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MAHARASHTRA ANIMAL & FISHERY SCIENCES UNIVERSITY, NAGPUR.

COLLEGE OF VETERINARY AND ANIMAL SCIENCES UDGIR 413 517, DIST. LATUR.

REFRESHER TRAINING COURSE ON

ARTIFICIAL INSEMINATION

– 413 517, DIST. LATUR. REFRESHER TRAINING COURSE ON ARTIFICIAL INSEMINATION DEPARTMENT OF GYNAECOLOGY AND OBSTETRICS

DEPARTMENT OF GYNAECOLOGY AND OBSTETRICS

MAHARASHTRA ANIMAL & FISHERY SCIENCES, NAGPUR COLLEGE OF VETERINARY & ANIMAL SCIENCES, UDGIR- 413 517 DEPARTMENT OF GYNAECOLOGY & OBSTETRICS

REFRESHER TRAINING COURSE ON

G YNAECOLOGY & O BSTETRICS REFRESHER TRAINING COURSE ON Sponsored by Maharashtra Livestock Development Board, Akola

Sponsored by

Maharashtra Livestock Development Board, Akola

in collaboration with

Maharashtra State Animal Husbandry Department

with Maharashtra State Animal Husbandry Department   F IRST D AY TIME TOPIC FACULTY 10.00
with Maharashtra State Animal Husbandry Department   F IRST D AY TIME TOPIC FACULTY 10.00
with Maharashtra State Animal Husbandry Department   F IRST D AY TIME TOPIC FACULTY 10.00
 

FIRST DAY

TIME

TOPIC

FACULTY

10.00

hrs

Registration

11.00

hrs

Inauguration session

12.00

hrs

Introduction to AI course

Dr. N.M. Markandeya

12.30

hrs

Pre training evaluation

13.00

hrs

Lunch break

14.00

hrs

Structural dynamics of reproductive tract

Dr. R.R. Mugale Dr. S.S. Kulkarni Dr. N.Z. Gaikwad Dr. M.A. Khan

14.45

hrs

Follicular dynamics and oestrus cycle

15.30

hrs

Oestrus related behaviour & Heat detection

16.15

hrs

Morbid specimen palpation

17.00

hrs

Participants presentation

 

SECOND DAY

TIME

TOPIC

FACULTY

09.00

hrs

Library Access

10.00

hrs

Physiology of oestrus & hormonal control

Dr. N.M. Markandeya Dr. A.V. Bhosale Dr. S.D. Moregaonkar

11.00

hrs

Infectious infertility & it‟s control

12.00

hrs

Uterine pathology and conception rate

13.00

hrs

Lunch Break

14.00

hrs

Artificial insemination technique

Dr. A.D. Patil Dr. S.D. Moregaonkar Dr. N.M. Markandeya Dr. B.S. Borgaonkar

14.45

hrs

Strategies to improve conception rate

15.30

hrs

Sexual health control in males and females

16.15

hrs

Management information system

17.00

hrs

Group discussions on Gynaeco-clinical problems Dr. N.M. Markandeya Dr. A.D. Patil

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THIRD DAY

TIME

TOPIC

FACULTY

09.00

hrs

Climatology & Reproduction

10.00

hrs

Cattle and Buffalo breeding activities

Genetic improvement of cattle through AI

Fertility improvement measures

Dr. S.S. Kulkarni Dr. P.B. Deshpande Dr. M.P. Sawane Dr.N.M. Markandeya

11.00

hrs

12.00

hrs

13.00

hrs

Lunch Break

14.00

hrs

Scientific feeding & ration computing

Dr. A.B. Kanduri

14.45

hrs

Role of nutrients on fertility

Dr. A.U. Bhikane

16.00

hrs

Common infertility problems in bovines

Dr. A.D. Patil

16.45

hrs

Post thaw semen evaluation

17.30

hrs

Discussions on surgical problems

Dr. S.B. Daware Dr. V.D. Aher Dr. G.U. Yadav

 

FOURTH DAY

 

TIME

TOPIC

FACULTY

09.00

hrs

Library access

10.00

hrs

Induction & Synchronization of oestrus

Post partum disorders & infertility

Pharmacodynamics of drug molecules

Dr. A.D. Patil Dr. N.M. Markandeya Dr. G.D. Ranvir

11.00

hrs

12.00

hrs

13.00

hrs

Lunch Break

14.00

hrs

Accidents during insemination

14.45

hrs

Health monitoring for high conception rate

Cold chain management of frozen semen

Dr. G.U.Yadav Dr. B.N. Ambore Dr. P.B. Deshpande

15.30

hrs

16.15

hrs

Handling & management of LN 2 containers Dr. N.M. Markandeya

17.00

hrs

Discussions on clinical cases

Dr. A.U. Bhikane Dr. B.N. Ambore

 

FIFTH DAY

TIME

TOPIC

FACULTY

09.00

hrs

Internet & Information systems

Prof. C.R. Joshi Dr. A. D. Patil Dr. N.M. Markandeya Dr. A.A. Devangare Dr. J.M. Chahande

10.00

hrs

Follow up of AI

10.45

hrs

Modern trends in improving breeding efficiency

11.30

hrs

Clean milk production, recording and testing

12.15

hrs

Economics of dairying

13.00

hrs

Lunch Break

14.00

hrs

Post training evaluation

15.00

hrs

Valedictory session

16.00

hrs

Participants remarks

17.00

hrs

Relieving reports

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OTHER ACTIVITIES

Guest faculty lectures

Demo of AI films

Clinical case discussions One theme debates

On farm demonstrations

Clinical practices

Company presentations

PROGRAMME ORGANIZATION COMMITTEE

Dr. N.M. Markandeya, Programme Co-ordinator

Dr. A.U. Bhikane, Technical arrangements

Dr. N.Z. Gaikwad, Lodging arrangements

Dr. S.B. Daware, Registration arrangements

Dr. A.D. Patil, Organizing Secretary

SUPPORTING FACULTY

Dr. S.D. Moregaonkar, Guest arrangements.

Dr. G.P. Bharkad ,Boarding arrangements.

Dr. G.U. Yadav, Audio visual arrangements.

Dr. B.N. Ambore, Refreshment arrangements.

Dr. S.T. Kalwaghe, Transport arrangements.

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Sr.

   

No.

List of Topics

Name of Faculty

   

Dr. N. M. Markandeya

1

High conception rate through artificial insemination

Dr. A. D. Patil

   

Dr. N. M. Markandeya

2

Insemination Technique in Bovines

Dr. A. D. Patil

   

Dr. N.M. Markandeya

3

Nutritional infertility in bovines

Dr. A.U. Bhikane

4

Reproductive Nutrition in Bovines

Dr. A. B. Kanduri

   

Dr. P. B. Deshpande

5

Retrospective Review of Breeding Strategies for Livestock

Dr. M.N. Sawaisonkamble

   

Dr. B. N. Ambore

6

Health Monitoring For High Conception Rate

Dr. A. U. Bhikane

7

Induction of Estrus In Anestrous Animals

Dr. Mukund Amle

8

Handling And Management Of LN 2 Containers

Dr. S.C. Vora

9

Infertility in Bovines

Dr. S. C. Vora

10

Morbid uterus palpation and identification of different structures

Dr. Mukund Amle

11

Evaluation of Semen

Dr. Mukund Amle

12

Record keeping in Artificial Insemination

Dr. S. C. Vora

13

Follicular Dynamics And Oestrus Cycle

Dr. S.S. Kulkarni

14

Climatology And Reproduction

Dr. S.S. Kulkarni

15

Inflammatory Conditions Of The Uterus

Dr. S. D. Moregaonkar

4

Sr.

   

No.

List of Topics

Name of Faculty

16

Pharmacodynamic of Drug Molecule

Dr. G.D.Ranvir

17

Clean Milk Production and Testing

Dr. A. A. Devangare

18

Structural Dynamics of Female Reproductive Tract

Dr. R. R. Mugale

19

Oestrous Related Behaviour And Heat Detection

Dr. N. Z. Gaikwad

20

Infectious Infertility And Its Control

Dr. A. V. Bhonsle

21

Therapeutic management of anoestrus in farm animals

Dr. N M Markandeya

 

Need of Re-emphasizing Extension Activities of Artificial Insemination

Dr. N M Markandeya

22

Dr. S. D. Moregaonkar

23

Abortions in Dairy Cows: New Insights and Economic Impact

Dr. S.B. Barbuddhe

24

Health Management for Optimum Production and Reproductive Performance in Dairy animals

Dr. E. B. Chakurkar

25

Diagnosis of Tuberculosis, John‟s Disease and Sexuality transmissible diseases (STDs) In Bovines

Dr. S. S.Rautmare

26

Accidents During Artificial Insemination : Be Careful

Dr. G. U. Yadav

27

Non-inflammatory Conditions of Uterus

Dr. S. D. Moregaonkar

28

Strict Hygiene and Sterilization for Artificial Insemination

Dr. A. V. Bhonsle

29

Post Thaw semen evaluation

Dr. S.B. Daware

   

Dr. A. U. Bhikane

30

Role of Minerals and Vitamins in Reproduction

Dr. B. N. Ambore

31

Genetic Improvement of Indian Cattle

Dr. M. P. Sawane

5

High conception rate through artificial insemination

Dr. N M Markandeya and Dr. A D Patil Veterinary College, Udgir 413 517 Artificial insemination technology has been proved beneficial in the world and realizing benefits in terms of increased milk productivity and breeding efficiency cent percent animal population is being bred through AI in number of countries. Artificial insemination has made male factor constant in animal reproduction. However, unless carried out as per the physiological norms, artificial insemination may not give expected conception rates. Joint front of herd owner, semen bank and inseminator with frantic and earnest efforts can achieve the desired conception rate. Services per conception (S/C), non return rate, pregnancy rate, calving rate and herd reproductive status (HRS) are the important parameters to monitor female fertility and are considered as measures of reproductive efficiency in dairy animals. Conception rate is the proportion of cows that failed to return to service by thirty days after the end of the month in which they were inseminated. Ideal conception rate is expected as 1.6 inseminations per conception. Proper heat detection, Good quality semen, proper handling of semen, sound reproductive health of the cow, strict hygienic procedure and timely AI at appropriate site in cervical region of females in oestrus helps to achieve high fertility with artificial insemination and very high conception rate. Inseminator‟s role to collect the straw from container, proper thawing and loading of the gun with deposition of semen in female reproductive tract is vital. Every animal in oestrus should be attended most perfectly during the attempt of artificial insemination. The prerequisite : Raising of farmer‟s awareness regarding the technology is most important prerequisite in artificial insemination for high conception rate at field level Our livestock population is reared under traditional practices. Vast majority cattle and buffaloes still remain uncovered under artificial insemination services in nook and corner of the country. Actual number of pregnancies obtained through artificial insemination is very few. Use of single biotechnique of artificial insemination with all other traditional and routine husbandry practices limits the role of the highly effective technology. Hence, optimizing and advising technical advances in animal management systems is an essential pre-requisite for extending artificial insemination services to field level. For improvement of female fertility,

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artificial insemination needs to be coined with sexual health programme for improved milk productivity. There is common consensus amongst animal owners that natural service gives better conception rates than inseminations. But there is no method of breeding which gives cent per cent conceptions. Multifaceted advantages of AI should be made understood by the animal owners only then AI can be popularized. Farmers are interested in impregnating their animals without much consideration and thought on method of breeding as either artificial insemination or natural service. Improvement of farmer‟s awareness for success of the technology is also important. Educating farmers and to make them understand the benefits of technology is the most difficult task for veterinarians at field level and veterinarians should accept the challenge for rural development. Factors affecting conception rate Heat detection : Onus of heat detection is always with animal owner. AI is useful under the conditions, which consists of regular and prompt heat detection at farmer‟s level. In absence of heat detection, animals are not reported for AI. Untimely reporting or delayed reporting also limits expected conception rate. Time of insemination : Trained veterinarian is the best judge to decide the correct time of insemination. Fertilization failures are expected due to non-maturation or aging of gametes with asynchrony of their union. Timely inseminations with synchronous ovulation leads to higher conceptions rates and hence timely inseminations are to be designed by the inseminators. Number of Inseminations: Two or more number of AI attempts to suit in cases of ovulation delay may improve conception rate. However it should be an exceptional attempt in problem cases only as double inseminations are not practicable, possible and also economical. Generally, efforts should be made to achieve conception through single insemination. Stress effect of insemination: If disturbed at time of AI, poor chances of conceptions are expected in the animal. Avoid struggling of animal at the time of AI as adrenaline ACTH gets secreted and it blocks forward movement of sperms. Excitement of animals should be avoided before during and even after insemination. Noise, vehicular traffic, beating, dragging, tightly fixing with crate creates stress to the animal. Any known or unknown factor leading to stress lowers conceptions. Animals under stress or nervous stage during oestrus fail to conceive. Insemination after calving. : Optimum time for resumption of ovarian cyclicity coincides with complete uterine repair at cellular level. Insemination in animals before proper establishment of

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cyclicity fails to succeed. First post partum cycle may be defective as majority of animals are in negative energy balance and hence too early insemination leads to lower conception rate. Efficiency of inseminator. : It is universally accepted premise that only trained inseminators should carry out artificial insemination, who fully understand pros and cons of the technique. The inseminator should have thorough knowledge of physiopathology of reproduction. The training, efficacy, experience, ability, attitude and approach of the inseminator plays pivotal role in the success of artificial insemination programme. Precision and perfection of inseminators are the important prerequisites for maximum and consistent results of artificial inseminations. Regular refreshment of technical advances helps to improve efficiency. There is no specific time and site of artificial insemination within reasonable limits. The skill, which surely turns into conceptions, is only the best approach of any individual inseminator. Inseminator should standardize his own technical and practical skill for improving pregnancy rate instead of following blind steps of other colleagues. Unattended cases of inseminations presented on Sundays and holidays also reduce conception rates. Rejection of unfit cases: It is most practicable and clinical approach to refrain inseminations during all abnormal and pathological oestrual stages, instead of attempting it for owner‟s satisfaction. Sound health is indispensable for reproduction and hence animals with poor boy conditions should be rejected for inseminations. Owner should recognize the fact the every animal in oestrus may not be fit for insemination. All animals selected for inseminations must be in physiological, normal oestrus. Refusal of unfit cases for insemination improves the conception rate. Ovulatory delay. : Under non-infectious, cyclic and non-breeder category, this is the major problem in animals. Ovulatory delay extends stage of oestrus and also leads to aging of capacitated sperms. On delayed ovulation, egg degenerates due to non-fertilization and hence the conception rate is reduced. Diagnosis of expected time of ovulation and inseminating the animal at appropriate time is key to achieve ideal conception rate. Use of hormones: Exogenous hormonal injections improve conception rate through prompt ovulation, better development of CL, increase in level of progesterone and protection of conceptus during the period of ovum. Commonly gonadotrophines releasing hormones are widely used in clinical practice for improvement of conception rate. However, any approach involving progesterone therapy for a longer period than natural luteal phase used for induction of oestrus will reduce the conception rate.

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Environmental effect: Conception rates are suppressed during hot season. Milking cows are more severely affected than heifers. Variations and constant fluctuations in uterine temperatures decrease survival of conceptus in uterus. Nutritional effect: High plane of nutrition is essential for reproduction. Inadequate nutritive factors alter microenvironment at ampulary and endometrial region leading to misconceptions. Unusual and unsuitable feedstuff or estrogenic plants lower conceptions as their major effect is on process of sperm transport in female reproductive tract. Thus optimum provision of all nutrients is prerequisite for successful conceptions. Tips to improve conception rate:

Check and effect regular assessment of frozen semen straws. 2. Maintain cold chain, follow refilling of LN 2 timely. 3. Handle biological containers carefully with avoiding shocks and mechanical jolts. 4. Follow proper thawing procedure. The standard temperature of 37 0 C for 30 seconds is recommended. Both temperature and the time are important to achieve high recovery rate. 5. Protect artificial insemination gun carefully after loading till its use. 6.Select females for AI carefully by thorough gynaeco-clinical investigations. 6. Improve self-efficiency for heat detection in animals, timely AI, proper selection of site for semen deposition. 7. Be gentle while handling ovaries of animals in oestrus. 8. Follow strict sterilization procedure before and during insemination. 9. Do not inseminate during abnormal oestrus and pathological discharges. 10.Avoid inseminations to first pubertal oestrus or first post partum oestrus. 11.There is no fixed time and site for insemination. 12.Questionable semen quality leads to low conception rate. 13. Joint front can improve the conceptions. 14.There is no effect of volume of semen inseminated on conception rate. 15. Assess health score, soiling of perineum, mucus adhered to tail and thighs if any before gynaeco-clinical examination of the animal. It is not important to analyse what procedure is being followed at field level but it is important to guide on standard practices and procedure of AI for high conception rate It is always recommended to the inseminators to follow the procedure, which proves high conception rate and it is necessary to avoid the procedures which lowers the conception rate. In conclusion, it can be stated that artificial insemination is the national high way towards prosperity of rural poor. Oestrus leading to successful conception and subsequent calving resulting into milk production is expected at regular and ideal intervals. If the veterinarians accept the task of cent percent artificial insemination programme as a service to nation, no child will sleep without a cup of milk nor any Indian kitchen will remain without milk, which will in turn reduce starvation,

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child mortality and many more social problems. Let us reorient ourselves to impregnate oestrual cases through artificial insemination.

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Insemination Technique in Bovines

Dr. N M Markandeya and Dr. A D Patil

Veterinary College, Udgir 413 517

Artificial insemination is a technique in which high quality semen is deposited in the female reproductive tract as per physiological laws. Proper time of insemination, optimum dose i.e. Number of live motile spermatozoa and appropriate site of insemination are the important and highly essential prerequisites of successful artificial insemination. Every animal presented for insemination is entitled to best attention and efforts of the inseminator Time of insemination 1. Animals first noticed in heat in morning hours (A.M.) should be inseminated on the same day in evening (P.M.) and animals first noticed in heat in evening (P.M) should be inseminated on next day morning (A.M.). 2.Too early or too late insemination leads to repeat breeding. 3 Time of ovulation and time of insemination must be synchronized for successful fertilization. 4. Fixing correct time of insemination depends on inseminator‟s diagnosis, ovarian activity and expected duration of the oestrus phase. Non-descript cows and buffaloes remain in oestrus for short period of 12 to 18 hours. Descript cows; well-fed cows are generally in oestrus for 24 hours. Cress-bred cows and exotic cows show oestrus extended upto 36 hours and even more. Problem breeders have delayed oestrus phase of 48 to 72 hours. Buffaloes are generally in heat during night hours. There is no precisely defined „best‟ time for AI. Under these circumstances, correct insemination time solely depend as per the judgment of the inseminator and is always final. Always confirm fern pattern before selecting animal for AI, which confirms the heat /oestrus stage. The ferning examination helps to achieve upto 70% and 60% conception rate in cattle and buffaloes respectively. Number of sperms 1. Minimum 12 million to 15 million live sperms are required for successful conception. 2. At least 35 to 40 per cent motile sperm percentage is expected in thawed straw.3.Although only single sperm is required for fertilization, minimum concentration of sperms is necessary at the site of fertilization. 4. Sample straw check is necessary at regular monthly intervals and records the observations of the stock. 5. Irrespective of volume of semen and type of straw, optimum number of sperms per insemination is important. Oestrus stage is always susceptible for infection and therefore WBCs are present in the uterus. One leukocyte engulfs three sperms i.e. spermiophagy. Only live sperms come in contact with uterus as compared to dead sperms. Therefore, optimum sperm concentration is essential in

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the straw. Additionally, concentration of hyaluronidase enzyme can be achieved only after having presence of optimum number of sperms at the site of fertilization. Less than optimum number of sperms in live and motile state defeats very purpose of insemination. Insemination site 1. Intra-vaginal: more semen quantity is required and sperm loss is more, hence not used for artificial insemination. 2.Intracervical: Mid-cervical site is most commonly recommended site for insemination. 3. Intrauterine: Can be practiced, however it limits the role of cervix to discard dead, non motile an abnormal sperms. Deposition of semen at the level of body of uterus is more appropriate than inter corneal insemination. The AI is a bio-technique and should be followed with scientific standard procedure and as per physiological laws. Animals presented for inseminations are to be attended only after a short rest in the clinic as generally animals are brought from distance. Following steps should be used to carry out inseminations. Recording of breeding history: 1. Record detail breeding history of the animal. 2. Check the breeding sheet and data recorded by the farmer breeder, if any. 3. Check details of treatment adapted to the animal during recent past. 4. Avoid inseminations during first pubertal and first post partum oestrus. 5. Avoid too early breeding in heifers. 6. Also avoid animals exhibiting post partum oestrus before 60 days of calving. 7. Check inter-oestrus intervals, record of repeat inseminations and the dates of earlier inseminations. 8. Enquire the history and symptoms of the oestrus with time of detection. Gynaeco clinical examination: 1.Check external signs of oestrus and record changes in external genitalia. 2. Per rectal examination is mandatory before each insemination for confirmation of oestrus. 3. Check possibility of gestational oestrus, early pregnancy, mid cycle oestrus. 4. Check follicular developments on the ovaries. 5. Confirm the oestrus stage and decide the appropriate time of insemination in each case. Select animals for insemination on body score condition. Do not select late heats for inseminations. Never incline to owner‟s requests for inseminating unfit animals. Veterinarian‟s role is most important for rejection of unfit animals for insemination. Carrying out AI is rather easy but to diagnose whether cow is fit for insemination is difficult. Technical skill and trained human resource is required to select animals for insemination. Preparation of animal 1. Restrain the animal in cattle chute / crate / Travis properly. 2. Thoroughly clean and wipe external genitalia. 3. Ensure cleanliness and use ample antiseptic to

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clean external genitalia. Complete drying should follow 4. Cleaning. 5. Fix the tail on lateral side. 6. Assistance of one person is useful to monitor the animal. Type of frozen semen straw: Only frozen semen straws, which are now being used for inseminations in both cows and buffaloes. French straw ( Cassou method) consists of 12 cm long straw with 0.5cc capacity and it consists of one end sealed with cotton plug. German straw (Landshut method) consists of 6.5-cm. long semen straws with 0.30cc capacity and carries both ends sealed by metal balls. Thawing of frozen straw the procedure of thawing frozen semen dose of either German (Landshut) mini straws or French (Cassou) medium straws remains the same. 1. Dip the forceps in LN 2 container for pre-cooling. 2. Lift the canister upto the level of neck of the container. 3. Collect and grasp only one frozen semen straw at a time. 4. Never lift the canister to high above the level of LN 2 in any case. 5. Put the neck plug of the LN 2 container after properly placing the canister. 6. Air dry the straw for a single moment. 7. Dip the straw completely in lukewarm water for 30 seconds.8. Temperature of water used for thawing of frozen straw should be 35 to 37 0 C. 9. Thaw one straw at a time. 10. Thawing container should be of wide mouth type and also should accommodate the straw horizontally. Loading of AI gun : 1. Select proper AI gun as per the semen being used. 21. Lift the straw with forceps from thawed water and wipe of the water from surface with tissue paper. 3. Hold the straw vertically and adjust the air bubble towards the end from which the straw has to be cut. 4. Cut the straw in the middle of the air bubble at right angle. 5. Load the AI gun with the cut end of the straw at the forward point of gun. 6 Use sterilized AI sheath only and never reuse any sheath. 7. Wrap the loaded gun with clean dry paper, towel to protect the same from dust, dirt and UV sun rays with rains. 8. Remember pre-warming of gun and sheath is also necessary during cooler days. Insemination proper : 1. Frozen thawed semen is deposited in female reproductive tract by recto- vaginal method. 2. Pass left hand in the rectum and complete back racking. 3.Dilate the vulval lips with assistance of helper. 4.Insert AI gun at an angle of 45 0 s with orientation towards cervix. 5. Never touch the anterior portion of AI gun to perineum or any object. 6. The portion of AI gun should not come in contact with vestibular passage. 7.Make the AI gun horizontal and orient the tip of AI gun towards os cervix. 8.Guide the tip of AI gun by left hand through rectum and insert the gun into cervix. 9. Slowly pass the gun in os 10. Deposit the semen at mid cervical portion gently. 11 Remove AI gun very slowly and allow it to glide on the clitoris.

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Clitoral massage is helpful to bring about ovulation. Cooling of perineum with water can be practiced. Do not disturb the cow during insemination. Deliberateness and gentleness is highly essential. The cow should not recognize slight pain during insemination. Post AI care : 1. Always examine AI gun on removal for detecting blood tinged mucus, pus, and back flow of semen. 2. Record the date of AI, bull used, straw details on animal sheet. 3. Discard the used plastic sheath and straw properly. Advise to Animal owner 1. Keep the animal at doorstep and withheld grazing just for the period of oestrus cessation so as to avoid service by stray bull. 2. Provide ample water and feed as per daily needs of the animal. 3. Be gentle with the nervous animal during the period of oestrus. 4. Observe and record the period / time of cessation of oestrus signs. 5. Allow the animal to relax and even to sit in the shed. 6.Do not follow the quacks and mis-beliefs. 7.Observe met-oestrus bleeding, if any. 8. Observe the animal on 21 st , 42 nd day of insemination for repeat oestrus, if any. 10. Advise to present the animal for pregnancy confirmation after 60 days of AI. Points to be considered:1. Do not expose the semen straw / AI gun to direct sun or rain. 2. Do not touch the frozen straw with bare hands / fingers without forceps. 3. Shake the straw in air to remove excess LN 2 from surface. 4. Do not put the straw back into canister once exposed to the air. 5. Use the straw after thawing within 5 minutes only. 6. Handle the insemination tube with light touch. 7. There is nothing to be „prodded‟ out of the way or poked through with the AI gun. 8. Gentleness and deliberateness are essential. 9. While insemination stand like a fencer. Vaccinations and injections are totally different than inseminations. No un-trained person should be allowed to inseminate the animals. Guidelines and directives for inseminations can be followed but uniform rule could not be applied to the insemination procedure in all the animals. Be bold for scientific norms. Extend the technology of artificial insemination in qualitative terms with best possible conception rate. Mission to cover all breedable Indian animals under AI technology is only possible through joint front of skilled professionals.

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Nutritional Infertility In Bovines

Dr. N. M. Markandeya and Dr. A.U. Bhikane Veterinary College, Udgir 413517 Every known food factor is indispensable for normal reproduction. Malnutrition leads to lowered vitality, body resistance and fertility. Many practicing vets are convinced that there exist profound effect of nutrients on reproduction and but very few are able to determine the exact nutritional cause of reduced fertility. Very little is known about how nutritional deficiencies or their excess cause in fertility. They may act via hypothalamus and pituitary or directly influence on gonads thus influencing steriodogenesis and endocrine function. However, herd reproductive health and profitability may suffer, if there is underlying nutritional problem leading lowered fertility. Different physiological processes have different priorities in receiving nutrients. Depending on age and physiological status of the animal, nutrients are preferentially partitioned for lactation, growth and fattening. Reproductive functions have a relatively low and last priority for nutrient partitioning. Thus, in some instances and especially during scarcity, sub-clinical deficiencies may manifest themselves as impaired reproduction. Requirements for normal reproductive function in male are considerably less than in female. In addition, the added demands of lactation are not present, so borderline nutritional efficiencies are less likely to contribute to decreased male fertility. In females, there are several time periods in which nutritional deficiencies are more likely to affect reproduction. They correspond to times when metabolic demands or other physiological process are greatest, yet reproductive function is also at a critical time period. These phases include the period of rapid growth during puberty, parturition and peak lactation with resumption of cyclicity. Peak lactation is the time when it is very difficult to meet the nutrient requirements of the dairy cow and to impose on her the necessity of having a fertile oestrus and conceiving. First calf heifers that are still growing and they have an additional demand. The major 99.99 per cent animal matter is made up form four fundamental elements as H, Co, N 2 , O 2 and 7 micro minerals Viz. Calcium, Phosphorus, Sulfur, Potassium, Sodium, Chlorine, and Magnesium. These elements exist as water, fat, protein and carbohydrates. 13. Energy effect: Energy is required for body cell functioning, maintenance of body temperature, secretary processes and formation of new tissues.

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Of the energy supplied through ration, consumption and priorities of the same is generally fixed. As 30 per cent goes unutilized waste in feces, 5 per cent is passed out through ruminal gases, 5 per cent is cleared through urine and 20 per cent is utilized for digestion, only 40 per cent net energy remains for body processes. The energy is basically used for other physiological processes like metabolism and growth followed by lactation and lastly for reproduction. Energy levels have significant effect on ovarian activity as low energy than optimum level leads first to ovarian inactivity. Deficient energy intake is the most significant nutritional factor in anoestrus. The most severe effect of inadequate nutrition is cessation of oestrus cyclicity and other less severe conditions are silent oestrus, ovulatory defects, conception failure and early embryonic mortality. Sub-maintenance feeding results in prolonged oestrus cycles, cessation of oestrus, anovulation or ovulation without oestrus. Low progesterone concentrations are related with the low energy levels. Trophoblastic proteins and or endometrial secretary proteins might form biochemical dialogue between mother and the conceptus that is essential for embryo survival. Progesterone level is responsible for the production of these proteins. However, high plane of nutrition during early pregnancy is associated with low conception. Energy, which can be stored in body as carbohydrates is very limited and is sufficient for just one day. Excess carbohydrates are converted and stored as fat. Fat obesity due to excess feeding leads to fat deposition around ovaries and bursa, which interferes normal follicular developments, ovulation and ovum transport. Fat deposition in endometrium also hinders implantation. Pubertal stage : Energy deficiency signs thin animal, inactive ovaries, delay in puberty. The onset of puberty is closely related with energy in male and females and body weight is most important factor. Effects of underfeeding are more pronounced when applied in the early post-natal than prepubertal. Housing and management of breeding age heifers are frequently less than optimum and also there exists deficiency of energy in diet. During growth stage, adipose issue has lower priority than lean tissue and nutrient restrictions may delay the acquisition of a minimum level of fatness desired for the onset of puberty. Plane of nutrition delays puberty by inhibiting the pulsatile release of GnRH from hypothalamus and consequently of LH from pituitary. Since increase in pulsatile LH secretion

16

stimulates the development of ovarian follicles to preovulatory surge, its interruption due to under nutrition might be the primary mechanism in delaying the onset of puberty. Heifers on lower energy diet had lower serum progesterone concentration and elevated LH concentration, suggesting that LH release was not impaired but there may be decreased ovarian response to LH with restricted dietary intake. Substances that mediate the effects of nutrition on LHRH relieve (through CNS) have not been identified but neurons controlling LHRH release are sensitive to availability of metabolic fuels. The circulating concentrations of insulin, growth hormone, certain amino acids, non- esterified fatty acids and others act as signals of nutritional status. Monensin, an ionophore commonly used as a feed additive in beef cattle rations to improve average daily gain and feed efficiency decreases the age of onset of puberty in beef heifers. Monensin supplementation increases the GnRH and estrogen induced LH response in prepubertal heifers, which may be related to its effects on puberty. Once cyclicity has commenced at puberty, it should continue uninterrupted throughout the animal‟s life apart form pregnancy stages and feeding level does not appear to affect cyclicity unless severe restrictions occur. Short term under nutrition has beneficial effect on conception rate, which increases progesterone level required for pregnancy maintenance. NRC energy requirements should not be exceeded on prepubertal heifers. Animals that are fed excessively before and around time of puberty appear to have inadequate mammary secretary tissue development and lower subsequent milk production. Heifers fed a high plane of nutrition have increased mammary fat deposition and decreased secretary tissue at prepubertal age. Heifer is under stress in post pubertal period because she is growing to physical maturity and also conceiving with pregnancy maintenance. There is no evidence that growth restrictions in early life will influence reproductive performance once the feeding of normal diet has been implemented. Liberal or high energy feeding early in life hastens onset of sperm production in male animals, but moderate restrictions after two years of age are desirable for diet to avoid leg weakness. Postpartum period Parturition results in abrupt shifts in metabolic demands from nutrients, which are generally spared from body reserve. Proteins and lipids stores are metabolized to support sudden milk production. Rapid increase in energy requirements for milk production results in

17

negative energy balance. The stage starts a week before calving and reaches to severity in two weeks of lactation. Dairy cows in early lactation draw upon their body energy reserves to produce large volumes of milk. Because maximum feed intake lags behind peak milk yield by several weeks, in early lactation it is nearly impossible to achieve adequate energy intake to sustain production. Production is maintained by using body reserve, primarily fat. Thus, there is considerable weight loss during early lactation. Commonly, veterinarians and owners consider negative association between high production and fertility. Many studies have demonstrated no direct relationship between high production and reduced fertility. Fertility, progesterone concentration and energy balance are directly related with each other. Provision of supplemental fat in the diet has beneficial effect on limiting energy imbalance during post partum period and also on improvement of conception rate. As consistent increase in plasma cholesterol levels will stimulate steroid synthesis and body health status. It is important to maximize dry matter intake of high energy feed early in lactation, both for maximum milk production and best reproductive performance. Post partum period should have high energy feed to avoid weight loss. Dairy cows gaining weights are more likely to conceive. Similarly, maximum energy intake should not extend beyond peak lactation, because this may lead to over conditioning of the cow with severe consequences to health and fertility. Energy intake should be increased up to peak milk yield and then decline with normal decline in production. Excessive energy intake will result in fattening during late lactation and dry period. Excess grain consumption has been reported to have adverse effects on fertility in several studies. Over conditioned cows are predisposed to develop fat cow syndrome. Fatty infiltration of liver leads to reduced fertility. High quality forage availability is essential to maximize dry matter and energy intake. Energy status has been frequently thought to be in direct relation with post-partum fertility. During increased susceptibility of early post partum period, variety of peri-parturient diseases will adversely effect on fertility. B. Protein effect : Proteins are required for maintenance, growth, lactation and reproduction. The effects of protein nutrition on reproductive process might be via the action of specific amino acids that are needed for the synthesis of GnRH neuro- transmitter. Rations that provide adequate protein

18

for maintenance and growth are also adequate for reproduction. The level of energy is more important for reproduction than protein. Common deficiency is related with protein, phosphorus and vitamin A level in the diet. Protein deficiency results in a delay in onset of puberty, increased days open, decreased dry matter intake. Decreased dietary intakes are due to poor digestibility of ration. Readily available proteins can furnish ammonia for growth and multiplication of rumen microbes. Proteins digested by small intestines, which is furnished by a combination of microbial protein synthesized in rumen and protein that is not readily available to ruminal degradation. The amount and composition of the protein reaching the small intestines determines the productive capacity of cow in terms of protein supplementation. Urea and non-protein nitrogen (NPN) sources furnish ammonia that the rumen organisms can synthesis into protein. Normally, 11 to 12 per cent crude protein levels are required to sustain adequate rumen ammonia for normal rumen fermentation, digestion and dry matter intake. These levels are adequate for normal reproduction in bulls, heifer and dry cows. Higher levels are required for growth and lactation. Soybean can be used in animal feed as readily available protein source. Very high or above the normal requirements of protein concentrations have negative effects on fertility. 17 to 18 per cent crude proteins will increase production but is undesirable for reproductive efficiency. Excess feeding of dietary proteins increase rumen ammonia, increase blood urea, increase open days, increased services per conception and toxic effects on embryo. Urea is a highly soluble source of ammonia and has been used commonly as protein source in ruminants. NRC recommendation 1% of dry matter and slow ruminal adoption is also important. Urea is a highly soluble source of ammonia and has been commonly recommended as protein source to the ruminants. As per NRC recommendations, use of one per cent dry matter with slow ruminal adoption of urea will be helpful. When feeding higher levels of protein or NPN, it is best to provide some by pass protein so that rumen ammonia levels do not become excessive. C. Mineral effect: Soil is not only the principal source of trace elements entering the food chain but is also a sink for elements from environmental sources. Minerals are required for reproduction because of their cellular roles in metabolism, maintenance and growth. Minerals have specific roles and requirements in reproductive tissues and the role and requirement may change as per the physiological state of cyclicity or pregnancy. Mineral imbalances are thought to be associated with infertility.

19

Inorganic or mineral elements also called as major / macro minerals consists of calcium, phosphorus, potassium, sodium, chlorine, magnesium and sulfur. Micro minerals or trace element consists of iron, iodine, zinc, copper, manganese, cobalt molybdenum, selenium boron, lithium, bromine and vanadium. Role of fluorine, chromium, tin, silicon, nickel and arsenic is not yet proved. Most of the cases of reduced fertility are usually due to multiple mineral deficiencies. Mineral requirements are too less in ruminants and their availabilities are also very small in the feed. Indiscriminate and regular use of chemical fertilizers in agriculture during last 50 years has totally imbalanced soil- plant- blood constitution of mineral status. Diagnosis of mineral deficiencies is difficult as their signs are rarely pronounced and also because estimations are costly. Calcium deficiency does not cause reproductive failure but Ca: P ratio matters much. Hypocalcaemia limits absorption of other minerals. Hypocalcaemia leads to milk fever, dystocia, retained placenta, metritis and delayed uterine involution. Phosphorus is most frequently associated with reproductive abnormalities. Hyperposphatemia D. Vitamin effect: Vitamins are supplied either through diet or they are synthesized in the body by ruminal microbes and even by tissues. Depletion of feed source due to long storage, non-exposure to sunlight limiting tissue synthesis and stressful conditions lead to vitamin deficiencies. Adverse effect of vit A occurs during latter half of gestation as abortion, birth of dead calf, dystocia, placental retention. Three doses of Vit. A therapy is recommended during mid gestation and after parturition if green fodder is not available or animal has to pass through stressful summer. Vitamin B is synthesized in bovine rumen and hence deficiencies are less likely to occur. Adrenal cortex and corpus luteum contain high concentration of ascorbic acid. Vit E and vit C are related with hormone productions but vit C is not reported to disturb reproduction. Vit D is supplied in sun cured forages, grasses and is also synthesized from cholesterol in skin upon exposure to sunlight but vit D has no direct role on reproduction. Antisterility factor vit E acts with selenium and functions as antioxidant. Feeding sprouted oil seed is highly recommended for fertility improvements. Borderline nutritive deficiencies are manifested as impaired fertility before other clinical symptoms are apparent. To identify or prevent nutritional problems, it is important that an effective working relationship exists between the dairyman, veterinarian, nutritionist and feed salesman. It is better to prevent nutritional deficiencies rather than to treat it.

20

Reproductive Nutrition in Bovines

Dr. A. B. Kanduri Veterinary College, Udgir

Introduction: - In reproducing animals, as in growing animals there is an interaction between nutrition and

production. The influence of nutrition on reproduction begins early in the animal‟s life, as the plane

of nutrition in young animals can affect the age at which they reach puberty. In mature animals,

poor nutrition can reduce the production of ova and spermatozoa, so that the female either fails to

conceive or produce fewer offspring than normal. Generally much emphasis should be given on the

role of energy intake (general plane of nutrition) in reproduction, because deficiencies or excesses

of specific nutrients often affect reproduction through their influence on energy intake. In cattle the

main problem of fertility is that of obtaining reconception in cows two months after calving, at a

time when the nutritional demands of lactation are high and must often be met partly from the body

fat reserves. In dairy cows in New York state, over a period of 17 years when milk yield increased

by 33 %, the proportion of cows conceiving to their first insemination fell from 66 to 55%.

Requirement of energy and protein during growth & puberty: -

Puberty in cattle is markedly influenced by the level of nutrition at which animals have

been reared. In general terms, the faster an animal grows, the earlier it reaches sexual maturity.

This is illustrated in following table.

Age & size at puberty of Holstein cattle reared on different planes of nutrition.

Plane of Nutrition

(Percent

accepted standard

for TDN)

At Puberty

Sex

of

Age (Weeks)

Weight (Kg)

Female a

High (129)

Medium (93)

Low (61)

37

49

72

270

271

241

Male b

High (150)

Medium (100)

Low (60)

37

43

51

292

262

236

a : Sorenson A M et al. 1959 Bull. Cornell Univ. Agric. Exp. Stn, No.936. b : Bratton R W et al. 1959 Bull. Cornell Univ. Agric. Exp. Stn, No.940.

21

This table shows the effect of three planes of nutrition on the initiation of reproductive ability in dairy cattle. Although in both sexes there were considerable differences in age at puberty between the three treatments, difference in live weight were much smaller. In female animals the primary determinant of fertility (i.e. weather or not animal conceives) is the number of ova shed from the ovaries. In case of cattle the rate is normally one. It has been suggested that high plane of nutrition increases ovulation rate by stimulating the pituitary gland to produce more of the hormone involved in ovulation, leutinizing hormone. Another endocrinological explanation is that a high plane of nutrition promotes a greater production of insulin, which encourages the uptake of glucose and the synthesis of steroid hormone by the ovary. In mammals, the spermatozoa, ova and the secretions associated with them represent only very small quantities of matter. The average ejaculate of the bull, for example, contains 0.5 gm of dry matter. It therefore seems reasonable to suppose that nutrient requirement for production of spermatozoa and ova are likely to be inappreciable compared with the requirements for maintenance and for process such as growth and lactation. An adult male animal kept only for semen production would require no more than a maintenance ration appropriate to their species and size. But in practice such animals are given food well in excess of that required for maintenance in females of the same weight. There is no reliable evidence that high planes of nutrition are beneficial for male fertility, though it is recognized that underfeeding has deleterious effects. After mating the plane of nutrition should be reduced to about the maintenance level. Higher levels of feeding after mating appear to lead to losses of ova by stimulating the metabolism (i.e. destruction) of progesterone, the hormone required for the establishment and maintenance of pregnancy. Protein deficiency can be expected to influence reproduction through its effect on feed intake. However, it has recently been suggested that a supplement of digestible protein that is undegradable in the rumen (DUP) can increase the ovulation rate in cattle. It has also been suggested that an excess of rumen degradable protein (RDP) in the diet of dairy cows can cause ammonia toxicity, which in turn reduces fertility, as expressed by an increase in the number of inseminations required to secure conception. Also a prolonged protein deficiency especially in young animals leads to reproductive failure. The production of ova and spermatozoa follow the attainment of sexual maturity (puberty), and undernutrition during growing period delays puberty in both males and females of domestic animals. This undernutrition during the growing period also affects the production of eggs and of

22

sperm after puberty. Generally amounts that are adequate for maintenance will meet requirements

for ova and sperm production.

Semen characteristics of Bulls Receiving 100, 120, and 140% 0f Recommended Maintenance Requirement.

 

Semen Characteristics

 

Feeding level

Volume (ml)

Motility (%)

Sperm x 10 3 (Per mm 3 )

100% of maintenance

5.70

72.7

1219

120% of maintenance

5.64

73.4

1226

140% of maintenance

6.50

73.2

1220

[C. Branton, R.W. Bratton and G.W. Salisbury, Journal of Dairy Science. 30:1003-1013,1947.]

Low energy intake alone has been reported to markedly delay puberty, although in some

studies, energy was not the only dietary variable. High-energy intakes have also been shown to

affect reproduction. Too much energy may cause so much fat as to interfere with follicle

development on the ovaries in the female and with sperm production in the male, along with

decreased desire to mate in both. While at least part of the problem with low energy may be the

effect on hormone production, low protein intakes have been clearly shown to affect hormone

production. Reduced gonadotropic and sex hormones resulting from low protein intakes in turn

affect development of the sex organs, desire to mate, and production of sperm and ova. Reduced

hypophysial gonadotrophines are the major cause of reproductive malfunction in protein

deprivation. The leutinizing hormone seems especially sensitive.

Energy & Protein balance during Pre and Post-partum: -

Good milk production and numbers of calves per unit time are only obtained by achieving

early conception in heifers and a short inter-calving interval in adult cows. Energy is an important

nutrient for dairy cows both before and after calving and there is no substitute for energy in the diet

of ruminants. A balance of energy and protein is required, even before calving and in the dry

period. Energy deficiency before calving (below maintenance) should be avoided as well because

this leads already at this stage to metabolic stress and low conception rates in the following

lactation.

The negative effect of an insufficient energy provision before calving will be enhanced by

an energy deficiency after the following parturition. The importance of energy after parturition is

well known. Already in the first two to three weeks of lactation, energy from any source is

23

important for the onset of ovarian and, related to this, for uterine involution. Energy deficiency

leads to acyclia, silent heat, delayed ovulations and follicular cysts. Significant correlations exist

between fertility and weight loss or body condition, as indicators of negative energy balance in the

first weeks after calving.

Results in pregnancy, number of treatments and inseminations per pregnancy in cows with different nutritional status and health.

 

Nutritional status/health

Energy deficiency liver damage

 

Parameter

 

Normal

Energy deficiency excess of protein

% of pregnancies

 

94.4

81.0

54.5

Treatments per cow

 

1.22

1.46

1.66

Inseminations/pregnancy

1.39

2.19

2.59

Interval

treatment

conception

48.50

66.90

68.30

(days)

(Escherich and Lotthamer, 1987)

Cows gaining weight during early lactation have a higher conception rate and need fewer

services per conception compared to those losing weight.

Effects of a cow’s condition on conception rate

 

Cow’s

Total

Number

Total

Number

 

Conception

Condition

Services

Pregnant

Services/Conception

Rate

Gaining Wt.

1368

911

1.50

67%

Losing Wt.

544

234

2.32

44%

Cows that lose an excessive amount of body condition or fat stores during early lactation

have longer intervals to first ovulation and first estrus (heat period), lower first service conception

rates and more days open.

Improvements in a cow‟s energy balance may be an important signal to the ovaries to start

cycling. Preliminary results from research trials indicate that energy balance may also influence

developing ova (eggs). When energy status becomes more positive, early post partum, diameter of

the largest follicle on day 10 post partum, increased double ovulation, increase the day of detection

of the first Corpus Luteum (CL) was earlier. These changes, thought to be aroused by increases in

LH-FSH, insulin, IGF, and other yet to be determined compounds as activated by improving

energy status. In order to avoid large negative energy, energy balance, and maximize DMI, well-

24

known conventional management practices are recommended. Supplemental fat may increase energy status thereby improving follicular recruitment and growth. Fat supplement have been shown to increase cholesterol, precursor of progesterone which may increase fertility. Protein deficiencies in lactating cows may increase the incidence of silent heats (cow releases the egg but she is not seen in heat) and lower conception rates while at the same time decreasing feed intake and milk production. Excesses of protein (crude protein in the total diet greater than 17-20%) have been implicated in lowering conception rates with increases seen in the number of services per conception and days open. When an excess of degradable protein and/or a deficiency of energy is fed, ammonia not incorporated into microbial protein is absorbed into the blood stream. In turn, this excess ammonia and urea in the blood stream may decrease fertility at the same time energy is diverted away from milk production and/or reproduction. Some studies have indicated that blood urea nitrogen (BUN) above 20 mg/100 ml may decrease the chances of pregnancy. Recommends dietary concentration of 19% during the first 3 weeks post partum and 16- 18% thereafter, depending upon the amount of milk being produced. The reason for higher recommendation (19%) just after calving is low DMI. During the first 10 weeks post partum, cows also undergo tremendous changes as they shift from pregnant to non-pregnant states and prepare to conceive again. The pituitary and hypothalamus undergo changes in control of metabolic and reproductive hormones, uterus undergoes morphological and histological changes, and the ovary undergoes recrudescence, i.e. out-break activity after a period of inactively of follicle recruitment and growth leading to ovulation and formulation of CL. Breeding often initiated at 8-10 weeks post partum under the influences of this dynamic metabolic scenario. If more protein is consumed that can be effectively utilized, urea (primarily) and ammonia increase in body fluids despite increases in hepatic concentrations or urea cycle enzymes. Urea concentration in blood (BUN) was increased on average 2.54 gm/100 ml for each pound of dietary CP intake. Mechanisms by which these negative effects occur are not clear. Concentration of ammonia/urea or another unknown nitrogenous compound can be sufficiently high in the body tissue to hamper fertilization, embryo development, and implantation of the conceptus, thereby retarding genesis of a new calf. Some studies show that P4 concentrations in cows fed 15-20% CP were lower than cows fed 13% CP. Feeding high amounts of protein increases concentrations of nitrogenous compounds in blood. Urea has proved toxic to ova and sperm.

25

Retrospective Review of Breeding Strategies for Livestock

Dr. P. B. Deshpande,(Deonikar) Asstt.Commissioner of A.H. Dist.A.I.Centre, Latur.

Dr.M. N. Sawaisonkamble, Regional Joint Commissioner of Animal Husbandry, Latur.

Britishers adopted horse breeding for military purposes. However mixed farming has been

a practice of farmers since preindependance. Cattle were reared mainly for draft power, milk and

manure.

Upto 4 th Five Year Plan, bull rearing was the concept as bullock power constituted major

agricultural requirement.

Milk production acquired top priority to satisfy urban demand of cheaper milk to urban

population. Developed countries also played their roll in pouring funds for mechanization of dairy

sector to liquidate their over productions.

Animal Husbandry Department of Maharashtra tried to preserve indigenous breeds by

posting them in rural areas for natural services till 1960-65. Schemes like registration of Deoni,

Khillar and Gavlav cattle were implemented in respective regional pockets of breed. However, due

to increasing demands of milk, the crossbreeding was introduced from 1967 onwards in certain

areas.

In accordance with the policy laid down in 1981, the crossbreed of N.D.cows with H.F.was

taken up in irrigated tracts and use of Jersey bull semen was introduced in non-irrigated tracts and

drought prone areas. Up gradation of local buffalo was done by Murrah/Mehasana buffalo a bull in

irrigated areas and Surati was used for non-irrigated/ draught prone areas. The policy delivered six

time rise of milk from 1062 thousand MT in 1970-71 to 6376 thousand MT in 2003-04. The share

of buffalo milk is to the extent of 46 % in 2003-04.

Apart from crossbreeding, the areas were also identified for preservation of Indigenous

germ plasm as per statement enclosed. The official inseminators are expected to inseminate or

breed indigenous cattles with respective breeding bulls only. The farmers have also a liberty to

adopt individual breeding plan for his personal needs.

Now the Govt.of India has announced a national policy on agriculture in July 2000. As a

sequel to this, National Livestock Policy has been identified and saline features of policy are

1. Stabilization of livestock population in respect of land load and its balance with growing

demands of livestock products.

26

2.

Low productive potentials in bovine population, poor coverage of A.I. and alarming

conception rates associated with feeds and fodder shortages.

3. Negligible grazing resources.

4. Livestock environment, Improvement of soil fertility through mannuring.

5. Livestock and human hygiene, hygienic production of livestock products.

Improvement of buffalo may not be concentrated on one or two defined breeds but a

biodiversity be maintained.

6. Creation of special zones for rearing of bullock/ bulls for sustainable breed conservation

7. Encouragement of breeders associations

Salient features in Animals Husbandry & dairy sector in last few year will explain how the

farmers in Maharashtra are responding to programme which are planned with their help, for their

development.

The roll of veterinarian is to stand as bridge between planning visionaries and end

beneficiaries that is farmers.

Annex

Breed

District

Tahasils

Deoni

Latur

Ahamedpur, Udgir, Chakur.

Lal Khandhari

Nanded

Khandhar , Mukhed, Deglur, Ahamedpur, Loha, Biloli, Nanded.

Gaolao

Wardha

Arvi, Karanga, Kharangana, Ashti.

Dangi

Ahamednagar

Akole.

Nashik

Igatpuri.

 

Solapur

South Solapur, North Solapur, Mohol,Malshiras,Karmala, Sangola, Manglveda, Pandhapur ,Akkalkot,Madha,Barshi.

Khillar

Satara

Man khandala, Khatav, Sangli.

Sangili

Miraj, Kavate Mahankal, Jath,Athpadi,Khanapur ,Vita.

Pandhapuri

Solapurs

South Solapur,North Solapur, Akkalkot,Mohol,Barshi, Pandhapur , Manglveda, Malshiras.

Nagpuri/

Wardha

Ashti, Karanga, Arvi.

Amravati

Anjnggaon-surji, Chanaur Bazar.

Achalpuri

Akola

Akot.

State Govt also reviewed its policy on the backdrop of National policy and adopted revised

breeding policy from Feb.2004 as per its G.R.No.LVS.102003 /Page No.467.ADF-4, dt.7 th

Feb.2004.

27

The policy changed is done considering increasing human population and its corresponding demand of livestock products. The need of employment generation to educated unemployed and necessity of preservation of pure and famous indigenous breeds. The vital roll of M.L.D.B. also compelled Govt. this decision. The important features of policy are as under: - A- 1- The use of exotic jersey frozen semen is to be made on non-descript cattle.

13.

The use of exotic H.F. frozen semen is to be made in areas with high irrigation facilities for agriculture and with those who have skilled management practice and sufficient green fodder.

3-

Exotic level is to be strictly restricted to 50% only.

4-

The limit of exotic level can be extended to 62-5 in areas, which are awakened in Animal Husbandry and management fields with permission of Department and Govt. The cows produced by adopting this policy will be called as MAHANANDINI and bulls as MAHANAND.

13.

With a view to preserve indigenous breeds, the treasures of Maharashtra like Dangi, Khillar, Deoni, Lal Kandhari and Gavlav be produced in their defined home tracts by

adopting scientific and advanced technology. C- 1- Breeding of Nagpuri and Pandharpuri be done in original home tracts by supplying frozen semen of proven bulls.

13.

Non-descript buffalo be bred with Murrah and Surati.

13.

Murrah and Mahsana purebred frozen semen doses be utilized in urban and semi-urban area.

13.

The other rural area should be covered with Surati and Murrah breeds.

It will be binding to adopt this policy to Government and Government aided institutes and individuals. If any organization or individual wants to deviate, can do so on the responsibility and risk, however, the permission of Government to do so is essential. They are also required to see that no damage is made to original indigenous breeds.

28

Breeding policies of Government of India/Maharashtra (Results as viewed by the authors)

Milk production/Draft/ Etc Military purpose served Milk 831TMT Milk production 1062 TMT

Sr.

No.

Period

Policy

Results

1

Pre-

Independence

Natural Breeding of horses for military purposes

Quality up gradation achieved Start of Indian policy. Home breeds paid attention less attention to milk production.

2

Post

Independence up to 1970-

71

Natural and artificial technique in breeding. Propagation of traditional home breeds. Cross breeding on specific military farms. Natural breeding by posting indigenous bulls with farmers and village Panchayat. Cross breeding of ND cows with HF in green zones and Jersey in Dry zones. Emphasis on enhancement of milk production attention towards up gradation of buffalo with Murrah and Surti. Use of Jersey in Dry areas and HF in irrigated. Strict adherence of 50% exotic level. Escalation up to 62.5% allowed in developed areas. Preservation of Indian breeds in specified Tahsils.

3

1981-2003

Inception of scientific approach. High milk production arena looked.

Milk 2003 TMT (1982 1983) Milk 6376 TMT

4

2004

onwards

More scientific approach. Creation of milk zones. Biodiversity maintenance angles.

Results awaited

29

Health Monitoring For High Conception Rate

Dr. B. N. Ambore & Dr. A. U. Bhikane Veterinary College, Udgir

To enhance maximum possible production and reproduction, health status of the animal

should be well. There are several factors that cause a departure from the state of health interrupting

the performance of vital functions. Among these vital functions of the body one of the most

important is conception, the beginning of life, get impaired if the general health of the animal is not

optimum. The following factors should be considered to ensure the health status and indirectly the

reproductive processes.

1. MANAGEMENT:

A scientific animal husbandry practices should be followed to overcome the reproductive

problems. These include

a. Hygienic conditions Hygienic conditions should be maintained at every stage of management.

b. Environmental conditions :

Heat during summer months causes decreased thyroid activity, which may lead to reduced

reproductive performance.

Cold excessive exposure to cold may cause failure of oestrus cycle in young growing

animals

Humidity.

c. Production stress / physiological stress High milk producing dairy animals may not conceive

early / easy following parturition due to negative energy balance.

d. Transportation stress Long distance transportation stress leads to oestrus failure due to

absence of regressing corpus luteum and hormonal disturbances.

2. NUTRITION :

Nutrition is one of the important factor which influences reproduction usually by failure of

oestrus, cessation of oestrus and sometimes failure of conception / early embryonic death.

a. Overfeeding Overfeeding leads to fattening of animal and in the overfat animal ovaries

are small and also fat deposition in the ovaries interferes normal ovulation and transport of

ovum. The extra fat in the ovaries also obstructs the development of follicles. While in the

bulls also overfat interferes with sperm production and libido.

30

b. Underfeeding Underfeeding results into delayed sexual maturity in heifers and also cessation of oestrus cycle in all aged animals. Underfeeding may leads to deficiency of energy, protein, phosphorus and vit-A. When there is low energy, follicular atresia develops (failure to mature the follicles).

Hypoglycaemia at oestrus may affect the conception due to the lack of energy to spermatozoa / fertilized ova, therefore all the breedable animals should be fed by adopting the standard feeding practices so that they should be in a positive energy balance after calving to realize the best conception rate.

c. Vitamin deficiency Any vitamin deficiency as such is not associated with infertility, but deficiency of vitamins may result in dystocia, abortion / still birth and also the advance effect on the ovarian cycle.

d. Mineral deficiency Along with all other vital functions of the body minerals also plays

an important role in improving fertility and life time efficiency of dairy animals, particularly the deficiency of phosphorus, copper, iron, cobalt, iodine and selenium have more influence on fertility. Vitamin E and selenium: Helps in transportation of the spermatozoa to the site of fertilization by improving uterine contractibility and Vit. E is also involved in synthesis of reproductive hormones i.e. progesterone and estrogen.

3. COMPTON METABOLIC PROFILE TEST:

The laboratory evaluation of certain blood components on regular basis will reflect the nutritional status of the animal.

We can assess the qualitative and quantitative adequacy of the diet of cows expected to produce

a certain quantity of milk and also to return the animals to estrus within the desirable length of time following parturition.

is a reliable test for early diagnosis of nutritional deficiency/metabolic imbalance.

is also useful for the selection of superior animals.

Thus by doing C.M.P.T. of herd, we can very well assess the nutritional demand of dairy animals and indirectly the productive and reproductive performance of them.

It

It

31

Metabolic Diseases and Conception:

Among the metabolic diseases P.P.H. (post parturient haemoglobinuria) and ketosis have an indirect effect on conception and fertility of animal. In PPH due to phosphorus deficiency and in ketosis due to negative energy balance and hormonal imbalance of insulin and cortisol. 4. HEALTH CONTROL MEASURES:

To maintain the general health of animal the following practices should be routinely adopted.

A. Deworming- Due to favorable climatic conditions i.e. hot and humid environment of majority

states of our country, gastrointestinal parasitism is more common which causes the nutritional deficiencies as well as anoestrus. Animals with heavy worm load can show the clinical signs but

those with low grade of infestation may go unnoticed. In most of the cases without knowing the primary cause we administer hormonal therapy for induction of oestrus. In many of cases worm load may be the primary cause of anoestrus. The vitamin and mineral deficiencies and heavy helmintic infestation are the major cause of anoestrus in dairy animals. The gastrointestinal parasites are responsible for poor digestion and improper assimilation of digested food resulting in nutritional deficiency. It has been also recorded that the parasitic infestation also reduces the serum calcium and inorganic phosphorus. The parasitic worms also causes the decreased serum iron by blood sucking activity. Thus the intestinal parasitism may lead to nutritional deficiencies along with macro and micro mineral deficiency and which may lead to infertility, for that purpose the regular anthelmintic treatment is necessary. Principles to be followed to control parasitic diseases:

Herd & its management

● Shed/ house management

Nutritional status

● Control of vector population

● Pasture management ● Protection of young animals

Deworming Schedule

● Age of first deworming – 3 days.

● Regular- thrice a year in animals above 6 months of age.

● then at monthly interval upto 6 months.

B. Spraying - The external parasites also causes the severe harmful effects on animal health.

The ecto-parasites suck the blood, which may lead to anaemia.

Severe itching of skin leads to restlessness and thus reduced appetite and water intake.

32

Some ticks are responsible for transmission of the protozoan diseases such as babesiosis, theileriosis etc.

Loss of body weight in all animals. All these effects of ectoparasites may impair the reproductive performance of the animals. Therefore regular insecticide spraying on animal body as well as in the animal shed is essential.

C. Vaccination So many infectious diseases are responsible to affect the reproductive performance of dairy animals. All these diseases cause the abortions in different stages of gestation. The cows those abort before 5 months of gestation hardly shows the retention of placenta but those aborts after 5 months, retention of placenta is most common and which later complicates the estrus and conception. Following are the common diseases that causes abortion

A. Bacterial-

Most common - Least common -

B. Viral-

Brucellosis, Leptospirosis, Tuberculosis, Vibriosis Haemorrhagic septicaemia, Anthrax, Salmonellosis, Listeriosis.

Most common

IBR, FMD

Least common

BVD, RP, MCF

C. Protozoan-

Babesiosis, Trypanosomiasis, Anaplasmosis, Toxoplasmosis Trichomoniasis

D. Mycotic/fungal Aspergilosis

Principles to be followed to control infectious diseases:

To avoid the infection of these diseases following control measures should be adopted

Maintenance of hygiene and sanitation.

Infected animal should be isolated/ sold / slaughtered.

Eliminate the source in infection

Aborted foetus / placenta should be deeply buried

Disinfections of infected area/houses.

Prevent genital contact between infected and noninfected animals. (Adopt A.I.).

Avoid stress due to transportation, inclement weather, undernutrition

Regular screening of herd for various tests like MRT, Tuberculin test etc.

Regular vaccination.

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Induction of Estrus In Anestrous Animals

Dr. Mukund Amle Veterinary College, Shirwal.

ESTRUS (Estrum): It is fairly well defined period characterized by sexual desire and the

acceptance of the male by the female domestic animals.

COWS: -

The intensity of estrus is weak, medium or intense. Majority of cows come in heat at night

& is detected early in the morning. Cow in heat exhibit partial in appetence due to excitement. She

prefer standing in byre, restlessness, nervousness moves anxiously in the field, not interested in

grazing, as the cow approaches estrus, she tends to sniff & lick the perineum of other cows. She

will access the receptivity of other cows by resting her chin on the rump or lion of other cows. She

arches her back and lumbar region during heat. Tail is slightly raised. The hairs of the tail head or

often ruffled &the skin some times excoriated by mounting by other cows. Bellowing is feature of

heat, marked drop in milk yield, frequent attempts of micturition. Rumination is slightly decreased

during heat.

PHYSIOLOGICAL CHANGES:

1. Vulva & Vagina: - In estrus vulva is congested, edematous, turgid, wrinkles on vulva disappears,

and stands predominantly growth and thickening of vaginal epithelium becomes hyperemic and

congested. Vaginal PH is 7.32

2. Cervix: - Cervical and vaginal glands secrete increased amount of mucous (mucin). Cervix is

relaxed, alteration in physiochemical properties of cervical mucous. The genital discharge of

transparent mucous whose elasticity causes to hang in complete clear strands from the vulva to

the ground is called as bulling string. Mucous adhere the tail and flank of the cow in heat.

Viscosity of the mucous is decreased, ferning is maximum on mucous dried on glass slide in

the beginning, the oestrual discharge is thin, shiny, clear, odorless and hanging out. During

estrous, mucous is stringy, adhesive, transparent and copious and occasionally containing

exfoliated cells.

3. Uterus: - Erect, turgid, edematous. Uterine tone is maximal on rectal palpation. Endometrium

becomes hyperemic and congested.

4. Ovaries: - Usually 1 to 5 small follicles start growing but undergo atresia. During estrus one

follicle grows & ovulate. Ovarian follicle is smooth convex fluctuating glister like measuring

about 0.5inches.

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5.

Oviduct: - Tonic epithelium matures cilia active oviduct contracts fimbriated ends of are assuming close affinity to the graffian follicle in the ovary.

oviduct

HORMONAL CHANGES: - During proestrus, the preovulatory follicles secret increasing amount of Oestradial. At this stage cow may ride on other cows and begins to secretes cervical mucous. Basal level of milk or plasma progesterone is reproductive confirmation of estrus. At the time of onset of estrus, pick level of Oestradial trigger a source of LH that causes ovulation to occur about 10-12 hr. after the end of estrus in cow. 0.5-0.80c raises body temperature during ovulation and drops at ovulation by about 0.3 0 c. METHODS OF ESTRUS DETECTION 13. Per rectal examination: - To be undertaken by technician

2. Standing heat: - The best indication of estrus is when the female stands when mounted by male

or by other females. Cows are observed in loose house early in the morning & evening every day.

They are observed for standing heat when mounted by other cows. The non-pregnant cows are watched for 20 to 30-min.each times for standing heat. Videotapes are also placed in the loose room with video monitors and recorders.

3. Teaser bull / Androginised cows: - Teasers are surgically prepared bulls to prevent the release of

sperm or copulation or steers are treated with testosterone hormone or cow with cystic ovaries are also used as teasers.

4. Chin ball-mating device: - A halters is fitted with a reservoir of dye to the teaser that released is

by the ball type mechanism that marks a line on the back of cow in heat. Chin ball marker leave on

ink mark upon the ridden cows indicating that standing & acceptance to mount have occurred.

5. Grease: - Grease is smeared on the brisket of the teaser cow.

6. Pressure detection on the cow: -The detectors are applied on point of maximum pressure during

monitoring particularly sacral spine. Her pressure sensitive devices change the colors when animal

stands for mounting, also called as heat mount detectors. 13. Estrus related odors (pheromones): - Dogs are train to detected the odors in vaginal mucous, milk or urine of cow .a specific and distinct odor is present in the urogenital tract of cow during estrus the odor is apparently disappear or greatly attenuated during diestrous. Results indicate that dog can be trained to detect and respond to the odor associated with estrous in cattle.

35

8. KaMar heat mount detectors: - they are more expensive and cow must be identified when they are displaced during riding by other cows. 13. Closed Circuit Television with time lapsed video is quite effective.

10. Milk Progesterone assay: - Low progesterone level (0.02ng/ml) indicates that cow

For a short time after service to the cow, cow stands with raised tail and arched back and when such posture is adopted, it indicates that mating has occurred. Within tow days of natural service there is occasional yellowish white discharge of mucus containing neutrophils and the leukocytes from the uterus (heat period 12-14 hrs). BUFFALO: - Acceptance male is the more reliable sign of estrus in buffaloes. Discharge of clear mucus from vulva, vulval edema, restlessness, frequent urination, vocalization, sniffing and licking of genitalia of other animals, and drop in milk production are not reliable sign of estrus in buffaloes. Estrus commences towards late evening with a peak sexual activity between 6 am and 6 pm. EWES: - Estrus period last for 24 to36 hrs in ewes. Estrus in ewes is relatively inconspicuous and is not evident in absence of ram, hence the estrus is difficult to detect. In estrus ewes are restless seek out the ram and together for „harem‟ (courtship). The vulva may be edematous and mucus discharge from the vagina is scanty. The ewe may isolate her self from the rest of flock and waggles her tail and moves it laterally. Ewes show a preference for the ram of their own breed or a particular ram. Standing of the ewes is most easily noticed sign of estrus. Ewes do not exhibit homosexual behavior but this may occasionally seen in a does. During estrus, irregular cornification of the surface cells of vagina is noticed. Without presence of male, however estrus is difficult to detect in both ewes and does. Ram paws with forefeet, rubs his head along ewe side, nips her wool. Ram mounts and makes series probing pelvic thrust and than dismount. The introduction of ram to ewes during the transition from anoestrus season, stimulate then to come in heat and ovulate. Vasectomised rams are commonly used to detect the estrus ewes with making crayons, colored grease, and paint applied to the brisket of ram. The number of services received by an estrus ewe average at least about 4. Ram may serve 8 to 38 ewes in a day. DOES: - Estrus duration last for 24 to 48 hrs. Sign of estrus are more conspicuous in does than ewes. A doe in estrus in restless bleats frequently waggles her tail constantly and rapidly. She may have

in estrus.

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reduced appetite decrease in milk production. Swelling and reddening of vulva in conjugation with rapid wagging of tail and vocalization. Mucus discharge is scanty. Doe may occasionally exhibit homosexual behavior but not the ewes. Without the presence of male, estrus is difficult to detect. Buck show interest and will follow does 3 to 5 days before standing estrus occurs. This suggests a proestrus activity in does. Goat‟s odor from the giants located in the back of head between the horns MARES: - Estrous lasts for 7 days. Mares in estrus becomes restless and irritable, licks mare or stallion. During estrus vulva becomes large and swollen, labial folds are loose and open on examination labia is orange in a colour, wet and glossy and covered with a film of transparent mucous. Vaginal mucosa is highly vascular and thin watery, mucoid discharge may accumulate in vagina.

During diestrus only finger can be inserted in cervix. Mares assume status characteristics of urination called as micturition posture. Urine is expelled in small amount at several successions voids urine with repeated exposure of clitoris. Tall, had is often raised. Clitoris is erected producing a typical sound called as wrinkling of vulva. Mares in estrus leans her hind quarters, spreads her hind legs, lowers her pelvis, raise her tail to side. She takes a considerable interest in courtship. At the same time she is aggressive and prone to bite or kick the stallion when approached. Courtship between mare and stallion during estrus is aggressive and violent she shows vicious temperament. Mares are receptive to stallion from 3-4 days depending upon duration of estrus. Stallion usually exhibits “Flehman” reaction when mare is in heat. Mare housed in a special teaser paddocks. Indication of acceptance of stallion by mare are elevation of tail, spreading of legs, standing, frequent urination and counteraction of vulva. SOW : - Estrus lasts for 40-72 hrs. Onset of estrus is characterized by gradual changes in behavioral pattern. Sows in heat seek the boar when he is with in the site and assume rigid stationary stands and lardosis. Ears of sow become erect in response .Sow exhibit Muzzling action. She is restless and mouth on other animals. Peculiar repeated sow beginning 3 day before heat emits grunt. Vulva becomes swollen and congested and these features persist. Reddening of surrounding region. Cervix is rigid and edematous, occasional mucous discharge, which is thin, vaginal epithelium lining are thicker and keratinised. Estrus could be readily determined by pressing loin of sow with the palms of both hands. Estrus sow will stand motionless with cocked ears. Immobilization

37

responded by sow can be elicited if attendant sits on sow by producing the voice of boar sound site and of vasectomised boar are helpful to detect estrus. Boar odour is released by salivary glands. Skin glands also produce an odour. Male secretion of preputial pouch give an odour involved in sexual carpel glands is well differentiated in both the sexes producing specialized. Sexual receptivity lasts on an average of 40-60 hrs. Ova released 38-42 hrs after the onset of estrus. The sow seeks boar in proestrus. She nozzles his testicles and flank and may mount him. BITCH :- Estrus lasts for 9 days and ovulation occurs between 2-4 days after commencement of heat. Indication of estrus precedes the commencement of bleeding for several days. A day also before the end of proestrus the attitude of bitch changes. She shows signs of courtship towards male become restless. Seeks male for mating. During estrus she becomes stationary She allows male stands, Display vulva by arching her and deviate and held her tail one side. Vulva swelling and tummification are greatest on onset of stage of estrus. Bloody discharge from vagina also watery, reddish or yellow as heat proceeds. During proestrus she will accepts the male. The period of intense desire is during the first 2-3 days of estrus, during which copulatry tie occurs lasting for 15-25 min. during proestrus and estrus there is proliferation of vaginal epithelium, which undergoes stratification or cornification. In the height of estrus there is shedding of keratinisation surface of all vaginal epithelium. Since pheromone release is maximal at this stage, roaming bitches are often followed by a pack of clogs usually packing and polluting the environment. Male investigates and lick anogenital region of female. Female exhibit lardosis and doesn‟t allow mounting until she is in proper heat. Methyl P-hydroxy benzoate, which stimulates the mounting reaction in male dog, was proposed as a pheromone released from vagina of bitch. Olfaction is main determinant communication between canine sexes

QUEEN:-

Cats are seasonally polyestrus. Shows estrus during late spring and early winter. Cats are induced ovulators. Estrus cycle lasts about 3 weeks. Proestrus lasts for 1-3 days some cats may not

show regular patterns of estrus cycle and anoestrus lasts for 90 days. Queens rub her head and neck against objects, rolling on the floor, refuses advances. Doesn‟t stand stationary and doesn‟t allow intermission by tom .In estrus estrous discharge is characterized by increased vacuolization for prolonged period. Rubbing head and neck against

38

objects and rolling on floor. Queen will elicit the attention of tom. Becomes on the chest and forelegs, rising of pelvis, elevation of tail are typical features she presents a perineal region which exposes a vulva. Minor vulval discharge may be observed but in general there no conspicuous changes in estrus. She allows grasping of neck and mounting. Tom licks anogenital region and is intensely attracted towards the queen Ton sniffs and grooms the female. Grips the neck of queen and mouth with vigorous pelvic thrust leading to intermission.

INDUCTION OF ESTRUS IN ANESTROUS ANIMALS Double injection schedule of PGF 2 alpha injection :- Anoestrous females are examined per-rectally for the exploration of genitalia. The females are injected with PGf 2 alpha injection intramuscularly. The onset of estrus is recorded. Cows show estrus within 72 hours. Those cows not showing estrus symptoms are given second dose of PGf 2 alpha injection 11 days after the first injection. Estrous is induced and fixed time breeding is done at 72 hours in those cows. Long term progesterone therapy :

Medroxy Progesterone Acetate (MAP) Melengesterol Acetate ( MGA) Short term progesterone therapy :

Norgestomet (Ear Implant), PRID, CIDR Exogenous P 4 acts like an artificial CL and inhibits estrus Withdrawal of P 4 induce estrus

NORGESTAMET/ SYNCHROMATE-B EAR IMPLANT :

A small hydron polymer implant impregnated with 6mg of Norgestomet is inserted s\c on the back of the ear of anoestrous animal. At the time of insertion 5mg of Oestradial valerate and 3mg of Norgestomet are injected intramuscularly (a single 2ml dose). The implant is removed after 9 days. AI done at Fixed time AI at 48 and 60 hours PROGESTERONE RELEASING INTRAVAGINAL DEVICE (PRID):

Stainless steel flat coil coated with an inert silicone rubber incorporating 1.55 g of progesterone Contains a 10 mg of Oestradial benzoate capsule. Left in situ (Vagina) of female for

39

12 days and then withdrawn. Estrus occurs 2-3 days after withdrawal. Fixed time AI at 48 and 72 hours or only at 56 hours after removal.

PROGESTERONE RELEASING

INTERNAL DRUG RELEASING DEVICE (CIDR)

INTRAVAGINAL

DEVICE

(PRID

CONTROLLED

T shaped silicone rubber, impregnated with progesterone and molded over a nylon spine.

A small nylon tail attached to the end of the CIDR protrudes from the vulva for easy removal.

Contains 1.9 g of progesterone.

Inserted in to the vagina of female using the applicator, left for 7 to 10 days and then withdrawn.

In cyclical animals a capsule containing 10 mg Oestradial benzoate is also inserted along with CIDR

Or prostaglandin can be given one day before withdrawal of insert.

Fixed time AI at 60 and 72 hours after removal of CIDR is recommended.

40

Handling And Management Of LN 2 Containers

Dr. S.C. Vora Veterinary College, Nagpur

Liquid nitrogen containers are very delicate by construction. Mishandling of LN 2 containers

due to lack of knowledge results in premature destruction of the containers. Any damage to the

LN 2 containers will reduce the efficiency of the container. This may result in increased evaporation

rate.

The liquid nitrogen container is double walled vessel. The inner chamber is suspended in

the outer chamber through neck tube, which is non-metal and bad conductor of the heat. This neck

tube prevents transfer of heat from outside to inside thereby preventing the rapid evaporation of

LN 2 .

Sudden moves and jerks vibrate the inner chamber. Thus side-to-side movement of the

inner chamber puts considerable stress on the neck tube and very often leads to mechanical

damage. The wall of the inner chamber is coated with high quality insulating material. Insulating

material is also filled in between the outer and inner chambers. Vacuum is created in between the

inner and the outer chambers. In the absence of the vacuum the LN 2 would be rapid evaporation of

LN 2 from the inner chamber.

A highly visible frost at the top of the LN2 container is the indicative of the rapid

evaporation.

Precautions during handling:

13. LN 2 containers should be kept in a cool place, direct exposure to Sunlight and hot air

should

be avoided.

13.

The room storing filled LN 2 containers should be well ventilated.

13.

Avoid direct contact of containers with hand floor.

13.

Avoid moisture on floor.

13.

Avoid injuries, drilling, puncturing, and scraping.

13.

Do not play with vacuum knob.

7. Use trolley for transport of LN 2 containers avoid rolling, pull or push with fiction on floor.

8. Do not put LN 2 containers one over other.

9. Always keep the lid over the container (except during putting in or taking out frozen semen.)

10. Do not interchange canisters from other LN 2 containers.

41

11.

Fill the liquid nitrogen slowly.

12. Make regular checks of the LN 2 level in the container. Any increase in the expected evaporation rate should be taken seriously.

13. Do not put undesirable material in the LN2 containers.

42

Infertility in Bovines

Dr. S. C. Vora Veterinary College, Nagpur.

Infertility: Infertility is a temporary state of reduction of reproductive ability of animals.

Forms of infertility: Infertility in large animals broadly classified in three groups.

1. Congenital forms of infertility.

3.Infections forms of infertility.

1. Congenital forms of infertility: -

2. Functional forms of infertility.

Congenital forms of infertility are associated with animals possessing anomalies of the

gonads and /or the reproductive tracts. Following are the some of the congenital causing infertility.

a. Free Martin: Free Martins have been noted in cattle, sheep, and goats but are more common in

cattle in concern infertility in females that have been born co twined with a normal male.

b. Ovarian hypoplasia: One or both ovaries may be underdeveloped, frequently observed

congenital malformation. Unilateral, ovarian hypoplasia results in reduced fertility. Whenever it is

bilateral, both ovaries are hypo plastic tubular part of the reproductive tract remains infantile.

Estrus cycle does not occur.

c. Segmental aplasia of Mullerian ducts: Congenital malformations of the uterus occur in all

species of domestic animals. The most frequently observed malformation is segmental aplasia of

the uterus it may affect any part of tubular genitalia.

2.Functional form of infertility: -

Anoestrus: Anoestrus when referred to in connection with physiologic estrus cycle is

usually characterized by quiescent functionless ovaries and reproductive tract.

Anoestrus in animals is classified as under:

a. True anoestrus.

b. Sub estrus/ silent heat.

c. Anoestrus due to persistent CL

a) True anoestrus: Both ovaries are small, quiescent and inactive. Uterus is small and flaccid.

Reason for failure of normal activity may be due to insufficient release or production of

Gonadotrophines to cause folliculogenesis.

It is never a primary condition but is conditioned by various environmental influences,

which probably through the mediation of anterior pituitary via hypothalamus depress initially the

43

ovarian activity. The main external influences are light nutrition and environmental temperature .The problem of shortage of feeds and fodders is well known in India. Treatment:

If the cause is due to chronic debilitating conditions this may be corrected, so the female

returns to good condition and near normal weight. Additional levels TDN and other nutritious diet needs to fed to this animals. Regular supply of green fodders and sufficient quantity of concentrates within regular deforming is necessary. Treatment by drugs and hormones:

1.Cocu tab.

: 2 tab per day for 30 days

2.Cofecu tab

: 1 tab for 20 days

3.Prajana cap. : 3 cap. For 2 days 4.Janova: 3 caps for 2 days

5. Estrona

6. Fertivet

(1% CuSo4, 125 ml is to be drenched before drug.) 7.Mineral mixture : 50 Gms. Per day for 30 days.

8. Inj. Receptal (GnRH injection)

9. Inj. Folligon (PMSG)

After 10-14 days 10.Progesterone treatment often associated with oestrogen has been Used to induce ovarian activity post-partum. 11. Estrogen both natural and synthetic have been used to treat Anoestrous. Hormonal therapy for the treatment of the anoestrus is useful in selected herds when used in healthy well-nourished animals. B) Anoestrus due to persistent corpus luteum:

It is the condition where CL persists without pregnancy and inhibits the normal cyclicity. Is also observed in pyometra, maceration and mummification. The persistent CL exerts an inhibitory effect on anterior pituitary.

: 2 cap per day for 3 days : 1 tab/day for 5 days

: 5 ml Intramuscular single dose. : 1500-3000 i.u. I/M may be repeated

Treatment:

.Inj. Dinofertin (PGE2a) is the best treatment.

: 25 mg I/m.

44

C) Silent estrus:

Reproductive organs are functional. Ovaries are undergoing cyclical changes but the external signs of estrus are exhibited. On rectal examination a high proportion of these will have a palpable CL, some may have a characteristic changes in the genital tract. Treatment:

1. Natural prostaglandin (Dinoprost or in synthetic analogue cloprostenol) is an important aid in

management of sub estrus in cows/buffaloes.

2. Dinofertin injection 25 mg/im.

Only effective between 6-16 days of estrus cycle. When uncertainly overage of CL, second dose of PGE2a should be injected after 11 days double insemination is practiced.

3. Clinical measures to improve detection of estrus e.g. Use of vasectomised bull.

INFECTITIOUS INFERTILITY Infertility in the animals due to infection or disease is common in India. Infertility due to some important infectious origin is as follows:

1) BRUCELLOSIS:

It is caused by gram ve coco bacillus Brucella abortus. This disease is important not only due to causing infertility problems in animals but also having zoonotic importance as causing undulant fever in man. Important symptoms of Brucellosis in herd is abortion, usually occurs from 6 th to 9 th month of pregnancy (gestation). Retained placenta, metritis with a subsequent period of genital discharge and infertility are the common squeal. For diagnosis of brucella: MRT/BRT test for herds screening is highly successful in indicating herds containing 1 or 2 brucella infected cows. For individual diagnosis and confirmation serological tests should be carried out. Now a days ELISA is also used for Brucella diagnosis. Treatment:

Because of the intracellular location of the organisms most of the antibiotics have no results. Symptomatic treatment should be carried out in aborting animals. For prevention: Hygienic measures are essential. Vaccination in calves 5 th to 7 th of age with strain- 19 vaccine may be used.

45

2) VIBRIOSIS:

It is caused by Compylobacter fetus venerealies spread at the time of coitus or at time of AI. Infertility or failure of conception lasting for a period of 2-6 months. Infertility is observed due to early death of embryo. Caused by C.foetus prolong irregular estrus cycle of 25-60 days are common following the first service of susceptible female to an infected bull. Treatment of individual cows by the I/U infusion of about 1 gm of Streptomycin together with Penicillin.

3) TRICHOMONIASIS:

Infertility, early abortion and pyometra are the main symptoms of Trichomonas fetus infection, protozoa responsible for economic losses in dairy herd.

INFERTILITY DUE TO NUTRITIONAL CAUSES Most causes of the reduced fertility or infertility are usually due to multiple deficiencies. In few cases only nutrients have a direct effect on fertility. Under feeding may be accompanied by poor quality of feed and deficiencies of protein, phosphorus & vitamin-A. Protein deficiency is usually accompanied by phosphorus deficiency and vitamin-A deficiency accompanied by protein and phosphorus deficiency. Young animals are more susceptible to nutritional deficiencies, which has consequent effect upon growth, puberty and sexual maturity.

A) Under feeding or starvation (inanition):

It delays on set of puberty and sexual maturity in heifers &may cause inhabitation of estrus cycle. In adult animals may cause failure of follicular development to maturity resulting in follicular Artesia. Underfeeding also delay in occurrence of post partum estrus in cows.

B)

Obesity:

Overfeeding causes obesity and may affect fertility in cattle. Obesity and sterility might occur due to hypothyroidism. Deposition of fat in the ovarian bursa and ovaries may interfere with normal ovulation and transport of ovum to the oviduct.

C) Protein deficiency:

underfeeding or where vitamin-A and

Quantitatively rarely observed except due to phosphorus deficiency occurs.

D) Carbohydrates deficiencies: -

It may cause inanition and loss of body weight and consequent effect on fertility.

46

E) Vitamins deficiencies: -

In cattle vitamin-A has considerable effect on fertility. Deficiency of vit-B is usually accompanied by reduced appetite.

F) Mineral deficiencies:

1. Phosphorus deficiency usually tends to occur when diet low in protein are fed and area where soil is deficient in phosphorus. Usual symptoms are delayed onset of puberty in heifers & failure of postpartum estrus in cows.

2. Copper deficiency :It causes delayed puberty, anoestrus sub estrus or poor pregnancy

results.

3. Cobalt deficiencies: It can occur in association with copper deficiencies results in failure

of estrus and delayed onset of puberty.

4. Iodine deficiency: Hypo/ Hyper thyroidism reduces the secretion of Gonadotrophines hormones by pituitary.

5. Iron deficiency: It results in anemia, debility, and lack of appetite and consequently

reduced intake of feed.

Treatment:

1. Mineral supplement: - Deltamin 50gm/day for 30 days.

2. Providing salt bricks.

3. Inj. of phosphorus.

47

Morbid uterus palpation and identification of different structures

Dr. Mukund Amle Veterinary College, Shirwal.

IDENTIFICATION OF DIFFERENT STRUCTURES IN FEMALE ANIMALS

A. RECTAL EXAMINATION OR PALPATION:

Rectal examination or palpation represents only practical domestic method permitting

direct examination of genital organs of cow, heifer of breeding age.

Equipments: -

Rubber or plastic sleeves with attached gloves is essential item not only for sanitary

reasons but also for protection. Record keeping almost as important as examination, it is therefore

advisable to palpate with hand that is not used for writing. Lubricant facilitates dilation of anal

sphincter and entry into rectum. Use of lubricant also reduces injury to rectal passage.

Restraint of Animal: -

In khoda (Travis) is advisable. First obstacle to entry into rectum is the anal sphincter so

pass the hand by making it cone shaped. Once dilated anus does not presents further difficulties.

Presence of hand into ampulla recti passage drawn out to elicit defecation reflex, which manifest

itself in peristalsis and tenesmus. Faeces must be explored with mucosa of rectum being only

structure between finger and organ to be examined. The removal of faeces from rectum is called as

backtrackings. If the hand is removed from rectum during the process of removing faecal matter

negative pressure is created on abdominal cavity &causes aspiration of air into rectum 7

consequent distention of intestine. Any attempt to examine through the wall of distended rectum

leads to trauma therefore should be avoided. Most common causes of rectal trauma are as follows

and elimination of them is highly helpful.

- To much force in manipulation.

- Manipulation in rectum distended by aspiration of air.

- Manipulation during peristaltic waves (tenesmus)

- Longer finger nails.

Pelvic or pubic frame provides an excellent hand mark for orientation during rectal examination of

genital tract.

Location & Examination of Cervix: -

It is located by first inserting arm just for enough to palpate pelvic inlet. Hand with finger is

rotated along one of the wall of pelvic cavity down to the floor over the side. Cervix is recognized

48

as a firm cylindrical nodular structure lying on middle line of pelvic floor. Examination of cervix for size, form and position whether it is open or close. The size of cervix depends on stage of reproduction and abnormalities. The size increases with number of parturition / abortion, cervicitis and age. The position of cervix normally located in pelvic cavity in cow. Suspension of cervix by broad ligament and indirectly by vagina permits considerable freedom for movement. This freedom for movement however is greatly dependant upon weight of uterine horn and body. With increase in weight of uterus it get fixed on pelvic brim and is relatively immovable eg. in pregnancy after 70 days, early puerperium, pyometra, mucometra, mummified fetus, ovarian/ uterine tumors etc. The common clinically apparent abnormalities of cervix are cervicitis, cervical and para cervical abscesses. Examination of non-pregnant Uterus / Retraction of Uterus: - Retraction or rolling of uterus is completed by grasping ventral intercornual ligament with middle finger & pulling uterus back to the pelvic cavity.

A ) Retraction by indirect method:

After the cervix is located and found freely movable it is pulled back as far as possible. Attempt is made to bring uterus up and close by moving retracted cervix upwards. Grasp the anterior broad ligament by keeping the thumb underneath the body of uterus turning hand outward lowering the bend finger and looking the broad ligament underneath. This broad ligament is grasped in angle between ovarian of horn and ovary. By sliding the fingers along the anterior edge will result in location of ovary. Median finger will lead to uterine horn. Horn is brought backwards and fingers are slide further medially until good hold or ventral intercornual ligament is obtained. This places the horn of uterus in posterior part of pelvic cavity & thus reaction is completed

B ) Retraction by direct method:

Retraction begins by grasping cervix and pulling it back as far as possible. The groove between two horns is then followed forward until anterior edge of dorsal intercornual ligament is

palpated. Then traction is completed as above. Then question to be answered by per-rectal examination of genital tract is whether pregnant or not pregnant. Physiological alteration of clinical significance involves primary size, symmetry, shape and tubularity. The tone of uterus during proestrus that is one to two days before estrus. Tones and excitability of myometrium undergo gradual but marked change which reaches its peak when cow or heifer shows sexual receptivity. Horns of uterus are turgid, swollen, coiled and also become

49

thicker contractibility of uterus diminishes. Uterus and luteal phase lacks tone and does not exhibit marked excitability. Symmetry of horn changes due to calving, pregnancy and abnormal contents. Abnormalities of uterus are freemartinism, white heifer disease, uterus unicornis, cervix duplex, chronic nonproductive metritis, pyometra, abscess, tumors of uterus (lymph nodes). Examination of Ovaries: - Average dimensions of ovary are; Length From pole to pole.

Width -

From surface to surface,

Height From attached border to free border Ovarian stroma with exception of corpus luteum and follicles is firm and nodular. Ovaries are found to hand width at about 10-12 cm laterally from midline, approx 1 or 2 fingers anterior to pelvic brim either at same level or bellow level of pelvic brim. Body of cervix and ovaries will lie in same line. To catch them move thumb and fingers alternatively from the base of horn to tip and then forward. Palpable functional structures of ovary:

Periodicity of estrus cycle depends upon or associated with periodic development of G.F. Ovulation is followed by development of C.L. (Yellow Body) and subsequent regression of CL accompanied by development of new follicle. This cycle is repeated unless and until fertilization takes place. Follicle on ovary is smooth, round, blister like, measures approx. 1 cm diameter to2 2.5 cm at peak of development. Fluctuation is felt under raised surface. Development of CL begins with ovulation. Ovulation depression on ovary is found around 12-24 hrs. after ovulation as field with blood clot referred as CL haemorrhagicum. In next 5-7 days proliferation and hypertrophy of luteal cells results in rapid development of CL more than 2 cm. in diameter. From 8-17 th day, the ovary contains fully developed CL, which go on regression further. Palpable abnormalities of ovary are hypoplasia of ovary, follicular cyst, luteal cyst, oophoritis and granulosa cell tumor.

Examination of oviduct and ovarian bursa : - Because of functional significance of oviduct detection of abnormalities is very important. Palpation of ovarian bursa involves locating mesoovarium, medial lateral to side of attachment of ovary. All finger then bend and slide underneath mesoovarium into ovarian bursa. Spreading fingers exposes bursa which is hooked in to thumb & lifted oviduct is then recognized on a cord like structure, this might be followed to infundibulum laterally & uterine horn medially. In normal

50

animal ovarian bursa should be free from adhesion. Abnormalities of adhesions only those attached, associated with enlargement of oviduct is detected clinically. Detectable abnormalities are segmental aplasia of oviduct of hydrosalpinx pyosalpinx.

B. PER VAGINAL EXAMINATION : - Vaginal examination is indicated only in certain selected cases. When indicated detailed

examine the external os of cervix, vaginal and vestibular mucosa & sub urethral diverticulum can be performed with vaginoscope or vaginal speculum. Types of speculum; 1.Glass speculum: Simple and economic

2. Plastic speculum : Easily sterilisable

3. Metal speculum: with or without looking arrangement (30, 40, 45cm).

Glass tube speculum is simplest and least expensive vaginoscpe it should be at least 4.5 cm long 2.5-3 cm in diameter and made from glass (3mm thick). Often animals are selected for vaginal examination after rectal examination. The external genitalia perineal region has to be thoroughly cleaned with disinfectant solution. Cotton mobbing is done. The speculum is lubricated with clear sterile lubricant and then inserted. The speculum must have to be directed upward in order to pass over ishiatic and then horizontal forward to have with handle of speculum in hand. The speculum jaws are closed while passing, Avoid entry in to suburethral diverticulum and urinary meatus open jaws of speculum and passage is dilated use hand like torch to observe interior of vagina. The normal mucosa appears pink moist. Slight congestion is seen on mucosa of entire genital tract at the time of estrus. Discharge of mucus from external os of cervix and its accumulation on vaginal floor is commonly seen while doing vaginal examination during estrus. Sign reflecting pathological conditions include discharge of purulent material from cervical canal, exudates fond on vaginal floor. Marked congestion, papules or pustules or ulcers are seen on vagina are seen on vagina and vestibule. Finding of tiny red elevation representing local proliferation of lymphatic tissue in vestibular mucosa. The elevations resembling granular vaginitis found primarily in the floor of vestibulum and are present in 50% of all females at breeding age. Granules do not reflect a disease interfering with fertility.

51

Evaluation of Semen

MACROSCOPIC CHARACTERS:

Dr. Mukund Amle Veterinary College, Shirwal.

The goal of semen evolution has been to predict fertility. The semen examination has great

diagnostic value in determining the cause, severity and degree of the pathological condition of

male genitalia. Naturally to have a high percentage of fertility it is essential that good quality

semen should be used for preservation and insemination. The main purpose of routine semen

examination is primarily to ensure the use of “normal” semen and further more the decision of

dilution rate.

Semen evaluation to be effective should be done with minimum delay after collection so

that the initial quality is not lost and the full potential is preserved. The samples showing

substandard quality should be discarded. For frozen semen the rejection rates are higher due to the

delicate processing methods.

Soon after the collection, till the evaluation and preliminary dilution are over, the semen

sample can be kept in water bath at 32 to 34 0 C. Fresh semen should not be stored for longer

periods prior to dilution (30 min. or more holding time increases the abnormal spermatozoa count.)

The neat semen sample should not expose to direct sunlight and cold storage (below 25 o C)

Volume, colour, consistency, density and P H of the semen sample are the tests commonly

used for preliminary assessment of the quality of the semen sample.

1. VOLUME AND GENERAL APPEARANCE: -

Volume of the semen measured immediately after collection in a graduated tube. The

semen should have a uniform, opaque appearance, free from hairs, dirt, dung, pus, urine and

lubricant material. Dung and dirt usually accumulates at bottom of the tube while the lubricant

material usually remains at the top of the semen. Good quality semen usually has foam at the top,

immediately after the collection.

Low volume is not harmful, but if accompanied by low concentration, limits the number of

the doses produced. The volume of buffalo bulls is comparatively less than cow bulls and in the

same age group.

Factors affecting the volume of semen;

i. Age :- Younger bull produce low volume of the semen, which increases up to 6 years of age in

cow bull and up to 7 years of age in buffalo bulls.

52

ii. Size of the male : Volume is related to size and body weight of the bull and may be hereditary

trait also.

iii. Precollection sexual stimulation : Adequate sexual stimulation through false mounting and

proper restraint increase the volume of the semen .

iv. Frequency of semen collection :

First two ejaculates are nearly of equal volume and thereafter volume decreases. However, if the

interval of collection is more than 7 days, second ejaculation may yield comparatively higher

volume. Usually, volume remains nearly constant if frequency of collection is 3-7 days.

Volume reduces if collection is taken more frequently.

The semen volumes in different species are;

Sr.

 

Volume of the Semen / ejaculate

Sr.

 

Volume of the Semen / ejaculate

No.

Species

No.

Species

 

1 Cow Bull

0.5

-

17 ( 4 ml )

 

4 Boar

150 200 ml

2 Buffalo Bull

2.5

-

8.2 ( 3 ml)

5 Ram/ Buck

0.8 1.2 ( 1 ml)

3 Stallion

60 100 ml

6 Dog

2 19 ( 7 ml)

2. COLOUR : -

Colour of the semen is influenced by its consistency. Semen with watery consistency is

readily distinguished from that with a viscid consistency. The normal Colour of the semen is

grayish white to yellowish white. Creamy samples are usually observed in cow bulls, rams and

bucks, while they are usually milky white in buffalo bulls. Any abnormal colour may be indicating

pathological condition in the genital tract. Ejaculates with abnormal colour are discarded.

Varying shades of yellow colour are considered to be due to riboflavin content (which may

be inherited in some bulls) however it may be also indicate presence of pus or urine in the semen.

Reddish or pinkish colour indicates presence of blood and may be due to injury in the lower

part of the genital tract. Pinkish colour is seen due to degenerative changes

A grayish or brownish tinge to the semen may be due to presence of decomposed blood in it

and indicate an old injury in the upper part of genital tract. In haemospermia, where there is a

tumor or cold abscess on any part of reproductive tract gives brownish semen.

A greenish color may indicate purulent degeneration suggestive of pathological condition

of genitalia.

53

2.

CONSISTENCY : -

The consistency or viscosity of semen gives a rough indication of the concentration of

spermatozoa. The normal bull semen may be creamy or milky in consistency depending on the

concentration of sperm. High quality semen should be thick, cloudy and should show swirls.

Cloudiness indicates vigorous activity of the spermatozoa. In certain pathological conditions of

testis, epididymis and accessory glands the consistency of semen becomes milky to watery.

Consistency greatly influences the colour of the semen. The approximate relation of colour,

consistency and the concentration of spermatozoa are as follows.

Colour

Consistency

No. of Sperms (millions / ml)

Quality

Creamy

Thick

Over 2000 1500 2000

Excellent

Thin

Very good

Milky

Thick

1000- 1500

Good

Thin

500- 1000

Fair

Cloudy

 

100- 500

Poor

Watery

 

Below 100

Bad

4. DENSITY : -

It gives the rough estimation of the concentration of spermatozoa in the semen sample. It is

done holding the semen collection tube against natural light. The density gradation with

approximate concentration of spermatozoa is as follows,

Density

Sperm Concentration

Density

Sperm Concentration

D

Up to 400 millions/ml

   

DD

400

to 800 millions/ml

D (D)

600 millions/ml

DDD

800

to1200 millions/ml

DD (D)

1000

millions/ml

DDDD

Above 1200 millions/ml

DDD (D)

1100

millions/ml

5. HYDROGEN ION CONCENTRATION (pH):-

pH is estimated by 3 methods.

i. B.D.H. Capillator pH indicators Bromothymol blue (pH range 6.0 7.6 ) and Bromocresol

purple (pH range 5.0 6.8 ) are used for this purpose. Equal quantity of semen and indicator

are taken in a watch glass. After thorough mixing, the mixture is taken in a capillary tube and

the color is matched with the colors of B.D.H.Capillator. The corresponding figure indicates

the pH of the semen.

54

ii. Indicator paper :- A strip of the indicator paper is torn from the indicator book and the tip is dipped in a drop of semen. The change of color is seen and is compared with color on indicator book. The corresponding figure indicates pH of the semen. Paper No. 5075 pH range 5.0 7.5 & Paper No. 6590 pH range 6.5 9.0 iii. pH Meter : - The best and the most accurate method. The P H of semen in various species is as follows:

Species

Acidic- Neutral pH

Species

Neutral to Alkaline pH

Cow Bull

6.4

6.8

Stallion

7.3

7.9

Buffalo Bull

6.2

6.6

Boar

7.3

7.9

Ram

5.9

6.8

Cat

 

Buck

5.9

6.8

Cock

7.02 7.18

Dog

4.9

6.9

   

The pH 7.0 and above of bull semen is observed in bull extensively used, incomplete ejaculation, inflammatory / pathological condition testis, epididymis and seminal vesicle. Highly motile semen is acidic where as poor motile and low concentration of sperm leads to alkaline semen. MICROSCOPIC CHARACTERS 1. MASS ACTIVITY: Motility or movement of spermatozoa is an index of their activity and it represents the quality of the semen. The live sperms are motile but all the immotile sperms are not dead. The epididymal spermatozoa are motionless, but during the course of ejaculation, after coming in contact with the accessory gland secretions, they gain vigorous motility. The spermatozoa motility is judged as gross or mass activity and as individual motility. Mass / Gross Activity:

Mass activity of the semen is examined immediately after collection. The collection tube containing the semen sample is closed with an aluminum foil or cotton plug and is kept in a water bath at around 30- 35 0 c temperature. A drop of fresh undiluted semen is placed on a clean pre warmed (37 0 C) slide without applying cover slip and observed under the lower power of microscope. Hanging drop preparation is most satisfactory but more difficult to prepare. Good samples show a series of rapidly changing swirls, waves and currents of sperm motion. On the basis of the mass movement of the spermatozoa, the semen is graded into the following 5 categories.

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Movement of sperm

Grades

No mass activity.

0 Nil

 

Movements slightly vigorous but no eddies or currents

formed

+ Poor

 

Wave formation with slight whirls which moves slowly across the field

+ +

Fair

Rapid and vigorous waves with whirls/eddies which change with rapidity

+ + +

Good

Extremely rapid movements and churning of swirls and eddies

+ + + +

V. Good

The type of movement is influenced by the size and depth of the drop examined, temperature

of the semen sample, rate of the sperms and sperm concentration. Semen samples with mass

activity grade at least + + + are accepted for A. I. purpose.

2. INITIAL MOTILITY:-

This gives rough idea of the % of motile sperms and their vigor. Four types of sperm

movements are observed.

Progressive Sperm moves fast in forward direction.

Circular Movements restricted to a radius of approximately the length of the sperm.

Oscillatory Side to side movements without change of place. ( in aged semen samples)

Reverse Observe very rarely.(Circular / reverse movements are usually signs of cold

shock)

Semen is diluted (1:100 or more) in an isotonic medium viz. normal saline, 3 % sodium

citrate dihydrate solution or Ringers bicarbonate solution. A drop of this dilution semen is placed

on a clean, pre-warmed slide (37 0 C) and examined under low as well as high power of microscope,

using cover slip.

Semen samples with minimum motility score of + 3 are selected for further processing.

Semen samples with initially less than 60% live spermatozoa reduce fertility and are not good

preservation. Semen samples with initially less than 50% live spermatozoa have doubtful fertility.

The motility scoring is done as follows;

S

 

Motility

Approx.

 

N

Movements of Spermatozoa

Score

sperm

motility

Grade

1

Nil / No movements

 

0

0 %

Nil

2

Mostly weak and oscillatory.

+

1

20

%

Poor

3

Motile sperm with rapid vigorous movements.

+

2

40

%

Fair

4

Very rapidly, vigorous and progressive movement.

+

3

60

%

Good

5

Most vigorous with progressive swirling activity.

+

4

80

%

Very Good

6

Highly vigorous, progressive waves in all directions.

+

5

100 %

Excellent

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Factors affecting sperm motility: -

Frequency of semen collection: - If the collection interval is more than one week, the motility is better in second ejaculate. However, if collection interval is 3 -7 days, no difference in motility between first & second ejaculate is noted. If collection is done on successive days for a long period, the motility is greatly reduced.

Season: - Spring season produce more motile semen than summer and winter. Hot and humid climate is harmful for production of high motile sperms.

Age: - Young bulls produce comparatively less motile semen. Motility increases with age of bull. Sudden drop in the motility of good quality semen may be due to faulty collection, faulty handling of semen, and contamination or disease conditions like testicular degeneration. It has been observed that some of the non-motile semen sample may gain progressive motility on treatment with diluting media. (Especially in case of buffalo semen). Also some times though the spermatozoa are alive, they are immotile initially. 3. ESTIMATION OF TOTAL CONCENTRATION: - Macroscopic examinations like consistency and density of the semen samples provide a rough estimate of the semen concentration. However, to provide optimum no. of live and motile

spermatozoa per dose of liquid or frozen semen, it is essential to estimate the actual concentration of the spermatozoa in the semen samples. By knowing the initial concentration of the spermatozoa, the dilution rate of the semen can be decided. It is generally accepted that at least 10-12 millions motile and live sperms must be deposited in the cervix to obtain the optimum conception rate. For achieving this, 10-15 millions and 20-25 millions spermatozoa are provided per dose of liquid or frozen semen respectively. Methods of estimation of sperm concentration: Opacity tube method, Photo-electric colorimeter method and Direct count by use of Haemocytometer

a. Photo-electric colorimeter method:- This instrument measures the percentage of light

transmitted through a light-absorbing medium. The percentage of transmission is a function of the concentration of the light-absorbing agent in the medium. The same principle is applied to estimate the sperm concentration in the semen sample. Data in a tabular form is used to correlate the

percentage of transmission (meter reading) to corresponding sperm concentration. b. Opacity tube method: - Density of the semen sample is compared with Barium sulphate or other density standards.

57

c.

Haemocytometer method: - Following solutions are required

i. Normal saline

Diluting fluid, ii. Diluting fluid is prepared as-

Sodium citrate dihydrate

3.0 gm

Eosin yellow

100 mg

Formalin (commercial)

1 ml

Distilled water

100 ml

Dilution of semen sample: - A dilution of 1:1000 is used for estimation of sperm

concentration. Using an ordinary 1ml glass pipette 0.1ml of semen sample is taken. It is transferred into a test tube in which 9.9 ml of normal saline is added. This gives a 10 ml solution with 1:100

dilutions. 1ml of this solution is added to 9 ml of diluting fluid to get a 10 ml solution with a dilution of 1:1000.

Charging of haemocytometer:- The cover slip is fixed

so that the colours of spectrum (Newton‟s rings) are seen at the interphase. After through mixing, a representative drop of (1:1000) diluted semen sample is put at the edge of the cover slip with the help of Pasteur pipette, so that the fluid covers the entire chamber.

Counting of spermatozoa: The Neubauer‟s chamber is divided into 9 large primary squares,

each having an area of 1sq.mm. All these 9 squares are enclosed from all sides by treble lines. The 4 corner squares are again divided into 16 small squares each. The height of the chamber (depth) formed after fixing the cover slip is 0.1mm. The spermatozoa in 1sq.mm.are to be counted (i.e. spermatozoa in 16 small squares) for this Count the spermatozoa in 16 small squares of all the 4 large corners squares (i.e. total 64 small squares) and take average by dividing 4.

over the Neubauer‟s chamber slide

Calculations: „N‟ no. Of sperms are present in 16 small squares. Volume of fluid containing „N‟ sperms = Area (L x W) x Height = 1mm x 1mm x 0.1mm

„N‟ No. of sperm in 10 N

= 0.1mm 3 = 1mm 3 (1 ml = 1000 mm 3 )

i.e. 1m. fluid contains 1000 x 10 „N‟ spermatozoa. This fluid has 1:1000 dilution of semen. Hence1ml of original (neat) semen contains = 1000 x 1000 x 10 „N‟ spermatozoa = 10 „N‟ x 10 6 (million) spermatozoa.

58

Note : To avoid double counting of a sperm or omission of sperm, a procedure is followed in counting. Those sperms lying on the top line and left hand line are included in that square while those lying on bottom line and right hand line are excluded from that square. Factors affecting sperm concentration:

a. Sexual excitement: Sexual excitement caused by restraint and false mount increases the sperm concentration.

b. Frequency of collection: No variation in sperm concentration is observed when the

frequency of collection is 3-7 days. However concentration reduces when frequency is at 24-48 hrs intervals. When collection interval is more than 9 days, 2 nd ejaculate more concentration than 1 st . When collection interval is 3-7 days no difference between 1 st and 2 nd ejaculate is observed.

c.

Season: has no significant effect on sperm concentration in cow bulls as well as buffalo

bulls.

d.

Age: has no significant effect on sperm concentration, however, total output of spermatozoa

per ejaculate is increased in adult bulls.

e. Individual variation: Due to difference in genetic potential, the concentration varies.

f. Other factors: A sudden drop of sperm concentration can be attributed to overuse or

disease, if not, premature ejaculation or faculty collection. Disease conditions reducing the sperm concentration cause of motility besides increase in abnormal sperms.)

3. LIVE AND DEAD COUNT OF SPERMATOZOA:

A high percentage of live and progressively motile vigorous sperms are essential for good quality semen with high fertilizing capacity. The motility tests of semen (viz mass activity and initial motility) have their limitations in estimating the quality of semen. A fairly high percentage of dead spermatozoa may not be apparent in motility examinations because of the inactive sperms are moved by the movements of live spermatozoa. Hence to substantiate/support the findings of the motility tests it is essential to estimate the percentage of live and active spermatozoa. Eosin-Nigrosin stain (Hancock, 1951) is used for staining of semen smears. Use of separate solutions of Eosin yellow 5% and Nigrosin 10% Or Combined stain:- Contains eosin yellow 1.67 gm and Nigrosin 10gms in 100ml distilled water .The stain is prepared by dissolving the ingredients in distilled water in a hot water bath finally the solution is filtered through The staining mixture should be isotonic with the semen and its pH should range from 6.8-7.Temperature at which the semen and stain are mixed and the duration of staining is very important factors. The

59

stain solution and the semen sample should be maintained at same temperature (optimum temp. range :- 30-35 0 c) in a water bath prior to mixing. The drop of semen is taken in a watch glass, to which 2-4 drops of combined stain solution / 2 drops of separate stains are added. A time limit of about 2 minute is sufficient for proper mixing of stain and semen sample. The smears are prepared by routine procedure and are air-dried. Spermatozoa are observed under high power lens of microscope. The dead spermatozoa take eosin stain and appear pink in colour while the live spermatozoa remain white. Nigrosin provides uniform dark (purple) background. Counting of spermatozoa: partially stained (more than half) spermatozoa are considered dead. In all 333 sperms from different fields are counted by hand tally counter and the dead spermatozoa are noted separately on the paper.

% of dead spermatozoa = No. of dead spermatozoa (N) 3/10 or 2 to 4 smears are prepared using different sub samples of the same original sample from each smear 100 spermatozoa are counted by hand tally counter and finally avg. is calculated to find out the dead spermatozoa.

Factors affecting live and dead % of spermatozoa:

a. Frequency of semen collection: If the collection interval is more than a week, the second

ejaculate may have more % of live spermatozoa. If the collection interval is once in two days to once a week, the %may not markedly vary in1 st & 2 nd ejaculate. In successive ejaculates, the % declines after 2 nd ejaculate.

b. Age of male: may not have more effect. In general, bulls over 3 years age produce higher %of

live spermatozoa. c. Season: lower live % is noted in hot humid season.

5. ABNORMAL COUNT OF SPERMATOZOA: - First attention to spermatozoal aberrations was paid by Williams, (1920). It was further emphasized as a valuable diagnostic aid in assessing the potential fertility of bulls by Williams and Savage (1925,1927) and Lagerloff (1934). The male fertility depends upon morphologically normal spermatozoa present in ejaculated semen. A maximum of 15-20% of total abnormalities of sperms are does not affect much fertility of male animals, however above 30 % total abnormalities are affect fertility. Most of workers agree

60

that semen of fertile bull should not have more than abnormalities of head 4%, mid piece 8 %, tail 2 % and free heads 6 % and total 20 %. Factors affecting abnormal count of spermatozoa :

Several factors affect the abnormal count of the spermatozoa viz. defective spermatogenesis, systemic diseases, inadequate nutrition, adverse environmental conditions, handling, smearing and staining procedure, frequency of semen collection, age and genetic factors which may increase the % of abnormalities. The first two rapid ejaculates generally do not vary the % of abnormal spermatozoa but further collections increase the incidence of abnormality. It has been observed that the frequency of collection at 48 to 72 hrs from zebu bulls lowers the abnormality. Malmberg (1965) observed a decrease in the proportion of abnormal sperms with the increase in the age of the bull. Staining and preparation of slides:

Eosin Nigrosin combines stain (Hancock, 1951) can be conveniently used for abnormal count of sperms. The spermatozoa are observed under either high power or oil immersion lens of microscope and total of 100 spermatozoa are counted from each slides using various fields. (Making thin smears on clean slides helps to reduce the artifacts). Classification of abnormal spermatozoa:

I. Classification suggested by Lagerloff (1944) A) Pathological forms:

a) Head: Mirocephalic, Macrocephalic, Elongated, Narrow head Pear shaped, Round, Giant, small

free head, deformed head.

b) Midpiece: - Abaxial, Swollen, Filiform, Coiled, Bent, Double.

c) Tail: - Coiled, Bent, Short, Double.

d) Underdeveloped spermatozoa: -head mostly rounded with tail coiled around the head.

Immature forms

e) Proximal protoplasmic droplet: - bend on midpiece close to the head.

f) Distal protoplasmic droplet: - bend on midpiece close to the main piece of tail. II. Classification suggested by Blom (1948), Bishop etal (1951) a. Primary abnormalities (Defective spermatogenesis due to disorders in seminiferous tubules or germinal epithelium):- Pyriform head ,tapering head, narrow head, dwarf head , giant head, short thick head , other abnormal shaped heads, loose abnormal heads, loose abnormal head, normal

61

head with highly coiled tails, double forms of head / tail /

midpeice.

b. Secondary abnormalities (due to degenerative changes occur in their passage through efferent

ducts, epididymis and vas deference.): - Detached normal head, Proximal and distal protoplasmic

droplet, bent tail, and detached galea capitis.

c. Tertiary abnormalities (are due to faulty handling of semen, excessive agitation, over heating. cold shocks, erroneous methods of slide preparing, staining etc.) eg. Loosely coiled midpiece or tail.

II. Recent classification suggested by Blom (1971)

A. Major sperm defects: - Underdeveloped sperm, double forms, acrosome defect (knobbed

sperm), decapitated sperm defect, diadem defect, pear shaped head, head narrow at the base, head with abnormal contour, small abnormal head, free pathological head, corkscrew defect, other middle piece defects (including tail stump), proximal protoplasmic droplet, pseudo droplet, dog

middle piece, Abaxial attachment of

defect.

B.

Minor sperm defects:- Narrow heads, small normal heads, giant & short broad heads,

normal free head, detached acrosomal cap, abaxial attachment, distal protoplasmic droplet, simple

bent or coiled tail, terminally coiled tail. IV. Certain morphological sperm defects of hereditary origin.

A. Defects of sperm Head:-

1. Decapitated sperm defect:- (disintegration of sperm) :- virtual absence of intact sperm in

semen. Ejaculate contains only free heads & tails. Sexual behavior & clinical examination of genitalia do not show any abnormality. Seen in jersey bull.

2. Knobbed sperm (acrosome) defect: - Sperm count and motility normal but 90% sperm show

abnormal acrosome. Common in HF also noted in boars.

3. Diadem defect: - (pouch formation of nuclear envelope):- Pouch is formed by invagination

of nuclear membrane. It appears as sparkling, round or elongated white sport in the head. These structures give impression that a neck less has been tied around neck of spermatozoa. It is associate with disturbance in spermatogenesis. Noted in bovines & porcine.

A. Defects of midpiece: -

1. Corkscrew sperm defect: - Characterized by an irregular distribution of mitochondrial

sheath. The incidence of proximal protoplasmic droplets is high observed in aged bulls & in associated with testicular degeneration in some cases observed in young bulls also.

62

2.

Psuedo droplet defect: - Local thickening somewhere along midpiece. Initial motility, poor

concentration & genitalia normal.

B. Tail defects: -

1. Dag defect. Strongly coiled tail

Other cells found in semen: - 1.Medusa formations: these are portions of from ciliated epithelium from the efferent ducts of testis and have brush like projection. These cells are seen in great number in sever testicular hypoplasia.

1. Primordial spermiogenic cells

2. Spermatid / spermocytes

3. Free floating protoplasmic droplet

4. Giant multinucleated cells seen in testicular hypoplasia or degeneration.

5. Preputial cells

6. Leucocytes because of infection in genital tract,

7. Erythrocytes are the other types of cells found in semen.

8. Degenerating sperm cell cluster seen in testicular degeneration.

63

Record keeping in Artificial Insemination

Dr. S. C. Vora Veterinary College, Nagpur.

Records is the only sensible basis for taking decisions about alterations in policy (such as

changes in stoking rate or the introduction of new animals) the only way of ascertaining whether

the targets of reproductive.

Information comes from records through their proper collection and critical analysis the

record should include the following elements.

1. Individual Card /record (Female record)

For each animal on which life events of the animals can be listed in chronological order.

The information stored on cards would include:

a) Name /identity (ID):

b) Date of birth:

c) Location number:

d) Calving date for each location :

e) Problems at calving:

f) Retention of fetal membranes:

g) Any vulval discharge:

h) Date of estrus:

i) Service date:

j) Pregnancy diagnosis (PD):

k) Veterinary treatments & dates:

l) Drying off dates:

m) Dates culled with reasons:

A) Semen record: