A Study of the Potential of Using the Goat Weed (Ageratum Conyzoides) as an Anticoccidia Agent in Chicken

Journal of Veterinary Advances A Study of the Potential of Using the Goat Weed (Ageratum Conyzoides) as an Anticoccidia Agent in Chicken Annan-Prah A., Dayie N. T. K. D., Hackman S. M., Amporful A. J. J Vet Adv 2012, 2(7): 292-297 Online version is available on: www.grjournals.com

ISSN: 2251-7685 ANNAN-PRAH ET AL. A Study of the Potential of Using the Goat Weed (Ageratum Conyzoides) as an Anticoccidia Agent in Chicken Original Article Annan-Prah A., Dayie N. T. K. D., Hackman S. M., Amporful A. J. School of Agriculture, University of Cape Coast, Ghana Abstract This study investigated the speculation that the goat weed, Ageratum conyzoides, has antiprotozoan activity. Various concentrations of potable water and whole extracts of the leaves of Ageratum conyzoides were administered to 5-week old broiler chicken that had been infected with sporulated oocysts of Eimeria. The commercial coccidiostats, Amprox and Coxisol, were also administered for comparison. Faeces were harvested from the recta of the experimental chicken over a 5-day period and assayed for oocyst populations. Results of this study suggested that although an inoculum size of 7300 oocysts was administered to the experimental chicken, varying populations of oocysts (n) got established after 5 days before commencement of treatments. Pertinent coccidia populations reductions were as follows: 10%water extract 96% (n=3388), 10% whole extract 81.7% (n=1500), Coxisol 97% (n=1216) and Amprocox 85.9% (n=1600). Pathohistological investigation with treatment with 10% whole extract of Ageratum conyzoides did not find any lesions in the heart, liver, lungs kidney and spleen. Key words: Goat weed, ageratum conyzoides, coccidiosis, chicken Corresponding author: School of Agriculture, University of Cape Coast, Ghana Received on: 09 May 2012 Revised on: 13 May 2012 Accepted on: 20 Jul 2012 Online Published on: 30 Jul 2012 292 J. Vet. Adv., 2012, 2(7):292-297

A STUDY OF THE POTENTIAL OF USING THE GOAT WEED Introduction In remote areas of developing countries, veterinary services and orthodox drugs are either unavailable or unaffordable. Farmers, therefore, rely on herbal plants to prevent and cure many ailments. One such plant is the goat weed, Ageratum conyzoides. Burkill (1985) has described this plant as an athropogene because it is common in the proximity of human habitation. The plant has been evidenced to cure amoebesis, trypanosomiasis, sleeping sickness, crawcraw, skin infections, chronic ulcers, wounds, haemorrhoids, uterine bleeding, rheumatism, gastrointestinal disorders, eye troubles, otitis, gonnorrhoea, cough, pneumonia and jaundice (Watt and Breyer-Brandwijk, 1962; Altschul, 1973; Burkill, 1985; Oliver-Bever, 1986; Abbiw, 1990). The suggested uses are possibly due individual and/or collective actions of the plant s chemical components: phenols, eugenol, ageratochromone, chromenes, trierpenoids including steroids, flavenoids, conyzorium, methexnebilitin, quercetin, kaempferoglycosides, hydrocyanic acid (Oliver-Bever, 1986; Watt and Breyer-Brandwick, 1962; Ampofo, 1992), 6- dimethoxyageratochromone, ageratochromone (Katturi and Manithomas,1967; Rudolf, 1969), heptamethoxychromene, stigmasterol dotrioconthene, methoxydimethylchromene, conyzorigum (Adesogan and Okunade, 1979) and methoxynobilitin (Durodola, 1977). The suggested activity of Ageratum conyzoides against amoebesis and trypanosomiasis (both protozoans) makes one speculate that the plant can have a static or cidal effect on Eimeria, protozoans that cause coccidiosis, a disease that causes great morbidity and mortality especially in intensively reared chicken (Voeten, 1987, Kesse, 1988; McDougald, 1993). Among the most common pathogenic Eimeria for poultry are E. tenella, E, necatrix, E. brunetti, E mivati, E. maxima, E. acevulina, E. mitis, E. hagani and E. precox (, Lapage, 1968, Bruner & Gallispie, 1973; Hall, 1985). The first really effective anticoccidials with both preventive and curative properties were the sulphonamides, though they suffered from incomplete spectrum of activity against Eimeria (Brander et al., 1977). Eimeria have been developing continual resistance to industrially produced drugs (Chapman, 1993) and this has stimulated search for new methods of control (Allen et al., 1997). A. conyzoides could be one of the herbal remedies to combat the disease. The aims of this investigation were to study the effect of the administration of the following forms A. conyzoides on the survival of Eimeria oocysts in chicken: varying concentrations of whole extract, water extracts, dried pulverized leaves and chopped fresh leaves in feed. These were compared with industrially produced coccidiostats Coxisol and Amprocox (MARIDAV-GHANA under the license of RADAR, NV) and plain tap water. Materials and Methods Matured plants of A. conyzoides were harvested whole. One part of the fresh leaves was chopped and mixed with feed at a rate of 10%. Another part of the fresh leaves was oven-dried at 50 o C for 5 days to constant weight, pulverized by milling and mixed with feed also at the rate of 10%. Crude water extracts of 10% and 20% concentration were prepared by leaving 10g and 20g respectively of the dried pulverized leaves of A. conyzoides in 100 ml in distilled water overnight. The supernatant was decanted, filtered and stored at 4 0 C until their use for treatments. Whole extract was prepared by combining extracted non-polar, medium polar and polar components of the leaves of A. conyzoides. Extraction of polar components was done by vigorously shaking 356g dried, pulverised leaves overnight in 2 litres of 1:1 mixture of dichloromethane-methanol. The resulting extract was filtered from the mixture by suction pressure (Extract 1). The residue was shaken in 1000 ml methanol overnight and the resulting extract filtered by suction pressure (Extract 2). Extracts 1 and 2 were combined and evaporated to dryness. 1 gm of dried leaves gave 0.123 g of whole extract. Coccidia were cultured from the caeca content of birds with confirmed coccidiosis. The caeca contents were placed in 2% potassium dichromate solution in Petri dishes and coccidia oocysts in the caecal contents allowed to sporulate at 25 0 C for 5 days. The suspension was transferred into 120 ml conical flask and diluted with distilled water. The J. Vet. Adv., 2012, 2(7):292-297 293

ANNAN-PRAH ET AL. quantity of sporulated oocysts in 1 ml this was determined using the modified McMaster method (Manual of Veterinary Parasitological Laboratory Techniques, 1979). It was ascertained that each ml of the suspension which was administered to the experimental chicken contained 7300 sporulated oocysts. From a flock of 1,500 five-week old Lohman Brown chicken, 210 were randomly selected for the experiment. These chickens had undergone routine vaccinations (Marek, New Castle, Fowl Pox and Gumboro). They were fed on commercial feed except for the treatments that A. conyzoides had to be part of the ration. Water was given ad libitum. Fourteen groups of birds having 5 birds each with 3 replicates were formed for treatments. These were designated T1 T14. Each bird in every group was orally given 1 ml of the suspension of sporulated oocysts (ie. 7300). Five days were allowed for the coccidia to establish in the experimental chicken. Thereafter, treatments were as follows: T1 (Coxisol), T2 (Amprocox), T3 (2% whole extract of A. conyzoides), T4 (4% whole extract of A. conyzoides), T5 (5% whole extract of A. conyzoides), T6 (6% whole extract of A. conyzoides), T7 (7% extract whole of A. conyzoides), T8 (8% whole extract of A. conyzoides), T9 (10% whole extract of A. conyzoides), T10 (10% dry leaves of A. conyzoides in feed), T11 (20% fresh leaves of A. conyzoides in feed), T12 (10% water extract of A. conyzoides), T13 (20% water extract of A. conyzoides), T14 (water placebo). Coxisol and Amprocox, used in T1 and T2 were commercial coccidiostats, sold in Ghana by MARIDAV-GHANA under the license of RADAR, NV). The experimental period was 5 days. Faeces were collected daily from the cloaca of the chicken. The total daily oocyst output was enumerated by the modified McMaster method. After the experimental period the birds were slaughtered and the following parts taken and prepared for pathohistological investigation by the standard histological methods: heart, liver, lungs kidney and spleen. Results and Discussion Table 1 shows that although the experimental chicken received the same inoculum size of oocysts, different percentages of oocysts were established after 5 days. It is suggested that some oocysts in the inoculum may not have sporulated at all or were not viable to multiply in the intestines of the experimental chicken. There was also the possibility of the interplay of the immune status of the birds, which might have impacted on oocyst establishment and subsequent multiplication in the intestine of the experimental birds. The number of coccidia oocysts/g of faeces at the end of the 5 th day after the initial administered inoculum size of 7300 was taken as a reference number or percentage (100%) from which oocysts production was to be counted. Table 2 shows a general decrease of oocysts numbers/ gm for the treatments. Coxisol, having reduced oocysts population by 97% was seemingly better than Amprocox (85.9%) as a commercial coccidiostat. Although all treatments with whole extracts of A. conyzoides reduced coccidia populations, none superseded Coxisol or Amprocox. The higher the percentage of administered whole extract of A. conyzoides, the greater was the reduction, the highest being 10% of whole extract of A. conyzoides. This concentration of whole extract (10%) seemed to have reduced the coccidia oocysts population to levels that Amprocox did after 5 days, though the established coccidia numbers on the first day of treatment with 10% A. conyzoides was 100 oocysts lower. We should, however, not lose sight of the fact that the effectiveness of a cidal agent also depends on the initial population size. We had noted in Table 1 that despite the same inoculum size the number of established oocysts varied. From Table 3, 10% water extract of A. conyzoides (T12) caused more reduction of oocysts numbers in the experimental period than 20% water extract (T13) did, though the initial oocyst counts at the beginning of the experiment was higher for 10% water extracts (T12) than for 20% water (T13). The oocysts reduction for 10% water extract (T12) was comparable to T1 (the commercial coccidiostats, Coxisol). Since the initial established number of oocysts was higher for the 10% water extract of A. conyzoides (T12), it gives the indication that 10% water extract of A. conyzoides was stronger than 294 J. Vet. Adv., 2012, 2(7):292-297

A STUDY OF THE POTENTIAL OF USING THE GOAT WEED Coxisol in reduction of oocysts (but not necessarily less toxic after long application). Table 1: Coccidia establishment in chicken 5 days after inoculation (oocysts/ gm of faeces) Treatment Initial Coccidia established % coccidia established inoculum after 5 days after 5 days T1 (Coxisol) 7300 1216 16.1 T2 (Amprocox) 7300 1600 21.9 T3 (2% whole extract of A. conyzoides ) 7300 1850 25.3 T4 (4%whole extract of A. conyzoides) 7300 2100 28.7 T5 (5%whole extract of A. conyzoides) 7300 1250 17.1 T6 (6%)whole extract of A. conyzoides ) 7300 1600 21.9 T7 (7% whole extract of A. conyzoides) 7300 1100 15.1 T8 (8%whole extract of A. conyzoides) 7300 1375 18.8 T9 (10%whole extract of A. conyzoides) 7300 1500 20.5 T10 (10% dry leaves in feed) 7300 2250 30.8 T 11(20% fresh leaves in feed) 7300 663 9.1 T12 (10% water extract) 7300 3389 46.4 T13 (20% water extract) 7300 2876 39.3 T 14 ( water placebo) 7300 363 4.9 Table 2: Coccidia reduction during experimental period Treatment D0 D1 D2 D3 D4 D5 T1 (Coxisol) 1216 (100) 239 (80.3) 55 (95.5) 69 (94.5) 79 (93.5) 36 (97) T2 (Amprocox) 1600 (100) 1300 (18.6) 775 (51.6) 350 (78.1) 273 (82.9) 225 (85.9) T3 (2% whole extract A. conyzoides) 1850 (100) 1650 (10.8) 1075 (41.9) 1375 (25.7) 1300 (29.7) 500 (85.9) T4 (4%whole extract A. conyzoides) 2100 (100) 1725 (17.9) 1125 (46.4) 425 (79.8) 250 (88.1) 675 (67.9) T5 (5%whole extract A. conyzoides) 1250 (100) 1750 (-40) 1600 (28) 450 (64) 300 (76) 500 (60) T6 (6%whole extract A. conyzoides) 1600 (100) 1725 (-7.8) 1500 (6.3) 975 (39.1) 675 (57.8) 300 (81.5) T7 (7%whole extract A. conyzoides) 1100 (100) 950 (13.6) 1100 (0) 475 (56.8) 375 (65.9) 325 (70.1) T8 (8%whole extract A. conyzoides) 1375(100) 925 (32.7) 775 (43.6) 635 (56.8) 550 (60) 675 (50.9) T9 (10%whole extract A. conyzoides) 1500 (100) 1000 (33.3) 625 (58.3) 325 (78.3) 325 (78.3) 275 (81.7) T 14 ( water - placebo) 363 (100) 376 (-3.6) 401 (-10) 548 (-50) 1598 (- 340) 3506 (-865) * Percentages in parenthesis Table 3: Comparison of oocysts reduction by commercial coccidiostats, A.conyzoides leaves in feed and its water extract Treatment D0 D1 D2 D3 D4 D5 T1 (Coxisol) 1216 (100) 239 (80.3) 55 (95.5) 69 (94.5) 79 (93.5) 36 (97) T2 (Amprocox) 1600 (100) 1300 (18.6) 775 (51.6) 350 (78.1) 273 (82.9) 225 (85.9) T9 (A. conyzoides 10%) 1500 (100) 1000 (33.3) 625 (58.3) 325 (78.3) 325 (78.3) 275 (81.7) T11 10% dried leaves in feed 2928 (100) 672 (77) 3306 (-12.9) 2189 (25.2) 2639 (9.8) 4616 (-57) T12 10% fresh leaves in feed 863 (100) 450 (51.3) 271 (68.6) 742 (14) 2692 (-211.9) 1717 (-98) 10% water extract 3388 (100) 735 (78.3) 248 (92.6) 202 (94) 212 (93.7) 125 (96.3) 20% water extract 2877 (100) 907 (68.5) 197 (93.2) 255 (91.1) 520 (81.9) 504 (82.4) T 14 ( water placebo) 363 (100) 376 (-3.6) 401 (-10) 548 (-50) 1598 (-340) 3506 (-865) * Percentages in parenthesis For water treatment (control), the increasing number of oocysts could be expected or speculated. That, A. conyzoides in water (T14) reduced numbers of coccidia oocysts, when plain water did not, make the suspicion of an anticoccidia principle in A. conyzoides stronger. Leaves of A. conyzoides, whether incorporated dried or fresh in feed, did not seem to have had any impact on oocysts reduction. J. Vet. Adv., 2012, 2(7):292-297 295

ANNAN-PRAH ET AL. Fig. 1: Histological section of heart after 5day treatment with 10% Ageratum conyzoides. Fig. 4: Histological section of lung after 5day treatment with 10% Ageratum conyzoides. Fig. 2: Histological section of liver after 5day treatment with 10% Ageratum conyzoides. Fig. 5: Histological section of spleen after 5day treatment with 10% Ageratum conyzoides. In both cases, oocysts counts rather rose. Histological pictures of the heart, liver, lungs kidney and spleen that had been treated with 10% whole extract of A. conyzoides did not show any pathological lesions within the 5-day period of the study and at the rates of application. It is worthy of note, however, that Trigo et al. (1988) found that A. conyzoides contain 1,2-desifropirrolizidin and licopsamine which can have hepatotoxic activity. Conclusions Fig. 3: Histological section of kidney after 5day treatment with 10% Ageratum conyzoides. There is an anticoccidial principle in A. conyzoides that needs to be identified and exploited. The dosage and long-term toxic effect of treatment with A. conyzoides needs further studies. For the ordinary farmer in the rural areas in developing 296 J. Vet. Adv., 2012, 2(7):292-297

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