Minimum Standards
for
Production of Frozen Semen
April' 2005
CENTRAL MONITORING UNIT
Department of Animal Husbandry & Dairying
Ministry of Agriculture, Government of India
CONTENTS
Particulars
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
10 (A).
10(B).
10 (C).
10(D).
10(E).
10(F).
10(G).
10(H).
10(1).
10(J).
10(K).
11.
12.
13.
14.
Standards for Genetic Merit of Breeding Bulls
Physical Examination
Quarantine
Testing of Bulls
Vaccination Schedule
Culling of Bulls and Semen Doses due to Specific Diseases
Housing
Management of Bulls
Semen Collection
Handling of Semen, Processing & Freezing
Premises
Equipment
Personnel Hygiene
Diluents
Evaluation & Processing
Colour Specifications
Printing of Straws
Post-thaw Motility
Quality Checks for Frozen Semen
Information System
Semen Storage
Cleaning and Sterilisation of Laboratory Equipment
Summary of Sterilisation
Quality Control of Consumables
Manpower Requirement for Semen Production
Page
1
2
2
3
3
3
4
4
5
6
6
8
9
9
9
10
11
11
11
12
12
13
15
15
16
Annex.
_
0
%
"
I.
II.
III.
IV.
V.
VI.
VII.
VIII.
IX.
X.
XI.
Drawing
Tuberculosis Management
Jhone's Disease (JD) Management
Brucellosis Management
Infectious Bovine Rhinotracheitis (IBR) Management
Bovine Genital Campylobacteriosis Management
Bovine Trichomoniasis Management
Foot and Mouth Disease Management
Feeding Growing and Mature Bulls
List of Laboratory Equipment for Frozen Semen Stations
Qualifications Recommended for Semen Station Personnel
Job responsibilities of the QCO
Plan of a Model Semen Station
18
19
20
21
22
23
24
25
27
29
30
32-33
MINIMUM STANDARDS FOR PRODUCTION OF FROZEN SEMEN
Artificial Insemination with frozen semen has been proved to be the best tool world
wide for genetic improvement through dissemination of superior germplasm. This
objective can be achieved only if the frozen semen used in Al programme conforms
to the quality standards. For production and distribution of quality semen, it is most
important that the bulls used in Al programme satisfy pedigree norms, bulls are
disease free and semen is harvested and processed in accordance with the standard
protocols. The least protocols required for production of quality semen are covered in
this manual. Failure to observe these guidelines could lead to production of poor
quality semen making it unfit for distribution to AI centres.
The minimum pedigree standards for bulls in the Al programme are given in the
following table.
1. Standards for Genetic Merit of Breeding Bulls
Breed
Pure HF
Pure Jersey
Sahiwal
Red Sindhi
Gir
Kankrej
Tharparker
Hariyana
Rathi
Ongole
Deoni
Khillar
Dangi
Amritmahal
HFCross-F2
Jersey CB- F2
Sunandini
Murrah
Mehsana
Nil! Rav!
Jaffrabadi
Surti
Dam's Lactation yield (K
Imported
Farm bom
(minimum)
First
Best
4500
3000
2400
2000
2400
2000
2000
1600
1600
1100
300
380
400
400
4000
2800
2600
2400
2400
2400
2800
1600
5600
3750
3000
2500
3000
2500
2500
2000
2000
1600
1000
500
530
500
5000
3500
3000
3000
3000
3000
3500
2000
9000
6000
—
—
—
_
—
— .
—
—
—
—
—
—
~
—
—
—
—
—r
--------
gs)
Fat %
3.5
5
4
4.5
4.5
4.5
4
4
4
4
4
4
4
4
4
4.5
3.5
7
7
7
8
7
Sire's daughters' averages
of the bulls
Min number Average First
Reliability (%)
Lactn yield
of records
(Kgs.)
30
30
20
20
20
20
15
15
20
_
_
_
—
_
30
30
30
30
30
30
30
30
4000
2500
1500
1500
1500
1400
1400
1400
1500
—
. —
—
—
—
2500
2000
3500
1800
1800
1800
2000
1600
85
85
80
80
80
80
75
75
80
_
—
—
—
—
85
85
85
85
85
85
85
85
Dam's milk yield for F1 crosses will be as that of the indigenous dam's i.e. Gir,
Sahiwal, Kankrej, Red Sindhi, etc.
Minimum Standards
Average lactation yield (305 days) of HF and Jersey for the year 2002 for th e
following countries available from ICAR booklet are as below:
Breeds
New Zealand
HF
Jersey
3361
NA
Australia
6027
4381
Canada
8534
6048
US
9447
4619
France
Israel
6608
NA
10463
NA
The above information would be updated every year for information of all concerned
so that this data can be used during import of bulls.
2. Physical Examination
Before inducting new bulls in the semen station, a thorough physical examination
shall be conducted by an accredited Official / Veterinarian to ensure that the bulls do
not display clinical symptom(s) of any infection or any contagious diseases. Prior to
introduction of new bulls to the sperm station, breeding soundness
examination shall also be carried out. Standards for scrotal circumference and
weight gain index for various breeds shall be fixed by initiating age wise
(monthly) recording of scrotal circumference and body weight, by the semen
stations.
Before introducing to semen station, all bulls shall be karyotyped to rule out any
chromosomal defects.
Specific tests may also be conducted for genetically transmitted diseases like Bovine
Leukocyte Adhesion Deficiency (BLAD), Cttrullinemia and Deficiency of Uridine
Monophosphate Synthase (DUMPS) for HF and HF crosses.
3. Quarantine
A minimum quarantine period of 60 days is compulsory before bringing new bulls in a
semen station (incase of bulls are brought from known sources, 30 days quarantine
is sufficient). This period shall be divided in two periods of 30 days during which a
series of compulsory examinations and tests are carried out. Only after favorable
results from the health control point, the bull shall be admitted to the semen station.
a). In the quarantine station new bull/s shall be housed for minimum of 60 days in
a place which is effectively separated from (preferably at a distance of 5 km)
the facilities occupied by resident bulls and all equipment used in handling,
feeding, watering and cleaning the new bulls shall not be shared with the
resident herd(s).
b). Each new bull in quarantine station will be tested against major contagious
diseases before its entry to resident herd e.g. TB, JD, Brucellosis, Infectious
Bovine Rhinotracheitis (IBR/IPV), Campylobacteriosis and Trichomoniasis. All
tests shall be done by an accredited agency or disease diagnostic laboratory.
c) If any bull is found positive for TB, JD and Brucellosis, the positive bull shall
be removed immediately without retesting. The remaining bulls shall be
further tested twice for that particular disease at an interval of 60 days. If any
bull is found positive, all animals in that batch shall be culled. As regards
animals positive for IBR, culling shall be made based on isolation of virus for
IBR-IPV.
Minimum Standards
d) During quarantine period, the bulls shall be vaccinated against FMD, HS, BQ,
Theileriosis and Anthrax. However, vaccinations against bacterial diseases
shall be done only if there is an outbreak or prevalence of a particular
disease.
Once the quarantine period is over, all bulls shall be introduced to the resident herd.
The resident herd would also go through periodical testing and vaccinations as per
the schedule listed in the manual.
4. Testing of Bulls
Testing protocols for bulls against Tuberculosis, Johne's disease, Brucellosis, IBR,
Campylobacteriosis and Trichomoniasis are given in Annex I to VI. As per OIE
guidelines, the breeding bulls should be free from above mentioned diseases.
Though Johne's disease is not a sexually transmitted disease but from the herd
health point of view, bulls found positive should be removed and therefore it has
been included in the MSP.
5. Vaccination Schedule
The bulls shall be vaccinated against FMD, HS, BQ and Anthrax. However,
vaccinations against bacterial diseases shall be done only if there is an outbreak or
prevalence of a particular disease.
Theileriosis - to be vaccinated once in lifetime in exotic and crossbred bulls.
To reduce lay off time, the bulls shall be vaccinated on the rest day or the day after
completing semen collection. Sexual rest may not be required unless otherwise
febrile condition is noticed.
The semen station shall arrange for carrying out ring vaccinations for all animals
against FMD, HS and BQ within a radius of 10 km around the semen station.
Vaccinations against HS and BQ shall be carried out in the areas having incidence of
these diseases.
6. Culling of Bulls and Semen Doses due to Specific Diseases
Bulls
Diseases
FMD
Retain
Brucellosis
TB
JD
IBR/IPV
Castrate & remove
Remove
Remove
Remove based on
specific symptoms
Treat and retain
Treat and retain
Campylobacteriosis
Trichomoniasis
Minimum Standards
Semen doses
Last one month doses to be discarded, refer
Annex- VII
To be discarded since the last negative test
To be discarded since the last negative test
To be discarded since the last negative test
To be discarded since the last negative test
To be discarded since the last negative test
To be discarded since the last negative test
The semen station must remove bulls (within 48 hours) which are positive for above
diseases, poor libido, poor semen quality, incurable lameness, etc. Besides, the
semen station shall also cull those bulls which have completed six years of
productive period or 2 lakh semen doses, whichever is achieved earlier.
7. Housing
Bull sheds shall have spacious individual pens with adequate loafing area, manger
and water trough with access to drink water all time. Provision for adequate shade
around the bull shed shall be given priority. The roof shall be made of asbestos or
suitable materials. During summer cooling system with sprinklers and fan is required
particularly for the buffaloes and exotic bulls. Disinfectants like formalin or phenyl
based compounds shall not be used in the bull sheds. Alternatively compounds
containing Gluteraldehyde shall be used. Weekly spraying of Sodium Bicarbonate
(4%) solution shall also be practiced. The floor should be sterilized at least once a
year by a blowlamp or by burning straws. At one comer of the farm there shall be
isolation shed for separating ailing / sick bull for treatment. Bulls once diagnosed
suffering from infected disease, shall be removed immediately from semen station for
safety of other bulls.
8. Management of Bulls
The objective of daily care of bulls is to ensure a satisfactory state of cleanliness. For
proper management of bulls the following points shall be considered:
a) The bulls shall be kept under hygienic conditions at all times.
b) The coat of the bulls shall be kept clean and generally short. The hooves shall
be regularly trimmed.
c) The length of the tuft of hairs at the preputial orifice, which is invariably soiled,
shall be cut to about 2 cm. The hair would not be removed altogether,
because of its protective role. If cut too short, it may cause irritation of the
preputial mucosa.
d) Bulls shall be brushed and groomed regularly, and where necessary special
attention shall be given to the underside of the abdomen, a day prior to
semen collection. Also, prepuce shall be washed with sterile normal saline
solution.
e) In the event of obvious soiling, careful cleaning of the preputial orifice and the
adjoining areas with soap or a detergent is recommended; followed by
thorough rinsing and drying.
f)
Scientific feeding schedule shall be followed for the bulls. A general guideline
is attached as Annexure - VIII. Semen station shall carryout routine quality
analysis of feed and fodder for arriving at a balanced ration.
Minimum Standards
9. Semen Collection
a) Ideally, the floor of the collection yard shall be made of concrete layer at a
depth of one foot from the ground level. Mixture of sand and limestone shall
be used to fill up to ground level and pressed firmly. If it is not possible to
renovate the entire collection arena, at least the 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 AV / semen samples.
b) On the day of collection, before taking semen collection, the bulls shall be
properly washed and cleaned. After the bulls are brought to collection arena,
before taking collection, the prepuce shall be cleaned externally with a
sterilized napkin soaked in normal saline to remove any sand or dust
particles. For each bull a separate napkin shall be used.
c) The person responsible to carry out preputial wash must use disposable
gloves and separate sterilized nozzle for each bull to avoid transmission of
IBR infection from one bull to another.
d) The bulls shall be sexually prepared by giving two / three false mounts
followed by two minutes restraint and the total time limit should be not more
than 12 minutes. The duration of each false mount shall be for 1 to 2 minutes.
e) Sterilized bull aprons shall be used to avoid penis touching hindquarter of the
dummy.
f) Preferably veterinarians shall take semen collection. If semen is collected
by staff, a veterinarian shall remain present to supervise the collection
process. While taking collection it shall be ensured that AV is not thrusted on
penis of bull, instead penis should be guided to AV.
g) Before every collection, the semen collector shall either wash his hands with
0.1% Savlon solution or use disposable gloves or do both. The semen
collector shall not touch the penis.
h) Immediately after collection, the AVs shall be thoroughly cleaned by nonspermicidal neutral detergent. Separate AVs shall be used for each mounting.
The AV shall be changed even if the bull has inserted its penis without
successful ejaculation. The same AV shall not be used twice. The AVs shall
always be kept inverted and the collection tube shall be covered with felt /
water jacket (plastic bottle filled with warm water at 34° C) to avoid cold
shock. The open end of sterilized AVs shall be covered with aluminum foil,
which would be removed at the time when bulls are ready for giving semen.
i) Preferably 10° size AVs shall be used for cattle and 8 to 9" size for buffaloes
to ensure semen is ejaculated in cone. For buffaloes goat AVs can also be
used. The cone shall be of top quality Neoprene rubber.
Minimum Standards
j) Use of lubricant shall be avoided. If it is extremely essential to use lubricant,
separate sterilized glass rods shall be used for smearing K-Y Jelly on each
AV.
k) The AV shall not to be shaken after ejaculation; otherwise lubricant and
debris may mix with the semen samples.
I) The entry of visitors/ staff / labourers shall be strictly prohibited in the
collection arena at the time of semen collection and inside the semen
laboratory.
m) Protective clothing (barn coat) and gumboots shall be used by the
Veterinarians and personnel while taking semen collection. Gumboots and
bam coat should be washed daily immediately after completion of semen
collection work.
n) Semen must be obtained from a bull having normal libido. While taking two
ejaculations the semen station shall keep a gap of 10 to 12 minutes between
two ejaculates, depending upon the refractory period of the bull. (To harvest
more quantity of semen, interval between two ejaculates shall be kept for 40
to 45 minutes. After taking first ejaculate, the bull shall be taken back to the
shed for feeding and watering and again brought back for taking second
ejaculate. However, care shall be taken to use a fresh & sterilised apron
during second ejaculate)
o) Semen stations must follow the norm of two ejaculates per collection and two
collections per bull per week for taking annually at least 90 collections and
180 ejaculates from each adult bull.
10. Handling of semen, processing & freezing
10 (A) Premises
a)
Sufficient trees shall be planted and lawns prepared around the semen
station to reduce dust.
b)
The ceiling and walls of the laboratory shall be made up of non-porous
materials. All cracks and crevices shall be sealed to control pests and
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 lab.
d)
Lab windows shall be made of single sheet glass with fixed aluminium
frame. The glass panes shall be plastered with sun control films to avoid
direct sunlight. The doors shall be kept closed, especially during dilutor
preparation and semen processing.
e)
Intake areas of the air conditioners shall be -disinfected weekly and if
possible, split type air conditioners with remote temperature control
mechanism should be installed to maintain the room temperature at
Minimum Standards
20°C. The number of ACs to be fixed to sustain this temperature
shall depend on the size of the processing room. Maintaining this
temperature is most important to achieve the best results when single
step dilution method is followed for freezing semen. Alternatively, HEPA
filters shall be installed near intake area of air conditioners. The flow of air
from AC must not be towards the front side of the Laminar Air Flow Unit.
Adequate number of dial thermometers shall be kept in a few places in the
laboratory to check the room temperature.
f)
Sink drains shall be decontaminated routinely with a disinfectant.
Preferably, the sink shall not be placed in the semen processing room.
g)
The floors shall be preferably made up of vitrified tiles. Floors and
horizontal surfaces shall be cleaned and mopped with a disinfectant
solution, as dirt and dust, which settle on these surfaces, are the main
sources of contamination.
h)
Unwanted furniture, equipment and materials shall not be kept in the
laboratory as they only provide additional area for dust and spores to
collect.
i)
Appropriate no. of germicidal UV lights (2470 A) in respect of area
of laboratory, laminar airflow unit, apron and laboratory footwear
cabinet shall be fixed with a common operating switch outside the
laboratory. These lights shall be kept 'on' at least for 8 hours prior to
commencement of work in the laboratory and shall be switched off before
beginning work The date of installation of the UV lights shall be noted
to facilitate replacement as the life of UV tube is of 2000 hours. A
logbook should be maintained for timely replacement of UV lights.
j)
Laboratory shall be fumigated weekly for 2 hours with 12 ml of 37%
formaldehyde solution in 100 ml water per cubic meter at weekends, using
humidifier. After fumigation, switch on the exhaust fans for clearing
formalin fumes. However, this should be supported by monitoring of lab
environment by bacterial load test. The bacterial ioad shall be measured
every week to monitor pollution of the laboratory atmosphere.
k)
(Mixing formalin and water with potassium permanganate crystals*.
*WARNING: the correct relative concentration of these two components is
essential to avoid a violent reaction. It is therefore recommended that this
method is NOT used. (Reference; Bid-safety Unit, Health and Safety
Department, University of Edinburgh, August 2003)
I)
The work platform, the parts of equipment and other items to be
handled during processing of semen, shall be cleaned with 70% alcohol
or Glutaril (Qualigen). It is advisable to repeat cleaning schedule after
completing processing of semen.
m)
Clean laboratory footwear, apron, hand gloves, mask and caps shall be
compulsorily put on while working in the laboratory.
n)
Eating, drinking, smoking, etc. shall be prohibited in the laboratory and
unnecessary conversation should be discouraged. Besides, entry of
persons shall be strictly restricted.
Minimum Standards
o)
Long exposure of semen to ultraviolet rays, visible light in direct
sunlight and white florescent light causes chromosomal damage and
hence direct exposure to such sources of light shall be avoided. Hence,
there shall be provision for indirect or diffused lighting inside the semen
processing room. Care shall also be taken not to switch on tube lights in
CH cabinet and laminar airflow unit.
10(B) Equipment
a)
The exteriors of all equipment and furniture shall be cleaned weekly. The
equipment shall be kept covered by plastic covers when not in use.
b)
The pre-filter of Laminar Airflow unit shall be cleaned weekly. Routine
servicing and OOP testing twice a year will ensure efficiency of HEPA
filters. Alternatively culture plate test shall be carried out at frequent
interval to assess bacterial load of the air passing through the filters.
c)
Digital photometer / Computer aided Spectrophotometer shall be validated
with Haemocytometer readings for sperm concentration twice a year
separately for cattle and buffalo (10 samples each).
d)
The automatic semen straw filling and sealing machine shall be
thoroughly cleaned, immediately after use.
e)
The microscope lens shall be gently cleaned daily with a piece of cotton
soaked in a mixture of ethyl and methyl alcohol (1:1).
f)
The bio-freezer shall be defrosted and thoroughly cleaned and dried,
immediately after use.
g)
Incubators to maintain artificial vagina shall be cleaned and
disinfected daily with 70% alcohol.
h)
Single distilled water shall be used in autoclave and thermocontrolled water bath. The water bath shall be cleaned and filled with
single distilled water on a regular basis.
i)
The thermometer kept immersed in water bath shall be cleaned
daily to have precise temperature reading.
j)
The Liquid Nitrogen containers returning / received from foreign
countries and contagious disease prone areas shall be disinfected
thoroughly with 4% soda solution and finally with 1 to 4% formaldehyde.
Excess liquid nitrogen available as a result of filling the containers with
canisters and goblets, shall never be brought inside the semen
processing room.
k)
The refrigerator meant for storing eggs, antibiotics and buffer shall not
be used for storing vaccines and other materials. All such materials shall
be stored at a place away from semen lab. The refrigerator used for
storing eggs, etc. shall be sterilized every week using alcohol swab.
Minimum Standards
8
10 (C) Personnel Hygiene
Clothing, skin and hair of laboratory personnel are the sources of
the contamination. Hence all should wear laboratory aprons and footwear all
the time while they are in the laboratory. Hands shall be washed with soap
and water and rinsed with 70% alcohol, before commencing work in the
laboratory. The bull attendants must undergo test for TB every year.
Restricted entry inside the semen processing room and freezing room shall
be strictly adhered to.
10 (D) Diluents
a) All disposable and reusable supplies coming in contact with the semen and
dilutor must be sterile and devoid of toxins and pyrogens.
b) Prolonged storage of purified water is not recommended because water purity
deteriorates progressively over a period of time as heavy metals leach from
some glass and plastic storage vials / containers.
c) Glass wares, collection tubes, etc. shall not be handled from their rim / mouth.
d) Pipetting shall be done away with, instead adjustable micropipettes and
disposable tips shall be used.
e) After adding all the components of buffer viz. TRIS, Citric Acid, Glycerol and
Fructose in double, preferably triple distilled water, it should be again
sterilized. If buffer is prepared on the previous day and stored in the
refrigerator then antibiotics are to be added on the next day morning after
warming it at 34°C.
f) Antibiotics in diluents: Combination of antibiotics, in diluents, which can
control Mycoplasma like Tylosin, Lincospectin and Gentamycin may be used.
Alternatively, a combination of Penicillin and Streptomycin shall be used.
g) The eggs used for making dilutor must be fresh. The eggs shall be stored in
refrigerator after wiping with dry cotton. Just before preparation of dilutor,
eggs shall be wiped with 70% alcohol. To avoid Mycoplasma infection, eggs
shall be purchased from known sources.
h) The required quantity of yolk shall be separated from albumin on sterile
(autoclaved) standard filter papers (Whatman No. 1/ Borosil) and yolk
membrane shall be punctured using sterile glass rod or Pasteur pipette or
sterile straws under the Laminar Flow Unit. Only fresh semen extender/dilutor
shall be used because changes in the pH of stored extender are considered
to be responsible for the deterioration of some nutrient components. Day old
extender shall not be used.
10 (E) Evaluation & Processing
a) As soon as the neat semen is received, it shall be kept in a thermo-controlled
water bath at 34° C under Laminar Unit, after recording the volume of semen.
Minimum Standards
b) The tube containing the freshly collected semen should be capped with
aluminum foil as soon as it is placed in the pass box before transferring to the
laboratory. The collection tube shall be kept capped until processed.
c) After examination of sperm concentration and initial motility, semen samples
shall be primarily diluted with dilutor maintained at 34° C. In the case of
buffalo semen, examination of mass activity is mandatory because semen
samples with zero mass activity may not give better result and usually
discarded after freezing. Alternatively, a small drop of fresh semen shall be
examined under a cover slip to check motility and to assess the semen
samples.
d) Sperm concentration shall be checked preferably by a digital photometer with
auto dilutor, manufactured by a reputed company. The photometer shall be
calibrated separately taking 10 readings each for cattle and buffalo semen, at
least once in six months, with haemocytometer readings. Semen samples
showing less than 500 million / ml sperm concentration shall be discarded.
e) Semen samples selected for freezing should have minimum 70% initial
motility. Final dilution of semen keeping a minimum of 20 million spermatozoa
per dose shall be done in appropriate flasks with the dilutor maintained at 34°
C.
f)
Filling and sealing of semen shall be done under Laminar Flow Unit using
sterile straws, filling nozzles and fresh rubber tubings. Rubber tubings shall
be used once only. Reuse of rubber tubes is not recommended. Considering
the advantages the French Mini Straws has over French Medium straws, the
semen stations shall use French Mini straws.
g) Unused straws shall be repacked (air-tight) under Laminar Unit before
storage.
h) Immediately after use, all the glass wares, rubber wares, plastic tips and other
reusables shall be immersed in neutral detergent solution (to be kept in a
plastic tub near the Laminar Unit).
10(F) Colour Specifications :
All semen stations shall follow the following colour codes :
1.
2.
3.
4.
5.
6.
7.
If any
Jersey
Yellow
Holstein
Pink
Indigenous
Orange
HF Crossbred
Pistachio Green (light green)
Jersey Crossbred
Salmon
Sunandini
Blue
Buffalo
Grey
of above colour is not available then transparent colour is used.
Minimum Standards
10
10 (G) Printing of Straws
Information regarding bull number, breed, name of the organization, year, batch
no. (as per the day of the year), etc. shall be printed on straws preferably after
their filling and sealing. After printing the ink gets instantly dried. If filled straws
are printed and racked, the actual number of straws can be easily counted. While
printing and racking, the room temperature shall be maintained at 20 ° C.
AH semen stations shaii foiiow the following printing abbreviations:
Jersey
Holstein
HF Cross
Jersey Cross
Sunandini
Sahiwal
RedSindhi
Kankrej
Gir
Tharparker
Rathi
Haryana
Murrah Buffalo
Surti Buffalo
Jaffrabadi Buffalo
Mehsana
^-
JY
HF
CBHF
CBJY
SUN
SAH
RS
KANK
GIR
THAR
RATHI
HAR
MBF
SBF
JBF
MSNB
Farm No. / Name
Breed
Name of Institute
Batch No. / Date of Prodn.
10 (H) Post thaw motility
After freezing, the semen straws shall be stored in a separate container. Postthaw motility of semen should be examined at 24 hours. Differences in
observations shall be updated and recorded for the purpose of accepting a
particular batch of semen doses. Whenever there is any doubt, post-thaw motility
shall be examined by two experienced persons. Preferably, the person involved
in evaluation of neat semen, shall not check the post thaw motility. For a
minimum concentration of 20 million per dose, minimum acceptable post thaw
motility shall be 50%. Semen doses below 50% progressive motility shall be
discarded.
10 (I) Quality Checks for frozen semen
This includes (i) Quarterly testing of random samples from each batch for
bacterial load using standard plate count (The standards for acceptable colony
forming unit in processed semen is 5000 CFU per ml as per OIE norm. If the
bacterial load exceeds the OIE limit, the semen doses are to be discarded.) (ii)
Hypo osmotic swelling test (HOST) - every day four to six samples (iii) Incubation
test - everyday four to six samples (iv) Acrosome integrity test by Giemsa
staining - for all bulls at least once in a quarter shall be mandatory. Alternatively
wet smear of semen shall be examined using DIC microscope (v) Percent Intact
Acrosome - all bulls to be covered once a quarter (vii) Sperm Concentration randomly two samples per week each for cattle and buffalo
Minimum Standards
11
Validation of photometer shall be done once in 6 months, at least 10 samples
each for cattle and buffalo. Neat semen shall be examined at an interval of every
six months for morphological abnormalities particularly for crossbred bulls.
Morphological examination of sperm of young bulls must be carried out (at least
six samples at weekly intervals) before introducing them in the herd. Semen
should not be used if the sample contains a total abnormality of more than 20%
and head and mid-piece abnormality ( alone) of 7%.
10 (J) Information System
a) The semen stations shall use a suitable software to record data pertaining to
various activities and also should have online facility for the same. The semen
stations producing more than one million doses may introduce software that
can identify and trace the bulls and their ejaculates, production, storage and
dispatch of semen (barcode system).
b) Volume of semen, density, motility, sperm concentration, dilution rate, total
extended volume, post-thaw motility (24 hrs), total number of doses
produced, etc. shall be maintained. Pre-freeze and post-thaw motility shall be
checked for new and problematic bulls.
c) Miscellaneous information regarding actual reason(s) for not donating semen,
undesired percentage of gross morphological defects, semen pH, presence of
dirt, dust, blood, pus, etc. in semen samples.
d) Details of semen supplied to various agencies including post-thaw motility at
the time of dispatch.
e) Fertility data of bulls, conception rate, records of the progeny associated with
any genetic defect, percent male / female born, etc.
f)
Report on microbiological examination of semen samples.
g) Records of all quality tests for neat and frozen semen samples.
10 (K) Semen Storage
To avoid accidental spread of diseases, the semen station shall follow the
procedure of preserving semen doses for at least 30 days after production.
Semen doses produced before 30 days from the date of dispatch shall only be
supplied for field use.
After checking post-thaw motility, if found acceptable, frozen semen doses
would be kept in temporary storage for 7 days. After the temporary storage the
semen goblets shall be transferred to the bulk storage containers with proper
recording of position in the canisters. After each dispatch, records redefining the
position of remaining doses shall be updated.
Liquid Nitrogen shall be replenished at regular intervals depending on the liquid
nitrogen evaporation rate of the container.
Minimum Standards
12
11 Cleaning and Sterilisation
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 water (5 to 7 changes) to completely remove
detergent residues and other impurities. Appropriate procedure for sterilization of
different materials, used in the semen station, is given below:
1. Laboratory and other areas
a. Materials
1.
2.
3.
4.
Fumigator: Aerosol formation disinfector
Formaldehyde Solution 37 % w/v
Lab dimensions
Lab area in cubic meters
b. Procedure
1 . Review of the microbial count indicates the necessity of fumigation
2. Calculate the area in cubic meter, which is to be fumigated
3. Calculate the formaldehyde solution using dosage 12 ml of 37% w/v of the
formaldehyde solution in 100 ml water per M3
4. Decide the no. of fumigators to be used (based on area)
5. Put total quantity of formaldehyde solution in all the fumigators
6. See the doors and windows are sealed properly
7. Switch off the exhausting fans and other fans and switch all the fumigators
from outside switch
8. Evaporate full formaldehyde solution and switch off fumigators after
completing fumigation (about 2 hrs.)
9. Ensure that all the dampers of exhaust area, ducts are shut off during
fumigation
10. Switch on the exhaust fans / air handling systems after 2 hours of fumigation
11. Inspect the room for the absences of formalin fumes before the personal entry
12. Expose the settle plates and record the microbial count
1 3. Review the microbial count so as to establish the effectiveness of fumigation
c. Maintain a register to record observations on microbiological load in
semen processing lab.
2. Artificial Vagina (AV)
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 cleaner for about 30
minutes, followed by proper rinsing in warm and clean water and lastly rinsed
three times with double distilled water.
Minimum Standards
13
c) For sterilization, fully assembled AVs shall be autoclaved at 5 p.s.i. pressure
for 20 minutes. During sterilization the AV - valve shall be kept open.
Alternatively, use AV sterilizer (using double distilled water in the sterilizer) for
proper sterilization of AVs.
d) Finally the AVs shall be stored overnight in an incubator at 45° C.
e) To achieve best cleaning effect, AVs shall be cleaned immediately after use,
preferably by non-spermicidal neutral detergent.
3. Glasswares
a) The glasswares 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 glasswares 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 glasswares shall be rinsed three times with double distilled water
and allowed to dry by keeping them inverted on a blotting paper.
d) The open end/s of the dried glasswares shall be covered with aluminium foil
and sterilized in hot air oven at 160° C for 1 hour or at 180° C for 30 minutes.
4. Rubber wares
The washing and cleaning procedure of rubber wares is similar to that of
glasswares. Care shall be taken to clean the rubber wares with sponge brush
instead of nylon brush. Plastic tips shall be cleaned by water jet with force using a
syringe. Sterilization technique, however, differs owing to the thermo-sensitivity of
the rubber items. Thermo-sensitive rubber wares shall be packed and sealed in
specific polythene bags and sterilized in Ethylene Oxide gas sterilizer. Thermoresistant rubber wares shall be sterilized by autoclaving at 3 - 4 p.s.i. for 10
minutes. (The rubber tubing for semen filling shall not be reused).
5. Distilled Water
Triple glass distilled water or Milli-Q purified water shall be autoclaved at 15 p.s.i.
fop 45 ^minutes and used for preparation of the dilutor. The Triple glass distilled
water or Milli-Q purified water stored for more than 5 days shall not be used for
buffer preparation.
6. Buffer
Buffer shall be sterilized by autoclaving at 5 p.s.i. pressure for 20 minutes. After
autoclaving buffer shall be cooled and stored in refrigerator.
7. Bacteriological Media
It is to be autoclaved at 15 p.s.i. pressure for 15 minutes.
Minimum Standards
14
8. Filter Papers
A bunch of clean filter papers of standard brand like Whatman 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.
12 Summary of Sterilization
a) Autoclave
Sr.No.
1.
2.
3.
4.
5.
6.
7.
8.
9.
Item
Pressure
Time
(p.s.i.)
(Min.)
Artificial Vagina
5
20
Buffer
5
15/20
Plastic Tips
5
20
Filter Papers
5
20
Bull Apron
5
20
Thermo-resistant Rubber wares
3-4
10
Bacteriological Media
15
15
Distilled Water
15
15
Surgical Equipment
10
10
(The rubberwares can withstand above pressure and duration provided the quality is
good)
b) Hot Air Oven
Sr.No. Item
1.
2.
Time
(min.)
60/30
60/30
Temperature
Glass wares
Filling Nozzles
c) AV Steriliser
160°C/180°C
160° C/ 180° C
Wherever Autoclave is not used, AVs and rubber cones shall be sterilised using AV
sterilizer. After sterlizer starts boiling, 30 minutes vapour sterilisation shall be done.
d) Ethylene Oxide Gas Steriliser
The exposure time for gas sterilization is inversely proportional to the gas
concentration under which the equipment is exposed in the sterilizer. Commonly
used concentration of ethylene oxide is 900 mg per litre of cubic space for a period of
six hours. All items sterilized with gas must be aired adequately before use. A
minimum exposure of 72 hours at room temperature with adequate ventilation is
recommended.
13 Quality Control of Consumables
Chemicals
The chemicals of only highest purity of either, Analytical Reagent (AR) or Graded
Reagent (GR), from reputed manufacturing companies shall be used. Whenever a
new chemical is to be introduced in the routine process, it is recommended to
Minimum Standards
15
examine the post-thaw revival rates after conducting few spilt ejaculate trials
(maintaining a control) with the new chemical. Assay of chemicals shall be >99%,
having less impurities.
Straws
1. Straws manufactured by reputed companies are safer to use for production of
quality semen. While buying straws package volume and microbial load in
straws shall be checked randomly from the consignment. In addition, some
empty straws should be placed in auto filling & sealing machine and run the
machine for sealing the straws. In case 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 motility 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.) during and after freezing
/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. Not only is filling / racking inconvenient, it is also difficult to distinguish
between filled / semi-filled straws.
6. Movement of the factory plug should be free.
7. Straws should be routinely checked for microbial load.
Note: The semen stations should avoid purchase of consumables on lowest
quotation basis. To produce top quality semen, it is better to use AR /
GR consumables manufactured by reputed companies whose
products are reliable.
14 Manpower Requirement for semen production
Designation
QIC
vo
QCO
Lab Technician
Lab Attendant
Office Assistant
Bull Attendant
Minimum Standards
Up to 5 lakh
doses
1
1
1
.—
2
—
5-10 lakh
doses
1
10-25 lakh
doses
1
2
1
2
3
1
2
1
3-4
3-5
1
1 person per 7- 8 bulls
16
Above 25 lakh
doses
1
3
1
4-5
4-6
1
The manpower structure suggested above, is meant only for semen production. For
dispatch of semen, facility should be created preferably away from semen station and
operated by other person/s not responsible for semen production. The GOI /
Department of AH / Livestock Boards / NGO / Private agencies / Union and
Federation shall review the requirement of manpower position for each semen station
and finalise the staff structure for recruiting additional manpower. After recruitment,
all new persons shall be trained at any of the recognized institutes. Once trained,
they shall continue to work in the semen station at least for five years.
Refresher training / visit to other semen lab technical exposure of semen station
personnel working in the semen lab must be arranged compulsorily once in two to
three years at reputed institutions like SAG-Bidaj, CFSP&TI-Hessarghatta, KLDBMatupatty, etc. As semen production activity is an extremely technical work, frequent
transfer of personnel could be detrimental in maintaining the quality of semen.
Therefore, before considering transfer, in the interest carrying out good work it is
essential that proper replacement is identified at least six months in advance and is
trained in semen production technology.
Minimum Standards
17
Annex-1
Tuberculosis Management
Screening Test
details
Name
Delayed Hypersensitvity Single Intradermal test
Reagent
Reagents from
Testing at
Jovine tuberculin PPD
VRI, Izatnagar
Where animals are housed by
RDDL/CDDL/NDDB
As per OIE norms Negative : Increase in
skin thickness less than 2 mm & without
clinical signs viz. exudation, necrosis, pain,
inflammation of the lymphatic duct of that
region or the ymph node, 72 hours postinoculation. Inconclusive : Increase in skin
thickness more than 2mm & less than
4mm, absence of above clinical signs, 72
hours post-inoculation. Positive : Increase
in skin thickness 4 mm or more, or
presence of dinical signs viz. exudation,
necrosis, pain, inflammation of the
lymphatic duct of that region or the lymph
node, 72 hours post-inoculation.
Positive result criteria
Eligible animals
Frequency of
testing
Action on finding
a positive bull
Positive herd
Negative herd
Annual test is minimum. Six months (± 1
week) after last whole herd negative
testing, desirable.
Animal
Immediate isolation and elimination
Semen
Destroy semen doses since last negative
test
Herd found negative on two consecutive
tuberculin tests at an interval of 6 months,
the first being performed 6 months after
the slaughter of last affected animal
Tuberculosis free
herd (OIE)
Quarantine
All animals above 6 months of age, have
shown a negative result to at least two
tuberculin test carried out at an interval of 6
months
Minimum 60 days after culling of last
positive animal
Duration of quarantine
Test schedule
Minimum Standards
Minimum 90 days
Two tuberculin tests, minimum interval of
60 days between tests.
18
Annex - II
Johne's Disease (JD) Management
Screening test
Details
Name
Delayed Hypersensitivity test (Skin
test)
Reagent
Reagents from
Testing at
Johnin PPD
IVRI, Izatnagar
Where animals are housed by
RDDUCDDL/NDDB
Increase in skin thickness of over
4mm (discrete circumscribed
swelling), 72 hours post-inoculation
Positive result criteria
Eligible animals
Frequency of
testing
Action on
finding a
positive bull
Quarantine
Positive herd
All animals above 6 months
Minimum 60 days after culling of last
positive animal
Negative herd
Annual test is minimum. Six months
(± 1 week) after last whole herd
negative testing, desirable.
Animal
Semen
Immediate isolation and elimination
Destroy semen doses since last
negative test
Duration of quarantine
Minimum 90 days
Test schedule
Two Johnin tests, minimum interval of
60 days between tests.
Minimum Standards
19
Annex- III
Brucellosis Management
Screening Test
Name
ELISA, RBPT+CFT
Details
Sample
Reagents from
Testing at
Serum
CDDL / RDDL
• Ail above one year
• In females 14 days after calving or
abortion
Eligible animals
Frequency of
testing
Action on
finding a
positive bull
Positive herd
30 to 60 days after culling of last
positive animal
Negative herd
Exactly one year (± 1 week) after last
whole herd negative testing
Negative herd (optional)
Where the disease has been
maintaining a very low profile (less
thanl % positive) quarterly or six
monthly sample could be collected to
minimize losses
Animal
Immediate isolation and elimination
after castration
Destroy semen doses since last
negative test
Semen
Brucellosis free
herd (OIE)
Quarantine
CDDL/ RDDL/NDDB
Herd found negative on two
consecutive annual tests
Duration of quarantine
Test schedule
Additional
testing at sexual
maturity
Minimum Standards
Minimum 30 days
Two tests, Serum ELISA, interval of
30 days between tests. Only negative
animals to be allowed to mix with the
rest of the herd.
Serum ELISA before bulls are used
for semen collection and distribution
forAI
20
Annex- IV
Infectious Bovine Rhinotracheitis (IBR) Management
Screening
test details
Eligible
animals
Frequency of
testing
Name
Sample
Testing at
ELISA /SNT
Serum
CDDL / RDDUNDDB
All animals
Positive
Herd
Whole herd test, 30 to 60 days after culling of
positive animals. Six monthly, after the herd
become negative.
Exactly one year (± 1 week) after last whole
herd testing
Where the disease has been maintaining a low
profile (less than 5 % positive) quarterly or six
monthly sample could be collected to minimize
losses
Immediate isolation and elimination after
castration
Negative
herd
Action on
finding a
positive bull
Animal
Semen
Destroy semen doses since last negative test
IBR free herd
(OIE)
^
Quarantine
Duration of
quarantine
Testing
Whole herd tested negative on two consecutive
occasions at an interval of 2 to 12 months
between tests
Minimum 30 days
Additional
testing at
sexual
maturity
Minimum Standards
Two tests, Serum ELISA/ SNT, interval of 21
days between tests
Serum ELISA/SNT before bulls are used for
semen distribution in field Al programmes
21
Annex- V
Bovine Genital Campylobacteriosis Management
Screening test
details
Name
Bacterial isolation & identification
Sample
Testing at
Eligible animals
Preputial washing, semen
RDDL/CDDUNDDB
All male animals
Prevention
Annual sheath lavage
Frequency of
testing
Positive herd
30 days after culling of positive animal.
Negative herd
Exactly one year (± 1 week) after last whole
herd negative testing
Action on
finding a
positive bull
Animal
Treat the animals
Semen
Destroy semen doses since last negative
test
Quarantine
Duration of
quarantine
Test schedule
Minimum 30 days
One test if age is less than 6 months, else 3
consecutive tests at weekly intervals.
Minimum Standards
22
Annex - VI
Bovine Trichomoniasis Management
Screening test
details
Name
Agent isolation & identification
Sample
Testing at
Preputial washing
RDDL/CDDUNDDB
All male animals
Annual sheath lavage
Annual
Action on
finding a
positive animal
Animal
Treat the animals
Semen
Destroy the semen doses since last
negative test
Quarantine
Duration of quarantine
Test schedule
Minimum 30 days
One test, if age is less than 6 months,
else 3 consecutive tests at weekly
intervals.
Eligible animals
Prevention
Frequency of
testing
Additional
testing at sexual
maturity
Protozoa isolation before bulls are
used for semen distribution forAI
Minimum Standards
23
Annex-_V!l
Foot and Mouth Disease Management
Action during
FMD outbreak
Animal
solate diseased animals till recovery, do not
cull.
Semen
Semen from FMD infected bulls : Destroy
semen collected during one month
before onset of outbreak. Do not collect
semen from bulls during the outbreak and
three months after the last case of FMD
recovered in the farm.
Infected animals must be given 90 days
rest
Semen from healthy bulls maintained in
FMD infected farm:
Destroy semen collected during one month
before onset of outbreak. Do not collect
semen from bulls during the outbreak and
one months after the last case of FMD
recovered in the farm.
Semen could be used other than the above
mentioned periods, if there is no new case of
FMD develops during three months period
after last FMD case recovered in the farm.
Semen for export
Vaccination - as per manufacturer's
recommendations.
Quarantine
Vaccination
Test each batch by virus isolation and PCR
Oil vaccine
Minimum Standards
As per manufacturer's recommendations in
farm and 10 km around the farm
Test for seroconversion, by collecting serum
on the day of vaccination and 21 days later.
24
Annex-VIII
Feeding Growing and Mature Bulls Daily
nutrient requirements of growing and mature bulls
Body wt
(kg)
Growing bulls
100
150
200
250
300
350
400
450
500
550
600
gain/day DM/da c.P.(g) TON (kg) Ca(g)
(g) y (kg)
750
750
750
750
750
750
700
600
400
250
100
2.8
4.3
5.7
7
8.2
9.3
10.2
10.4
10
10
9.8
390
460
530
610
680
760
820
875
885
845
800
Maintenance of mature breeding bulls
8.3
640
500
600
700
-
9.6
10.9
735
830
VitA
P(g)
(1000 IU)
1.9
2.7
3.4
4
4.6
5.2
5.7
5.8
5.6
5.6
5.5
11
15
18
21
23
24
25
26
26
25
24
8
11
14
16
17
18
19
20
20
19
18
4
6
8
10
13
15
17
19
21
23
26
4.6
5.4
6.1
20
22
25
15
17
19
21
26
30
Daily ration for Bulls
Growing bulls
100
150
200
300
400
a)
500
600
b)
a)
b)
a)
b)
2
-
b)
Body wt.
(kg)
(kg)
-
2.5
3
3
-
2.5
-
2
.
C.F................B.P.F.
....... ..
(kg)
Minimum Standards
2
2
2
2
2.5
2.5
-
• -
Mature breeding bulls
500
a)
600
Calf starter
?nn
-
(kg)
. .. .. HO
_ ._
0.5
0
0.5
1
3
3
2-4
2-4
2-4
2-4
adljb_.
ad }ib.
ad lib.
ad lib.
ad lib.
id Kb,
ad lib.
2-4
2-4
ad lib.
ad Kb,
______
Hay
Green Fodder
. ............
rin ..............................
- ,- - ...
(kg)
(kg)
25
6-8
8-10
15
Note : 1)
Mineral mixture should be supplemented as follows :
50 g mineral mixture for bulls up to 200 kg body weight
70 g mineral mixture for bulls between 200 to 350 kg body
weight.
100 g mineral mixture for bulls above 350 kg body weight
2)
Fresh water should be made available 24 hrs.
Green fodder requirement of 10 mature bulls would be approx. 125 MT per year,
which can be grown in 1 hectare of land by intensive farming.
* Source: Ranjhan, S.K (1980). Animal nutrition & feeding practices in India,
' ______ 2nd Ed., p196-212 ____________
___
_________
Nutrients available in feed & fodder
Calf starter
DM%
CP%
TDN%
C.F.
B.P.F.
90
90
90
22-23
18-19
62-64
22-23
65-68
70
Minimum Standards
Green fodder
20-25
Hay
5-6
90
5-6
55-60
55
26
Annex - IX
List of laboratory equipment for Frozen semen stations (A).
Processing Lab
Sr.
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
Items
Phase contrast microscope with
Biotherm (Nikon/ Olympus)
DIC Microscope with bio-therm
lide warmer
^to / Eppendorf Pipette
Waterbath (IMV)
aminar Air Flow Unit
CCTV
MV make Photometer
PH Meter
Auto filling-sealing machine (Mini)
Straw Printing Machine
et Printer
Cold Handling Cabinet- Split
Biological freezer
Freezing Racks (Mini)
Distribution Ramp (Mini)
ncubator
Autoclave
Hot Air Oven
Double glass distillation plant
Mlipore water purifying equipment
ETO Steriliser
Magnetic Stirrer
Electronic Weighing balance
WM bulk FS storage Container
with freezing Grill
Bulk FS Storage Container
Geyser
Vacuum Cleaner
Voltage Stabiliser (Central)
FS storage Cont (12000 Mini st)
LN Storage Cont. (50 Lt. Cap)
PC (to connect with Microscope),
Printer & UPS
Fumigator/Humidifier
Thawing Unit (IMV)
LN Pump/Transfer device (IMV)
SS Forceps -18"
Refrigerator
Minimum Standards
< 5 lakh
5 to 10
lakh
10 to 20
lakh
> 25 lakh
1
Nil
1
3
1
1
Nil
1
1
1
1
Nil
1
1
30
1
1
Nil
1
3
2
2
1
2
2
2
2
Nil
1
1
1
2
1
2
1
1
2
1
60
1
2
2
2
2
1
1
1
1
2
1
1
5
2
2
1
2
1
2
Nil
2
2
2
80
1
3
2
2
2
1
1
1
1
2
1
1
6
2
3
1
2
2
2
Nil
2
4
2
100
1
3
2
4
2
1
1
2
1
2
1
2
1
2
2
2
1
2
3
2
1
2
5
2
1
As per
req.
5
10
1
As per
req.
10
15
1
As per
req.
12
20
1
As per
req.
15
20
2
1
2
2
6
1
1
2
2
2
2
2
6
2
2
2
2
6
2
27
6
2
(B). Miscellaneous Equipment
Items
Sr.
No.
1 AV Stand
2
Test Tube Stand
3
Auto Pipette Stand
4
Plastic Tips Holder
5
Spirit Lamp
6
Timer/ stop watch
7
Scissors Straight- 6°
8
Hot Air Blower/ Hair Dryer
9
Trolley for Movement of LNC
10 Anaesthetic Trolley
11 PC with Printer + Software
(SSMS)
12 Glass wares
13 Air-conditioner with Hepa filter
< 5 lakh
5
5
2
2
2
2
2
1
5
1
1
5 to 10
lakh
5
5
3
3
2
2
4
2
5
1
1
10 to 20
lakh
5
5
5
5
2
2
4
2
10
1
2
>25
lakh
5
5
5
5
2
2
4
2
10
2
2
In sufficient quantity
1 ton for 100cft area with remote
(C). Semen despatch section
1
2
3
4
Phase Contrast Microscope with
biotherm
IMV Waterbath
Thermocol box
Long Forceps
1
1
1
1
1
2
1
2
8
1
2
8
1
3
8
8
(D). Quality Control Laboratory
Phase Contrast Microscope
IMV Water bath
Laminar Airflow Unit
Autoclave
Hot Air Oven
Incubator with digital Temp
Haemocytometer
Refrigerator
Gas connection
Water bath
Glass ware/Media/ chemicals
Table.Chair, Almirah
Misc. Items
Minimum Standards
28
1
2
3
4
5
6
7
8
9
10
11
12
13
1
1
1
1
1
1
2
1
1
1
Adequate
11
11
Annex - X
Qualifications recommended for Semen Station personnel
1. Officer-in-charge
MVSc. in Animal Reproduction / Livestock Production & Management, trained in
semen production.
2. Senior Veterinary Officer
MVSc. in Animal Reproduction / Livestock Production & Management, trained in
semen production.
3. Veterinary Officer
MVSc. in Animal Reproduction / Livestock Production, trained in semen production/
Livestock management.
4. Quality control Officer
MVSc. in Animal Reproduction / Vety. Microbiology, trained in carrying out all tests to
determine quality of semen samples as per MSP.
5. Lab Technician
Bachelor degree in Science / Microbiology / Bio-chemistry, in semen production and
quality control. Persons having post-graduation degree are preferable.
6. Semen Collector
Diploma in livestock management or stockman training course with two years
experience in livestock farm / semen laboratory.
7. Lab Attendant/ Bull Handler
High school / 8th standard, should be able to read and write and with two years
working experience in semen station/ bull station/livestock farm.
For semen & LN Distribution
1. Stockman (Semen & LN Distributor)
Stockman with two years experience, trained in transfer of straws & distribution
2. Stockman (Semen Despatch}
Stockman with two years experience, trained in examination of semen under
microscope.
Minimum Standards
29
Annex - XI
Job responsibility of the
The QCO shall be responsible for the quality control work pertaining to semen
production and any other related work as may be required from time to time. He shall
be responsible to carryout the following:
a). Bacteriological tests:
Neat semen
Frozen semen
Lab environment
AV washing
-
Randomly two samples daily
Randomly eight samples daily, all bulls to be
covered once in a quarter
Plates shall be placed daily at different
location in the processing room, AV room,
washing room, pass box, Incubator, Laminar
airflow unit ,etc. to check bacterial load
Randomly two AVs once a week.
b). Morphological abnormalities :
For adult bulls as and when there is report
about poor semen quality, particularly of
crossbred bulls
- Young bulls four to six times before putting
for regular collection. Thereafter, based on
the report of poor quality of semen
Depending on the report of poor quality of semen or certain abnormalities, such
bulls shall be kept under observation and semen samples examined twice a week
for 4 to 6 weeks. Semen from such bulls shall not be frozen until cleared by the
QCO.
Morphological abnormalities c). Acrosome integrity test:
Frozen semen samples of all adult & young
bulls in each quarter
d). Validation of photometer
Sperm concentration of neat semen shall be
measured by Haemocytometer separately for
cattle & buffalo bulls for validation of IMV
photometer (as per MSP) - twice a year
e). Sperm concentration of frozen semen sample :
- Randomly two samples per week each of
cattle and buffalo.
Minimum Standards
30
f). pH measurement:
- washing of sterilized glassware / rubberware
shall be checked for pH once a week
g). HOST
-
everyday four samples after 24 hrs. PTM
h). Incubation Test
-
every day 4 to 6 samples
j). Percent intact acrosome
-
all bulls to be covered once a quarter
Semen samples frozen routinely shall not be checked by those who are involved for
semen processing. Semen samples shall be checked by the QCO for PTM after 24
hours of freezing. The QCO will be the final authority to accept or reject semen
samples. In addition, he will be testing the quality of straws or any other consumable
materials used in semen station.
NB. Depending on the work load, the QCO should be assisted by a Lab technician
and Lab attendant.
arb:C/alldoc/Min Stn/Min. Sid. for Govt.doc (April1 2005)
Minimum Standards
31
N.D.D.B.
SEMEN BANK
DRAWING NO.
ANAND 388001
TOP PLAN
LAB 01.A4
SEMEN BANK-LAB.01A4
2 8 -1 1 - 0 2
N.D.D.B.
ANAND 388001
BULL SHED GROUND
FLOOR PLAN - SECTION
DRAWING NO.
AR-02-05.A4
BULL SHED-05.A4
- S3 -
28-11-02
'i^jii*^
^'.'^'.^^-»>,-,