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Characterization of Cattle Genetic Resources of KPK through Genetic Markers & Molecular Tech:

NAME OF THE PROJECT:

 Characterization of Cattle Genetic Resources of Khyber Pakhtunkhwa through Genetic Markers & Molecular Techniques.

 LOCATION OF PROJECT:    

Livestock Research & Developmental Station Surezai, Peshawar with the following GPS coordinates.



Format

           Latitude

Longitude

(h ddd.ddddd°)

        N 33.92556°

E 71.65961°




Livestock Research and Development Station, Maira Surizai Payan, Khyber Pakhtunkhwa

PROJECT OBJECTIVES AND ITS RELATIONSHIP WITH THE SECTOR OBJECTIVES:

 

Objectives of the Sector:

The project is directly related to the mandate of the Directorate to conserve and genetically improve the local breeds of livestock through selective and cross breeding. Working under the umbrella of Directorate of Livestock Research & Development and keeping the mandate of the Directorate General (Research) of L&DD department, it demands for achieving the objectives of the project in its true sense.

Objectives of the Livestock Sector (MTDF):

 

1.           Enhance productivity of existing livestock resources through development of new technologies, scientific methods of farming and improved management practices. The new technologies/packages developed regarding different aspects of livestock production in the institute will be communicated to the farmers through print & electronic media, literature, training and extension workers and through other techniques (Inline with the project).

2.             Increase milk and meat and other animal products, by-products and thereby increasing the income of the livestock farmers (Inline with the project).

3.     Exploit export potential of livestock, dairy, poultry and their product/by-products. This will be made through development of new technologies/packages and its communication/transmission to the farmers through media, literature, training and extension network, which will help in increasing the livestock and poultry products and as such will indirectly boost up the export potential of the country (Inline with the project).

4.             Strengthen institutions for livestock research and development and improve their linkages and coordination (Inline with the project).

5.             Promote milk and meat production and its marketing (Inline with the project).

 

Project Specific Objectives:

 

I.              Genetic evaluation of cattle breeds of KP through FAO recommended Simple Sequence Repeats (SSR) markers and molecular screening for gene(s) of interest.

II.             Creation of facilities and introduction of Multiple Ovulation & Embryo Transfer (MOET) technology

III.            Capacity building of the technical staff regarding advanced techniques

IV.           Availability of facility of advanced research in animal genomics and embryo transfer for researchers and academia

V.            Awareness of farmers through trainings regarding breed improvement

 

EXPECTED RESULTS:

 

 

I.               Animals having quality traits identified through molecular tools among the animals sampled will help in the genetic selection of animals at an early age, rather than phenotypic selection for breeding plans.

II.             The techniques introduced will be applied for robust, accurate and detailed evaluation of breeds for better future planning.

III.             Introduction of Embryo transfer technology will enhance the multiplication of elite animals in exponential manner as compared to the linear traditional methods of reproduction.

IV.           The breeds having quality traits will be the source of more income for farmers and will lead to enhancement of socio-economic status of the peoples.

V.            This project will help substantially in the prospective breed development program(s) of the country.

 

DESCRIPTION AND JUSTIFICATION OF THE PROJECT:

Describe the project and indicate the existing facilities in the area and justify the establishment of the project.

 Background:

Pakistan is an agriculture country; livestock is one of the important part of agriculture. More than 70% are related with livestock. It is estimated that there are about 191.3 million of animals (44.4 million buffaloes, 37.7 million cows, 30.1 million sheep, 72.2 million goats, 1.1 million camels).  This livestock wealth is a constant source of milk, meat, eggs, wool, hides and draught contributing substantially to the national economy. More importantly, it is the source of income of the local people especially womenfolk. Majority of the population (90%) live a rural life and each family keep livestock and poultry for their subsistence and meeting other requirements of life. Commercial farming is also gradually gaining weight as law and order situation are restoring to normal. Livestock sector is thus possessing enormous potential and this commodity can be exploited to raise the living standards of the local people. Strengthening the existing veterinary research & development facilities/ institutions focused at solving farmers’ problems will increase animal production and job opportunities.

Table 1: Estimated Livestock Population (Million Nos.)

Species

2014-15

2015-16

2016-17

Cattle

41.2

42.8

44.4

Buffalo

35.6

36.6

37.7

Sheep

29.4

29.8

30.1

Goat

68.4

70.3

72.2

Camels

1.0

1.0

1.1

Horses

0.4

0.4

0.4

Asses

5.0

5.1

5.2

Mules

0.2

0.2

0.2

Source: Ministry of National Food Security & Research (2016-17)

The population of Pakistan is growing day by day at a very faster rate as compare to the developed countries. Simultaneously the number of animals is also growing. Due to this parallel increase in the number of animals and human beings there is increasing competition between them for the available resources like land, water etc.  The developed world has tagelled the situation very cleverly. They are focusing on the increase in the production per animal rather than increase in the number of animals. Developed world has doubled the milk yield per cow in the last 40 years through application of modern tools and techniques for the selection and replication of the producing animals.

 

        Justification:

 The key behind the success in increasing the milk and meat production to a level high enough to meet the needs of the local consumers as well as surplus for export the modern world is the genetic selection for increased production through exploitation of the maximum potential. Animals have been undergoing human-managed selection ever since their original domestication. Initially, selection was probably limited to docility and manageability, but in the last 60 years breeding programs have focused on the genetic improvement of production traits, such as milk yield, growth rate and number of eggs. From the beginning, selection was based on animals’ phenotype with the hope that their offspring would also exhibit superior phenotype. A major advance in selection practice occurred in the mid-20th century, with the advent of quantitative genetics based on principles of heredity and modern statistical theory. In practice, the phenotype of an individual and a substantial number of its relatives is recorded to compute the likelihood that the individual is transmitting a favorable set of alleles for the trait of interest. Although the method is still based on phenotypic selection, it more easily identifies variation at loci having a relatively small effect and represents an important advance. Essentially, the statistical genetics method calculates an average of all genetic loci contributing to a trait as transmitted by the individual, and reports it as an estimated breeding value (EBV). Thus, the animal production industry has undergone dramatic change during the last century. In response to changes in dietary preference of consumers, the selection emphasis has changed from just increased output to increased production efficiency and product quality. In the early 19th century dairy production focus shifted from milk quantity, which was very successfully increased by intense selection, to milk quality and quantity.

Keeping in view the above scenario, this department has started work on various aspects of livestock research and development and cross breeding program of Achai cattle and Jersey

cattle for development of a new dairy breed at Livestock Research & Development Station, Surezai is one such well-planned example.  However, keeping at front the real status of the information available about the breeds of Pakistan at general, and Khyber Pakhtunkhwa in particular, as very little work has been done so far, it is unavoidably needed that the genetic resources of the province should be screened thoroughly at gene level for Quantitative Trait Loci (QTLs) and the information gathered therein shall be used in an effective way, for future breeding plans towards milk and meat production.   

 

PROJECT DESCRIPTION

This project has been designed with three main components.

Genomic Studies

The first part of this project will focus on the genetic characterization of the local animal resources. This study will be conducted using the available machinery and equipment at Genomic Laboratory, Centre of Animal Biotechnology, Veterinary Research Institute, Peshawar. However, some additional machinery and equipment required for such studies will be purchased in this project. For the study purpose, the home tracts of the indigenous cattle breeds, native to Khyber Pakhtunkhwa will be visited to identify true specimen of individual breeds. Breeds of Cattles include: Achai/Gabrali cattle from Malakand division, Lohani cattle from southern region, and Cross breed cattle from Peshawar, Mardan, Charsadda, Swabi and Hazara division. After identification, preliminary data regarding production and reproduction performance at herd/cluster level and at individual animals will be collected to identify the high producing and low producing animals of each breed. Afterwards, individual selected and categorized animals will be sampled for blood for DNA extraction and milk for   total RNA extraction and shifted to laboratory using standard protocols. The samples will be processed using advanced molecular techniques to identify genes of interest and genes expression will be analyzed and estimated using various softwares. The results will be compared between the animals expressing different phenotypes using specie-specific FAO recommended primers (Annexure-VI; Page 34-35). Moreover, for further advanced studies of the same sort (gene mapping and gene targeting) which are highly expensive as very few institutes across country offer such facility, technical and practical help will be sought from experts of other institutions like NIBGE, Faisalabad and NARC, Islamabad and the expenses shall be met from the budget of this project. The data thus obtained will be used as a baseline study and extended to the rest of Animals Genetic Resources (AnGR) of the Khyber Pakhtunkhwa province in future project for better exploitation of the genetic worth and animal resources. No such study has so far been attempted in Khyber Pakhtunkhwa.

 

       Introduction of Embryo Transfer Technology Facility at LR & DS, Surezai

The second portion of this project will focus on introduction of techniques for propagation and replication of elite animals having genes of interest in an exponential way through modern techniques. For this purpose, a Multiple Ovulation and Embryo Transfer (MOET) Technology Center will be established at Livestock research and Development Station, Surezai. At LR & DS, Surezai, as a result of an ongoing cross breeding program of Achai cattle and Jersey cattle breeds since 2012, progeny of first and second generation has been received and for MO&ET studies, these animals will be used. No such study has so far been attempted in Khyber Pakhtunkhwa. The MO&ET Center at LR & DS, Surezai will comprise of an isolated animal shed and MO&ET laboratory and admin block (the details are enclosed in the relevant portion of the DCE attached; Page 40-50 & 52).

   

Other Civil work and solarization work at LR & DS, Surezai

 Fencing work on Southern border of LR & DS, Surezai

 LR & DS, Surezai is situated in Mera Surezai with a total land of 178.2 acres. About 18 acres of land on southern border of the Station is open and to protect the assets and land of the Station from encroachments, the southern border of the Station is very necessary. For such unavoidable reasons budget has been allocated to fencing work on southern border (3008 Rft) of the Station in this project (details are enclosed in the relevant portion of the DCE attached; Page 51).

 

Solarization of tube well at Fodder Section, LR & DS, Surezai

Fodder Section is a sub-station of LR & DS, Surezai, where the research and experiments on development of varieties of fodder and forages varieties is carried out. Moreover, regular staff members of the Station are also using residential quarters located the vicinity of Fodder Section Due to location at a remote area of district Peshawar, the situation of electric power supply is very uncertain and the single tube well is unable to meet the drinking as well as irrigation water needs of the experimental plots. As an alternative, solarization at a water bore in the Fodder Section will overcome the water scarcity for irrigation and drinking purpose, for which, Budget allocation has been made (Details attached at Page No. 57-58).

 

Provide technical parameters and discuss technology aspect of the Project.

This project will comprise of the following three main technical aspects.

 

Molecular Characterization of cattle breeds

This project will focus on cattle breeds of Khyber Pakhtunkhwa province. The other species like buffaloes, sheep and goats will be dealt in future projects.

The home tract of each breed present throughout Khyber Pakhtunkhwa province will be visited for measurements of morphometric parameters (Annexure-VIII, Page No. 37) and sampling will be done from the true specimen of each breed. A proforma will be designed for morphometric measurements of each breed of cattle and possibly buffalo. After thorough screening, both high and low producers (10 animals of each category) will be selected from each breed. Firstly, the milk yield will be recorded based on history, then the milk yield will be calculated using standard bowls. Morphometric measurements will be taken from the animals under consideration and recorded on predesigned proforma having essential informations. Similarly, morphometric observations from the male population and breeding bulls of each breed will also be recorded. 

Both milk and blood samples will be collected from the animals of each category within the breed following standard protocol supported by relevant literature. The samples will be transported in ice pack to the genomic laboratory, Veterinary Research Institute, Peshawar. The samples will be processed for isolation of genomic DNA and total RNA. Blood samples from the male animals and breeding bulls of each breed will also be collected.

 

Molecular work/Genomics

Isolation of genomic DNA

Genomic DNA will be isolated from whole blood samples using DNA extraction kit per the manufacturer’s protocol.

 

Polymerase Chain Reaction

PCR will be performed using standard protocol.

Data analysis:

The data thus obtained will be analyzed using appropriate models, compiled and published.

 

EMBRYO TRANSFER TECHNOLOGY

The demand for livestock and their products is increasing paralleled with the increasing human population. Competition with other uses for land and water resources will also intensify necessitating more efficient livestock production. Application of new technologies inducing genomic selection and reproductive technologies like MOET (Multiple ovulation and Embryo transfer) will play an important role in meeting these demands. Embryo transfer program will include the following steps:

SELECTION OF DONORS FOR EMBRYOS COLLECTION

Animals from the existing pool of already existing breed improvement program will be selected. These animals will be employed for embryo collection.

 

SUPEROVULATION OF DONORS

Reproductive organs of elite donor cows and heifers will be rectally evaluated for different anomalies and cyclicity. Those donors with normal reproductive tract and the presence of corpus luteum (C.L) on one of their ovaries will be selected as potential donors. Donors with C.L will be fitted with a CIDR (controlled internal drug releasing device) for 7 days and will be subjected to superovulation schedule.

Before insertion of CIDR intravaginally, animal will be secured in headlock or chute, first.  Ear tag of the donor scheduled for superovulation will be checked to make sure the animal is correctly identified.

There are three different types of methods currently in use for the recovery of embryos/ova from donors, namely surgical, horn and body flush. Every method has its specific use, advantages and disadvantages. Body flush is easy and there is no need to transfer catheter in between horns.

Embryos/ova from super ovulated cows/heifers will be collected on day 7 following the day the donors were inseminated. Before placing the foley’s catheter in position for flushing, epidural anesthesia will be given taking all the aseptic precautions.

The recovered flushing medium from each horn will be filtered. About 10 cc of the sediment will be poured into searching dish (100x100 mm).

 

CAPITAL COST ESTIMATES

·         Indicate date of estimation of project cost estimates:  July, 2017

·         Basis of determining the capital cost be provided.  It includes market survey, schedule rates, estimation based on previous work done etc.

The capital cost was determined using MRS, 2017 and market prevailing rates.

 

PROJECT BENEFITS AND ANALYSIS:

     •   Financial Benefit:

This is a social service scheme, which aims at increasing the income of farmers through genetic selection of the animals and subsequent dissemination of such animals through MOET technology to the farmers of the entire Khyber Pakhtunkhwa province. These animals will be the source of increasing the income of farmers of the province. With the implementation of the project, facilities of in-time selection of quality animals and culling of unwanted animals will be made available to livestock farmers, saving their money and time. The project proposal has enough potential for disseminating direct and indirect benefits to the farmer’s community. The overall indirect benefits of the scheme would be far more than the expenditure on the implementation of this project.

·               Social Benefit:

·         The project will increase the number of quality animals over the animals of poor genetic quality

·         Reduction in the number of unwanted animals will result in saving of natural resources

·         Improvement in local farming through scientific research

·         Motivation of the people towards farming of low input with high output

·         Research facilities for the students, scholars and scientists will be established

·         Creation of awareness among the stakeholders through publication of research papers, scientific articles, seminars, workshops and trainings.

  • Environmental Benefit:

The project impact will be positive on the environment by earlier genetic selection and dissemination of quality animals. Thorough genetic evaluation of animals for selecting disease resistance animals will reduce the use of unwanted drugs and chemicals etc, for the treatment of animals in the province thus making animals products safer for human consumption. With increase in the number of quality animal and culling of unwanted animals will reduce the waste produced from animals. The sustainability of different ecosystems will be achievable and equilibrium will be assured between ecosystems. Moreover, with the reduction of unwanted animals the natural resources will be conserved.

 

FINANCIAL/ECONOMIC ANALYSIS (WITH ASSUMPTIONS)

FINANCIAL ANALYSIS 

Quantifiable output of the project

With the successful implementation of the project, it is estimated that at least 600 samples will be collected from animals and processed in laboratory for isolation of genomic DNA and RNA and used subsequently in the genomic & genetic studies. This project will be used for creation of animal genomics laboratory and MOET technology facility wherein facility for cryopreservation of quality embryos will be available. At least 02 farmer trainings will be organized for awareness and adoption of Genetic selection and Embryo transfer.  Moreover, three (02) trainings will be organized for researchers and scientists to provide hands on training on MOET. Furthermore, the research conducted will be published in international journals of repute having impact factor.

Employment Analysis

Employment generation (direct & indirect)      

Recruitment in the project will generate direct employment of 16 persons (officers and other staff). Thousands of indirect job opportunities in the area will be produced because of increased livestock/poultry farming.

Implementation of the project:

Indicate starting and completion date of the project      

As soon as project is approved and budget is made available. The project will be completed in three (03) years after the release of funds.

Item-wise/year-wise implementation schedule in line chart correlated with the phasing of physical activities


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