically active
compounds. Papaya leaves, fruit, stem, and roots all contain
proteolytic enzyme, papaytin, phytokinase, malic acid, and calcium mealeate.
Fresh latex contains chymopapain. Both leaves and fruit yield saccharose,
dextrose, levulose, malic acid, and citrates. Carpaine and carposide are only
found in the leaves. The fruit also contains papain. Volatile oils are found in
the seeds (Stuart, 2016). A
medium-sized papaya fruit contains macronutrients such as protein,
carbohydrates, fats, and dietary fiber, water-soluble vitamins such as vitamin
C and B, fat-soluble vitamins such as Vitamin A, D, E, and K, minerals such as
Calcium, Chloride, Copper, Iron, Magnesium, Manganese, Phosphorus, Potassium,
Selenium, and Sodium, and some amino acids (Mateljan et al., 2017 from The George Mateljan
Foundation). According to
Rashed et al. (2013), extracts of the
polar parts of Papaya through various tests contain triterpenes, carbohydrates,
flavonoids, alkaloids, and tannins. As a whole, its phytochemical composition
include unique protein-digesting proteolytic enzymes like Papain and Chymopapain, crytoxanthin,
Carposide, Glucosynolates, Benzyl isothiocyanate, papaya oil, Monoterpenoids,
Flavonoids, Alkaloids(Aravind et
al., 2013).

 

Health
Benefits

     Papaya is known for its many medicinal uses
and health benefits. It contains zeaxanthinin which aids in age-related macular
degeneration. Papaya also helps in asthma prevention due to its high amount of
certain nutrients such as beta-carotene. Vitamin K in papaya helps in improving
calcium absorption resulting to strengthening and rebuilding of bones.
Different studies also concluded that papaya aids types 1 and 2 diabetes.
Papain, an enzyme in papaya, aids in digestion. The fiber, potassium, and
vitamin content of papaya help prevent cardiovascular diseases. Papaya contains a very important
and versatile nutrient called choline which aids bodies in sleep, muscle
movement, learning, and memory. It also helps in maintaining cellular membrane
structure, aids transmission of nerve impulses, assists in absorption of fats,
and reduce chronic inflammation (Ware, 2017). The rich fiber content of
papaya lowers the risk of cancer and prevents oxidation of cholesterol.
Vitamins A and C help in boosting the body’s immunity (Bora, 2016).

Native Pigs

 

     Philippine native pigs are distinguished by
their well-built body, with a coarse, thick, black color coat with a dispersion
of silvery white color fur on their sides. Firm fur form at the top of their
head running down to the central point of the back which is noticeable
especially during breeding season on male pigs when it forms a famous mane over
their neck and head crest. The tail of an average length is also covered with
thick elongated fur up to its tip which is used to drive away insects and to
indicate their mood (Lookseek, 2017).

Philippine native pigs have the ability to grow
and reproduce under adverse conditions. These animals are budget-friendly
especially to farmers of small scale who cannot aid in buying commercial feeds
because low-cost production inputs are required in terms of housing and
feeding. These native animals can become an additional source of income for
small-scale farmers in rural areas (Brion, 2016).  Despite the low maintenance cost of living of
these pigs, many illnesses may infest the animals as well as parasites.
External parasites such as mange mites, lice, and fleas accompany irritation
and discomfort to the pigs and may cause parakeratosis. On the other hand,
internal parasites such as ascarid, lungworms, whip worms, nodular worms, and
round worms may cause death to the host. Meanwhile, herbal plants can act as
substitutes to ease common diseases and illnesses (Esminger, 1970). Such
infestation may show symptoms through coughing, poor growth, hairiness,
vomiting, blood in feces (rare), diarrhea, and distended belly (Sawyer and Cox,
2015 from Mini Pig Info).

 

Gastrointestinal Parasites

Helminths
are parasites that live and feed on a living host to be nourished and
protected, while causing nutrient absorption, weakness and disease in the host.
These worms and larvae live in the small intestines and referred to as
intestinal parasites (Mandal, 2014). They are grouped according to their shape,
and the host organ they inhabit. They develop through egg, larva, and adult
stages (Castro, 1996). Major groups of helminths include platyhelminths,
acanthocephalins, cestodes, trematodes, and nematodes.

Nematode or mostly known as roundworms are known for having
long thin unsegmented tube-like bodies with anterior mouths and longitudinal
digestive tracts. Their fluid-filled internal body cavity (pseudocoelum) acts
as a hydrostatic skeleton providing rigidity. Worms use longitudinal muscles to
produce a sideways thrashing motion. Adult worms form separate sexes with
well-developed reproductive systems (O’Donoghue, 2010).

Intestinal Parasitism

Parasites
inhabiting the digestive canal and biliary and urinary systems produce eggs,
larvae, or cysts that leave the body of the host by way of feces or urine.
Occasionally even adult parasites may be seen in feces especially when the host
has enteritis. Parasitic worm eggs or larvae from the lower respiratory system
are usually coughed into the pharynx and swallowed and they as well appear in
feces (Sloss and Kemp, 1996 cited by Velasco, 2006).     These
worms deprive their hosts of nutrients such as pigs, thus, affecting the growth
and feed efficiency. Severe infestation can lead to condemnation and loss of
carcass value (Jacela, 2009).

 

Fecal
Analysis

     Intestinal
parasitism can be determined through Fecal Analysis since the parasites live in
the gastrointestianl tract of the host. Fecal analysis is done using a
microscope and can either be in an in-house lab or at an off-site laboratory.
The test requires fresh fecal samples. Fecal analysis can be done in two ways –
floatation method or centrifugation. Both methods require a mixture of a sample
of a certain mass and a floatation solution which is mostly saturated salt. The
floatation solution due to its higher specific gravity allows the eggs to float
at the top. The parasites are then identified under a microscope (Glanbrook
Veterinary Services, 2016).

 

Fecal Egg
Count Reduction Test (FECRT)

 

     The most
recommended method in exploring the anthelmintic potential of a compound is the
fecal egg count reduction test (FECRT). This is easy to conduct, suitable for
ruminants, horses, and pigs as well as for all types of anthelmintic. Due to
these, it has already been standardized which allowed its broad use (Sanchez et al., 2002).

 

Levamisole

Levamisole
is an anthelminthic drug that has been tried experimentally in rheumatic
disorders where it apparently restores the immune response by increasing
macrophage chemotaxis and T-lymphocyte function. Paradoxically, this immune
enhancement appears to be beneficial in rheumatoid arthritis where dermatitis,
leukopenia, and thrombocytopenia, and nausea and vomiting have been reported as
side effects (From Smith and Reynard, Textbook of Pharmacology, 1991). Levamisole, and pyrantel and morantel, are
agonists at nicotinic acetylcholine receptors of nematode muscle and cause
spastic paralysis (Martin, 1997). However, resistance against this drug has
been recorded in 1985. A study by Hembry et al., the levamisole treated group in the study had an increase
of EPG from 3 to 483. In the end of the study, EPG of the levamisole treated
group increased by 627%.

 

RELATED
STUDIES

Anthelmintic
Plants

 

     For the developed and developing countries,
herbal medicines are in great demand as they may serve for primary health care
because of their wide biological and medicinal uses, higher safety margins, and
less expensive. Hence
are traditionally used in various parts of the world to cure different diseases
(Kamboj, 2000). From ancient time different plants are known to possess
significant anthelmintic activity against these invasive types of worms and can
be effectively used in the treatment of worm infections (Tambe and Bhambar,
2014).  

     Chaluvaraju
et al. (2015)
stated that most of the plants which were tested for their anthelmintic
activity belong to families such as Schrophularaceae, Fabaceae, Polygonaceae,
Capparidaceae, Apocyanaceae, Myrtaceace, Caesalpinaceae, Juglandaceae,
Amaranthaceae, Araceae, and Rubiaceae.

Anthelmintic plants offer a traditional
alternative to manufacture anthelmintics that is both sustainable and
environmentally acceptable. Such plants could have a more important role in the
future control of helminth infections in the tropics (Hammond, Bishop, and
Fielding, 1997).

     In most of the cases in in vitro tests, adult earthworms were
used as test organisms in screening the anthelmintic activity of certain plants
due to the anatomical and physiological resemblance to intestinal roundworm
parasite of human beings. On through survey, it has also been concluded that
methanol, ethanol, and aqueous extracts showed significant anthelmintic
activity (Chaluvaraju et al., 2015).

 

Papaya (Carica papaya) as Anthelmintic Plant

Three studies have proven the anthelmintic
activity of different papaya parts. Papaya latex has been used in the study of
Satrija et al. (1995) where five
groups of mice were infected with 100 Heligmosomoides polygyrus. Four (4)
groups were treated at dose levels of 2, 4, 6, & 8 g latex per kilogram
bodyweight, respectively. One group served as negative control (group A). At
day 25 of the experiment, all animals were necropsied to determine the worm
counts. The latex exhibited an anthelmintic efficacy reducing the count by
55.5, 60.3, 67.9 and 84.5% in groups B, C, D and E, respectively. The potential
role of papaya latex as an anthelmitic plant against patent intestinal
nematodes of mammalian hosts is suggested. Thus, papaya latex was used by
Nansen et al. (1994) to determine
whether it can be an alernative anthelmintic to 16 naturally infected pigs.
Four (4) treatment groups were observed but only 3 groups were administered
with papaya latex at dose level of 2, 4, and 8 g latex per kilogram bodyweight
and 1 group served as untreated group. Results exhibited reductions at 39.5,
80.1 and 100% in groups B, C, and D, respectively. It was observed that the
group that was given the highest dose experienced mild diarrhea. Otherwise, no
clinical and pathological changes were observed in treated animals. The last
study by Ameen et al. (2012) assessed
the anthelminthic potential of papaya leaves crude and aqueous extracts in 40
naturally infected Isa Brown commercial layers. The birds were randonly grouped
into 4 with 1 group (group A) being untreated. Blood and fecal samples were
collected 2 weeks after the treatment to evaluate for hematological values and
fecal egg counts, respectively. The two extracts showed a significant increase
in packed cell volume, red blood cells, hemoglobin concentration, lymphocyte
counts and a significant decrease in eosinophil counts. Fecal egg count significantly
drecreased but it was more pronounced with the aqueous extract.

 

Anthelmintic
Phytochemical Components

 

Different preliminary
phytochemical tests prior to anthelmintic tests of several plants suggest that
it may be flavonoids, tannins, saponins, alkaloids, terpenes, steroids, or
glycocides which are responsible for the anthelmintic effect of the plants.

In the studies of Das et al. (2011) and Ranjan (2013), the tannins of Tamarind produce
anthelminitic actitvities as they bind to free proteins in the GI tract of host
animals or glycoprotein on the cuticle of the parasite thereby cause deaths. These compounds are known to be biologically active and therefore aid
the antimicrobial activity. Tannins have been found to form irreversible
complexes with highly rich protein resulting in the inhibition of cell protein
synthesis. Tannins are known to react with protein to provide difficult tanning
effect which is important for the treatment of influenced or ulcerated tissues.
Herbs that have tannins have the main component astringen which are used for
treating intestinal disorder such as diarrhea and dysentery.

A study to support this is conducted by
Prevot et al. (2003) in France which
made use of condensed tannins as anthelmintic to goat that were infested with
adult Haemonchus
contortus. The
administration of tannins was associated with a significant decrease in egg
excretion in the fecal samples of the test animals, which persisted until the
end of experiment. According to McCarry et al. (2001),
a more specific compound is present in papaya that is responsible for its
anthelmintic potential. The papaya extract contain benzyl isothiocyanate that
is predominantly or solely inhibits energy metabolism and affects the motor
activity of the parasites that makes it anthelmintic.

Literature Synthesis

 

     Different plants have been tested for their anthelmintic activities.
Papaya leaves, due to them being easily accessible, have been tested repeatedly
using different test organisms. In the aforementioned studies which involved
the plant, all of the tests showed successful results as they can serve as
alternative anthelmintic. Different literature say that the plant’s tannin
content (Das et al. 2011 and Ranjan,
2013) and the Benzyl isothiocyanate and Papain which Papaya seeds contain (McCarry et al.,
2001) make Papayas an effective anthelmintic. Thus, the researchers were
prompted to extend a research on effective and organic anthelmintic drug on
native pigs.