PROPOSAL ON

PREVALENCE OF AVIAN TUBERCuLOSIS AND ASSOCIATED RISK

FACTORS IN SMALL AND LARGE SCALE POULTRY FARMS IN AND
AROUND DIRE DAWA, ETHIOPIA

BY

AMIN ABDO

ADVISOR:

Dr. GEZAHEGNE MAMO

October, 2013

DEBREZEIT, ADDIS ABABA

1. INTRODUCTION

The world poultry population has been estimated to be about 16.2 billion, with 71.6 % in
developing countries, producing 67, 718,544 metric tons of chicken meat and 57,861,747 metric
tons of hen eggs (Gueye, 2005). In Africa, village poultry contributes over 70% of poultry
products and 20% of animal protein intake (Kitalyi, 1998). In East Africa over 80% of human
population live in rural areas and over 75% of these households keep indigenous chickens
and Ethiopia is not exception to this situation (Kitalyi, 1998).

Ethiopia has large population of chicken, estimated to be 42 million (CSA, 2009). Recent
estimates put the poultry population in Ethiopia at around 40.6 million with native
chicken of none descriptive breeds representing 96.6%, hybrid chicken 0.55% and exotic
breeds of chickens mainly kept in urban and peri-urban areas 2.84% (CSA, 2009). From the
total population of chicken in Ethiopia, 99 % are raised under the traditional back yard system of
management, while 1 % is under intensive management system (Tadelle et al., 2003).

Ethiopia is representative of countries where village poultry plays a dominant role in total poultry
production. The sector represents an important part of the national economy in general and the
rural economy in particular. In developing countries, many rural households keep poultry in their
farmyard. The importance of village poultry production in the national economy of developing
countries and its role in improving the nutritional status and incomes of many small farmers and
landless communities has been recognized by various scholars and rural development agencies
for the last few decades (Abera and Tegene, 2011). For instance, there are about 42 million
chickens in Ethiopia of which 96.6% are local chickens (CSA, 2009/10), indicating the
significance of indigenous chickens as potential Farm Animal Genetic Resources of the country.
The impact of village chicken in the national economy of developing countries and its role in
improving the nutritional status, income, food security and livelihood of many smallholders is
significant owing to its low cost of production (Abubakar et al., 2007). Several factors have been
suggested for the low production characteristics of free-range village chickens. The system is
characterized by low input and low output, with minimal management interventions, feed
supplementation, housing and disease control. This kind of production by itself is a limiting
factor to sound economic and sustainable production. The low input might be, however, a result
of the high risk due to high mortalities experienced in village poultry. Thus, the potential of the
free-range chicken production has not been exploited. Therefore, if any success is to be achieved
in improvement for free-ranging chicken production, it will inevitably depend on the successful
control of major poultry diseases (Nassir, 1998; Tadesse et al., 2005).

In Ethiopia, poultry diseases are considered the most important factor responsible for reducing
both the number and productivity of chickens (Abubakar et al., 2007). Avian tuberculosis is one
of the most important diseases that affect domestic and pet birds. Several Mycobacterium species
can be involved in the etiology of avian tuberculosis. The disease is most often caused by
Mycobacterium avium belonging to serotypes 1, 2, 3, and 6 and also M. genavense. Other species,
such as M. intracellulare, M. scrofulaceum, M. fortuitum, M. tuberculosis, and M. bovis can also
cause avian tuberculosis, but the incidences are rare (Fulton RM et al., 2003).
Infected birds (as they shed large amounts of organism into the environment) and contaminated
water and soil (as the Mycobacteria can survive for several months in the environment) are the
main source of infection. The bacilli are extruded from ulcerated lesions of the intestine and are
voided in droppings. The most common route of infection for susceptible birds is the alimentary
tract (Dvorska L et al., 2007). Respiratory tract is also suggested as a potential source of
infection. The disease gets transmitted to the susceptible birds by ingestion and inhalation of
aerosolized infectious organisms. The disease is more prevalent in places with high population
density and poor sanitation and hygienic conditions. The practices of allowing birds to roam
freely and keeping the breeders for several years are highly conducive to the spread of
tuberculosis. Persistence within flocks is associated with keeping older stocks without following
adequate cleanliness and hygiene (Fulton RM et al., 2003). Treatment of infected animals is
rarely attempted because of the high cost and prolonged time. In avian therapeutics related to
mycobacterial infections, the major difficulty is that the pharmacokinetics in birds for most of the
antimycobacterial drugs is unknown (VanDerHeyden N, 1997).

Therefore, the objectives of this study are to:

Determine the prevalence of avian Tuberculosis in poultry farms and its association with
different risk factors in Dire Dawa area.
2. MATERIALS AND METHODS

2.1. Study Area

The study will be conducted in Dire Dawa area, from October 2013 to May 2014 which is located
in the Eastern part of the country bordering by the state of Somalia and the state of Oromia. It is
found at the distance of 515 kilometre from Addis Ababa, lies between 1000 to 2000 metre above
sea level in between 09°28'N to 09°49'N latitude and 41°38' to 42°19'E. The administrative has
area of 146,802 hectares. Diredawa is grouped in kola and semi kola climatic zone. It has bi
modal rainfall pattern with the highest rainfall in July and August. The average annual rainfall
varies from 700mm in dry (kola) to 900mm in weinadaga zone. The rainy season lasts from July
to September. The mean annual tempreture varies from 20 to 30°C in kola and 14 to 20°C in
weinadaga. The hottest month of the year ranges from 28.1°C record in December and January to
34.6°C record in May (DDRAB, 2007).

2.2. Study animals

The study animals are Bovans (exotic breeds) and local breeds of chickens in poultry farms of
Dire Dawa and owned by local farmers in and around the town. The study animals will be
grouped into sex (male and female), breeds (exotic and local) and ages as young (2-8 weeks)
and adult (above 8 weeks of age).
The chickens used in this study will be those reared entirely by local poultry farms. The principle
is that the health status of chickens taken from farms could reasonably reflect the actual health
status of the population from which they are taken.

2.3. Sample Size Determination

The animals will be selected by using simple random sampling method. To determine the sample
size, an expected prevalence of 6.3% will be taken from the previous research worked in central
area by Serkalem Tadesse, veterinary doctoral thesis_2001. The desired sample size for the study
will be calculated using the formula given by Thrusfield, M. (1995).
N=1.962 Pexp (1-Pexp) Where, N= required sample size, Pexp = expected prevalence,
 d2
d = desired absolute precision, 1.962 =z- value for 95% confidence interval
Using this formula, the estimated sample size will be 91 chickens.

2.4. Study Design and Methods

A cross-sectional study will be conduct on avian tuberculosis in randomly selected indigenous
breed chickens, reared under extensive system in and around Dire Dawa despite of their sex, age,
and health status.

2.4.1. Examination procedure and Questionnaire

A total of 91 apparently healthy local chickens were to be purchase & then examined or simply
examined from local open-air markets and on house-to-house visit basis respectively. A
structured questionnaire survey will be perform in parallel with direct observations of flocks to
individuals and households who were directly responsible for the care of chickens both in market
place and during house-to-house visit basis on the study areas. Before undergoing any
examination procedures, each chicken will given by an identification number. Just before
necropsy, ante-mortem examination will made for any abnormalities and clinical signs of any
disease. During ante-mortem examination, physical parameters (live weight, carcass weight,
thigh circumference, and breast dimension) will measured, and clinical signs considered
indicative of avian tuberculosis will recorded before the chickens were killed.

2.4.2. Postmortem and histopathology Examinations

For postmortem examinations, the chickens would be killed and opened according to the
procedures of Zander and Mallison. At necropsy, all internal organs will be examined and any
observed gross lesions would be recorded on necropsy card. Tissues from organs showing
pathologic gross lesions will be collected in 10% buffered formalin. Furthermore, tissues from
liver, spleen, and pieces of intestine at different segments will be taken for histopathology
regardless of the presence or absence of lesions and for the reason that they can be highly
affected by the MAC group. After possible post-mortem examination in the study area, samples
which will be taken from different organs of both tuberculin positive test chickens and
characteristic positive post mortem lesions will be then transport to Aklilu Lemma institute of
pathobiology research laboratory for subsequent and further laboratory examination (culturing
and molecular tests).

2.4.3. Mycobacteriologic Culture

Of the 91 chickens selected randomly in and around Dire Dawa, samples from positive chickens
during ante-mortem examination and tuberculin tests to be cultured on sodium pyruvate-enriched
Lowenstein-Jensen (L-J) media.

2.5. Data Analysis

The prevalence rate of avian mycobacteriosis is defined as the number of cultures that are
positive from the total number of animals examined expressed in percent. Culture is considered
here as the determinant of MAC as it showed the acid-fast bacilli definitively and used as “gold
standard”. Postmortem examinations and special staining in histopathology were also used to
further confirm cases. Altitude, sex, and age dependent prevalence were compared and analyzed
by the chi-square test (FREQ procedures of the Statistical Analysis System).

3. EXPECTED OUTCOME

The study will be undertaken with the objective to study the prevalence of
Avian Tuberculosis in Dire Dawa area. Based on the determinants associated
with Avian Tuberculosis, a clear and feasible recommendation on the control and prevention
measures of the disease, and how to upgrade this poor management of traditionally reared local
chickens will be forwarded.

4. ACTION PLAN
 Octob Novemb Decemb Januar Feburar Marc Apri Ma Jun,20
er, er, 2013 er, 2013 y, y, 2014 h, l, y, 14,
2013 2014 2014 201 201
4 4
Proposal 
submission
Field and 
farm
visiting
Questionnaire  
Tuberculin test  
ing, follow-
up & post-
mortem
examination fo
r screening
Culturing of     
M. avium

Data entry  
and
analysis
Thesis write  
up

Thesis 
submission
and
defense
5. COST BREAK DOWN

Table-1: Cost of material for Tuberculin testing, follow-up and post-mortem examination.
No Item Unit Quantity Price/ unit Total price
1 Glove, gown and mouth
mask
2 Cap and boot 2 each
3 Scissor and cutting knife 4
TOTAL = >1000 ETB
Table-2: Cost during Surveillance and transportation including cost of food, bed and parking

Travels Unit Food &

of Cost/unit Bed or Total cost
travel travel parking
cost/travel
Debire-zeit to Dire Dawa & vice versa 2 300 200 1000
Dire Dawa to AA for sample submission to 2 300 200 1000
Akililu Lemma Institute of pathobiology & vice
versa
House to house & house to market Surveillance - - - ≥500
in different Woredas .
Dire Dawa to different Woredas & vice - - - ≥500
versa including house-to-house & house-to-
market Surveillance.
House –to- house visit in Dire Dawa Town - - - ≥100
House –to- market site visit in Dire Dawa Town - - - ≥100

TOTAL = 3200 ETB

Table-3: Stationary cost

Item Unit Quantity Unit of cost Total cost

Video-camera 1 1 3000 3000
Flash disc 8GB 1 200 200
Pen Dozen 12 42 42
Marker 3 3 10 30
Ruler 2 2 5 10
A4-size paper 1 400 120 120
Photocopy 100
Print 100
CD-RW 2 2 40 80

Internet 4 hr/day 4 day/week=16 25cents/minute=15 8640

service day/month=14 ETB/hr=60ETB/day(4hr)
expense 4 day/ 9 month

Note-Book 1 1 30 50
TOTAL = > 12,372 ETB

• Costs for reagents and materials that used for microbiological culture of M. avium

Reagents: Cost

 Decontaminant = 4% NaoH
 Neutralizer = H2SO4
 Indicator = 1% Bromocresol-purple
 Phosphate buffered saline solution (BPS).
 L-J egg media with sodium pyruvate (Difco laboratories, Detroit, Michigan 48201).

Materials:

 Sterile glass pestle and mortar, and 50 ml screw capped centrifuge tubes.

Total cost = >1000 ETB

TOTAL BUDGET = >1000 ETB + >1000 ETB + 3200 ETB + >12,372 ETB = >16,572 ETB
6. REFERENCE

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Abubakar, M.B., Ambali, A.G., and Tamjdo, T. (2007): Rural Chicken Production: Effects of
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