Moses Danlami Ogah Phd Proposal

CHARACTERIZATION AND GENETIC STUDY OF INDIGENOUS MUSCOVY DUCK FROM TWO AGRO- ECOLOGICAL ZONES OF NIGERIA

OGAH.D M ./PhD/1966/06

Major Supervisor: Professor Dr. N. I. Dim Minor Supervisor 1: Dr M . O Mommoh

CHARACTERIZATION AND GENETIC STUDY OF INDIGENOUS MUSCOVY DUCK FROM TWO AGRO- ECOLOGICAL ZONES OF NIGERIA

1.0

INTRODUCTION

Poultry production is one area of animal production with significant contribution to human food production. Consumers have high preference for poultry products. Beside preference; poultry products provide protein of high biological value (Epstein, 1990). Nigeria is endowed with many poultry species which are local to the country .These have lived, adapted and produced for several years in Nigeria environment(Momoh, 2005) .RIM (1992) estimated poultry population to be about 33 million. With the ever growing population and concomitant improvement in the living standard of Nigerians ,the demand for egg and other poultry products will continue to grow thus overstretching the supply from chicken .Therefore,there is need to explore other poultry species such as the duck to supplement the eggs and other poultry products obtainable from the chickens. The duck is widely distributed and is prominent in the list of available poultry species in Nigeria (RIM, 1992) They are essential part of many human societies around the world. They are reported to be domesticated as early as 2500 BC (Farrel,1995).Today the domestic duck have come to serve a variety of roles in modern society as source of food supply (meat and egg ),and in some other societies as a species that is selectively bred for shows. They have several advantages over other poultry species .These advantages include their disease tolerance , hardiness , excellent foraging ability and easy to herd, particularly in wet lands since they tend to flock together (Farrel,1995).

The population of ducks in Nigeria was put at approximately eleven million(11,000,000) and is reported to be distributed all over the agro ecological zones of the country (FLDPcs,1992).Domestic ducks were ranked third among various poultry species in Nigeria (Hassan and Mohammed, 2003). In spite of this positive score, very little attention has been paid to the genetic improvement of the duck that will lead to increased productivity. There are little or no available information on the phenotypic and genetic characteristics of Nigerian indigenous Muscovy duck. In Nigeria, different agro ecological zones exist; and for all species of livestock variations are reported to exist in both phenotypic and genetic characteristics as a result of adaptations to these different ecological zones. Numerous comparative studies have provided insight into the ecological mechanism underlying evolutionary diversification across habitat gradient .This is evident in the reports of Akinokun (1971), Oluyemi et al (1982),Adedokun and Sonaiya(2001)for poultry; Adebambo et al. (1998) for cattle,Epstain(1990) and Wilson(1991) for sheep and goat. These diversities constitute valuable animal genetic resource for use in animal breeding programme for improvement of the productivity of the animal species, particularly domestic poultry and also in designing proper conservation strategy In genetic studies various type of genetic makers are known, they include morphological, chromosomal, biochemical and molecular makers. Morphological (e.g. pigmentation and other features) and chromosomal (e.g. structural or numerical variations) makers usually show low level of polymorphism or differences. Quite a number of techniques are in use in molecular maker analysis; the most common and cost effective is the Random amplified polymorphic DNA technique. This technique has been used extensively in molecular characterisation of livestock population in detecting polymorphism between population and establishing genetic relationship among them (Ali et al., 2003). Ecotypes are developed through selection and adaptation and varied from one agro ecological zone to another due to varied climatic characteristic such as humidity , temperature and rainfall ,though in some cases some ecotypes may be similar in all traits (Adedokun and Sonaiya ,2001)

Some research findings on ducks Ksiazkiewic (1995) and Farrell(1995) reported that they are very susceptible to effective inbreeding and genetic drift if kept in a small population for few generation .This genetic flexibility will suggest that variation can easily be found in different population at different location .These variation could either be morphological or genetic or both. .The genetic improvement of local ducks in Nigeria will first require investigation into their productivity genetic characteristics and geographical diversity.

1.1

Research Objectives

The overall objectives of this study will be to evaluate the phenotypic and genetic characteristics, as well as the potential of the local Muscovy ducks to contribute to egg and meat production in Nigeria. This information will provide a frame work for the development of breeding programmes for conservation and sustainable use of the Muscovy duck genetic resources in Nigeria. The specific objectives include 1. To evaluate the morphological/phenotypic characteristics of local Muscovy ducks from two agro ecological zones of Nigeria (tropical rainforest and guinea savannah ) 2. To estimate morphological distance between the two populations 3. To evaluate performance of the ducks from these zones in basic production traits, of growth (0-to 35 weeks), and egg production. 4. To estimate genetic parameters of growth, egg production and fertility traits of the ducks from the two ecotypes. 5 To estimate genetic relatedness among the two ecotypes using random molecular techniques .

2.0

Materials and Methods

Two approaches will be adopted for the research, field measurement or field data collection and onstation

experiment;

2.1.0 Field data collection; Characterization study A field measurement to characterise indigenous Muscovy ducks will be carried out in two agro ecological zones. 2.1.2 Study site- The two agro ecological zones will

include Rainforest and Guinea savannah

areas. The rainforest agro ecological zone lies at the southern part of Nigeria and data will be collected from southern part of Cross River State, Akwa Ibom and Abia states. Guinea savannah agro ecological zone – The data will be generated from rural areas of Benue, Nasarawa and Niger states. 2.1.3 Sampling method-participatory rural/visual appraisal and physical measurement will be adopted. The method will involve taking measurement of traits from the two ecotypes 2.1.4 Traits to be measured – 1 Morphological data Body weight Body length Body width

Head length

Beak length

Neck length

Beak width Beak height

body morphology and external egg

Wing length Shank length Bird height

2 Egg Characteristics Egg weight Egg length Egg width Shape index

2.2.0 .Phenotypic evaluation of production traits-On station experiment. On station experiment to evaluate the phenotypic, production and genetic characteristics of the indigenous muscovy ducks from the two agro ecological zones , Rainforest and Guinea savannah will be carried out in the Poultry Unit of College of Agriculture ,Lafia , Nasarawa state of Nigeria. 2.2 .1 Screening of nondescript population of the Muscovy ducks from each of the agro ecological zones to form foundation or base population. Ten adult drakes and seventy ducks (female) will be randomly gathered from rural areas of Southern Cross River, Akwa Ibom and Abia states to make up the nondescript populations from the Rainforest, while similar adult male and females will also be gathered from rural areas of Benue, Nasarawa and Niger states to also form the nondescript population of the birds from Guinea savannah zone. Only matured adult that have laid and can mount and are physically Sound will be collected. 2.2.2

Generation of experimental birds from the base population (random bred population)

The non descript population will be assigned into ten replicate pen per ecotype, in a mating ratio

of 1:7 (male /female) randomly and will be fed layers marsh .They will be allowed to lay eggs and hatched naturally by brooding mothers in batches The ducklings will be brooded in separate pens for eight weeks. At ten weeks of age the breeding groups will be selected to form each ecotype, ten males and seventy females as optimum recommendation by Nickolova (2004) for Muscovy duck. The ducks will be mated at 28 weeks of age.

2,3.0 Genetic evaluation of the Muscovy ducks (straight bred mating) Seventy females and ten males foundation stocks of each ecotype obtain from a random bred Population will be place in a mating ratio of 1:7 (i.e. one male to7female).This is to generate straight bred from first laying ducks of each ecotype. The distribution and mating of each ecotype will be at random. 2.3.1 Traits to be measured The following traits will be measured for the phenotypic and genetic evaluations.

Growth trait - Body weight –each genetic group will be weighed at 5 weeks interval from hatch 35weeks of age - Daily weight gain - Body weights

gain at 5 weeks interval

- Specific growth rate - Mortality from 0-35 weeks of age. Egg production traits - Age at first egg - Body weight at first egg.

to

- Percent egg production - Weight of first egg - Egg weight at 30, 35,40,45,50, weeks - Egg mass - Number of egg laid at 30, 35, 40, 45, 50, week. - clutch size - Laying intensity

Fertility and hatchability traits - Hatchability of fertile egg (%) -Average duckling weight

2.4.0 Molecular characterization Protocol/ procedure -Blood sample of about 500ul will be collected from the brachial vein of 25 individual birds of either sex from each of the two ecotypes. -Genomic DNA will be extracted by the use of phenol chloroform extraction method using the protocol adopted by Hesfer (1997). -The pellet of DNA will be washed and dried. The concentration of the DNA and its purity will be determined by spectrophotometer based on the absorbance at 260 and 280nm respectively. The purified DNA from individual as well as pooled DNA from each ecotype will be used for further analysis.

3.0

ANALYSIS OF DATA

3.1

Phenotypic evaluation

For morphological traits, growth, egg production, fertility,, and short and annual term egg Production, factorial analysis of variance in a complete randomised design using appropriate factors such as ecotype/genotype, sex, hatch, and interaction will be used . A generalised linear model (GLM) procedure will be adopted using Statistical Analysis System (SAS) (1998) Using the following model Yijk l=µ+Ei+Sj+Mjk+ (ES) ij +eijkl Where Yijk=individual animal

µ =overall population mean Ei=the fixed effect of ecotypes/genotype (1,2 ) Sj=effect of sex (1,2) Mjk=Random effect of sire within ecotype (k=1…7) (ES) ij=Effect of Ecotype x Sex interaction

eijkl=Residual error assume NID(0,σ2)

3.2

Morphological distance Morphological data will be use to estimate genetic distance between the ecotypes. Nine zoometric traits will be used. body weight body length body width neck length beak length beak width

beak height shank length head length

Using the means procedures of the SAS (1990) package (Statistical Analysis System Institute Inc 1990) the simple descriptive statistics of each numerical variable will be obtain step wise discriminatory analysis will be perform to assess the discriminatory power of each variable ,genetic distance will be obtain using Mahalanobis D² statistics using the means of each discriminant variables monitor.(Salako and Ngere,2001 ) D² =∑ ∑ Vij (xi-yi)(xj-yj) Where D²=geometric distance between two population represented in an m-dimensional space Vij=is the element of ith row and jth Column of the inverse matrix - The morphological data will also be subjected to principal component analysis (PCA) for multivariate analysis to reveal pattern within the data matrix.

3.3

Genetic and phenotypic parameter estimation Growth, egg production, and fertility data of each ecological group will be Subjected to genetic analysis using the mixed model least square and maximum likelihood Package Harvey (1990) to estimate genetic and phenotypic correlation as well as heritability Estimate of traits from sire variance components.

3.4 Random Amplified Polymorphic DNA-Polymerase Chain Reaction Analysis (RAPD-PCR) RAPD –PCR will be carried out with the pooled and the individual genomic DNA samples .Five random primers will be use and amplified by polymerase chain reaction (PCR).Each sample will be use for electrophoresis.

RAPD pattern will be visualised on a Ultraviolet (UV) transilluminators and photograph. Recording of data and statistical analysis The RAPD bands will be score for their presence (1) or absence (0) .The index for similarity between ecotypes and within ecotype will be calculated using the formular developed by (Lynch, 1990) . Bab= 2Nab/Na+Nb Where Nab= number of fragment observed in individuals a and b Na and Nb = total number of fragment scored in a and b

3.4,1 Genetic distance base on band sharing The genetic distance between the populations will be calculated based on band sharing between the pooled sample profiles. The genetic distance between ecotypes will be calculated as developed by (Chatterjee et al 2007) Dab=1/N.1- (Nab/Na+Nb-Nab) Where Nab= number of common band between ecotypes Na= number of common band in ecotype a Nb=number of common band in ecotype b N= number of primers

4.0

REFERENCES

Adebambo O, J.L.Williams, Sara Blot and Barbara Urquhart (1998) Genetic variation in Nigeria cattle breeds using 26 microsatellite marker .In proceeding of silver jubilee of Annual conference West Africa society for Animal Production .March 24-26pp Adedokun S.A.and Sonaiya E.B.(2001) Comparism of the performance of Nigeria indigenous Chicken from three agro ecological zones Livestock Research for Rural Development 3(2) 34-39 Akinokun O.(1971) The problem of poultry production in Nigeria .Paper presented at the 7th Annual Conference of Agricultural Society of Nigeria. Kano

Ali B.A , Ahmed M.M.M. and Aly OM (2003) Relationship between genetic similarity and some Production traits in local chicken strains. Africa Journal of Biotechnology 2 (2) 46 -47 Barker J SF , Tan S G, Moore SS , Mukhere TK ,Matheron JL ,Selvaraj OS (2001) Genetic variation within and Relationship among population of Asian goats (Capra hircus) Journal of Animal Breeding and Genetics 118 :213 -233 Chatterjee RN, Sharmer RP ,Reddy B LN, Niranja M, Shivaprasad and Mishra S k (2007) Genetic Analysis of highly inbred chicken using RAPD_PCR and Immunocompetence. Intern. Journal of Poultry Sc. 6(12) 967-972

Ehiobu, N G., and Goddard, M. E. (1989) Heterosis in crosses between lines of Drosophila melanogaster selected for adaptation to different environments .Theor. Appl. Genet. 77:253 -259 Ehiobu, N G ., Goddard, M. E. and Taylor, J .F (1990)Heterosis in crosses between geographical Separated populations of Drosophila melanogaster. Theor. Appl. Genet. 80(4):569-575 Epstein H.(1990) The context and prospect for development of smallholder poultry production In Africa. In small holder rural poultry production .proc CTA seminar G13 Thessalonica Greece35-50 Farrell D.J.(1995) Table egg laying ducks:nutritional requirement and current husbandry System in Journal of Avian Biology Review 6(1995)55-69 FDLPCs (1991) Federal Department of Livestock and Pest Control services. Federal Government of Nigeria 287- 288 Girard P.and L. Palabost (1976) Etude du polymorphisme enzymatique de 15 population natureles de drosophila melanogaster Archives de zoologies experimentale et genetics117:41-55 Harvey W.R. (1990) Mixed model least squares and maximum likelihood computer programme .pc2 Mimeographs. Ohio State University press Ohio Hassan W.A and W.S. Mohammed (2003) Ecotypes of the Muscovy in the North West of Nigeria; Variation in body weight and bill length .In proc. of 8th Ann. conf.of Animal Science Association of Nigeria 2003. Ksiazkiewicz J. M (1995) Duck gene pool. International Symposium on conservation measures for rare farm Animal breeds .Balice –Poland (1995)289-292 Hesfer M (1997) Targeted mapping of chicken genome PhD Theses submitted to University of Hertfordshire

Lynch M (1990) The similarity index and DNA fingerprinting Mol. Biol 7:478-484 Momoh O. M (2005) Genetic and Phenotypic Evaluation of the Nigerian Heavy Chicken Ecotype and Its Cross Bred with the Light Ecotype PhD Thesis University of agriculture Makurdi.2005 Nwosu C .C, Gowen F. A, Obioha F.C., Akpan I. A.andOnuora (1985) A biometrical study of the Conformation of the native chicken .Nig. J. Anim .Production 12(1&2) 141-146. Nickolova M. 2003 Study on some major reproduction factors of Muscovy ducks (Cairina moschata with element of the incubation technology PhD thesis Agricultural University of Plovdiv. Bulgaria 2003 Oluyemi J .A.Longe G.O.and Songu T. (1982) Requirement of the Nigerian indigenous fowl for Protein and amino acid .Ife Journal of Agric.4:105-110. RIM (1992) Nigeria Livestock Resource vol.11National Synthesis Annex publication. Resource Inventory Management limited Salako A K and Ngere L.O. (2001) Genetic distance between West African Dwarf (WAD) and Yankasa Sheep in South West Nigeria. Nigerian Journal of Genetics 16(2001) 37-43 Saitbekova N, Gaillard C, Obexer-Ruff G, Dolf G (1990) Genetic diversity in Swiss goat breeds based on microsatelite analysis Animal Genetic 30:36-41. SAS (1990) SAS user guide statistics SAS Inc Cary N C 633. Wilson R. T (1991) Small Ruminant Production and the small ruminant genetic resource in tropical Africa FAO Animal Production and Health Paper no 88 23pp Wilhan JGK, Kubelik AR, Livak KJ, Rafalski JA , Tingey SV (1990) DNA Polymorphism Amplified by arbitrary primers are used as genetic markers .Nucleic acid Research 18:6531-6535

Appendix Body measurements

Body length- measure between the first cervical vertebrae and the pygostye Bird height – measure from the legs on the ground up to the back of the body. Body width – distance between the right to the left flank of the body. Beak length – measure as length of the upper beak rim Beak width – at the widest part of the beak between the right and the left distance Beak height - at highest part of the beak. Shank length – from the knee or knuckle (hock joint)to the region of the tarsus Wing length – measure as the distance from the caput humeri to the third carpal digit Head length –as distance between the end of the beak and the end condylus occipitale Neck length –measure between the first and the last cervical vertebrae Head width – at the widest part of the head Head height- at the highest part of the head.