The anthelmintic efficacy of ivermectin, levamisole and albendazole against strongyle and tapeworm infections in sheep

doi:10.21203/rs.3.rs-2608332/v1

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The anthelmintic efficacy of ivermectin, levamisole and albendazole against strongyle and tapeworm infections in sheep

https://doi.org/10.21203/rs.3.rs-2608332/v1

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The biggest obstacles to the welfare and productivity of small ruminants worldwide are gastrointestinal tract parasite infections. These parasites have been controlled largely by chemical anthelmintics. However, these parasites have developed resistance to some of these anthelmintics. Unfortunately, Ghanaians are not aware of the exact level of resistance that these parasites have developed against the known anthelmintics. This study was conducted to determine the efficacies of albendazole, ivermectin and levamisole against strongyle and tapeworm infections in Sheep.

Sixty (60) sheep were chosen at random and split equally into four groups: the first group received albendazole, the second group received ivermectin, the third group received levamisole, and the fourth group received distilled water. Faecal samples were directly collected from the rectum of each animal at Pre-treatment (Day0) and post-treatment (Day7 and Day14). The faecal samples were conveyed to the Spanish laboratory of the University for Development Studies, and analyzed using the modified McMaster method. At Day14 post-treatment, the fecal egg count reduction test and cure rate were used to determine the effectiveness of the anthelmintics.

The efficacies of albendazole, ivermectin, and levamisole, were 86.2%, 89.8%, and 95.4%, respectively, against strongyles. Levamisole had 0% cure rate against tapeworm infection in sheep at day14 post-treatment. This study brought to light that, the strongyles in the study area are suspected of developing resistance to albendazole and ivermectin.

Anthelmintic resistance

efficacy

albendazole

Ivermectin

Levamisole

Sheep

Small ruminants perform many functions for the well-being of poor and landless households (Bettencourt et al., 2015). These animals are not only reared for socioeconomic gains, but also for monetary gains (Barratt et al., 2019) including manure, hide, source of medium-term savings, investment diversification, and social and cultural purposes among others (Weyori et al., 2018). In rural livelihoods, the animals also serve as insurance to cover unforeseen expenses such as school fees and medical bills (Verpoorten, 2009).

Small ruminant meat is a principal protein source in several local cereal-based meals, and it can help pregnant women and vulnerable children eat better (Horasanli, 2010). These animals also contribute significantly to the long-term viability of smallholding farming systems (Devendra and Chantalakhana, 2002). For example, animal excrement serves to improve on the fertility of the soil, saving farmers money on chemical fertilizer (Karbo et al., 1997).

Globally, livestock illnesses have a considerable economic impact. Gastrointestinal tract (GIT) helminth infections have been recognized as the most significant livestock illness in a worldwide assessment that categorized livestock disease relevance base on its impact on the poor. Grace et al. (2017) found that Sub-Saharan Africa is home to the majority of the world's impoverished livestock keepers. GIT parasite infections in ruminants have negative consequences on feed intake, carcass composition and weight, growth rate, wool growth, milk yield, fertility among others (Fitzpatrick, 2013).

Infections with endoparasites can be obtained by the consumption of helminth eggs or infective larvae found on contaminated food, plants, soil, water, infective cysts found in the tissue of the intermediate hosts such as arthropods and gastropods, transplacental or direct skin penetration transmissions (Greenland et al., 2015). In Ghana, many GIT parasites have been identified in small ruminants, with high prevalence rates in sheep (Squire et al., 2013; Owusu et al., 2016). In many regions of the world, there is the rise of anthelmintic resistance (Ram et al., 2007; Kumsa et al., 2010). Despite the availability of numerous control strategies, chemotherapy and chemoprophylaxis remain the most efficient means of combating parasitic infections. Some of the widely used anthelmintics available in Ghana are tetramisole (imidazothiazoles group), albendazole (Benzimidazoles group) and ivermectin (Macrocyclic lactones group), have been used for the last 40 years to treat and control gastrointestinal parasites infections in sheep.

To the best of our knowledge, no study has been conducted to determine the efficacies of albendazole, ivermectin and levamisole against gastrointestinal parasites in sheep in Ghana, hence the need to undertake this study. This would grant us the opportunity to determine which of the three drugs is the most efficacious against gastrointestinal parasites in Sheep.

Study area

The study was carried out in Wa East district, which is located in the Upper West Region of Ghana. Three settlement areas (Vissie, Kabanye and Kanbali) within the Wa East district were conveniently selected for the study. The district is found in the south eastern side of the the region. The Wa East district capital is Funsi, which is approximately 115km from the Upper West regional capital, Wa. The Wa East district shares boundaries with the West Gonja to southeast, to the North with Sissala East district and the West Mamprusi to the Northwest. The district has a total landmass of approximately 4297.1sq/ km², which is found between longitude 1º 10W and 2º 5’W and latitudes 9º 55’N and 10º 25’N. The climate in the district is tropical equatorial, which occurs across the northern part of Ghana. The temperatures are high all-year round, attaining its peak in March and April (GSS, 2010).

Study Population,design and sample size

The study population was all sheep not less than 4 months old. The sheep for the study were those reared under the semi-intensive system of production, whose owners were approached and permission granted for sample collection. A total of 60 sheep were used for the study, from the three selected areas within the study area.

Age categorization

Sheep with no permanent teeth were classified as young sheep while those who were having at least a pair of permanent teeth were classified as adult sheep.

Mode of application of treatment and dosage

Each sheep was then assigned randomly to one of the four groups (A, B, C and D). The sheep in group A were given albendazole 2.5% at a dose rate of 2ml/10kg body weight, those in group B were treated with ivermectin 1% at a dose rate of 1ml/50kg bodyweight and those in group C were given levamisole at a dose rate of 0.5ml/20kg body weight whiles those in group D were treated with 10ml distilled water per sheep. Animals in group A and D were given their interventions orally, the sheep in group B and group C were given their interventions Subcutaneous and intra-muscular routes respectively. All four treatments were given depending on the body weight of the animals, with the exception of the distilled water which was constant for each animal.

Faecal Samples Collection

Faecal samples were collected from each of the 60 sheep for the study, at pre-treatment (Day0) and post-treatment (Day7 and Day14). Each animal was given a unique tag before the pre-treatment for easy individual identification. Hand gloves were used and a small amount of water-based lubricant was applied to index and middle fingers. The middle and index fingers were then inserted into the rectum of each animal, and removed approximately, 4grams of fecal matter from them. The faecal matter from each sheep was then kept in a feacal sample container and well labeled for easy identification. The samples were then placed in ice chest with ice packs and transported to the Spanish Laboratory, University for Development Studies (UDS) for analysis using a McMaster technique.

Faecal Worm Eggs and Oocyte Count

A modified McMaster technique was used to analysis the feacal samples for the helminth load. About 3g of macerated faeces from each animal was kept in a beaker and then 42ml of distilled water was added to the faecal matter in the beaker and mixed. About 10ml of the suspension was then transferred into a 15ml test tube and centrifuged to 5000 rpm for 3 minutes at room temperature. The supernatant fluid was then poured off and the tube filled with about 3 ml saturated salt solution, which was then well mixed and centrifuged for a second time at 3,000 rpm for 3 minutes. The supernatant fluid was then sampled with the aid of a pasteur pipette and both chambers of the McMaster slides were filled. The filled McMaster slide was allowed to stand for 5 minutes to settle, and then placed under a light microscope. The counting chamber was then examined under the light microscope at a ×100 magnification. All the eggs found within the engraved areas of both chambers were counted and recorded. The total eggs within both chambers were then multiplied by 50 to obtain the egg count per gram of faeces.

Calculating the FECRT and Cure rate

The Strongyles effect of the various treatments were done using faecal egg count reduction test (FECRT) as contained in the equation below (Dash et al., 1988)

Where “T” and “C” are the arithmetic means in the treatment and control groups respectively while, 1 and 2 are pre-treatment and post-treatment respectively.

Whilst the efficacies of the treatments against tapeworm infections were done using the cure rate (CR) through the equation below.

Where “X’’ is the number of animals excreting tapeworm eggs at Day14 and “Y” is the number of animals excreting tapeworm eggs at pre-treatment (Day0).

Statistical Analysis

The data were captured into excel and imported into R version 4.2 for analysis. Descriptive statistics was computed for the means and standard errors of FEC with regards to the various treatment groups. A two-way ANOVA was conducted to determine the effect of the various treatments across the study period on the egg counts. Multiple comparison of means was done using a Student-Newman-Keuls (SNK) test at Day14 post-treatment. Statistical threshold of p < 0.05 was set. The results from the analyses were presented in tables and figures.

The strongyle FEC of the sheep treated with albendazole reduced from 680 ± 192.28 EPG at Pre-treatment to 133.33 ± 37.37 EPG at Day7 and 93.33 ± 28.4 EPG at Day14. The sheep treated with levamisole had their pre-treatment egg count of 613.33 ± 181.49 EPG reduced to 73.33 ± 30.03 EPG at Day7 and 26.67 ± 11.82 EPG at Day14. The sheep treated with ivermectin had their pre-treatment egg count of (626.67 ± 180.86 EPG reduced to 186.67 ± 55.09 EPG at Day7 and 80 ± 32,66 at Day14. The sheep treated with distilled water (Dw) had their pre-treatment egg count of 466.67 ± 152.34 EPG reduced to 453.33 ± 142.05 at Day7 and increased to 526.67 ± 148.15 at Day14 (Fig. 1).

The average tapeworm FEC of the sheep treated with albendazole reduced from 126.67 ± 57.29EPG at Pre-treatment to 30 ± 15.28EPG at Day7 and 0 ± 0EPG at Day14. The sheep treated with levamisole had their pre-treatment egg count of 66.67 ± 31.87 EPG increased to 70 ± 35.8 EPG at Day14. The sheep treated with ivermectin (IVER) had their pre-treatment egg count of 146.7 ± 72.9 EPG reduced to 3.33 ± 3.33EPG at Day14. The sheep treated with distilled water (Dw) had their pre-treatment egg count of 40 ± 21.38 EPG increased to 80 ± 36.77 at Day14 (Fig. 2).

Sheep given albendazole at pre-treatment had 40% tapeworm prevalence, which reduced to 27% at day7. The prevalence of tapeworm infection in the sheep given ivermectin at day0 and day14 were 33% and 7% respectively. The prevalence of tapeworm infections at day7 and day14 were higher than that of day0 (Fig. 3).

The strongyle egg count of the sheep given albendazole at pre-treatment in Kabanye was 740EPG. At same period, the sheep in Kanbali given distilled water was 560EPG. While, the average strongyle egg count in sheep given Ivermectin in Vissie at Day0 was 780EPG (Fig. 4).

At Day 14 after treatment, the FECRT brought to light that, albendazole had a mean efficacy of 86.2% with a lower 95% Cl of 76.8% against strongyle, while the drug produced an efficacy of 100% against tapeworm. The sheep in group D (those given levamisole) had their egg count reduced by 95.4% with a lower 95% Cl of 90.4% against strongyles when compared with their egg counts at pre-treatment (Day0). The efficacy of albendazole, ivermectin and levamisole against strongyle infections were statistically not different from each other (P = 0.222). However, albendazole, ivermectin and levamisole had a significant effect against tapeworm infections (P < 0.001). The World Association for the Advancement of Veterinary Parasitology (WAAVP), which explains anthelmintic resistance (AR) as a fecal egg count reduction (FECR) of less than 95% and a lower 95% confidence interval (CI) of less than 90%; if only one of these two factors is present, the farm is defined as being ‘suspected’ of having resistance. Efficacies of albendazole and ivermectin against strongyles were below 95% threshold set by WAAVP, hence, strongyles in the study area are suspected of having developed resistance to both ivermectin and albendazole, although the two anthelmintics (Albendazole and ivermectic) were effective (> 86%) against strongyles. Levamisole had 0% efficacy against tapeworm infection (Table 1).

Table 1

Efficacy of the various treatments against strongyles and tapeworm at day 14 post-treatment
Group	Treatments	Strongyles FECR (95% CI)	Tapeworm CR (95% CI)
A	Albendazole	86.21(76.82–95.6)	100^a
B	Ivermectin	89.81(83.17–96.45)	80^a(54.8-105.2)
C	Levamisole	95.39(90.39-100.39)	0^b(-28.1-28.1)
D	Distilled water -		-
P-value		0.222	<0.001
FECR = Faecal egg count reduction and CR = Cure rate

Various commercial anthelmintics such as albendazole, ivermectin, levamisole among others are currently being used for controlling of endoparasites in sheep in Ghana. However, the exact efficacies of these anthelmintics against helminths such as strongyles and tapeworms in Wa west district of Ghana is unknown. The study was therefore carried out to assess the efficacy of ivermectin, levamisole and albendazole against Strongyles and tapeworm infections in sheep.

The post-treatment strongyle EPG in sheep given albendazole, ivermectin and levamisole were lower than their corresponding EPG at pre-treatment. This shows the effect of those anthelmintics against strongyle in the Wa East district of Ghana.

The efficacy of ivermectin in the present study against strongyles is comparable with the 89% efficacy obtained by Adediran and Uwalaka (2015). The findings of the present study indicates that, levamisole had the highest efficacy against strongyles, while albendazole had the least efficacy against strongyles in sheep. This trend of efficacy of the three anthelmintics against strongyles is in agreement with the findings of Nsereko et al. (2013) and Teklemariam et al. (2016) against nematodes.

This may be due to the relatively non-frequent usage of levamisole compared to albendazole and ivermectin. Levamisole in sheep has been well known to cause adverse reactions such as anorexia, seizures, atexia paralysis of respiratory muscles (Hsu, 2008; Rehni & Singh, 2010) and frothy salivation particularly when over- dosed (Dar et al., 2020). Base on the adverse reactions in animals, levamisole is usually administered with caution (Dar et al., 2020), this may result in veterinary practitioners and farmers choosing those anthelmintics with relatively less adverse effects than levamisole. This may result in levamisole being less frequently used as anthelmintic compared with the rest. The frequent use of anthelmintics can favor the development of immunosuppression, drug-resistant helminths, and residue accumulation in animals (Dar et al., 2020).

The suspected resistance developed by strongyles in the study area against albendazole and ivermectin contradicts the findings of Teklemariam et al. (2016), who had no anthelmintic resistance by nematodes against those anthelmintics. The suspected anthelmintic resistance obtained in the present study might be ascribed to the factors such as prolong usage of the drugs, not adhering to the manufactures’ instructions, mass deworming using albendazole and ivermectin, substandard albendazole and ivermectin used among others (Adediran and Uwalaka, 2015).

Resistance of ivermectin was suspected for Strongyles in sheep in the study area. The resistance mechanism of macrocyclic lactones, to which ivermectin belongs, is usually due to an overexpression of genes coding for P-glycoprotein efflux pumps (Kotze & Prichard, 2016).

At two weeks post-treatments, albendazole had 100% elimination of tapeworm eggs, this is consistent with van Schalkwyk et al. (1979) and Chroust (1998). This indicates that albendazole was very effective in eliminating the eggs of tapeworms in sheep in the study area. No resistance to albendazole by tapeworm in sheep in the study area was suspected (CR = 100%). This shows that tapeworm in sheep could be well controlled using albendazole. However, levamisole had no effect on the tapeworm infections.

In conclusion, levamisole had the highest efficacy against strongyle but had no effect against tapeworm infection. Resistance to both albendazole and ivermectin by strongyles were suspected in the study area.

Acknowledgement

Our gratitude goes to the staff at the UDS Spanish Laboratory, Ghana, for their assistance during the faecal sample analysis.

Authors’ contributions

Issah Bagulo and Ibn Iddriss Abdul-Rahmancontributed the study conception, supervision and data analysis, the faecal samples were collected by Issah Bagulo and Imoro Latif, data were captured by Imoro Latif. The first draft was written by Imoro Latif. All authors reviewed and approved this manuscript

Funding

No external funding was obtained for this research

Data Availability

The dataset used or analyzed during the present study is available from the corresponding author upon reasonable request.

Ethical Approval.

The research was reviewed and approved by the University for Development Studies Institutional Review Board (UDS-IRB), Ghana.

Consent to participate

All authors have consented to participate

Consent to publication

The authors have approved to publish this work

Conflicts of interest

The authors declare no conflict of interest

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The anthelmintic efficacy of ivermectin, levamisole and albendazole against strongyle and tapeworm infections in sheep

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Version 1

Abstract

Figures

Introduction

Methodology

Study area

Study Population,design and sample size

Age categorization

Mode of application of treatment and dosage

Faecal Samples Collection

Faecal Worm Eggs and Oocyte Count

Calculating the FECRT and Cure rate

Statistical Analysis

Results

Discussion

Declarations

References

Status:

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