Académique Documents
Professionnel Documents
Culture Documents
Government of lndia
Ministry of Agriculture and Farmers Welfare
Department of Animal Husbandry & Dairying
a$*s\:'fa\1
(Bhushan Tyagi)
Assistant Commissioner (AH)
Distribution to:
(D Principal Secretary/Secretary , Department of Animal Husbandry, All state
Governments
(ii) Executive Director, NDDB, Anand
(iii) Managing Directors, AII State Milk Federations
(iv) Director , Department of Animal Husbandry,
All state Governments
(v) CEO/MD, State Livestock Development Board
(vi) ln-Charge of all Semen Stations
(vli) Chairman, CMU
(viii) Member Secretary, CMU
GOVERNMENT OF INDIA
MINISTRY OF AGRICUTTURE AND FARMERS WELFARE
DEPARTMENT OF ANIMAL HUSBANDRY & DAIRYING
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osp ileLls Jovo 'fulunoc eql ut suouels uewes rc! locoloJd prepuels
wnwtutw pue locoloJd uoque^eld asees/o onss/ //eq{s lovo eql,,
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og pa1sa66ns arern 6uuvro;;o; pue aut;aptn6paplo Uetp aq1 o1 paat6e asnoq aq1
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pue n!!3aql ,{q saseasrp lle roJ aalJ aseaslp pauluac ale {aq1 lll} l3e aql Jepun
'ureql lsure6e uorlce alel lleqs retruJo 1ue1adu.toc7.to1corlo aql pue '6002 ]cv
voclcd Jo 0z uortcas rapun ,seaJe palceJul, se peleall aq ol aJe suouels uaulas
paJolsr6arun llv 'sasop uaues ales / Ilddns ol aq lleqs suollels uaulos
pa/\Aolle
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constitute centrar Monitoing unit for evaruation and
monitoring of semen
sfaf/ons which shall among others, consisls of expefts
and at least one
representative of concerned State Government not
below the rank of
D i re cto r/J o i nt D i rect o r/E q u i v a I e n t. "
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Chairman, CMU for evaluation of Semen Station, explained the existing Minimum
Standards Protocol (MSP) and the proposed revision in the MSP for Semen Production.
After detailed deliberation the modification in the MSP was accepted by the house
which is placed at Annexur*ll. The major revisions incorporated are as follows:
. Rework on the table made for assessing the Bulls genetic worth in revised MSP
and send for consent to all stakeholder.
o Karyotyping should continue as it is once in life time test.
. Minimum period between tlvo ejaculate should be at least 15 min.
. lt is recommended to preferably collect two ejaculate per day collection and two
days of collection per week from a healthy bull.
o Pattern of Printing Semen Straw from left to right should be clearly stated in the
MSP document.
. Quality Control Test to be conducted daily.
. lt is suggested to modify the manpower requirement as "lt is recommended that
Semen station in-charge should have at-least 5 years experiences in bovine
semen production and should not be transferred frequently. lncumbent should be
positioned for 5 years."
. All Semen Stations not to conduct MSP disease testing of their Bull Herd from
their own state lab or their own agency. The MSP disease testing should be done
by third party. CMU to only consider results of third party disease testing of bulls.
. Request has come to include IBR testing of Semen doses through RT-PCR by
semen station itself. DADF will take final call on it.
o ICAR-|VR| not regularly supplying antigen for testing TB and JD. DADF to take
up the issue with ICAR.
Director, CFSP& Tl, Hessarghata, Karnataka presented the report of CMU evaluation
and the evaluation status of the Al Training lnstitutes.
l4 l-
Assistant commissioner, Nationar Livestock Mission, DADF
exprained the Minimum
standards Protocor for semen production in smal Ruminants
and standard operating
Procedure for Ar in Goats. Folowing recommendations
were made by participants of
the workshop :
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ATTENDANC ES HEET
WORKSHOP ON SEMEN PRODUCT]ON
16111 JANUARY 201g
NASC COMPLEX, PUSA, NEW DELHI
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16IH JANUARY 2019
NASC COMPLEX PUSA NEW DELHI
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ATTENDANCE SHEET
WORKSHOP ON SEMEN PRODUCTION
16" JANUARY 20t9
NASC COMPLEX PUSA, NEW DELHI
'besignati6n
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Minimum Standards
For
Production of Bovine Frozen Semen
2019
Page I of 75
h'/ a IIMU FOR PRODUCTIO N OF BOVINE FROZEN SEMEN
Artificial Insemination with frozen semen has been proved to be the
best toor
worldwide for mass genetic improvement through dissemination
of superior
germplasm. This objective can be achieved only if the frozen
semen used in
A'I programme conforms to certain prescribed quality standards.
For
production of quality semen, it is most important that
the bulls used in A.I
programme meet prescribed norms, and are disease-
free and that the
semen is harvested and processed in accordance with the
standard
protocols' The minimum essentiar protocols required
for production of
qua-lity semen are described in t].is manual. Fa ure
to observe these
guidelines may result in production of poor quality semen
making it unsafe
as well as unfit for use in breeding through artificial insemination
programme.
Page 2 of75
As a very few bulls coming from genetic improvement programmes are being
inducted in the semen stations at present, it would not be possible to have
breeding values for all bulls of different breeds maintained by the semen
stations. However to ensure that high quality genetics get into the semen
stations, a simplilied procedure giving weightage to different ways the bulls
are obtained could be adopted to assess the genetic worth of a semen
station. At present bulls are obtained in the following different ways:
Weightages proposed for the above mentioned six different ways of obtaining
young bulls and a procedure for calculating a genetic worth (out of 100
points) of a semen station having bulls obtained from the six sources is
shown in the Table below:
Page 3 of 75
Table: 1 Weightage for different categories of bu[ for calculating
score
for genetics in CMU evaluation
No. of
bulls at 7o of bulls
s the of I
semen dillerent \lleighted
N Source of bulls station categories trIeightage I
record gg
5 Mother single
record and sire 50 16.7 0.0
known 0.0
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6 Mother single record and sire
not known
10
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3.3 0.o I o.o I
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Total bulls 300 I
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loo.o I
56.71
Genomic Breeding Value to be includ ed a-fter complete
development
and va-lidation of Genomic Serection chip by a committee
constituted
by DADF.
The score card prepared for cMU Evaruation shalr have
at least 5o%
weightage of total score for the Genetic worth of the
Semen station.
Page 4 ol75
Note: " PT projects follow the GoI'PT SOP; top 10%o of active proven bulls
and top 57o of recorded females based on breeding values (provided by
Breeding Value Estimation Committee constituted by Govt. of India.) are
used for selection of young bulls
"Bull's sire is an HGM bull and mother is from top 1O% of recorded females
(Minimum 50O females under recording either from institutional farms or
fie1d)"
** PS projects follow GoI's PS SOP; top 5% recorded females and HGM bulls
are used for producing young bull calves or bulls obtained from bull mother
farms.
$ BuU's sire is an HGM bull and mother is from top 10% of recorded females
(Minimum 50O females under recording)
$$ lvtother is from top 10% recorded females (Minimum 5OO females under
recording)
Page 5 of 75
- If a bull is produced through ET, the parameters applicable
for sire and
dam should be used for putting the buls in different categories i.e.
if the
sire is among top proven bulls and donor is selected based on top
recorded
dams, the bull should be put in category 1 and so on.
(This arrangement ui come in force u).e.f. lst Apit 2020, after compretion
of
remaining 2017-18 eualuation uhich u,till be based on unreuised
MSp (2012)).
Page 6 of 75
Amritmahal 400 500 4.O
The sta-ndard for Dam's lactation yield for Fl cross bulls will be the same as
that of respective indigenous bull dam i.e. Gir, Sahiwal, Kankrej, Red
Sindhi, etc.
For Breeds not mentioned in above table, concerned state government may
notify the min. Dam's lactation details and Breed code.
For imported bulls and embryos, tle standards for import of qerrnplasm as
prescribed in the "Guidelines for export / imoort of bouine dermDlasm" issued
and amended from time -to -time bu DADF, Ministru of Aqianlfure and
Fanners Welfare, GOI shall be applicable.
Page 7 of 75
compatible for registration under INAPH, presently being managed by
NDDB, Anand.
b). The bar coded ear tag with unique 12 digit raser printed number
w l be
physica-lly applied to the bun and will remain on it for rife.
If the burl is sold
to other SS, tJle tag no. i.e the bull identification will remain
unchanged.
c)' In case the tag falrs, is 10st or destroyed, it will be changed
with a new 12
digit Bull ID following standard procedure of "Ear Tag change, under
INAPH.
Page 8 of 75
measurement of scrotal circumference and body weight needs to be initiated
immediately.
4. Quarantine
A minimum quarantine period of 60 days is compulsory before bringing new
bulls into a semen station. Only after favourable results from the health
control point, the bulls shall be admitted to the semen station. Relevant
definitions are given in Annexure- I
5. Testing of Bulls
The testing protocols/ procedures for bulls against T\rberculosis,
Johne,s
disease, Brucellosis, campylobacteriosis, Trichomoniasis,
Infectious Bovine
Rhinotracheitis and Bovine Viral Diarrhoea are given in
Annexure- 4 to ro.
The breeding burls shourd be free from above mentioned
diseases. Though
Johne's disease is not a sexually transmitted disease
but being a chronic,
infectious and incurable disease, it has been included
and the breeding
bulls found positive for Johne's need to be removed. The
bul1s in the
quaraatine/ rearing station and the resident herd
should go through
periodical testing and vaccinations as per the schedule
listed in the manual.
6. Vaccination Schedule
Page 10 of 75
Guidelines issued bv the De T) artment of Animai Husband T_\/ and D a 1n:t n g.
Ministrv of Aericulture and F armer Welfare for progres sive IBR BVD control
and as amended from time-to-time shall be follow edi n letter and spirit bv all
semen stations.
Besides, the semen station are advised to cull those bulls which have
completed eight years of productive period or 3 lakh semen doses, whichever
is earlier unless the bull is of exceptional genetic merit. In addition, the
bulls with poor libido, poor semen quality, incurable lameness, etc. may also
be culled on regular basis.
8. Housing
Bull sheds shall have spacious individual pens with adequate loafing area,
manger and water trough with access to drinking water all time. Adequate
shade around the bull shed shall be provided. The roof shall be made of
asbestos or suitable materials. During summer, cooling system with
sprinklers and fans is required particularly for the buffaloes and exotic
bulls. Disinfectants like formalin or phenyl based compounds shall trot be
used in the bull sheds. Alternatively, compounds containing Gluteraldehyde
may be used. Weekly spraying of Sodium Carbonate (4% solution) shall also
be practiced. The floor should be sterilized at least once a year by a blow
lamp or by burning straws. At one corner of the farm, there shall be an
isolation shed for separating ailing / sick bull(s) for treatment. Bull(s) once
diagnosed suffering from any of the infectious diseases shall be removed
immediately from semen station to contain its spread to other bulls.
9. Maoagement of Bulls
The semen station shall arrange for carr5ring out ring vaccinations of all
cloven footed animals including swine against FMD within a radius of 10 km
around the semen station. Vaccinations against HS and BQ shall be carried
out in the areas having incidence ofthese diseases.
I
scientihc method
I
Page 12 of75
mounting area shall have sand and limestone mixture for proper
footing of bulls. Alternatively, good quality rubber mat (with
interlocking arrangement) or coir mat shall be put into concrete
groove of the mounting area for adequate cushioning effect. After
collection, the area must be thoroughly cleaned and odorless
disinfectant solution (Colloidal iodine) be sprayed. A dusty floor shall
be avoided to prevent dust falling on the A.V / semen samples.
c) The person carrying out preputial wash must use disposable gloves
and separate sterilized oozzle for each bull to avoid transmission of
infection from one bu11 to another. Preputial washing should be
carried out if the bull is soiled otherwise it is better to avoid.
Page l5 of 75
g) Sterilized bull aprons shall be used to avoid penis touching
hindquarter of the dummy.
h) Before every collection, the semen collector shall wash his hands with
suitable antiseptic solution or use disposable gloves or do both. The
semen collector shall not touch the penis.
i) Semen should be collected either by the Veterinarian himself or by a
suitably trained technician / staff under his close supervision. while
taking collection, it shall be ensured that AV is not thrust on penis of
bull, instead the penis is gently guided into the AV.
k) Appropriate size AVs, ranging from g-14",shal be used for cattle and
buffaloes ensuring that the semen is ejaculated in the cone. The cone
shall be of good quality Neoprene rubber/Silicone rubber.
Page 16 of 75
m) The AV shall not to be shaken after ejaculation and carried to the pass
box with open end slightly inclined downward to avoid flowing down
and mixing of iubricant and debris with the semen samples.
o) The entry of visitors and staff /labourers (otJler than those involved in
semen collection) shall be strictly prohibited in the collection arena at
the time of semen collection.
Page 17 of 75
b) The ceiling and walls of the laboratory shall be made up of non_
porous materials. A11 cracks and crevices shall be sealed to control
pests ald insects.
c) Entry of persons to the laboratory, other than laboratory
personnel, shall be strictly restricted. Airlock system or anti_room
shall be provided to avoid direct entry to the semen_processing
laboratory.
e) Preferably cassette tlrpe or, split type air conditioners fitted with air
puriffing system with remote temperature control mechanism
should be installed to maintain the room temperature at 2O"C _
Page 18 of75
0 Sink drains shall be decontaminated routinely with a disinfectant.
Sink shall not be placed in the semen processing room.
Page 19 of 75
l) The working platforms, the exposed parts of equipments and other
items to be handled during processing of semen, shall be cleaned
with alcohol (Iso Propvll or Glutaril. It is advisable to repeat
7 Oo/o
11 (Bl Equipment
Page 20 of 75
b) The pre-filter of Laminar Airflow unit shall be cleaned Ouarterlv.
Routine servicing and DOP testing twice a year will ensure efliciency
of HEPA filters. Alternatively, culture plate test to monitor bacterial
Ioad of the air passed through the filters shall be carried out at weekly
interva-ls to assess its efficacy/ functioning.
c) Digital photometer / Computer aided Spectrophotometer shall be
validated with Haemoc5.tometer readings for sperm concentration
twice a year separately for cattle and buffalo (20 samples each).
d) The automatic semen straw hlling and sealing machine sha-ll be
thoroughly cleaned, immediately after use.
Page 2l of 75
i) The Liquid Nitrogen containers returned / received from outside shall
be disinfected thoroughly with 4%o sodium carbonate solution and
frnally with I to 4o/o formaldehyde.
k) The refrigerator meant for storing eggs, a.tibiotics and buffer shall
only for storing t-hese materials. while vaccines, medicines and other
materials shall be stored at a place away from the semen laboratory.
The refrigerator used for storing eggs, etc. shall be cleaned and
sterilized every week using alcohol swab.
Page 22 of 75
11 (Cf Personnel Hygiere
Clothing, skin and hair of laboratory personnel are the recognized
sources of contamination. Hence, everyone should wear laboratory
aprons, footwear and take other necessary precautions all the time while
working in the laboratory. Hands shall be washed with soap and water
and rinsed witn' 7 Oo/" alcohol before commencing work in the laboratory.
The other staff working in farm should be tested for TB and Brucellosis
once in two years.
11 (Df Diluents
d) Glassware, collection tubes, etc. shall not be held from their rim /
mouth.
Page 23 of75
f) After adding all the components of buffer viz. TRIS, Citric Acid,
Glycerol and Fructose in ultra-pure water (Autoclaved) or triple glass/
double distilled water it should be sterilized by microhltration
(Optional). If the buffer is prepared on the previous day, it should be
stored in the refrigerator and antibiotics are to be added next dav in
the morning after warming it to 34.C.
h) Only fresh eggs shall be used for making dilutor. The eggs shall be
stored in refrigerator after wiping with dry cotton. Just before preparation
of dilutor, eggs shall be wiped with ZO%o alcohol (Iso propyl). To ensure
the quality, eggs shall be purchased from known sources.
Page 24 of 75
b) As soon as the neat semen is received, it shall be kept in a thermo-
controlled water bath at 34o C under Laminar Air Flow Unit, after
recording the volume of semen.
After the linal dilution the flask should be kept at room temperature
for lO-15 minutes so that temperature reaches 2O-22"C.
Page 25 of75
e) Semen samples selected for processing should have a minimum of
7 Oo/o initial progressive motility.
f) Filling and sealing of semen shall be done under Laminar Air Flow
Unit using sterile straws, frlling nozzles and disposable rubber
tubing's. Rubber tubing's sha-ll not be reused in any case.
g) unused straws shall be repacked (air-tight) under Laminar Air Flow
Unit before storage. Immediately after use, all the glass ware and
other re-usable articles shall be immersed in lukewarm neu trai
detergent solution (in a plastic tub near the Laminar Air Flow Unit).
All semen stations shall folrow the foilowing colour codes for filiing of
semen in straws:
Table: 3 Colour Specifications
Breed Colour
Holstein Pink/Rose
I
Jersey Yellow
I
I
Page 26 of 75
Sunandini Blue
Buffalo Grey
Each Semen Station shall print certain key information on the straw in the
following Sequence, starting from the factory end.
1. SSBull ID - Alphanumeric
2. Breed of Bull
3. Semen Station name
4. Batch No with Ejaculation No in Parentheses
5. Dam's Lactation Yield and source ( e.g. 3380- PT)
6. Brand image
7. Brand Code
2 AMUL I
Gujarat Coop. AMU
I
Page27 of75
3 BAIF Maharashtra Trust BAF
Page 28 of75
24 Aurangabad I Maharashtra Govt AUR
30 Cuttack
I
I oaisna To.*. CTK
I
a1
Deep Frozen Semen Uttar Pradesh Govt. RKH
Station,Rehmankhera,
Lucknow
34 Gurgaon Haryana I
Govt GUR
I
I
I
J/ Jagadhari I
Haryana Govt. JAG
Page29 of75
+J Pune Maharashtra Govt. PNE
I
I
Kashmir
48 SSCC, Hessarghatta Karnataka Govt. SSC
I
I
Ahmednagar.
I
Page 30 of 75
Sr.No. SS Bull ID Breed Code formation
2 If there is no cross with the breed then that breed has lo0o/o blood
Ievel of its own. For e.g Breed code for 1007o pure Jersey animal will
be 'JY' . Therefore, SS Bull ID of a 1007o pure jersey bull at ABC
Semen Station will be ABC-JY-34'.
1
For crossbred, having two or more than two type of breed involve,
Breed code and SS BuI1 ID will be formed in the following order.
(b) Incase, exotic breed is not available then breed code of the
indigenous breed having maximum Toage will be considered and
concatenated with 'CB'. For e.g. a crossbred bull having
Sahiwal(SO%), Red Sindhi(2S%) and Gir(2\o/ol breed will have breed
code 'SHCB'and SS Bull ID as ABC-SHCB-97'.
Buffalo
1 Bhadawari BW
2 Buffalo Jaffarabadi JF
\) Buffalo Marathwadi MD
4 Buffaio Mehsana MH
Page 3l of 75
s.N. SpeeiesName BreedName CODE
5 Buffalo Murrah MR
6 Buffalo Nagpuri NP
7 Buffalo Nili-Ravi NR
8 Buffalo Pandharpuri PD
9 Buffalo Surti SU
10 Buffalo Toda TD
11 Buffalo Banni BN
12 Buffalo Chilika CK
13 Buffalo Kalahandi KD
).4 Buffalo Bargur BG
15 Buffalo Luit(Swam p) LU
16 Cattle HF HF
17 Cattle Jersey JY
18 Cattle Amritmahal AM
19 Cattle Bachaur BC
20 Cattle Bargur BR
2t Cattle Dangi DN
22 Cattle Deoni DO
23 Cattle Gaolao GL
24 Cattle Gir GR
25 Cattle Hallikar HK
26 Cattie Hariana HR
27 Cattle Kangayam KY
28 Cattle Kankrej KN
29 Cattle Kenkantha KK
30 Cattle Kherigarh KG
31 Cattle Khillar KH
32 Cattle Krishna Valley KV
JJ Cattle Malvi MV
34 Cattle Mewati MW
35 Cattle Nagori NG
36 Cattle Nimari NM
Cattle On le OG
38 Cattle Ponwar PW
39 Cattle Punganur PG
40 Cattle Rathi RT
Page 32 of 75
41 Cattle Red Kandhari RK
42 Cattle Red Sindhi RS
43 Cattle Sahiwal SH
44 Cattle Siri SR
45 Cattle Tharparkar TH
46 Cattle Umblachery UB
A7 Cattle Vechur VC
48 Cattle Motu MO
49 Cattle Ghumusari GH
50 Cattle Binjharpuri BH
51 Cattle Khariar KR
52 Cattle Pulikulam PU
53 Cattle Kosali KS
54 Cattle Malnad Gidda MG
55 Cattle Belahi BL
56 Cattle Gangatiri GT
57 Cattle Badri BD
58 Cattle Ladakhi LD
59 Cattle Konkan Kapila KP
60 Cattle Lakhimi LK
Page 33 of 75
discarded. semen samples with circular a,d Jerky movements and with
any other abnormal motitity shall be discarded.
The quality control measures are sub divided into mandatory and
optional tests and include continuous monitoring of semen production at
various steps/sub processes essential for quality semen production. It shall
be mandatory to document quarterly suamary of each test including the
number of samples tested and the number of sampres not meeting
standard with the follow up action taken, The quarterly summary will
be authenticated by the Officer iu-charge ofthe Semen Station.
Mandatory Testing
no. test to be
I carried out I
Page 34 of 75
1.1(af Sterilization Microbial Daily, Failed test -Re-
[,oad random wash and steril2e
I
Testing of sampling, a the whole lot of
Glassware minimum of Glassware again on
two glass the date of result
wares assuming that the
same error in
sterilization process
I I is continuing.
I
Testing sampling of a I
the whole lot of AVs
I
I of AV Wash minimum of again on the date of
I I two ready to result assuming that
I
I
use AVs the same error in
I
sterilization process
is continuing.
Page 35 of 75
1.2(af Calibration of Annual Outsourced
I
to be taken up with
I
I I
AMC Provider
out
2.rlal Diluter pH- Each lot of Failed sampie to be
I testing Buffer & diluter discarded recorded and
Page 36 of 75
prepared details to be given in
quarterly report.
I
Time
Page 37 of 75
Failed sample to be discarded, recorded and details to be
given in quarterly report
Note :
Bulls with more than 3O7o ejaculates being discarded in a month
may be subjected to additional tests at the discretion of the Oflicer In-
charge.
carriea out
I I
to be Failed samples
I
completed to be discarded,
in quarter) recorded and
I
details to be
I
given in
quarterly report
Page 38 of 75
of Semen cuE Post- (random) SOP be discarded,
Quality thaw samples in recorded artd
under motility storage for details to be
Long more than given in
I
(and
I
I
thereafter
tt
I
every six
I
tt months)
subjected
to PTM &
I
Incubation
i
I
test as
I
above
I
I
least twice
I
I
annually)
Page 39 of75
3.3 Use of Equipments for Advance qualitv Check: (Vlherever
laboratory is equipped)
taken I
decided by Sperm
I
station/ QCO
I I
Page 40 of75
4.O Feedback & Fertility Testins:
Page 4l of 75
identification by the respective SS as described in earlier section to
enable on-line reporting and real time data capture of semen production
and artilicial insemination.
The semen stations shall use suitable software to record data pertaining
to various activities and also should have online facility for the same. The
software should be able to identify and trace the bulls and their
ejaculates, production, storage and dispatch of semen etc.
f) Record of all quality tests for neat and frozen semen samples shall be
maintained.
Page 42 of 75
11 (Kl Semen Storage
To avoid accidental spread of diseases, tJ.e semen station shall follow the
procedure of preserving semen doses for at least 30 days (quarantine)
after production. Frozen semen doses produced at least 3O days prior to
the date of dispatch should only be supplied for AI.
12 Bio-security
Page 43 of 75
The risk of disease spread has grown manifold with increasing number
of bulls maintained at the semen production center. With the expected
higher risk, implementation of strict bio-security measures at the semen
stations assumes greater significance. Every semen station should have
a well defrned Bio-security protocol put in place across all its activities.
There shall be a designated Bio-security Officer (Veterinarian) at each
Sperm Station. The premises should be demarcated into high, medium
and low risk zones.
All the items to be washed shall be initially cleaned with running tap
water and soaked in warm neutral detergent for at least 30 minutes.
These items will then be thoroughly cleaned under running tap water
using a brush. Filling nozzles shall be cleaned with pressure using 20 ml
syringe. These materials shall be rinsed thoroughly with de-ionized/
distilled water (3 changes) to completely remove detergent residues and
other impurities. Appropriate procedure for sterilization of different
materials, recommended for use in the semen station follows.
Page 44 of 75
a) cone from the AV and water from AV jacket shall be removed before
washing.
b) Cones and AVs shall be cleaned thoroughly with a soft sponge brush
under running tap water and then soaked in warm neutral detergent
for about 3O minutes, followed by proper rinsing in warm and clean
water and then three times rinsing with double distilled water.
d) Finally, AVs hlled with water and covered with aluminium foil, at both
ends shall be stored overnight in an incubator at 45o C.
e) To achieve best cleaning effect, AVs sha11 be cleaned immediately after
use, preferably by non-spermicidal neutral detergent.
13.3 Glassware
a) The glassware shall be washed thoroughly with running tap water and
soaked in warm, non-spermicidal neutral detergent solution for about
30 minutes.
b) Using appropriate nylon brush, the glassware shall be cleaned and
rinsed with running tap water. The collection tubes shall be brushed
at least 3 times and thoroughly cleaned and rinsed with distilled
water.
c) Finally the glassware shall be rinsed three times with double distilled
water and allowed to dry by keeping them inverted on a blotting paper
or a drying stand made of SS/ plastic.
Page 45 of 75
d) The open end/s of the dried glassware shall be covered with
aluminium foil and sterilized in hot air oven at 160'C (holding) for one
hour or at 180'C for 30 minutes. One item should be wrapped with
newspaper and its mild charring will indicate proper sterilization.
Fresh triple glass distilled water or Ultra pure water shall be autoclaved
at 15 p.s.i. for 15 minutes and used for preparation of the dilutor.
13.6 Bulfer
Page 46 ol75
A bunch of clean filter papers of standard brand - No. 1 (thrashed to
remove dirt, if any) shall be wrapped in thick cotton cloth for sterilization
in an autoclave at 5 p.s.i. pressure for 20 minutes. Alternately. these can
be sterilized drv in suitablv sized dishes in hot air oven at 180
desree Centiqrade for 30 minutes.
14 Summary of Sterilization
Table:6 Autoclave
Sr.No. Item Pressure Time
I
(p.s.i.l (Min.f
1 Artificial Vagina 5 20
2. Piastic Tips 5 20
3 Filter Papers 5 ci
4 Bull Apron 5 20
wares
6. Bacteriological Media 15 15
7. Distilled Water 15 15
8. Surgical Equipment 10 10
I I
(min.)
Page 47 of 75
1 Glass wares 1600C/ 1800C 60l30
2. Filling Nozzles 1600C/ 1B0oC 60l30
cf AV Steriliser
wherever Autoclave is not used, AVs and rubber cones shan be sterilised
using AV sterilizer. After sterilizer starts boiling, 30 minutes vapour
sterilisation shall be do
d) Validation of Sterilization
Chem icals
The chemicals of only highest purity of either, Analltical
Reagent (AR) or
Guaralteed Reagent (GRf, from reputed manufacturing companies
shall be
used' whenever a new chemical is to be introduced in the routine
process, it
is recommended to examine the post-thaw revival rates after
conducting a
few spilt ejaculate trials (maintaining a controt) with the
new chemical.
Assay of chemicals shall be >99%o, having iess impurities.
Straws
Page 48 of 75
and sealing machine and the machine should be run to see the sealing
quality of the straws. In case of any foul smell, it should be presumed
that the straws are manufactured from poor plastic which could be toxic
to the spermatozoa and can even result in reduced motitity on long
storage.
2. The factory plug should not be loose. The factory seal should be
impenetrable and the seal formed should be homogeneous and compact.
3. The straws should be intact (without cracks / dents, etc.) and should not
get damaged during filling / sealing and after fteezing thawing.
/
4. The movement of straws along the printing machine should be free and
print should be clear and sharp. print should not fade as a result of
freezing and subsequent thawing.
5. The use of dark coloured straws should be avoided, as they are not
transparent enough. Due to this, it is diffrcult to distinguish between
lilled / semi-filled straws.
Page 49 of 75
16 Manpower Requiremeat for semen production
Tabie: 8
Page 50 of 75
Officer In-charge 1 1 1 1
QCO/QAO 1 1 1 1 1
Agriculture 1 1 1 1 1
Officer
Data Mgmt. 1 1 1 1
Ofhcer
Accts. & Adm 1 1 t-2 1-2
Oflicer I
I
Office Assistant 1 2 o
5 6-7
Livestock I 2 3 4 5
Assistant I
Agri. Assistant
I
Lab Technician I
1 2 3-4 I s-6 8-10
Vehicle/Tractor 1 2 3 4 5
Driver I I
Lab Artendant a
I
2 3-5 7-8 10-t2
Page 51 of 75
recruiting additional manpower. After recruitment, all new persons shali be
trained at any of the recognized institutes. Once trained, they shall continue
to work in the semen station at least for five years.
Page 52 of 75
Annexure- 1
I
maintaining their health status.
Bull Calf A male cattle or buffalo which has not yet
reached sexual maturity.
Known Animals originating from a semen station or
health status rearing station that is strictly complying with the
guidelines mentioned in the MSP.
MSP diseases MSP diseases are the set of diseases - the
I
causative organism of which should not be
present in the semen - or preferably in the bu1l.
I
Page 53 of 75
the rearing station to maintain their health
status.
Semen A farm along with semen processing facilities
station where adult bulls are housed for semen
collection and processing. A series of ciinical and
laboratory examinations, vaccinations and
medications etc. are undertaken during the stay
of bulls in the semen station to maintain their
hea,lth status.
Unkaown Animals originating fro m village or farm where all
health status the animals of the farm or the viliage have not
been tested against the MSp diseases
Page 54 of 75
Annexure- 2
oflicers of
Brucellosis ELISA Serum CDDL/RDDL/
I
NDDB/ State
Veterinary
I
Universities
TB- DTH- Intra-dermal on Semen station/
Tuberculin PPD the bull CDDL/RDDL/
NDDB/ State
Veterinary
I
Universities
JD- DTH- Johnin Intra-dermal on Semen station/
PPD the bull CDDL/RDDL/
NDDB / State
I
Veterinary I
I Universities I
Universities
Bovine Genital Agent Preputial CDDL/RDDL/
Campylobacterio identification washings NDDB/ State
sis Veterinary
I
Universities
FMD ELISA Serum PD-FMD,
I
Mukteshwar
I
and its
I I
I
laboratories/
Page 55 of75
NDDB /Stare
Veterinary
I
I
I
I
Universities
IBR ELISA Serum for CDDL/RDDL/
Real time- PCR ELrSA (9 NDDB/ State
months age) Veterinary
Semen for RT- Universities
PCR
BVD ELISA 2 times Serum CDDL/RDDL/
at 30 days NDDB/ Stare
I
interval I
Veterinary
(RT-PCR up to 6 I
Universities
I
I
months age)
Page 56 of 75
Annexure- 3A
Quarantine Guidelines
Page 57 of 75
B. Quarantine of adult bulls of known health status
Quarantine Minimum 30 days or long enough to a11ow at least
period one test for all MSP diseases
Shifting of Within 30 days of the last negative test
bulls from
the
quarantine
I
exterded
quarantine
Page 58 of75
Annexure- 3C
Page 59 of 75
Annexure- 3D
Page 60 of 75
last 30 days ofthe extended quarantine.
Action on Same as in Annexure- 3A
finding a
positive
during
extended
quarantine
Page 6l of 75
Annexure- 4
Page 62 of 75
Action to be Immediate isolation and removal from herd (within
taken on 2 days)
Positive
animal
Negative herd Six monthly (t 1 week) testing after last whole herd
I
affected animal.
I
the testing should establish that all animals in the
herd have been negative for the last 6 months
beginning from 6 months after culling the last
affected animal.
Page 63 of 75
Annexure- 5
Page 64 of 75
taken on 2 days)
Positive
anilnal
Negative herd Six monthly (t 1 week) testing after last whole herd
testing negative testing.
Page 65 of 75
Annexure- 6
Sample Serum
required
I
Negative herd Six monthly (t 1 week) testing after last whole herd I
Page 66 of 75
Annexure- 7
I
Bovine Genital A11 alimals are negative on two consecutive
Campylobacterio annual testing.
sis free herd
Page 61 of 15
Annexure- 8
Frozen semen Destroy fro zen semen doses of the positive animal
doses of the since last negative test. I
positive
animal
Page 68 of 75
Annexure- 9
Page 69 of 75
Annexure- 10
Name of the test Enzyrne Linked Immuno absorbent Assay (ELISA) for
detection of antigen (Ag-ELISA)/real time pCR (rt
PCR)
Sample Serum
Induction of new Test th e animai for Persistent Infection (pI) by testing
animals into two times at al interval of at least 30 days by Ag-
herd/semen ELISA. Test by rt-PCR instead ofAg-ELISA for
statious animals up to 6 months of age.
If the animal is positive on both the tests, the arlimal
is considered positive for pl. Only pI negative animals
shall be inducted.
Action to be taken Immediately isolate and cu11
for PI positive
animals
Semen dosses of PI Destroy by incineration of frozen semen doses of the
positive animals PI positive bulls.
Bulls at the semen (i) Test all the b ulls for PI (if not already tested for pl
stations status) by testing two times at an interval of at least
30 days. If the bull is found positive on both the
tests, then the bu1l is considered positive for pI. All pI
positive bulls shall be culled immediately.
(ii) Test all new bulls entering the semen station for
PL Only PI negative bulls should enter the semen
station.
Page 70 of 75
Annexure- 11
taken
burning.
Action to be
Isolate the affected bull immediately
taken on FMD
Affected bull is treated and rested for 90 days
infected
after recovery from clinical symptoms.
animal
a No semen collection from any infected animal
during the infection and up to 3 months after
last case has recovered in the farm.
Semen Sale lf frozen semen sale is from the same campus of the
SS where FMD is recorded, suspend semen sale till
30 days after the last case has recovered.
Page 7l of 75
the perimeter towards the focus.
Page 72 of 75
Annexure - 12
Feeding crowing aud Mature Bulls
Daily nutrient requirements of growing and mature bulls *
Growing bulls
l4
200 750 5.7 530 3.4
tt
18
tl 10 8
350 7so I
9.3 760 5.2 24 18 15
Page 73 of 75
700 10.9 I
830 6.t o< 19 I
30
I
I
bulls
150 2 o 8- 10
400 a) 2 I 3 ad lib.
b) 2.5 a
ad lib.
b) 3 2-4 ad lib.
b) 3 2-4 ad lib.
I
b) 2 2-4 ad lib.
600 -do-
Page 74 of 75
700 do-----------------
Calf Green
starter c.F. B.P.F. fodder Hay
DM o/o oo 90 90 20-25 90
I
Page 75 of 75