ANIMAL SCIENCES
ANIMAL SCIENCES
ANIMAL SCIENCES
The Animal Sciences Division (ASD) of PARC is cognizant of the importance of livestock sector in
national economy and efforts are underway to tap the potential and opportunity's offered by the sector
for enhancing national food security, economic uplift, rural development, poverty alleviation,
employment generation and foreign exchange earnings . The main research focus of ASD has been
control of infectors diseases of livestock and poultry (FMD, PPR, HS, AI, ND), improving feeding
management strategies, increasing reproductively efficiency and boosting small ruminants
productivity. The ASD research activities are undertaken mainly at Animal Sciences Institute. NARC .
ANIMAL HEALTH
Livestock industry plays an important role in the economy of Pakistan . However, prevalence of
infectious diseases poses a major threat to the livestock productivity. The research focus of the Animal
Health Program at Animal Sciences Institute (NARC) is to study the emerging & re-emerging diseases
of livestock through use of molecular and epidemiological tools, improved diagnosis and devising
control strategies based upon the local epidemiology of the pathogens . During the period under
report, most of the research activities aimed at studying the pathogenesis & pathogen
characterization, 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 control
measures and surveillance of selected zoonotic diseases (Brucellosis and Crimean Congo
hemorrhagic fever) . This Program is also serving as hub for activities related to the Department of
Animal Genomics and Biotechnology under the degree awarding institution of PIASA (PARC Institute
for Advanced Studies in Agriculture) at NARC .
Research Activities
a. Control and Management Strategies Against Major Trans-boundary Animal Diseases
Project 1 : Carrier potential of small ruminants in the persistence and transmission of peste
des petits ruminants (PPR) virus.
Following acute phase of the disease, Peste des Petits Ruminants (PPR) virus shedding has been
reported for 11 weeks post recovery and in some animals even up to 12 weeks . It has also been
reported that animals infected by PPR virus are incubatory carriers and virus shedding was
detected 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 at
the high risk areas like weekly livestock markets and communal grazing pastures. Such
asymptomatic carriers are threat to livestock farmers and may be responsible for trans-boundary
events because the disease control measures cannot be instituted . A study was conducted to
ascertain the persistent shedding of PPR virus in the animals that experienced PPR and recovered
under field conditions . Longitudinal data were collected by following five outbreaks where the
animals were sampled after 5, 10, 15 and 18 weeks post recovery. PPR virus shedding was
detected 15 weeks post recovery. This evidence was supported by RTPCR, Efforts are underway
to recover the PPR virus from fecal material of such animals . Furthermore, all the samples will be
examined by rRTPCR to quantify the viral load .
Salient Achievements
" Hem-agglutination (HA) assay for the detection of PPR virus from persistently infected
goats was optimized .
"
One step PPR virus specific RTPCR, using F and N genes was standardized .
" Evidence of persistent PPRV shedding by recovered goats and sheep was recorded 15
weeks PPR post outbreak . This evidence was supported by HA test and PPRV F-gene
specific RTPCR .
Future Thrust
" Sampling will continue to be carried out in high risk areas (livestock markets, pastures for
communal grazing) to strengthen the evidence of PPR persistence .
"
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 of
peste des petits ruminants (PPR) in Pakistan
Pakistan is second to India in small ruminants' production in South Asia. The small ruminants are
usually kept by low income landless villagers to whom this activity provides the main source of
income . These rank second to poultry in the livestock species ladder while prioritizing the species
kept by the poor. The cost of a national outbreak of PPR was estimated to be worth Rs . 20.53 billion
annually under an ALP funded project . The parameters quantified included losses due to
mortalities (Rs . 11 .6 billion), meat loss (Rs. 7.13 billion), milk loss (Rs . 0.6 billion) and cost of
treatment (Rs . 1 .2 billion) . The magnitude of economic losses due to PPR in the country warrant
measures to be taken to control this menace . This project was, therefore, initiated with the
objective to improve the socioeconomic conditions of small ruminants producers and achieving
food security through progressive control of PPR in Pakistan .
Salient Achievements
Two stake holders' workshop were successfully planned and organized at Azad Jammu and
Kashmir (AJK) Coordinated Unit in Bhimber district (sub-district Barnala) .
Eight suspected PPR outbreaks were investigated in Punjab and AJK . The outbreaks were
confirmed by HA test, Ic-ELISA, cELISAand RT PCR .
Detection of PPR virus from the samples collected on filter paper by RTPCR was optimized
without RNA extraction .
Future Thrust
"
Stake holders' workshop will be organized in all provincial and AJK Coordinating Units .
"
PPR vaccination will be conducted in target tehsils .
"
Outbreaks will be investigated throughout the country.
"
Studies on the development of thermo-stable PPR vaccine will be conducted .
106
PAKISTAN AGRICULTURAL RESEARCH COUNCIL
ANIMAL SCIENCES
Project 3: Improving the diagnostic assay for the control of warble fly in Pakistan : Need and
justification
Warble fly is an economically important pest of cattle and goats in hilly, semi-hilly and sandy areas
of Pakistan . The major economic loss inflicted to livestock and leather industry by the larvae of this
fly result from perforation in the hides and skins. Degradation in meat quality, poor weight gains in
growing calves/kids and lowered milk production in lactating animals are the other economic
losses caused due to this disease. Economic losses of Rs . 20.6 and 2 .2 million due to warble fly
infestation 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 signs
during winter or serological methods. Direct clinical examination, generally results in an
underestimate of infestation level unless animals are examined regularly throughout the
emergence period . Therefore, there is a dire need to establish a reliable serological diagnostic test
for diagnosis of warble fly. The project was initiated with the objective to determine the efficacy of
ELISA for diagnosing warble fly infestation in local cattle and goats using local isolates of
Hypoderma lineatum and Przhevalskiana silenus .
Salient Achievements
"
Field blood sampling for the whole year has been completed from selected experimental
animals stationed at BLPRI, Kherimurat . In total, 1920 cattle and 1740 goats were sampled .
"
Antigen has been prepared from first stage larvae of warble fly for the use in ELISA]
Future Thrust
"
Development of ELISA for the detection of warble fly infested animals .
"
Determination of accurate timing for the treatment and effective control of warble fly
infestation .
Project 4: Epidemiology, pathogen characterization and pathogenesis of foot and mouth
disease virus (FMDV)
Foot and Mouth Disease (FMD) infection puts a serious constraint on the economy and
productivity of the livestock in Pakistan . Understanding of FMD epidemiology and
pathogenesis is crucial for FMD control efforts. The lack of epidemiological data like the
prevalence, incidence, risk factors, transmission and persistence patterns of FMD in livestock
and 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
"
Characterization of local isolates of FMDV and development of vector based vaccines .
"
Epidemiology, vaccines and control of FMD in Pakistan .
"
Epidemiology, pathogenesis and vaccine standardization for FMD in Pakistan .
"
Setting up of Epi Node at AHP (Animal Health Program) under FAO project on FMD control .
Salient Achievements
"
Established state of the art virology lab in Animal Health Program with the facilities of Indirect
10 7
Sandwich ELISA, NSP ELISA, Liquid Phase Blocking ELISA, conventional PCR, real time PCR,
VN, LFBK cell culture and sequencing. All these tests were standardized at NARC and
harmonized with PIADC .
"
Established a bio-secure animal house facility at Animal Health Program .
"
Established Federal Epidemiology Node atASI, NARC .
"
FMDV sequencing protocols were standardized and partial sequencing (VP1) of 42 selected
FMD field isolates collected during the years 2007 to 2010 and three vaccinal strains were
carried out. Variation is observed in FMDV serotype O and A, while Asia 1 is stable .
The frequency of FMD outbreaks reported from various regions of Pakistan and FMD virus
serotypes identified in these outbreaks during 2007 to 2012 . Error! Reference source not
found .FMD virus serotypes A, O and Asia 1 are identified serotypes with O being most
prevalent . Majority of these outbreaks have been reported in early to mid winter, owing
possibly to improve reporting and surveillance network.
Vaccine matching studies were carried out using Merial and Arriah vaccines . The pooled sera
obtained from Arriah vaccine reveled that serotype O has good matches, while A and Asia 1
have not good matching with circulating strains in Pakistan . This will lead to the selection of
appropriate vaccine strain for FMDV control in Pakistan and developing regional FMDV
vaccination programs .
Results of the persistence studies indicated that the Asian Buffaloes are carrier of FMDV Virus
shedding was observed in oro-pharyngeal (OP) fluids and milk collected from buffaloes
indicating the possible role of apparently healthy buffaloes in spread of FMD.
"
Over 600 outbreaks were reported from district Karachi only, suggesting a highly endemic
area .
Future Thrust
"
The project has been successfully completed and the remaining planned activates will be
covered 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 be
conducted at a bio-secure facility, which is being constructed at animal health program,
NARC .
"
Federal Epi-node on FMD will be made functional for data management and advising in future .
b . Epidemiology and Diagnosis of Zoonotic Diseases One Health Approach
Project 1 : Establishment of Field Epidemiology Laboratory Training Program (FELTP) for
joint-human/animal disease investigation against Brucellosis, Crimean Congo
hemorrhagicfever and avian influenza
In the past few years, emergent disease episodes have increased globally . The list of important
emergent diseases is impressive indeed and, given what we know about disease ecology, it will
only continue to grow . Nearly all of these emergent disease episodes have involved zoonotic
ANIMAL SCIENCES
infectious agents : the agents transmissible from animals and wild life to humans. The countries
that have a national plan for the control of specific zoonotic diseases based on the available
resources and strategies are able to eradicate many such diseases . In spite of the progress
achieved, the diseases like tuberculosis, Crimean Congo hemorrhagic fever (CCHF),
Leishmaniasis, brucellosis and rabies still constitute a threat to human health and welfare
particularly in Asian region . The same is true for Pakistan . Therefore, there is a dire need in Pakistan
to understand the burden and epidemiology of zoonotic diseases in different regions of the
country in livestock and poultry among high-risk groups of the population. Besides this, the risk
factor associated with these diseases needs to be studied so as to plan and implement
appropriate interventions within the livestock sector, animal-human interface and the human
population . The priority zoonotic diseases of animals selected for this study currently include
Brucellosis, CCHF and Avian influenza.
Salient Achievements
" A countrywide network has been established for the surveillance of brucellosis and CCHF In
this regard 19 provincial surveillance sites (PSS) have been set up.
" A total of 1399 serum samples of livestock origin were received from these PSS out of which 342
were found positive for brucellosis by Rose Bengal Plate Test .
B . millitensis isolated from afield outbreak in a sheep and goat flock (first isolate in Pakistan) .
" 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 at
Animal Health Program, NARC, Islamabad .
Future Thrust
To expand and strengthen existing surveillance network for brucellosis and CCHF
Capacity building of field staff and farmers on epidemiology, disease reporting and outbreak
investigation and awareness of farmers especially women about zoonoses .
Joint vet-med meetings for collaborative work on human-animal interface to achieve One
Health Goal .
Training
" Training of two field staff were arranged in which 27 provincial field veterinarians were trained
in the area of surveillance, awareness and reporting and data collection of selected zoonotic
diseases .
ANIMAL NUTRITION
Inadequate feed resources and imbalanced conventional feeding practices are major causes of low
livestock productivity in Pakistan . Major feed resources are crop residues, green fodder, and
concentrates . The availability of green fodder is limited in peri-urban commercial herds forcing these
farmers to rely on crop residues and concentrates . The concentrates including cereal grains, industrial
by products, oil seed cakes/meals, and molasses are very expensive and used mostly in small
quantities, resulting in inadequate nutrient provision to the animals . In this scenario, the Animal Nutrition
Program at Animal Sciences Institute NARC focuses research primarily on the development of
appropriate and cost effective feeding systems/tech-nologies for farmers and commercial
entrepreneurs . In this regard, the priority areas include efficient utilization of crop residues and
agroindustrial by-products, and exploration of the latest developments in feeds and feeding. Having a
well-equipped laboratory on nutritional analyses and highly trained scientific personnel, this Program is
also striving for human resource development regarding livestock feeding. The laboratory facility and
expertise are also extended to provincial research institutes, farmers and small-medium feed industry.
Research Activities
1.
Effect of Stair-step Feeding Scheme on Weight Gain, Nutritional Input, Feed Efficiency and
Reproductive Performance of Buffalo and Sahiwal Heifers :
In stair-step nutritional regimens (SSNR), a period of growth restriction followed by a period of growth
compensation are imposed on growing heifers . Objective of SSNR is to reduce dietary energy input for
a limited time and then compensate this effect by supplying excess dietary energy to exploit the effect
of compensatory growth .
Feed cost represents about 60% of the total input costs on heifer rearing . Therefore, managing heifers
with 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 36
months (1
year more than exotic and crossbred cattle) . In previous years, the ANP already has
shown that SSNR results in reduced feeding cost (by 11%) in raising heifers from 7 to 25 months of
age, and this is without any negative impact on growth rate, time to puberty, and conception rate . In the
present 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's
dietary 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 energy
levels followed by 2 months feeding on 120% of the NRC recommended dietary energy, followed by
starting the heifers on 80% NRC recommendation again and vice versa . Results regarding Sahiwal
heifers are presented in table 1 .
'/1
Table 1 . Performance of Sahiwal heifers fed control versus stair-step nutritional regimen
Groups
Parameters
Control
I
Stair-step
-,'I re
h
W,"
Avera.
'1[ 1i"Z.zi . 196 .17
194 .17
Avera e total weight after 196 days (k)
344 .17
335 .62
Average daily weight gain (k)
0.796
0.686
Daily D ry Matter intake (kg/head)
5.728
5.500
Feed conversion ratio (kg of feed/kg wt gain)
7.196
8.017
Heifers attaining puberty (number) upto 24 months
4(67%)
5(83%)
M ature BW at first oestrus upto 24 months (%)
98 .33
95.89
Savings over control (Rs .)
2138
TI
110
.
;
°AKISTAN AGRICULTURAL RESEARCH COUNCIL
~-
ANIMAL SCIENCES
Figure 1 Sahiwal heifers in Animal Nutrition Program
In case of Nili Ravi heifers, comparatively little higher weight gain (0 .565 vs. 0.536 kg/head) and better
feed efficiency (10 .35 vs . 10 .74) was observed in heifers of control group compared to stair-step
heifers however, dry matter intake in both groups were almost same (5.85 vs . 5 .76 kg/day). No buffalo
heifers 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-ruminant
animals, such as poultry, lack significant amounts of endogenous phytase enzyme, which is required
to separate P from the phytase molecule . Because phytase from plant feedstuffs is unavailable for
absorption, the unabsorbed phytase passes through the gastrointestinal tract, elevating the amount
of P in the manure . Excess phosphorus excretion can lead to environmental problems. On the other
hand, ruminant readily use phytase because of the phytase produced by rumen microorganisms .
Need &Justification
Phosphorus availability in
cereal grains and vegetable
meals is low due to presence
of phytate . Availability of P in
these feedstuffs varies with
variety of plant and cultivation
conditions of crops . In
Pakistan, available P values of
feedstuffs have not been
determined, so the values of
other countries are being
used by poultry feed industry.
Exogenous phytase enzymes
have shown promising results
for the release of bound P in
phytate complex . Bio-efficacy
of the products needs to be
tested under local conditions
in 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 determine
bio-availability of P Six growth performance trials on broiler chicks will be conducted to determine
comparative bio-efficacy of commercial phytase enzymes.
Salient Achievements
This project is ongoing and in the first phase metabolic trials (Figure 1)on cockerels have been
completed. The analyses regarding feed and fecal samples from the first phase is undergoing . In this
upcoming stages, growth trials on broilers will be conducted .
3. Preliminary Investigations on Risk Factors in Cottonseed Cake for Dairy
Animals
The quality of cottonseed cake has been a cause of public concern during recent years. Many of the
cottonseed cake batches produced in 2011/12 were reported to be refused by cattle/buffaloes . In
some instances, an alleged contamination of CSC resulted in total feed refusal and/or mortality at
dairy farms . Present investigations were carried out as a preliminary attempt to validate the news
reports 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 million
hectares for the year 2010-11 . The corresponding cottonseed cake production was approximately 2 .3
million ton, making it the most abundantly available vegetable protein source for dairy animals . Any
contamination of cottonseed cake, the main ingredient in dairy feed, with dietary contaminants is not
only a health hazard to the dairy animals but also a risk for consumers via indirect contamination of
milk.
Salient Achievements
Six samples of cottonseed cake associated with feeding problems, along with samples not
associated with such complications were collected. Stake holders were interviewed for gathering
history and symptoms of feeding problems associated with cottonseed cake. Results of the initial
investigations are as under :
"
The main complaint of the dairy farmers regarding low quality of cottonseed cake was feed
refusal by cattle/buffalo, along with reduction in milk production . At some farms, symptoms of
toxicity alleged to be due to cottonseed cake feeding included diarrhea or respiratory
problems, which could be reversed afterwithdrawal of the cake feeding .
"
Information collected from the stake holders indicated that low quality batches of the
cottonseed cake are associated with several months of post-production storage, indicating
possibility of fungal invasion during hot and humid climatic conditions .
"
Cadmium exceeded the tolerable limits of 0.2 ppm in all the analyzed samples including
controls (0 .6-3.1 ppm in the suspected samples vs . 0.3-0 .6 ppm in the controls), while lead
and chromium at toxic levels were also detected in samples from one city.
Detailed analyses for mycotoxin contamination revealed high levels of aflatoxins and
ochratoxins in all the samples including controls. Overall, heavy contamination of the
AKISJAN AGRIC;L.IL-CUFiAL HESEARCN COUNCIL
ANIMAL SCIENCES
cottonseed cake with toxigenic Aspergillus, Penicillium, Claviceps, Fusarium,
Alternaria/Phoma, and Arthrinium spp was found . The novel finding in this regard was the
presence of tremorgenic mycotoxins in excessive quantities from Claviceps and Aspergillus
spp. i n the samples suspected of feed refusal and toxicity problems.
Training/Extension Activities
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 at
following venues :
"
Institute of Animal Nutrition & Feed Technology, University of Agriculture, Faisalabad (14th
March, 2013)
"
Research Centre for Conservation of Sahiwal Cattles (RCCSC), Jhang (25th March 2013) .
"
Livestock Production & Research Institute (LPRI), Bahadur-Nagar (26th March . 2013) .
"
Buffalo Research Institute (BRI) Pattoki (27th March 2013) .
2.
A seminar on "Preliminary investigations on cottonseed cake implicated with feed refusal and
cattle poisoning problems in Pakistan" was delivered in the 67th Conference of the Society of
Nutrition 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 popularizing
nutritionally balanced blended cattle feed and helping farmers and small-medium scale feed
millers in operating feed mills . Additionally sale of Multi-Nutrient Urea-Molasses Blocks to farmers
in the areas with fodder scarcity has been a major activity. In this regard following are the
achievements for the last year:
"
5600 bags of PARC Cattle Feed . Figure-2
"
1500 Multi-Nutrient Urea-Molasses Blocks .
"
350 sachets of Milk Booster (mineral premix) .
Figure 3. Different products produced at the Feed Technology Unit of the Animal Nutrition Program
"
30 farmers and small-medium scale feed millers from various parts of the country were given
technical assistance .
Future Thrust
Results regarding the stair-step heifer development for early and economical heifer
development will be disseminated to the farmers and other stake holders .
Laboratory analyses and further metabolic trials will be conducted regarding the project on
bioavailable phosphorus in indigenous feedstuffs in poultry.
Investigations will be continued on cottonseed cake samples associated with feeding
problems .
A research proposal on assessment of amino acid availability in locally available feedstuffs for
chickens is under process .
11 4
PAKISTAN AGRICULTURAL_ RESEARCH COUNCIL
ANIMAL SCIENCES
ANIMAL REPRODUCTION
Improvement in reproductive efficiency of buffalo and cow is the major objective of present research
activities at the Animal Reproduction Program . Buffalo and cow are both important dual purpose (milk
and meat) animals of Pakistan . Artificial insemination (AI) is the most important technique to improve
genetic potential of livestock herd in shortest possible time. First research area is to improve post-thaw
semen fertility which is especially required in buffalo as the conception rate in buffalo with frozen
thawed semen has been reported low in large scale field trials . It has been observed that improved
semen 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 with
improved management practices . Due to scattered livestock population and small holdings (1 to 7
animals per farmer), application of AI is not easy on large scale. Bringing large number of animals in
heat at one time in a particular area will make application of AI in these animals convenient . Efficacy of
this technique can be further improved by application of better management practices such as use of
male effect called biostimulation . Furthermore, this line of action will be used to overcome seasonality
of 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 buffalo
bull spermatozoa
During cryopreservation of semen up to 50% viability of sperm is lost and the main reason is the
production of toxic reactive oxygen species (ROS) . Sperm cells are exposed to oxygen and visible
light radiation during processing of freezing leading to the formation of ROS . Albumin is an
important extra cellular antioxidant minimizing the formation of ROS (OH- radical) . Objective of the
present study was to determine the effect of adding bovine serum albumin (BSA) at two different
concentrations (0.5% and 1 .0%) in tris citric acid extender on post-thaw quality of buffalo bull
spermatozoa .
Salient Achievements
"
Semen frozen in extender containing 0.5% BSAwas significantly better that that %vith 1 .00, BSA
or control in terms of post thaw acrosome integrity
"
Addition of 0.5% BSA in tris citric acid extender gave higher conception rate than control after
Artificial Insemination in buffaloes .
Experiment 2: Effect of duck egg yolk plasma in extender on post thaw quality of buffalo bull
spermatozoa
Egg yolk from chicken and ducks has been used as a component of semen freezing extenders for
most of the animal species including buffalo probably because of its easy availability. It is
hypothesized that plasma from duck egg yolk can replace the whole egg yolk in tris citric acid
extender as it has the potential to preserve the post thaw quality of buffalo bull spermatozoa . This
study 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 acid
based extender on the basis of post thaw viability of buffalo spermatozoa.
Salient Achievements:
"
The Computer Assisted Semen Analyzer (CASA) motility parameters were better in extender
containing 15% plasma from duck egg yolk although the difference was not significant
"
Whole duck egg yolk or whole chicken egg yolk can be replaced with 15% plasma from duck
egg yolk in tris citric acid based extender for cryopreservation of buffalo bull spermatozoa.
B. Improving Reproductive Efficiency of Cows and Buffaloes Through Estrus Control and
Better Management
Experiment 1 : Effect of biostimulation during estrus synchronization on conception rate in
buffalo
Reproductive efficiency is one of the primary factors affecting productivity of buffaloes . Hormones
have been used over last two decades to stimulate reproductive system in cattle . However use of
hormones without improved management practices like nutrition, housing and bio-stimulation will
never be able to stimulate animal body to achieve the desired objectives . Bio-stimulation in the
form of presence of a bull in the herd is considered beneficial to keep ovaries active and bring
animal into heat . Furthermore there are indications that biostimulation results in improved
conception rate in cattle . Present work was conducted to assess the effect of biostimulation on
fertility in buffaloes treated for oestrus synchronization with GnRH + PGF2a (ovsynch program)
with an assumption that conception rate will further improve
Salient achievements
"
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) .
Training/Extension Activities
"
Six training courses (two week each) were conducted for community workers from Punjab and
Balochistan on artificial Insemination and pregnancy diagnosis in cattle and buffalo (total
participants = 90) .
"
Imparted training to community workers from Sindh, Gilgit-Baltistan and Chitral on Modern
techniques of livestock production
Forty DVM internees from Veterinary faculties of Balochistan, Sindh . AJK, Punjab and KPK
Agricultural Universities were given practical training in the latest Animal reproduction
techniques
Four M .Phil students conducted thesis research
Future Thrust
"
11 6
Investigations will be carried out on Fertility-Related Biomarkers in Buffalo Semen to Reduce
Male-Factor Losses
-'AKISTAN AGRICULTURAL RESEARCH CCIUNCII_
ANIMAL SCIENCES
"
Effect of presence of bull will be studied during summer season in buffalo
"
Work will be conducted on fertility of Sahiwal cows after estrus synchronization at farmer level
d
m
c0
.a
d
u
C
O
V
Figure 4. Buffaloes synchronized for estrus with
ovsynch (GnRH + PGF2a)
Oestrus
synchronization
alongwith
biostimulation
Oestrus
synchronization
with no
biostimulation
Figure 5. Conception rate in buffaloes treated with
ovsynch program for estrus synchronization in the
presence 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 remains
under 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 liter
batches) which are generally inefficient and being primitive lack quality and hygiene thus posing health
hazards and draining farm economy. These processors do not have access to trainings and often find
difficulties 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 resultoriented trainings on hygienic milk production and cottage scale dairy product preparation . The
ultimate target is value addition to milk by expanding dairy product variety and thus business thereby
generating better employment opportunities for small investors both in rural and urban areas .
ANIMAL SCIENCES
Research Activities
a.
Farm Milk Quality
Experiment 1 . Evaluation of farm milk and preparation of value added dairy products
Although Pakistan stands first in milk production among Muslim World and 4th in overall world
ranking and is presently producing about twenty 47 million tones of milk annually, yet quality of the
milk and milk products is not satisfactory which is generally due to poor management techniques
being practiced by dairy farmers . Apart from milk quality, the quantitative production is also low and
in due course oftime average animal productivity with respect to milk quantity is declining . Feed and
genetics 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 respect
to our milch breeds . These are not only important as economic traits of animals but at the same time
quality 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 volume
of 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 milk
quality as it is a window to the production of individual animal in a herd to run their farm more
efficiently. Furthered to that, value added dairy products, particularly spicy cheese & yoghurt
varieties and relevant training to small-holders / internees for diffusion of knowledge were also part
of this undertaking .
Salient Achievements
"
Comparing on average content basis, buffalo milk was found richest in fat (6.0%) and milk
solids (14 .6%) while cow milk was lowest at 4.4 and 13.1 %, respectively (Tables 2 & 3) . The
averages stated met the defined respective standards but did not match real/normal field
values particularly SNF content of buffalo milk.
"
The corresponding protein values of both types of milk are somewhat within normal limits but
ash contents are significantly lower in buffalo milk which also reflected from low SNF contents
of buffalo milk. Buffalo milk is also low in lactose contents . On overall quality basis buffalo milk
was found 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
143 C
2
150 C
4
168 C
3
5
158 C
4
387 C
147 C
5
Grand Mean ± SD
Total Solids
11 .6--0 .1
12 .1±0.3
13.0±0 .4
13 .2±0.5
13 .2±0.9
15 .4±0.8
13.1±0.5
Fat
3 .3-0 .1
3 .9±0 .2
4.2±0 .4
4.4±0 .6
4.6±0 .9
5.9±0 .8
4.4±0.5
SNF
8.3±0 .1
8.2±0 .3
8.8±0 .1
8.8±0.2
8.6±0.3
9 .5±0.2
8.7±0.1
Protein
3 .0±0 .1
3 .0±0.1
3 .2±0.1
3 .2±0.1
3 .2±0.1
3 .5±0.1
3.2 ± 0.1
Lactose
4 .6±0.1
4 .5±0.2
4 .8±0.1
4 .9±0.1
4 .7±0 .2
5 .2±0 .1
4.8±0.1
Ash
0.7±0 .1
0.7±0 .1
0.7±0 .1
0.7±0 .1
0 .7±0.1
0 .8±0.2
I 0.7±0 .1
"
Animal ranking/comparison, according to milk quality, has been shown in Figures 5 and 6
which 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. The
detailed milk composition comparison helped to predict animal productivity and thus serve as
powerful tool in farm management .
"
A variety of dairy products was produced and evaluated by a taste panel . It was observed that
buffalo 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 are
extended to farmers (individually or in groups), cottage business persons, small producers or
processors and university graduates . In the year under report major focus of training was student
internships which included both class room instructions and hands-on practice on milk analysis and
product preparations ; a total 1225 man-hours on training was spent .
"
Trainings and extension services were extended to farmers (individually or in groups), to
students (as internee or class room teaching) . women farmers / activists or house wives . Some
of the trainings were organized in collaboration with some agencies such as Agriculture
Polytechnic 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 onfarm milktesting etc .
Participated in four Agri Exhibitions and displayed various dairy products for public awareness
on milk and milk product quality and to convey information about training opportunities in this
field at NARC .
Future Thrust
Studies regarding development of suitable non-gelatin based stabilizer for improving
consistency and texture of yoghurtwill be conducted
Table 3.Percent milk composition (Means ± SD) of selected cows of NARC Dairy Farm
Tag No. N
1,52 -B
2
454 -B
5
547 -B
4
183 -B
3
1461 -B
2
3 5 -B
5
2
495 -B
539 -B
3
Grand Mean ± SD
120
~-T 1 i
Total Solids
13 .8±1 .1
14 .0±1 .0
14 .3±1 .3
14 .7±1 .8
14 .9±0 .3
14 .9±0.5
15 .0±1 .3
15 .3±1 .1
14 .6±1 .1
Fat
5 .,5-1 .4
5 .4±0.7
6 .0±1 .1
6 .0±2 .2
6 .0±0.2
6 .1±0.2
6 .4±1 .1
6 .6±1 .5
6.0±1.0
SNF
8 .3±0 .2
8 .6±3 .3
8 .3±0 .2
8 .7±0 .4
8 .9±0 .1
8 .8±0 .4
8 .6±0 .2
8 .7±0 .4
8.6±0.7
Protein
3 .9±0 .1
4 .1±0 .2
3 .9±0 .1
4 .1±0 .2
4 .2±0 .1
4 .0±0 .1
4 .1±0 .1
4 .1±0 .2
4.1±0.1
Lactose
3.7-0 .1
3.8±0 .2
3.7±0 .1
3.9±0 .2
4.0±0 .1
4.1±0 .1
3.8±0 .1
3.9±0 .2
3.8±0.1
Ash
-0.6-0 .1
0.6±0 .1
0.6±0 .1
0.6±0 .2
0.6±0 .1
0.6±0 .1
0.6±0 .2
0.6±0 .1
0.6±0.1
ANIMAL SCIENCES
16 .0
14 .0
12 .0
10 .0
6.0
6,0
4.0
2,0
0.0
539-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
E SNF 0 Protein
ELactose
Ash
Figure 7: Percent composition of fresh milk of selected
buffaloes at NARC Dairy Farm
149-c
168-c
Total Solids " Fat
150-c
147-c
387-c
158-c
Tag No . of cows
0 SNF
Protein
ELactose
Ash
Figure 8: Percent composition of fresh milk of selected
cows at NARC Dairy Farm
.
Research work on "identification and authentication of market meat and meat
products using molecular approaches" has been planned .
"
Continue participation and dissemination of research-based information and
technology through exhibitions, publication and trainings etc.
0
Continue offering trainings
SMALL RUMINANTS
Pakistan is rich in livestock resources especially ruminants . Currently there are about 28.4 million
sheep with 28 well know breeds and 63 .4 million goats with 25 well known breeds in Pakistan . Small
Ruminants are producing about 0.629 million tons of mutton and 0.775 million tons of milk annually. In
addition to supply of proteins of high biological value, these animals are also contributing significantly
in the socio-economic aspects of national economy. Small Ruminants play an important role in the
livelihood of landless and small farmers as well as in the overall national economy. Small Ruminant
Research Program (SRRP), Animal Sciences Institute, NARC strives for the improvement of animal
food products (e .g ., mutton and milk) through research endeavors on small ruminant production. The
Program has the facilities of rearing and experimentation of small ruminants . In countries with advance
wool producing and processing industries objective measurement of the physical characteristics of
wool is an important tool to determine price and predict processing performance of a particular lot of
wool, similar opportunities are available at the Wool Testing Service of the Small Ruminant Research
Program, ASI, NARC . Whether, one needs objective information to produce good quality yarn, check
the quality of imported or locally produced wool or wants to carry out research on wool quality, the
Wool Testing Service, NARC may be helpful .
Research Activities
a.
Exploiting Fattening Potential to Improve Mutton Production
Project 1 . Study on production potential of different sheep and goats for mutton production
under high input system"
Mutton production gap between the conventional farming system and commercial farming
system is significantly wide . Under traditional farming system of Pakistan, small ruminant's weight
gain is often low, however daily weight gain under balanced feeding and management program
can be improved significantly . Improved system of production like energy and protein
supplementation increases the production performance, survivability and growth rate in small
ruminants . Therefore need was felt to investigate the potential of sheep and goat breeds for
commercial meat production on high input system rather than extensive system . This study was
therefore 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, were
conducted and results are shown in table 4 .
Table 4 . Carcass evaluation of male Thalli sheep
Parameters
Group 1
(Concept rate
+ grazing)
No. of animals slaughtered
(100 % VCR
+ grazing)
Group 3
(Concentrate ad
libitune + grazing)
3
3
Average live weight before slaughter (kg) 68.8±8.31
49 .7+2.57
56.0±8 .26
Average live weight after slaughter (kg)
66.3±8.31
48 .0+1 .80
53 .8±8.01
Carcass weight (kg)
34 .4±2.98
24.5±1 .24
27 .2±2.93
Dressing (%)
50.0
49 .2
48 .5
Blood (kg)
2.5±0.00
1 .7±0.76
2 .2-0.29
122
3
Group 2
-'AKISTAN AGRICULTURAL RESEARCH
ANIMAL SCIENCES
Parameters
Group I
(Concent rate
+ grazing)
Group 2
(100 % NCR
+ grazing)
Group 3
(Concentrate ad
libitum + grazing)
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
Salient Achievements
"
On the basis of above referred successful trials, the Program in collaboration with Range
Research Program . fattened fifty male goats purchased by PATCO for sale at the occasion of
Eid-ul-Azha .
"
The PATCO earned hundred thousand rupees as profit from the sale of these animals
Training/Extension Activities
Meetings
The Program Leader, SRRP attended the meeting of Agricultural Innovation Project (AIP) held
in Islamabad on March 9, 2013.
"
The PI . RADP-ASD Sub-Project "Study on the production potential of different sheep and goat
breeds for mutton production under high input system" attended the Mid-Year Review Meeting
of 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 2404-2013 . They also held meetings with AIP (Livestock Component) focal persons (Program
Leader. SRRP being the focal person on small ruminants) .
The Program Leader, SRRP has been appointed as Country Coordinator (Pakistan), Asian
Australasian Dairy Goat Network (AADGN) of Malaysia .
Seminars
"
The scientists of SRRP attended a special seminar entitled 'A Conversation on Challenges
12 3
Associated with Global Food Security Focusing Animal and Agriculture" on April 11, 2013 at
the Conference Room of IABGR (PGRI), NARC delivered by Prof. Dr. Muquarrab A. Oureshi
(NIFA/USDA) .
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 .) Livestock
Management atAllama Igbal Open University, Islamabad on Sep . 27, 2012.
The DVM (Final Semester) student internees from University of Rawlakot, Azad Kashmir were
trained on small ruminant production fortwoweeks .
"
A delegate from Small Industries Wing, Government of Balochistan visited Wool Lab, NARC
on November 19, 2012 for training on wool analysis.
"
The SRRP imparted two-weeks practical training on small ruminant management and
practices to the Internees-DVM students of Sindh Agriculture University, Tandojam during
February, 2013 .
"
Program Leader (SRRP) attended the Workshop arranged by Ministry of National Food
Security & Research at NARC on February 20, 2013 .
The SRRP imparted practical trainings on small ruminant management and practices to the
different batches of Internees i .e ., DVM students of SAU, Tandojam, Pir Mehr Ali ShahUniversity of Arid Agriculture, Rawalpindi and UVAS, Lahore during March, 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 on
Biometry at Conference room of Animal Nutrition Program, ASI on March 20, 2013 .
The Program Leader (SRRP ASI) attended the consultative workshop on understanding
drivers of ecosystem change and livelihoods in the upper Indus basin on April 29-30, 2013
held in Islamabad .
"
The Program Leader, SRRP attended the training workshop on "Financial Management
System of PARC" on May 6-10,2013 at NARC.
"
The SSO SRRP gave lectures & practical trainings to the class of Livestock Management on
Animal Breeding at Livestock Research Station, ASI in April and May, 2013 .
Visits
"
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 for
guidance on developing a PC-1 on exotic dairy goat breed .
12 4
-~AKISTAN AGRICULTURAL RESEA
ANIMAL SCIENCES
"
DG (Research) KPKvisited SRRP on September 18, 2012.
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 Ruminants
Management 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 Ruminant
Research Program on January 31, 2013,
Dr. Dilpazeer, Livestock Department, Azad Jammu & Kashmir visited Small Ruminant
Research Program on February 06, 2013 .
"
Dr. Chaman Lal, Lecturer, Allama Igbal Open University visited SRRP along-with students to
witness the small ruminant research facilities at NARC on March 19, 2013 .
Forty-seven farmers visited/contacted SRRP for guidance on small ruminant management
and 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 special
reference 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) Gene
Polymorphism and its Association with Average Daily Weight Gain in Balkhi & Kajli Sheep and
Beetal Goat Breeds. Pakistan J . Zool . Vol .44 (2), pp : 377-382 .
"
Khan, M .F. and F Ashfaq, 2012 . Dairy Goats in Pakistan : Potential, Opportunities and
Challenges . In Proceedings : 1 st Asia Dairy Goat Conference, University Putra Malaysia, Kuala
Lumpur, Malaysia, 9-12 April, 2012 . pp : 60-64 .
Future Thrust
"
Improvement of local goat breeds through crossbreeding with Boer goat.
"
Studies on improvement of sheep and goat production .
POULTRY DISEASE CONTROL
Poultry sector in Pakistan is one of the fastest growing sectors of livestock segment of agriculture
based economy of the country. A steady annual growth of 10-12% has been observed since 1990 in
this sector. However, one of the limiting factors in further growth in this sector has been the intermittent
introduction of infectious diseases . Avian health is major concern for the farmers and backyard poultry
owners in the wake of high prices of poultry meat produced in Pakistan . Especially with the previous
introduction of bird flu H5N1 in this country during 2006-08, and its subsequent threat due to its
continuous circulation in other countries of the region, need for food safety and food security has
increased manifolds . Since 2003, Avian Influenza viruses of different types have been severely
affecting the growth of commercial and backyard poultry . With the recent outbreaks of highly
significant poultry diseases of zoonotic potential, enhanced diagnostic capacity at national level is
needed . Although various diagnostic kits, reagents, drugs and various vaccines are being imported
for the control of different poultry diseases, however, these drugs and vaccines are less effective
against local strains of these poultry pathogens . Keeping in view the above scenario, the National
Reference Lab for Poultry Diseases (NRLPD) at the Animal Sciences Institute of NARC has been
extending surveillance, diagnosis and research activities in this country. The NRLPD is equipped with
the state of the art equipments, highly qualified and trained manpower and strong national and
international collaborations in the field of avian disease diagnosis and research .
Research Activities
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 Avian
Health, a network of 10 labs (Figure-7) has been set up in different parts of the country in collaboration
with the provincial poultry production offices . Furthermore, due to the emergence of new strains of
highly virulent avian pathogens in the country, efforts are required to undertake relevant research in this
regard . Above all there is a strong need to undertake capacity building of the federal and provincial
institutions involved in surveillance, research and diagnosis of avian diseases . The current activities at
NRLPD are destined to achieve all above .
Salient Achievement
"
The NRLPD provided nation-wide surveillance for avian diseases and obtained data for
National AI Surveillance . 11603 samples were processed and 73 isolates of various poultry
pathogens (AIVs, H9N2, NDV, IBV, IBD, AAVs, ALV, MDV APV MG, MS, Pasteurella, E . Coli and
Salmonella) were made .
"
Production of Diagnostic reagents and their provision to public sector lab was done . The
following viruses recovered from the field were characterized and sequenced : 30 of NDV, 7 of
AIV-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 vaccines
used in commercial poultry.
"
In an experiment, differential pathogenesis and its transmission for AAV-4 was studied . Here
25 isolates were studies and RFLP- PCR was standardized for AAV The data revealed that in
ANIMAL SCIENCES
AMW
most of the outbreaks of Hydropericardium syndrome 3-4 sub-genotypes of AAV-4 were
involved 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 currently
used vaccines of poultry pathogens, revealing significant antigenic variation of the circulating
field 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 major
GP companies in this country
"
Two national trainings having 24 participants were conducted for extending surveillance
procedures for different Poultry diseases.
Two M . Phil students completed their research work and 53 internees were extended 4-6 week
training each
"
Two posters were presented in International Microbiology conference in Karachi
[MJ
- CENTRAL LAB/ NRLPD
ISLAMABAD
- PROVINCIAL LABS
O
O
0
SINDH
PUNJAB 81SD
BALOCHISTAN
NWFP & FATA
NORTHERN AREAS
_ AJK
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.
"
Study pathogenesis of new salmonellae.
"
Pathogenesis study on NDV isolates 2009-14.
b. Epidemiology and Diagnosis of Zoonotic Diseases ONE HEALTH APPROACH
Project-1 : Establishment of Field Epidemiology Laboratory Training Program (FELTP) for
joint human/animal disease investigation against avian influenza
The emergence and re-emergence of zoonosis and their potentially disastrous impact on human
health is a growing concern around the globe . The ease and speed of international travel ; the rapid
growth of international trade in poultry and poultry products due to globalization, urbanization of
populations, and dramatic changes in poultry production systems and patterns are important factors
in the spread of poultry diseases and zoonosis . Surveillance and the early diagnosis and detection of
poultry 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 in
different 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 to
be developed in future . In order to support any future surveillance activities, there is a defined need to
develop a trained veterinary workforce, reinforce laboratory capacity to test zoonotic influenza
infection samples, and influence the way the animal-human health surveillance system develops . A
successful system requires capacity at all three levels but particularly at the local and provincial level
where diseases are first identified and reported, and where response must take place.
Salient Achievements
A total of 19 provincial surveillance sites have been set up consisting of Punjab (n=3), Khyber
Pakhtunkhwa (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 AIV and 18 H9N2 AIVs were isolated .
"
14 H9N2 AIVs were sequenced for HA and NA genes and cartography experiment was carried
out to evaluate immunogenic potential, revealing that more than 3 isolates showed distinct
variation in their antigenicity, requiring their including in new vaccines .
6 meetings have been arranged under one health concept project.
Two international research papers were published and two posters were presented in
International ASM meetings .
c.
Regional Cooperation on Highly Pathogenic Emerging Diseases (HPEDs) in SAARC
Countries 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 health
services 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 are
coordinating and leading a network of national diagnostic laboratories, primarily focusing to maintain
128
PAKISTAN AGRICULTUR/
ANIMAL SCIENCES
uniform diagnostic standards, support training of laboratory scientists/technicians from the member
states, and backstop regional surveillance and epidemiological studies . The activities are also being
supported by the international OIE and FAO reference laboratories like Australian Animal Health
Laboratory, Geelong, Australia . In this regard the National Reference Lab for Poultry Diseases at NARC
has been identified as RLDL for HPAI in this region .
Salient Achievements
"
After completing the required capacity building, the NRLPD is undergoing Accreditation for
ISO-17025 certification through UNIDO. In this regard application has been submitted and
pre- assessment has been completed by PNAC-Islamabad
"
Referral diagnosis was extended to 682 commercial farmers and eight public institutions .
"
22 H9N2 AIVs have been isolated .
"
OIE diagnostic protocols have been standardized at NRLPD and the technical manual has
been prepared accordingly.
"
Flock health monitoring and testing of post-vaccination immune status have been performed
for 948 farms
"
Seroprevalence against AIVs subtypes H5N1, H7N3, H4N6, H3N1 and H9N2 has been
performed using 2660 serum samples received from non vaccinated birds .
"
Organized and conducted an international training for personals of National AI labs in the
SAARC countries (Afghanistan, Bangladesh . Bhutan, India, Maldives, Nepal, Pakistan & Sri
Lanka)
"
Organized two national trainings for harmonization of protocols where 12 persons from all over
the country participated
Future Thrust
"
Continue referral diagnosis services .
Participation in international proficiency testing and provision of proficiency testing to SAARC
countries .
"
Extend trainings to staff from SAARC member states and those from other provinces of
Pakistan .
Acquire international ISO-17025 certification for NRLPD.
Standardization of protocols including PCR, Real Time PCR and sequencing for diagnosis of
new AIV subtype H7N9 .
"
Organize joint vet-med meetings for working in a liaison for joint activity on human-animal
interface .
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 & WILDLIFE
Poultry
Poultry production is considered to be one of the most efficient and economical system of production
of animal protein . In Pakistan Poultry has been playing a vital role in bridging the gap between supply
and demand of animal protein which is an essential food nutrient . Our poultry production is mostly
dependent on exotic commercial lines but ever since the existence of this industry in Pakistan no
attention has been given to our backyard poultry. Indigenous chickens, called desi, have the
advantage of being well adapted to the local environmental conditions (hot cold humid dry and rainy
weather) . These chickens are scavengers by nature and a system of supplementary feeding is not
practiced . 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 economic
importance (like slow growth rate, small egg & body size, poor egg production, late maturity and
broodiness) can be overcome with selective breeding as well as cross breeding with other high
producing exotic breeds . Currently, there are few local breeds like Aseel, Naked Neck and Desi (nondescriptive) which are available for the backyard poultry. This program is thus focusing on
improvement of productive and reproductive performance of backyard poultry in the country.
Research Activities
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 Island
Red, Fayoumi and Aseel breeds . The progeny are being distributed to the villagers for backyard
poultry production . Performance data is shown in Table 5 .
Table 5. Productive performance of various crosses reared at rural poultry farms in and around Islamabad
!No . Characters
Desi
RIR* x
Aseel x WLH
Fayoumi x
WLH**
WLH
1 _ Sexual maturity (days)
21
j
188
224
186
2
Egg count in 52 weeks of production
73
202
143
199
3
Body weight at maturity (grams)
900
1100
1175
1025
~4 (Viability up to 72 weeks of age (%)
90
81
80
83
*RIR, Rhode Island Red **WLH, White Leghorn
Achievements
Around 20,000 highly producing chicks (Figure 8) were supplied to the rural and domestic
poultry farmers. It is expected that as a result of enhanced egg and meat production, the living
standards and health status of the rural population especially the women would increase
significantly 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 .34
million were produced . The sale proceeds were deposited as income .
130
=AKISTAN AGRICULTURAL RESEARCH COUNCII
ANIMAL SCIENCES
Training/Extension Activities
"
After sale services regarding management, vaccination & disease prevention of poultry flocks
through internet . telephone & training at the farm .
More than 50 Research Internee from Veterinary Faculties of AJK, Balochistan, KPK, Punjab
and Sind Universities were trained at Poultry & Wildlife Program.
"
Each month approximately 200 rural/domestic poultry farmers are being entertained for
demand of chicks, management of theirflocks and other entrepreneurial help .
Future Thrust
Expansion of breeding / production & distribution/sale of crossbred chick to the rural poultry
farmers will continue .
"
Mass scale production of these High Producing Rural chicken was started in 2012 . We are
able to increase the production up to 20,000 chicks during the year 2012 . We are planning to
produce approximately 50,000 chicks during the year 2013 .
"
Country level coordinated research project entitled "Genetic Improvement of Rural Poultry
Breeds" was submitted to ALP for funding . The project has been approved by the Technical
Committee and hopefully will be started during the next fiscal year 2013-14
Wildlife
Pakistan has been the land of spectacular scenery, having some of the greatest and beautiful
mountain ranges, rivers, deserts and forests . Due to diversified eco-zone which has had a variety of
habitat 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 . The
major contributors are population explosion, advent of industrial and technological innovations, over
grazing by livestock, land erosion by wind and water, deforestation and indiscriminate hunting and
poaching etc .
In the mid-1800s, elephant, lion, and tiger were common in present Pakistan . Herds of Chinkara were
common 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 Kirthar
Range, slopes of Northern Areas mountains and many areas of Balochistan . Ibex and Markhor were
found in most of the ranges of northern mountains and Balochistan and hog deer in all riverine tracts of
Sindh and Punjab . Nilgai was commonly found in agricultural lands from Bahawalpur to Sialkot and so
was Kashmir stag in the Neelam Valley of Azad Jammu and Kashmir. Musk deer inhabited almost all
birch 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 the
nation's other natural and agricultural ecosystems . Hence, there is need to suggest necessary steps
for its conservation/management . Key to protect biological heritage of Pakistan is in the involvement of
local people and in the support from research institutions, like PARC for conservation and sustainable
use of biodiversity. Lack of scientific data about the biology and ecology of these species from their
natural habitat and captivity is a major constraint in their scientific management . In order to conserve
and manage this natural resource, in nature and in captivity, data on biology ecology and behavior of
wildlife species is needed but it is very difficult to record the exact observations of some wildlife
species due to their peculiar behavior /shy habits in their inaccessible natural habitat and it remains the
main 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 of
indigenous endangered wild animals and birds
Keeping in view above referred problems and hurdles in management and conservation of native
wildlife species a program was proposed for breeding of wild animals and birds in captivity. Data about
their breeding, behavior and feeding is being collected . Feed preference experiment has been
initiated . Seasonal fodder cropping continued for animal feeding and planted fruit trees and shade
plants inside the section. We have achieved successful breeding of Hog deer (Axis porcinus) which is
vulnerable according to IUCN Red list 2008 . Data on breeding behavior and growth characteristics etc
is projected briefly in Table 6 .
Table 6. Breeding behavior and growth characteristics of Hog Deer in captivity at NARC
Sexual Activity in
Captivity : l s ` sign
ticxual & plating in
Captivity
Male : 09-10 months
Female : 10-11 months
Sexual hchti~ for ~ca, obser~ cd froth Aril to October.
Male buck was more active during the months of April & May .
Most mating was observed after the onset of spring and before extreme
humid hot climate .
Copulation observed in morning and this observation was recorded twice at
our vicinity.
Gestation Period
Black Buck, 5 Months
Hog Deer : 8 Months
Fawning Season
Black Buck : March, September . llog Deer : February-March, November-Dec .
Reproductive
No reproductive disorder observed in hog deer female
Disorders
_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"ear3
Antlers %weight (year-t\ ise)
?00 gram bout
1
800 gram both
850 gram both
Antler-1
Antler-2
Antler-1
Antler-2
Antler-1
Antler-2
Length (cm)
40cm
39cm
39ctn
38cm
42cm
43cm
Antler Cycle
Period (peak) of casting
6 months
Span of antler growth
6 months
Velvet stripping
July-Aug
Achievements
Construction of 5 deer enclosures, 8 bird cages. one bird aviary, office/store building is
completed
Developed permanent pasture fields and plantation inside wildlife research section for feeding
needs and shades.
Successful breeding results of Hog deer and Peafowl achieved
Future Thrust
"
Maintain already established wildlife conservation/breeding research facilities at NARC
(Figure 9-11) .
"
Transfer/sharing of technology to researchers, students, wildlife farmers, breeders and wildlife
management and conservation related organizations & NGOs .
13 2
PAKISTAN AGRICULTURAL RESEARCH COUNCIL
ANIMAL SCIENCES
"
Collaboration for joint research with Wildlife Management Department UAAR, Zoology
Department Punjab University, Wildlife Department, University of Veterinary and Animal
Sciences, Lahore, WWF Pakistan, Punjab Wildlife Department, Environmental Department
CDA 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
Figure 13: Black Buck Deer (Male) at Wildlife Section
Figure 14 : Hog Deer (Male) at Wildlife Section
ANIMAL BIOTECHNOLOGY
Productive traits of milk animals, both qualitative and quantitative, are primarily determined by their
genetic make up . In this context several genes have been identified and this information, obtained
from DNA loci affecting economic traits, is helping in better selection of livestock breeds . In the milk
progeny testing programs of bulls we have to wait for seven to eight years before we can select the
bulls on the basis of performance of their female progeny. If we can develop some DNA test that can
help us to identify animals having good productive genes as soon as it is born then it will save both
time and rearing cost of productive farm animals.
Dairy cattle breeding programs in Pakistan involve utilization of imported/exotic semen from elite bulls
with high genetic merit for the dissemination of alleles associated with positive economic traits, such
as increased milk yield . Introduction and dissemination of such traits is beneficial for animal
production but likelihood of simultaneous incorporation of defective genes is increased which is
undesirable . Under present trend of short term livestock improvement culture, Holstein-Friesian breed
is widely disseminated throughout the country, with regional populations genetically linked because of
exotic semen, embryos and live animals imported from abroad . Therefore, defective alleles occurring
within widely-used breeding lines are likely to be present in the Pakistan Holstein-Friesian and
crossbred population thereto.
Therefore, current research activities at NIGAB are focusing on identification of animals that have
greater production potential and are free from genetic anomalies . Similarly, molecular
characterization of pathogens of food animal origin and identification and development of
immunogenic proteins required for diagnostics and vaccine production is also actively persuaded .
Research Activities
a.
Genetic Improvement of Production Traits in Indigenous Livestock Breeds
Experiment :
Molecular analysis of genes affecting milk production traits and disease
resistance in food animals.
Bovine and ovine genomes are being analyzed internationally for economic traits . Modern cattle
breeding strategies involve identification of milk, meat & disease resistance genes and its variants
that can be incorporated into selection programs . In this regard several genes have been
identified . More than 26 genes have been mapped and characterized for milk production traits and
most important include DGAT1, CSN1 S1, CSN1 S3, PRL, LEP STAT5A, BoLA-DRB3 etc. The
objective of NIGAB is to undertake research studies focusing on identification of genotypes with
greater potential production using molecular genetic approaches .
Salient Achievements
"
Gene polymorphism for DGAT I, rc-CN and PRL genes related to milk fat and protein in NiliRavi buffalo, Sahiwal and Achai cattle has been analyzed . The results indicated that the
animals of Nili-Ravi buffaloes are homozygous for DGAT I (K allele), K-CN (B-allele) and
PRL (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.
ANIMAL SCIENCES
"
Analyzed 500 Nili-Ravi buffalo samples collected for FMDV detection and their persistence
using Real Time PCR.
b. Genetic Status of Pakistan Cattle Population for Inherited Disorders
Experiment :
Genetic testing for inherited disorders in Pakistani Bos indicus, Bos taurus
and their cross breed.
Inherited disorders are of major importance in Holstein Frisian cattle a breed that now dominates
the world dairy industry due to high milk production . The important inherited disorder BLAD was
disseminated globally in 1992 by this breed and reported in Pakistani Holstein Friesians in 2008
and 2012 . However a range of inherited disorders recognized internationally are still in need to
explore in this breed in Pakistan . Therefore, it was deemed important to initiate inherited disease
detection by DNA testing in Holstein Friesian and its crossbred population in Pakistan to
strengthen the National breed improvement programs . (Figure-14)
Salient Achievements
c.
"
PCR Optimization for the amplification of gene encoding bovine CD18 and PCR amplification
of the target genes responsible for the Citrullinaemia is achieved . Similarly amplification of
selected regions of UMPS and SLC39A4 genes containing 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 is
achieved .
"
Screening for the citrullinaemia and BLAD is in process . Screened 200 samples including AI
bulls and seen some mutations trying to confirm through sequencing .
Development of DNA Based Diagnostics
Experiment :
Standardization of DNA based diagnostic test for rapid detection of foot and
mouth disease virus (FMDV) under laboratory and field conditions .
Foot and mouth disease (FMD) usually results in severe economic losses and impact on both
national and international trade within the livestock and animal products . Conventional molecular
techniques for the detection of FMDV are expensive and time consuming so a novel nucleic acid
amplification method, termed reverse transcription loop-mediated isothermal amplification (RT
LAMP) is being developed for FMD diagnosis and serotype differentiation .
Salient Achievements
"
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 cell
lines .
"
Standardized Lamp PCR for FMD virus detection.
"
For FMD persistence studies . 100 samples were tested out of these 11 found positive . Results
showed apparently healthy animals carry FMDV
d.
Recombinant Protein for Therapeutics and Vaccine Development
Experiment :
Development of cloned vaccines against avian adenovirus serotype-4 in
poultry.
Fowl adenovirus infections are widespread inmost of the poultry raising countries of the world. Fowl
adenoviruses belonging to group I have been associated with a number of clinical conditions like
effect on egg production and Hydropericardium syndrome which is an emerging disease of broilers
and breeders resulting in heavy economic losses . Autogeneous formalin inactivated vaccines
prepared from infected liver homogenate have been used against HPS but they are unable to
control the disease So present study was designed to develop a cloned vaccine against HPS
Salient Achievements
"
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 and
confirmed by PCR and sequencing .
"
Recombinant protein was expressed in BL21 cells and purified by GST tagged columns using
GST buffer kit.
Training/Extension/Other Activities
Research Publications
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 its
association 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 advanced
technologies" organized by NIBGE, Faisalabad on 31 "January -I" February, 2013 .
"
Attended a three-week training on "Real Time PCR based detection of FMDW organized by
ARD-USDA at AHP ASI, NARC in December, 2012 .
"
Participated in SAARC Regional Conference on New Frontiers in Agricultural Genomics and
Biotechnology, 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 cell
culture 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 with
commercial vaccine . Asma Jabeen, Khalid Naeem, Tahira Kamal and S .M . Saqlan Naqvi . 6`h
Annual Cell Culture Technology event Recent Advances, Future Prospects March 2013 Royal
College of Pathologists, London .
"
Tahira Kamal, K. Khalid Naeem. Amir bin Zahur, Ghulam Muhammad Ali, Naeem Riaz, Fozia
Nasreen, Aman Ullah and Asma Latif . (2013) . Comparison for the sensitivity of BHK-21 and
Bovine Kidney Cell Line for the Isolation of FMD viruses . Presented at 9th International Biennial
Conference of Pakistan Society for Micvrobiology, January 28-31, 2013 . Karachi-Pakistan .
"
Evidence of foot and mouth disease virus in apparently healthy buffaloes in abattoirs of
Islamabad 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 Pakistan
Society 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) . K232A
polymorphism in Diacylglycerol Acyl transferase I gene in Nili-Ravi buffalo, Sahiwal & Achai
Cattle . SAARC Regional Conference on 'New Frontiers in Agricultural Genomics and
Biotechnology 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 Bos
indicus, 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 in
Agricultural Genomics and Biotechnology" hosted by NIGAB at Marriott Hotel Islamabad from
June 5-7. 2012 .
Future Thrust
Standardization of PCR forthe amplification of defensin gene and to study DNA polymorphism
related 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 to
study association with the milk production traits .
"
Screening work, for the diagnosis of inherited disorders (BLAD, Citrullinaemia, DUMPS and
BHZD) from Punjab area will be performed during the year 2013-2014 .
"
A proposal is submitted for research funding as Co-PI entitled "Identification and
Authentication of Market Meat and Meat Products by Biological & Molecular approaches" to
RADP 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 .
M1,1 .a4e0-svInu11u14IsIs11 .1
-
x»411 .~ gar
" ~Ir
"~rlw W " ri+rri+r~ss . .
Prolactin gene polymorphism in
Buffalo & Cattle breeds
Chicken prolactin gene polymorphism
Figure 15 : Phylogenetic tree analysis (Fasciola spp) of FG ITS-2 constructed using Bayesian methods indicates
that FG ITS2 found in Pakistan is unique from the FG ITS2 found at other places .
BLAD
Amplification by Sl primers
ITGB2 gene by Rl primers
M
M
1
2
3
4
5 -Ve
M
111 ; I Illlbp ladder
~i
58bp
M
C.383A>G, p.D128G (Heterozygous)
c
c
=
1
..
_
R
c
c
i
=
r
d-
iiiiiiiiiiio
-1-
r" "- -
-1+
a+
We
¢° 619bp
-1-
-I-
-
G
DNA sequence chromatogram
Taq I endonuclease restriction analysis
Figure 16: Genetic analysis of Bovine leukocyte adhesion deficiency in HF and its crossbred
138
ANIMAL SCIENCES
AQUACULTURE & FISHERIES
Aquaculture is currently one of the fastest growing food production systems in the world with
production level increasing at an average rate of 11 percent per year over the last decade . In Pakistan
too, the major potential for increase in fish production lies in inland aquaculture . Therefore, there is an
increased interest in the development of aquaculture production not only for the food security but also
for 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 of
necessary nutrition and effective disease control measures . The main objective of fisheries research is
to promote aquaculture and fisheries productivity by increasing per unit area production through
intensive 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 as
development of ornamental fish science and technology that has substantial local and overseas
markets .
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 the
possibilities of its farming within the country. For this purpose, a study has been carried out on some
fresh 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 as
well as culturists . The success of ornamental fish culture in the country will not only meet the local
demand but will also provide the opportunity to raise foreign exchange for country through it export in
the regions where there is high demand . In fish culture diet is major economic factor determining
profitability. 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 (orange
and white) (Cyprinus carpio) and Hi fin Koi Carp (C . carpio) from localy bred stock has been stocked in
eathern 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 a
replacement 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 (29
pairs) has been stocked in earthen ponds, raceways and circular fiber glass tanks for breeding trials
during November 2012.
Breeding season of ornamental fishes has been determine such Double tail (Carassius auratus
auratus) breeds in December, Shubinkin (C . auratus) breeds in January, Local Koi Carp (orange and
white) (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 (orange
and white) (Cyprinus carpio), Hi fin Koi Carp (C . carpio), Mollies and Guppies has been achieved
successfully.
Experiment 3 :
Induced breeding of ornamental fish .
Induced breeding ensures the availability of seed during the whole breeding season. In this regards
successful 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 color enhancer.
Three different diets supplemented with 1) spirulina (blue green alga), 2) red beet root and 3) carrot
were compared with locally developed diet (30 % CP) for color enhancement and augmented
aesthetic appeal.
Diets supplemented with red beet root were found more effective as color enhancer whereas mixture
of above three diets showed further better result .
Training/Extension Activities
Exhibitions of ornamental fish technology at FAO Kisan Mela at NARC, FAO World food Day at NARC
and Autumn and Spring Festival at Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi for public
awareness 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 been
transplanted to different farmers, Directorate of Fisheries Khyber Pakhtunkhwa, Peshawar, Directorate
of Fisheries, Gilgit-Baltistan and Punjab Fisheries Department, Lahore .
Culture and breeding technology of ornamental fish (Shubunkin and High fin Koi carp) has been
transferred to 5 progressive farmers of Potohar Region (Jand, district Attock) and successful breeding
has 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 farm
ponds using different management inputs
This project has been designed to increase the fish production through intensification of fish
culture and timely inputs utilization such as fertilizer and artificial diets . Judicious use of such
ANIMAL SCIENCES
inputs and adoption of the intensive fish culture technology can easily enhance the fish production
from present level of about 1000-1200 kg/acre to about 1800-2500 kg/acre per year as it has been
already 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 by
stocking participating farms in all three provinces with three indigenous or major carp, namely
rohu (Labeo rohita), mrigal (Cirrhinus mrigala) and Catla (Catla catla) and three exotic Chinese
carps i .e., grass carp (Ctenopharyngodon idella), silver carp (Hypophthalmichthys molitrix) and
common carp (Cyprinus carpio) .
Figure 17: Brooders of Shubinkin
(Carassius auratus)
Figure 18 : Brooder of Double tail
(Carassius auratus)
Figure 19 : Brooders of Local koi Carp
(Cyprinus carpio)
f
Figure 20: Brooders of Hi fin koi carp Figure 21 : Shubunkin
(Cyprinus carpio)
(Carassius auratus Fry
Figure 23 : High fin Koi carp
(Cyprinus carpio) fry
Figure 22: Double Tail
(Oranda) (Carassiusa auratus) Fry
Figure 24 : Koi Carp white
(Cyprinus carpio) Fry
Salient Achievement
The project has been initiated recently w.e.f. 01-10-2012 in four province of country and in
NARC . The coordination has been developed between four provinces and PARC .
"
Survey was conducted to identify the progressive fish farmers' at Ali Pur Chattha, Gujranwala
district for the selection of fish farms . A farm was selected at Jamkay Chattha, Gujranwala for
intensive fish culture.
"
The fish were stocked at farmers' field at different stocking density i .e . 1500 fish / acre . 1800
fish/ acre and 2100 fish/acre and was supplement with 20% CP artificial diet . The average
weight gain aftertwo months is 120 gm .
Training/Extension Activities
"
Artificial diet developed for all developmental stages of carps at Aquaculture and Fisheries
Program and transferred to farmers in Intensive culture project
Future Thrust
"
Study growth of carps fed on different supplementary fish feeds and testing of diets indifferent
agro-ecological zones .
Evaluate varying stocking densities of carps.