Antihemintic and Hematological Changes of Natural Plant Carica papaya Seed Extract against Gastrointestinal Nematode

Abstract

The purpose of this experiment was to study the anthelmintic activity of Carica papaya seeds against
Hymenolepis nana in infected mice. In this experiment we use three groups (A, B, and C) of mice, group (A and
B) were infected with eggs of Hymenolepis nana, PCV, Hb and W.B.C counts respectively. Group A mice were
treated with Carica papaya seed extract (0.1ml ⁄mowe) blood and fecal samples were collected on day 0, 7 and
14 after administration of treatment to examine change in blood packed cell volume PCV Hb, W.B.C, fecal egg
counts (FEC). Group C served as a no infected group (negative control). FEC of group A was lower than the
control group, PCV, Hb and W.B.C increased after 7days then return to normal value after 14days. The
histopathological study showed papaya repair the tissue of intestine and return the villa to normal value.
The results recommended using the C.papaya as an aid to control gastrointestinal nematodes infection.

Full text

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Antihemintic and Hematological Changes of Natural Plant
Carica papaya Seed Extract against Gastrointestinal Nematode
Hymenolepis nana
Sabaa T. Mohammed¹*, Nidaa M. Sulaiman¹
1.
Department of Biology, College of Science, AL-Mustansiriya University, Baghdad, Iraq
*E-mail corresponding author: Shebajanabi@yahoo.com
Abstract
The purpose of this experiment was to study the anthelmintic activity of Carica papaya seeds against
Hymenolepis nana in infected mice. In this experiment we use three groups (A, B, and C) of mice, group (A and
B) were infected with eggs of Hymenolepis nana, PCV, Hb and W.B.C counts respectively. Group A mice were
treated with Carica papaya seed extract (0.1ml ⁄mowe) blood and fecal samples were collected on day 0, 7 and
14 after administration of treatment to examine change in blood packed cell volume PCV Hb, W.B.C, fecal egg
counts (FEC). Group C served as a no infected group (negative control). FEC of group A was lower than the
control group, PCV, Hb and W.B.C increased after 7days then return to normal value after 14days. The
histopathological study showed papaya repair the tissue of intestine and return the villa to normal value.
The results recommended using the C.papaya as an aid to control gastrointestinal nematodes infection.
Keywords: Anthelmintic activity Carica papaya, Hymenolepis nana, mice, gastrointestinal nematode.
1. INTRODUCTION
Hymenolepiasis is a disease caused by Hymenolepis diminuta (rat tapeworm) and Hymenolepis nana (dwarf
tapeworm) (Waugh, et al., 2006). Hymenolepiasis has a high prevalence in populations in tropical and
subtropical climates characterized by poor hygiene and poverty
(Melhorn, , et al., 2008; Roberts,
2008)..Hymenolepis nana, is a cosmopolitan intestinal cestode helminthes of the warmer climates, whose entire
life-cycle is completed in the bowel, so infection can persist for years if left untreated (Hardy, et al., 1941).
Hymenolepis nana is the most common cause of all cestode infections and is found globally. In temperate zones,
its incidence is high in children and institutionalized groups. H. nana infections are typically asymptomatic, but
heavy infections by H. nana can cause headaches, weakness, anorexia, abdominal pain, and diarrhea (Chitchang,
et al., 1985). H. nana is the only cestode capable of completing its cycle without an intermediate host. Infection
is most commonly acquired from eggs in the feces of another infected individual, which are transferred by
contamination in food (Smyth, et al., 1989). .
Since ancient times, plants and herbal preparations have been used as medicine. Traditional system of medicine
and folklore claiming that medicinal plants as a whole or their parts are being used in all types of diseases
successfully including antibacterial and anthelmintic, anti-inflammatory etc(Ajaiyeoba, et al., 2001). Carica
papaya, belongs to the family of Caricaceae, and several species of Caricaceae have been used as remedy against
a variety of diseases (Munoz, , et al., 2000; Mello et al.,, 2008). Originally derived from the southern part of
Mexico, C. papaya is a perennial plant, and it is presently distributed over the whole tropical area. Many
scientific investigations have been conducted to evaluate the biological activities of various parts of C. papaya.
(Seigler, et al., 2002). Anthelmintic properties have been reported for latex and seeds from C. papaya when
used in humans and mice(Okeniyi, et al., 2007). The anthelmintic efficacy of Papaya might be due to presence of
proteolytic enzyme such as papain, chymopapain and lysozymes in the latex as well as in leaves (Dakpogan, et
al., 2005).
In the present study, an attempt has been made to screen and evaluate the anthelmintic activity of C. papaya
against H.nana.
2. MATERIALS AND METHODS
Preparation of the C.papaya seeds extract
The seeds were collected freshly from the market of Maskat / Oman, washed with clean water to remove dirt.
The seeds were sundried and later grinded into powdery forms. The C.papaya seed powders were weighed (75g),
and blended into liquefaction in 150ml of distilled water, the mixture was then centrifuged at 1500rpm. The
supernatant was filtered through sterile filter papers into conical flask as the study extract. One milliliter of the
filtrate is expected to contain 0.5g (500mg /ml).
2.1Experimental animals
46 adult healthy male Swiss albino mice, about 4-5 weeks old, weighing 20-25g were maintained under GLP
conditions. 36mice were orally infected with about 100 viable eggs of H.nana per mouse, these infected mice
were maintained until the 6
th
day of infection to complete the life cycle of the parasite.
On the 16
th
day, the mice were divided into 2groups (A, B) with 18mice per group. Group A received no
medication (normal saline). Group B were administrated orally with the aqueous crude extract of C.papaya at

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ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol.4, No.3, 2014
9
(0.1ml /day). These groups were compared with third group (C) contain 9 noninfected mice which used as
control negative.
For all studies, feces and blood were collected on days 0, 7 and 14 after administration of treatment. Blood
was collected by the decapitation method(Dieterich, et al., 1972). anti-coagulated with 1.5mg/ml blood of the
dipotassium salt of Ethylenediaminetetraacetic acid (EDTA), the total and differential leukocyte counts analyzed
using an automatic blood analyzed(Lewis, et al., 1991)., and packed cell volume (PCV) was determined by the
conventional method. Fecal egg count were performed using a modified McMaster technique (Whitlock, et al.,
1948). , also on 14
th
day the mice were sacrificed and the intestine removed, put in (10%) formalin for
histopathological study.
2.2Statistical analysis
Results are expressed as the mean of parameters ± standard error of the mean (SEM), differences between
mean were evaluated using the Anova tests.
3.RESULT AND DISCUSSION
Haematological results showed that the PCV mean values for animals in group A (control group negative) was
(52- 51.5%)after 0.7 and 14days respectively, while in control group positive group C the PCV decreased
between 0 and 14days (45-42%). After treatment with C.papaya seed extract this parameter showed increased
slightly (45-52%) between 0 and14days.
In the case of Hb (g /dl) the mean values for in group A 17(g /dl) and decrease in group C between 0 and 14days
(14.6-13.6) and Hb return increase during this time in group B (14.6-17).
W.B.C. values increase in group C while after treated with C.papaya in group B return to normal value 5.75×10³.
The lymphocyte value exhibited increased in group B, C (7×10³) then decrease during this time period in group
B (5.75×10³). Anova tests showed that when PCV, Hb and lymphocyte W.B.C. compared between groups (B, C)
and control group (A), the differences were significant at p< 0.001(Table 1).
Table1: Haemogram of experimental group during Carica papaya trails
14 days 7 days 1 day Group Parameter
51.5 ±1.29
52 ±0.1
42 ±0.5
52 ± 0.81
51 ± 0.81
43 ± 0.81
52 ± 0.81
45 ± 0.95
45 ± 0.95
A
B
C
P.C.V. (%)
16.8
17 ± 0.01
13.6 ± 0.5
17 ± 0.27
17 ± 0.81
14 ± 0.81
17 ± 0.27
14.6 ± 0.95
14.6 ± 0.95
A
B
C
HB(g/d)
5×10³ ± 0.81
5.75×10³ ±
0.95
7×10³ ± 0.81
5×10³ ± 0.81
5.87×10³ ± 0.25
7×10³ ± 0.81
5×10³ ± 0.81
7×10³ ± 0.81
7×10³ ± 0.81
A
B
C
W.B.C
61.66 ± 2.8
62 ± 1.5
69 ± 0
61.66 ± 2.8
63 ±1
69 ±0
61.66 ± 2.88
68.66 ± 1.15
68.66 ± 1.15
A
B
C
Lymphocyte

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Fecal egg counts showed that the mice in control group heavily shedding eggs in the feces till 14
th
days ,while in
group B after administration of aqueous extracts produced a significant reduction in the fecal egg count after
4days, and the eggs disappear completely after 9days Fig(1).
Fig 1:Egg per gram (EPG)level of infestation before and after treatment with papain
3.1Histological study
The histological study showed that C.papaya plant was able to re-structure of intestinal tissue (Fig 3) has been
able to bring along the length of the villa to normal when compared with control mice (Fig 1), after it suffered
from the palace because of parasite and also led to a lack of preparation of goblet cells (Fig 2).
The aqueous extracts administered in this study caused significant reduction in the worm burden of the mice, the
reduction of helminthes egg loads observed with aqueous extracts of C.papaya maybe attributed to the presence
of papain in capable of digesting bacteria and parasitic cells, hence its use an anthelmintic and
antibiotics(Fajimi, et al., 2005).Anthelmintic is a substance that expels or destroys gastrointestinal worms, all
anthelmintic essentially kill worms by either starving them to death or paralyzing them, because worms have no
means of storing energy. The must sat almost continuously to meet their metabolic needs. Parasite will also die if
they become paralyzed and temporarily lose their ability to maintain their position in the gut. Adu et al.
(2009).reported that C. papaya has certain chemical components that are of high anthelminthic attributes in
poultry with satisfactory efficacy at the dosage of 1200mg ⁄ bird.
Figure 1. Cross- section of a healthy mice intestine high, narrow intestinal villi with a small number of goblet
cells and oligocellular mesenchymal stroma.x400.

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Figure 2. Cross- section of infected mouse's intestine shows shortening of intestinal villi with atrophy of the
villi and depletion of goblet cells.x400.
Figure 3. Section of treated mouse's intestine showing normal like appearance of height of villi and
presence of goblet cells few in number.x250
The mechanism of action of the efficacious plant cysteine proteinases (papaya) is similar, and probably identical
involving digesting and removal of the cuticle. Our results support previous studies (Mursof, et al., 1991).
suggesting that papaya may have potential as an anthelmintic against nematode parasites and too define the
mechanisms of its antiparasitic action.
Worm counts of Hymenolepis nana in mice decreased in response to 1-2g ⁄kg papaya seeds for three consecutive
days (Lamtiur, et al., 1945)., also showed the reduction percentage esin (EPG egg per gram) for papaya seeds
treatment Hymenolepis deminuta in rats, were very high which is 96.8% for 0.6g. kg dose level and 96.2% for
1.2g kg. Dose level (Sapaat et al., 2012).
Fetterer et al.(1978) found an increase in the immunity to Hymenolepis nana infection by transfer from spleen
cells, lymphocytes are essential for the repair mechanism, while Burke et al.(2009) reported there was no
indication that administration of papaya seed led to a reduction in gastrointestinal in lambs.
When papaya latex given in water at dose level of 2, 4, 6and 8g ⁄kg for treated mice infected with 100
Heligmosomoides polygurus found after 3days papaya latex an antiparasitic efficacy of 55.5, 60.3, 67.9 and
84.5 respectively(Satrija, et al., 1995).
also Ameen et al. (2012).showed that the powered and aqueous extract of
C.papaya after its administration, produced a significant increase in packed cell volume, red blood cells,
hemoglobin concentration lymphocyte counts and significant decrease in eosinophil counts. Shaziya and
Goyal(2012) found Carica papaya extract treated mice clearly demonstrated a reduction of larvae for
Ancylostoma caninum, also eosinophil levels were markedly reduced in 24hr after treatment.
Keiichi et al.( 2008)suggest that the oral administration of the fermented papaya preparation to mice may have a
therapeutic potential for the prevention of contact hypersensitive immuno-response, also Melissa et al.(2010).
found no morphological alteration in the integrity of the intestine mucosa after 3months administrated for Wister
rats.
The use of plant material for the management of nematode population is apparently effective and
environmentally friendly compared to synthetic nematicides.

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4.CONCLUSION
From the result it is conclude that the delicious Carica papaya fruit can use it as anthelmintic drug, but we need
to conduct research to detect and extract the active substance and use it in treatment.
5.ACKNOWLEDGMENT
The authors are thankful to Dr.Salim Rasheed AL-ubaidy and Dr.Tubaa Tahier Mohammed for all their help to
accomplish this search.
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