Development of Technologies

The institute has developed several technologies since its inception that were transferred to the farmers to increase the production and reproductive efficiency of their buffaloes. During the period under report 55 lay man inseminators were trained in artificial insemination and animal husbandry in 3 training programs. Many of the farmers trained in this institute are achieving ~60 % conception rates with the frozen semen from this institute. The developed technologies are also transferred through field visits, kiasn melas, radio and TV talks and web portal based extension activities. Books, bulletins and popular articles are regularly written by scientists for dissemination of knowledge of scientific buffalo husbandry to the farmers. Some of the technologies which found acceptance with users are presented below.

Production, maintenance and dissimination of superior germplasm.

Institute maintains a high pedigreed herd of Murrah and Nili-Ravi buffaloes. The institute has been undertaking breed improvement programme through selective breeding since its inception. The genetic potential of bulls is evaluated through progeny testing. Due to intense selection pressure, production performance of Murrah and Nili-Ravi herds improved from about 5 kg in 1991 to 8.25 in Nili Ravi and 8.0 kg in Murrah during 2013-14.More than two lakh doses of frozen semen from test bulls and over fifty thousand doses from progeny tested bulls are available for Murrah breed improvement.About 454 Murrah and 302 Nili Ravi bulls of high genetic merit have been supplied to various developmental agencies and village panchayats in 12 States for increasing milk production through genetic improvement.Under field progeny testing program in adopted villages, 17623 AIs were done so far with frozen semen of test bulls with conception rate of 48%.

Improved protocol for buffalo semen cryopreservation

A simple, reliable and economical method for freezing of buffalo semen has been developed and found to be effective to freeze the static ejaculates successfully, a phenomenon specific to buffaloes which greatly reduces the efficiency of utilization of buffalo semen for artificial insemination. A large proportion of buffalo semen ejaculates collected during summer months are rejected due to the high incidence of post-thaw backward motility of sperm cells. Through thorough investigations about the phenomenon, stage ofglycerolization was identified to be the most critical stepresponsible for backward sperm motility. Glycerolization at room temperature during initial stage of semen dilutionreduced/eliminated the backward motility due to which 20percent more ejaculates could be preserved annually,thereby enhancing the frozen semen production.Overall semen freezing protocols improved resulting in almost 15% point improvement in post-thaw motility and improved frozen semen quality and fertility on artificial insemination.

Area-specific mineral mixture

Surveys of feeding practices carried out in Haryana revealed deficiencies of essential minerals like calcium, phosphorus, zinc and manganese in 70 percent of buffaloes. On the basis of analysis of mineral intake vs requirement an area specific mineral mixture was developed. Seventy per cent of the buffaloes suffering from anaestrus conceived within a period of 2-4 weeks of feeding the area specific mineral mixture. The mineral mixture improves feed intake, milk production and reproductive efficiency. Institute has been preparing and selling mineral mixture to the farmers at no profit no loss basis.

Feeding standards for different categories of buffaloes

Feeding standards have been developed for different categories of buffaloes, viz. growing males, growing heifers, lactating buffaloes and pregnant buffaloes. Nutrient requirement for heat and humidity stress was also estimated and published.

Ultrasonography for fetal age and sex determination, pregancy diagnosis and monitoring ovarian activity

Ultrasonography guided fetal age and sex determination technology has been standardized. The accurate diagnosis can be made at 55 day of gestation in buffaloes in contrast to 50 days reported in cows. By ultrasonography fetal age can be accurately assessed that is useful in better management of pregnant buffalo at the time of calving. The length of gestation in buffalo can be estimated by following standard chart that is developed for crown-rump length of buffalo fetus on different days postinsemination. When this plot was used for determining the age of fetus in pregnant buffaloes the exact date of mating/gestation could be predicted with a precision of +- 1 to 2 days during first 60 days of gestation. The non-invasive technique of ultrasonographic scanning has been standardized for diagnosis of ovarian activity. This technique is very useful for follicular dynamics studies. With the use of this technique, time of ovulation can be predicted very precisely to allow fixed time insemination.A protocol has been standardized for establishment of early pregnancy diagnosis in buffaloes. With ultrasonic scanning, pregnancy could be diagnosed as early as 26 days post insemination. The technique can be used to assess date of service in case of unobserved mating.

Identification of molecular markers for MAS

RAPDs, Microsatellites and traits governing specific genes as growth hormone, seminal fluid protein gene specific primers based buffalo genome characterization done for identification of genetic diversity and markers for higher milk profuction and bull performance Buffalo genomic study has just been initiated in collaboration with NBAGR. A twenty nucleotide base pair length having di-nucleotide repeats have been identified showing polymorphic expression of milk production in low and high milk producing buffaloes. Study revealed more than 30 percent dissimilarity between high and low yielding buffalo genotypes.

OvSynch plus protocol for estrus induction in buffaloes

Anestrus, in pubertal heifers and postpartum buffaloes, is the primary cause for low reproductive and productive performance of buffaloes. The condition is associated with the presence of static ovaries and though follicular development may occur, none of the ovarian follicles becomes mature enough to ovulate. In anestrus animals, doninant follicle (DF) undergoes atresia instead of ovulation. Analysis of ovarian response of anestrus buffaloes to ‘Ovsynch’ protocol revealed that only the buffaloes with a large DF (>9mm) at the time of first GnRH injection respond well to this treatment. However, such an accurate assessment of follicular size is difficult under field conditions with routine per-rectal palpation. Hence, to ensure consistently similar ovarian follicular picture of all anestrus buffaloes at the time of first GnRH injection, a new protocol was developed and named ‘Ovsynch Plus.’ In this protocol, an injection of PMSG is administered 72 h prior to the first GnRH injection of Ovsynch treatment, in order to support ovarian follicular development so that at least one large follicle is available 72 h later for responding to the GnRH injection with ovulation/ luteinization. Resulting luteal structure in the ovary is then subjected to luteolysis by PGF given 7 days later. Further administration of GnRH ensures synchronous ovulations of preovulatory follicles to allow fixed time insemination of treated animals.
Developed OvSynch protocol for estrus induction in buffaloes.The major advantage of this protocol is that it induces oestrus in cyclic as well as acyclic animals within a close window. Buffaloes not coming into estrus within the defined period following this protocol also become cyclic and get pregnant within one month of treatment, if initiated during breeding season.

Embryo transfer technology

Efforts have been made in developing and improving the embryo transfer technology for buffaloes which has resulted in the production of 20 calves at this Institute. Technology for large scale production of in-vitro matured and in-vitro fertilized embryos using slaughter house ovaries has also been developed. The embryo cryoprservation technique has been standardized. This technique has been standardized for in-vitro maturation of oocytes obtained from abattoir ovaries followed by their in-vtiro fertilization and culture of the resulting embryos to transferable stage. The technique of IVF will be of immense use for faster multiplication of elite germplasm and progeny testing of bulls after collecting oocytes from live animals.

Scrotal circumference for bull selection

Scrotal circumference of Murrah buffalo males is highly correlated with age and body weight and it can, therefore, be used for pre-selection of breeding bulls at an early age. For mature (>600 Kg BW) Murrah buffalo bulls (n=86), mean SC values were 35.23 cm, with S.D. of 3.00. Therefore Murrah bulls having scrotal circumference 29 cm (Mean -2 S.D.) must be excluded from the breeding programme, while males with SC of over 41 cm (Mean +2 S.D.) should qualify as the best semen donors.Scrotal circumference is highly correlates with semen production potential of a bull.

Induction of lactation

Farmers rear the dairy animals for milk production and livelihood but they are commonly facing the problems of conception failure, long calving interval, anestrous, cystic ovaries, specific abortions and repeat breeding. They can benefit by inducing such animals into lactation by induced lactation therapy. The buffalo is weighed and appropriate dose of hormones, Estradiol- 17b and progesterone @ 0.1 mg/kg body weight/day each, is calculated for seven days therapy, dissolved in absolute ethanol and stored. On the day of treatment, 1 ml of each hormone solution is administered subcutaneously in the morning and evening at an interval of 12 hours, for seven consecutive days. Thereafter, on day 17,19 and 21 of treatment, 10 ml Largectil injection and on day 16, 18 and 20, injection of 20 mg of Dexamethasone are also given intramuscularly. Between 15th and 21st day of treatment, udder massage is given for fifteen minutes each in the morning and evening daily till the udder is turgid with milk, which is usually around 21st day when milking is started. The milk becomes normal in physical and chemical properties within 10 -15 days of start of milking and the amount of milk yield increases with time. Almost 60-75 percent of the buffalo’s milk yield potential can be achieved following induced lactation.

Colostrum for higher growth and calf survival

Higher levels of immunoglobulins absorbed within 16 h of birth, reduce the mortality in calves and result in faster growth rate by 20-22 percent. High titre of circulating immunoglobulins in calves at an early age of 24 h showed the association with weight gain upto the age of 2 years. Status of immunoglobulin levels at such an early age could also predict the health status of calves. A critical level of these blood proteins required for the survival of calves has been assessed.

Antioxidants in survival and growth of neonates

Advanced pregnant (270 to 280 days’ gestation), buffaloes are administered two doses of antioxidant micronutrients, consisting of vit A (Palmitate), vit D and vit E (dl- alpha 3 Tocopherol acetate, within 30 days before calving, at 15 days intervals. These buffaloes secreted 25-80% more Ig protein in colostrum than control buffaloes. Calves born to treated buffaloes were also supplemented with mineral mixture @ 5 g/calf/day, colostrum feeding @ 10% of birth weight, concentrate mixture started 10 to 15 days after birth and green folder offered after 3 weeks, in order to achieve high growth rate and survival. Calves born to vitamins administered buffaloes and further supplemented with mineral mixture gained 10 percent higher body weight and 30% better immunity status. Calves bearing higher body weight and better immunity are economically more rewarding for meat and milk industry.

Uromol preparation

Uromol is a compound prepared by heating urea and molasses in the ratio of 1 : 3 and then mixing it with equal amount of wheat bran/deoiled rice bran. Four kg urea along with 12 kg molasses is slowly heated in a container for 30 minutes. Then equal amount (16 kg) of wheat bran or deoiled rice bran is mixed in it and the mixture is cooled to room temperature. This material contains 36 percent DCP and 72 percent TDN and can replace conventional compound feeds in the ration of buffaloes yielding 8-10 litres milk/day.

Urea molasses mineral blocks (UMMB)

Urea molasses mineral blocks are prepared in the same way as Uromol, except with the addition of mineral mixture, salt and binder. By ad-lib feeding these blocks along with other feed ingredients, about 20 percent of the conventional concentrate mixture can be saved. UMMB prepared by the ‘cold process’ technology has yielded even better results.

Superior isolates of anaerobic fungus

Superior isolates of anaerobic fungus were isolated and evaluated for ability to increase in vitro digestibility of straw by buffalo rumen microflora. Such isolates have the potential to be used as feed additives.

Enzyme supplementation

Fibrolytic enzyme supplementation can be used as feed ingredient in the concentrate mixture of calves to increase the growth rate. Further, the cost of enzyme can be reduced by using feed grade enzyme or enzymes used in textile industry (cellulase) and paper industry (Xylanase).

Thermal stress management

Microclimate modifications with supplementation of niacin @ 6 gms/day/animal, yeast @10 gms/day/animal and mustard oil @150 gms/day/animal; enhance milk production of lactating buffaloes by reducing thermal stress.

DNA bank

DNA repository of about 3119 buffaloes has been established at the institute for genome analysis. Phenotypic data on all the animal is being collected which shall be used for establishing linkages with performance traits and identification of molecular markers.

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