AN EVALUATION OF THE ANTI-COCCIDIAL POTENCY OF SOME ETHNOMEDICINAL PLANTS USED IN THE TREATMENT OF POULTRY DISEASES IN ZARIA, NIGERIA

ABSTRACT
The anti-coccidial potentials of crude extracts of some folkloric ethno medicinal plants used in the treatment of poultry diseases were evaluated andsome anti-coccidial constituents were isolated and characterized from them. A total of 19 plants belonging to 14 families associated with the treatmentof various poultry diseases were identified and tested for their anti-coccidial effects. In vitro anti-coccidial efficacies of the methanolic extracts of ten ethno-veterinary plants that are frequently used in Zaria were carried out on sporulated oocysts of Eimeria tenella; viability and population structure of the oocysts over a 96 hr period was observed. Five different dilutions of the respective crude extracts of ten ethnomedicinal plants and Amprolium® (1:25, 1:50, 1:100, 1:200, and 1:400) in distilled water were placed in separate Petri dishes labelled appropriately. 0.01ml of a suspension of freshly sporulated oocysts of E. tenella containing 500 oocysts was added to each Petri dish. The set up was kept at room temperature, semi-covered with periodic homogenization. 0.1ml suspension of each dilution was examined microscopically after 24, 48, 72 and 96 hours. Damaged oocysts and structure of parasite population were registered. Extracts of the different plants had anti-coccidial activities and there was a significant difference (P in the efficacies of the plant extracts. Azadirachta indica,Nauclea diderichiiandSolanum dasyphyllum extracts exerted the greatest percentage (363) (72.50%), (466) (93.2%) and (453) (90.6%) oocyst lethality. The observed efficacy was concentration dependent with significant difference (P< 0.05) between concentration 1:25; 1:50;1:100; 1:200 as well as 1:400 concentrations. Phytochemical screening of these plantmethanolic extracts showed that they all contain alkaloids, carbohydrates, cardiac glycosides, flavonoids, saponins, terpenoids and tannins with steroids found only in S. dasyphyllum. Acute toxicity study showed that the suspension of these extracts was not toxic when administered orally route in experimental birds at 5000mg/kg body weight. The effects of methanolic extract of S. dasyphyllum,N. diderichii and A. indica carried out in 180 coccidia-infected chickens revealed that all the infected birds in groups A1 to E1A2 to E2 as well as A3 to E3 showed clinical signs of depression, weakness, ruffled feathers and bloody diarrhoea, oocysts were detected in their faeces but absent in the non-infected birds (F1 to F3). (A1, A2 and A3 areE. tenella- infected birds and treated with Amprolium; B1, C1 and D1 areE. tenella-infected birds and treated with 1000mg/kg, 500mg/kg and 250mg/kg of S. dasyphyllum extract respectively; B2, C2 and D2E. tenella-infected birds and treated with 1000mg/kg, 500mg/kg and 250mg/kg of N. diderichii extract respectively; and B3, C3 and D3 E. tenella-infected birds and treated with 1000mg/kg, 500mg/kg and 250mg/kg of A. indica extract respectively;E1 –E3 are infected and untreated birds; F1-F3 are uninfected and untreated birds.After 7 day post treatment, no coccidia oocysts were found in the birds that had been treated with S. dasyphyllum and N. diderichii but scanty oocysts found in Amprolium and A. indica-treated groups. The birds in the infected, untreated groups were discharging oocysts of E. tenella up to the 14th day post infection. The fractions of S. dasyphyllum and N.diderichii as well as isolated substances, substance form Fractions from S. dasyphyllum and N. diderichii methanolic extracts respectively (BS1 and BS2) were tested for their in vitro anti-coccidial efficacies in comparison with Amprolium (the universal drug). Fractions 7 and 8 of the six N. diderichii fractions showed the highest anti-coccidial activity against sporulated oocysts of E. tenella. Fractions 6 and 9 of the six fractions of S. dasyphyllum were more efficient against sporulated oocysts of coccidia. BS2 was more potent against sporulated oocysts of E. tenella than BS1. The active anti-coccidial principles in the plants under study were characterized and contain betulinic and ursolic acids.Crude methanolic extracts of ten ethnoveterinary plants frequently used by poultry farmers in Zaria; fractions and substances from two (S. dasyphyllum and N. diderichii) most active plant extracts had anti-coccidial efficacies.


CHAPTER ONE

1.0 INTRODUCTION

1.1 General Introduction

The plant world is extremely diverse and many plants have a lot of known benefits. Medicinal plants are plants in which one or more of their organs contain substance(s) that can be used for therapeutic purposes or precursors for pharmaceutical synthesis (Sofowora, 1984).

The practice of herbal medicine dates back to the very earliest period of known human history. There is evidence of herbs being used in the treatment of diseases for revitalising body system in almost all ancient civilizations, the Egyptian, the Chinese and even Greek and Roman civilizations (Aftab and Sial, 1999). Herbs normally used are picorhiza, garlic, cloves, slippery elm, neem fruit and leaves, sophora flavescens, nutmeg, cinnamon, ginger, peppermint, sage, thyme, mustard and fenugreek. These plants are used as digestive stimulants, antidiarrhoic, antiseptic, anti-inflammatory, antiparasitic and appetite stimulants in human beings as well as animals (Tipu et al., 2006). A significant proportion of pharmaceutical products used currently are derived from plants (Cowan, 1999; Rankin et al., 2002). For instance more than 140 compounds have been isolated from neem tree alone (Virginie, 2010). Different plant phytochemicals such as organic acids, tannins and essential oils seem to be candidates of interest as alternative to antibiotic growth promoters (Efterpi et al., 2012). These have been shown to reduce gastro intestinal infections and increase performance in birds (Aengwanish and Suttajit, 2010; Efterpi et al., 2012).

Plants are used medicinally in different countries and are sources of many potent and powerful drugs (Lagu and Kayanja, 2010; Shivaramaiah et al., 2014).


Medicinal plants aid in healing of several diseases in poultry chickens such as diarrhoea, mange, coccidiosis, Newcastle diseases, coccidiosis, lice and tick infestation among others because the values of plant lies in some chemical substances called bioactive molecules or ingredients from plants extract (Okwu,1999; 2001; Olanipekun, 2011).

Poultry production is the most widespread in the world that forms an important component of small farmers‟ livelihoods as a tool for poverty alleviation (Dolberg, 2007). Birds are kept in a low input or low output system, with the available scavenging feed base supplemented with food scraps and grains (Sonaiya, 2007). Birds and their by-products are usually consumed by their owners, sold locally and used as gifts (FAO, 2005).

Poultry production though widespread is also faced with wide range of problems among which is infectious diseases. Poultry coccidiosis is the most studied of these infectious diseases as it has been considered a very harmful disease which affects growth and performance of birds in poultry (Bachaya et al., 2012).Coccidiosis is transmitted by direct or indirect contact with droppings of infected birds. New hosts are infected when they ingest sporulated oocysts in contaminated food or water supplies. This is referred to as faecal and or oral transmission (Guèye, 2003).

Coccidia parasite causes severe damage in chicken poultry production where chickens are reared in large numbers and high densities (Peek, 2010). Young individuals fall ill frequently, while older birds tend to be carriers (Shivaramaiah et al., 2014). Its causative agent is a single-celled protozoan belonging to genus Eimeria (Blake and Tomley, 2014; Chapman, 2014; Shivaramaiah et al., 2014).

Nine species of Eimeria are widely recognized to infect poultry chickens and each species has its own characteristics according to site of infection, immunogenicity and 2

pathogenicity. These include E. tenella, E. hagani, E. mivati, E. necatrix, E. maxima, E. brunnetti, E. mitis, E. praecox and E. acervulina (Taylor et al., 1995). The two most common species that affect poultry production worldwide are E. acervulina and E. tenella, which are the causative agents of upper intestinal and caecal coccidiosis respectively (Virginie, 2010).The main problem with Eimeria infections is that they are caused by more than one species that attack different regions of the intestine. The use of several drugs, alone or in combination, has proven to be an effective alternative in the struggle against avian coccidiosis (Quiroz-Casta˜neda and Dant´an-Gonz´alez, 2015).


Coccidiosis causes heavy economic losses to commercial poultry farmers and is thought to be one of the most expensive infectious diseases of poultry (Masood et al., 2013; Chapman, 2014). An estimated $800 million is lost world wide in the poultry industry annually as a result of morbidity and mortality associated with coccidiosis (Oluyemi and Roberts, 2000). These include costs of prophylaxis, medications, losses of productivity due to mortality, morbidity and lowered feed conversion (Williams, 1998; Chapman, 2009).The disease is clinically characterized by bloody diarrhoea, poor feed conversion ratio, low growth rate or poor weight gain. This has also been considered a contributory factor in the pathogenesis of other diseases (Bachaya et al., 2012).

In external environment, farmers usually fight this disease by applying disinfectant compounds. Commonly used disinfectants include some phenolic products such as ammonia, methyl bromide and carbon disulphide. The toxic effects of these products represent a danger to the poultry staff and health of birds and therefore their use has been restricted (Hilbrich, 1975; Williams, 2004).


1.2 Statement of the Research Problem

The anti-coccidial efficacy and phytochemical properties of some ethno medicinal plants used in the treatment of poultry diseases in Zaria have not been scientifically evaluated and therefore not fully known.



1.3 Justification

The African environment is probably the least explored in terms of available plant resources that are not yet tapped (Cowan, 1999). The use of medicinal plants all over the world started before the introduction of antibiotics and other modern drugs in the African continent (Akinyemi et al., 2005).Due to the widespread of drug resistance constraint, residual effects of drugs in meat of birds and toxic effects of disinfectants, scientists all over the world are shifting towards alternative approaches for the control of parasitic problems (Akhter et al., 2014; Hamad et al., 2014).The use of medicinal plants in the treatment of diseases have generated renewed interest in recent times, as herbal preparations are increasingly used in health care systems to treat various animal diseases especially poultry (Xiao et al., 2014).


Local farmers in Nigeria and Zaria in particular use ethno veterinary medicine (EVM) to manage chicken diseases and parasites.Ethno-veterinary practices appear to work especially to the practitioners. The plant materials are available, cheap and easy to prepare in the home at no cost and administer. Herbal medicines are also known to be broad spectrum, some have potent properties and complex bioactivity (Zhang et al., 2012)and may be the future answer to resistance of pathogen to convectional drugs (Mwale etal., 2005).Over 300,000 species of flowering plants have been recorded worldwide. So far, less than 1% of them have been explored for use against

protozoanparasitic diseases and the African continent including Nigeria have record of the least exploitation of these plants (Thangarasu et al., 2016). The scientific basis for the claim of potential has not been verified and hence the study. It is necessary, therefore, to validate the usefulness of the practice of the use of plants by local and poor farmers to treat poultry diseases towards establishing a source of developing drugs for treating coccidiosis. This research work was therefore, carried out with the following aim and objectives as stated below.



1.4 Aim

To evaluate the anti-coccidial potentials of some crude extracts of some folkloric ethno medicinal plants used in the treatment of poultry diseases and to isolate and characterize some anti-coccidial constituents.



1.5 Objectives

This study was designed to determine:

i. the folkloric plants used in the treatment of poultry diseases in Zaria;

ii. the anti-coccidial activity of the methanolic crude extracts, fractions and substances of some selected folkloric plants;

iii. the phytochemical constituents of the methanolic crude plant extracts; iv isolate some of the active anti-coccidial principles in the plants and
characterize some of the active anti-coccidial principles in the plants understudy.



1.6       Research Hypotheses

i. There are no significant folkloric plants used in the treatment of poultry diseases in Zaria.

ii. Methanolic crude plant extracts, fractions and isolated substances do not have significant anti-coccidial activity. 
iii. There is an insignificant difference in the phytochemical constituents of methanolic crude plant extracts.
iv. There are insignificant active anti-coccidial principles in the plants selected.

v. There are insignificant substances in the plants selected.

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