University of Ghana http://ugspace.ug.edu.gh UNIVERSITY OF GHANA COLLEGE OF BASIC AND APPLIED SCIENCES PET OWNERSHIP AND PARASITIC INFECTIONS IN COMPANION ANIMALS IN SELECTED COMMUNITIES IN SOUTHERN GHANA EMMANUEL OKORIE BAAH (10388298) THIS THESIS IS SUBMITTED TO THE UNIVERSITY OF GHANA, LEGON IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF MPHIL IN PARASITOLOGY DEGREE DEPARTMENT OF ANIMAL BIOLOGY AND CONSERVATION SCIENCE JULY 2019 University of Ghana http://ugspace.ug.edu.gh DECLARATION I hereby declare that the research work in this thesis is authentic and was conducted by the author. Works from other authors, where appropriate, have been duly cited. This work has not been submitted to any institution for the award of any degree. CANDIDATE EMMANUEL OKORIE BAAH SIGNATURE................................................ DATE………………………………..…….. SUPERVISORS DR. FRED ABOAGYE-ANTWI SIGNATURE……………………………… DATE………………………………............ DR. ISAAC FRIMPONG ABOAGYE SIGNATURE……………………..……...... DATE……………………….....……........... DR. DANIEL ODURO SIGNATURE................................................ DATE............................................................ i University of Ghana http://ugspace.ug.edu.gh ABSTRACT In Ghana, companion animals such as dogs are rarely given medical treatment and thus serve as reservoirs of zoonotic diseases. The close relationship of dogs and horses with humans poses risk of zoonoses. Identifying the factors that affect dog ownership and infections in dogs and horses is useful for provision of veterinary services and reducing risk of human infections. A cross sectional study was conducted to identify factors that affect dog ownership and to evaluate the factors that affect helminth infections in dogs and horses in Ghana. Using coprological, haematological and interviewing techniques, 428 dogs, 78 horses and their owners or handlers were studied. It was found out that gender, reason for keeping dogs, occupation and religion influenced dog ownership (p ˂ 0.001). The prevalence of helminth infections in dogs was 66.6%. Ancylostoma caninum (39.5%), Toxocara (40%), Dipylidium caninum (26.2%) were the zoonotic helminths identified in the study. Spirocerca lupi (3.0%) was the only helminth that was not known to be zoonotic. A chi-square analysis suggested that deworming status was significantly associated with helminth infection in dogs (p ˂ 0.001). The prevalence of Babesia spp. in dogs was 7.7%. It was found that 69.4% of dog owners never dewormed their dogs and 89.7% of respondents knew rabies as a canine zoonosis. However, none of the respondents were aware of any helminthozoonoses or protozoonoses. The prevalence of helminth infections in horses was 39.7%. The helminths identified in horses were Parascaris (32.9%), Strongyle-type eggs (26.0%) and Trichostrongylus (8.2%). Inherent factors such as age, sex and breed were not statistically associated with helminth infection in horses (p ˃ 0.05). The prevalence of Babesia spp. in horses was 11.0%. None of the stable keepers were aware of any equine-related zoonoses. It is concluded that gender, religion, occupation and reasons for keeping dog were significant factors that may influence dog ownership. Deworming status was also a significant factor ii University of Ghana http://ugspace.ug.edu.gh associated with helminth infection in dogs. The difference in proportion of dog owners based on factors such as gender, religion could be influenced by the high number of male heads in Ghanaian households and religious or socio-cultural beliefs. Variations in parasite prevalence could possibly reflect differences in pet management practices, parasite and host species, anthelminthic treatment, local climatic conditions and possibly, diagnostic techniques. Based on the limited knowledge of respondents on zoonoses, integrative approaches on creating public awareness on zoonoses, and management of companion animals is recommended to protect the health of dogs and humans. iii University of Ghana http://ugspace.ug.edu.gh DEDICATION This thesis is dedicated to my beloved mother Lydia Baah Agyakwa. iv University of Ghana http://ugspace.ug.edu.gh ACKNOWLEDGEMENTS My deepest appreciation goes to my supervisors, Dr. Fred Aboagye-Antwi, Dr. Isaac Frimpong Aboagye and Dr. Daniel Oduro of the Department of Animal Biology and Conservation Science (DABCS), University of Ghana for their efforts, inspiration and constructive criticism during the period of this study. I am very grateful to Dr. Johnson Sherry Ama Mawusi, of the Small Animals Teaching Hospital (SATH), University of Ghana for her professional advice during the design of this study. I appreciate the contributions of the laboratory technicians of DABCS during laboratory examination of samples. I am grateful to Miss Alberta Ashison (BSc. Animal Biology and Conservation Science) for her assistance in sample collection. I am thankful to Mr Ishmael Yawson and Dr Peterson of the Nsawam Veterinary Clinic for their role played in the course of this study. Most importantly, I am very thankful to the Almighty God for giving me this opportunity and my parents for their financial assistance during my study. v University of Ghana http://ugspace.ug.edu.gh TABLE OF CONTENTS DECLARATION ..................................................................................................................................... i ABSTRACT ............................................................................................................................................ ii DEDICATION ....................................................................................................................................... iv ACKNOWLEDGEMENTS ..................................................................................................................... v LIST OF FIGURES ................................................................................................................................ ix LIST OF TABLES .................................................................................................................................. x LIST OF ABBREVIATIONS ................................................................................................................. xi CHAPTER ONE ..................................................................................................................................... 1 1.0 INTRODUCTION ............................................................................................................................. 1 1.1 General Introduction..............................................................................................................1 1.2 Rationale of study .............................................................................................................................. 3 1.3 Objectives ......................................................................................................................................... 6 1.3.1 General Objective ........................................................................................................................... 6 1.3.2 Specific Objectives ......................................................................................................................... 6 CHAPTER TWO .................................................................................................................................... 7 2.0 LITERATURE REVIEW .................................................................................................................. 7 2.1 Pet ownership .................................................................................................................................... 7 2.2 Companion animals ........................................................................................................................... 9 2.3 Classification of dog populations ..................................................................................................... 10 2.4 Importance of dog and horses .......................................................................................................... 10 2.5 Impact of pet ownership on owner's health and behaviour ................................................................ 12 2.6 Parasite prevalence in free roaming pets ........................................................................................... 13 2.7 Helminthozoonoses ......................................................................................................................... 14 2.7.1 Echinococcosis ............................................................................................................................. 14 2.7.1.1 Diagnosis, treatment and control of Echinococcosis ................................................................... 15 2.7.2 Dipylidiasis .................................................................................................................................. 15 2.7.2.1 Diagnosis, treatment and control of Dypilidiasis ......................................................................... 16 2.7.3 Toxocariasis ................................................................................................................................. 17 2.7.3.1 Diagnoses, treatment and control of Toxocariasis ....................................................................... 19 vi University of Ghana http://ugspace.ug.edu.gh 2.7.4 Ancylostomiasis (Hookworm disease) ........................................................................................... 20 2.7.4.1 Diagnoses, treatment and control of Ancylostomiasis ................................................................. 22 2.8 Common haemoparasites of dogs ..................................................................................................... 22 2.8.1 Leishmania spp. (Haemoflagellate) ............................................................................................... 23 2.8.1.1 Diagnoses, treatment and control of Leishmaniasis ..................................................................... 24 2.8.2 Babesia spp. ................................................................................................................................. 24 2.8.3.1 Diagnoses, treatment and control of Babesiosis .......................................................................... 25 2.9 Gastrointestinal helminths of horses ................................................................................................. 26 2.9.1 Parascaris equorum ...................................................................................................................... 26 2.9.1.1 Diagnosis, treatment and control Parascariasis............................................................................ 27 2.9.2 Strongylus spp. ............................................................................................................................. 28 2.9.2.1 Diagnosis, treatment and control of Strongylosis ........................................................................ 29 2.10. Equine haemoparasites .................................................................................................................. 29 2.10.1 Diagnoses, treatment and control of Piroplasmosis ...................................................................... 30 CHAPTER THREE ............................................................................................................................... 32 3.0 MATERIALS AND METHODS ..................................................................................................... 32 3.1 Study area and population ................................................................................................................ 32 3.2 Study design .................................................................................................................................... 33 3.3 Estimation of sample size ................................................................................................................ 35 3.4 Sampling of dogs and horses ............................................................................................................ 35 3.5 Classification of dogs and horses ..................................................................................................... 36 3.6 Faecal collection, examination, processing and copromicroscopy ..................................................... 36 3.6.1 Rectal faecal extraction ................................................................................................................. 36 3.6.2 Examination of faecal sample ....................................................................................................... 37 3.6.3 Modified McMaster technique ...................................................................................................... 37 3.6.4 Formol ether sedimentation technique ........................................................................................... 37 3.7 Collection of blood and preparation of blood film ............................................................................ 38 3.7.1 Preparation of thin and thick film .................................................................................................. 39 3.7.2 Fixing and staining of slides .......................................................................................................... 39 3.7.3 Blood microscopy ......................................................................................................................... 39 3.8 Assessment of pet management practises and owners’ knowledge on zoonosis ................................. 40 3.9 Parasite identification ...................................................................................................................... 40 vii University of Ghana http://ugspace.ug.edu.gh 3.10 Statistical analyses ......................................................................................................................... 40 CHAPTER FOUR ................................................................................................................................. 42 4.0 RESULTS ....................................................................................................................................... 42 4.1 Prevalence of helminth infection in dogs .......................................................................................... 42 4.2 Effect of age, sex and breed of dog on helminth species prevalence .................................................. 43 4.3 Intensity of helminth infection in dogs ............................................................................................. 44 4.4 Risk factors affecting prevalence of helminth infection in dogs ........................................................ 45 4.5 Prevalence of helminth infections in horses ...................................................................................... 49 4.6 Factors affecting the prevalence of helminth infection in horses ....................................................... 50 4.7 Effect of age, sex and breed on species-specific helminth prevalence in horses ................................. 53 4.8 Intensity of equine helminth infection .............................................................................................. 53 4.9 Prevalence of canine haemoparasites................................................................................................ 54 4.10 Prevalence of equine haemoparasites ............................................................................................. 55 4.11 Socio-demographic characteristics of respondents .......................................................................... 56 4.12 Number of owned dogs .................................................................................................................. 57 4.13 Factors affecting dog ownership ..................................................................................................... 58 4.14 Dog management practices ............................................................................................................ 59 4.15 Socio-demographic characteristics of stable keepers and horses ..................................................... 63 4.15.1 Stable keepers ............................................................................................................................. 63 4.15.2 Horses ........................................................................................................................................ 63 4.16 Horse management practices .......................................................................................................... 63 4.17 Caretakers and stable keepers’ perception on equine zoonoses ....................................................... 64 CHAPTER FIVE ................................................................................................................................... 66 5.0 DISCUSSION ................................................................................................................................. 66 6.0 CONCLUSIONS, RECOMMENDATIONS AND LIMITATIONS.................................................. 82 REFERENCES...................................................................................................................................... 85 APPENDIX ......................................................................................................................................... 116 viii University of Ghana http://ugspace.ug.edu.gh LIST OF FIGURES Figure 1: Prevalence of gastrointestinal helminth infections in dogs .......................................... 42 Figure 2: Species-specific parasite prevalence among sex, breed and age of dogs. ..................... 43 Figure 3: Prevalence of gastrointestinal helminths in horses. ..................................................... 49 Figure 4: Species-specific prevalence of gastrointestinal helminths among age, sex and breed horses. ....................................................................................................................................... 50 ix University of Ghana http://ugspace.ug.edu.gh LIST OF TABLES Table 1: Chi-square and Fisher’s exact test of association of helminth infection with age, sex and breed of dog .............................................................................................................................. 44 Table 2: Intensity of helminth infection among age, sex and of dogs ......................................... 45 Table 3: A chi-square analysis of some factors associated with overall helminth prevalence in dogs .......................................................................................................................................... 46 Table 4: A chi-square test of association of some factors that affect prevalence of helminths in horses ........................................................................................................................................ 51 Table 5: Fisher’s exact test of association of age, sex and breed with species-specific helminth infection in horses. .................................................................................................................... 53 Table 6: Intensity of helminth infection among age, sex and breed of horses. ............................ 54 Table 7: Summary of socio-demographic characteristics of dog owners from the study sites. .... 57 Table 8: Summary of number of dogs owned by dog owners from the study sites ...................... 58 Table 9: A one sample chi-square analysis of differences in percentages of some variables that affect dog ownership ................................................................................................................. 59 Table 10: Knowledge, practices and attitudes of respondents with respect to dog ownership ..... 60 Table 11: Chi-square test of association of canine management practices with breed of dog ...... 62 Table 12: Summary of data on horse management practices and percentage of horses under management practices at Ayigbe town stable and Polo grounds stable. ...................................... 63 x University of Ghana http://ugspace.ug.edu.gh LIST OF ABBREVIATIONS ELISA Enzyme-linked immunosorbent assay PCR Polymerase chain reaction CAT Computed axial tomography PCR-RFLP Polymerase chain reaction- Restriction fragment length polymorphism EPG Egg per gram MAFF Ministry of Agriculture, Food and Fisheries DABCS Department of Animal Biology and Conservation Science SPSS Statistical Package for Social Scientist DNA Deoxyribonucleic acid WHO World Health Organization P Prevalence MI Mean intensity SEM Standard error of mean EDTA Ethylenediaminetetraacetic acid xi University of Ghana http://ugspace.ug.edu.gh CHAPTER ONE 1.0 INTRODUCTION 1.1 General Introduction Generally, pets are kept for companionship, recreational purposes, hunting and security (Serpell, 1986). However, the reasons for keeping pets may vary across cultural societies (Hart, 1995). The importance of pets in human life have been established by many studies (Akiyama et al., 1987; Bolin, 1987; Garrity et al., 1989). The relationship between pet ownership and human health is questionable due to difficulties in assigning a direct connection (Headey, 2003). According to Tower & Nokota (2006), pet owners are less depressed, suffer less from mental stress and have higher self-esteem compared to individuals who do not own pets. However, despite the many benefits derived from association with pets, these animals including dogs, serve as reservoir hosts for several zoonotic diseases (Damborg et al., 2016) and pose significant risk to human health and welfare (Talan et al., 1999; Weiss, 1998). Among the many pets are dogs and horses with dogs probably being the most common household pets around the world (Ugbomoiko et al., 2008). In Ghana, the number of owned household dogs is about one million, not including the many wandering dogs that stray from street to street in some communities (Veterinary Service Directorate, 2009). Pet ownership is rarely associated with companionship or recreational purposes in Ghana. Dogs in most Ghanaian households are kept for hunting in rural communities and security in most urban and semi urban households (Johnson et al., 2015). 1 University of Ghana http://ugspace.ug.edu.gh Studies have shown that companion animals such as dogs are a potential source of several zoonotic diseases, most of which can be threats to human health and well-being (MacPherson, 2005). According to McCarthy & Moore (2000), possession of a pet is a risk factor for the incidence of pet related zoonotic diseases, especially for most dog owners in Ghana (Johnson et al., 2015). In Johnson's report, dog owners who kept dogs for hunting reasons constituted the highest proportion of dog owners. Hunting dogs had the highest prevalence of zoonotic helminths such as Dipylidium, Toxocara and Ancylostoma. Helminths are the commonly encountered enteric disease-causing organisms in dogs worldwide (Ramirez-Barrios et al., 2004). Epidemiologically, helminths are also regarded as the most important parasites. In Africa, prevalence of worm infections in dogs ranges from 62.6% in Ghana (Johnson et al., 2015), 67.2% in Tanzania (Muhairwa et al., 2008), 72.5% in Nigeria (Mahmuda et al., 2012), 89.3% in Ethiopia (Mekbib et al., 2013) to 91.4% in Gabon (Davoust et al., 2008). Some of these helminths that infect dogs are known to be zoonotic and cause diseases like Hydatidosis., Toxocariasis (Overgaauw & Knapen, 2013; Sowemimo & Ayanniya, 2017), cutaneous larva migrans (Brooker et al., 2004; Khalafalla, 2011) in humans. Some studies have reported the presence of blood-borne parasites such as Piroplasma spp., Ehrlichia spp., Hepatozoon canis and Trypanosoma spp. in domestic dogs (Ahmed et al., 1994; Useh et al., 2003; Adamu et al., 2012; Nonyelu, 2013). Pregnant women, children under five years and the elderly are at risk of contracting pet related zoonotic diseases (Kourtis et al., 2014) Horses unlike dogs are less common in most Ghanaian communities and homes. They are considered as livestock pets. They are mostly used for activities such as horse riding, entertainment and agriculture (Useh et al., 2005). Horses, like many other pets are known to suffer from parasitic infections, some of which are zoonotic. Studies on the prevalence of 2 University of Ghana http://ugspace.ug.edu.gh helminths of horses have shown different prevalence under varying management systems (Boxell et al., 2004). Parasitism has been identified as the most important hinderance to the successful rearing of horses worldwide (Capewell et al., 2005). Inherent factors such as age and sex have been found to influence parasitic infections (Bucknell et al., 1995), with young horses carrying several parasites accompanied by serious manifestations. This study therefore aimed at investigating the factors that are associated with pet ownership, parasitic infections in companion animals, and risk factors that are associated with parasitic infection in companion animals. 1.2 Rationale of study Pets are an integral part of households in most countries where they share human lifestyles, bedrooms and beds (Chomel & Sun, 2011). As modern society becomes more urbanized, the presence of traditional and exotic companion animals increases in popularity (Chomel & Sun, 2011). This is evidently true in Ghana since there has been an increase in the number of exotic dog breed sellers in the Greater Accra Region over the past few years (Daily Guide, 2011). According to Daily Guide (2011), there are over 150 unconfirmed dog breeders in Accra alone. There has also been a reported increase in the number of free roaming dogs (The Mirror, 2008). Hence, one of the aims of this study was to shed light on factors influencing dog ownership. In most Ghanaian communities, the rearing of domestic animals and keeping of pets in homes are common practices. However, despite the many benefits of pets such as psychological support, improved health and companionship (Akiyama et al., 1987; Ownby et al., 2002; Parslow, 2005; Staats et al., 2008), very little attention is invested in the health of these animals, considering the 3 University of Ghana http://ugspace.ug.edu.gh fact that diseases of these animals could be zoonotic and could have serious health implications on the owners, the immediate family and the communities as a whole (Amissah-Reynolds et al., 2016). Several studies attest to the fact that some infections of companion animals could be transmitted to humans and vice versa (Katagiri & Oliveira-Sequeira, 2007; Khante et al., 2009; Damborg et al., 2016), particularly in some rural areas where dogs are kept together with other livestocks (Chrieki, 2002). Between 2002 and 2010, numerous outbreaks of reported zoonotic diseases were identified and found to be associated with household pets (Smith & Whitfield, 2012). In developing countries such as Ghana, the greater number pet owners live under poor conditions and pet management as well as disease control strategies are non-existent. Zoonotic infections should therefore be of great concern since majority of emerging infectious diseases are zoonotic (Andersen, 1997; Oliveira-Sequeira et al., 2002; Chomel et al., 2007). A human case of Toxocara infection with prevalence of 53.5% was reported in children in the Central Region of Ghana (Kyei et al., 2015). The continuous growth of the dog breeding industry in Ghana has necessitated the need for veterinary interventions to mitigate the impact of zoonoses on public health. Epidemiological reports suggest that free roaming dogs have highest prevalence of parasites (Aberes et al., 2013; Bhattacharjee & Sarmah, 2013). Prevalence of helminth infections is high in dogs of owners who rear them for hunting purposes (86.4%) and security purposes (59.2%) (Johnson et al., 2015). A study by Johnson et al., 2015 showed that dog owners who bought their dogs from street dog sellers in Ghana had dogs that were significantly infected with helminths. An understanding of the demographics and factors that are associated with dog ownership may be of public health and veterinary importance (Robinson & Pugh, 2002), social psychology 4 University of Ghana http://ugspace.ug.edu.gh (Akiyama et al., 1987; Bolin, 1987; Garrity et al., 1989; Raina et al., 1999), or marketing of veterinary services or products. Generally, infections in pets are mostly due to the fact that pets are seldomly treated for parasitic diseases and laws on pet management and control are non-existent (Ugbomoiko et al., 2008) or poorly implemented. As such this study also sought to ascertain management practices and their effect on parasite prevalence. The prevalence of helminth parasites varies substantially from one region to another based on the helminth species involved, the host species, and climatic conditions and even management practices (Robertson et al., 2000; Oliveira-Sequeira et al., 2002). It is therefore imperative for timely investigations of the prevalence of these parasites to be conducted within a given area so that effective parasite control strategies in pets could be achieved. Horses, including most domestic animals are vulnerable to parasites and may be infected with different species (Wannas & Dawood, 2012). Due to the economic importance of enteric parasites, huge amounts of money are invested in the control of these parasites worldwide (Bliss, 2018). Yet, enteric parasites are still a constant menace affecting the health and well-being of horses and other domestic pets in different parts of the world (Mbafor et al., 2012). Horse riders, horse keepers and stable cleaners or caretakers are the population at risk of contracting horse zoonoses. Very little data exists on pet ownership in Ghana. This study therefore addresses factors that are associated with dog ownership in Ghana, management practices and their association with helminth prevalence. 5 University of Ghana http://ugspace.ug.edu.gh 1.3 Objectives 1.3.1 General Objective 1. To investigate the factors that affect dog ownership and, parasitic infections in dogs and horses. 1.3.2 Specific Objectives 1. To determine the prevalence of helminths in dogs and horses. 2. To determine the prevalence of haemoparasites in dogs and horses. 3. To determine the association between pet keeping/management practices and helminth prevalence in dogs and horses. 4. To assess the knowledge, attitudes and practices of dog and horse owners in their management. 6 University of Ghana http://ugspace.ug.edu.gh CHAPTER TWO 2.0 LITERATURE REVIEW 2.1 Pet ownership Humans have been keeping pets for many years (Serpell, 1986). Pet ownership is a common practice in many households in many countries (AVMA, 1993; Butler & Bingham, 2000; Murray et al., 2010). Although companion animals (pets) may be defined in many ways (Eddy, 2003), cats and dogs are conventional paradigms of pets. In Canada, it was estimated that 56% of homes owned a dog or cat (Perrin, 2009). In the USA, about 56% of houses constituting 53 million have companion animals (APPMA, 1994). In South Africa, it has been reported that 37% of homes that keep companion animals kept dogs and cats, horses and birds (Odendal, 1994). An estimated 2% of the households in USA own an average of 2.54 horses (AVMA, 1993). Pet ownership in European homes include Germany (37%), Ireland (70%), Britain (55%) and France (63%) (Reader's Digest Association Inc, 1991). Pet ownership is associated with spending of time and financial resources. An estimated 40.8 billion dollars is spent yearly on pet industry in United States (APPMA, 2007). Pet ownership is also generally related to demographics and there are various definitions of pet and pet ownership (Salman et al., 1998; Hart, 2003), yet there is little data regarding the reasons for having pets. In Ghana, pets are kept for several reasons. Some Ghanaians keep pets such as dogs for hunting. Others keep dogs for security reasons and few keep dogs for recreational and companionship purposes. A study conducted by Johnson et al., 2015 reported 27.1% and 56.8% of dogs were kept for hunting and security respectively whilst 8.4% and 7.6% were kept for companionship and breeding purposes respectively in the Greater Accra region of Ghana. 7 University of Ghana http://ugspace.ug.edu.gh Many studies have been conducted on the factors associated with dog ownership some in developed countries (Franti & Kraus, 1974; Westgarth et al., 2007). However, only a few studies have been conducted in developing countries (Knobel et al., 2008). In Tanzania, pet ownership is largely observed among better educated, large and wealthier households (Knobel et al., 2008). In this same report, it was observed that households with Muslim occupants were less likely to own dogs than households with Christian occupants. It was also reported that households that had male heads were more likely to own a dog. Several reports in Sub-Saharan Africa indicate that the number of unowned dogs is low (Knobel et al., 2008). The number of the dogs owned varies significantly worldwide. In Africa, many dogs are not restricted and hence, stray freely (Beck, 2000). In Chad, 10.9% of the total dog population is not owned and are considered stray dogs. In Sri Lanka, 19% of the dog population is not owned (Kayali et al., 2003). In Tunisia, about 5-15% of the dog population was considered “stray”. The breeding of companion animals such as exotic dog breeds has become very common in Ghana. Unconfirmed reports indicate that there are over 150 dog breeders in Accra alone. The dog trade industry in Ghana is a diverse mix of small scale and large-scale systems which comprises a range of street side pet sellers to well organized breeder houses in large cities. The interest and demand of many Ghanaians to pet exotic dog breeds has made the dog trade industry lucrative. This has attracted many others to join the dog trade industry. Dog breeders in Ghana sell puppies at price of 500 to 2,000 GHC depending on the breed of dog. However, local dog breeds are very cheap, hence are common in many Ghanaian homes, resulting in growing population of owned and stray dogs. 8 University of Ghana http://ugspace.ug.edu.gh Horses are not common household pets in Ghana. In Ghana, horses are kept for games in polo clubs, rides on beaches and parade in the military and police force. In some communities, especially the Northern part of Ghana, horses are a vital part of festivals where chiefs ride on beautifully adorned horses. According to a stable keeper at the Accra Polo grounds, horses are owned by several stakeholders who enrol their horses in polo games for profit and sometimes offer them for sale. 2.2 Companion animals A companion animal or pet is a tamed animal that is kept for pleasure rather than utility (Eddy, 2003). This description of a pet focuses on an animal that is kept for pleasure and companionship, while specifically excluding utilitarian contribution to labour, health and perhaps security. Large animals such as horses, camels, donkeys and elephants that are used for transportation and for carrying loads may fill a role as companions and may be strongly attached to their human handlers. Fishes, gerbils, birds, rabbits or hamsters, frogs, lizards, snakes, guinea pigs and turtles have been reported in some homes as pets (Perrin, 2009). A survey conducted in 1992 on companion animals in South Africa reported that there were about 393 million dogs, 1 million horses, 98 million birds, 97 million cats and 22 million other companion animals (Odendal, 1994). Dogs are the commonest household companion animals with horses being the least common (Odendal, 1994). In Ghana, little information exists on the demographics of companion animals except that an annual census is taken by the Veterinary Service Directorate of Ghana. However, this census only reflects the number of reported cases at the various veterinary clinics, and not all pet owners are veterinary clients. According to the Veterinary Service Directorate (2009), Ghana has a 9 University of Ghana http://ugspace.ug.edu.gh population of about one million owned dogs. No reliable information currently exists on the demographics of other companion animals in Ghana since a comprehensive study on the demographics and census of companion animals has not been conducted. Pfukenyi et al. (2010) reported that 82% of respondents owned dog in Zimbabwe in a survey of pet ownership. The global dog population is estimated to be more than 500 million (Hsu et al., 2003). 2.3 Classification of dog populations The different relationships between cultures and the dogs associated with them has varied the structures of canine population. The classifications of dog population are based on the degree of dependence and supervision by humans. According to Matter & Daniels (2000), three types of dog populations have been classified: dogs that do not roam without human supervision; dogs that stray or roam without human supervision; and dogs that are not owned and freely roam. WHO (1990) classifies dog population as: restricted dogs that are supervised and are dependent on humans for resources; family dogs that semi-restricted, stray partly and are dependent on humans for food; neighbouring dogs that are semi-restricted, roam freely and are semi-dependent on humans for food; feral dogs that are unrestricted and are not dependent on humans for resources. 2.4 Importance of dog and horses Pets such as dogs are kept in households for security and companionship (Serpell, 1986). In Ghana, Johnson et al. (2015) reported that dogs were kept in homes for hunting, security and companionship reasons. In Zimbabwe and Kenya, dogs are appreciated for the security they offer both humans and livestock against invaders (Butler & Bingham, 2000). According to Serpell (1986), Marvin (1998), and Podberscek (2009), dogs are a delicacy in some cultures. In some 10 University of Ghana http://ugspace.ug.edu.gh parts of West Africa and Southeast Asia, dogs are important source of protein in diet. In Nigeria, dogs serve as meat in some local communities (Opara et al., 2004; Ayinmode et al., 2016). However, in a report by Gadegbeku et al. (2013), 64% of respondents in Ashongman-Accra in Ghana considered dog meat as a food taboo for religious, ethical, superstitious and compassionate reasons. In the Islamic religion however, dogs are considered to be spiritually impure (Beck, 2000). The relationship between dogs (stray or communal) and humans is not always postive. In South Africa, dogs cause problems such road accidents, barking, biting children and livestock, and faecal contamination (Beck, 2000). In some developed counties, dogs are important in forensic investigation, being able to locate people or illegal drugs (Stejskal, 2012; Alexander et al., 2015). Sniffer dogs have been used by law enforcement agencies in legal investigations for detection of drugs, explosives, arson, cadaver, search and rescue, tracking, training and fieldwork for decades (Stejskal, 2012). Horses are utilized through activities such as horse riding and police service (Patterson, 2002), transportation (Greene, 2008), agriculture (Thompson, 1976) and historically in warfare (Stanek et al., 2004). Curative horse-riding programs have shown positive effects on the well-being of people with disabilities (Yorke et al., 2008). Many products such as meat, milk, hide, hair, bones are derived from horses (MaCGregor, 1985; Chong-Eon et al., 2007). Drugs such as Premarin, a menopausal oestrogen replacement therapy is extracted from the urine of pregnant horses (Tempelman-Kluit, 1999). Horse meat is eaten in many parts of the world (Ambriz, 2004; Chong- Eon et al., 2007). The bone of horse is used to make implements (MaCGregor, 1985), the hooves are used for making animal glue (Bartlett, 1994) and skin as the leather for making jackets and baseball gloves (Ockerman & Hansen, 2000). The tail hair of horses can be used for making string musical instruments such as violin and viola (McCutcheon, 2000; Matsutani, 2002). 11 University of Ghana http://ugspace.ug.edu.gh 2.5 Impact of pet ownership on owner's health and behaviour Evidence suggests that pet ownership is beneficial to human health and behaviour (Serpell, 1991). Friedman et al. (1980) reported significant statistical association between pet ownership and survival of patients. However, the approaches used in Friedman's work have been criticised (Wright & Moore, 1982), and the results have not been reproduced (Serpell, 1991). Other reports indicate that if a person is strongly attached to animal companion, it may help lessen the impact of negative life events, such as grief, and have positive effect on anxiety and depression (Akiyama et al., 1987; Bolin, 1987; Garrity et al., 1989). A decline in risk of asthma, allergic rhinitis and cardiovascular diseases has been reported in people exposed to pets (Anderson et al., 1992; Nafsted et al., 2001; Ownby et al., 2002). Reduced symptoms and risk of depression have been observed in the aged with strong pet attachments (Garrity et al., 1989). Siegel (1990) also reported that people who own pets have less stressful life. However, other studies have failed to detect a relationship between pet ownership and improved health status (Ory & Goldberg, 1983; Robb & Stegman, 1983). Parslow et al. (2005) reported depression and poorer physical health among pet owners. Researchers have also demonstrated that pet ownership among the elderly provides no health benefits (Tucker et al., 1995; Simons et al., 2000) or emotional distress (Stallones et al., 1991). Boredom and loneliness have also been reported among pet owners (Wells & Rodi, 2000). Other researchers also failed to prove that pet ownership is linked with a reduced risk of cardiovascular diseases (Parslow & Jorm, 2003) or any psychological or physical health improvement of the elderly (Parslow et al., 2005). 12 University of Ghana http://ugspace.ug.edu.gh 2.6 Parasite prevalence in free roaming pets Free-roaming cat and dog populations have been identified as risk to public health (Traub et al., 2005; Garde et al., 2013; Gerhold & Jessup, 2013). Free-roaming dogs and cats lack the veterinary care to control zoonotic diseases and hence, pose a potential health hazard to humans and other animals (Gerhold & Jessup, 2013; Garde et al., 2013). Responsible pet owners are able to significantly control pet related zoonotic diseases through proper routine veterinary care and vaccinations (Gerhold & Jessup, 2013). However, in a study conducted by Amissah-Reynolds et al., 2016 in Ghana, 46% of 100 respondents never dewormed their dogs. This suggests that only a few pet owners exercise dog veterinary care. Amissah-Reynolds and colleagues, however did not report the prevalence of helminth infection in dogs that were not dewormed. Johnson et al. (2015) in their report on management practices of dogs reported helminth prevalence in dogs that received anthelminthic treatment. The result was however statistically insignificant. Effective dog management is seldomly achieved because fewer resources are assigned towards dog health and control (Dalla et al., 2010). A study of helminths in stray dogs reported a prevalence of 2%-43% in in Iran (Emamapour et al., 2013), 84.78% in stray dogs in Ethiopia (Aberes et al., 2013) and 66.7% in Ghana (Amissah- Reynolds et al., 2016). Johnson et al. (2012) reported significant helminth infection in free range dogs as compared with dogs restricted within walled houses. Dogs and cats act as reservoirs of various zoonotic infections (Labarthe et al., 2004; Traub et al., 2005). These animals live freely in the community and contaminate the environment with helminths and other protozoans discharged in their faeces (Traub et al., 2005; Zibaei et al., 2007; Arbabi et al., 2009). 13 University of Ghana http://ugspace.ug.edu.gh 2.7 Helminthozoonoses Gastrointestinal parasites in pets are common in developing countries (Perera et al., 2013). Enteric parasites are the main cause of diseases in dogs (Hendrix & Blagburn, 1983). In Ghana and other countries, gastrointestinal parasites cause significant public health problems (Perera et al., 2013; Amissah-Reynolds et al., 2016). The association of pet owners with their pets presents a great risk of zoonoses (Aberes et al., 2013; Dhaliwal & Prayag, 2013; Amissah-Reynolds et al., 2016), despite their beneficial importance such as mental health and even physical well-being (Akiyama et al., 1987). Zoonotic diseases are cosmopolitan and are spread through contact of humans with companion animals or livestock. 2.7.1 Echinococcosis Echinococcosis is a zoonotic disease that is caused consuming the hydatid cyst of Echinococcus granulosus and Echinococcus multicularis (Schmidt et al., 2009). In many developing countries, dogs serve as means of disposing offal and hence, they may get infected when they consume the viscera of an infected animal (Schmidt et al., 2009). In the life cycle of the worm, ingested cysts attach to the small intestines and develop. Eggs are produced and are passed in the dog’s faeces where they contaminate the environment and are ingested by other animals including humans. Human infection results when eggs of Echinococcus spp. are unintentionally ingested usually as a result of caressing dogs. In human infection, cysts develop commonly in the liver, lungs or brain. Some tribes of Kenya have the highest rate of Echinococcosis since they relish roasted dog intestines (Nelson & Rausch, 1963). In Lebanon, leather tanners use dog faeces to tan leather and risk contamination (Schwabe & Daoud, 1961). 14 University of Ghana http://ugspace.ug.edu.gh 2.7.1.1 Diagnosis, treatment and control of Echinococcosis Hydatids can be seen using X-radiography, ultrasonography or CAT scans. Immunodiagnostic methods are less sensitive than imagery (Babba et al., 1994). Molecular diagnostic techniques such as ELISAs and PCR test for Echinococcus spp. are considerably effective (Craig & Larrieu, 2006; Conraths & Deplazes, 2015). Surgery is the only method of treatment when the cyst is found in an unrestricted location (Moro et al., 2000). Albendazole is the drug of choice for treatment of Echinococcosis. Elimination can be achieved by refusing dogs access to offal, culling stray and free roaming dogs, and by education of the general public by veterinary services (McManus & Smyth, 1986; Gemmell et al., 1987). 2.7.2 Dipylidiasis Dipylidiasis is a zoonotic disease caused by Dipylidium caninum, a cosmopolitan tapeworm found in dogs and cats. The oval gravid segment of the worm is elongate and is shaped somewhat like a cucumber seed. It contains egg capsules or packets, each containing as many as 20 eggs (Griffiths, 1978). The life cycle involves an intermediate host that may be the dog or cat flea (Chen, 1934) or louse and human flea (Bartsocas et al., 1966; Craig & Ito, 2007). Gravid tapeworm segments are released in host's faeces. Eggs are ingested by the adult louse or flea larvae. Thiodectes canis and Pulex irritans also act as intermediate hosts (Neira et al., 2008; Ramana et al., 2011). The active oncosphere develops to the infective cysticercoid stage in the arthropod host (Robertson & Thompson, 2002). The definitive host becomes infected by consuming the infected arthropod. Chronic enteritis is common in heavy infections. Nervous disturbance and tension are attributed to the tapeworm infections and may be associated with such 15 University of Ghana http://ugspace.ug.edu.gh clinical signs as abdominal discomfort, vomiting, convulsions and severe epileptic fits (Bradley, 1971). Gravid segments around the anus may cause anal irritation (Dantas-Torres, 2008). A human case of zoonoses caused by Dipylidium caninum has been reported in China (Peng et al., 2017) and in the United States (Turner, 1962; Hunter, 1962). Infants and young children are at risk of contracting Dipylidiasis due to their playing habits with pets (Bartsocas et al., 1966; Casasbuenas, 2005; Peng et al., 2017). A report from Chile indicates that children between the ages of 2 months to 4 years old are the population at risk of contracting Dipylidiasis (Neira et al., 2008). Accidental infection occurs when the flea vector is swallowed or when people kiss or are licked by their pet (Schmidt et al., 2009). Dipylidiasis has a worldwide distribution (García- Agudo et al., 2014). Cases have been reported in children and adults in Europe, Japan, China, United States, India, Sudan and Latin America (Neafie & Marty, 1993; Brandstetter & Auer, 1994; Vargas et al., 2000; Molina et al., 2003; Casasbuenas, 2005; Tsumura et al., 2007; Samkari et al., 2008; Zhao et al., 2008; Li et al., 2010; Cabello et al., 2011). Another case of human dipylidiasis has also been reported in girl from Spain (García-Agudo et al., 2014). In the report it was found out that the girl's family owned a dog with no flea collar and poor veterinary care. Another case of dipylidiasis was reported in Sinaloa, Mexico (Cabello et al., 2011). 2.7.2.1 Diagnosis, treatment and control of Dypilidiasis Diagnosis of D. caninum is done by observing rice grain-like proglottids and egg packets in the parasite (Molina et al., 2003; Neira et al., 2008). Identification of tapeworm segments crawling on stool or on the perianal hairs of dogs is indicative of infection. Tapeworm eggs can be found on faecal examination if segments have been crushed on passage through the anal sphincter or mechanically broken as the faeces are stirred (Griffiths, 1978). Children expel proglottids in 16 University of Ghana http://ugspace.ug.edu.gh faeces and they may be found on the anus or diapers (Reis et al., 1992; Bowmann, 1995). García- Agudo et al. (2014) reported that the incidence of human dipylidiasis is unknown, due to misdiagnosis. Treatment of dipylidiasis has been done with various drugs such as paromomycin, and niclosamide (Jones , 1979) and praziquantel (Wijesundera, 1989; García-Agudo et al., 2014). 2.7.3 Toxocariasis Toxocariasis is a globally distributed zoonosis caused by Toxocara canis and Toxocara cati. T. canis infects dogs and is rare in cats (Anderson, 2000). Birds and livestock (e.g. sheep, pigs and poultry) serve as paratenic hosts (Taira et al., 2004). Adult worms inhabit small intestines and somatic tissues of their host. Adult T. canis may be 18 cm long, the sub globular egg, about 85µm in diameter, is finely pitted (Anderson, 2000). Surveys have shown that virtually all puppies born are infected with T. canis (Barriga, 1988). A survey in Nigeria reported a prevalence of 30.5% T. canis infection with 39.5% of the dogs between the ages of 0-6 months (Sowemimo & Ayanniya, 2017). In the United States, more than 30% of dogs less than 6 months were found to be shedding T. canis eggs (Blagburn, 2001). Infection is less common in adult dogs but it still occurs (Blagburn, 2001). In the cycle, adult parasite sheds eggs in the faeces of the host. The eggs develop and become infective to the third stage larvae. The larvae invade the host through ingestion of the third stage larvae. The larvae hatch in the digestive system and migrate towards the liver and then move to other organs (Mok, 1968; Woodruff, 1970). Migration in the lungs results in larvae breaking across the alveoli, travelling up the respiratory tract, and then being swallowed to begin egg production in the small intestines (Schmidt et al., 2009; Anderson, 2000). 17 University of Ghana http://ugspace.ug.edu.gh Foetal puppies acquire T. canis in utero when larvae from the dam's somatic tissue are activated late in pregnancy to migrate across the placenta (Burke & Robertson, 1985; Urquhart et al., 1996). The dam appears resistant to parasite and development of larvae to the adult stage is suppressed (Scothorn et al., 1965). Puppies are then born with developing ascarids (Urquhart et al., 1996). A small number of larvae may also be transmitted to nursing puppies through transmammary route in the mother's milk (Burke & Robertson, 1985; Gillespie, 1988). Transplacental transmission results in more infections than the transmammary transmission (Burke and Roberson, 1985). Infected dogs may show signs of mild to moderate cough accompanied by a mucopurulent discharge (Urquhart et al., 1996). Severe pulmonary oedema, haemorrhage and even death may result just a few days after birth (Urquhart et al., 1996). Adult ascarids in puppies can induce mucoid enteritis, which may be associated with mild diarrhoea (Griffiths, 1978; Urquhart et al., 1996), anorexia, vomiting, ill thrift, stunting and abdominal discomfort. A characteristic potbelly appearance may develop in heavily infected dogs (Griffiths, 1978). Intussusception and intestinal obstruction due to numerous worms in the small intestine of young puppies have been reported (Lyndell & Michael, 1992; Griffiths, 1978). Poor hair coat and reduced growth have been reported in infected dogs (Schneider et al., 2011). Human Toxocariasis continues to plague humans despite the anthelminthic treatment and regimens given to dogs and cats in some parts of the world (Overgaauw & Knapen, 2013). Prevalence of T. canis and T. cati vary from 3.5- 34% for T. canis in dogs and from 8-76% for T cati in cats (Fok et al., 2001; Habluetzel et al., 2003; Le Nobel et al., 2004; Dubná et al., 2007; Martínez-Moreno et al., 2007; Lee et al., 2010). Human infections usually occur when food products contaminated with the eggs Toxocara spp. are consumed (Glickman & Schofer, 1987; 18 University of Ghana http://ugspace.ug.edu.gh Nagakura et al., 1989; Stürchler et al., 1990; Baixench et al., 1992; Salem & Schantz, 1992; Taira et al., 2004). Dog contact may be an important source of transmission of Toxocara eggs to pet owners (Wolfe & Wright, 2003; Aydenizöz-Özkayhan et al., 2008; Roddie et al., 2008). Cases of human Toxocariasis have been reported in Africa (Lötsch et al., 2017), America (Lee et al., 2014; Woodhall et al., 2014), Nigeria (Okewole, 2016), Kenya and Tanzania (Wiseman & Woodruff, 1971) and Ghana (Kyei et al., 2015). 2.7.3.1 Diagnoses, treatment and control of Toxocariasis Diagnosis is established on clinical manifestations, laboratory examinations, molecular and sero- diagnostic methods (Overgaauw & Knapen, 2013). Asymptomatic infections also commonly occur (Pawlowski, 2001). Laboratory diagnosis of T. canis infection involves the confirmation of characteristic globular eggs with thick, pitted shells. Whole worms in vomitus or faeces may be identified. ELISAs using specific secretory-excretory antigens and liver biopsies are also employed in clinical diagnosis (Schmidt et al., 2009). Computer tomography, sonography and MRI are used in diagnosis of Toxocara infection (Magnaval & Glickman, 2006; Lim, 2008). Adult worms in dogs may be satisfactorily removed with piperazine and organic phosphorus compounds (Griffiths, 1978). Mebendazole as a drug for treatment is effective in human infections (Magnaval, 1995). Febendazole has also been used in the treatment of Toxocariasis in dogs (Overgaauw & Knapen, 2013). To reduce vertical transmission of T. canis from dam to puppies, topical selamectin is administered to pregnant bitches and lactating dams at 6 mg/kg (Payne-Johnson et al., 2000). Febendazole is also administered orally at 50 mg/kg daily (Burke & 19 University of Ghana http://ugspace.ug.edu.gh Roberson, 1983). Ivermectin is administered orally at 300 µg/kg body weight (Payne & Ridley, 1999). Control programmes consist of routine deworming of household pets, and proper disposal of faeces (Schmidt et al., 2009). Preventing defecation of pets in public, hygiene, and public education are some control actions against Toxocara infection (Glickman & Shofer, 1987) 2.7.4 Ancylostomiasis (Hookworm disease) Ancylostoma caninum is common cosmopolitan parasitic bloodsucking hookworm that causes a disease known as Ancylostomiasis in dogs (Meyers et al., 1976). The definitive host of this nematode parasite include dogs, foxes and humans for A. caninum; dogs, cats and foxes for A. braziliense (Griffiths, 1978). Eggs are usually laid in the 8-cell stage and are 60 × 40µm (Griffiths, 1978). A. caninum has been reported throughout the United States and Africa (Croese et al., 2013; Aberes et al., 2013; Johnson et al., 2015 Amissah-Reynolds et al., 2016). In the life cycle, the host releases eggs in the faeces after entry of parasite into the host (Yoshida et al., 1974). The third-stage larvae A. caninum can be transmitted orally (Foster & Cross, 1934). Dogs can also become infected with A. caninum upon skin penetration (Loos, 1898). The larvae migrate in the bloodstream to the lungs, coughed up and are swallowed to develop into adults in the small intestine and lay eggs (Schmidt et al., 2009). Larvae that migrate to the lungs remain in circulations and develop into quiescent forms in the muscles of dogs (Lee et al., 1975; Little et al., 1983). If the dog is a female, the quiescent larvae will become active late in pregnancy, move to the mammary glands and be transmitted to the suckling puppies (Stone & Girdeau, 1966). Such transmammary infections can result in large numbers of larvae being passed directly into neonatal 20 University of Ghana http://ugspace.ug.edu.gh puppies. The quiescent larvae may also be reactivated in adult dogs in the absence of pregnancy or lactation, migrating to the small intestine to establish a patent infection even in the absence of exposure to infectious stages in a contaminated environment (Urquhart et al., 1996). Transplacental transmission occurs in pregnant bitches which are infected cutaneously or orally and worms do not mature in the puppies until they are born (Clapham, 1962; Foster, 1932; Foster, 1935). Clinical manifestation in puppies and young dogs presents acute anaemia because of blood loss induced by the worms in the intestines (Urquhart et al., 1996). This anaemia is severe and may be fatal in neonatal puppies that acquire large numbers of larvae by transmammary transmission (Urquhart et al., 1996). In puppies as young as 10 days old, bloody diarrhoea, anaemia, hypoproteinaemia and death may ensue. In older dogs, non-regenerative iron deficiency anaemia may result. In a contaminated environment, larvae can penetrate the skin and may cause dermatitis (Schantz, 1994). Human infection due to zoonotic ancylostomiasis is rare but have been reported in many parts of the world (Anten & Zuidema, 1964; Velasquez & Cabrera, 1968; Yoshida et al., 1968; Areekul et al., 1970; Chowdhury & Schad 1972; Croese et al., 1994a; Croese et al., 1994b; Khoshoo et al., 1995; Bahgat et al., 1999; Traub et al., 2008; Jiraanankul et al., 2011; Sato et al., 2011; Dhaliwal & Prayag, 2013; Inpankaew et al., 2014). Studies in Australia show that intestinal infection with Ancylostoma caninum is a major cause of human eosinophilic enteritis (Prociv & Croese, 1996). The infection is accompanied by abdominal pain and eosinophilia (Prociv & Croese, 1996). Studies in Asia, have demonstrated that hookworm of dogs and cats is the second most common hookworm species infecting humans (Traub, 2013). 21 University of Ghana http://ugspace.ug.edu.gh 2.7.4.1 Diagnoses, treatment and control of Ancylostomiasis Diagnosis can be done by finding parasite ova in standard faecal flotation using saturated salt or sodium nitrate solution. Concentration techniques can be used to estimate faecal egg counts (Cross, 2000). Finding hookworm ova or worms in faeces is the only conclusive diagnosis of hookworm disease (Schmidt et al., 2009). Speciation hookworm can be accomplished by culturing larvae from faeces (Schmidt et al., 2009). Diagnosis can also be done with the use of molecular techniques such ELISA and western blot techniques (Sato et al., 2010). Speciation with PCR-RFLP has also yielded promising results (Hawdon, 1996). To reduce the burden of transmammary transmission of A caninum from dam to puppies, anthelminthics such as 10% imidacloprid plus 2.5% moxidectin topical solution is administered at day 56 of gestation (Kramer et al., 2006). Febendazole is administered at 50 mg/kg daily in dogs (Burke & Roberson, 1983). Ivermectin is administered intramuscularly at 300 µg/kg post conception (Stoye, Meyer, & Schneider, 1989). An irradiated larva vaccine that was extremely effective against Ancylostoma caninum in dogs was discontinued mainly because of its lack of acceptance by the American veterinary profession (Miller, 1978; Urquhart, 1980). Sanitory disposal of faeces is vital to the control of hookworm disease (Schmidt et al., 2009). 2.8 Common haemoparasites of dogs Canine haemoprotozoan parasites have been reported in dogs in Africa (Opara et al., 2016; Nonyelu, 2013; Anise et al., 2018) and other parts of the world (Bhattacharjee & Sarmah, 2013; Priyowidodo et al., 2018). Haemoprotozoans are unicellular organisms under the sub-kingdom Protozoa (Molyneux & Ashford, 1983). Transmission to their host is usually by the bite of arthropod vectors such as ticks (Birkenheuer et al., 1999) or by ingestion of an arthropod 22 University of Ghana http://ugspace.ug.edu.gh intermediate host. Haemoprotozoan infections are common in dogs in tropical countries (Radostis et al., 2000). According to Shirvastava (2014) and Tsegay (2016) canine Babesia spp., Leishmania and Trypanosoma are common haemoprotozoan parasites of dogs that result in considerable financial loss arising from treatment and high mortality rate. 2.8.1 Leishmania spp. (Haemoflagellate) Leishmania spp. are a haemoprotozoan parasites that cause Leishmaniasis. The host of the parasite includes human beings, dogs, cats, horses and sheep, with dogs and rodents principally serving as reservoir hosts for spread of zoonotic visceral leishmaniasis (Canavate et al., 2004; Ahuja et al., 2006; Chappuis et al., 2007; Schmidt et al., 2009). The parasite is transmitted by a phlebotomine sand fly (Pearson & Sousa, 1996; Chappuis et al., 2007). The parasite is found in the reticulo-endothelial cells of capillaries, spleen and other organs of the host. Parasites may also be seen in monocytes, polymorphonuclear leucocytes, and macrophages (Schmidt et al., 2009). L. donovani affects visceral areas. Parasites are small intracellular, non-flagellated oval or circular bodies about 2 µm wide and 4 µm long. The parasite occurs as a leptomonad in the insect host (Griffiths, 1978). Leishmaniasis could occur due to more than twenty Leishmania species. 90% of visceral leishmaniasis cases have been reported from five countries: India, Bangladesh, Nepal, Sudan and Brazil (NCID, 2006). Visceral leishmaniasis occurs due to L. donovani complex in Indian, East Africa and L. infantum in Africa, Europe and Latin America (Chappuis et al., 2007; Lukes, 2007). L. infantum causes zoonotic fatal visceral leishmaniasis (Dhaliwal & Prayag, 2013). Visceral leishmaniasis has been reported in a girl in India (Dey et al., 2007). A case of advanced kala-azar has been reported in a boy of about 6 years old in Sudan (Hoogstraal & 23 University of Ghana http://ugspace.ug.edu.gh Heyneman, 1969). In dogs, infection is accompanied by lesions, alopecia, lymphadenopathy, inflammation, fever, anaemia and splenomegaly (Dhaliwal & Prayag, 2013). 2.8.1.1 Diagnoses, treatment and control of Leishmaniasis Diagnosis of Leishmania spp. is based on identifying Leishman-Donovan bodies in tissues or secretions (Schmidt et al., 2009). Molecular and immunodiagnostic tests are sensitive and can detect Leishmania parasites (Schmidt et al., 2009). Molecular and biochemical methods that hybridizes mitochondrial kinetoplast DNA have yielded promising results in the identification of Leishmania (Schoone et al., 1991; De Brujin & Barker, 1992; Massamba & Mutinga, 1992). Treatment is accompanied by inoculation of antimony compounds. The drug of choice used is miltefosine (Sundar et al., 2002). Antimonials such as meglumine antimoniate (Glucantime) and allupurinol can be administered to cure Leishmaniasis. The most effective means of preventing Leishmania infection is through the utilization of topical insecticides, collars and spot-on formulations of pyrethroids on dogs. Vaccines can be used in combinations with repellants and ectoparasticides. Dogs can be housed indoors, ideally in a fine mesh netted environment to decrease sand fly bites. 2.8.2 Babesia spp. Babesia spp. are haemoprotozoan parasites that reside in the red blood cells of mammalian hosts, Babesia microti (Kjemtrup & Conrad, 2000), Babesia divergens (Telford & Spielman, 1997; Beattie et al., 2002), Babesia duncani (Bloch et al., 2012) and Babesia venatorum (Jiang et al., 2015) cause zoonotic babesiosis. Babesia divergens was first reported in human in Croatia in 24 University of Ghana http://ugspace.ug.edu.gh 1956 and epidemiological surveys have shown that B. divergens may extend beyond Europe into North Africa (Zintl et al., 2003). Canine babesiosis is caused by Babesia vogeli and Babesia gibsoni. Most human cases of Babesiosis are caused by Babesia divergens (Genchi, 2007). Canine Babesia parasites are distributed throughout tropical and sub-tropical regions. Peracute babesiosis is accompanied by collapse due to hypotensive shock, pale mucous membranes (Christensson, 1989), tachycardia, weak pulse, lethargy, depression, vomiting and seizures (Rockey, 1961). Usual signs are fever, anaemia, icterus, prostration, and death (Griffiths, 1978). In chronic forms of the disease, signs are vague (Ewing & Buckner, 1935). The manifestations of the disease are quite variable. Clinical findings include splenomegaly, haemoglobinuria, vomiting and diarrhoea (Gorenflot et al., 1998). Chronic babesiosis is characterised with signs such as anorexia, weight loss, lymphadenopathy, nasal discharge and bleeding tendencies. 2.8.3.1 Diagnoses, treatment and control of Babesiosis Demonstration of parasite in the erythrocyte is best accomplished by taking capillary smears from the margin of the ear (Jacobson, 2006). Giemsa stained thin and thick blood film preparations are effective in diagnosing Babesia infection (Akel & Mobarakai, 2017). Serological and molecular tests such as PCR may detect several species of Babesia (Jacobson, 2006). Acriflavine and trypaflavine have both given good results for the treatment of Babesiosis. Phenamidine is less toxic and quite effective treatment in dogs (Griffiths, 1978). Doxycycline, dexamethasone and prednisolone can be administered to lessen haemolysis but the benefit in 25 University of Ghana http://ugspace.ug.edu.gh treatment of babesiosis is uncertain. Other drugs such as imidocarb (dipropionate and dihydrochloride), parvaquone and clindamycin are used to treat babesiosis. 2.9 Gastrointestinal helminths of horses In a study by Mbafor et al. (2012) in Cameroon, enteric nematodes such as Parascaris equorum, Oxyuris equi., Strongyloides spp., Trichostrongylus axei were identified in horses. These helminths are among the major causes of colic which causes mortality in horses (Reinemeyer & Nielson, 2009b). 2.9.1 Parascaris equorum P. equorum is a large nematode found in horses and other equids (Griffiths, 1978; Anderson, 2000; Epe et al., 2004; Lyons & Tolliver, 2004). Parascaris equorum is a cosmopolitan species (Von Brand, 1952; Anderson, 2000). Adult Parascaris worms live in the small intestine of the horse (Griffiths, 1978; Reinemeyer, 2009a). The worm has three large lips, with each lip having a labial sinus on the lateral margins (Griffiths, 1978). Eggs are sub-globular and has thick, pitted, albuminous shell with brownish colour and contains a one-celled zygote between 90-100 microns in size (Griffiths, 1978). In the life cycle of P. equorum, the female worm lays eggs which develop to the first and second larval stage. The first stage larva migrates from the small intestines into the blood vessels and from there travel to the liver, where they moult into second larval stage (Clayton & Duncan, 1977). The second larval stage migrates to the lungs and invade the alveoli. At this point they are coughed up and swallowed and the larva migrates and develops into adult roundworms in the 26 University of Ghana http://ugspace.ug.edu.gh small intestines. (Griffiths, 1978). Eggs are shed in faeces and are picked up by horses in pastures. Resulting pathogenesis is especially important in young animals and may include pneumonia, bronchial haemorrhage, colic, and intestinal disturbances resulting in morbidity (Southwood et al., 1998; Schmidt et al., 2009). Intestinal perforation or obstruction is common (Cribb et al., 2006; Tatz et al., 2012). Colic results from intestinal obstructions caused by a large number of parasites (Tatz et al., 2012). Parascariasis is a common disease among horses less than one year (Austin et al., 1990). Older horses are usually immune to infection (Roberts & Janovy, 2000). Attempts to demonstrate transmammary or prenatal transmission have not been successful (Anderson, 2000). 2.9.1.1 Diagnosis, treatment and control Parascariasis Diagnosis is accomplished by finding typical brown sub-globular eggs in faecal flotation. Effective treatments include drugs such as ivermectin or moxidectin which kills the early larval stages, febendazole, pyrantel pamoate, piperazine, dichlorvos and trichlorfon (Poynther & Hughes, 1958; Lyons et al., 1976; Lyons et al., 2008a; Cobb & Boeckh, 2009). However, impaired efficacy of ivermectin and pyrantel against P. equorum and other strongyles have been found in foals (Brazik et al., 2006; Lyons et al., 2008b; Nareaho et al., 2011). Occassional resistance to piperazine has also been reported (Drudge et al., 1983). 27 University of Ghana http://ugspace.ug.edu.gh 2.9.2 Strongylus spp. Strongylus species of horses include four main species; Strongylus edentatus, Strongylus equinus, Strongylus vulgaris and Strongylus asini. These species are worldwide except Strongylus asini (Melhorn, 2015). Strongylus vulgaris is the most pathogenic Strongylus species in equids (Griffiths, 1978). The adult worm inhabits the large intestine and caecum. Strongylus vulgaris is brownish or reddish if blood has been ingested recently. The egg of S. vulgaris is 74-95 × 36-58 µm, S. edentatus egg measures 64-99 × 40-57 µm, S. equinus measures 70-90 × 40-55 µm and S. asini measures 75 × 45 µm (Melhorn, 2015). In Canada, S. vulgaris have been found in more than 85% of horses (Slocombe & McGraw, 1973). In a study by Poynther (1969), the occurrence of infection in yearlings was greater than the incidence recorded in foals The life cycle varies for the different species of Strongylus. The life cycle of S. vulgaris will be emphasized since it is the most pathogenic species. Ingested infective third stage larvae penetrate the intestinal wall and mature into fourth stage larvae in about 10 days to two weeks (Ogbourne, 1973). These larvae migrate and penetrate the intima of the submucosal arterioles, and migrate in these vessels toward the anterior mesenteric artery. About 45 days after initial infection, the fourth stage larvae return via the arterial system to the submucosa of the caecum and colon where they become fifth stage larvae about 3 months after initial infection. The larva enters the lumen, mature and produce eggs about 180-200 days after initial infection. Some larvae remain as fourth stage or fifth stage larva in aneurysms in the cranial mesenteric artery. Transmission is through the ingestion of third stage larvae with food and water. Most infective larvae die after a few months’ exposure on pasture, but some may survive as long as a year (Griffiths, 1978). There is no evidence of prenatal infection (Drudge et al., 1966) 28 University of Ghana http://ugspace.ug.edu.gh Diarrhoea is a more frequent sign of infection with strongyles (Dunn, 1969). Anaemia, emaciation, poor coat and poor performance are frequently attributed to large strongyles in the intestine. Following experimental infection in nine months old pony foals, an acute reaction characterised by a marked increase in temperature, loss of appetite, rapid loss of body weight, depression and recumbency, abdominal distress, constipation and occasionally, intermittent diarrhoea were observed (Drudge et al., 1966). 2.9.2.1 Diagnosis, treatment and control of Strongylosis Diagnosis is by demonstrating eggs from faeces, but larvae are required for specific identification. Rectal palpation for aneurysms involving the cranial mesenteric artery may be helpful in diagnosis (Griffiths, 1978). Thiabendazole is widely used and several other anthelminthic drugs have been developed for use in adult horses, including benzimidazole compounds (Bello et al., 1973; Bradley & Radharkrishnan, 1973), tetrahydropyrimidines (Cornwell et al., 1973) and organic phosphorus compounds (Cook , 1973) 2.10. Equine haemoparasites The most economically important haemoparasites of horses are Trypanosoma vivax, T. brucei Babesia bigemina, B. bovis, B. equi, B. caballi, Anaplasma and Ehrlichia (Leeflang & Ilemobade, 1977). Haemoparasites such as Babesia spp. and microfilariae have also be reported in horses in Nigeria (Pam et al., 2013; Oladipo et al., 2015). African animal Trypanosomiasis, Babesiosis and Cowdriosis caused by Amblyomma spp. are considered the most important constraints to horse rearing in sub Saharan Africa (Ajeyi & Fabiyi, 1983; Food and Agriculture Organization, 1992). 29 University of Ghana http://ugspace.ug.edu.gh Blood-borne parasites have severe and debilitating effects on the health and perhaps the reproductive success of horses. Equine haemoparasites such as Babesia are intraerythrocytic or extraerythrocytic protozoan parasites that cause equine piroplasmosis. Piroplasmosis was first reported in South Africa and was originally called anthrax fever (Henning, 1956; Theiler, 1901). The parasite that causes the disease was later renamed as Piroplasma equi (Laveran, 1901). Two different species that infect horses have been identified (Koch, 1904). The species were named Babesia caballi and Babesia equi (Nuttal & Strickland, 1910; Laveran, 1901). Babesia equi has been renamed as Theileria equi due to similarities with each other (Mehlhorn & Schein, 1998). Equine piroplasmosis was originally endemic to the Asian continent (Friedhoff et al., 1990), however international transport of horses has made equine Piroplasma a cosmopolitan parasite (Bruning et al., 1997). The parasite is now endemic in Europe, Africa, and America (Friedhoff et al., 1990; de Waal 1992; Yin et al., 1997; Camacho et al., 2005). Transmission of the parasite is facilitated by tick vectors such as Dermacentor, Rhipicephalus and Hyalomma (de Waal, 1992). Transplacental infection of foals in utero is a common cause of equine abortion (de Waal and van Heerden 2004; Phipps & Otter, 2004) Globally, piroplasmosis of horses serves as a significant threat in international horse trade and sport. (Sluyter, 2001; Irby, 2002). 2.10.1 Diagnoses, treatment and control of Piroplasmosis Diagnosis is accomplished by several techniques such as identification of parasites in giemsa- stained thin and thick blood films, culture techniques, PCR and serological assays. Examination of blood film is the gold standard for detecting and identifying haemoparasites of horses (de Waal and van Heerden, 2004). When parasitaemia is low, thick blood film smears are suitable for 30 University of Ghana http://ugspace.ug.edu.gh examining blood (de Waal & van Heerden, 2004). In vitro culture procedures provide specific and sensitive alternate approaches for examining blood samples when microscopy and serology yield unconvincing negative results for B. equi and B. caballi (Holman et al., 1993; de Waal & van Heerden, 2004; Holman et al., 1994; Zweygarth et al., 1997. In vitro culture techniques are expensive and require expertise. Diminazene is administered via intramuscular inoculation against B. caballi but is ineffective against T. equi infections (de Waal, 1992). Imidocarb is also a drug of choice and it is very effective against B. caballi and T. equi (Belloli et al., 2002). Euflavine and parvaquone are effective against both B. caballi and T. equi infections (Kuttler et al., 1987). Regular administration of acaricides can lead to a reduction in the incidence of piroplasmosis by eliminating tick vectors (de Waal, 1992). 31 University of Ghana http://ugspace.ug.edu.gh CHAPTER THREE 3.0 MATERIALS AND METHODS 3.1 Study area and population The study on dogs was conducted in one suburb (Madina) of the La-Nkwantanang Municipal Assembly of the Greater Accra Region and Nsawam of the Akwapim South Municipal District of the Eastern Region of Southern Ghana. The Greater Accra Region is second most populated region in Ghana (Ghana Statistical Service, 2012) and is reported to have the highest density of owned dogs (Veterinary Service Directorate, 2009). The study was conducted in four communities: Zongo, Washington, Redco and Libya quarters in Madina. Madina has a population of about 111,926 people, representing 2.8% of the region's total population (Ghana Statistical Service, 2014) and has coordinates 5o41’0” N, 0o10’0” W. The La Nkwantanang municipality is situated within the dry climatic zone of Ghana. The municipality experiences rainfall of 700 mm in the first rainy season and 770 mm in the second rainy season (Dickson et al., 1988). The land area of the Municipality consists of plains scattered with rippling landscape. The vegetation is dominated by shrub lands and grassland. (Ghana Statistical Service, 2014). The main economic activities in the La Nkwantanang Madina Municipality are commerce and agriculture (Ghana Statistical Service, 2014). Nsawam is the capital of the Akuapim South Municipal District in the Eastern Region of Ghana. It is located on the Accra-Kumasi highway and it is surrounded at the west by the Densu River. The study was conducted in five different communities namely: Adoagyiri, Achikorpe, Oparekrom, Akwamu and Ntoaso. The population of Nsawam Adoagyiri Municipal, according to the 2010 Population and Housing Census, is 86,000 representing 3.3% of the Eastern region’s 32 University of Ghana http://ugspace.ug.edu.gh total population (Ghana Statistical Service, 2014). The Municipality is situated between latitude 5’.45 N and 5’.58 N and longitude 0.07’W and 0.27’ W. The Nsawam-Adoagyiri Municipal covers an area of about 175 km2, out of the total area of the Eastern Region (Ghana Statistical Service, 2014). The majority (28.7%) of the employed population are engaged as service and sales workers followed by those who work as agricultural, forestry and fishery workers (22.4%). Average annual rainfall is between 125 cm and 200 cm (Ghana Statistical Service, 2014). The rainy season is usually between May to June (Ghana Statistical Service, 2014). Horses were sampled from two stables in the Greater Accra Region of Ghana. One stable was located on the Abossey Okai - Korle bu mortuary road in a locality called Ayigbe town with coordinate N 5033'8.4132" W 00 13'49.44" The total number of horses at the stable during the time of study was twenty-two. The stable has several breeds of horses including horses from Chad, Nigeria, Argentina, and Nigeria. The other stable was the Accra Polo grounds (N 5037' 3.3528" W 0010' 22.2456") located in the vicinity of Shangri-la, airport area on the Spintex road, Accra. The geographical coordinates are. The Polo ground has stables for about 108 horses and has several exotic horse breeds. The Polo grounds has membership of varied nationalities. The club offers squash, a recreational facility, with fully equipped squash court and trainer. The polo ground has riding school for people who want to learn horse riding. 3.2 Study design The study began with a reconnaissance survey of the study areas. In selecting the study area, two conveniently selected regions in Southern Ghana (Eastern and Greater Accra Regions) were clustered into municipalities. Two municipalities (La- Nkwantanag Municipal Assembly in the Greater Accra in Region and Akuapim South Municipal District in the Eastern Region) were 33 University of Ghana http://ugspace.ug.edu.gh randomly selected. Two suburbs, Madina and Nsawam in the Greater Accra Region and Eastern region respectively were then conveniently selected from the municipalities because they were thought to reflect the assumed differences in socio-cultural and socio-economic aspects and pattern of dog ownership and also because of their accessibility and proximity to the laboratory. The selection of study area for horses was strictly based on convenience since horses were known to be available in two areas in the Greater Accra Region. The research involved field sample collection and laboratory sample processing and identification of parasites. The design was a cross sectional study which was conducted from December, 2018 to May 2019. Blood samples were collected mainly from veterinary clinics and faecal samples were collected during house-to- house visits. Dogs brought to selected veterinary clinics within the period of study in selected communities were sampled for haemoparasites. A comprehensive list of house addresses of dogs in the community was obtained from records at the district veterinary clinic and veterinary field workers in Madina and Nsawam to obtain a sampling frame. Dog owning households were purposively selected to target houses where dogs were present. Dogs and dog owners were randomly sampled from the sampling frame. In houses where there was more than one dog, only one dog selected by the owner was included in the study. Horses were conveniently sampled because a sampling frame was not obtained during the time of study. The inclusion of participants in the study was strictly voluntary and based on consent of their owners, after being informed of the objectives and goals of the study. A pre-structured questionnaire was administered to dog owners to ascertain individual factors such as level of education, occupation, age, sex and pet management/husbandry practices as well as owners' awareness of zoonoses. Samples collected were sent to the parasitology laboratory at the Department of Animal Biology and Conservation Science, University of Ghana for processing. Ethical approval for the study was obtained from 34 University of Ghana http://ugspace.ug.edu.gh the Research and Ethical Review Committee of the Institutional Animal Care and Use, University of Ghana. 3.3 Estimation of sample size The sample size of dogs and horses was estimated using a formula described by (Pourhoseingholi et al., 2013; Dohoo et al., 2003). The formula used is: 𝑍2 × 𝑃 × 𝑄 𝑛 = 𝐿2 where n = required sample size, Z = Z value corresponding to the level of confidence (1.96), P = known or estimated prevalence, Q = (1-P), L = allowable error (0.05). Based on gastrointestinal helminths prevalence of 62.6% in dogs (Johnson et al., 2015) in the Greater Accra Region, P was estimated at 62.6% for dogs in this study to give maximum sample size. Hence n = 1.962×0.626×0.374/0.052 = 360. Therefore, at least 360 dogs were to be sampled for parasites. Prevalence of parasites was estimated at 50% for horses since no data on prevalence from previous work in the study area could be obtained. During the period of this study, no data had been published on haemoparasites and gastrointestinal helminths of horses in the study area. From the formula, 384 horses were to be sampled to obtain maximum sample size. 3.4 Sampling of dogs and horses Based on the sampling frame that was acquired from records at the Veterinary clinics in the study areas, a simple probability sampling technique was designed to sample dogs in the communities. Dogs were assigned numbers on pieces of paper. The numbers corresponded to the address of dogs in the communities. The pieces of paper were uniformly folded, placed in a container and mixed thoroughly. One piece of paper was drawn at a time without replacement and the container 35 University of Ghana http://ugspace.ug.edu.gh was shaken after each draw to mix the contents. Dogs that were selected by drawing lots were involved in the study. Sample size obtained from sampling frame was insufficient so convenience sampling in house-to-house visits was employed to add up to the sample size. Horses were conveniently sampled because a sampling frame was not available at the time of the study. 3.5 Classification of dogs and horses Dogs were classified as exotic breed if they were foreign and as mongrels if they were of unidentifiable breed. Dogs were classified as puppies if they were between the ages of 0-6 months, juveniles if they were between the ages of 7-12 months and as adults if they were above the age of 12 months as described by Johnson et al. (2015). Horses were classified as exotic breed if they foreign and as local breed if they originated from Ghana or West Africa. Horses were classified into less than 4 years, 4-10 years and above 10 years based on sexual maturity as described by Tesfu et al (2014) 3.6 Faecal collection, examination, processing and copromicroscopy 3.6.1 Rectal faecal extraction A gloved finger was lubricated with water-based lubricant. With the tail raised, the index finger was gently inserted into the rectum of dogs and horses. The finger was used to gently massage the rectum, curled upwards and gently withdrawn. About 5-10 g of faecal matter was placed into appropriately labelled faecal collection containers. Faecal containers were stored on ice in an ice chest. Rectal faecal extraction in puppies was done with a small size lubricated spatula. The lubricated spatula was gently inserted into the anus of well restrained puppies. The spatula was used to gently scoop out about 5-10 g of faeces into appropriately labelled faecal collection 36 University of Ghana http://ugspace.ug.edu.gh containers and stored on ice in an ice chest. In cases where rectal faecal extraction was impossible, stool samples were immediately collected from the top of fresh stool with gloved hands into appropriately labelled containers after defecation. 3.6.2 Examination of faecal sample Faeces was physically examined for colour, blood, consistency and presence of tapeworm proglottids. 3.6.3 Modified McMaster technique The Modified McMaster faecal egg count method described by (Urquhart et al., 1996; MAFF, 1986) was used to process faecal samples and quantify eggs in faeces. Faecal sample (2 g) was measured in a container on a scale. A 28 ml of NaCl flotation solution (360 g of NaCl in 1000 ml of water) was dispensed into the container, mixed and allowed to soak for 5 minutes. The mixture of faeces and flotation medium was stirred and strained into another container. Both chambers of McMaster slide were then filled with the mixture of faeces and flotation solution using a pipette. The slide was set aside for 5 minutes to allow parasite eggs to float to the surface. The slide was then observed under the microscope using 4× and 10× objective lens. All eggs inside the grid of the slide were counted using the 10× objective lens including those on the grid lines if they were greater than half of the number of eggs inside the grid in both chambers. The number of eggs in both chambers was multiplied by 50 to obtain the number of eggs per gram (epg). 3.6.4 Formol ether sedimentation technique The formol ether faecal sedimentation procedure outlined by (Cheesbrough, 2009) was used to concentrate the eggs of helminths. About 1 g (pea-sized) faeces was emulsified in 4 ml of 10% 37 University of Ghana http://ugspace.ug.edu.gh v/v formalin in a test tube using a stick. A 4 ml of 10% formalin was added to the tube, capped and mixed well by shaking. The emulsified faeces was sieved into a beaker. The suspension was transferred into a centrifuge tube and 3-4ml of diethyl ether was added. The tube was unstoppered and mixed for 15s using a vortex mixer. The tube was centrifuged at low 750-1000 g (about 3000 rpm) for 1 minute. Using a plastic bulb pipette, the plug of faecal debris was loosened from the side of the tube. The tube was upturned to remove the ether, faecal debris and formalin. The tube was returned to its upright position and the fluid from the side of the tube was allowed to drain to the bottom. The bottom of the tube was gently tapped to resuspend and mix the sediment. The sediment was transferred onto a slide, a drop of iodine was added and then covered with a cover slip. The preparation was examined at 10× and 40× objective lens. 3.7 Collection of blood and preparation of blood film A cephalic or saphenous venipuncture or ear prick blood was collected from dogs after consent had been sought from owners. The method of blood collection was decided after examining pets for veins that were accessible and convenient. Dogs were restrained with a tether and muzzle. In most cases, ear prick blood was the best choice. The puncture area was gently massaged and rubbed with 70% alcohol to force blood into the area and disinfect the puncture area. A lancet needle was used to gently pierce the ear margin and a drop of blood was placed on two frosted end glass slides, one for thin film and the other for thick film preparation. In cases where intravenous blood collection was possible, about 2 ml of intravenous blood was collected with a sterile 21-gauge hypodermic needle attached to a 5 ml syringe into appropriately labelled EDTA tubes for further analysis, and a drop on appropriately labelled glass slides. With a piece of cotton wool, the puncture area was firmly pressed to stop bleeding. Thin and thick blood film as 38 University of Ghana http://ugspace.ug.edu.gh described by (Cheesbrough, 2009) was quickly made on appropriately labelled glass slide, air dried and stored in a slide box. 3.7.1 Preparation of thin and thick film A drop of blood (about 5 µl) was placed on the end of a clean dry frosted end slide labelled with appropriate information. Another drop of about 5 µl blood was placed on the middle of the slide. A spreader slide was drawn back to touch the drop of blood at the middle of the slide and the blood was allowed to spread along the edge of the spreader. The drop of blood was spread to make a thin film. The other drop of blood was spread with an applicator stick to make a thick smear. The film was left to dry and then stored in a slide box. 3.7.2 Fixing and staining of slides Thin films prepared on the field upon return to the laboratory were fixed by dipping the thin films in methanol for 2 minutes and allowed to dry on a rack. The slides were placed on a staining rack and the methanol-fixed thin films and unfixed thick films were stained with approximately 0.5 ml 10% Giemsa stain for 10 minutes. The stain was washed off with water and left to dry. 3.7.3 Blood microscopy Thin and thick blood films were observed under the compound light microscope. The blood films were examined first with 10× and 40× objective to check the staining, morphology of the cells and parasitized cells. A drop of immersion oil was applied to film and spread to cover most of this part of the film and examined under 100× objective lens to identify parasites. 39 University of Ghana http://ugspace.ug.edu.gh 3.8 Assessment of pet management practises and owners’ knowledge on zoonosis Pre-structured questionnaires were administered to dog owners and horse stable keepers who consented to be interviewed. The questionnaire utilized primary multiple-choice questions (Appendix 1) and was designed to ascertain socio demographic information of owners, purpose of keeping dogs, respondent’s knowledge on canine or equine zoonoses and source of information, pet management practices including housing status, deworming status, nutrition, veterinary and health care, and presence of other domestic animals. 3.9 Parasite identification Identification of helminth eggs was done using morphological characteristics described by (Griffiths, 1978; Soulsby, 1982; MAFF manual, 1986; Anderson, 2000; Lichtenfels et al., 2008). Ancylostoma caninum was identified using morphometrics and number of blastomeres described by Lucio-Forster et al. (2012) and Ehrenford (1953). Trichostrongylus was identified using information from Issarapong et al. (2013). Spirocerca lupi was identified using information from Jan (2013). Identification of Babesia was done using information provided by Akel & Mobarakai (2017) and Griffiths (1978). 3.10 Statistical analyses Data was analysed using Statistical Package for Social Scientists (SPSS) Software Package, version 23. The results for continuous variables such as eggs per gram were expressed as means and as numbers and percentages of participants for categorical data such as infection status, age, and sex. Pearson's chi-square test of association and Fisher's exact test, where appropriate, was used to assess the association between risk factors and helminth infection status in dogs and horses. Kolmogorov Smirnov test of normality was used to test for normality of eggs per gram 40 University of Ghana http://ugspace.ug.edu.gh (epg). The intensity of eggs was compared among risk factors using Kruskal Wallis test. A p value less than 0.05 was considered statistically significant. 41 University of Ghana http://ugspace.ug.edu.gh CHAPTER FOUR 4.0 RESULTS 4.1 Prevalence of helminth infection in dogs A total of 285 out of 428 dogs were infected with at least one species of gastrointestinal helminth. The overall prevalence of helminth infection was 66.6%. Ancylostoma caninum eggs (Image 1) were found in 169 dogs with a species-specific prevalence of 39.5%. Dipylidium caninum proglottids and some eggs (Image 2) were found on stool samples of 112 dogs, representing a species-specific prevalence of 26.2%. Spirocerca lupi eggs (Image 3) were found in 13 dogs from Madina, representing a species-specific prevalence of 3.0% as indicated in Figure 1. Toxocara sp. ova (Image 4) were found in 171 dogs, which represented a species-specific prevalence of 40%. The overall species-specific prevalence is shown in Figure 1. The species-specific prevalence among sex, age and breed of dogs is indicated in Figure 2. 45 40 35 30 25 20 15 10 5 0 Toxocara sp Ancylostoma Dipylidium caninum Spirocerca lupi caninum Helminth parasites Figure 1: Prevalence of gastrointestinal helminth infections in dogs from Nsawam and Madina. 42 Percentage of infected dogs University of Ghana http://ugspace.ug.edu.gh 50 45 40 35 30 25 20 15 10 5 0 Male Female Exotic Mongrel 0-6 months 7-12 Above 12 months months Toxocara sp. Ancylostoma caninum Dipylidium caninum Spirocerca lupi Figure 2: Species-specific parasite prevalence among sex, breed and age of dogs. 4.2 Effect of age, sex and breed of dog on helminth species prevalence Toxocara, Ancylostoma and Dipylidium infections were not statistically associated with age, sex and breed of dog (p ˃ 0.05). S. lupi infection was significantly associated with breed of dog (p ˂ 0.05). The results are summarised in Table 1. 43 Percentage of infected horses University of Ghana http://ugspace.ug.edu.gh Table 1: Chi-square and Fisher’s exact test of association of helminth infection with age, sex and breed of dog Variable Toxocara A. caninum D. caninum S. lupi P 𝜒2 p P 𝜒2 p P 𝜒2 p P Fisher’s value value value p value Age (months) 0-6 33.7 38 20.7 3.3 1.000 7-12 43 0.498 37.3 0.416 26.1 0.336 2.8 ˃ 12 40.7 41.8 29 3.1 Sex Male 39.9 0.880 39 0.809 28 0.516 4.1 0.261 Female 40 40 25 1.9 Breed Exotic 39.3 0.762 41 0.343 24.4 0.582 7.4 0.001 Mongrel 40.3 39 27 1.0 4.3 Intensity of helminth infection in dogs The overall mean intensity of Toxocara was 2106 ± 159 and the mean intensity of Ancylostoma caninum was 4653 ± 214. The mean intensities of helminth infection among age, sex and breed of dogs are summarized below in Table 2. The results indicate the difference in mean intensities was not statistically significant (p ˃ 0.05) as indicated in Table 2. 44 University of Ghana http://ugspace.ug.edu.gh Table 2: Intensity of helminth infection among age, sex and of dogs Variable Toxocara Ancylostoma caninum Mean intensity Kruskal- Mean Kruskal- Wallis p intensity Wallis p value value Age (months) 0-6 2051 ± 342 5014 ± 522 7-12 1726 ± 220 0.498 3989 ± 351 0.416 ˃ 12 2416 ± 265 4933 ± 306 Sex Male 2043 ± 224 0.880 4695 ± 323 0.809 Female 2173 ± 228 4610 ± 283 Breed Exotic 2635 ± 338 0.762 5284 ± 377 0.343 Mongrel 1871 ± 171 4348 ± 257 4.4 Risk factors affecting prevalence of helminth infection in dogs A chi-square analysis of the risk factors associated with helminth infection in dogs revealed that only deworming status as a management practice was significantly associated with helminth infection (p ˂ 0.05). All other variables (risk factors) were not statistically associated with helminth infection. Among the age groups, prevalence was higher in dogs above 12 months but the result was statistically insignificant. Among sex, female dogs recorded higher helminth prevalence than male dogs. The results however, was statistically insignificant (p ˂ 0.05). Helminth prevalence was higher in exotic dog breeds, semi-restricted dogs and dogs that were fed both homemade and commercial food. In all cases, the results were statistically insignificant. The results of the analysis are summarised in Table 3. 45 University of Ghana http://ugspace.ug.edu.gh Table 3: A chi-square analysis of some factors associated with overall helminth prevalence in dogs Variables Total Number of dogs 𝜒2 p value (428) infected (%) Age 0-6 months 92 56 (60.9) 1.897 0.387 7-12 months 142 95 (66.9) ˃ 12 months 194 134 (69.1) Sex Male 218 140 (64.2) 1.120 0.290 Female 210 145 (69.0) Breed Exotic 135 94 (69.6) 0.820 0.365 Mongrel 293 191 (65.2) Deworming status Yes 122 72 (59.0) 4.398 0.036 No 306 213 (69.6) Mobility of dog Restricted 247 158 (64) 1.804 0.179 Semi-restricted 181 127 (70.2) Food type Kibble 19 11 (57.9) 2.173 0.337 Homemade 375 248 (66.1) Both 34 26 (76.5) a. Restricted dog were dogs that were confined to a house and were not allowed to freely roam outside without supervision b. Semi-restricted dogs (Neighbourhood dogs) were dogs that roamed freely outside usually without supervision and were dependent on owners for food and shelter. 46 University of Ghana http://ugspace.ug.edu.gh Image 1. A micrograph of Ancylostoma caninum eggs viewed under ×400 magnification. Source: Baah (2019) Image 2. A micrograph of Dipylidium caninum egg packet viewed under ×400 magnification. Source: Baah (2019) 47 University of Ghana http://ugspace.ug.edu.gh Image 3. A micrograph of Spirocerca lupi ova under ×400 magnification. Source: Baah (2019) Image 4. A micrograph of a one cell stage of Toxocara sp. viewed under ×100 magnification on a McMaster slide. Source: Baah (2019) 48 University of Ghana http://ugspace.ug.edu.gh 4.5 Prevalence of helminth infections in horses A total of 29 out of 73 horses were infected with at least one species of gastrointestinal helminth. The overall prevalence of helminth infection was 39.7%. Strongyle-type eggs (Image 5) were found in the stool of 19 horses representing a prevalence of 26.0%. Trichostrongylus sp. eggs (Image 6) were identified in 6 horses, representing a prevalence of 8.2%. Parascaris sp. eggs (Image 7) were found in the stool of 24 horses, which represented a prevalence of 32.9%. The prevalence of helminths in horses is summarized in Figure 3. The species-specific prevalence among age, sex and breed of horses is shown in Figure 4. 35 30 25 20 15 10 5 0 Parascaris sp. Strongyle-type egg Trichostrongylus Helminth parasites Figure 3. Prevalence of gastrointestinal helminths in horses. 49 Percentage of horses infected University of Ghana http://ugspace.ug.edu.gh 50 45 40 35 30 Parascaris 25 Strongyle-type egg 20 Trichostrongylus 15 10 5 0 Male Female Exotic Local ˂ 4 years 4-10 years˃ 10 years Figure 4: Species-specific prevalence of gastrointestinal helminths among age, sex and breed horses. 4.6 Factors affecting the prevalence of helminth infection in horses Management practices like deworming status, frequency of deworming and even food type that affect infection were same in all the two horse stables. Hence the only variables that could be used to explain helminth infection status were inherent variables like age, sex, breed, and stables. The results are summarised in Table 4. 50 Percentage of horses infected University of Ghana http://ugspace.ug.edu.gh Table 4: A chi-square test of association of some factors that affect prevalence of helminths in horses Variables Total Number of dogs 𝜒2 p value (73) infected (%) Age (years) ˂ 4 36 18 (50) 3.468 0.177 4-10 23 6 (26.1) ˃ 10 14 5 (35.7) Sex Male 49 18 (36.7) 0.557 0.455 Female 24 11 (45.8) Breed Exotic 47 17 (36.2) 0.697 0.404 Local 26 12 (46.2) Stables Polo grounds 51 16 (31.4) 4.932 0.026 Ayigbe town 22 13 (59.1) Image 5. A micrograph of Strongyle-type egg under ×400 magnification. Source: Baah (2019) 51 University of Ghana http://ugspace.ug.edu.gh Image 6. A micrograph of Trichostrongylus egg under ×400 magnification. Source: Baah (2019) Image 7. A micrograph of egg of Parascaris sp. under ×400 magnification. Source: Baah (2019) 52 University of Ghana http://ugspace.ug.edu.gh 4.7 Effect of age, sex and breed on species-specific helminth prevalence in horses Parascaris infection were not statistically associated with age, sex and breed of dog (p ˃ 0.05). Strongyle-type egg infection and Trichostrongylus egg infection were significantly associated with breed of dog and breed of dog (p ˂ 0.05). The results are summarised in Table 5. Table 5: Fisher’s exact test of association of age, sex and breed with species-specific helminth infection in horses. Variable Parascaris Strongyle-type egg Trichostrongylus P 𝜒2 p value P 𝜒2 p P 𝜒2 p value value Age (months) ˂ 4 36.1 39 16.7 4-10 26.1 0.757 17.4 0.041 0 0.041 ˃ 10 35.7 7.1 0 Sex Male 28.6 0.297 28.6 0.577 10.2 0.656 Female 41.7 20.8 4.2 Breed Exotic 29.8 0.603 14.9 0.005 0 0.001 Local 38.5 46.2 23.1 4.8 Intensity of equine helminth infection The mean intensity of Parascaris egg infection was 802 (range 100-2000), while the mean intensity of Strongyle-type egg was 390 (range 150-2050) and that of Trichostrongylus egg was 791 (range 350-1250) among all horses Age-, breed- and sex- specific mean intensity of helminth species in horses is shown in Table 6. 53 University of Ghana http://ugspace.ug.edu.gh Table 6: Intensity of helminth infection among age, sex and breed of horses. Variable Parascaris Strongyle-type egg Trichostrongylus Mean Kruskal- Mean Kruskal- Mean Kruskal- intensity Wallis p intensity Wallis p intensity Wallis p value value value Age (months) ˂ 4 615 ± 145 790 ± 114 791 ± 149 4-10 1025 ± 106 0.863 733 ± 109 0.009 0 0.037 ˃ 10 1020 ± 258 2050 0 Sex Male 768 ± 140 0.221 867 ± 155 0.776 820 ± 179 0.375 Female 850 ± 162 792 ± 108 650 Breed Exotic 2635 ± 338 0.659 1114 ± 207 0.005 0 0.001 Local 1871 ± 171 700 ± 118 791 ± 149 4.9 Prevalence of canine haemoparasites Out of 156 dogs that were examined for haemoparasites, 54 were from Madina and 102 were from Nsawam. Only 12 dogs were positive for haemoprotozoan parasites (7.7%). All infected dogs were adult mongrels from Nsawam. The haemoprotozoan parasites were identified as Babesia spp. (Image 8). 54 University of Ghana http://ugspace.ug.edu.gh Image 8. An image of giemsa stained blood smear showing intraerythrocytic haemoprotozoan Babesia-like parasites in red blood cells of dog. Source: Baah (2019) 4.10 Prevalence of equine haemoparasites Out of 73 horses that were examined for haemoparasites, 22 were from Ayigbe town stable and 51 were from Polo grounds stable. Eight (11.0%) horses were positive for haemoprotozoan parasites. This prevalence was recorded in adult horses from Ayigbe town stable. There were no positive cases of haemoparasites in horses from Polo grounds stable. Haemoprotozoan parasites were identified as Babesia spp. (Image 9). 55 University of Ghana http://ugspace.ug.edu.gh Image 9. A micrograph of Babesia-like haemoprotozoan parasite under oil immersion lens. A: Red blood cell with thin, delicate pyriform ring consisting of cytoplasmic rim with chromatin dot. B: Parasite appears as chromatin dot in red blood cell. 4.11 Socio-demographic characteristics of respondents A total of 428 dog owners were recruited from both study communities and included in the study (Table 7). The mean age of dog owners (standard error of mean) was 37.9 (0.5), ranging from 21- 68 years. All respondents had formal education. Trading was the commonest occupation. 56 University of Ghana http://ugspace.ug.edu.gh Table 7: Summary of socio-demographic characteristics of dog owners from the study sites. Demographic Number (%) of respondents in the study area information Nsawam Madina Total (n=245) (n=183) (n=428) Gender Male 196 (80) 145 (79.2) 341 (79.7) Female 49 (20) 38 (20.8) 87 (20.3) Age (years) 18-35 124 (50.6) 101 (55.2) 225 (52.6) ˃ 35 121 (49.4) 82 (44.8) 203 (47.4) Level of education Primary 53 (21.6) 33 (18.0) 86 (20.1) Junior High 44 (18.0) 27 (14.8) 71 (16.6) Senior High 60 (24.4) 67 (36.6) 127 (29.7) Tertiary 88 (36) 56 (30.6) 144 (33.6) Religious status Christian 233 (95.1) 170 (92.9) 403 (94.2) Muslim 12 (4.9) 13 (7.10) 25 (5.8) Occupation Farming 40 (16.3) 13 (7.1) 53 (12.4) Trading 105 (42.9) 93 (50.8) 198 (46.3) Office work 40 (16.3) 47 (25.7) 87 (20.3) Teacher 38 (15.5) 19 (10.4) 57 (13.3) Artisan 22 (9.0) 11 (6.0) 33 (7.7) 4.12 Number of owned dogs A total of 428 participants (dog owners) were interviewed in households. The number of dog owners that owned one dog was 76 (17.8%), 98 (22.9%) owned two dogs, 80 (18.7%) owned three dogs, 68 (15.9%) owned four dogs and the remaining 106 (24.8%) owned 5-9 dogs. The average number of dogs per dog owning household was 3.23 ± 1.76. A stratification by number of dogs owned in the study sites is summarised in Table 8. 57 University of Ghana http://ugspace.ug.edu.gh Table 8: Summary of number of dogs owned by dog owners from the study sites Number of owners (%) Dog per owner Madina Nsawam 1 31 (16.9%) 45 (18.4%) 2 47 (25.7%) 51 (20.8%) 3 28 (15.3%) 52 (21.2%) 4 17 (9.3%) 51 (20.8%) 5 28 (15.3%) 46 (18.8%) 6 13 (7.1%) 0 7 8 (4.4%) 0 8 6 (3.3%) 0 9 5 (2.7%) 0 Total 183 245 4.13 Factors affecting dog ownership From the survey, there were more dog owners that were males (79.7%) than dog owners that were females (20.3%). Dog owners primarily kept dogs for security reasons (89.7%) than for companionship reasons (10.3%). No dog owner kept a dog for hunting reasons. Majority of the dog owners were Christians (94.2%), whilst only a few were Muslims (5.8%). A one sample chi- square analysis indicated that the differences in percentages of owners based on gender, religion and role of dogs were statistically significant (p ˂0.001). The results are summarised in Table 9. 58 University of Ghana http://ugspace.ug.edu.gh Table 9: A one sample chi-square analysis of differences in percentages of some variables that affect dog ownership Variable Number of owners Chi-square p value (%) 𝜒2( ) Sex of owner Male 341 (79.7) 150.738 ˂ 0.001 Female 87 (20.3) Religion of owner Christian 403 (94.2) 333.841 ˂ 0.001 Muslim 25 (5.8) Role of dog Security 384 (89.7) 270.093 ˂ 0.001 Companionship 44 (10.3) Educational background Primary 86 (20.1) Junior High 71 (16.6) 32.766 ˂ 0.001 Senior High 127 (29.7) Tertiary 144 (33.6) Owner’s occupation Farming 53 (12.4) Trading 198 (46.3) 201.907 ˂ 0.001 Office work 87 (20.3) Teacher 57 (13.3) Artisan 33 (7.7) Age group of owners 18-35 years 225 (52.9) 1.131 0.288 Above 35 years 203 (47.1) 4.14 Dog management practices From the questionnaire interview, it was observed that 371 (86.7%) of the respondents (dog owners) were aware of rabies as a canine related zoonotic disease, whilst 57 (13.3%) were not aware of rabies as a canine related zoonosis. The difference in response, in terms of knowledge on rabies as a zoonosis, was statistically significant (p ˂ 0.001). When the responses were stratified into educational background of dog owners, 64 (17.3%) of primary school level dog owners knew 59 University of Ghana http://ugspace.ug.edu.gh about canine rabies, 57 (15.4 %) of Junior High School level dog owners knew about canine rabies, 117 (31.5%) of Senior High School level dog owners knew about canine rabies and 133 (35.8%) of tertiary school level dog owners knew about canine rabies. When asked about their source of awareness of canine rabies, 28.4% of the respondents said they were educated on rabies by peers, 25.2% by media sources and 46.4% by veterinary service officers. None of the respondents knew of any canine helmintho-zoonoses. Data collated on dog management practices, owners’ knowledge and attitudes with respect to zoonoses from the questionnaire survey was as tabulated below in Table 10. Table 10: Knowledge, practices and attitudes of respondents with respect to dog ownership Management practices Number (%) of respondents in the study area Nsawam (n=245) Madina (n=183) Total (n=428) Deworming status Yes 63 (25.7) 68 (37.2) 131 (30.6) No 182 (74.3) 115 (62.8) 297 (69.4) Frequency of deworming At least once every six months 10 (15.9) 1 (1.5) 11 (8.4) At most once a year 53 (84.1) 67 (98.5) 120 (91.6) Food type Commercial dog food (Kibble) 2 (0.8) 17 (9.3) 19 (4.4) Homemade food 239 (97.6) 136 (74.3) 375 (87.6) Both 4 (1.6) 30 (16.4) 34 (8) a Homemade food Gari + palm oil + other accompaniment 98 (41) 14 (10.3) 112 (29.9) Leftover household meal 102 (42.7) 87 (64) 189 (50.4) Cooked offal + other accompaniment 23 (9.6) 7 (5.1) 30 (8) Uncooked offal 12 (5.0) 21 (15.4) 33 (8.8) Frequency of feeding dog Once a day 232 (94.7) 145 (79.2) 377 (88.1) Twice a day 13 (5.3) 38 (20.8) 51 (11.9) Dog food dish Bowl 185 (75.5) 153 (83.6) 338 (79) On the ground 60 (24.5) 30 (16.4) 90 (21) Housing of dog Housed (Kennel) 49 (20) 66 (36.1) 115 (26.9) Unhoused 196 (80) 117 (63.9) 313 (73.1) Mobility of dog 60 University of Ghana http://ugspace.ug.edu.gh Restricted 158 (64.5) 89 (48.6) 247 (57.7) Semi-restricted 87 (35.5) 94 (51.4) 181 (42.3) Frequency of bathing Once a month 3 (1.2) 22 (12.0) 25 (5.8) At least twice a year 23 (9.4) 41 (22.4) 64 (15.0) Never 219 (89.4) 120 (65.6) 339 (79.2 Do you take dog out on walk? Yes 16 (6.1) 114 (62.3) 130 (30.4) No 229 (93.9) 69 (37.7) 298 (69.6) Where does dog defecate? On house compound 219 (89.4) 133 (72.7) 352 (82.2) Outside house compound 24 (10.6) 50 (27.3) 74 (17.8) Reason for keeping dogs Companionship 26 (10.6) 18 (9.8) 44 (10.3) Security 219 (89.4) 165 (90.2) 384 (89.7) Cohabitation of dogs with other domestic animals Yes 142 (58) 68 (37.2) 210 (49.1) No 103 (42) 115 (62.8) 218 (50.9) Ectoparasite control Yes 84 (34.3) 69 (37.7) 153 (35.7) No 161 (65.7) 114 (62.3) 275 (64.3) Means of ectoparasite control Removal of ectoparasites with hand 46 (54.8) 17 (24.6) 63 (41.2) Use of acaricide 38 (45.2) 52 (75.4) 90 (58.8) Owners’ awareness of canine zoonoses Yes 206 (84.1) 165 (90.2) 371 (86.7) No 39 (15.9) 18 (9.8) 57 (13.3) Canine zoonoses Rabies 206 (84.1) 165 (90.2) 371 (86.7) Helminthozoonoses 0 0 0 Protozoonoses 0 0 0 Restricted dogs were dogs that there were strictly kept in the confines of a home and were not allowed to roam outside without supervision. Semi-restricted dogs (Neighbourhood dogs) were dogs that roamed freely outside usually without supervision and were dependent on owners for food and shelter. Unhoused dogs were dogs with no shelter that roamed freely on house compound or were neighbourhood dogs that returned home on daily basis. Homemade food included a mixture of “gari” and palm oil, boiled offal or left-over food cooked for the family. 61 University of Ghana http://ugspace.ug.edu.gh Homemade food sometimes included offal from slaughtered livestock such as domestic fowl, goat and innards of cattle from slaughter houses. Table 11: Chi-square test of association of canine management practices with breed of dog Management practices Breed of dog Local breed Exotic breed Chi-square p value (Mongrel) n = 135 2 test (𝜒 ) n = 293 Deworming status Yes 42 (14.3) 89 (66) 115.817 ˂ 0.001 No 251 (85.7) 46 (34) Food type Commercial dog food 0 (0) 19 (14) (Kibble) Homemade food 293 (100) 82 (61) 131.287 ˂ 0.001 Both 0 (0) 34 (25) Frequency of feeding Once a day 279 (95.2) 98 (72.6) 45.089 ˂ 0.001 Twice a day 14 (4.8) 37 (27.4) Dog food dish Bowl 208 (71) 130 (96.3) 35.641 ˂ 0.001 On the ground 85 (29) 5 (3.7) Housing Kennel 47 (16) 68 (50.4) 55.429 ˂ 0.001 Unhoused 246 (84) 67 (49.6) Mobility of dog Restricted 150 (51.2) 97 (71.9) 16.159 ˂ 0.001 Semi-restricted 143 (48.8) 38 (28.1) Frequency of bathing Once a month 10 15 At least twice a year 14 50 92.321 ˂ 0.001 Never 269 70 62 University of Ghana http://ugspace.ug.edu.gh 4.15 Socio-demographic characteristics of stable keepers and horses 4.15.1 Stable keepers A total of 6 stable keepers were recruited and interviewed in the study. All the stable keepers were males between the ages of 12-23 years. All the stable keepers had no secondary school level education. 4.15.2 Horses The commonest horse breeds that were encountered were West African pony (local breeds) representing 26 (35.6%) of horses sampled. Forty-seven (64.4%) were exotic breeds. Thirty-six (49.3%) of the horses were less than the age of 4 years, twenty-three (31.5%) were between the ages of 4-10 years and 14 (19.2%) were above 10 years. Male horses that were sampled were 49 (67.1%), whilst female horses were 24 (32.9%). 4.16 Horse management practices From the results, all horses received anthelminthic treatment. However, other management practices were different in the stables that were studied. The results are summarised in Table 12. Table 12: Summary of data on horse management practices and percentage of horses under management practices at Ayigbe town stable and Polo grounds stable. Management practices Percentage of horses Polo grounds stable Ayigbe town stable Deworming status Yes 100 100 No 0 0 Frequency of deworming Once every three months 100 100 Veterinary care 63 University of Ghana http://ugspace.ug.edu.gh Yes 100 0 No 0 100 Food type Wheat bran + pasture 100 100 Foraging Yes 100 0 No 0 100 Housing Stall 100 0 Unhoused 0 100 Frequency of cleaning faeces in stalls or compound Once a day 100 100 Twice a day 0 0 Unhoused horses were restrained with a leash and were not allowed to freely roam on their own. 4.17 Caretakers and stable keepers’ perception on equine zoonoses From the interview of stable keepers, it was revealed that the stable keepers were unaware of any potential zoonotic infections that could be contracted from horses. Stable keepers from Ayigbe town stable reported that they sometimes picked dung of horses with their bare hands without wearing gloves or any form of protection. When asked why they picked dung with their bare hands, one stable boy said that he picked the dung when they are dry in the sun and any “germ” in the dung would already be dead. Another stable keeper responded that the dung is simply grass and it is not as foul and disgusting as the faeces of a dog. Image 10 shows an image of a stable keeper picking horse dung with his bare hands. 64 University of Ghana http://ugspace.ug.edu.gh Image 10. A stable keeper picking dung with bare hands. Source: Field survey, Baah (2019) 65 University of Ghana http://ugspace.ug.edu.gh CHAPTER FIVE 5.0 DISCUSSION 5.1 Factors and trend of dog ownership To the best of my knowledge, this study is the first of its kind to investigate the factors that influence dog ownership in Ghana and its potential implications for public health. Investigations were centred on a small study area; hence generalisations cannot be translated to the whole of Ghana. Dog owning households were randomly selected within a defined geographic area, hence minimising bias. In this study, a one sample chi-square analysis indicated that there was a highly significant difference in percentages of male and female dog owners. More males owned dogs than females. This is probably because the head of most Ghanaian households are mostly males. This is in agreement with a report by Knobel (2008) who reported more dog owners who are males than dog owners who are females. This is contrary to a study by Westgarth et al.( 2007) who reported more dog owners who are females in households and concluded that adult females were positively associated with dog ownership in UK. Similary, Wise & Kushman (1984) reported that females were more likely to own dogs than males. The difference possibly reflects the socio-cultural and socio-economic factors that affect dog ownership in these countries. Among religion, there was significantly lower number of muslim dog owners compared to Christian dog owners in this study. The difference can be attributed to the perception of dogs in the Islamic religion. In Islam, dogs are considered impure in a spiritual sense, although dog ownership is not prohibited (Foltz, 2005; Beck, 2000). Knobel et al. (2008) also found significant 66 University of Ghana http://ugspace.ug.edu.gh association between religion and dog ownership where few dog owners were muslims than christians in Tanzania. Dog ownership pattern reported in this study is consistent with Knobel, 2008 who reported a high percentage of dog owners (61.9%) that kept dogs for security reasons in Tanzania. However, this is contrary to the findings of Leslie et al. (1994) who reported that the highest scoring reason for dog ownership was companionship in a study in Canada. Other studies have reported that the reason for owning a dog in the western world is companionship (Staats et al., 2008; Bennett & Rohif, 2007). In this study, age of owner did not significantly influence dog ownership. This finding was in contrast to (Knobel, 2008) who reported that households headed by people within 16-35 years were significantly less likely to own dogs than household headed by middle-aged individuals. Owner’s level of education and occupation have been considered as a proxy for socio-economic status (Holland, 2019). A study by Murray et al. (2010) reported that the likelihood of dog ownership decreased with increasing educational level. Murray and colleagues’ findings is somewhat contrary to the findings in this study. In this study, a one sample chi-square analysis revealed that there were more tertiary education level dog owners. The reason for this observation could be of socio-economic or socio-cultural status. It is likley that tertiary education level dog owners have the financial resources to cater for a dog. The effect of owner’s occupation has been variably reported with regard to the likelihood of dog ownership. In a multivariable analysis in the UK (Westgarth et al., 2007) and Canada (Leslie et al., 1994), the effect of owner’s occupation was not significantly associated with dog ownership. Dog ownership has been reported to be associated with high income (Franti & Kraus, 1974). 67 University of Ghana http://ugspace.ug.edu.gh Differences in dog ownership pattern with respect to income level (Leslie et al., 1994; Franti & Kraus, 1974) are possibly comparable to variations in owner’s occupation (Westgarth et al., 2007). In this study, a one sample chi-square analysis revealed that the were more dog owners who were traders and most traders (market sellers, artisans, etc.) are not expected to receive high income. The differences between findings of these studies might be explained by the different measures used to access occupation and income. The variability in findings also suggest that these factors may also be inter-dependent with other influences (Holland, 2019). Generally, differences in dog ownership pattern in Ghana compared to patterns in other parts of the world may be due socio-cultural variations and owners’ demographics. 5.2 Prevalence of gastrointestinal helminths in dogs The overall prevalence of gastrointestinal helminths in dogs in this study was 66.6%, which was similar to the prevalence reported by Johnson et al. (2015) in Greater Accra Region of Ghana (62.6%), Ugbomoiko et al. (2008) in Nigeria (68.4%). However, the prevalence in this study was higher than the prevalence reported by Amissah-Reynolds et al. (2016) in Mampong, Ashanti region of Ghana (52%), Wyckliff et al. (2017) in Kenya (35.29%), Anene et al. (1995) in Nigeria (37.6%), Okoye et al. (2011) in stray dogs in Nigeria (52.6%), Sowemimo & Ayanniya, (2017) in Nigeria (41.7%). The prevalence in this study was lower than the prevalence reported by Tamerat et al. (2015) in Ethiopia (91%), Komatangi (2005) in Cameroon (88.5%), Abere et al. (2013) in Ethiopia (84.78%). The prevalence of helminth parasites varies significantly from one locality to another depending on the helminth species involved, the host species, management practices, anthelminthic treatment, diagnostic procedures use and local climatic conditions such as humidity, temperature, 68 University of Ghana http://ugspace.ug.edu.gh rainfall (Katagiri & Oliveira-Sequeira, 2008; Oliveira-Sequeira et al., 2002; Robertson et al., 2000). The poor veterinary attention given to owned dogs in this study is emphasized when the higher prevalence of 66.6% in this study is compared to the lower prevalence of 52.6% in stray dogs in Nigeria reported by Okoye et al. (2011). Also, Amissah-Reynolds et al. (2016) and Wyckliff et al. (2017) employed a single flotation technique which may have underestimated the prevalence since a combination of methods have been reported to increase the chances of recovering more parasites (Zewdu et al., 2010). The helminths indentifed in this study were Ancylostoma caninum, Toxocara sp., Dipylidium caninum and Spirocerca lupi. These helminths have been identified and reported in Ghana (Amissah-Reynolds et al., 2016; Johnson et al., 2015; Anteson & Corkish, 1975) and other parts of the world with geographical, ecological, epidemiological and seasonal variations in prevalence (Alvarado-Esquivel et al., 2015) in Mexico, Gracenea et al. (2009) and Cantó et al. (2011) in Spain, Ngui et al. (2014) in Malaysia, Kutdang et al. (2010) in Nigeria, Davoust et al. (2008) in Gabon and Minnaar & Krecek (2001). In the works done by Amissah-Reynolds et al. (2016) and Johnson et al. (2015), a single flotation technique with sodium chloride as a flotation medium was used and Spirocerca lupi was not reported. Reports by Fox et al. (1988), Evans (1983) and Traversa et al. (2008) indicated that classical flotation methods are insensitive diagnostic techniques for Spirocerca lupi. This may explain why S. lupi was not identified in their study. It is also possible that the dogs were not infected with S. lupi. In this study, a low prevalence of 3.0% was reported for S. lupi. According to Christie et al. (2011), the reported sensitivity for the detection of S. lupi eggs in faeces is low 69 University of Ghana http://ugspace.ug.edu.gh and was highest when centrifugal flotation technique with NaNO3 was used. Spirocerca lupi in this study however, were recovered using the formol-ether sedimentation technique. This is in contrast to (Markovics & Medinski, 1996) who recorded no S. lupi egg using faecal sedimentation tests. Faecal samples sometimes require repeated examinations when the first examination is negative for S. lupi since the shedding of eggs is often intermittent (Dvir et al., 2010). It is likely that Spirocera lupi was underestimated in this study considering samples were analysed once and that floatation and sedimentation techniques are ineffective for S. lupi diagnosis (Minnaar & Krecek, 2001). The findings in this study agree with Amissah-Reynolds et al. (2016) in Ghana, Ugbomoiko et al. (2008) in Nigeria and Kimura et al. (2013) in Japan who reported Toxcara as the most prevalent helminth. In this study, a prevalence of 40% was recorded for Toxocara spp. Toxocara is a cosmopolitan soil-transmitted helminth and hence habits like feeding off the floors and sleeping on bare grounds may account for this observation (Amissah-Reynolds et al., 2016). Toxocara has sticky eggs and eggs may stick to the coat of dogs when they sleep on the bare ground (Kleine et al., 2017; Rapini et al., 2012). Infection may occur when dogs groom their fur by licking them. Species identification of Toxocara by microscopy was impossible since micrographs of the eggs on McMaster slide could only be viewed under ×10 objective lens. This power of lens could not present clear details for identification to species level. It is known that dogs can harbour both dog and cat Toxocara spp. (T. canis and T. cati) (Anderson, 2000). Considering that dogs are coprophagous and may live with cats in the same household, brings up the possibility that some of the Toxocara eggs found in this study could be feline Toxocara cati. A more sensitive molecular technique like PCR (Polymerase chain reaction) can distinguish between the two Toxocara species. 70 University of Ghana http://ugspace.ug.edu.gh This study also recorded a lower prevalence of D. caninum infection than already reported prevalence by Johnson et al. (2015) in Ghana. However, prevalence of Dipylidium caninum was higher than the prevalence reported in Poland (Tylkowska et al., 2010) and Ghana (Amissah- Reynolds et al., 2016). Postmortem examination of dogs by Zewdu et al. (2010) and Anteson & Corkish, (1975) revealed significantly higher prevalence than the prevalence reported from copromicroscopy in this study. It is believed that D. caninum eggs are focally distributed and are rarely seen in stool due to the intermittent nature of proglottid shedding (Barnett et al, 2013), hence necropsy is more effective in recovering D. caninum (Minnaar & Krecek, 2001). It is therefore likely that D. caninum infection in this current study was underestimated considering the method of diagnosis that was used. In this study, there was no significant difference in the overall helminth infection among age groups of dogs. This finding is in agreement with reports from Abere et al. (2013), Cantó et al. (2011), Alvarado-Esquivel et al. (2015) and Awoke et al. (2011). However, reports from Ehimiyein et al. (2018), Idika et al. (2017), Amissah-Reynolds (2016), Tamerat et al. (2015), Johnson et al. ( 2015), Panigrahi et al. (2014), Okpara et al. (2015), and Adamu et al. (2012) stated otherwise. Authors like Johnson et al. (2015) and Tamerat et al. (2015), Swai et al. (2010) have reported significantly higher prevalence in puppies. However, some authors used different age group classifications from the one that was used in this study, hence age-wise prevalence comparisons may not be valid. This study reported no statistical significance in the difference between Ancylostoma caninum, Toxocara spp., Dipylidium caninum and Spirocerca lupi infections among age groups. Senlik et al. (2006), Zelahem & Mekonnen (2012), Overgaauw & Boersema (1998), Katagiri & Oliveira- Sequeira (2008), Swai et al. (2010) and Ferreira et al. (2016) reported otherwise. Toxocara and 71 University of Ghana http://ugspace.ug.edu.gh Ancylostoma infections in puppies have been attributed to transmammary route through milk and transplacental transmission in utero (Burke & Robertson, 1985; Gillespie, 1988; Anderson, 2000). Dipylidium infections are acquired through the ingestion of infected flea (Neira et al., 2008; Ramana et al., 2011). Dipylidium infection in puppies is therefore attributable to ingestion of infected fleas from their mothers when puppies lick their fur or when they accidentally ingest infected flea whilst breastfeeding. There was no significant difference in helminth prevalence among sexes in this study. Female dogs had a higher helminth prevalence than male dogs, but the difference was not statistically significant Other researchers have reported similar findings. Idika et al. (2017) reported higher prevalence in male dogs but did not report statistical significance. This is in agreement with reports by Amissah-Reynolds et al. (2016), Tamerat et al. (2015), Wyckliff et al. (2017), Abere et al. (2013), Jones et al. (2011), Awoke et al. (2011) and Ehimiyein et al. (2018). However this finding is in contrast to reports by Adamu et al. (2012), Okpara et al. (2015) and Alvarado- Esquivel et al. (2015) who reported significant differences in infection among sexes. There was no significant association between breed of dog and helminth prevalence in this study. Significant difference in prevalence among breeds have been reported by Idika et al. (2017) in Nigeria. A number of reports have also reported significant association among breeds with local dogs (non-exotic breeds) having higher helminth prevalence (Abere et al., 2013, Idika et al., 2017; Awoke et al., 2011). In Nigeria, Onyenwe & Ikpegbu (2004) reported a significantly higher prevalence in cross breeds over the local and exotic breeds. However, a report by Anosike et al. (2004) and Swai et al. (2010) found no statistically significant association between breed and helminth infection. The high prevalence of infection in local dogs is probably due to the different management practices given to local dogs. In this study, it was found that management practices 72 University of Ghana http://ugspace.ug.edu.gh such as deworming, food type (kibble or homemade food), dog housing (kennel or unhoused), mobility of dog (semi-domestic or strictly domestic), dog food dish (dog bowl or ‘’fed off the ground’’) and even frequency of bathing differed significantly among dog breeds. It is suspected that because exotic dog breeds are expensive, they are appreciated and given proper veterinary and husbandry care with little or no roaming range outside of their homes. On the contrary, local dogs are cheaper, common, less appreciated and are allowed to freely roam and either become communal, semi-domestic or stray dogs where they are most likely to harbour infections from other dogs or from the food they eat from the streets (Idika et al., 2017; Aiyendun & Olugasa, 2012). A greater proportion of dog owners with low socio-economic status who owned local dogs did not exercise proper veterinary and management practices. They exposed their dogs to infections by allowing unrestricted maneuverability of their dogs. Resources and education were lacking and owners may exercise veterinary care only when the dog is obviously sick, hence the high prevalence in local dogs. Local dogs that were fed ‘gari and palm oil’were emaciated and look malnourished. According to Taylor (2007), heavy helminth infection in malnourished dogs cause anaemia and protein loss. A highly significant association was found between deworming status and helminth prevalence in this study (Table 8). This is in line with reports from Alvarado-Esquivel et al. (2015) in Mexico and Satyal, et al. (2013) in Nepal. Other authors however, have found no significant association between deworming and helminth prevalence. Johnson et al. ( 2015) in Ghana, Wyckliff et al. (2017) in Kenya and Swai et al. (2010) in Tanzania did not find any significant association between deworming and helminth prevalence in dogs. In Johnson and colleagues’ work, the sample size of dewormed and undewormed dogs was low, hence their conclusion may not be representative of the population. 73 University of Ghana http://ugspace.ug.edu.gh A chi-square test of association revealed that food type (homemade and kibble) was not significantly associated with helminth infection in dogs. Ahmed et al. (2014), found statistical significance in helminth prevalence among dogs that were fed dry food (kibble) and uncooked food. Zelahem & Mekonnen (2012) also found significant difference in prevalence among dogs that were fed raw food and dogs that were fed cooked and mixed food. 5.3 Intensity of helminth infection in dogs The overall mean faecal egg count of Toxocara spp. was lower than the mean faecal egg count of Ancylostoma caninum in this study. The mean egg count of Toxocara and Ancylostoma caninum in this study was lower than that reported by Komatangi (2005) in Cameroon, Ayinmode et al. (2016) in Nigeria. The mean intensity of A.caninum was higher than the mean intensity reported by Idika & Nwosu (2017) who experimentally infected dogs in Nigeria, Perera et al. (2013) in Sri Lanka, Sowemimo (2017) in Nigeria, de Souza et al. (2015) in Brazil, Degefu et al. (2011) in Ethiopia and Cantó et al. (2011) in Mexico. The difference in intensities may be due to the technique used in estimating the intensities; as the McMaster faecal egg count and Kato-Katz have shown considerable effectiveness in estimating faecal egg count over the formol-ether concentration technqiue (Vadlejch et al., 2011; Rashid et al., 2018; Sowemimo & Asaolu, 2009), geographical variations and management practices (Oliveira-Sequeira et al., 2002). Some studies also estimated intensity at necropsy which is most likely to recover more helminths than coproscopy. S. lupi and D. caninum egg count could not be estimated in this study since they were only observed using formol-ether sedimentation technique and hence reliable egg counts could not be obtained. There was no statistically signifcant difference in mean egg count of A. caninum and Toxocara spp. among breeds in this study. This 74 University of Ghana http://ugspace.ug.edu.gh is contrary to a report by Komatangi (2005) who reported heavier infections of A. caninum and T. canis in local dog breeds. The sex-wise and age-wise mean intensities of A. caninum and Toxocara canis were higher than the intensities reported by Okoye et al. (2011) in Nigeria. In this report and Okoye and colleagues’ report, intensites among sex and age groups were statistically insignificant. This is also in agreement with a report by Degefu et al. (2011) in Ethiopia and Malgor et al. (1996) in Uruguay. However, a report by Katarynza & Błaszkowska (2017) in Poland showed significant association of T. canis intensity with age of dog, where dogs lesser than 12 months old harboured significantly higher numbers of Toxocara canis than dogs above 12 months old. Sowemimo (2007) also reported significant intensities in male and female dogs, with males harbouring more Toxocara canis than female dogs. 5.4 Prevalence of canine haemoparasites A prevalence of 7.69% was reported for Babesia-like haemoprotozoan parasites in dogs. Babesia canis has been reported in dogs in Ghana (Owusu, 2015), so the detection of Babesia-like parasites in dogs in this study was anticipated. Other authors have reported Trypanosoma, Dirofilaria immitis, Hepatozoon canis in Nigeria (Opara et al., 2016) and Ehrlichia canis, Mycoplasma haemocanis, Babesia vogeli in dogs in Australia (Barker et al., 2012). The prevalence and distribution of parasites is known to differ according to geographical location and even climatic variations. The prevalence of Babesia-like parasites was lower than 57.1% that was reported by Opara et al. (2016) in Nigeria. The low prevalence reported in this study is probably due to small sample size or the source of blood. The detection of Babesia has been shown to be feasible in most cases when peripheral blood is taken from the ear or the tail (Jacobson, 2006). In this study, blood was taken from the cephalic vein of dogs and only a few 75 University of Ghana http://ugspace.ug.edu.gh blood samples were collected from the ear. Detection of Babesia in blood film is often difficult in chronic stages than in acute stages of infection (Mosqueda et al., 2012). The diagnosis of Babesia through microscopy is often misdiagnosed as Plasmodium and vice versa because the intraerythrocytic parasites are morphologically identical (Akel & Mobarakai, 2017). However, trophozoites of Babesia can assume tetrad forms and has a characteristic maltese cross, although rare in B. microti, that distinguishes it from Plasmodium (Akel & Mobarakai, 2017). In this study, no tetrad form or maltese cross was seen. However, extracellular merozoites which is characteristic of Babesia (Akel & Mobarakai, 2017) was seen. Infection outcome in dogs can be attributed to ectoparasitic control. 64% of the respondents reported that they did not control ectoparasites on their dogs. Those who reported that they controlled ectoparasite either used acaricides or handpicked the ticks. Picking of ticks by hand has been reported to facilitate infection when it is improperly done (Huygelen et al., 2017). Molecular technique such as PCR is effective and sensitive in the diagnosis of Babesia infection. However, PCR could not be done at the time of the study. Further research involving haematological studies in conjunction with PCR-based detection of haemoparasites in dogs in Ghana is suggested to clarify the potential role of canine haemoparasites and their distribution. Inferences from statistical analysis were limited by the sample number that was obtained, hence inherent risk factors of infection such as age, sex and breed could not be assessed. An insufficient number of exotic dog breeds, puppies and juveniles were sampled for haematological analysis. Local dogs were readily available than exotic dogs and adult dogs were more accessible than puppies and juveniles, hence any inference drawn from their association with infection may not be statistically reliable. 76 University of Ghana http://ugspace.ug.edu.gh 5.5 Management practices, zoonotic implications and awareness on canine zoonoses Management practices such as feeding dogs uncooked offal, irregular deworming and allowing unrestricted roaming of dogs expose dogs to infections. The percentage of owners who do not deworm their dogs (69.6%) is alarming and poses a significant public health risk. Anthelminthic treatment of dogs is effective in controlling the helminth burden in dogs. Feeding dogs uncooked offal exposes them to infective stages of meat-borne helminths such as Echinococcus even after successful deworming. Free-roaming dogs are exposed to parasitic infections as they interact with other dogs that may harbor these parasites. Free roaming dogs may also acquire infections from their coprophagous feeding habits. Hookworms (Ancylostoma caninum) and Dipylidium caninum are common parasites of dogs and are capable of causing larva migrans syndromes and Dypilidiasis respectively in humans (McCarthy & Moore, 2000). Toxocara canis also has zoonotic implications and has been reported in children in Ghana (Kyei et al., 2015). There is a potential transfer of Toxocara through direct contact with the fur of an infected dog (Aydenizöz-Özkayhan et al., 2008). Other reports do not consider this to be a significant risk because the eggs need several weeks to become infective (Overgaauw et al., 2009). One of the objectives of this study was to ascertain owner’s knowledge on canine zoonoses. A greater percentage of the respondents (86.7%) appeared to be well-informed of the potential for rabies transmission from dogs to humans. However, more common and less severe helminthozoonoses and protozoonoses were not known by respondents (0%). Awareness of zoonotic disease risk is a prerequisite for effective prevention (Stull et al., 2012). The limited public’s knowledge of helminth related zoonotic diseases is a public health concern. This clearly reflects the gaps in veterinarian-client education. It is evident that majority of veterinarians do not 77 University of Ghana http://ugspace.ug.edu.gh educate their clients on canine helminthozoonoses and protozoonoses. On the contrary, progress has been made in educating the public on rabies; which is a more serious viral zoonosis. 5.6 Prevalence of equine helminths The overall prevalence of helminths in horses was 39.7%. The prevalence of helminths in this study is lower than the prevalence of 63.7% reported by Tesfu et al. (2014) in Ethiopia, 76.1% reported by Wosu & Udobi (2014) in Nigeria and higher than the prevalence of 15.7 reported by Sawsan et al. (2008) in Sudan. The helminths reported in this study are Parascaris, Strongyle- type eggs and Trichostrongylus sp. Identification of Parascaris to the species level was difficult with microscopy as Parascaris equorum and Parascaris univalens are morphologically identical. Nielsen et al. (2014) reported that Parascaris univalens is most likely to be wrongly identified as Parascaris equorum. Eggs of Strongylus edentatus, Strongylus asini, Strongylus equinus and Strongylus vulgaris are morphologically identical with slight variations in the size of egg (Mehlhorn, 2015). To avoid misidentification, all Strongylus eggs were described as Strongyle- type eggs in this study. Some of these parasites have been reported by other researchers (Umar et al., 2013, Rosa et al., 2018). Several studies have reported significant difference in infection among age groups of horses. In this study there was no statistically significant association helminth infection with age, sex and breed of horses. This is probably due to the small sample size that was used in this study. Umar et al. (2013), however, found significantly high prevalence in stallions and foals used for cadet training in Nigeria. Similar findings have been reported in horses in a report by Regassa & Yimer (2013). Mezgebu et al. (2013) and Fikru et al. (2005) also reported no statistically significant difference in helminth infection among sex in horses. 78 University of Ghana http://ugspace.ug.edu.gh Parascaris was highly prevalent than Strongyle-type eggs in this study. This is in contrast to Mezgebu et al. (2013), Tesfu et al. (2014) and Wannas et al. (2012) who reported Strongyle- type eggs as the most prevalent in their studies. The prevalence of Parascaris (32.9%) in this study is higher than the prevalence of 11.7% recorded by Tolossa & Ashensfi (2013), 4.6% reported by Tesfu et al. (2014) and quite similar to the prevalence of 26.2% reported by Tesfu et al. (2014). Parascaris infection was not significantly associated with age in this study. This is contrary to a report by Hinney et al. (2011) who reported that the true prevalence of Parascaris is age dependent. The prevalence of Strongyle-type eggs (26%) reported in this study is in contrast with the prevalence of 68.8% reported by Umar et al. (2013) in Nigeria. The prevalence of Trichostrongylus (8.2%) in this study was slightly higher than the prevalence of 4.44% reported by Umar et al. (2013). The difference in prevalence can be attributed to management practices, climatic differences, sample size and even diagnostic techniques (Rosa et al., 2018). 5.7 Intensity of equine helminths The mean egg intensity of helminth among age and sex group was highly significant in a report by Tesfu et al. (2014). Contrary to Tesfu et al. (2014), the mean intensity of Toxocara infection among age, breed and sex was not significant in this study. However, the mean intensity of Strongyle-type eggs and Trichostrongylus was significant among age and breed of horses but not sex. The highest egg count of Strongyle-type eggs was observed in local horse breeds and horses more than 10 years old. This is contrary to a report by Tesfu et al. (2014) who reported the highest intensity in horses less than four years old in Ethiopia. A significantly high intensity of Trichostrongylus was observed in horses less than 4 years old and in local horse breeds. This is probably because younger horses do not have strong immune system as adult horses and local 79 University of Ghana http://ugspace.ug.edu.gh horse breeds are not given proper veterinary attention as exotic horse breeds that are valued because they are expensive. 5.8 Prevalence of haemoparasites in horses A prevalence of 10.96% of haemoprotozoan Babesia-like parasites was reported in horses in this study. This prevalence was lower than the prevalence of 22% reported by Oladipo et al. (2015) in Nigeria and 55.0% for Babesia equi, reclassified as Theileria equi in Nigeria (Pam et al., 2013). Infection of horses with Babesia-like haemoparasite may be attributable to interspecies transmission. In the Ayigbe town stable, horses cohabited with domestic animals like dogs, cats and livestock such as cattle. Horses were not kept in stalls and hence were more exposed to infection from infected dogs and cattle that had free maneuverability. It is possible that these animals acquired tick infection elsewhere and introduced them into the stable. Notwithstanding, horses were not treated against ectoparasitic infections at Ayigbe town stable. Horses from Polo grounds stable were routinely treated against ectoparasite by a licensed veterinarian. There was no evidence of cohabitation with other free-roaming domestic animals. This is probably the reason why horses from the Polo grounds stable had no tick-borne haemoparasites. Statistical inferences were not drawn from results due to small sample size. Risk factors such as age, sex and breed could not be assessed with respect to infection because there was great disparity in sample numbers. All horses sampled for haematological examination were local adult breeds. 5.9 Management practices, zoonotic implications and awareness on equine zoonoses Horse management practices such as deworming status, frequency of deworming and food type were same for all horses in both stables. Horses in the study were allowed to sometimes graze in 80 University of Ghana http://ugspace.ug.edu.gh the open field and were given cut grass to supplement their feed. Re-infection in horses is likely to result from their grazing habit or from cut grass (pasture) that is infected with infective stages of a parasitic helminth even after anthelminthic regime. The highly significant difference in helminth prevalence among Polo ground stable and Ayigbe town stable could be associated with management practices. In the Polo grounds horses were dewormed and treated by licensed veterinary personnel. However, horses from the Ayigbe town stable were dewormed and treated by the stable keepers who had no veterinary training. Stable keepers from Ayigbe town stable bought drugs and administered them to horses when they fell sick. It suspected that horses from the Ayigbe town stable were probably underdosed with anthelminthic drugs. Horses from Ayibe town stable cohabited with domestic animals such as cats and dogs and livestock such as cattle. It is possible that some infections may be transferred from these animals to the horses. According to Griffiths (1978), both horses and cattle can host Trichostrongylus; which was only found in horses from Ayigbe town stable. Trichostrongylus axei, a common helminth in horses and cattle has been identified and confirmed by molecular evidence in human infections (Issarapong et al., 2013). Rosa et al. (2018) reported that stables that raised horses together with cattle had an average median intensity greater than others that did not raise horses together with cattle. In all the stables, stable keepers alleged that they cleaned the stalls daily. Cleaning of faeces from stables prevents reinfection of treated horses. None of the stables treated pasture before feeding them to horses. Studies have reported management of pasture is vital to the control of helminths in the stable, hence reducing the rate of anthelmintic treatments in horses (Earle et al., 2002; O'Meara & Mulcahy, 2002). The effect of management practices on helminth infection could not be assessed because all horses in this study received the same husbandry practices except for slight variations in the way management practices were done. None of the 81 University of Ghana http://ugspace.ug.edu.gh stable keepers were aware of any equine zoonotic infection, hence did not exercise sanitary precautions when dealing with horses. This is probably due to lack of education as most of the stable keepers had no formal education. 6.0 CONCLUSIONS, RECOMMENDATIONS AND LIMITATIONS 6.1 Conclusions The study revealed that sex of dog owner, religion of dog owner, reason for keeping dog, educational background of dog owner and occupation of dog owner influence dog ownership. This study determined the prevalence of helminths and haemoparasites in both dogs and horses in Ghana and demonstrated similar prevalence of canine helminths as previously reported by Johnson et al., (2015) in Ghana. Some of the helminths identified in dogs in this study were of zoonotic importance. Toxocara canis was the most prevalent helminthozoonotic parasitic recorded in this study. Dipylidium caninum and Ancylostoma caninum were the other helminthozoonotic parasites reported in this study. A low prevalence of canine Babesia-like haemoparasites was reported in this study. Most owners (69.4%) did not deworm their dogs and owners were predisposed to giving proper care and management to exotic dog breeds than local dogs. Infrequent or non-deworming of dogs was found to be significant risk factors for intestinal helminthiasis. Age, sex, housing unit, food type and manoeuvrability of dog were not significantly associated with infection in dogs. Local dog breeds (Mongrels) were managed under poor management practices as compared to exotic breeds. Significantly more local dogs were managed under free range system, homemade food, and no “veterinary care and anthelminthic treatment”. Majority of dog owners (61%) fed their dogs homemade food and 24.5% of owners fed their dogs off the 82 University of Ghana http://ugspace.ug.edu.gh ground. Majority of dogs (89.7%) were kept for security reasons and 57.7% were unrestricted and roamed with or without supervision. No exotic dog breed roamed without supervision. No dog owner knew about canine zoonotic helminths and zoonotic protozoan parasites. Majority of the owners (86.7%), however knew about canine rabies, yet most dog owners who owned local dogs did not vaccinate their dogs against rabies. Dog owners who kept exotic breeds were found to have good knowledge on dog management and positive attitudes toward dogs than those who kept local dogs. Horses from the Ayigbe town stable were significantly infected with helminths than horses from the Polo ground stable. Horses from the Polo ground stable received proper veterinary care than horses from the Ayigbe town stable. Parascaris was the most prevalent helminth in horses, followed by Strongyle-type helminth and then Trichostrongylus. Age, sex and breed of horses were not significantly associated with helminth infection in horses. Babesia-like parasites were identified in horses at low prevalence. Stable keepers were completely unaware of possible equine zoonoses and hence did not exercise proper precautionary sanitary practices when handling horses or their faeces. 6.2 Recommendations 1. Interaction of multiple variables that affect dog ownership should be investigated. 2. The results show the need for education of dog owners and stable keepers, routine surveillance programmes and preventive health measures such as vaccination and parasite control strategies to mitigate the effect of canine and equine parasitism. 83 University of Ghana http://ugspace.ug.edu.gh 3. It is also recommended that further research on canine and equine haemoparasites and helminths should employ molecular techniques to determine their molecular prevalence and identification to the species level. 4. A wider study that covers several other geographic regions will provide more information on the prevalence and parasite distribution in companion animals. 6.3 Limitations 1. The absence of molecular work in this study made parasite identification based on morphometrics and identification keys quite challenging. 2. The number of horses sampled for coprological and haematological analysis was too small and lesser than estimated sample size. 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Onderstepoort Journal of Veterinary Research, 64, 51-56. 115 University of Ghana http://ugspace.ug.edu.gh APPENDIX APPENDIX 1 : QUESTIONNAIRE FOR THE STUDY OF PET OWNERSHIP PARASITIC INFECTIONS IN COMPANION ANIMALS IN SELECTED COMMUNITIES IN SOUTHERN GHANA Dear Respondent, This questionnaire is designed for an M. Phil research from the Department of Animal Biology and Conservation Science, University of Ghana, Legon. Please FILL IN correct information. All information will be treated confidentially and the information will be used for this research work only. Thank you. Please tick [√] or fill where appropriate. Basic Data 1. Pet Type of animal A. Dog [ ] C. Horse [ ] Breed……………………………………………………………… Age of dog: 0-6 months [ ]1 7-12 months [ ]2 12 months and above [ ]3 Age of horse: 0-4 years [ ]1 4-10 years [ ]2 10 years and above [ ]3 Sex: Male [ ]1 B. Female [ ]2 Sample No……................. Socio-demographic Factors 2. Owner/Caretaker's educational qualification? A. Primary [ ]1 B. JHS [ ]2 C.SHS [ ]3 D. Tertiary [ ]4 Others [Specify]………….....................................…. 3. Owner/Caretaker's Occupation? A. Farming [ ]1 B. Fishing [ ]2 C. Trading [ ]3 D. Office work [ ]4 Others [ Specify] ……………………………………. 4. Religious denomination? A. Christian [ ] B. Muslim [ ] C. Traditionalist D. Other [Specify]……………………………… 5. Sex Male[ ]1 B. Female[ ]2 6. Age. Specify.......................... Nutrition/ Feeding habit of Pet 7. What type of food does dog eat? A. Dog kibble [ ]1 B. Raw fish/meat/offal[ ]2 C. Cooked fish/meat/offal [ ]2 D. Leftover household food3 Other[Specify]................................................ 116 University of Ghana http://ugspace.ug.edu.gh 8. How often do you feed dog in a day? A. Once [ ] B. Twice [ ] C. Three times [ ] Housing, hygiene and health 9. Pet housing A. Kennel [ ] Other [Specify].................................................................. 10. How often do you clean pet housing unit? A. Once daily [ ]1 B. Once every week [ ]2 C. Once every month [ ]3 Other [Specify]....................................................................... 11. How often do you bath pet? A. Once a month1 B. At least twice a year [ ]2 C. Never [ ]3 Other [Specify].................................................................................................................................... 12. On what do you dish out your pet's food? A. Pet bowl [ ]1 B. On the floor [ ]2. Other [Specify].................................................................................................................................... 13. Do you wash your dog's bowl A. Yes [ ] B. No [ ] 14. If No, why? [Specify]........................................................................................................................ 15. How often do you deworm your pet? A. Once every 3 months [ ]1 B. Once every 6 months [ ]2 Other [Specify].................................................................................................................................... 16. Do you take your pet to a veterinary clinic? A. Yes [ ]1 B. No [ ]2 If No, why?........................................................................................................................................... 17. How often do you take your pet to the vet? A. Once every 3 months [ ]1 B. Once every 6 months []2 C. Once a year [ ]3 [Specify]........................................................................................... 18. Where does your pet defecate? A. In the house [ ]1 B. Outside the house [ ]2 C. Inside kennel/housing unit [ ]3 Other [Specify].............................................................................................. 19. How do you control ectoparasites on dog/horse? A. Use acaricides [ ]1 B. Handpick them [ ]2 Other [Specify]..................................................................... Mode of housing 20. What is the housing mode of dog? A. Strictly domestic [ ]1 B. Semi-domestic [ ]2 Purpose of keeping pet 21.What role does pet play in your house? A. Hunting [ ]1 B. Companionship [ ]2 C. Security [ ]3 Owner's knowledge on pet zoonoses 22. Do you know any disease(s) you can contract from your pet? A. Yes [ ]1 B. No [ ]2 23. If Yes, specify............................................................................................................................... 24. Where did you get information on diseases (s) from?................................................................... 117 University of Ghana http://ugspace.ug.edu.gh Presence of Domestic Animals 25. Presence of other animals in household? Yes [ ]1 No [ ]2 Specify type of animal A. Dog [ ]1 B. Cat [ ]2 C. Goat [ ]3 D. Sheep [ ]4 E. Fowl [ ]5 118 University of Ghana http://ugspace.ug.edu.gh 119