Efficacy of Piperazine citrate, Levamisole hydrochloride and Albendazole in the treatment of chicken naturally infected with gastrointestinal helminths

H W Chege, D C Kemboi, L C Bebora, N Maingi, P G Mbuthia, P N Nyaga and L W Njagi

University of Nairobi PO Box 29053-00625 Nairobi, Kenya
wambuiche08@gmail.com

Abstract

A controlled study was carried out in the Department of Veterinary Pathology, Microbiology and Parasitology, Kabete campus, University of Nairobi, Kenya during the month of March 2012 to determine the comparative efficacy of, piperazine citrate at a dose of 3mg/kg body weight (BW), levamisole hydrochloride at a dose of 25mg/kg body weight, and albendazole at a dose of 20mg/kg BW. Thirty seven adult normal feathered female chicken were purchased from individual farmers in Mbeere Sub county, for this experiment. The chicken were divided into 4 groups. Chicken in groups 1, 2, and 3 were treated with piperazine citrate, levamisole HCL and albendazole, respectively, while group 4 chicken were kept as untreated controls. Albendazole was administered orally as a single dose while piperazine citrate and levamisole HCL were given for 24 hours in drinking water. Post -mortem examination for parasites was done twice: (1) when birds were sacrificed before start of the experiment, to ascertain presence and types of parasites in the birds, and (2) when the experimental birds were sacrificed 7 days post treatment (end of experiment). Throughout the experimental period, birds were kept separately in cages where faecal samples were collected at intervals and screened for parasite eggs.

Birds that were examined at start of experiment were found to harbour nematodes (mainly ceacal worms), cestodes (mainly Raillietina tetragona and Raillietina echinobothrida) and Tetrameres americana. Piperazine citrate was effective against ascarids (which were found in one bird only) but had no effects on other worms. Levamisole HCL was 100% effective against the caecal worms. It had very little efficacy of 25.6% and 17.6% against cestodes Raillietina echinobothrida and R. tetragona respectively and 62.8% efficacy against Tetrameres americana. Albendazole was 100% effective against the caecal worms and the cestodes (Raillietina echinobothrida and R. tetragona) and Tetrameres americana. Results indicated that Albendazole at 20mg/kg BW was the most effective with respect to treatment of cestodes, roundworms (and ascarids). The use of albendazole is, therefore, recommended to ensure total control of worms.

Key words: anthelmintics, indigenous chicken, Kenya, parasites

Introduction

Endo- and ecto- parasites are common among indigenous chicken since they are kept outdoors where they scavenge and forage and in the process pick up the infective stages of the parasites. These parasites compete with the birds for nutrients; some suck blood causing anaemia, while others cause anorexia or death. Some ecto-parasites are important in transmission of certain pathogens, while others may cause disease such as scaly leg and depluming mange.

Anthelmintic intervention often involves medication with piperazine, tetramisole and oxfendazole. However these anthelmintics generally exhibit low efficacy and are associated with undesirable effects (Verma et al 1991). A study conducted in Kansas State University, USA, to evaluate the efficacy of levamisole in drinking water against some nematodes of chicken, demonstrated that at a dose of 48 mg/kg body weight was 100% efficacious against Heterakis gallinarum. It was also found that levamisole given in drinking water to chicken at dosages of 18 or 24 mg/kg body weight was 100% efficacious against Ascaridia galli (Cruthers et al 1975).  No such studies have been done in Kenya; this is the first one.

Considering the fact that heavy parasite burdens have been recorded in indigenous chicken in Eastern Province of Kenya (Sabuni 2009), it was found necessary to determine the effectiveness of various anthelmintics that can be used. The study was, therefore, carried out to address the identified gaps as a step towards improving the control of endo-parasites.

Materials and methods

Experimental birds

Thirty seven normal feathered adult female (> 8months) indigenous chicken managed entirely by free range manner were purchased from individual farmers in Mbeere Sub-county.  The ages of the birds were determined based on the size of the crown, length of the spur, flexibility of xiphoid cartilage and information from the poultry farmers as previously done by Magwisha et al  (2002) and Sabuni (2009). Adult females were selected on this the basis that they were readily available and could stay longer than the cockrel which are normally eaten earlier.

The chickens were transported alive in cages to the Department of Veterinary pathology, Microbiology and Parasitology. The chicken were allowed to acclimatize for three days at the research facility under housing, maintenance and feeding until the whole experiment was over.            

Experimental design

On day 0, each individual bird was caged differently (Figure 1)   

Figure 1: Chicken caged separately, faecal pots, feeding and watering troughs during the experiment

Before start of experiment, post-mortem examination was done on 7 birds to determine the type of endo-parasites the birds were carrying.

No pre-trial treaments were done , because this experiment  aimed to evaluate  the efficacy of   anthelmintic  readily available in the market and what the farmers were using in the study area.

 On day 4 the chicken were randomly allocated to the treatment groups; however, the only bird that excreted ascarid eggs was purposely placed in the piperazine treatment group. There were 4 treatment groups with 9 chicken for control, 7 chicken for albendazole, 7 chicken for levamisole HCL and 7 chicken for piperazine citrate (N compatible with World Association for the Advancement of Veterinary Parasitology guidelines that require a minimum of 6 infected birds per treatment group on day 0 (Yazwinski et al 2003) (Table 1).

Treatment regimes

Albendazole was used at a rate of 20 mg/kg of body weight (BW). The highest weight of the bird was used (2.5kgs) in calculating the volume (0.5mls) of albendazole used. The drug was administered orally as a single dose (According to the manufacturer’s instructions).

Levamisole was given 25 mg/kg body weight (7.5ml) put in three litres of water. Equal division of the medicated water was done among the 7 birds where each bird was given 430millilitres (ml) of water (According to the manufacturer’s instructions). After 24 hours the amount of medicated water left was measured to ascertain the amount of water taken by the chicken.

 Three quarter tea spoonful of piperazine citrate (4.5gm) was dissolved   in three litres of water for 7 birds. Each bird was given at 3 mg/kg body weight derived using the highest weight of the bird. Equal division of the medicated water was done among the 7 birds where each bird was given 430 mls of water (According to the manufacturer’s instructions). After 24 hours the amount of medicated water left was measured.

Faecal egg count and identification

Daily faecal samples were collected from the cage three times a day; morning, noon and evening. The faecal samples were examined for nematode egg counts using the modified McMaster technique (MAFF 1986).

 Parasite recovery and determination of effectiveness of anthelmintics

 Procedures used for parasite recovery were according to the WAAVP guidelines for evaluation of anthelmintics in poultry (Yazwinski et al 2003) as described below.

Efficacy of anthelmintics was evaluated by

a)      Complete reduction of parasite eggs in the treated chicken

b)      Percentage effectiveness against each parasite species (or stage) was determined using the formula of Yazwinski et al (2003).

Key: % E = Percentage effectiveness, No. = number

Seven (7) days after treatment the birds were sacrificed and the parasites recovered, identified and counted (MAFF 1986).

The means of helminth population for each treatment group were used to calculate the percentage efficacy of the anthelmintics. Percentage efficacies for the different anthelmintics were considered effective above 90 %.

Results

The daily faecal egg count  of  the 30 chicken  used in this prior to treatment  showed  two chicken were positive for Heterakis species eggs and one for Ascaridia galli eggs (Figure 2).

Figure 2: Heterakis species (H) and Ascaridia galli eggs (A) isolated from chicken faecal sample

The chicken shed the eggs more in the morning than in the noon and evening. The shedding of the eggs was completely reduced two days later after the treatment.

After treatment no adverse effects were observed on birds’ appearance, behaviour and appetite. The helminths found at necropsy after treatment were the nematodes (caecal worms and Tetrameres americana) and cestodes (Raillietina echinobothrida and R. tetragona).

Table 2 shows the percentage efficacies for different anthelmintics used.

Albendazole at 20 mg/kg body weight was 100% effective against Heterakis species, H. isolonche, Subulura brumpti, Tetrameres americana, Raillietina tetragona and Raillietina echnobothrida. Albendazole at 20 mg/kg body weight was the most effective in all the treatments carried out for these helminths.

Figure 3 shows distorted tapeworm segment after treatment with Albendazole at 20 mg/kg body weight.

Figure 3: Distorted tapeworm segment (white arrow) after treatment with Albendazole

Levamisole HCL 25 mg/kg was 100% effective against the caecal worms. It had very little efficacy of 25.6% and 17.6% against cestodes Raillietina echnobothrida and R. tetragona respectively and 62.8% efficacy against Tetrameres americana.

Piperazine citrate was not effective against cestodes (Raillietina species), caecal worms Heterakis species, Subulura brumpti) and Tetrameres americana. But it was observed that it was effective against Ascaridia galli which was only present in one of the experimental bird.

Discussion

Results of this study indicated that the chicken were infected with caecal worms and the cestodes Raillietina tetragona and Raillietina echinobothrida. Piperazine citrate at 3 mg/kg body weight was only effective against Ascaridia galli and not effective against cestodes and nematodes contrary to the farmers’ belief. From oral interview with the farmers in Mbeere, they indicated wide usage of piperazine on their birds; in fact, the farmers seemed to take Ascarex^R/Piperazine citrate as the only commercial anthelmintic (Chege et al 2014). This could thus be the reason for not recovering Ascaridia galli from the birds – Piperazine citrate could have eliminated them. The fact that Piperazine citrate had no effect on other parasites explains why carriage of the other parasites in the birds was high.

 With respect to other parasites, Levamisole HCL at a dose of 25 mg/kg body weight was only effective against the caecal worms; Heterakis species, Heterakis isolonche and Subulura brumpti. It had no effect on tapeworms and Tetrameres americana. Other studies in Sudan (Thienpoint et al 1966), found that Levamisole HCL did not have any anthelmintic efficacy against tapeworms. Most of the farmers in the area are not familiar with Levamisole HCL which is commercially available and did not therefore use the drug. Levamisole HCL poultry formulation in liquid form is commercially readily available in Kenya.

Albendazole at 20 mg/kg body weight was 100% effective against Heterakis species, Subulura brumpti, Tetrameres americana, Raillietina tetragona and Raillietina echinobothrida. Other studies in Arkansas, United States (Tucker et al 2007) reported that Albendazole at 20 mg/kg body weight was effective against Ascaridia galli, Capillaria obsignata, Heterakis gallinarum and Raillietina cesticillus but they did not report on Subulura brumpti, Tetrameres and the two species of tapeworms reported in this study. Albendazole was also effective in deworming chicken infected with R. cesticillus (96.2% reduction) and caused no adverse effects (Tucker et al 2007). A similar study in Sudan (Saeed 2007) showed that albendazole administered at 25 mg/kg body weight was 100% effective against experimental Raillietina tetragona infection in chicken.

This is the first experiment of this kind to be done on chicken and reported in Mbeere Sub-county, Kenya. 5% Albendazole powder form for poultry is readily available in China but not in Kenya. Most pharmaceutical companies in Kenya are challenged by high cost of production of the drug.

In this study, no results are given, with respect to effectiveness of Levamisole HCL and Albendazole on ascarids, because the experimental chicken did not have the parasites. However, from literature (past experiments), the two are effective against ascarids.

Conclusions

• The most effective drug was Albendazole 20 mg/kg body weight - against cestodes (Raillietina echinobothrida and Raillietina tetragona) and nematodes (Caecal worms and Tetrameres americana.

  
  

• Levamisole HCL was 100% effective against caecal worms (Heterakis species and Subulura brumpti). It was not effective against cestodes

  
  

• Piperazine citrate (Ascarex) was not effective against cestodes (Raillietina echinobothrida and R. tetragona) and nematodes (caecal worms and Tetramere americana) but it was effective against Ascaridia galli which was present in one bird.

  
  

• There was no correlation between the faecal egg count and the worm burden.

Recommendations

The use of albendazole is recommended to ensure total control of worms. Unfortunately there are no prescription drugs for treatment of avian cestodes; there is need to look into this.

Acknowledgements

The authors thank The Regional Universities Forum for Capacity Building in Agriculture (RUFORUM) for their sponsorship and funding of the project, the University of Nairobi for providing the work space  and facilities, Richard Otieno, Rose Nyawira, Rebecca Githinji and  Dorcas Nduati  for their technical assistance and the farmers of Mbeere for their cooperation.

References

Chege H W, Kemboi D C, Bebora L C, Maingi N, Nyaga P N, Mbuthia P G, Njagi L W and Githinji J 2014 Chicken parasites and local treatments used against them in Mbeere District, Kenya. Livestock Research for Rural Development Volume 26, Article #2. Retrieved August 13, 2014, from http://www.lrrd.org/lrrd26/1/cheg26002.htm

Cruthers L R, Al-Khateeb G H, and Hansen M F 1975 Efficacy of levamisole (Tramisol) in drinking water against some nematodes of chicken. Proceedings of Oklahoma Academy of Science, 55: 119-121.

Magwisha H B, Kassuku A A,  Kvysgaard N C and Permin A 2002  A comparison of the prevalence and burdens of helminth infections in growers and adult free-range chickens. Tropical Animal Health and Production, 34: 205-214.

Ministry of Agriculture, Fisheries and Food (MAFF) 1986 Manual of Veterinary Parasitological Techniques. Technical Bulletin. No 18. HSMQ. London. pp. 118-124.

Sabuni A Z 2009 Prevalence, intensity and pathology of ecto- and haemo-parasites infections in indigenous chicken in Eastern province of Kenya. MSc thesis. University of Nairobi.

Saeed A M 2007 Efficacy of albendazole against experimental Raillietina tetragona infection in chickens. Research Journal of Pharmacology, 1: 5-8.

Thienpoint D, Vanparijs O F S, Raeymaekers A H M, Allewijn F T N, Marsboom R  P L, Niemeggeers C J E, Janssen K H  L, and Tetramisole (R829a) 1966 A new potent broad spectrum anthelmintics. Nature, 209: 1084-1086.

Tucker C A, Yazwinski T A, Reynolds L, Johnson Z and Keating M 2007 Determination of the anthelmintic Efficacy of Albendazole in the Treatment of Chickens Naturally Infected with Gastrointestinal Helminths. Journal of applied poultry research 16: 392-396.

Verma N, Bhatnager P K  and Banerjee D P 1991 Comparative efficacy of three broad-spectrum anthelmintics against Ascaridia galli in poultry. Indian Journal of Animal science, 61: 834-835.

Yazwinski T A, Chapman H D, Davis R B, Letonja T, Pole L, Maes L, Vercruysse J and Jacobs D E 2003 World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines for evaluating the effectiveness of anthelmintics in chickens and turkeys. Veterinary Parasitology, 116: 159-173.

Received 5 March 2017; Accepted 20 March 2017; Published 1 May 2017

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