FIRST YEAR PHD SYMPOSIUM ECOLOGY AND EPIDEMIOLOGY OF EHRLICHIA RUMINANTIUM IN NORTH WESTERN NIGERIA FIRST YEAR PHD SYMPOSIUM Student: Babagana Mohammed Adam Supervisors: Prof Richard Birtles Dr Kevin Bown Dr Vincenzo Lorusso

BACKGROUND Ehrlichia ruminantium is the agent of heartwater. MICROBIOLOGY Ehrlichia ruminantium is the agent of heartwater. It is an obligate intracellular bacterium that infects vascular endothelial cells of ruminants.

BACKGROUND TRANSMISSION E. ruminantium is transmitted (trans-stadially) by ticks Most important vector = Amblyomma species

BACKGROUND DISTRIBUTION An estimated number 150,000,000 animals at risk across much of Africa – and infections have recently been diagnosed in the Caribbean

RATIONALE OF THE STUDY Incidence and prevalence not yet documented Increased death rates associated with heartwater Missing epidemiological information HEARTWATER disease. In West Africa, the economic losses resulting from the disease are not yet documented because incidence and prevalence have not yet been researched Efforts to enhance local breeds via crossing with highly productive dairy cattle or susceptible small ruminants from regions that are non-endemic have led to increased death rates associated with heartwater. Presently, there is no safe vaccine. In endemic areas, the controls of heartwater are dependent on acaricides to hinder the transmission, as well as antibiotic therapy missing epidemiological information regarding the diseases in Nigeria. Research focusing on the prevalence and incidence of infections have of late, been hindered by the absence of sensitive and specific tools for diagnosis that are relevant for application in the country. Presently, there is no safe vaccine. In endemic areas, the control of heartwater is dependent on acaricides to hinder the transmission, as well as antibiotic therapy.

AIM To quantify the ecology of E. ruminantium and the epidemiology of heartwater through cross-sectional and longitudinal surveys of livestock, wildlife and ticks in Nigeria

OBJECTIVES To complete a cross-sectional survey of ticks, livestock and selected wildlife to determine the presence/abundance of E. ruminantium and to begin to delineate its transmission pathway. The presence of the organism will be demonstrated using a range of molecular methods including reverse line blotting, semi-nested PCR and real-time PCR, accompanied by comparative sequence analysis of PCR products. To complete a cross-sectional sero-survey of livestock to obtain basic information about exposure rates to E. ruminantium. To complete a longitudinal survey of livestock and tick samples for E. ruminantium to define the seasonal dynamics of (i) tick populations and (ii) E. ruminantium infections in ticks and blood samples of livestock.

STUDY AREA Ecological zone: Guinea savannah Season : Raining (May-Sep) Dry season (Oct-Apr) Land Area : 33,411 Hectares People: 5,252 Cattle: 23,327 Sheep: 5,914 Goats: 5,058

BLOOD SAMPLE COLLECTION 10ml Blood Jugular vein A drop on FTA Card 5ml To EDTA bottle 5ml To plain bottle Haematology analysis Serology analysis Molecular analysis Salford University Laboratory Ahmadu Bello University Laboratory

TICK SAMPLE COLLECTION 70% Ethanol Molecular analysis University of Salford

Agarose gel electrophoresis MOLECULAR ANALYSIS Blood Tick DNA EXTRACTION Chelex Method Ammonium Hydroxide Ehrlichia primers Presence of pathogen was determined using Ehrlichia Agarose gel electrophoresis

RESULT PREVALENCE OF EHRLICHIA RUMINANTIUM IN TICKS Number of ticks = 141 Number of positives = 28 Presence of pathogen was determined using Ehrlichia RESULT Anaplasma platys

RESULTS PREVALENCE AND DISTRIBUTION OF TICKS Genus Frequency Percentage Valid Ambloymma 113 33.8 Rhiphicephalus 22 6.6 Hyalomma 6 1.8 Total 141 42.2 Missing System 193 57.8 334 100 Number of animals = 334 Number of ticks = 141

RESULTS CATTLE: High prevalence in adult 19.70% and low 2% young PREVALENCE AND DISTRIBUTION OF TICK INFESTATION WITH AGE   Species Total Sheep Goat Cattle AGE Young Count 46 50 2 98 % within AGE 46.90% 51.00% 2.00% 100.00% % within Species 31.10% 36.80% 4.20% 29.50% % of Total 13.90% 15.10% 0.60% Adult 102 86 234 43.60% 19.70% 68.90% 63.20% 95.80% 70.50% 30.70% 25.90% CATTLE: High prevalence in adult 19.70% and low 2% young SHEEP: Low prevalence in adult 43.6% and high in young 46.9% GOAT: High prevalence in young 46.90% in and 43.60 in adult

RESULTS PREVALENCE AND DISTRIBUTION OF TICK INFESTATION BASED ON SEX   Sex Total Male Female Genus Ambloymma Count 59 53 112 % within Genus 52.70% 47.30% 100.00% % within Sex 88.10% 72.60% 80.00% % of Total 42.10% 37.90% Rhiphicephalus 5 17 22 22.70% 77.30% 7.50% 23.30% 15.70% 3.60% 12.10% Hyalomma 3 6 50.00% 4.50% 4.10% 4.30% 2.10% 67 73 140 47.90% 52.10% SEX: High tick infestation was recorded in female 52.1% than male 47.9%

CONCLUSION This study revealed that hard ticks of three genera namely; Ambloymma Rhipicephalus Hyalomma were prevalent Soft ticks were not encountered during the study. Young animals were found to have the lowest prevalence than the adult animals group. Furthermore, female animals were more susceptible to tick infestation than their male counterparts and sheep are more infested with ticks compared to goat and cattle.

WORK SCHEDULE Fieldwork between July to September 2016 During the 3-month fieldwork period, the blood samples collected will be centrifuge to separate it from the serum October 2016 to July 2017, the blood and tick samples collected will be analysed