Effect of Leaves Extract of Carica papaya, Vernonia amigdalina and Azadiratcha indica on the Coccidiosis in Free-range Chickens
An alternative coccidiosis treatment in response to the side effects of routinely use of conventional anticoccidial drugs was investigated on 600 E. tenella experimentally infected day-old Harco chicks in a factorial design at the Benin Agronomic Faculty Research Station. The effect of Carica papaya, Vernonia amygdalina and Azadiratcha indica dry leaf powder in the conventional feed (15% incorporation) was evaluated. C. papaya, the most ingested plant reduced the infected untreated control OPG down to 53% and V. amygdalina, down to 35%, even though it was less ingested than C. papaya. However, this reduction constituted 56 and 37% (C. papaya and V. amygdalina) of the conventional coccidiostatic effectiveness. The OPG reduction might be ascribed on the hand to the bitterness and on the other to the chemical compound, papaine and Vernoside. A. indica was less ingested due to it higher bitterness, repellency and toxicity. Consequently it exalted the disease and produce more oocysts than the control group. The plant OPG reduction mechanism and the accepted leaf powder dose need to be investigated.
Coccidiosis is one of the most expensive and common diseases of poultry production systems in spite of advances in chemotherapy, management, nutrition and genetics (McDougald et al., 1987). It remains a big concern for the commercial chicken production because of the high costs to control the disease. Williams (1999) estimated the total cost of chicken coccidiosis control in UK in 1995 at £38.5million. McDougald (2003) reported that the current expense for preventive medication exceeds $90 million in the United States and more than $300 million worldwide. Coccidiosis may strike any type of poultry in any type of facility (McDougald, 2003). The disease is also present in free-range production systems; in Zimbabwe coccidiosis was reported to be the most important endoparasitic disease and a second direct and indirect cause of loss in Village chicken after Newcastle disease according to Kusina et al. (2004).
The discovery of sulfanilamide by Levine (1939) has led to the use of anticoccidial feed additives more recently, polyether ionophorous antibiotics, have also been developed and used (Matsuda et al., 1989). The routinely use of those drugs in one hand has led to parasite drug resistant strains (Long, 1982) and on the other hand, prejudicial to consumer health because of drug or antibiotic residues in poultry products (Youn and Noh, 2001). In the recent years, the pharmaceutical industry has shown little interest in developing new products for anti-parasitic use and it is unlikely that we will have replacements for the products already available (McDougald, 2003). The search of alternative tools such as medicinal plants could be consequently envisaged in producing ecological meat, particularly in free-range production systems where the so-called organic product is increasingly demanded by consumers.
In Benin the use of medicinal plants in human malaria treatment is a common practice. Various sour and bitter leave or roots are used for medicinal purposes, but the efficacy is not documented (Personal observation). In many tropical and subtropical countries, numerous traditional medicinal plants have been used for centuries (Dharma, 1985; Perri, 1980). According to Lal et al. (1976), preparation of B. frondosa, Carica papaya, Momordica charantia and Sapidus trifoliatus have been found effective in vitro against poultry Ascaridia galli. In Senegal, farmers have traditionally used such plants to treat their chickens against endoparasites, for example, Capsicum sp. extracts and the leaves or barks of Azadirachta indica are added to drinking water and given to birds. In Cameroon, Agbédé et al. (1995) reported good results from the use of plants such as Kalanchoe crenata for coccidiosis and papaw (Carica papaya) leaves for diarrhea, while the use of human medicines (especially antibiotics, Ampicillin, Tifomycin) achieved no success. Huffman and Wrangham (1994) showed a positive effect of Vernonia amygdalina on chimpanzee infected with parasites.
The objective of this study was to try out the effect of Azadiratcha indica, Carica papaya and Vernonia amigdalina on infections with Eimeria tenella in chickens.
MATERIALS AND METHODS
Study Design and Duration
The study was designed as a factorial design with anticoccidials and oocysts
doses as group factors. The duration was 2 months with 3 replications.
Eimeria tenella infective oocysts (3000 and 30000 oocysts doses in 2%
potassium dichromate solution) were provided by Institut de Recherche Agricole
in Tours, France and conserved in a fridge at 0-4°C.
600 day-old Harco male chicks were provided by the hatchery Poussin du roi
and reared in the Department of Biological Sciences, King Abdulaziz University
Faculty of Science, Jeddah. Saudi Arabia. The breeders were imported from France
by the hatchery. On the day of delivery all chicks were vaccinated against Newcastle
disease and Infectious bronchitis.
Chicks were reared in a chick room before transferring them to the treatment
rooms at the age of 21 days. The 200 chicks were placed in 10 different rooms.
Chicks in 5 rooms were orally challenged with 3000 E. tenella oocysts
doses and the other 5, by 30000 E. tenella oocysts.
The experimental feed was a starter feed with non-anticoccidial additives.
The feed was mixed with dry leaves powder at 15% incorporation level. The leaves
were Azadiratcha indica (AI), Carica papaya (CP) or Venonia
amigdalina (VA). Fresh leaves were collected and dried in rooms. The exposure
to the sun was avoided in order to ensure the leaf chemical constituents conservation.
The 10 chicken rooms were allocated to 5 groups. Each group comprised 2
rooms: the first was the 3000 oocyst dose infected chicks and the second 30000
oocysts dose infected chicks. Three groups received dry leaves feed additive,
the 4th one was treated with a coccidiostatic (AnticoxR) in drinking
water throughout the experiment. The treatment lasted for 4 weeks after E.
tenella experimental infection. The last group was the infected untreated
The body weight gain, the cumulative feed intake, the feed conversion ratio,
the survival rate, the number of animals with bloody diarrhea and the lesion
score were investigated during the 4 weeks following the E. tenella experimental
infection. Feces were collected from the chick rooms around the water container
in the morning to reduce the litter content. The oocyst excretion was daily
monitored during the 14 days patent period.
420 feces samples were taken to count the oocysts using the Mc Master technique.
Dead birds were examined and the lesion score obtained using the scoring method
described by Johnson and Reid (1970).
The number of oocysts counted from the McMaster chamber multiply by 200
gave the OPG. This number of oocysts is the mean of the oocysts found in the
cells in the Mc-Master Chamber.
Statistical Analysis and Graphs
The descriptive analysis was made using the Proc Univarite Model (SAS version
8.2). The hypothesis was tested using the Proc Mixed Model (SAS, V. 8.02). The
survival rate comparisons were tested in a Chi-square model using the GraphPad
Prism Program (Version 4.02). The graphs were made using the chart facility
of MS Excel 2000.
The survivabilities of V. amigdalina (64%), C. papaya (72%)
and A. indica (61%) treated chicks were statistically similar; but significantly
lower than the coccidiostatic group survivability (84%) and higher than survivability
of the control group (59%) (Table 1a and b).
The post-mortem of dead birds revealed hemorrhages or thickening whitish mucosa
and cores of clotted blood in the caeca which specific for Eimeria tenella
infection. The number of bloody diarrhea and the lesion score followed almost
the same trend.
|Table 1a :
||21 day-old chicks parasitological results during 4 weeks experimental
E. tenella post infection period (treatment effect)
|Table 1b :
||21 day-old chicks parasitological results during 4 weeks experimental
E. tenella post infection period (oocyst doses effect)
|P: mean proportion, SEM: Standard Error on the Mean, OPG:
Oocysts Per Gram, Columns that is not sharing superscript letters are statistically
different in Chi-square test (survival rate, Graph pad prism), Blood in
feces (genmod procedure, SAS), Lesion score and OPG (proc mixed hypothesis
The coccidiostatic treated chicks had a higher survival rate (83%) and no bloody
diarrhea was observed in their rooms. Significantly lower survival rate (58%),
higher number of bloody diarrhea (4.6) and higher lesion score (3.1) were observed
in the control group compared to the 4 treated groups Lower mean OPG was observed
with coccidiostatic treated chicks (3136) that constituted 6% of the control
OPG and higher OPG in A. indica treated group constituting 127% of the
control OPG. The OPG reduction induced by C. papaya leaves treatment
was 53% of the OPG of the infected untreated control group, which represented
56% of the conventional coccidiostatic treatment efficacy. Similarly the OPG
reduction caused by V. amigdalina leaves treatment was 35% of the OPG
of the control group, which represented 37% of the conventional coccidiostatic
treatment efficacy. No significance OPG difference was observed between the
C. papaya and V. amygdalina treated groups. The peak of OPG (Fig.
1) observed in the 7th day of oocysts excretion in the control group was
delayed about 2 days in the C. papaya group (on day 9) and one day in
the V. amigdalina group (8th day). The effect of oocyst doses was not
significant on the parasitological results (p>0.05).
Chick Growth Performance
V. amigdalina and A. indica infected treated chicks
exhibited significantly growth performance (weight gain and feed efficiency
compared to C. papaya, Coccidiostatic treatment and the control group
(Table 2a and b). Higher weight gain (388
g) and daily body gain (10 g) were observed with coccidiostatic treated chicks
and lower values (weight: 213 g and DBG: 3 g) with A. indica treated
chicks. The cumulative feed intake was higher in C. papaya (914 g), coccidiostatic
treated chicks (905 g) and the control group (896 g) than in V. amigdalina
(511 g) and A. indica treated chicks (494 g).
||OPG evolution during 14 days patent period
||21 day-old chicks growth performance during 4 weeks experimental
E. tenella post-infection period (treatment effect)
|*, ** and *** are significant at the level of p<0.05, p<0.01
and p<0.005, respectively
||21 day-old chicks growth performance during 4 weeks experimental
E. tenella post-infection period (oocyst doses effect)
|PI: Post Infection, DBG: Daily Body Gain, CFIW4: Cumulative
Feed Intake at Week 4, FCR: Feed Conversion Ratio Columns that are not sharing
superscript letters are statistically different in proc mixed hypothesis
The feed conversion ratios of almost all experimental groups were statistically
similar, with exception of the A. indica group, which disposed of a higher
feed conversion ratio (7 g feed/g weight gain). The effect of the oocyst doses
was not significant on chick growth performance (p>0.05).
C. papaya leaves powder in the feed was the most ingested plant because
of less bitterness of the dry leaves and the presence of carotene (vitamin A)
that can enhance the feed palatability and consequently impact positively chick
growth performance (Eramus et al., 1960). However, it did not influence
the chicks growth performance when compared with the coccidiostatic treated
chicks and the control. The papaya treatment reduced significantly the OPG down
to 53%. This result is in line with previous experiments showing similar activity
of papaya latex against infections of Ascaridia galli, in chicken (Mursof and
HE, 1991). Satrija et al. (1994), by trying out the efficacy of papaya
latex against Ascaridia suum in pigs, found a decrease of the eggs produced
(by 99%) and the number of adult worms (by 80%). Purwati and He (1991) indicated
the effectiveness of papaya latex on the worm organism as well as the egg infectivity.
According to these authors, the anthelmintic activity of papaya latex might
be ascribed to proteolytic enzymes such as papain, chymopapain and lysozyme
present in the latex as well as in the leaves, seeds and roots (Winarmo, 1983).
Obviously, worms or parasites in general, being a proteinic organism can be
digested by the papaine. The vitamin A supplementing feed showed a great reduction
of excreted E. tenella oocysts (Eramus et al., 1960; Coles et
al., 1970; Singh and Donovan, 1973). Furthermore, C. papaya is known
to have an anti-inflammatory properties, certainly due to it riches in Vitamin
A, used against tumors, ulcers and can accelerate wound healing (Beuth et
al., 2001). In the current study, the lesion score and the bloody diarrhea
recorded was middle compared to the V. amygdalina and A. indica
treatment and the control. The OPG reduction induced by C. papaya dry
leaves powder treatment might be ascribed in one hand to the direct E. tenella
protozoits digestion in the caeca by a synergistic action of pancreas chymotrypsin
and papaine. The anti-inflammatory property of the C. papaya concentrated
vitamin A on the other hand might act in caecal epithelium cell protection,
detrimental to the coccidia reproductive activities. Nevertheless, the observed
OPG reduction constituted only 56% of the coccidiostatic treatment efficacy
and the Survival rate was significantly lesser than the later.
Venonia amygdalina and Azadiratcha indica
The feed containing V. amigdalina and A. indica dry leaf
powder was less ingested. Consequently, the treated chicks exhibited a very
poor growth performance. This was almost 50% of the growth performance observed
in the C. papaya group. The drying process did not reduce the bitterness
of these 2 plants, as it was the case with C. papaya. A. indica is reputed
for it toxicity and the smoke from the fresh burned leaves is traditionally
used to repel mosquitoes (personal observation). Richard et al. (2003),
by evaluating neem extract for use against the turnip root fly (Delia floralis
fabi) indicated a repellent effect that reduced the fly oviposition. The
toxicity of the neem leaves and V. amygdalina leaves in lesser extent
(Huffman et al., 1996) might repel the chicks from feeding, detrimental
to the growth performance. Even though V. amygdalina was less consumed
it OPG reduction effect revealed to be significant (35%) and constitute 37%
of the conventional coccidiostatic efficacy. Huffman and Wranghan (1994) noted
that multiple parasitic affected chimpanzees chew V. amygdalina pith.
V. amygdalina bioassay examination indicated an antiparasitic effect. Further
analysis done by Ohigashi et al. (1994) revealed chemicals categorized
as Sesquiterpen lactones and steroid glycosides, which are known for their anti-tumor
activity. Specifically vernoside B1 and vernoniol B1 extracted from the pith,
suppressed movement and egg-laying activity in a bioassay of Schistosoma
japonicum, a parasitic worm and have proven effectiveness against drug-resistant
malarial parasites a protozoan organism. The toxicity of V. amygdalina can
be directly prejudicial to the E. tenella sporocysts. The bitterness
of the medicinal feed might enhance the gastrointestinal enzymes (chymotrypsin)
production and the digestion of the sporozoits. In addition the vernoside B1
or vernoniole B1 reduction activities against Eimeria is conceivable. However,
further studies need to be carried out to elucidate theses suggestions. Contrary,
the A. indica powder increased the oocysts production of the control
group up to 127% and aggravated the disease. The necropsy from the neem leave
treated chicks showed enlarged yellowish liver (observation of the author but
not quantified), which might be ascribed to the high toxicity of the plant.
Such condition can also weaken the chick immune system and cause the organism
natural defense reduction.
Carica papaya leaves and venonia amygdalina have showed a positive effect on OPG reduction compared to the neem leaves treatment OPG, where the value passed largely the control one. Their action might be ascribed to the bitterness or toxicity. In addition it might be in keeping with enzyme activities that can digest and eliminate the sporozoits. The protection of caecal epithelium in the case of C. papaya is suggestible. Nevertheless the E. tenella OPG reduction observed was almost 50% of the conventional coccidiostatic efficacy. Either ways, some researches need to be carried out to precise the oocyst reduction mechanism in the ceaca as well as the acceptable incorporation level of A. indica and V. amigdalina dry leaves powder.
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