1. Introduction
Various constraints make delivery of veterinary services and industrially manufactured veterinary drugs either unavailable or unaffordable in many rural areas of Ghana and in other developing countries. The use of indigenous knowledge and medicinal plants for animal health interventions is, therefore, a reality in many of these rural communities. Pawpaw extracts have been used to treat at least 40 human diseases (Lewis and Elvin, 1977; Mezhlumyam et al., 2003). The medicinal value of pawpaw most likely lies in the 142 phytochemical components that Duke (2001) lists as contained in pawpaw. The perception about pawpaw (Carica papaya) seeds, offered to chicken in rural communities in Ghana is often a paradox. Whereas in some communities, pawpaw seeds have often been thrown to domestic chicken to alleviate coccidiosis, in some other areas, pawpaw seeds have been wrongly suspected of causing fowl pox because of the semblance of the seeds to pox lesions. Harsha and Chinoy (1996) and Satrija et al. (1995) cite pawpaw as a strong amoebicide. That pawpaw has been suggested as an amoebicide circumstantially suggests that extracts of pawpaw must be able to affect coccidia (Eimeria spp.), the protozoon that causes coccidiosis in chicken, resulting in economic losses in poultry rearing through morbidity, mortality and treatment costs (Braunius, 1987; Reid, 1990, Ruff and Allen, 1990; Hofstad, 1984; McDouglad, 1993). Since chicken is a dependable quick protein source and even a store of economic value in rural communities in rural areas of Ghana and other developing countries, a rural dweller-friendly intervention taking advantage of the indigenous knowledge on the use of pawpaw in the treatment of coccidiosis will be of a great help as a first aid for coccidiosis. It is in view of this that this study was conducted to validate and assess the potential of the use of aqueous extracts of seeds, roots and leaves of pawpaw as anticoccidia agents in chicken and with a method that the ordinary rural farmer can easily use in a developing country.
2. Materials and methods
1. Experimental birds
The use of chicken for this study was in accordance with internationally accepted principles for animal use and care. From a flock of one thousand 5-week old Lohman Brown chicken, 84 were randomly selected and used in this study. The birds had undergone routine prophylactic immunizations for their age (Marek, New Castle, Gumboro and Fowl Pox), fed on commercial grower’s feed and drank water ad libitum during the study. The experimental birds were put 3 in a group for treatments with the various concentrations of the aqueous extracts of pawpaw parts and the control commercial coccidiostat, Atlacox (Atlas Pharma Co. Limited, UK). There were two replicates of each group for the treatments.
2. Culture and inoculation of coccidia
An intestinal lysate of chicken affected by coccidiosis was put in 2% solution of potassium bichromate solution in petri dishes. Five days were allowed for the coccidia oocysts to sporulate in the 2% potassium bichromate at 250C and 1ml this was orally administered to each of the experimental chicken. A period of 10 days was allowed for the establishment of coccidia in the chicken before treatments with the aqueous extracts the pawpaw parts.
4. Preparation of aqueous extracts of Carica papaya parts
The seeds, leaves, and roots of pawpaw were harvested, oven-dried at 50°C, pulverised with a hammer mill, put in a sealed polythene bags and kept in a fridge at 4°C until use. For the seeds and roots, an initial 25% concentration was prepared by addition of drinking water and left to stand for 24 hr after which the supernatant was decanted and serial dilutions made thereof also with drinking water. For the dried pulverized leaves the initial concentration was 50% because the leaves required so much drinking water to soak before one could squeeze out some liquid to serve as the supernatant. The following serial percentage concentrations were prepared with drinking water from the supernatant just before administration to infected chicken: 50%, 25 %, 12.5 %, 6.25 % and 3.125 %.
5. Treatments
To the individual birds in the categories of the experimental design, 2 ml of the various concentrations of aqueous extracts of the Carica papaya were administered orally each day for the 8-day experimental period by means of a syringe. One control group of birds was allowed to drink an aqueous solution of the commercial coccidiostat, Atlacox (0.025 %) ad libitum. The other control group was given only drinking water without any pawpaw extract (placebo).
6. Enumeration of coccidia oocysts
The total daily faecal output of the birds in each group was harvested for counting of coccidia oocyst by the modified McMaster technique. Essentially 3g of faeces was mixed thoroughly with 42 ml of tap water and strained with a wire mesh of aperture 0.15m. The filtrate was thoroughly mixed and an aliquot of the uniform suspension of faecal material centrifuged for two minutes at 1500 rpm. The packed sediment was emulsified. Saturated salt solution (NaCl) was added until the volume equaled that of the initial aliquot of filtrate. The tube was inverted several times until the sediment was evenly suspended and two chambers of McMaster slide were filled with the filtrate for counting (Manual of Veterinary Parasitological Laboratory Techniques).
Results and Discussion
After the 10 days allowed for the coccidia to establish in the experimental chicken an average of 15 950 oocysts/gm of faeces was counted from their pooled faeces. The experimental birds showed the symptoms of coccidiosis - visibly drowsy and brown near-watery faeces. Hall (1994) reported that mere presence of Eimeria oocysts in the faeces does not confirm that the disease is present because large numbers of oocysts may be present without causing clinical coccidiosis. North and Bell (1990) noted that the presence of, at least, 10 000 specific Eimeria species is required to show the signs and symptoms of coccidiosis. Clinical symptoms of the disease were observed before the start of the interventions of this study because 15 950 oocysts had been established. Oocyte count from birds on the water (placebo) was discontinued because they became too sick and a radical intervention was necessary to save their lives. None of the birds died during the trial period.
From Fig. 1, one can deduce that the aqueous extract of Carica papaya roots reduced coccidia oocyst production. The higher the percentage concentration of the aqueous extract, the greater the reduction of oocyst production. Whereas the control commercial coccidiostat, Atlacox, reduced coccidian oocyst counts by 1.8 logs10 gm-1 of faeces (98.4%) in 8 days, the 25% extract reduced the oocysts by 2.03 logs10 gm-1 (98.8%) of faeces during the same period. The concentration of the root extract of Carica papaya at which the reduction of oocyst production was approximately equal to that of the commercial coccidiostat was on the 8th day was 12.5%. This concentration (12.5%), therefore, becomes the 'concentration of choice' over the 25% aqueous extract of pawpaw roots because of Minimum Inhibitory Concentration (MIC) considerations.
Fig. 2 also suggests that an aqueous extract of the leaves of the pawpaw reduced coccidia oocyst production within the experimental period. The control commercial coccidiostat reduced oocyst counts by 1.8 logs10 gm-1 of faeces (98.4%) approximately within the experimental period. Paradoxically, however, it was rather the least concentration of the extract of the leaves of the pawpaw (6.25%) gave the greatest reduction of oocyst counts within the experimental period (2.33 logs10 gm-1 of faeces i.e. 99.5%) over the experimental period, making it a 'concentration of choice' for administration considering MIC.
Fig. 3 shows that the various percentage concentrations of aqueous extracts of pawpaw seeds were all stronger in reducing coccidia oocyte production compared to the control coccidiostat, Atlacox. Using 12.5% and 6.25% concentration of the aqueous pawpaw seed extract, no coccidia oocysts were detected by the 4th day. The oocyst populations had been reduced by 4.2 logs10 gm-1 (100%) of faeces by the 4th day. In comparison, the control coccidiostat caused a reduction of 1.44 logs10 gm-1 of faeces (96.3%) by the same 4thday. The 6.25% aqueous pawpaw seed extract will be, therefore, the 'concentration of choice' over 12.5% concentration considering MIC. Also within the 4-day period, the 'choice concentrations' of aqueous pawpaw leaves (6.25%) and roots (12.5%) extracts had reduced coccidia oocyst populations by 1.46 logs10 gm-1 and 1.63 logs10 gm-1 of faeces respectively.
Fig. 4 suggests that the 6.25% aqueous extract of the seeds of pawpaw has the greatest anticoccidia oocyst production effect compared to the most effective concentrations extracts of leaves (6.25%) and roots (12.5%) and even the control coccidiostat, Atlacox. Aqueous extracts of pawpaw seeds, leaves, Atlacox and root follow in that order considering the strength with which they inhibit coccidia oocyte production.
The general sharp reduction in oocyst numbers within the first few days after administration of all concentrations of the aqueous extracts and the Atlacox suggested an initial quick knockdown effect on coccidia oocyst production followed by a progressive, though slowed reduction in oocyst counts in subsequent days. North and Bell (1990) stated that for a drug to be effective it must remain at a high concentration in the body from 3-5 days. In this study, the dosage was same for each day and was theoretically high to effect the reduction in oocyst counts. It is suggested that the seemingly slowed effect of treatment thereafter is possibly due to interplay of resistance to the intervention and the developing immune status of the bird.
At the concentrations used and the time period of the experiment, no lesions were found that necessitated the rejection of the meat of the experimental birds as unwholesome.
Conclusion
Water extracts of root, leaves and seeds of Carica papaya have anticoccidia potential. The effectiveness is linked to the concentration of the extracts and the part of the plant from which the extract is prepared. Seeds, leaves and roots follow in that order according to their potency. This finding justifies the practice of giving pawpaw seeds to chicken in rural settings to alleviate coccidiosis. The implication of this finding is that in a rural setting in a developing country where a veterinarian is not readily available, aqueous solutions at the concentrations of this study may be used orally as a remedy to for coccidiosis. The concentrations of the aqueous extractions used in this study and during the experimental period did not show visible postmortem lesions to warrant the rejection of the meat the experimental birds as unwholesome. However, pathohistological investigation after long administration of these extracts, especially also of polar extracts, is necessary to determine toxicity at the cellular level. Further, identification of the chemical principle of Carica papaya seeds, root and leaves that accounted for the suggested anticoccidia effect needs to be identified for future advantage to be taken of it. Then dosage trials taking into consideration the age and species of the plant and the age of birds may be also necessary.
Acknowledgement
Funding for this work came from the general research allowances grated to the University of Cape Coast from the Ghana Government.
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Author Resource:-
Annan-Praha* A, A. A. Mensaha, S. Koduaa and M. A. Bentila
School of Agriculture, University of Cape Coast, Cape Coast, Ghana.
*Correspondence:
Dr. A. Annan-Prah
School of Agriculture
University of Cape Coast
Cape Coast
Ghana
Tel. + 233 20 8149333, fax +233 42 33842
E-mail annanprah@yahoo.com
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