Pre-Feasibility Study: Livestock Semen Production Unit

Pre-Feasibility Study
LIVESTOCK SEMEN PRODUCTION UNIT

(Cattle & Buffalo)
Small and Medium Enterprise Development Authority

Government of Pakistan
www.smeda.org.pk
HEAD OFFICE 6t Floor, LDA Plaza, Egerton Road, Lahore Tel: (042) 111-111-456, Fax: (042) 6304926-7 helpdesk@smeda.org.pk
REGIONAL OFFICE PUNJAB 8th Floor, LDA Plaza, Egerton Road, Lahore. Tel: (042) 111-111-456 Fax: (042) 6370474 helpdesk@smeda.org.pk REGIONAL OFFICE SINDH 5TH Floor, Bahria Complex II, M.T. Khan Road, Karachi. Tel: (021) 111-111-456 Fax: (021) 5610572 helpdesk-khi@smeda.org.pk REGIONAL OFFICE NWFP Ground Floor State Life Building The Mall, Peshawar. Tel: (091) 9213046-47 Fax: (091) 286908 helpdesk-pew@smeda.org.pk REGIONAL OFFICE BALOCHISTAN Bungalow No. 15-A Chaman Housing Scheme Airport Road, Quetta. Tel: (081) 831623, 831702 Fax: (081) 831922 helpdesk-qta@smeda.org.pk
March, 2008
Livestock Semen Production Unit (Cattle & Buffalo)
DISCLAIMER
The purpose and scope of this information memorandum is to introduce the subject matter and provide a general idea and information on the said area. All the material included in this document is based on data/information gathered from various sources and is based on certain assumptions. Although, due care and diligence has been taken to compile this document, the contained information may vary due to any change in any of the concerned factors, and the actual results may differ substantially from the presented information. SMEDA does not assume any liability for any financial or other loss resulting from this memorandum in consequence of undertaking this activity. Therefore, the content of this memorandum should not be relied upon for making any decision, investment or otherwise. The prospective user of this memorandum is encouraged to carry out his/her own due diligence and gather any information he/she considers necessary for making an informed decision. The content of the information memorandum does not bind SMEDA in any legal or other form.
DOCUMENT CONTROL
Document No. Revision Prepared by Approved by Revision Date Issued by PREF-51 2 SMEDA-Punjab GM Punjab March, 2008 Library Officer
PREF 51/March, 2008/2
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1. 1
INTRODUCTION
Background1
Livestock is an important component of Pakistans population since 30 - 35 million rural population is involved in livestock raising. Average household holdings are 2-3 cattle/ buffalo, 3-4 sheep/goats and 10-12 poultry per family which contribute 35 to 40 percent of their income. Government gives high priority to its development and is focused on private sector led development of livestock. Underpinning the importance of livestock the government instead of maintaining the previous practices of making livestock a part of the National Agriculture Policy has formulated an independent Livestock Development Policy, providing a framework for accelerated development of livestock. This policy not only addresses the need of the small livestock farmers for whom livestock is a supplementary income source but also includes measures to develop small and medium livestock enterprises and an incentive framework for setting up large livestock farms. The Livestock Census 2006 carried out by the Agricultural Census Organization have been released. According to the Census, the share of livestock in agriculture growth has jumped from 25.3 percent in 1996 to 49.6 percent in 2006. The higher growth in the livestock sector was mainly attributed to growth not only in the headcount of livestock, which is commercially important but also in the milk production. The details of the Census results are documented in Table 1-1. Table 1-1 Comparative Status of Livestock Population between 1986-96 & 1996-20062
1. 2
Project Brief
The proposed project is for establishing a semen production unit (SPU), where the adult bulls of good genetic background are maintained. Semen from the bulls is collected twice a week. After laboratory evaluation, semen is diluted and packed. The deep-freezing of the semen in liquid nitrogen is carried out for the preservation of spermatozoa for longer periods. This semen is then utilized for artificially inseminating the female animals for improving the breed of the offspring, ultimately leading to better milk and meat yields.
1 2
Pakistan Economic Survey 2006-07 Pakistan Economic Survey 2006-07
1. 3
Rationale and Justification of the Investment Opportunity
Reproductive efficiency is the most economically important trait in farm animals. The profit obtained from milk and meat is directly related to the fertility of the respective farm animal. There are two ways of breeding of farm animals, naturally and by artificial insemination. Presently, there are very few large farmers, who raise the breeding bulls specifically for their own herds. Most of them do not have access to any breeding bull. Artificial insemination (AI) program started during 1972-73 in the country has now gained much acceptance and popularity among livestock breeders and farmers with increasing demand for more AI services. Unfortunately, (AI) services of livestock and dairy development department hardly cover 5% livestock population of the Punjab. Punjab Livestock Census 2000 indicated hundreds of villages without proper AI service and breeding bull. The current status of cows & buffaloes artificially inseminated is presented in Table 1-2. The figures are based on complete census of livestock for all the provinces of Pakistan. The ratio of a breeding bull to available breed-able buffaloes in Punjab has been found to be 1:66. Households reporting and number of cows and buffaloes artificially Table 1-2 inseminated (2006)3
The acute shortage of buffalo breeding bulls has been identified in six districts of Punjab where the ratio ranged from 1:205 to 1:411. Table 1-3 Bull: Buffalo ratio in 6 districts of Punjab Breeding Bull Ratio to Breed-able Buffalo 1:411 1:297 1:256 1:232 1:216 1:205 District Gujrat Jhelum Attock Chakwal Rawalpindi Khushab
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Agriculture Census Organization, Statistics Division, Govt. of Pakistan.
In view of the prevailing shortage of breeding bulls and 7.14% of the AI coverage for Buffaloes & 11.10% AI coverage for cows, there is an immediate need to establish semen production units (SPU), for increase in buffalo production. These SPU will produce and supply semen to meet the demand of farmers & commercial entrepreneurs in the country. Quality semen is a prerequisite for breed improvement and to develop a viable and functional artificial insemination program. 1. 3 Proposed Capacity
Initially, a minimum of 10 bulls has been suggested for establishing SPU. Cow bulls have also been proposed in the SPU beside buffalo bulls. A mix of six buffalo bulls, two Friesian and two Sahiwal/Cholistani bull depending upon the location of the SPU are recommended. 1. 4 Project Cost
The cost of proposed project is approximately Rs 2.632 million.
CURRENT INDUSTRY STRUCTURE
Most of the SPUs are being operated in the public sector, which are catering to the needs of 5% livestock population of the country through its own network of artificial insemination centers. 2. 1 Demand Qadirabad, Distt. Sahiwal Karianiwala, Distt. Bahawalpur Kharimurat, Distt. Attock Kalourkot, Distt. Bakkhar
There are four SPUs in the country at the following locations.
These SPUs are producing 2.2 million semen doses annually, whereas, the population of breed able buffaloes and cows in Pakistan is over 30 million, which needs over 43 million doses annually. In the absence of desired number of breeding bulls, there is high demand of semen doses in the country. 2. 2 Total Market Size and Growth
Currently, public sector is meeting the demand of 5 percent of the breed-able buffaloes and cows in the province. The Nili-Ravi buffalo breed is the most superior dairy breed in the world. There is always a demand for good quality buffalo semen of Nili-Ravi breed in the country. The Sahiwal cattles are the best dairy cattle, suited for the tropical regions of the world; its high milk yield and resistance against ticks are the characteristics, which contribute to the high demand of these animals. Besides meeting the domestic market demands of frozen semen, there is a potential for exports as well.
MARKETING
Marketing is the critical component for this projects success. The milk processing plants are in the process of establishing networks of artificial insemination services for the
genetic improvement of Livestock. The proposed SPU will cater to the needs of breed-able population of cattle and buffaloes through these networks. There is an acute shortage of good quality frozen semen, both for buffalo and cattle breeds. For the marketing of semen, following groups will be approached: Large Livestock Farms. Artificial Insemination Technicians. Public Sector Breed Improvement Department. Importers.
ANIMAL REQUIREMENT
The proposed number and details of the animals required for the project are given in the table: Table 4-1 Animal Requirement Description Buffalo Bulls Friesian Sahiwal/Cholistani Total No of Animals 6 2 2 10
The bulls would be used for reproduction activity for 10 years, and after that, they would be replaced with younger bulls. Young adult breeding bulls of known pedigree will be purchased from livestock experiment stations of the respective breeds or from livestock breeders listed in the progeny-testing program of Livestock and Dairy Development Department. A breeding soundness examination (BSE) of dairy bulls, to be selected and raised at SPU, will be required just before their purchase. For dairy bulls to be raised at SPU, it is an absolute requirement that bull is born to elite cow inseminated with the semen of progeny tested bull.
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5. 1
TECHNICAL ASPECT
Examination of External Reproductive Organs
5.1.1 Scrotum The scrotal shape of the bull controls testicular temperature. optimum temperature is a few degrees below body temperature. This thermoregulatory function is essential for normal spermatogenesis. Normal bull scrotum should be pendulous, symmetrical and should have a distinct neck at point of attachment with the abdominal wall. Any deviation observed on inspection in size, shape or relative position should be reviewed with suspicion. All physiological (rounded, rotated, cleavage) and pathological (small hypoplastic, asymmetrical, straight sided, high pointed) variations must be observed.
5.1.2 Testes On each clinical examination, testes should be carefully examined and palpated entirely for their size, consistency and free mobility within scrotum and through the inguinal canal. In general, both normal testes are symmetrical in size, uniform in consistency, and are freely moveable up and down. Biometry of each testicle (length, width and thickness) is determined by holding the other pushed dorsally. While doing this, the consistency of its texture is also checked with gentle pressure from thumb and fingers. Resonance and tonicity are the signs of normally developing tissues. Soft, firm and hard textures are indications of some kind of abnormality. Any defect observed (orchitis, asymmetry, excessive fat around testes, hernia, cryptorchidism, hypoplasia, etc.) need to be carefully diagnosed. 5.1.3 Epididymes The epididymes on the side of each testis consists of three parts; caput (head), corpus (body) and cauda (tail). Like the testis, it needs to be palpated to its entire length to record its size, form and consistency. Though the defects of epididymes are rare, yet it must always be scrutinized for segmental aplasia (part of the duct missing), epididymitis, adhesions, tumors, abscesses and spermatic granulomas. 5.1.4 Reproductive Corpulatory Organs This will include examination of sheath, prepuce and the penis. The young bulls should be examined for normal development of the penis and freedom from prepural adhesions and developmental anomalies such as persistent frenulum. The best time to examine the penis is at the time of semen collection. The perpetual opening and sheath should be thoroughly palpated to record any abnormalities present. The veterinarian should always watch for the occurrence of persistent penile frenulum, hair ring around penis, fibropapilomas of penis, eversion of prepuce, penile hematomas, phimosis, paraphimosis and fibrotic preputial orifice. 5.1.5 Examination of Vision Good sight is important to the breeding bulls. The eyes for common problems, like pink eye, scars and squamous cell carcinoma need to be checked. Scars resulting from accidents and infections like keratitis should be treated promptly and properly. 5.1.6 Examination of Teeth The bull needs healthy, strong enough teeth to chew coarse fodder and concentrates. Overgrowth of molars and broken teeth must be diagnosed early through oral examination otherwise he shall loose weight rapidly and this may impair his reproductive performance. 5. 2 Examination of Skeletal Development and Confirmation
5.2.1 Hind Legs Structure Sound rear legs are vital to the breeding capacity of a bull. Pain or deformity in rear legs will not allow bull to readily mount the cow/teaser properly and give good thrust. The bulls
therefore should be evaluated for mobility, interdigital fibroma (corn) and foot abscess should be checked. Since the structural defects of the legs are heritable, and become more apparent and interfere more and more with the bulls breeding ability as the bull ages, such bulls should be culled right away. Special attention should be paid to hoof trimming in stall fed bulls. Similarly, the conformation of fore legs may also be checked. Some bulls show outward direction of feet, which on walking swiftly may stumble by striking with opposite foot. 5. 3 Conformation of buffalo, Sahiwal and F1 cross-bred Bulls
The conformation of the bull is evaluated by checking the breed characteristics of the bull. Buffalo bulls should have Nili Ravi characteristics, black color with star at forehead white four feet and switch of tail, (Panj Kalian). Although such breed charms are not correlated with breeding soundness, yet buffalo breeders give a very high sentimental value to these phenotypic characteristics. Sahiwal bulls should be of dark brown color with black extremities. F1 crossbred bulls of different colors are found. Generally, the farmers prefer black or black and white spotted bull with good height. 5.3.1 Secondary Sex Character The developments of head and neck and hump muscles give a bull-like appearance to male animal. This masculine development takes place in bull as he approaches puberty and is said to be due to normal secretion of androgen hormone. A bull with well-developed body, heavy and deep at forequarters is generally liked. 5.3.2 Evaluation of Sexual Behavior The sexual behavior of bull for convenience of understanding can be divided into; libido (sexual desire) and mating behavior (ability to complete the service). a) Libido Index Libido of a bull is defined as the willingness and eagerness to mount the teaser/cow and attempt service. The libido is graded from 0 to 3, depending upon its performance; 0-for being shy i.e. no desire to move towards cow/teaser. 1- for being Dull, very reluctant to reach the teaser. 2- for being Active, willingly moves towards the teaser. 3-being Aggressive, moves towards teaser in an uncontrolled manner.
b) Mating Behavior Index (MBI) After the bull has reached the teaser/cow, it mounts and hold the teaser/cow to complete the act of copulation or semen ejaculation. During this period, the veterinarian gets an opportunity to record and examine the sequence of events, holding erection of penis, seeking movements (for vagina or artificial vagina) of penis, thrust, ejaculation and dismounting. Any painful condition in the feet, hind legs, back, penis, sheath, or in any part of the internal cavities (transmatic gastritis, peritonitis, nephritis, pleurisy, pneumonia,
etc.) shall inhibit the desire of mating. In these situations, either bull will not mount the teaser at all or may step down after mounting without completing the act. In bulls, it is easy to grade bulls for their mating behavior on the basis of number of attempts taken to mount the teaser for completion of a successful ejaculate. The bull is ranked from 0-3; 0-Poor when the bull ejaculate after more than 4 attempts 1-Fair, when he takes 3-4 attempts 2-Good, if it takes between 2-3 attempts 3-Excellent, when the bull ejaculates in less than 2 attempts. The bulls falling in categories 0 and 1 for libido or mating behavior or both shall stand disqualified. 5.3.3 Evaluation of Semen Production Ability It is now recognized fact that scrotal circumference of a bull is an accurate predictor of his sperm production ability (particularly in young bulls) and gives an accurate indication of future testicular function. In buffalo bulls, scrotal circumference has been found to be correlated with age, body weight, and sperm 0production. Similar correlation exists in cow bulls. 5. 4 Management of bulls4
5.4.1. Housing Housing may be closed, semi-open or open. Bulls in tropical and subtropical conditions require protection from heat and adequate ventilation. Shade trees, shade cloth and thatch are effective. Fine water sprays with fans can be used to cool Bos taurus bulls under hot conditions. Bulls should be housed securely so there is no chance of escape and interaction with other bulls, staff and the general public. 5.4.2. Feeding A balanced ration should be fed. This could be home grown or bought in or both. Care should be taken not to over-feed bulls as fat deposition in the inguinal canal negatively affects fertility. Condition score is an important guide to nutritional requirements. Breeding bulls should have a score of 3 on a scale of 1-5. Bulls should have access to mineral licks and clean water ad libitum. 5.4.3. Handling The establishment of a firm relationship between the handler and the bull is essential and can not be overemphasized. The bull should be at ease when he is handled and the handler should not feel threatened. The proper application of a bull nose ring is required as soon as the bull arrives at the centre. The bull should be handled by both a halter and the bull ring. Care should be taken to use the ring only when the bull becomes unruly and difficult to handle by the halter alone. Bulls should be led by the halter and not by their nose (Fig. 1). Bulls should always be handled in such a manner that semen production is optimised. This includes taking note of all aspects of the physiology of male sexual behaviour. Negative stimuli should be avoided in the collection area. This includes pain delivered via the nose
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TECDOC-1480, IAEA International Atomic Energy Agency
ring, which could lead to low libido (e.g. many of the difficult, slow, low libido Bos indicus bulls may have been made that way by poor training and handling techniques).
Fig. 1. Young bulls should be taught to lead with the halter. The nose ring is used sparingly for restraint and not at all when the bull is immediately behind the teaser. 1.4.4. Health Once the bulls have passed all the quarantine tests for disease control, normal routine preventive medicine should be practiced in the AI Centre. Care should be taken that bulls remain in excellent health for continuous semen production. The necessary vaccinations, regular deworming and control of ectoparasites should be implemented to meet national and regional requirements. Adequate exercise and regular hoof care should be provided. Continuous monitoring of diseases should be undertaken whether statutory or not. All quarantine requirements, including restrictions in the movement of animals and personnel, must be strictly observed. It is in the best interest of the AI Centre to be able to certify at all times that all animals are fit to produce semen for sale and distribution. 1.4.5. Records A complete history of every animal should be kept from the time of arrival until the day of departure from the centre. All incidents, ailments and medications should be recorded. 5. 5 Semen Technology and Field Practices
5.5.1. Semen technology a. Collection area and facilities The semen collection area should be as close as possible to the semen evaluation laboratory (not more than 30 m). For teaser bull restraint a stanchion made from strong metal bars or smooth treated wooden poles and timber is recommended (Fig. 2). The floor
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of the collection site should not be slippery. It can be made of rough concrete or a dug-out filled with sand and sprinkled with water to avoid dust. Rubber mats can also be used.
Fig. 2. Strong timber construction of stanchion for teaser bull restraint. Note the non-slip floor. Facilities for the restraint of bulls awaiting their turn for semen collection should be near enough to enable them to see clearly the mounting bull and serving area. The collection area should be ringed with strong metal bars or timber for the safety of people and the bulls themselves. The construction should be high enough to protect the full height of an average person (1.75 m). Spaces between rails should be small enough to prevent a bull getting his head through. Escape spaces in the surrounding fences should be placed at regular intervals. The collection area should be sheltered and must have adequate ventilation and light. b. Preparation of bulls The semen donor bulls must be housed under clean dry conditions and should be washed and cleaned before they arrive at the collection area. The washing area should not be more than 20 m from the serving area and should be made of rough concrete with a slanting floor to facilitate drainage of water, dung and urine. Adequate clean water with reasonable pressure should be provided through a hose pipe at this area. Prior to cleaning, the perpetual hair should be cut short, leaving a tuft of 2 cm length all round. Ordinary washing soap and a soft brush should be used to clean the bulls. During cleaning, emphasis should be put on the lower abdomen and the perpetual area. If necessary, washing of the perpetual sheath with normal saline solution can be done once every week or fortnight to reduce bacterial contamination of semen. Disinfectants should not be used. Clean, dry paper towels should be used after washing to remove excess water.
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If the teaser bull or steer is dirty, its back should be cleaned with water and soap and dried thoroughly. An apron may be used if necessary. There is little risk of contamination of the penis or the semen if the teaser is clean and collection technique is good, allowing no or little contact of the penis with the teaser. c. Preparation and sterilization of equipment and materials All equipment used for the collection, evaluation and processing of semen must be clean and sterilized. The following procedures are recommended i) Glassware Wash with detergent containing 2% Na2Co3. If glassware has become cloudy through repeated use, leave it submerged in potassium dichromate solution (K2Cr2O7, 8 g; H2SO4, 12 ml; distilled water, 100 ml) for 24 hr. Wash with tap water Rinse with distilled water Dry and cover in clean paper or aluminum foil Place in hot air oven (160C) for 30 minutes Transfer to a closed, dust free incubator (37C) ii) Rubber materials Wash in detergent Wash repeatedly in tap water Boil for 10-15 minutes Swab dry Store in a dust-free chamber fitted with ultraviolet radiation. iii) Buffers After preparation, autoclave at 120C and 15 lb pressure for 20 minutes Fresh eggs of Grade A should be collected from pathogen free flocks and the shell washed and swabbed with 70% alcohol d. Artificial vaginas An outer rubber barrel (usually 45cm long) with rough inner rubber liner that is nonspermiotoxic is recommended. The inner liner should periodically be checked for possible leakage. The rubber cones should be also non-spermiotoxic and a correctly labeled collection tube should be attached. A jacket should be provided for the cone to prevent breakage and avoid direct exposure to sunlight. Rubber bands for holding on the cones and the two ends of the reflected inner lining onto the outer barrel should be strong. A lubricant that is sterile, non-spermiotoxic, non irritant to the penis and easily washable (eg. KY jelly or white Vaseline that has been sterilized by boiling) should be applied sparingly and just before collection at the entrance of the artificial vagina (AV, Fig. 3). The lubricant can be replaced by a small amount of diluent to moisten the entrance to the artificial vagina. Water for the outer jacket filling should be warmed to 60oC. Enough of this should be poured into the inner chamber to provide the required pressure. This quantity may range from 500-750 ml. Inner temperature after lubrication should range between 40-45oC. Assembled AVs should be kept in incubators at 55-60oC. If there is a delay between preparation of the AV and collection, the temperature should be checked. Just before collection, excess water is poured off from the AV and enough air blown in to provide adequate internal pressure.
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Fig. 3. Preparation of the artificial vagina; lubricating with a glass rod using KY jelly. e. Electro Ejaculators Electro ejaculators should only be used when absolutely necessary. Only lame or injured bulls should be subjected to the technique. Good training and good handling procedures allow most bulls to be collected with the artificial vagina. The prepuce should be washed and dried. The rectum should be emptied of faces and the probe inserted to lie over the seminal vesicles and ampullae. Stimuli should be applied with great care to achieve a very slow and gradual increase in intensity. f. The collector A collector should be selected on the basis of his/her ability, enthusiasm and experience to work with livestock. Protective gear should include gum boots with steel or wooden-toed caps, apron, head cap and thin half length plastic hand gloves. g. Collection procedure It is advised to collect the semen early in the morning. Bulls should be led, preferably using a halter, to the teaser in a gentle friendly manner by the handler, who should pay attention to the temperament of the particular bull. The bull should be allowed to watch other bulls mounting before collection. He is led around behind the teaser and may be allowed to mount other bulls. Two false mounts are usually given. These measures promote good sexual excitement, which improves the quality of semen by cleansing the urethral passage and increasing the amount of semen collected. The bull is then allowed to mount for the first collection. At this time the collector shall gently grasp the prepuce behind its opening and direct the fully erected penis into the lubricated end of the AV. The penis should not be touched. The handler may rest his shoulder against the bulls flank and move with the movement of the bull as he thrusts. The AV should be held so that the bull withdraws as he dismounts, and
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should not be pulled away from the penis. The ejaculate should be taken immediately to the evaluation room. Handling of semen should be always done with great care to avoid cold shock, contamination, excessive agitation and direct sunlight. h. Evaluation of semen i) Macroscopic examination The semen should be transferred to a water bath maintained at 351oC. Visual evaluation for volume, colour, consistency/density, odor and observation for presence of foreign material (blood, pus cells, dung, hair, etc.) shall be made and recorded. If dung or hair is found in the semen, filtration with a special semen filter is advised. b) Microscopic examination Microscopic evaluation is done using a simple or phase contrast microscope for mass activity (wave motion) and individual motility. Determination of concentration is done with a hemocytometer or a calibrated photometer. At this point, if required, smears can be made for morphological studies and live/dead count. Nigrosin-eosin stain is recommended. Buffered nigrosin-eosin solution is mixed with a drop of semen and smeared on a glass slide for morphological examination. It should be dried and examined under oil immersion. Automated computerised machines for recording motility and concentration and calculating the required extensions are now frequently used in AI centres that can afford them. Semen used for artificial insemination should be of high quality. The following are guides to the values of semen characteristics in the bull that indicate good reproductive function: Motility (moving actively forward): > 60% Concentration: > 500 million /ml Live sperm: > 70% Abnormal sperm: < 20% (range for bulls with good fertility is 812%) Proximal droplets: < 4 %; Distal droplets: < 4% Tailless: < 15%; Singly bent tails: < 8%; Double bent tails: < 4%; Coiled tails: < 3% Cells other than spermatozoa: none, or very few leucocytes or epithelial cells. A further technique that can be used to evaluate the semen is the hypo-osmotic swelling test (HOST), which indicates the functional integrity of sperm membranes. Centers should develop a system of morphological assessment and guidelines for limits beyond which semen is discarded. Morphological examination is generally reserved for borderline samples. The assessment is also valuable in helping to reach a diagnosis when a bull begins to fail to produce semen of processable quality as assessed by concentration and motility. The definition of motility is often ambiguous. Since the important criterion is progressive forward motility, this should be the basis for judgment. If there are 70% or more of spermatozoa moving actively forward the semen sample is of good quality and acceptable for processing. If there are 40% or more of spermatozoa moving actively forward after freezing and thawing the quality is acceptable for AI. For election/rejection purposes it does not matter very much if the others are slow, swimming backwards in circles (singly bent tails) or immotile. However, these characteristics are important for diagnostic purposes, because they help to define the disturbance of function.
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Some systems of evaluation characterize motility as follows: (a) % direction motility (moving forward); (b) % local motility (wiggling around without going forward); and (c) % no movement (possibly all dead). To judge this under the microscope, the general picture is first assessed, and then the type of motility of those moving is assessed. To be acceptable, more than 50% should be moving, and of these more than 70% should show progressive motility. Many artificial breeding centres have standardised their own way of assessing semen quality and, provided they serve the purpose effectively, can be recommended for use. i. Extension The Extender type and extension ratio depends on the type of semen produced: deep frozen semen (DFS), chilled semen (CS) or room temperature semen (RTS). For DFS the recommended diluents are: Egg yolk - citrate - glycerol extenders Skimmed milk - egg yolk - extenders Tris buffer - egg yolk - glycerol extenders For CS and RTS the recommended extenders are: Coconut milk - egg yolk extender Egg yolk - citrate extenders (cattle) Tris - egg yolk extenders (buffalo). j. Processing and packaging A standardized daily routine should be adopted for all types of semen processing. For example, the following routine is recommended: Extender preparation; Semen collection and evaluation; Extender A at 35oC added to semen 1:1 and allowed to cool to room temperature (approximately 20 minutes); Complete dilution with Extender A at room temperature and placed in 45oC for at least 4 hours; Extender B is held at 45oC and added in two steps, 30% and then 70% at that temperature; Fill, seal and label straws at 45oC; Place straws on freezing racks in liquid nitrogen vapor to -140C over 810 minutes (straws should be 5 cm above the liquid nitrogen surface; in the absence of freezing machines this step can be done in a large semen storage tank or a big polystyrene container containing liquid nitrogen); Place racks in liquid nitrogen at -196oC; Collect straws with a gloved hand and store in goblets in liquid nitrogen; Wash and sterilise glassware for the next day. In this system the extender is added in two fractions. Fraction A contains no glycerol, fraction B contains 14% glycerol. The final concentration of glycerol is 7%. The 4 hour time lapse between adding fraction A and the first part of fraction B is to allow antibiotics to work before they are inhibited by glycerol.
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The common types of packaging used for processed semen are: DFS - packaged and sealed in straws, mini (0.25 ml) or medium (0.5 ml), or as pellets. Straws and pellets contain a minimum of 2030 million spermatozoa per dose. RTS and CS - packaged and sealed in ampoules or airtight vials of 1.0 ml, containing 1520 million spermatozoa. In some cases this can be reduced to 5 million spermatozoa (e.g. the caprogen diluted semen used in New Zealand). Vials containing chilled semen are transferred to a beaker with water at 35oC and the beaker is transferred to the refrigerator at 46oC. k. Preservation and storage DFS is preserved in liquid nitrogen at 196oC. Transferring of semen must be done quickly. Canisters containing packages when raised from the tank should remain in the neck of the tank for less than 10 seconds. Liquid nitrogen is dangerous and must be handled carefully. CS is refrigerated at 45oC. RTS is held at ambient temperature (1826oC). The containers of straws, ampoules and pellets should be properly labeled and records maintained on their location and contents. l. Post packaging quality control Motility of samples from processed batches of semen should be checked before dispatch. Post thaw motility should be 40% or more for DFS. All semen storage containers should be regularly checked for liquid nitrogen level and replenished as required.
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MACHINERY REQUIREMENT
Detail of Equipment Requirement Qty 4 1000 300 2 1 1 150 10 1 1 1 1 6 1 1 1 1 1 1 1 1 1 1 1 3 1 1 1 Unit Kanal Sq. Ft Sq. Ft Nos. No. No Sq. Ft Nos. Nos. Nos. Nos. Nos. Nos. Nos. Nos. Nos. No. No Nos. Nos. Nos. Nos. Nos. Nos. Nos. Nos. Nos. Nos. Cost per Unit 62,500 400 600 20,000 25,000 10,000 250 50,000 20,000 50,000 60,000 65,000 5,000 75,000 10,000 8,000 50,000 150,000 15,000 10,000 10,000 25,000 15,000 25,000 75,000 30,000 11,000 50,000 Total Cost (Rs.) 250,000 400,000 180,000 40,000 25,000 10,000 37,500 500,000 20,000 50,000 60,000 90,000 30,000 75,000 10,000 8,000 50,000 150,000 15,000 10,000 10,000 25,000 15,000 25,000 225,000 30,000 11,000 50,000 2,401,500
The detail of the equipment required for running the project is given in below table: Table 6-1 Description Land Bull Shed Laboratory (3 rooms of 10'x10' each) Office Furniture for Staff Computer Printer Feed Store Cost of Bulls Refrigerator Microscope Incubator (50 Litre) Water Distiller (4L / hr) Artificial Vagina Sterilizer Weighing Balance Magnetic Stirrer Filling & Sealing Machine Cold Cabinet Cattle Crush with Installation Fodder Chopper Cart Bull for Cart Water Pump Air Conditioner Liquid Nitrogen Container (50 Liter) Liquid Nitrogen Container (10 Liter) Liquid Nitrogen Container (2 Liter) Generator Total
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HUMAN RESOURCE REQUIREMENT
A bull attendant, veterinary technician and veterinarian will be required as a permanent staff, whereas, temporary labor will hired as and when required. The detail of the human resource requirement for the proposed project is given in table: Table 7-1 Detail of Human Resource Requirement No 1 1 1 2 1 1 2 12 Unit Monthly Salary Annual Salary Nos. 30,000 360,000 Nos. 25,000 300,000 Nos. 10,000 120,000 Nos. 5,000 120,000 Nos. 10,000 120,000 No. 6,000 72,000 Nos. 4,000 96,000 Months 10,000 120,000 1,308,000 Description Manager SPU Pay of Veterinarian Pay of Technician Bull Attendant Accountant Security Guard / Gate Keeper Sweeper Traveling/Promotion Expense/Office Expense Total
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8. 1
AREA REQUIREMENT
Land Requirements
About 0.5 acres (4 kanal) of land will be purchased for a SPU of 10 adult bulls and in addition to that 1 acre of land will be acquired on rent. The land will be used for building a shed for bulls, feed store, bull exerciser, cattle crush (for semen collection), laboratory and open area for animals. Table 8-1 Description Bull Shed Laboratory Feed Store Total 8. 2 Detail of Covered Area Requirement Area in sq. ft 1000 300 150 1450 Construction Rate per sq. ft 400 600 250 Total Cost in (Rs) 400,000 180,000 37,500 617,500
Recommended Mode for Acquiring Land
Land on rental is a better option for a new investor. Land on rental is available in rural and suburban areas. The higher livestock population regions of the country will be suitable for establishing a SPU. Rental payment will be paid in advance on yearly basis. This particular land will be used for growing fodder for the animals. Table 8-2 Land Rental Detail Rental per Acre 20,000 Total Amount in (Rs) 20,000 Description 8 kanal of Land
18
8. 3
Bull Housing
An airy shed will be constructed to protect the animals from extremes of climate to reduce stress. Trees will be planted on east & west sides of the open area to provide natural shades. A manger will be made in the shed and the bulls will be tied in head to head position. In the open area, the bull will be offered feed/forage in individual mangers and tied up separately. Chains on their neck or attached to legs will be used to tie all the bulls. The covered area and open area requirements for the breeding bulls are given below: Table 8-3 Floor Space Requirement of Breeding Bulls Floor Space Requirement m2 Covered area Open Paddock 9.0 40.0 Height of Shed (cm) at eves 175 in medium and heavy rainfall areas and 220 in semiarid and arid areas Type of Animals Cattle/Buffalo Bull
The feeding and watering space requirements of breeding bulls are presented in below table: Table 8-4 Type of Animals Feeding & Watering Space Requirements of Breeding Bulls Space/animal (cm) Pen of 10 animals Manger Water trough length, length, (cm) (cm) 600-750 60-75 Manger/Water Trough Width, Depth, Height of (cm) (cm) inner wall 60 40 50
Buffalo/ Cattle Bull 8. 4
60-75
SPU Boundary
A fence can be used as a boundary of SPU. It protects the facility from intruders, and keeps the animals inside. The fence must be strong and durable. Wire fences are practical and longer lasting. Plain-woven wire mesh or barbed wire is used for farm fences. The height of the fence should be 1.5 meters; the recommended gauge of wire is 9 and the space between strands of wire is 35 cm. The top most strands can be of barbed wire. The end and corner posts of fences should be well fixed in a concrete bed and further strengthen by a brace, also fixed in concrete.
9
9. 1
OTHER REQUIREMENTS
Electricity
Electric power is required for forage chopping machine, electric fans, electric bull exerciser, water pump and laboratory instruments. A generator will be used in case of electricity breakdown.
19
9. 2
Forage
The daily green forage requirements will be met by purchase, whereas, dry forage i.e. wheat and rice straw may be stocked during the harvest season. A bull cart will be purchased to haul the green forage from the market/field on daily basis. 9. 3 Concentrate
Supplementing of forage with concentrate is required to improve semen quality/quantity and vigor of the breeding bulls. The concentrate mixture as per requirements, based on seasonal agro-industrial by-products and cereal grains, will be prepared at the SPU on weekly basis. The common ingredients used are maize, wheat middling, wheat bran, rice bran, canola meal, rape seed meal, molasses and mineral mixture.
10 REGULATIONS
1. Semen Production Unit can be started in rural areas. A no-Objection Certificate from Directorate of Breed Improvement of the Livestock Department is required to establish SPU. 2. There is no price regulation governing the sale of frozen semen. However, Livestock and Dairy Development Department has its own price structure for the sale of semen and/or insemination.
11 KEY SUCCESS FACTORS

1. Maintenance of bulls in good condition suitable for semen collection is highly essential for the success of Semen Production Unit. 2. The best condition for breeding is one in which the animal is healthy, receiving sufficient nutrients but at the same time devoid of fattening. 3. The bulls, which receive plenty of exercise, will usually produce larger ejaculates containing more sperm of higher activity. 4. A properly balanced ration should be fed containing the right mix of proteins, minerals and vitamins. Green fodder must be available. In fact, feeding and exercise may be judiciously controlled to obtain a condition of thrift in which the animal is neither fat nor thin and weak. 5. There is no hard and fast rule for the number of services. It varies between individuals and breeds. It is of great importance that males should be used regularly and not too much at one time, and too little at another. Ejaculation every other day is within the capacity of an adult bull. 6. Bulls should be docile, well trained to be led and free from vices. The use of a shield over eyes often assists the control of a bad tempered bull and prolongs its breeding life. Most bulls will serve best in familiar surroundings. A regular routine should be followed and bulls should be handled in the same manner each time. It is sound practice to provide cool quarters to the males with adequate shade and cool drinking water. High environmental temperature is harmful to the reproductive performance of males.
20
7. The animals should be vaccinated against foot and mouth and hemorrhagic septicemia as per schedule and de-wormed against internal parasites on regular basis.
12 THREATS FOR THE BUSINESS

1. The breeding bulls should be free from venereal disease, if they acquire these diseases it will affect the semen production performance of the animals. Moreover, the semen of these bulls will not be fit for use. 2. If the animals are not protected properly against the viral disease there is every threat of loss of animal.
21
13 PROJECT ECONOMICS
13.1 Project Cost/Capital Requirements Project Costs Land purchase (4 Kanals) Building & Infrastructure Machinery & Equipment Bulls Total Capital Cost Prepaid land rent Minimum Cash Balance Raw material Inventory Pre paid Insurance Working Capital Total Project Cost Table 13-1 Equity Debt Total Table 13-2 Project Returns 3,457,957 47% 2.36 NPV IRR Payback Period (Years) Project Financing 50% 50% 1,316,968 1,316,968 2,633,936 250,000 617,500 1,034,000 500,000 2,401,500 20,000 100,000 92,436 20,000 232,436 2,633,936
22
14 FINANCIAL ANALYSIS
14.1 Projected Income Statement
Revenue form Sale Other Income Total Revenue Cost Sales Cost of Goods Sold Medicine, Vaccination Direct electricity, Fuel Charges & Generator Exp. Total Cost of Sales Gross Profit General Administration & Selling Expenses Administration Salaries Traveling/Promotion Expense/Office Expense Land lease rental expense Mortality Expense Insurance Administrative Overheads Depriciation Expense Total Administration & Selling Costs Operating Income Earning Before Interest & Taxes Interest Expense on Long Term Debt Total Interest Cost Earning Before Tax Taxable Income for the Year Tax Net Profit/(Loss) After Tax Balance Brought Forward Total Profit Available for Appropriation Dividend Balance Carried Forward Year 1 4,940,000 4,940,000 1,103,731 5,500 180,000 1,289,231 3,650,769 1,188,000 148,200 20,000 26,250 20,000 49,400 183,025 1,634,875 2,015,894 2,015,894 184,376 184,376 1,831,518 1,831,518 457,880 1,373,639 1,373,639 686,819 686,819 Year 2 5,187,000 5,187,000 1,202,634 6,050 198,000 1,406,684 3,780,316 1,247,400 155,610 22,000 28,875 20,000 51,870 166,329 1,692,084 2,088,232 2,088,232 156,483 156,483 1,931,749 1,931,749 482,937 1,448,812 686,819 2,135,631 1,067,816 1,067,816 Year 3 Year 4 5,446,350 5,718,668 5,446,350 5,718,668 1,310,854 6,655 217,800 1,535,309 3,911,041 1,309,770 163,391 24,200 31,763 20,000 54,464 151,222 1,754,808 2,156,232 2,156,232 124,685 124,685 2,031,548 2,031,548 507,887 1,523,661 1,067,816 2,591,477 1,295,738 1,295,738 1,429,294 7,321 239,580 1,676,195 4,042,473 1,375,259 171,560 26,620 34,939 20,000 57,187 137,549 1,823,113 2,219,360 2,219,360 88,435 88,435 2,130,925 2,130,925 532,731 1,598,194 1,295,738 2,893,932 1,446,966 1,446,966 Year 5 6,004,601 6,004,601 1,558,946 8,053 263,538 1,830,536 4,174,065 1,444,021 180,138 29,282 38,433 20,000 60,046 125,172 1,897,092 2,276,973 2,276,973 47,110 47,110 2,229,863 2,229,863 557,466 1,672,397 1,446,966 3,119,363 1,559,682 1,559,682 Year 6 Year 7 Year 8 6,304,831 6,620,072 6,951,076 6,304,831 6,620,072 6,951,076 1,700,898 8,858 289,892 1,999,648 4,305,183 1,516,222 189,145 32,210 42,276 20,000 63,048 113,963 1,976,864 2,328,319 2,328,319 2,328,319 2,328,319 582,080 1,746,239 1,559,682 3,305,921 1,652,960 1,652,960 1,856,349 9,744 318,881 2,184,974 4,435,099 1,592,034 198,602 35,431 46,503 20,000 66,201 103,809 2,062,580 2,372,518 2,372,518 2,372,518 2,372,518 593,130 1,779,389 1,652,960 3,432,349 1,716,175 1,716,175 2,026,613 10,718 350,769 2,388,100 4,562,976 1,671,635 208,532 38,974 51,154 20,000 69,511 94,609 2,154,416 2,408,560 2,408,560 2,408,560 2,408,560 602,140 1,806,420 1,716,175 3,522,595 1,761,297 1,761,297 Year 9 7,298,630 7,298,630 2,213,135 11,790 385,846 2,610,771 4,687,859 1,755,217 218,959 42,872 56,269 20,000 72,986 86,270 2,252,573 2,435,286 2,435,286 2,435,286 2,435,286 608,822 1,826,465 1,761,297 3,587,762 1,793,881 1,793,881 Year 10 7,663,561 7,663,561 2,417,502 12,969 424,431 2,854,901 4,808,660 1,842,978 229,907 47,159 61,896 20,000 76,636 78,709 2,357,284 2,451,376 2,451,376 2,451,376 2,451,376 612,844 1,838,532 1,793,881 3,632,413 1,816,207 1,816,207
23 PREF 51/March, 2008/2
14.2 Projected Balance Sheet

Year 0 Assets Current Assets Cash & Bank Pre-paid annual land lease Raw Material Inventory Stocks-RM Account Receivable Total Current Assets Fixed Assets Building/Infrastructure Net Value of Bulls Machinery and Equipment Total Fixed Assets Total Assets Liabilities & Equity Short Term Liabilities Accounts Payable Other Liabilities Long Term Debt Total Liability Equity Paid-up Capital Retained Earnings Total Equity Total Equity and Liability Year 1 Year 2 Year 3 Year 4 Year 5 Year 6 Year 7 Year 8 Year 9 Year 10
120,000 20,000 92,436 232,436 867,500 525,000 1,009,000 2,401,500 2,633,936
219,513 22,000 92,436 411,667 203,014 948,630 836,625 473,750 908,100 2,218,475 3,167,105
502,575 24,200 100,724 432,250 213,164 1,272,913 807,294 427,563 817,290 2,052,146 3,325,059
583,506 26,620 109,792 453,863 223,823 1,397,603 779,429 385,934 735,561 1,900,924 3,298,528
535,529 29,282 119,718 476,556 235,014 1,396,099 752,958 348,413 662,005 1,763,375 3,159,474
392,902 32,210 130,583 500,383 246,764 1,302,843 727,810 314,590 595,804 1,638,204 2,941,047
553,535 35,431 142,480 525,403 259,103 1,515,951 703,919 284,098 536,224 1,524,241 3,040,192
671,187 38,974 155,508 551,673 272,058 1,689,399 681,223 256,607 482,602 1,420,432 3,109,831
758,602 42,872 169,778 579,256 285,661 1,836,169 659,662 231,819 434,341 1,325,823 3,161,991
821,999 47,159 185,410 608,219 299,944 1,962,731 639,179 209,466 390,907 1,239,553 3,202,284
864,228 51,875 202,539 638,630 314,941 2,072,213 619,720 189,308 351,817 1,160,844 3,233,057
1,316,968 1,316,968 1,316,968 1,316,968 2,633,936
45,585 1,117,732 1,163,317 1,316,968 686,819 2,003,787 3,167,105
49,672 890,604 940,276 1,316,968 1,067,816 2,384,784 3,325,059
54,144 631,677 685,821 1,316,968 1,295,738 2,612,706 3,298,528
59,039 336,501 395,540 1,316,968 1,446,966 2,763,934 3,159,474
64,397 0 64,397 1,316,968 1,559,682 2,876,650 2,941,047
70,264 70,264 1,316,968 1,652,960 2,969,928 3,040,192
76,689 76,689 1,316,968 1,716,175 3,033,142 3,109,831
83,726 83,726 1,316,968 1,761,297 3,078,265 3,161,991
91,435 91,435 1,316,968 1,793,881 3,110,849 3,202,284
99,882 99,882 1,316,968 1,816,207 3,133,174 3,233,057
24 PREF 51/March, 2008/2
14.3 Projected Cash Flow Statement

Year 0 Operating Activities Net Profit Add: Depreciation Expenses Amortization Expenses Raw Material Inventory Finished Goods Inventory Account Receivable Accounts Payable Cash Provided by Operations Financing Activities Land lease expense Add: Land lease payment Change in Long Term Debt Issuance of Shares Cash Provided by/(Used for) Financing Activities Investing Activities Capital Expenditure Cash (Used for)/ Provided by Investing Activities Net Cash Cash Balance Brought Forward Cash Available for Appropriation Dividend Cash Carried Forward Year 1 Year 2 Year 3 Year 4 Year 5 Year 6 Year 7 Year 8 Year 9 Year 10 1,373,639 1,448,812 183,025 166,329 (92,436) (8,288) (411,667) (20,583) (203,014) (10,151) 45,585 4,087 (92,436) 987,568 1,580,206 20,000 (22,000) (199,236) (201,236) 786,333 120,000 906,333 686,819 219,513 22,000 (24,200) (227,129) (229,329) 1,350,878 219,513 1,570,391 1,067,816 502,575 1,523,661 1,598,194 1,672,397 151,222 137,549 125,172 (9,069) (9,925) (10,865) (21,613) (22,693) (23,828) (10,658) (11,191) (11,751) 4,472 4,895 5,358 1,638,015 1,696,828 1,756,483 24,200 (26,620) (258,927) (261,347) 1,376,669 502,575 1,879,244 1,295,738 583,506 26,620 (29,282) (295,176) (297,838) 1,398,990 583,506 1,982,495 1,446,966 535,529 29,282 (32,210) (336,501) (339,429) 1,417,054 535,529 1,952,584 1,559,682 392,902 1,746,239 1,779,389 1,806,420 113,963 103,809 94,609 (11,896) (13,028) (14,270) (25,019) (26,270) (27,584) (12,338) (12,955) (13,603) 5,867 6,425 7,037 1,816,815 1,837,369 1,852,610 32,210 (35,431) (3,221) 1,813,593 392,902 2,206,495 1,652,960 553,535 35,431 (38,974) (3,543) 1,833,826 553,535 2,387,361 1,716,175 671,187 38,974 (42,872) (3,897) 1,848,713 671,187 2,519,900 1,761,297 758,602 1,826,465 1,838,532 86,270 78,709 (15,633) (17,129) (28,963) (30,411) (14,283) (14,997) 7,709 8,447 1,861,565 1,863,151 42,872 (47,159) (4,287) 1,857,278 758,602 2,615,880 1,793,881 821,999 47,159 (51,875) (4,716) 1,858,435 821,999 2,680,434 1,816,207 864,228
(20,000) 1,316,968 1,316,968 2,613,936 (2,401,500) (2,401,500) 120,000 120,000 120,000
25 PREF 51/March, 2008/2
15 ASSUMPTIONS
Table 15-1 Economy Assumptions 10% 10% 10% 10% 10% 365 4 208 100 20,800 10 208,000 5% 197,600 5% 15 15 30 100,000 Rs 500 per bull 8,500 per month Rs. 15,000 per month 4% of Value of Animals 3% of Revenue 1% of Revenue Wages growth rate Raw material growth rate Electricity growth rate Equipment growth rate Promotional expense growth rate Table 15-2 Production Assumptions Operational days Semen ejaculate per bull per Week Semen ejaculate per bull per Year (52 week) Dozes per ejaculate Semen dozes per bull per Year Number of bulls Total semen dozes (Year 1) Dozes loss Semen dozes available for sale (Year 1) Sales price growth rate Table 15-3 Cash Flow Assumptions Accounts Receivable cycle (in days) Accounts Payable cycle (in days) Raw material inventory (in days) Minimum Cash balance Table 15-4 Expense Assumptions Vaccination & Medication (Annual) Chemical Expense (Annual) Electricity + Generator charges Insurance rate Promotion & Travelling Expense Miscellaneous Supplies
26 PREF 51/March, 2008/2
Table 15-5
Raw Material Assumptions 350 kg 10% of body weight of bull Rs 55 per 40 kg 3 kg per bull Rs 15 per kg 180,000 Rs 2 197,600 25 5% 10 years 50:50 5 1 14% Sole proprietor tax rate 25% 5% 10% 10% Reducing Balance Reducing Balance Reducing Balance
Body Weight (bull) Fodder requirement per bull Fodder Price Feed Requirement per bull Feed Price Liquid Nitrogen Refilling (Year 1) Plastic tubes for filling of Semen/per tube Table 15-6 Revenue Assumptions Semen dozes available for sale (Year 1) Sale price per dose (Rs) Sale price growth rate (Semen dose) Table 15-7 Financial Assumptions Project Life Debt/Equity ratio Debt Tenure (Years) Payments per Year Interest rate on long term loan Tax rate Discount rate for calculation of NPV Table 15-8 Depreciation Assumptions Building & Civil Works Equipment Bulls
27 PREF 51/March, 2008/2
ANNEXURE 15
PROTOCOLS FOR SEMEN COLLECTION, EVALUATION, PROCESSION, STORAGE, THAWING & INSEMINATION PROCEDURES SEMEN COLLECTION A: Preparation of Artificial Vagina (AV) Day before collection: Assemble AV properly. Check for any leakage. Cover the collection tube wit insulated bag. Store in incubator at 42O C overnight. Drain the extra water from the valve. Keep water up to valve level by tilting AV. Inflate AV with proper pressure. Lubricate AV with sterile (UV) jelly. Ideally, use separate AV for each ejaculate. Rewarm AV, if it is not used within 15 mins. Disassemble AV. Drain the water from AV through valve. Immerse hard casing and cone-in-running water. Brush the hard casing with soap and water. Rinse in warm water several times. Place them vertically in incubator (40O C) for drying.
On collection day:
After Semen Collection:
Preparation of Bull: Wash the body and prepuce thoroughly. Eyoke the sexual stimulation by: o Ten minutes restraint in the collection area. o Once false mount on teaser. Avoid the contact of penis with the body or tail of teaser. SEMEN EVALUATION Identify the collection tube by bull and by ejaculate. Place the tube in water bath (37OC) immediately. Cover it with rubber stopper.
5
Courtesy: Dr. Younas, Deputy Director, Qadarabad SPU. 28
Note the volume and record. Take one drop on clean slide, put cover slip on the drop and check sperm motility and record. Record the sperm concentration wit semen photometer or with haemocyto meter. Calculate the dilution ratio for 60x10^G spermatozoa / ml as follows: o {volume x (sperm motility 100) x sperm concentration} 60 x 10^6* o * The product of this calculation will be the dilution ratio between semen and extender. Add 100 ul of semen in 2 ml of formol saline (1% formaldehyde in saline solution) and mix it well. Put a drop of mixture on haemocytometer chamber under cover slip. Count five squares diagonally. Total sperm from five squares will be the concentration in million, e.g. Total sperm in 2 squares is 100, the concentration will be 100 x 10^6. > 0.5 ml > 50% > 400 x 10^6 < 15%
Determination of Sperm Concentration using Haemocytometer
STANDARDS FOR ACCEPTABLE EJACULATE Volume: Individual motility: Concentration: Sperm abnormalities: B: Semen Procession Semen Dilution: Place semen and extender in a water bath kept at 37OC. Hold semen at 37OC for 15 minutes (holding time). Determine the amount of extender to be added. First, add equal amount of extender to the semen, while shaking the bottle / tube gently, to avoid glycerol shock to spermatozoa. Add the remaining part of extender to semen while shaking the bottle / tube gently. Place the semen at room temperature (22O-25OC) or 10 min. Immerse the bottles / tubes in plastic beaker containing water (22O-25OC). Transfer the plastic beakers along with bottles / tubes containing extended semen in cold cabinet (4OC). Check the temperature after every 30 minutes. When temperature of water in beaker reaches below WC, prepare ice water (4OC) in a tub. Transfer the beakers in tub. Monitor the temperature of beaker, it will come down to 4OC within 30 min. When temperature reaches 4OC, note the time.
29 PREF 51/March, 2008/2
Semen Cooling:
C:
Let semen stays for 3 hrs after reaching at 4OC (equilibrium time = 3hrs) Semen Packing: Print the straw by bull and by date. Put the straw in small plastic bags. Sterilize them under UV light. After completion of equilibration time, fill the straws with extended semen of respective bull (one by one). While filling the straws of one bull, the semen of other bulls should be resting in the lower. Compartment of cold cabinet to avoid the re-warming of semen. Wile filling the straws, do not lift the lid of cold cabinet for longer time, it will re-warm the upper camber of cabinet. After filling, arrange the straws on racks and keep the racks at 4OC for 10 min. Semen Freezing: Turn on the freezer (see separate instructions). Stack the racks. Transfer the racks in Programmable Freezer and freeze the semen in two steps: o From 4OC to -15OC @ -3OC/min. o From -15OC to -80OC @ -10OC/min. Plunge the semen into liquid nitrogen. Distribute the semen in goblets, bull-wise. Store it into liquid nitrogen container. Post-thaw Semen Quality: Thaw semen straw at 37OC for 15 sec. Check the drop of semen for sperm motility. Store only those semen doses possessing more than 40% sperm motility and discard rest.
C:
C:
PREPARATION OF EXTENDER Composition of Extender: For 1 Liter Ingredients Tris (N-tris-hydroxy-methyl-amino methane) Citric acid, monohydrate Na-citrate dehydrate Fructose Egg yolk Glycerol Penicillin Streptomycin sulfate Double distilled water TCA 29.89 gm 15.79 gm 2 gm 200 ml 60 ml 1000 iu 100 ug 740 ml Citrate 29 gm 2 gm 200 ml 60 ml 1000 iu 100 ug 740 ml
30 PREF 51/March, 2008/2
Preparation of Extender: Weigh the buffer part of extender, i.e. Tris+citric acid in case of TCA extender, and Na-citrate dehydrate in case of Citrate extender and put it in sterilized 1000 ml glass cylinder. Weigh fructose and put in the same cylinder. Add distilled water slowly up to the level of 740 ml of cylinder. Cover the cylinder with tin foil or parafilm and stir it with magnetic stirrer for 15 minutes at room temperature. Pasteurize the cylinder containing buffer(s) and fructose at 65OC for 30 minutes. Cool the cylinder in running tap water till it reaches room temperature (22O25OC). Add egg yolk, glycerol, and antibiotics one by one while it is stirring. Label it with date and initials and store at 4OC for 2 overnights (to settle down the egg yolk particles). Transfer the supernatant to the flask and keep it in the refrigerator. The extender will be useful for 10 days.
SPERM AND ACROSOME MORPHOLOGY 1. Formal Saline Solution: a. Dissolve 3.4gm Sodium Citrate Dehydrate in 100ml distilled water. b. Remove 1ml from citrate solution. c. Add 1ml of Formaldehyde (37% commercial preparation) in citrate solution. 2. Sperm / Acrosome Morphology: a. Take 0.5ml Formal Saline solution in a glass vial. b. Add 1 2 drops of semen in the solution. c. Observe at lease 200 spermatozoa for head, midpiece and tail and acrosome abnormalities under phase contrast microscope.
31 PREF 51/March, 2008/2

Pre-Feasibility Study: Livestock Semen Production Unit

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Pre-Feasibility Study

LIVESTOCK SEMEN PRODUCTION UNIT

Small and Medium Enterprise Development Authority

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

Pakistan Economic Survey 2006-07 Pakistan Economic Survey 2006-07

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

Rationale and Justification of the Investment Opportunity

Agriculture Census Organization, Statistics Division, Govt. of Pakistan.

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

The cost of proposed project is approximately Rs 2.632 million.

CURRENT INDUSTRY STRUCTURE

There are four SPUs in the country at the following locations.

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

TECDOC-1480, IAEA International Atomic Energy Agency

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

HUMAN RESOURCE REQUIREMENT

Recommended Mode for Acquiring Land

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

Buffalo/ Cattle Bull 8. 4

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

11 KEY SUCCESS FACTORS

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

12 THREATS FOR THE BUSINESS

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

23 PREF 51/March, 2008/2

Livestock Semen Production Unit (Cattle & Buffalo)

14.2 Projected Balance Sheet

120,000 20,000 92,436 232,436 867,500 525,000 1,009,000 2,401,500 2,633,936