ANIMALSCIENCES - Pakistan Agricultural Research Council...

ANIMAL SCIENCES

ANIMAL SCIENCES

The Animal Sciences Division (ASD) of PARC is cognizant of the importance of livestock sector innational economyand efforts are underway to tap the potential and opportunity's offered by the sectorfor enhancing national food security, economic uplift, rural development, poverty alleviation,employment generation and foreign exchange earnings . The main research focus of ASD has beencontrol of infectors diseases of livestock and poultry (FMD, PPR, HS, AI, ND), improving feedingmanagement strategies, increasing reproductively efficiency and boosting small ruminantsproductivity. The ASDresearch activities are undertaken mainly at Animal Sciences Institute . NARC.

Research Activities

ANIMAL HEALTH

ANIMAL SCIENCES

Livestock industry plays an important role in the economy of Pakistan . However, prevalence ofinfectious diseases poses a major threat to the livestock productivity. The research focus of the AnimalHealth Program at Animal Sciences Institute (NARC) is to study the emerging & re-emerging diseasesof livestock through use of molecular and epidemiological tools, improved diagnosis and devisingcontrol strategies based upon the local epidemiology of the pathogens . During the period underreport, most of the research activities aimed at studying the pathogenesis & pathogencharacterization, diagnosis and control of trans-boundary viral pathogens (Foot and Mouth Disease,Peste des Petits Ruminants) ; timely diagnosis of warble fly infestation to implement disease controlmeasures and surveillance of selected zoonotic diseases (Brucellosis and Crimean Congohemorrhagic fever) . This Program is also serving as hub for activities related to the Department ofAnimal Genomics and Biotechnology under the degree awarding institution of PIASA (PARC Institutefor Advanced Studies in Agriculture) at NARC .

a.

Control and Management Strategies Against MajorTrans-boundary Animal Diseases

Project 1 : Carrier potential of small ruminants in the persistence and transmission of pestedes petits ruminants (PPR) virus.

Following acute phase of the disease, Pestedes Petits Ruminants (PPR) virus shedding has beenreported for 11 weeks post recovery and in some animals even up to 12 weeks . It has also beenreported that animals infected by PPR virus are incubatory carriers and virus shedding wasdetected in their secretions and excretions 2 -3 days prior to the onset of the clinical disease .Under given conditions these carriers contribute to the unnoticed transmission of the disease atthe high risk areas like weekly livestock markets and communal grazing pastures . Suchasymptomatic carriers are threat to livestock farmers and may be responsible for trans-boundaryevents because the disease control measures cannot be instituted . A study was conducted toascertain the persistent shedding of PPR virus in the animals that experienced PPR and recoveredunder field conditions . Longitudinal data were collected by following five outbreaks where theanimals were sampled after 5, 10, 15 and 18 weeks post recovery. PPR virus shedding wasdetected 15 weeks post recovery. This evidence was supported by RTPCR, Efforts are underwayto recover the PPR virus from fecal material of such animals . Furthermore, all the samples will beexamined by rRTPCR to quantify the viral load .

Salient Achievements

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Hem-agglutination (HA) assay for the detection of PPR virus from persistently infectedgoats was optimized .

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One step PPR virus specific RTPCR, using F and N genes was standardized .

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Evidence of persistent PPRV shedding by recovered goats and sheep was recorded 15weeks PPR post outbreak . This evidence was supported by HA test and PPRV F-genespecific RTPCR .

Future Thrust

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Sampling will continue to be carried out in high risk areas (livestock markets, pastures forcommunal grazing) to strengthen the evidence of PPR persistence .

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Optimization of rRTPCR to quantify the viral load in persistently infected animals .

Project 2: Development of local isolate-based vaccine and strategic model for the control ofpeste des petits ruminants (PPR) in Pakistan

Pakistan is second to India in small ruminants' production in South Asia . The small ruminants areusually kept by low income landless villagers to whom this activity provides the main source ofincome . These rank second to poultry in the livestock species ladder while prioritizing the specieskept by the poor . The cost of a national outbreak of PPR was estimated to be worth Rs . 20.53 billionannually under an ALP funded project . The parameters quantified included losses due tomortalities (Rs . 11 .6 billion), meat loss (Rs . 7.13 billion), milk loss (Rs . 0.6 billion) and cost oftreatment (Rs . 1 .2 billion) . The magnitude of economic losses due to PPR in the country warrantmeasures to be taken to control this menace . This project was, therefore, initiated with theobjective to improve the socioeconomic conditions of small ruminants producers and achievingfood security through progressive control of PPR in Pakistan .


Two stake holders' workshop were successfully planned and organized at Azad Jammu andKashmir (AJK) Coordinated Unit in Bhimber district (sub-district Barnala) .

Eight suspected PPR outbreaks were investigated in Punjab and AJK . The outbreaks wereconfirmed by HA test, Ic-ELISA, cELISAand RT PCR .

Detection of PPR virus from the samples collected on filter paper by RTPCR was optimizedwithout RNA extraction .

Future Thrust

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Stake holders' workshop will be organized in all provincial and AJK Coordinating Units .

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PPR vaccination will be conducted in target tehsils .

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Outbreaks will be investigated throughout the country.

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Studies on the development of thermo-stable PPR vaccine will be conducted .

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PAKISTAN AGRICULTURAL RESEARCH COUNCIL


Future Thrust

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Development of ELISA for the detection of warble fly infested animals .

ANIMAL SCIENCES

Project3: Improving the diagnostic assay for the control of warble fly in Pakistan : Need andjustification

Warble fly is an economically important pest of cattle and goats in hilly, semi-hilly and sandy areasof Pakistan . The major economic loss inflicted to livestock and leather industry by the larvae of thisfly result from perforation in the hides and skins. Degradation in meat quality, poor weight gains ingrowing calves/kids and lowered milk production in lactating animals are the other economiclosses caused due to this disease. Economic losses of Rs . 20.6 and 2 .2 million due to warble flyinfestation in cattle & buffaloes in D.G . khan & Rajanpur districts, respectively have been reported .Diagnosis of warble fly infestation in live animals is based either on observation of clinical signsduring winter or serological methods. Direct clinical examination, generally results in anunderestimate of infestation level unless animals are examined regularly throughout theemergence period . Therefore, there is a dire need to establish a reliable serological diagnostic testfor diagnosis of warble fly . The project was initiated with the objective to determine the efficacy ofELISA for diagnosing warble fly infestation in local cattle and goats using local isolates ofHypodermalineatum and Przhevalskiana silenus .

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Field blood sampling for the whole year has been completed from selected experimentalanimals stationed at BLPRI, Kherimurat . In total, 1920 cattle and 1740 goats were sampled .

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Antigen has been prepared from first stage larvae of warble fly for the use in ELISA]

" Determination of accurate timing for the treatment and effective control of warble flyinfestation .

Project 4: Epidemiology, pathogen characterization and pathogenesis of foot and mouthdisease virus (FMDV)

Foot and Mouth Disease (FMD) infection puts a serious constraint on the economy andproductivity of the livestock in Pakistan . Understanding of FMD epidemiology andpathogenesis is crucial for FMD control efforts. The lack of epidemiological data like theprevalence, incidence, risk factors, transmission and persistence patterns of FMD in livestockand vaccine efficacy in the field is a serious impediment in the control effort . Therefore,following four studies are being undertaken to understand the epidemiology of FMD in Pakista

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Characterization of local isolates of FMDV and development of vector based vaccines .

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Epidemiology, vaccines and control of FMD in Pakistan .

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Epidemiology, pathogenesis andvaccine standardization forFMD in Pakistan .

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Setting up of Epi Node at AHP (Animal Health Program) under FAO project on FMD control .


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Established state of the art virology lab in Animal Health Program with the facilities of Indirect

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Established abio-secure animal house facility at Animal Health Program .

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Established Federal Epidemiology Node atASI, NARC .

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FMDV sequencing protocols were standardized and partial sequencing (VP1) of 42 selectedFMD field isolates collected during the years 2007 to 2010 and three vaccinal strains werecarried out. Variation is observed in FMDV serotypeOandA, while Asia 1 is stable .

Future Thrust

Sandwich ELISA, NSPELISA, Liquid Phase Blocking ELISA, conventional PCR, real time PCR,VN, LFBK cell culture and sequencing . All these tests were standardized at NARC andharmonized with PIADC .

The frequency of FMD outbreaks reported from various regions of Pakistan and FMD virusserotypes identified in these outbreaks during 2007 to 2012 . Error! Reference source notfound.FMD virus serotypes A, O and Asia 1 are identified serotypes with O being mostprevalent . Majority of these outbreaks have been reported in early to mid winter, owingpossibly to improve reporting andsurveillance network.

Vaccine matching studies were carried out using Merial and Arriah vaccines . The pooled seraobtained from Arriah vaccine reveled that serotype O has good matches, while A and Asia 1have not good matching with circulating strains in Pakistan . This will lead to the selection ofappropriate vaccine strain for FMDV control in Pakistan and developing regional FMDVvaccination programs .

Results of the persistence studies indicated that the Asian Buffaloes are carrier of FMDV Virusshedding was observed in oro-pharyngeal (OP) fluids and milk collected from buffaloesindicating the possible role of apparently healthy buffaloes in spread of FMD.

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Over 600 outbreaks were reported from district Karachi only, suggesting a highly endemicarea .

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The project has been successfully completed and the remaining planned activates will becovered in new project entitled "Epidemiology, vaccines and control of FMD in Pakistan"

" In future, anatomical pathology of persistence of FMDV in buffaloes and cattle will beconducted at a bio-secure facility, which is being constructed at animal health program,NARC .

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Federal Epi-node on FMDwill be made functional for data management andadvising in future .

b . Epidemiology and Diagnosis of Zoonotic Diseases One Health Approach

Project 1 : Establishment of Field Epidemiology Laboratory Training Program (FELTP) forjoint-human/animal disease investigation against Brucellosis, Crimean Congohemorrhagicfever and avian influenza

In the past few years, emergent disease episodes have increased globally . The list of importantemergent diseases is impressive indeed and, given what we know about disease ecology, it willonly continue to grow . Nearly all of these emergent disease episodes have involved zoonotic

infectious agents : the agents transmissible from animals and wild life to humans. The countriesthat have a national plan for the control of specific zoonotic diseases based on the availableresources and strategies are able to eradicate many such diseases . In spite of the progressachieved, the diseases like tuberculosis, Crimean Congo hemorrhagic fever (CCHF),Leishmaniasis, brucellosis and rabies still constitute a threat to human health and welfareparticularly in Asian region . The same is true for Pakistan . Therefore, there is adire need in Pakistanto understand the burden and epidemiology of zoonotic diseases in different regions of thecountry in livestock and poultry among high-risk groups of the population . Besides this, the riskfactor associated with these diseases needs to be studied so as to plan and implementappropriate interventions within the livestock sector, animal-human interface and the humanpopulation . The priority zoonotic diseases of animals selected for this study currently includeBrucellosis, CCHF and Avian influenza .


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Acountrywide network has been established for the surveillance of brucellosis and CCHF Inthis regard 19 provincial surveillance sites (PSS) have been set up.

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Atotal of 1399 serum samples of livestock origin were received from these PSS out of which342were found positive for brucellosis by Rose Bengal Plate Test .

" A zoonotic disease desk has been established to cater needs of data collection, collation,analysis and interpretation on zoonotic diseases under the Federal Epidemiology Unit atAnimal Health Program, NARC, Islamabad .

Future Thrust

Training

ANIMAL SCIENCES

B . millitensis isolated from afield outbreak in a sheep and goat flock (first isolate in Pakistan) .

To expand and strengthen existing surveillance network for brucellosis and CCHF

Capacity building of field staff and farmers on epidemiology, disease reporting and outbreakinvestigation and awareness of farmers especially women aboutzoonoses .

Joint vet-med meetings for collaborative work on human-animal interface to achieve OneHealth Goal .

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Training of two field staff were arranged in which 27 provincial field veterinarians were trainedin the area of surveillance, awareness and reporting and data collection of selected zoonoticdiseases .

Research Activities

ANIMAL NUTRITION

Inadequate feed resources and imbalanced conventional feeding practices are major causes of lowlivestock productivity in Pakistan . Major feed resources are crop residues, green fodder, andconcentrates . The availability of green fodder is limited in peri-urban commercial herds forcing thesefarmers to rely on crop residues and concentrates . The concentrates including cereal grains, industrialby products, oil seed cakes/meals, and molasses are very expensive and used mostly in smallquantities, resulting in inadequate nutrient provision to the animals . In this scenario, the Animal NutritionProgram at Animal Sciences Institute NARC focuses research primarily on the development ofappropriate and cost effective feeding systems/tech-nologies for farmers and commercialentrepreneurs . In this regard, the priority areas include efficient utilization of crop residues andagroindustrial by-products, and exploration of the latest developments in feeds and feeding . Having awell-equipped laboratory on nutritional analyses and highly trained scientific personnel, this Program isalso striving for human resource development regarding livestock feeding . The laboratory facility andexpertise are also extended to provincial research institutes, farmers and small-medium feed industry.

1 .

Effect of Stair-step Feeding Scheme on Weight Gain, Nutritional Input, Feed Efficiency andReproductive Performance of Buffalo and Sahiwal Heifers :

In stair-step nutritional regimens (SSNR), a period of growth restriction followed by a period of growthcompensation are imposed on growing heifers . Objective of SSNR is to reduce dietary energy input fora limited time and then compensate this effect by supplying excess dietary energy to exploit the effectof compensatory growth .

Feed cost represents about 60% of the total input costs on heifer rearing . Therefore, managing heiferswith less feed inputs by taking advantage of compensatory gains may have economic advantages .The SSNR could particularly be useful for buffaloes in Pakistan as their average age at puberty is 36months (1 '/1 year more than exotic and crossbred cattle) . In previous years, the ANP already hasshown that SSNR results in reduced feeding cost (by 11%) in raising heifers from 7 to 25 months ofage, and this is without any negative impact on growth rate, time to puberty, and conception rate . In thepresent experiments, effects of SSNR on development of Nili Ravi and Sahiwal heifers were studied .For this purpose, heifers of around 14 months age from both breeds were raised either on NBC'sdietary recommendations throughout the experimental duration (control), or on the stair-step regimen .The stair-step feeding procedure was to raise the heifers for 4 months on 80% of the NBC's energylevels followed by 2 months feeding on 120% of the NRC recommended dietary energy, followed bystarting the heifers on 80% NRC recommendation again and vice versa . Results regarding Sahiwalheifers are presented in table 1 .Table 1 . Performance of Sahiwal heifers fed control versus stair-step nutritional regimen

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°AKISTAN AGRICULTURAL RESEARCH COUNCIL

Parameters GroupsControl I Stair-step

-,'I re TI . ; ~- h

Avera . '1[1i"Z.zi . - W , " 196 .17 194.17Avera e total weight after 196 days (k) 344 .17 335.62Average daily weight gain (k) 0.796 0.686Daily D ry Matter intake (kg/head) 5.728 5.500Feed conversion ratio (kg of feed/kg wt gain) 7.196 8.017Heifers attaining puberty (number) upto 24 months 4(67%) 5(83%)M ature BW at first oestrus upto 24 months (%) 98 .33 95.89Savings over control (Rs .) - 2138

In case of Nili Ravi heifers, comparatively little higher weight gain (0 .565 vs . 0.536 kg/head) and betterfeed efficiency (10.35 vs . 10 .74) was observed in heifers of control group compared to stair-stepheifers however, dry matter intake in both groups were almost same (5.85 vs . 5 .76 kg/day) . No buffaloheifers showed attainment of puberty up to the age of 22 months .

2 . Evaluation of Bio-available Phosphorus in Indigenous Feed Stuffs in Poultry

About 60 to 70% of phosphorus (P) in plant feedstuffs is in the form of phytase P Non-ruminantanimals, such as poultry, lack significant amounts of endogenous phytase enzyme, which is requiredto separate P from the phytase molecule . Because phytase from plant feedstuffs is unavailable forabsorption, the unabsorbed phytase passes through the gastrointestinal tract, elevating the amountof P in the manure . Excess phosphorus excretion can lead to environmental problems . On the otherhand, ruminant readily use phytase because of the phytase produced by rumen microorganisms .

Need &Justification

Phosphorus availability incereal grains and vegetablemeals is low due to presenceof phytate . Availability of P inthese feedstuffs varies withvariety of plant and cultivationconditions of crops . InPakistan, available P values offeedstuffs have not beendetermined, so the values ofother countries are beingused by poultry feed industry .Exogenous phytase enzymeshave shown promising resultsfor the release of bound P inphytate complex . Bio-efficacyof the products needs to be

Figure 1 Sahiwal heifers in Animal Nutrition Program

ANIMAL SCIENCES

tested under local conditionsin Pakistan . Sixteen metabolic Figure 2. Chairman PARC (right) being briefed on the newly installed metabolic

cages for the project on bio-available phosphorus

trials on all the cereal grains, cereal by-products and vegetable meals will be conducted to determinebio-availability of P Six growth performance trials on broiler chicks will be conducted to determinecomparative bio-efficacy of commercial phytase enzymes.

SalientAchievements

This project is ongoing and in the first phase metabolic trials (Figure 1)on cockerels have beencompleted. The analyses regarding feed and fecal samples from the first phase is undergoing . In thisupcoming stages, growth trials on broilers will be conducted .

3. Preliminary Investigations on Risk Factors in Cottonseed Cake for DairyAnimals

The quality of cottonseed cake has been a cause of public concern during recent years. Many of thecottonseed cake batches produced in 2011/12 were reported to be refused by cattle/buffaloes . Insome instances, an alleged contamination of CSC resulted in total feed refusal and/or mortality atdairy farms . Present investigations were carried out as a preliminary attempt to validate the newsreports and understand the factors that might cause low quality of the cottonseed cake or it toxicity.

Need &Justification

Cotton is the main cash crop in Pakistan with an estimate of 2 .9 million tons harvest from 2.6 millionhectares for the year 2010-11 . The corresponding cottonseed cake production was approximately 2 .3million ton, making it the most abundantly available vegetable protein source for dairy animals . Anycontamination of cottonseed cake, the main ingredient in dairy feed, with dietary contaminants is notonly a health hazard to the dairy animals but also a risk for consumers via indirect contamination ofmilk .


Six samples of cottonseed cake associated with feeding problems, along with samples notassociated with such complications were collected . Stake holders were interviewed for gatheringhistory and symptoms of feeding problems associated with cottonseed cake . Results of the initialinvestigations are as under :

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The main complaint of the dairy farmers regarding low quality of cottonseed cake was feedrefusal by cattle/buffalo, along with reduction in milk production . At some farms, symptoms oftoxicity alleged to be due to cottonseed cake feeding included diarrhea or respiratoryproblems, which could be reversed afterwithdrawal of the cake feeding .

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Information collected from the stake holders indicated that low quality batches of thecottonseed cake are associated with several months of post-production storage, indicatingpossibility of fungal invasion during hot and humid climatic conditions .

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Cadmium exceeded the tolerable limits of 0.2 ppm in all the analyzed samples includingcontrols (0 .6-3.1 ppm in the suspected samples vs . 0.3-0 .6 ppm in the controls), while leadand chromium at toxic levels were also detected in samplesfrom one city.

Detailed analyses for mycotoxin contamination revealed high levels of aflatoxins andochratoxins in all the samples including controls . Overall, heavy contamination of the

AKISJAN AGRIC;L .IL-CUFiAL HESEARCN COUNCIL

Training/Extension Activities

March, 2013)

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Buffalo Research Institute (BRI) Pattoki (27th March 2013) .

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5600 bags of PARC Cattle Feed . Figure-2

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1500 Multi-Nutrient Urea-Molasses Blocks .

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350 sachets of Milk Booster (mineral premix) .

ANIMAL SCIENCES

cottonseed cake with toxigenic Aspergillus, Penicillium, Claviceps, Fusarium,Alternaria/Phoma, and Arthrinium spp was found . The novel finding in this regard was thepresence of tremorgenic mycotoxins in excessive quantities from Claviceps and Aspergillusspp. in the samples suspected of feed refusal and toxicity problems .

1 . Four seminars on "Effect of stair-step nutritional regimen on growth rate, nutrient utilization,pubertal age and conception rate in Nili-Ravi buffalo and Sahiwal cattle heifers" were delivered atfollowing venues :

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Institute of Animal Nutrition & Feed Technology, University of Agriculture, Faisalabad (14th

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Research Centre for Conservation of Sahiwal Cattles (RCCSC), Jhang (25th March 2013) .

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Livestock Production & Research Institute (LPRI), Bahadur-Nagar (26th March . 2013) .

2 .

A seminar on "Preliminary investigations on cottonseed cake implicated with feed refusal andcattle poisoning problems in Pakistan" was delivered in the 67th Conference of the Society ofNutrition Physiology, 19-21 March 2013, University of Gbttingen, Germany .

3 .

40 DVM students from various universities (PMAS-UAA, Rawalpindi, SAU, Tandojam, LUAWMS,Uthal) completed part of their internship at the Animal Nutrition Program .

4 .

The Feed Technology Unit of the Animal Nutrition Program has been involved in popularizingnutritionally balanced blended cattle feed and helping farmers and small-medium scale feedmillers in operating feed mills . Additionally sale of Multi-Nutrient Urea-Molasses Blocks to farmersin the areas with fodder scarcity has been a major activity. In this regard following are theachievements for the last year :

Figure 3. Different products produced at the Feed Technology Unit of the Animal Nutrition Program

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30 farmers and small-medium scale feed millers from various parts of the country were giventechnical assistance .

Future Thrust

Results regarding the stair-step heifer development for early and economical heiferdevelopment will be disseminated to the farmers and other stake holders .

Laboratory analyses and further metabolic trials will be conducted regarding the project onbioavailable phosphorus in indigenous feedstuffs in poultry.

Investigations will be continued on cottonseed cake samples associated with feedingproblems .

A research proposal on assessment of amino acid availability in locally available feedstuffs forchickens is under process .

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PAKISTAN AGRICULTURAL_ RESEARCH COUNCIL

ANIMAL REPRODUCTION

ANIMAL SCIENCES

Improvement in reproductive efficiency of buffalo and cow is the major objective of present researchactivities at the Animal Reproduction Program . Buffalo and cow are both important dual purpose (milkand meat) animals of Pakistan . Artificial insemination (AI) is the most important technique to improvegenetic potential of livestock herd in shortest possible time . First research area is to improve post-thawsemen fertility which is especially required in buffalo as the conception rate in buffalo with frozenthawed semen has been reported low in large scale field trials . It has been observed that improvedsemen preservation techniques resulted in rise of conception rate from 30 to 50 percent in buffalo .Continued efforts are being made to further improve these results .

Second research area is expanded application of AI with estrus synchronization coupled withimproved management practices . Due to scattered livestock population and small holdings (1 to 7animals per farmer), application of AI is not easy on large scale . Bringing large number of animals inheat at one time in a particular area will make application of AI in these animals convenient . Efficacy ofthis technique can be further improved by application of better management practices such as use ofmale effect called biostimulation . Furthermore, this line of action will be used to overcome seasonalityof breeding especially in buffalo .

Research Activities

a.

Improve Post Thaw Quality/Fertility of Buffalo Semen

Experiment 1 : Effect of bovine serum albumin in extender on post-thaw quality of buffalobull spermatozoa

During cryopreservation of semen up to 50% viability of sperm is lost and the main reason is theproduction of toxic reactive oxygen species (ROS) . Sperm cells are exposed to oxygen and visiblelight radiation during processing of freezing leading to the formation of ROS . Albumin is animportant extra cellular antioxidant minimizing the formation of ROS (OH- radical) . Objective of thepresent study was to determine the effect of adding bovine serum albumin (BSA) at two differentconcentrations (0.5% and 1 .0%) in tris citric acid extender on post-thaw quality of buffalo bullspermatozoa .


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Semen frozen in extender containing 0.5% BSAwas significantly better that that %vith 1 .00, BSAor control in terms of post thaw acrosome integrity

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Addition of 0.5% BSA in tris citric acid extender gave higher conception rate than control afterArtificial Insemination in buffaloes .

Experiment 2: Effect of duck egg yolk plasma in extender on post thaw quality of buffalo bullspermatozoa

Egg yolk from chicken and ducks has been used as a component of semen freezing extenders formost of the animal species including buffalo probably because of its easy availability . It ishypothesized that plasma from duck egg yolk can replace the whole egg yolk in tris citric acidextender as it has the potential to preserve the post thaw quality of buffalo bull spermatozoa . Thisstudy was designed to evaluate the effect of plasma from duck egg yolk (10°0 15% and 20%)

compared to control (whole duck egg yolk 20% or whole chicken egg yolk 20%) in tris citric acidbased extender on the basis of post thaw viability of buffalo spermatozoa.

Salient Achievements:

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The Computer Assisted Semen Analyzer (CASA) motility parameters were better in extendercontaining 15% plasma from duck eggyolk although the difference was not significant

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Whole duck egg yolk or whole chicken egg yolk can be replaced with 15% plasma from duckegg yolk in tris citric acid based extender for cryopreservation of buffalo bull spermatozoa.

B. Improving Reproductive Efficiency of Cows and Buffaloes Through Estrus Control andBetter Management

Experiment 1 : Effect of biostimulation during estrus synchronization on conception rate inbuffalo

Reproductive efficiency is one of the primary factors affecting productivity of buffaloes . Hormoneshave been used over last two decades to stimulate reproductive system in cattle . However use ofhormones without improved management practices like nutrition, housing and bio-stimulation willnever be able to stimulate animal body to achieve the desired objectives . Bio-stimulation in theform of presence of a bull in the herd is considered beneficial to keep ovaries active and bringanimal into heat . Furthermore there are indications that biostimulation results in improvedconception rate in cattle . Present work was conducted to assess the effect of biostimulation onfertility in buffaloes treated for oestrus synchronization with GnRH + PGF2a (ovsynch program)with an assumption that conception rate will further improve

Salient achievements

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A better conception rate was achieved after oestrus synchronization in the presence of male(biostimuilation) as compared to thatwith (Figure-3) no biostimulation (Figure-4) .


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Six training courses (two week each) were conducted for communityworkers from Punjab andBalochistan on artificial Insemination and pregnancy diagnosis in cattle and buffalo (totalparticipants = 90) .

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Imparted training to community workers from Sindh, Gilgit-Baltistan and Chitral on Moderntechniques of livestock production

Forty DVM internees from Veterinary faculties of Balochistan, Sindh . AJK, Punjab and KPKAgricultural Universities were given practical training in the latest Animal reproductiontechniques

Future Thrust

FourM.Phil students conductedthesis research

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Investigations will be carried out on Fertility-Related Biomarkers in Buffalo Semen to ReduceMale-Factor Losses

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-'AKISTAN AGRICULTURAL RESEARCH CCIUNCII_

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Effect of presence of bull will be studied during summer season in buffalo

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Work will be conducted on fertility of Sahiwal cows after estrus synchronization at farmer level

Figure 4. Buffaloes synchronized for estrus withovsynch (GnRH + PGF2a)

ANIMAL SCIENCES

dmc0

.aduCOV

Oestrussynchronizationalongwithbiostimulation

Oestrussynchronizationwith nobiostimulation

Figure 5. Conception rate in buffaloes treated withovsynch program for estrus synchronization in thepresence or absence of bull

DAIRY TECHNOLOGY

Figure 6 . Dairy technology at NARC

Each year about 47 billion liters of milk is produced in Pakistan . Around 15% is spoiled or remainsunder value at source/farms . Hardly, 10% is collected properly and milk plants process about '/a of this(1 .2 billion liters) . The rest (90%) is processed by small-scale traditional technologies (100 to 500 literbatches) which are generally inefficient and being primitive lack quality and hygiene thus posing healthhazards and draining farm economy. These processors do not have access to trainings and often finddifficulties in getting the right kind of information to prepare and sell quality dairy products .

The Dairy Technology Program focuses on problems of smallholders / producers and /or processors,trying to find better alternates of indigenous technologies and make them suitable and provides result-oriented trainings on hygienic milk production and cottage scale dairy product preparation . Theultimate target is value addition to milk by expanding dairy product variety and thus business therebygenerating better employment opportunities for small investors both in rural and urban areas .

Research Activities

a.

Farm Milk Quality

Experiment 1 . Evaluation of farm milk and preparation of valueadded dairy products

Although Pakistan stands first in milk production among Muslim World and 4th in overall worldranking and is presently producing about twenty 47 million tones of milk annually, yet quality of themilk and milk products is not satisfactory which is generally due to poor management techniquesbeing practiced by dairy farmers . Apart from milk quality, the quantitative production is also lowandin due course oftime average animal productivity with respect to milk quantity is declining . Feed andgenetics are important limiting factors . How these factors are affecting different milk components,such as milk fat, protein etc and other milk physiochemical characteristics is not known with respectto our milch breeds . These are not only important as economic traits of animals but at the same timequality of milk and milk products and hence the over all farm income and economy.

Herd testing is a tool of better management of dairy farm and the best way of testing is milk testing.This involves collection of individual milk samples from each milking animal in a herd . The volumeof each milking-produce is recorded and tested for milk fat, protein, lactose, minerals, density, etc.This year focus of Program was to help NARC Dairy Farm Management, by evaluating farm milkquality as it is a window to the production of individual animal in a herd to run their farm moreefficiently . Furthered to that, value added dairy products, particularly spicy cheese & yoghurtvarieties and relevant training to small-holders / internees for diffusion of knowledgewere also partof this undertaking .


ANIMAL SCIENCES

"

Comparing on average content basis, buffalo milk was found richest in fat (6.0%) and milksolids (14 .6%) while cow milk was lowest at 4.4 and 13.1 %, respectively (Tables 2 & 3) . Theaverages stated met the defined respective standards but did not match real/normal fieldvalues particularly SNF content of buffalo milk .

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Thecorresponding protein values of both types of milk are somewhat within normal limits butash contents are significantly lower in buffalo milk which also reflected from low SNF contentsof buffalo milk . Buffalo milk is also low in lactose contents . On overall quality basis buffalo milkwasfound relatively poor than that of normal field values .

Table 2 .Percent milk composition (Means ± SD) of selected cows of NARC Dairy Farm

Tag No . N Total Solids Fat SNF Protein Lactose Ash143 C 2 11 .6--0 .1 3 .3-0 .1 8.3±0 .1 3 .0±0 .1 4 .6±0.1 0.7±0 .1150 C 4 12 .1±0.3 3 .9±0 .2 8.2±0 .3 3 .0±0.1 4 .5±0.2 0.7±0 .1168 C 3 13.0±0 .4 4.2±0 .4 8.8±0 .1 3 .2±0.1 4 .8±0.1 0.7±0 .1158 C 5 13 .2±0.5 4.4±0 .6 8.8±0.2 3 .2±0.1 4 .9±0.1 0.7±0 .1387 C 4 13 .2±0.9 4.6±0 .9 8.6±0.3 3 .2±0.1 4 .7±0 .2 0 .7±0.1147 C 5 15 .4±0.8 5.9±0 .8 9 .5±0.2 3 .5±0.1 5 .2±0 .1 0 .8±0.2Grand Mean ± SD 13.1±0.5 4.4±0.5 8.7±0.1 3.2±0.1 4.8±0.1 I 0.7±0 .1

"

Animal ranking/comparison, according to milk quality, has been shown in Figures 5 and 6which revealed quiet clearly low quality milk producing animals which should be culled /replaced and which one be retained and taken care better for enhanced productivity. Thedetailed milk composition comparison helped to predict animal productivity and thus serve aspowerful tool in farm management .

"

A variety of dairy products was produced and evaluated by a taste panel . It was observed thatbuffalo milk was of best-quality both as fluid milk and in dairy products made there of i.e .,cheese and yoghurt .

Trainings/Extension/Technical Services

Training, extension and technical services are hallmark of Dairy Technology Program which areextended to farmers (individually or in groups), cottage business persons, small producers orprocessors and university graduates . In the year under report major focus of training was studentinternships which included both class room instructions and hands-on practice on milk analysis andproduct preparations ; a total 1225 man-hours on training was spent .

"

Trainings and extension services were extended to farmers (individually or in groups), tostudents (as internee or class room teaching) . women farmers / activists or house wives . Someof the trainings were organized in collaboration with some agencies such as AgriculturePolytechnic Institute (API)

"

Training & tutorship was also provided to students of UAAR / WAS (PIASA) .

"

Many progressive farmers were given advice / training on clean milk production and on on-farm milktesting etc .

Future Thrust

Participated in four Agri Exhibitions and displayed various dairy products for public awarenesson milk and milk product quality and to convey information about training opportunities in thisfield at NARC .

Studies regarding development of suitable non-gelatin based stabilizer for improvingconsistency and texture of yoghurtwill be conducted

Table 3.Percent milk composition (Means ± SD) of selected cows of NARC Dairy Farm

120

~-T 1 i

Tag No. N Total Solids Fat SNF Protein Lactose Ash1,52 -B 2 13 .8±1 .1 5 .,5-1 .4 8 .3±0 .2 3 .9±0 .1 3.7-0 .1 -0.6-0 .1454 -B 5 14 .0±1 .0 5 .4±0.7 8 .6±3 .3 4 .1±0 .2 3.8±0 .2 0.6±0 .1547 -B 4 14 .3±1 .3 6 .0±1 .1 8 .3±0 .2 3 .9±0 .1 3.7±0 .1 0.6±0 .1183 -B 3 14 .7±1 .8 6 .0±2.2 8 .7±0 .4 4 .1±0 .2 3.9±0 .2 0.6±0 .21461 -B 2 14 .9±0.3 6 .0±0.2 8 .9±0 .1 4 .2±0 .1 4.0±0 .1 0.6±0 .13 5 -B 5 14 .9±0.5 6 .1±0.2 8 .8±0 .4 4 .0±0 .1 4.1±0 .1 0.6±0 .1495 -B 2 15 .0±1 .3 6 .4±1 .1 8 .6±0 .2 4 .1±0 .1 3.8±0 .1 0.6±0 .2539 -B 3 15 .3±1 .1 6 .6±1 .5 8 .7±0 .4 4 .1±0 .2 3.9±0 .2 0.6±0 .1Grand Mean ± SD 14 .6±1 .1 6.0±1.0 8.6±0.7 4.1±0.1 3.8±0.1 0.6±0.1

16 .0

14 .0

12 .0

10 .0

6.0

6,0

4.0

2,0

0.0539-B 152-B 183-8 188-B 35-8 461-B 454-B 495-B 547-B

Tag No . of buffaloes

Total Solids

0 Fat

ESNF 0 Protein

ELactose

Ash

0

Continue offering trainings

ANIMAL SCIENCES

149-c 168-c 150-c 147-c 387-c 158-cTag No . of cows

Total Solids " Fat

0SNF

Protein ELactose

Ash

Figure 7: Percent composition of fresh milk of selected

Figure 8: Percent composition of fresh milk of selectedbuffaloes at NARC Dairy Farm

cows at NARC Dairy Farm

.

Research work on "identification and authentication of market meat and meatproducts using molecular approaches" has been planned .

"

Continue participation and dissemination of research-based information andtechnology through exhibitions, publication and trainings etc.

Pakistan is rich in livestock resources especially ruminants . Currently there are about 28.4 millionsheep with 28 well know breeds and 63 .4 million goats with 25 well known breeds in Pakistan . SmallRuminants are producing about 0.629 million tons of mutton and 0.775 million tons of milk annually. Inaddition to supply of proteins of high biological value, these animals are also contributing significantlyin the socio-economic aspects of national economy. Small Ruminants play an important role in thelivelihood of landless and small farmers as well as in the overall national economy. Small RuminantResearch Program (SRRP), Animal Sciences Institute, NARC strives for the improvement of animalfood products (e .g ., mutton and milk) through research endeavors on small ruminant production . TheProgram has the facilities of rearing and experimentation of small ruminants . In countries with advancewool producing and processing industries objective measurement of the physical characteristics ofwool is an important tool to determine price and predict processing performance of a particular lot ofwool, similar opportunities are available at the Wool Testing Service of the Small Ruminant ResearchProgram, ASI, NARC . Whether, one needs objective information to produce good quality yarn, checkthe quality of imported or locally produced wool or wants to carry out research on wool quality, theWool Testing Service, NARC may be helpful .

Research Activities

SMALL RUMINANTS

a. Exploiting Fattening Potential to Improve Mutton Production

Project 1 . Study on production potential of different sheep and goats for mutton productionunder high input system"

Mutton production gap between the conventional farming system and commercial farmingsystem is significantly wide . Under traditional farming system of Pakistan, small ruminant's weightgain is often low, however daily weight gain under balanced feeding and management programcan be improved significantly . Improved system of production like energy and proteinsupplementation increases the production performance, survivability and growth rate in smallruminants . Therefore need was felt to investigate the potential of sheep and goat breeds forcommercial meat production on high input system rather than extensive system . This study wastherefore designed to see new avenues of mutton production from small ruminants . In this regard,fattening trials on three groups of male Thalli sheep, by allocating three feeding regimen, wereconducted and results are shown in table 4 .

Table 4 . Carcass evaluation of male Thalli sheep

122

-'AKISTAN AGRICULTURAL RESEARCH

Parameters Group 1

(Concept rate+ grazing)

Group 2 Group 3(100 % VCR (Concentrate ad

libitune ++ grazing) grazing)

No . of animals slaughtered 3 3 3

Average live weight before slaughter (kg) 68.8±8.31 49 .7+2.57 56.0±8 .26Average live weight after slaughter (kg) 66.3±8.31 48 .0+1 .80 53 .8±8.01Carcass weight (kg) 34.4±2.98 24.5±1 .24 27 .2±2.93Dressing (%) 50.0 49 .2 48 .5Blood (kg) 2.5±0.00 1 .7±0.76 2 .2-0.29

Parameters


"

On the basis of above referred successful trials, the Program in collaboration with RangeResearch Program . fattened fifty male goats purchased by PATCO for sale at the occasion ofEid-ul-Azha .

"

ThePATCO earned hundred thousand rupees as profit from the sale of these animals


Meetings

"

The PI . RADP-ASD Sub-Project "Study on the production potential of different sheep and goatbreeds for mutton production under high input system" attended the Mid-Year Review Meetingof RADP Sub-Projects (2012-13) held at Committee Room, NARC on March 27, 2013 .

"

Dr. M.N .M . Ibrahi m and Dr. Nile from ILRI had a meeting with Member (ASD), PARC on 24-04-2013 . They also held meetings with AIP (Livestock Component) focal persons (ProgramLeader. SRRP being the focal person on small ruminants) .

ANIMAL SCIENCES

The Program Leader, SRRP attended the meeting of Agricultural Innovation Project (AIP) heldin Islamabad on March 9, 2013.

The Program Leader, SRRP has been appointed as Country Coordinator (Pakistan), AsianAustralasian Dairy Goat Network (AADGN) of Malaysia .

Seminars

"

The scientists of SRRP attended a special seminar entitled 'A Conversation on Challenges

12 3

Skin (kg) 6.3=0.15 4 .8=0 .35 3.0-1 .00

Stomach with ingesta (kg) 8.2±0.76 5 .3±1 .10 6 .2~ 1 .78

Stomach empty (kg) 1 .3±0.37 1 .3±0.26 1 .2±0.26

Intestine with ingesta (kg) 5.1±1.71 3 .2±0.25 3 .2±1 .08

Intestine empty (kg) 3.4=1 .01 2.0±0.68 2 .4±1 .08

Head (kg) 4.50.35 3 .2=0.19 3 .5±0.30

Liver (kg) 1 .1±0 .17 0.8-0.00 0.910.10

Heart (kg) 0 .206 0.186 0.233

Left Kidney with fat (kg) 0.367 0.260 0.447

Right Kidney with fat (kg) 0.247 0.210 0.333

Left Kidney without fat (kg) 0.037 0.040 0.093

Right Kidney without fat (kg) 0.040 0.043 0.105

Lungs (kg) 0.8=0.20 0.6-0.58 0.7±0.12

Fat (kg) 4.2±0.21 5.3-0.42 3.9 -1-1 .76

Group I Group 2 Group 3

(Concent rate (100 % NCR (Concentrate ad

+ grazing) + grazing) libitum + grazing)

Trainings

"

Program Leader (SRRP ASI) attended the one-day workshop on Peste Des Petits Ruminants(PPR) in Sheep and Goats on September 18, 2012 at NARC.

"

Program Leader (SRRP) gave lectures to the course workshop of M .Sc . (Hons .) LivestockManagement atAllama Igbal Open University, Islamabad on Sep . 27, 2012 .

"

A delegate from Small Industries Wing, Government of Balochistan visited Wool Lab, NARCon November 19, 2012 for training on wool analysis .

" The SRRP imparted two-weeks practical training on small ruminant management andpractices to the Internees-DVM students of Sindh Agriculture University, Tandojam duringFebruary, 2013 .

"

Program Leader (SRRP) attended the Workshop arranged by Ministry of National FoodSecurity & Research at NARC on February 20, 2013 .

"

The Program Leader, SRRP delivered lectures on livestock management to the M .Sc . (Hons .)students of Allama Igbal Open University on March 18, 2013 .

"

The Senior Scientific Officer, SRRP gave a lecture to a class of Livestock Management onBiometry at Conference room of Animal Nutrition Program, ASI on March 20, 2013 .

Visits

Associated with Global Food Security Focusing Animal and Agriculture" on April 11, 2013 atthe Conference Room of IABGR (PGRI), NARC delivered by Prof . Dr. Muquarrab A. Oureshi(NIFA/USDA) .

The DVM (Final Semester) student internees from University of Rawlakot, Azad Kashmir weretrained on small ruminant production fortwoweeks .

The SRRP imparted practical trainings on small ruminant management and practices to thedifferent batches of Internees i .e ., DVM students of SAU, Tandojam, Pir Mehr Ali Shah-University of Arid Agriculture, Rawalpindi and UVAS, Lahore during March, 2013

The Program Leader (SRRP ASI) attended the consultative workshop on understandingdrivers of ecosystem change and livelihoods in the upper Indus basin on April 29-30, 2013held in Islamabad .

"

The Program Leader, SRRP attended the training workshop on "Financial ManagementSystem of PARC" on May 6-10,2013 at NARC.

"

TheSSO SRRP gave lectures & practical trainings to the class of Livestock Management onAnimal Breeding at Livestock Research Station, ASI in April and May, 2013 .

"

A group of L&DD officers, Government of Punjab visited NARC on July 03, 2012 . Sheep &goats were displayed at Livestock Research Station for their study-visit .

"

Dy. Director (Planning), Government of AJ&K visited Small Ruminant Research Program forguidance on developing a PC-1 on exotic dairy goat breed .

124

-~AKISTAN AGRICULTURAL RESEA

"

DG (Research) KPKvisited SRRP on September 18, 2012.

Future Thrust

ANIMAL SCIENCES

Mr. David W. Doolan, International Project Manager, FAO metwith Program Leader (SRRP) .

Program Leader (SRRP ASI) visited Jaba Sheep Farm and participated in the Small RuminantsManagement and Disease Control Workshop in November, 2012.

"

The Dean, Faculty of Animal Husbandry and Veterinary Sciences, University of Agriculture,Peshawar, KPK visited SRRP on December 13, 2012 .

"

Mr. Shafi Ullah, Director, Mountain Agricultural Research Center, Gilgit visited Small RuminantResearch Program on January 31, 2013,

Dr. Dilpazeer, Livestock Department, Azad Jammu & Kashmir visited Small RuminantResearch Program on February 06, 2013 .

"

Dr. Chaman Lal, Lecturer, Allama Igbal Open University visited SRRP along-with students towitness the small ruminant research facilities at NARC on March 19, 2013 .

Forty-seven farmers visited/contacted SRRP for guidance on small ruminant managementand production during the current year.

Publications : Journals, Proceedings, Books, Extension Articles

"

Khan, M .F and F Ashfaq, 2012. The Recent status of dairy goats in Pakistan with specialreference to their potential, opportunities and challenges faced by scientists and farmers . Pak .J . Livestock Sci . Vol . IV.

"

Khan, Saeed-UI-Hassan, M .N . Riaz, A . Ghaffar and M .F. Khan, 2012 . Calpastain (CAST) GenePolymorphism and its Association with Average Daily Weight Gain in Balkhi & Kajli Sheep andBeetal Goat Breeds . Pakistan J . Zool . Vol .44 (2), pp : 377-382 .

" Khan, M .F. and F Ashfaq, 2012 . Dairy Goats in Pakistan : Potential, Opportunities andChallenges . In Proceedings : 1 st Asia Dairy Goat Conference, University Putra Malaysia, KualaLumpur, Malaysia, 9-12 April, 2012 . pp : 60-64 .

"

Improvement of local goat breeds through crossbreeding with Boer goat .

"

Studies on improvement of sheep and goat production .

Research Activities

Salient Achievement

POULTRY DISEASE CONTROLPoultry sector in Pakistan is one of the fastest growing sectors of livestock segment of agriculturebased economy of the country. A steady annual growth of 10-12% has been observed since 1990 inthis sector. However, one of the limiting factors in further growth in this sector has been the intermittentintroduction of infectious diseases . Avian health is major concern for the farmers and backyard poultryowners in the wake of high prices of poultry meat produced in Pakistan . Especially with the previousintroduction of bird flu H5N1 in this country during 2006-08, and its subsequent threat due to itscontinuous circulation in other countries of the region, need for food safety and food security hasincreased manifolds . Since 2003, Avian Influenza viruses of different types have been severelyaffecting the growth of commercial and backyard poultry . With the recent outbreaks of highlysignificant poultry diseases of zoonotic potential, enhanced diagnostic capacity at national level isneeded . Although various diagnostic kits, reagents, drugs and various vaccines are being importedfor the control of different poultry diseases, however, these drugs and vaccines are less effectiveagainst local strains of these poultry pathogens . Keeping in view the above scenario, the NationalReference Lab for Poultry Diseases (NRLPD) at the Animal Sciences Institute of NARC has beenextending surveillance, diagnosis and research activities in this country. The NRLPD is equipped withthe state of the art equipments, highly qualified and trained manpower and strong national andinternational collaborations in the field of avian disease diagnosis and research .

a .

Surveillance, Diagnosis and Disease Management Services Against Major Avian Diseases

To support nation-wide surveillance and referral diagnostic activities in Pakistan in the area of AvianHealth, a network of 10 labs (Figure-7) has been set up in different parts of the country in collaborationwith the provincial poultry production offices . Furthermore, due to the emergence of new strains ofhighly virulent avian pathogens in the country, efforts are required to undertake relevant research in thisregard . Above all there is a strong need to undertake capacity building of the federal and provincialinstitutions involved in surveillance, research and diagnosis of avian diseases . The current activities atNRLPD are destined to achieve all above .

"

The NRLPD provided nation-wide surveillance for avian diseases and obtained data forNational AI Surveillance . 11603 samples were processed and 73 isolates of various poultrypathogens (AIVs, H9N2, NDV, IBV, IBD, AAVs, ALV, MDV APV MG, MS, Pasteurella, E . Coli andSalmonella) were made .

"

Production of Diagnostic reagents and their provision to public sector lab was done . Thefollowing viruses recovered from the field were characterized and sequenced : 30 of NDV, 7 ofAIV-H9 and one H4N6 and sequences were submitted to international sequence data bank(Genbank)

"

5272 blood samples were tested for determining sero-conversion against variety of vaccinesused in commercial poultry.

"

In an experiment, differential pathogenesis and its transmission for AAV-4 was studied . Here25 isolates were studies and RFLP- PCR was standardized for AAV The data revealed that in

ANIMAL SCIENCES

AMWmost of the outbreaks of Hydropericardium syndrome 3-4 sub-genotypes of AAV-4 wereinvolved and co-infection of AAV-8 serotype was of major concern .

Vaccine trails against NDV and AN H9 were performed for evaluation of specificity of currentlyused vaccines of poultry pathogens, revealing significant antigenic variation of the circulatingfield viruses

"

Determined the Pathogenicity and drug susceptibility of a new bacterium, (Past . Haemolytica)from poultry, first time in this country.

"

Differential diagnosis was provided for ALV, MG, Salmonella, and Marek's Disease to majorGP companies in this country

"

Two national trainings having 24 participants were conducted for extending surveillanceprocedures for different Poultry diseases .

Two M . Phil students completed their research work and 53 internees were extended 4-6 weektraining each

"

Two posters were presented in International Microbiology conference in Karachi

- CENTRAL LAB/ NRLPDISLAMABAD

- PROVINCIAL LABS

O SINDHO PUNJAB 81SD

BALOCHISTAN

0 NWFP & FATANORTHERN AREAS

_ AJK

[MJ

Figure 9 :

Future Thrust :

"

Expand surveillance for poultry diseases in the country.

"

Conduct trainings for awareness, sampling and initial screening of major poultry disease .

"

Sequencing of new isolates of NDV, AIVs, IBV and AAV-4 .

"

Studypathogenesis of newsalmonellae.

"

Pathogenesis study on NDV isolates 2009-14.

b.

Epidemiology and Diagnosisof Zoonotic Diseases ONE HEALTH APPROACH

Project-1 : Establishment of Field Epidemiology Laboratory Training Program (FELTP) forjoint human/animal disease investigation against avian influenza

The emergence and re-emergence of zoonosis and their potentially disastrous impact on humanhealth is a growing concern around the globe . The ease and speed of international travel ; the rapidgrowth of international trade in poultry and poultry products due to globalization, urbanization ofpopulations, and dramatic changes in poultry production systems and patterns are important factorsin the spread of poultry diseases and zoonosis . Surveillance and the early diagnosis and detection ofpoultry pathogens are crucial components of disease control, prevention and eradication strategies,In Pakistan there is a strong need to understand the burden and epidemiology of Influenza disease indifferent regions of the country in poultry. live bird markets, wild birds and backyard poultry.

Afederal network encompassing all the provincial setup to coordinate such surveillance is expected tobe developed in future . In order to supportanyfuture surveillance activities, there is a defined need todevelop a trained veterinary workforce, reinforce laboratory capacity to test zoonotic influenzainfection samples, and influence the way the animal-human health surveillance system develops . Asuccessful system requires capacity at all three levels but particularly at the local and provincial levelwherediseases are first identified and reported, andwhere response must take place.


Atotal of 19 provincial surveillance sites have been set up consisting of Punjab (n=3), KhyberPakhtunkhwa (n=4), Baluchistan (n=2), Sindh (n=4), Azad Jammu and Kashmir (n=3),Gilgit-Baltistan (n=2) and Islamabad (ICT) .

"

In total 6842 samples of poultry origin were tested for AIVand 18

H9N2AIVs were isolated .

"

14 H9N2 AIVs were sequenced for HA and NA genesand cartography experiment was carriedout to evaluate immunogenic potential, revealing that more than 3 isolates showed distinctvariation in their antigenicity, requiring their including in newvaccines .

6 meetings have been arranged underone health concept project.

Two international research papers were published and two posters were presented inInternational ASM meetings .

c.

Regional Cooperation on Highly Pathogenic Emerging Diseases (HPEDs) in SAARCCountries under FAO project

To strengthen and empower SAARC countries in their ability to prevent, control and eradicate HPED,including Highly Pathogenic Avian Influenza (HPAI), through improved veterinary and public healthservices and inter-sectoral collaboration on a regional basis . To fulfill the mandate of the project,SAARC Regional Leading Diagnostic Laboratories (RLDLs) have been established in Bangladesh(PPR), India (FMD) and Pakistan (HPAI) as agreed by the member countries . These laboratories arecoordinating and leading a network of national diagnostic laboratories, primarily focusing to maintain

128

PAKISTAN AGRICULTUR/

uniform diagnostic standards, support training of laboratory scientists/technicians from the memberstates, and backstop regional surveillance and epidemiological studies . The activities are also beingsupported by the international OIE and FAO reference laboratories like Australian Animal HealthLaboratory, Geelong, Australia . In this regard the National Reference Lab for Poultry Diseases at NARChas been identified as RLDL for HPAI in this region .


"

After completing the required capacity building, the NRLPD is undergoing Accreditation forISO-17025 certification through UNIDO. In this regard application has been submitted andpre- assessment has been completed by PNAC-Islamabad

"

Referral diagnosis was extended to 682 commercial farmers and eight public institutions .

"

22H9N2 AIVs have been isolated .

"

OIE diagnostic protocols have been standardized at NRLPD and the technical manual hasbeen prepared accordingly .

"

Flock health monitoring and testing of post-vaccination immune status have been performedfor 948 farms

"

Seroprevalence against AIVs subtypes H5N1, H7N3, H4N6, H3N1 and H9N2 has beenperformed using 2660 serum samples received from non vaccinated birds .

"

Organized and conducted an international training for personals of National AI labs in theSAARC countries (Afghanistan, Bangladesh . Bhutan, India, Maldives, Nepal, Pakistan & SriLanka)

"

Organized two national trainings for harmonization of protocols where 12 persons from all overthe country participated

Future Thrust

"

Continue referral diagnosis services .

ANIMAL SCIENCES

Participation in international proficiency testing and provision of proficiency testing to SAARCcountries .

"

Extend trainings to staff from SAARC member states and those from other provinces ofPakistan .

Acquire international ISO-17025 certification for NRLPD.

Standardization of protocols including PCR, Real Time PCR and sequencing for diagnosis ofnew AIV subtype H7N9 .

"

Organize joint vet-med meetings for working in a liaison for joint activity on human-animalinterface .

Training of field staff in human/animal health and farmers in epidemiology, disease reporting,and outbreak handling/ investigation .

"

Standardization of reverse genetics technique for development of better vaccines

Poultry

Poultry production is considered to be one of the most efficient and economical system of productionof animal protein . In Pakistan Poultry has been playing a vital role in bridging the gap between supplyand demand of animal protein which is an essential food nutrient . Our poultry production is mostlydependent on exotic commercial lines but ever since the existence of this industry in Pakistan noattention has been given to our backyard poultry. Indigenous chickens, called desi, have theadvantage of being well adapted to the local environmental conditions (hot cold humid dry and rainyweather) . These chickens are scavengers by nature and a system of supplementary feeding is notpracticed . They scavenge on field grains, insects, earthworms, green matters, crop residues,homestead pickings and kitchen wastes . Another advantage of the desi chicken is the good taste &flavor of both eggs and meat as claimed by the common consumer. Some demerits of economicimportance (like slow growth rate, small egg & body size, poor egg production, late maturity andbroodiness) can be overcome with selective breeding as well as cross breeding with other highproducing exotic breeds . Currently, there are few local breeds like Aseel, Naked Neck and Desi (non-descriptive) which are available for the backyard poultry. This program is thus focusing onimprovement of productive and reproductive performance of backyard poultry in the country.

Research Activities

POULTRY & WILDLIFE

a.

Improving Rural/Back Yard Poultry for Poverty Alleviation and Empowering Women .

Project title:

Evolving new breed of chicken for rural areas in Pakistan".

Keeping in view the above situation, White Leghorn birds are being crossed with Rhode IslandRed, Fayoumi and Aseel breeds . The progeny are being distributed to the villagers for backyardpoultry production . Performancedata is shown in Table 5 .

Table 5. Productive performance of various crosses reared at rural poultry farms in and around Islamabad

*RIR, Rhode Island Red **WLH, White Leghorn

Achievements

Around 20,000 highly producing chicks (Figure 8) were supplied to the rural and domesticpoultry farmers. It is expected that as a result of enhanced egg and meat production, the livingstandards and health status of the rural population especially the women would increasesignificantly thereby helping to reduce poverty and increasing social status of rural women .

Income Generated : As a result of project activity chicks / adult bird and eggs worth Rs . 1 .34million were produced . The sale proceeds were deposited as income .

130

=AKISTAN AGRICULTURAL RESEARCH COUNCII

!No . Characters Desi RIR* xWLH**

Aseel x WLH Fayoumi xWLH

1 _ Sexual maturity (days) 21 j 188 224 1862 Egg count in 52 weeks of production 73 202 143 1993 Body weight at maturity (grams) 900 1100 1175 1025~4 (Viability up to 72 weeks of age (%) 90 81 80 83


"

After sale services regarding management, vaccination & disease prevention of poultry flocksthrough internet . telephone &training at the farm .

"

Each month approximately 200 rural/domestic poultry farmers are being entertained fordemand of chicks, management of theirflocks and other entrepreneurial help .

Future Thrust

"

Mass scale production of these High Producing Rural chicken was started in 2012 . We areable to increase the production up to 20,000 chicks during the year 2012 . We are planning toproduce approximately 50,000 chicks during the year 2013 .

"

Country level coordinated research project entitled "Genetic Improvement of Rural PoultryBreeds" was submitted to ALP for funding . The project has been approved by the TechnicalCommittee and hopefully will be started during the next fiscal year 2013-14

Wildlife

ANIMAL SCIENCES

More than 50 Research Internee from Veterinary Faculties of AJK, Balochistan, KPK, Punjaband Sind Universities were trained at Poultry &Wildlife Program.

Expansion of breeding / production & distribution/sale of crossbred chick to the rural poultryfarmers will continue .

Pakistan has been the land of spectacular scenery, having some of the greatest and beautifulmountain ranges, rivers, deserts and forests . Due to diversified eco-zone which has had a variety ofhabitat of different types, Pakistan used to hosting rich and diversified wildlife species . However.presently wildlife and their habitat have been adversely affected as a result of human activities . Themajor contributors are population explosion, advent of industrial and technological innovations, overgrazing by livestock, land erosion by wind and water, deforestation and indiscriminate hunting andpoaching etc .

In the mid-1800s, elephant, lion, and tiger were common in present Pakistan . Herds of Chinkara werecommon in the Cholistan, Thal and Thar Deserts and Persian gazelle in Chagai and Kharan deserts.Blackbuck could be easily seen in Cholistan . Urial roamed freely in Salt , Kala Chitta and KirtharRange, slopes of Northern Areas mountains and many areas of Balochistan . Ibex and Markhor werefound in most of the ranges of northern mountains and Balochistan and hog deer in all riverine tracts ofSindh and Punjab . Nilgai was commonly found in agricultural lands from Bahawalpur to Sialkot and sowas Kashmir stag in the Neelam Valley of Azad Jammu and Kashmir. Musk deer inhabited almost allbirch and alpine habitat areas of northern mountain ranges . Similar was the case with barking deer,goral and Sindh ibex in their respective habitats .

The continuing loss of forest habitat, with its associated fauna will have serious implications for thenation's other natural and agricultural ecosystems . Hence, there is need to suggest necessary stepsfor its conservation/management . Key to protect biological heritage of Pakistan is in the involvement oflocal people and in the support from research institutions, like PARC for conservation and sustainableuse of biodiversity. Lack of scientific data about the biology and ecology of these species from theirnatural habitat and captivity is a major constraint in their scientific management . In order to conserveand manage this natural resource, in nature and in captivity, data on biology ecology and behavior ofwildlife species is needed but it is very difficult to record the exact observations of some wildlife

species dueto their peculiar behavior /shy habits in their inaccessible natural habitat and it remains themain hurdle in their management in natural habitat as well as in captivity i .e ., in zoo and wildlife parks .

a.

Conservation and Captive Breeding for Stable Ecosystem/Balanced Environment.

Projecttitle : Studies on biology, captive breeding and other behavioral aspects ofindigenous endangered wild animalsand birds

Keeping in view above referred problems and hurdles in management and conservation of nativewildlife species a program was proposed for breeding of wild animals and birds in captivity . Data abouttheir breeding, behavior and feeding is being collected . Feed preference experiment has beeninitiated . Seasonal fodder cropping continued for animal feeding and planted fruit trees and shadeplants inside the section. We have achieved successful breeding of Hog deer (Axis porcinus) which isvulnerable according to IUCN Red list 2008 . Data on breeding behavior and growth characteristics etcis projected briefly in Table 6 .Table 6. Breeding behavior and growth characteristics of Hog Deer in captivity at NARC

Achievements

Future Thrust

Construction of 5 deer enclosures, 8 bird cages. one bird aviary, office/store building iscompleted

Developed permanent pasture fields and plantation inside wildlife research section for feedingneeds and shades.

Successful breeding results of Hog deer and Peafowl achieved

" Maintain already established wildlife conservation/breeding research facilities at NARC(Figure 9-11) .

"

Transfer/sharing of technology to researchers, students, wildlife farmers, breeders and wildlifemanagement and conservation related organizations & NGOs .

132

PAKISTAN AGRICULTURAL RESEARCH COUNCIL

Sexual Activity in Male : 09-10 monthsCaptivity : l s ` sign Female : 10-11 monthsticxual & plating in Sexual hchti~ for ~ca, obser~ cd froth Aril to October.Captivity Male buck was more active during the months ofApril & May .

Most mating was observed after the onset ofspring and before extremehumid hot climate .Copulation observed in morning and this observation was recorded twice atour vicinity.

Gestation Period Black Buck, 5 Months Hog Deer : 8 MonthsFawning Season Black Buck : March, September . llog Deer : February-March, November-Dec .Reproductive No reproductive disorder observed in hog deer femaleDisorders _Dy~tocia s% a> obscr~ ed in one female black buck twice : one case was

handled stjcccssfully while last one was roved fatal for both dam & fetus .Antler Cycle of Ho Deer

Age Year I Year ? Y"ear3Antlers %weight (year-t\ ise) 1?00 gram bout 800 gram both

_850 gram both

Length (cm) Antler-1 Antler-2 Antler-1 Antler-2 Antler-1 Antler-240cm 39cm 39ctn 38cm 42cm 43cm

Antler CyclePeriod (peak) of casting 6 monthsSpan of antler growth 6 monthsVelvet stripping July-Aug

" Collaboration for joint research with Wildlife Management Department UAAR, ZoologyDepartment Punjab University, Wildlife Department, University of Veterinary and AnimalSciences, Lahore, WWF Pakistan, Punjab Wildlife Department, Environmental DepartmentCDA Director, Islamabad Zoo, Lohi beer Safari, Behria Zoo .

Figure 10:Highiy productive cross bred chicks produced at Poultry & Wildlife Program, NARC

Figure 11 : Herd of Hog Deer at Wildlife Section

Figure 12 : Urial (Male) at Wildlife Section

ANIMAL SCIENCES

Figure 13: Black Buck Deer (Male) at Wildlife Section

Figure 14 : Hog Deer (Male) at Wildlife Section

ANIMAL BIOTECHNOLOGYProductive traits of milk animals, both qualitative and quantitative, are primarily determined by theirgenetic make up . In this context several genes have been identified and this information, obtainedfrom DNA loci affecting economic traits, is helping in better selection of livestock breeds . In the milkprogeny testing programs of bulls we have to wait for seven to eight years before we can select thebulls on the basis of performance of their female progeny. If we can develop some DNA test that canhelp us to identify animals having good productive genes as soon as it is born then it will save bothtime and rearing cost of productive farm animals.

Dairy cattle breeding programs in Pakistan involve utilization of imported/exotic semen from elite bullswith high genetic merit for the dissemination of alleles associated with positive economic traits, suchas increased milk yield . Introduction and dissemination of such traits is beneficial for animalproduction but likelihood of simultaneous incorporation of defective genes is increased which isundesirable . Underpresent trend of short term livestock improvement culture, Holstein-Friesian breedis widely disseminated throughout the country, with regional populations genetically linked because ofexotic semen, embryos and live animals imported from abroad . Therefore, defective alleles occurringwithin widely-used breeding lines are likely to be present in the Pakistan Holstein-Friesian andcrossbred population thereto .

Therefore, current research activities at NIGAB are focusing on identification of animals that havegreater production potential and are free from genetic anomalies . Similarly, molecularcharacterization of pathogens of food animal origin and identification and development ofimmunogenic proteins required for diagnostics andvaccine production is also actively persuaded .

Research Activities

a.

GeneticImprovement of Production Traits in Indigenous Livestock Breeds


Experiment :

Molecular analysis of genes affecting milk production traits and diseaseresistance in food animals.

Bovine and ovine genomes are being analyzed internationally for economic traits . Modern cattlebreeding strategies involve identification of milk, meat & disease resistance genes and its variantsthat can be incorporated into selection programs . In this regard several genes have beenidentified . More than 26 genes have been mapped and characterized for milk production traits andmost important include DGAT1, CSN1 S1, CSN1 S3, PRL, LEP STAT5A, BoLA-DRB3 etc. Theobjective of NIGAB is to undertake research studies focusing on identification of genotypes withgreater potential production using molecular genetic approaches .

"

Gene polymorphism for DGAT I, rc-CN and PRL genes related to milk fat and protein in Nili-Ravi buffalo, Sahiwal and Achai cattle has been analyzed . The results indicated that theanimals of Nili-Ravi buffaloes are homozygous for DGAT I (K allele), K-CN (B-allele) andPRL (G-allele) superior for milk production traits . (Figure-13)

"

Standardized PCR test to analyze defensin gene polymorphism (related to disease resistance)in Nili-Ravi buffalo.

"

Analyzed 500 Nili-Ravi buffalo samples collected for FMDV detection and their persistenceusing Real Time PCR.

b.

Genetic Status of Pakistan Cattle Population for Inherited Disorders

ANIMAL SCIENCES

Experiment :

Genetic testing for inherited disorders in Pakistani Bos indicus, Bos taurusandtheir crossbreed.

Inherited disorders are of major importance in Holstein Frisian cattle a breed that now dominatesthe world dairy industry due to high milk production . The important inherited disorder BLAD wasdisseminated globally in 1992 by this breed and reported in Pakistani Holstein Friesians in 2008and 2012 . However a range of inherited disorders recognized internationally are still in need toexplore in this breed in Pakistan . Therefore, it was deemed important to initiate inherited diseasedetection by DNA testing in Holstein Friesian and its crossbred population in Pakistan tostrengthen the National breed improvement programs . (Figure-14)


"

PCR Optimization for the amplification of gene encoding bovine CD18 and PCR amplificationof the target genes responsible for the Citrullinaemia is achieved . Similarly amplification ofselected regions of UMPS and SLC39A4 genescontaining target mutations is achieved .

"

Optimization of a PCR-RFLP based method for the genotyping of BLAD, citrullinaemia,deficiency of uridine monophosphate synthetase and bovine hereditary zinc deficiency isachieved .

"

Screening for the citrullinaemia and BLAD is in process . Screened 200 samples including AIbulls and seen some mutations trying to confirm through sequencing .

c .

Development of DNA Based Diagnostics

Experiment :

Standardization of DNA based diagnostic test for rapid detection of foot andmouth disease virus (FMDV) under laboratory andfield conditions .

Foot and mouth disease (FMD) usually results in severe economic losses and impact on bothnational and international trade within the livestock and animal products . Conventional moleculartechniques for the detection of FMDV are expensive and time consuming so a novel nucleic acidamplification method, termed reverse transcription loop-mediated isothermal amplification (RTLAMP) is being developed forFMD diagnosis and serotype differentiation .


"

Established basic facilities for Avian and Bovine cell line maintenance. These include LFBK,BHK-21, Vero, QT-35, CEL, CEK etc.

Viruses like FMD, IBD, IBV. EDS, ILT and Avian Adeno virus 4 and 8 have been isolated on celllines .

"

Standardized Lamp PCR for FMDvirus detection .

"

ForFMD persistence studies . 100 samples were tested out of these 11 found positive . Resultsshowed apparently healthy animals carry FMDV

d .

Recombinant Protein for Therapeutics and Vaccine Development


Training/Extension/Other Activities

Research Publications

Experiment : Development of cloned vaccines against avian adenovirus serotype-4 inpoultry.

Fowl adenovirus infections are widespread inmost of the poultry raising countries of the world. Fowladenoviruses belonging to group I have been associated with a number of clinical conditions likeeffect on egg production and Hydropericardium syndrome which is an emerging disease of broilersand breeders resulting in heavy economic losses . Autogeneous formalin inactivated vaccinesprepared from infected liver homogenate have been used against HPS but they are unable tocontrol the disease So present study was designed to develop a cloned vaccine against HPS

"

PCR has been successfully standardized for detection of AAV-4 from 30 different samples.20 were found positive for AAV-4 .

" Cloning and expression analysis of hypervariable region of hexon was achieved andconfirmed by PCR and sequencing .

"

Recombinant protein was expressed in BL21 cells and purified by GSTtagged columns usingGST buffer kit .

ISHAQ, R ., SULEMAN, M., RIAZ, M . N ., YOUSAF M., SHAH, A. and GHAFOOR, A. (2013),Prolactin gene polymorphism in Nili-Ravi buffaloes in relation to Sahiwal and Achai Cattle .International Journal of Dairy Technology, 66 : 2024 . doi : 10 .1111/j .1471-0307.2012 .00875 .x

"

Suleman, M, S . U. Khan, M . N . Riaz, M . Yousaf, A. Shah, R . Ishaq & A. Ghaffoor. (2012) .Calpastatin (CAST) Gene polymorphism in Kajli, Lohi and Thalli sheep breeds . Afri . J . Biotec .11(47),10655-10660 .

"

S. U. Khan . M . N . Riaz & A . Ghaffar (2012) . Calpastatin (CAST) Gene polymorphism and itsassociation with daily weight gain in Balkhi and Kajli sheep and Beetal goat breeds .

Pak . J .Zoology, 44 (2), 377-382 .

Workshops/Trainings/Conferences

" Attended two-days workshop on "Control of Dengue virus vector using advancedtechnologies" organized by NIBGE, Faisalabad on 31 "January -I" February, 2013 .

"

Attended a three-week training on "Real Time PCR based detection of FMDW organized byARD-USDA at AHP ASI, NARC in December, 2012 .

"

Participated in SAARC Regional Conference on New Frontiers in Agricultural Genomics andBiotechnology, Organized by NIGAB, HEC & MNFS&R, Islamabad on June 5-7, 2012 .

Participated in One Health Workshop 2 : Draft Programme (v3), Serena Hotel, Islamabad,Pakistan 10-12July, 2012, organized by Massey University, NewZealand .

ANIMAL SCIENCES

" Participated in the conference on Nanotechnology-Its application in Animal Health &Production at UVAS, Lahore, 16-17 April, 2012 organized by UVAS and HEC .

Conference abstracts

"

Comparative pathogenicity of cell propagated Avian adenovirus-4 and development of cellculture based vaccine. Asma Jabeen, Khalid Naeem, Tahira Kamal and S.M . Saqlan Naqvi .Society of General Microbiology Conference Spring March 2013 Manchester, UK.

"

Development of cell culture based live vaccine of AAV-4 and comparative efficacy withcommercial vaccine . Asma Jabeen, Khalid Naeem, Tahira Kamal and S.M . Saqlan Naqvi . 6`hAnnual Cell Culture Technology event Recent Advances, Future Prospects March 2013 RoyalCollege of Pathologists, London .

"

Tahira Kamal, K. Khalid Naeem. Amir bin Zahur, Ghulam Muhammad Ali, Naeem Riaz, FoziaNasreen, Aman Ullah and Asma Latif. (2013) . Comparison for the sensitivity of BHK-21 andBovine Kidney Cell Line for the Isolation of FMDviruses . Presented at 9th International BiennialConference of Pakistan Society for Micvrobiology, January28-31, 2013 . Karachi-Pakistan .

"

Evidence of foot and mouth disease virus in apparently healthy buffaloes in abattoirs ofIslamabad Pakistan . M . T Naveed, U. Farooq, N . Siddique, TKamal, A.B.Zahoor,A.Ullah,K.Naeem, S . Raza, F Rasheed and A. Latif . 9th International Biennial Conference of PakistanSociety for Microbiology . Karachi, Pakistan.pp .38, Jan 28-31,2013 .

" M. N . Riaz, S . U. Khan, F. Nasreen, M . Suleman and J . A . Qureshi . (2012) . K232Apolymorphism in Diacylglycerol Acyl transferase I gene in Nili-Ravi buffalo, Sahiwal & AchaiCattle . SAARC Regional Conference on 'New Frontiers in Agricultural Genomics andBiotechnology June 5-7 .

"

Fozia Nasreen, Naveed A Malik, Muhammad N . Riaz, Imke Tammen and J . A . Qureshi . (2012) .Screening for Citrullinaemia, DUMPS and Bovine hereditary zinc deficiency in Pakistani Bosindicus, Bos taurus and Bos indicus x Bos taurus cattle . SAARC Regional Conference on'New Frontiers in Agricultural Genomics and Biotechnology' June 5-7 .

"

Khalid Naeem, Naila Siddique, M. Naeem Riaz, Umer Farooq and M . Athar Abbas . (2012) .National research focus in Animal Health Biotechnology. SAARC Regional Conference on'New Frontiers in Agricultural Genomics and Biotechnology' June 5-7 .

"

Isolation Cloning and Sequencing of Hexon gene of AAV-4 aAsma Jabeen, bKhalid Naeem,Dure shahwar, aS . M . Saqlan Naqvi SAARC Regional Conference on New Frontiers inAgricultural Genomics and Biotechnology" hosted by NIGABat Marriott Hotel Islamabad fromJune 5-7. 2012 .

Future Thrust

Standardization of PCRforthe amplification of defensin gene and to study DNApolymorphismrelated to disease resistance in cattle including Sahiwal, Holstein Frisian and its crosses.

"

Continue analysis of prolactin gene polymorphism in Holstein Frisian and effort will be made tostudy association with the milk production traits .

138

"

Screening work, for the diagnosis of inherited disorders (BLAD, Citrullinaemia, DUMPS andBHZD) from Punjab area will be performed during the year 2013-2014 .

" A proposal is submitted for research funding as Co-PI entitled "Identification andAuthentication of Market Meat and Meat Products by Biological & Molecular approaches" toRADP On receiving its funding the molecular work will be initiated .

"

Continue work on development of DNA based diagnostics .

"

Cloning and expression of avian adenovirus serotype-4 in poultry.

"

Isolation and molecular characterization of field strains of avian viruses .

58bp

M1,1 .a4e0-svInu11u14IsIs11 .1

" ~Ir

x»411 .~ gar

-"~rlwW"ri+rri+r~ss . .

Prolactin gene polymorphism in

Chicken prolactin gene polymorphismBuffalo & Cattle breeds

Figure 15 : Phylogenetic tree analysis (Fasciola spp) of FG ITS-2 constructed using Bayesian methods indicatesthat FG ITS2 found in Pakistan is unique from the FG ITS2 found at other places .

ITGB2 gene by Rl primers

Amplification by Sl primersM

M

1

2

3

4

5 -Ve

M

C.383A>G, p.D128G (Heterozygous)

c c = 1 .. _ R c c i = r -

111 ; I Illlbp ladder

BLAD

G

~i iiiiiiiiiiior" "--

M d- -1- -1+ a+ We -1- -I-

DNA sequence chromatogram

Taq I endonuclease restriction analysis

¢° 619bp

Figure 16: Genetic analysis of Bovine leukocyte adhesion deficiency in HF and its crossbred

AQUACULTURE & FISHERIES

ANIMAL SCIENCES

Aquaculture is currently one of the fastest growing food production systems in the world withproduction level increasing at an average rate of 11 percent per year over the last decade . In Pakistantoo, the major potential for increase in fish production lies in inland aquaculture . Therefore, there is anincreased interest in the development of aquaculture production not only for the food security but alsofor larger economic benefits . Inland fish culture is currently characterized by low productivity.Therefore, there is need to increase per unit area production from fish ponds on sustainable basis .This can be achieved by introduction of fish with higher genetic growth potential, provision ofnecessary nutrition and effective disease control measures . The main objective of fisheries research isto promote aquaculture and fisheries productivity by increasing per unit area production throughintensive fish culture, introduction of high value fish to increase farmers' income and reduce poverty.Further to that focus on introduction of new and emerging ideas of income generation such asdevelopment of ornamental fish science and technology that has substantial local and overseasmarkets .

Research Activities

a.

Ornamental Fish Science and Technology

Project Title :

Culture and breeding of ornamental fishes (goldfishes, Koi carp and guppies)and feed development for their different developmental stages

Experiment 1 : Culturing ornamental fishes and comparing dietary effect .

Ornamental fish is gaining popularity in the country and hence there is a growing need to workout thepossibilities of its farming within the country . For this purpose, a study has been carried out on somefresh water ornamental fishes like goldfish (Carassius spp .), guppies (Peocillia reticulata) and Koi carp(Cyprinus carpio var. Koi) . These fishes are selected because of their popularity among the people aswell as culturists . The success of ornamental fish culture in the country will not only meet the localdemand but will also provide the opportunity to raise foreign exchange for country through it export inthe regions where there is high demand . In fish culture diet is major economic factor determiningprofitability. In this context two experimental diets, containing 25 and 30 percent crude protein (CP),were compared with imported pelleted diet (37% CP) as a control .

Salient Achievement

Fingerling of Double tail (Carassius auratus auratus), Shubinkin (C . auratus), Local Koi Carp (orangeand white) (Cyprinus carpio) and Hi fin Koi Carp (C . carpio) from localy bred stock has been stocked ineathern ponds for the brood stock development having average initial weight of 10.90 gm, 11 .50 gm,11 .25 gm and 10.25 gm respectively.

All the species showed better growth fed on diet containing 30 percent CP level and can be used as areplacement of imported pelleted diet .

Experiment 2: Natural breeding of ornamental fish

Brooders of Shubunkin (38 pairs), Double Tail (22 pairs Double Tail (20 pairs) and High fin Koi carp (29pairs) has been stocked in earthen ponds, raceways and circular fiber glass tanks for breeding trialsduring November 2012.

Breeding season of ornamental fishes has been determine such Double tail (Carassius auratusauratus) breeds in December, Shubinkin (C . auratus) breeds in January, Local Koi Carp (orange andwhite) (Cyprinus carpio) and Hi fin Koi Carp (C . carpio) breeds in February and March.

Breeding of Double tail (Carassius auratus auratus), Shubinkin (C . auratus), Local Koi Carp (orangeand white) (Cyprinus carpio), Hi fin Koi Carp (C . carpio), Mollies and Guppies has been achievedsuccessfully.

Experiment 3 :

Induced breeding of ornamental fish .

Induced breeding ensures the availability of seed during the whole breeding season. In this regardssuccessful breeding trials of ornamental fish were attempt on Local Koi Carp (orange and white),Shubinkin (C . auratus) and Double tail (Carassius auratus auratus) during April 2013 .

Experiment 4: Diet development as colorenhancer.

Three different diets supplemented with 1) spirulina (blue green alga), 2) red beet root and 3) carrotwere compared with locally developed diet (30 % CP) for color enhancement and augmentedaesthetic appeal .

Diets supplemented with red beet root were found more effective as color enhancer whereas mixtureof above three diets showed further better result .


Exhibitions of ornamental fish technology at FAO Kisan Mela at NARC, FAO World food Day at NARCand Autumn and Spring Festival at Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi for publicawareness and information on ornamental fish science and technology for economic benefits .

Germplasm of Shubunkin, Double Tail, Koi carp (orange and white) and High fin Koi carp has beentransplanted to different farmers, Directorate of Fisheries Khyber Pakhtunkhwa, Peshawar, Directorateof Fisheries, Gilgit-Baltistan and Punjab Fisheries Department, Lahore .

Culture and breeding technology of ornamental fish (Shubunkin and High fin Koi carp) has beentransferred to 5 progressive farmers of Potohar Region (Jand, district Attock) and successful breedinghas been achieved at their door step .

An amount of Rs 500,000/- has been generated through ornamental fish sale proceeds .

Future Thrust

Countrywide dissemination of culture and breeding technology of ornamental fishes (Figure 15-22) .

Breeding trials will be conducted on two new species, angel fish and sucker fish .

b.

Increasing Productivity Through Intensification of Fish Culture.

ProjectTitle : Intensification of fish culture to increase per unit area production in farmponds usingdifferent management inputs

This project has been designed to increase the fish production through intensification of fishculture and timely inputs utilization such as fertilizer and artificial diets . Judicious use of such

Figure 23: High fin Koi carp

Figure 24: Koi Carp white(Cyprinus carpio) fry

(Cyprinus carpio) Fry

ANIMAL SCIENCES

inputs and adoption of the intensive fish culture technology can easily enhance the fish productionfrom present level of about 1000-1200 kg/acre to about 1800-2500 kg/acre per year as it has beenalready demonstrated in other Asian countries such as (India, Bangladesh and China) . Therefore .this project activity was undertaken to prove and verify intensive fish culture technology bystocking participating farms in all three provinces with three indigenous or major carp, namelyrohu (Labeo rohita), mrigal (Cirrhinus mrigala) and Catla (Catla catla) and three exotic Chinesecarps i .e ., grass carp (Ctenopharyngodon idella), silver carp (Hypophthalmichthys molitrix) andcommon carp (Cyprinus carpio) .

Figure 17: Brooders of Shubinkin

Figure 18: Brooder of Double tail

Figure 19 : Brooders of Local koi Carp(Carassius auratus)

(Carassius auratus)

(Cyprinus carpio)

Figure 20: Brooders of Hi fin koi carp Figure 21 : Shubunkin

Figure 22: Double Tail(Cyprinus carpio)

(Carassius auratus Fry

(Oranda) (Carassiusa auratus) Fry

f

Salient Achievement


Future Thrust

The project has been initiated recently w.e .f . 01-10-2012 in four province of country and inNARC . The coordination has been developed between four provinces and PARC.

"

Survey was conducted to identify the progressive fish farmers' at Ali Pur Chattha, Gujranwaladistrict for the selection of fish farms . A farm was selected at Jamkay Chattha, Gujranwala forintensive fish culture.

"

Thefish were stocked at farmers' field at different stocking density i .e . 1500 fish / acre . 1800fish/ acre and 2100 fish/acre and was supplement with 20% CP artificial diet . The averageweight gain aftertwo months is 120gm.

"

Artificial diet developed for all developmental stages of carps at Aquaculture and FisheriesProgram and transferred to farmers in Intensive culture project

"

Study growth of carps fed on different supplementary fish feeds and testing of diets indifferentagro-ecological zones .

Evaluate varying stocking densities of carps.

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