Reproductive Health Management in Ruminants

by : G.M. Wani



Abstract
Reproductive health management in Ruminants is a buzz word called Total Quality Management (TQM). Goals to achieve reproductive efficiency means 12-13 months calving interval, low repeat breeding and early rebreeding after parturition. Less than 2% abortion & less than 10% retained placenta and metritis in in herds may ensure less that 18% culling rates.
In a developed country like USA reproductive efficiency has decreased. A total conception of 51% in 1955 has reduced to 38%. Does it speak we have to make use of modern technologies of ETT (Embryo transfer technology). The old conventional sleeve, rectal palpation, CL evaluation, reproductive disease diagnostic, has to be replaced with ultrasonography, laparoscopy, DNA guided diagnostic and improved fertility assessment technologies. Various modern technological impact on improving reproductive management have been discussed.
Uterine diagnostics for reducing (COD) Cystic ovarian disease, anoestrus, reproductive losses and infertility needs emphasis. The reproductive techniques consist of cryo-preservation, sperm sexing, spermatogenesis, invitrofertilization, nuclear transfers and introduction of disease resistance and animal welfare too. The reproductive difficulties in artificially produced calves, and stress to their mothers are averse to animal welfare. Various methods and their effects have been reviewed.
A new role of foeto maternal effects of hybridization of breeds has come to surface. Various such events have been expressed. New reproductive concepts such as Nitric oxide as a signaling molecule, Endothelial, nitric oxide synthase and its effects on reproductive ability have been explained. New knowledge models, like development of ovine concepts, foeto maternal interaction, extra cellular hormone action, cytokines, relay signals and their conversion to monomeric and dimeric forms have been postulated.
Hypothalmic astrocyle concept model with its biological control system has been detailed. Major technological innovations by the author in last 2 decades have been shown as pictorial monographs. A new concept of Dairy economic Vission 2050 for India has been forecasted.
A global policy prospect of Livestock industry for India has been attempted to glow Livestock marketing competition. A possible mega role for small farmers enterprise with mixed farming modules is presented. New generated technology transfer, research needs and gaps have been identified for young researchers.
Introduction:
1. Total Quality Management
(TQM) is the current buzzword in the production of quality food animal products for human consumption. The TQM concept implies that appropriate attention is given to all facets of dairy herds management in an effort to satisfy the consumer and their needs. Maintaining good records and using computer driven technology is recommended. Goals that are realistic and made possible by adopting a reproductive health programme are:
i. Average calving interval between 12 to 13 months.
ii. Breeding efficiency below 1.5 services per conception and less than 30 percent returns after 60 days.
iii. Postpartum interval to first standing heat between 30 to 40 days.
iv. Postpartum interval to first breeding no greater than 55 to 70 days.
v. Repeat breeders of 8 to 10 percent or less,
vi. Abnormal anestrus after 60 days postpartum reduced to 2 to 15 percent.
vii. Abortions of 1 to 2 percent or less.
viii. Retained placentas of 5 to 10 percent or less.
ix. Metritis of 5 to 10 percent or less
x. Culling percentage for reproductive failure below 18 percent.
2. Reproductive Efficiency
The famous reproductive scientist and clinician Zemjanis, compared the reproductive efficiency attained in U.S.A during 1955 with that of herds maintained in 1985. The first service conception had decreased from 53.4% to 38.7 in 1985 and the corresponding figures for total conception rate were reduced from 50.7% to 37.7%. The percentage of open cows at P.D. examination increased from 22 to 27. Repeat breeding was around 15% calving interval was 13.2 – 13.4 months. The prenatal deaths increased from 3.4 to 9%. Thus decline of reproductive efficiency in 1985 or 2007 over past 3-4 decades was observed. Is it that technologies of 1955 are not able to deliver efficient reproduction in 2007 ?. During the period a huge new set of technologies came into existence. Is that we need to incorporate new technology refinement in herd management especially reproductive ability ?. Should we go beyond traditional rectal palpation or sleeves and visits to new monitoring and evaluation through ultrasonic or other new technology?. A variety of application of new technologies are needed for heat detection. CL evaluation, reproductive disease diagnosis, semen evaluation, pregnancy diagnosis, infertility and other reproductive losses. This is where we need “Reproductive health Management through new techniques”. (Zem janis, 1987)
3. Uterine Diagnostics
The Uterine Condition can be disgnosed by Rectal palpation, vaginal speculum examination, uterine swab culture, uterine biopsy, ultrasonic imaging of uterus and ovaries, and progesterone testing. Abnormal postpartum cows are identified based on uterine horn size and texture in addition to vulvar discharge. Cows identified as abnormal are treated, with intrauterine infusion of antibiotics, antiseptics, chemical irritants, prostaglandin injection, or a combination of these agents. Most cows (>80%) should ovulate by 30 days postcalving. Cystic ovarian disease (COD) may occur in 5 to 10% of cows postpartum and delay first ovulation and first breeding. Anestrous cows will have small inactive ovaries with little change over sequential examinations. Follicles <15 mm in diameter may be found on examination, but fail to ovulate. Diagnosis of pregnancy at 35 days has enhanced value compared to 45 days or later. Pregnancy exam at 35 days reduces days between breeding and the percentage of cows over 48 days between breeding. (Ferguson, 1995)
4. Reproductive Biotechniques:
Based on the progress in scientific knowledge of endocrinology, reproductive physio- biology and embryology during the last fifty years, new bio-techniques have been developed and introduced into animal breeding and husbandry. Among them are oestrussynchronisation / ovulation induction, artificial insemination, multiple ovulation induction, embryo transfer (MOET), in vitro embryo production (IVP) and cloning by nuclear transfer. The aims of these reproductive technologies were initially to speed up the genetic improvements of farm animals by the increase of offspring of selected males and females, and the reduction of the generation intervals. The techniques of cloning by nuclear transfer applied for experimental purposes, have the prospect of more practical implementation in near future. The uniformity of herds for an early management or for the multiplication of transgenic animals after gene-targeting shall be its outcome. Within the farm animal biotechnologies, animal welfare should receive special attention. Transfer of embryos into dairy cows may lead to more dystocia. Intrauterine insemination via laparoscopy or laparotomy with minimal anaesthesia by breeder organizations operates stress in animals.
Both multiple ovulation induction and embryo transfer are generally accepted technically. However the transfer of embryos after multiple ovulation increase in embryonic death, larger calves with longer gestation times, and more dystocia (van wagtendo et al 2000). Some of the biotechnologies used in modern animal reproduction management can be summarized and named as :
5. Reproductive Techniques:
Reproductive techniques have been reviewed, wani, 2004 and are briefly reproduced as :
i. Ist Generation Reproductive Technologies:
a. Artificial insemination technology
b. Cryo-preservation Techniques.
c. Sperm sexing
d. Opportune time insemination
e. Spermatogenesis.
ii. 2nd Generation Reproductive Technologies
a. Multiple Ovulation Embryo Transfer (MOET).
b. Embryo Development Blocks.
c. Embryo Flushing & evaluation.
d. Embryo preservation and culture.
iii. 3rd Generation Reproductive Technologies
a. Invitro fertilization (IVF)
b. Clonning Techniques.
iv. Future Generation Reproductive Technologies
a. Stem cell technology
b. Embryonic cell lines.
c. Pro-nuclear microinjections
d. Nuclear Transfers
e. Viral Mediated Transgenesis
v. Reproduction Biotechnology application
a. New animal products.
b. Profitable production traits.
c. Disease resistance
d. Animal welfare and protection.
Successful somatic cloned animal production has been reported in various domesticated species, including cattle; however it is associated with a high rate of pregnancy failure. The low cloning yield could possibly arise from either an abnormal and / or poorly developed placenta. In comparison to control cows, fewer placentomes were found in somatic cell nuclearrecipient (NT) cows at day 60 of gestation, suggesting a retardation of fetal /placental growth in these animals. NT cows not only had fewer numbers of chorionic villi but also had poorly developed caruncles. Macroscopic examination revealed a typical development of the placentomes in terms of shape and size. Histological disruption of chorionic villi and caruncular septum was found in NT cows. Of particular interest was that the expression of genes, as well as proteins in the placentome, was disparate between NT and artificially inseminated cows, especially placental lactogen (PL) and pregnancy associated glycoprotein (PAG). In contrast, prolactin-related protein (PRP-I) signals were comparable across cows, including NT cows carrying immotile fetuses. The expression of extracellular matrix degrading molecule, heparanase (HPA) in NT cows was divergent from that of control cows. Microarray data suggest that gene expression was disorientated in early stages of implantation in NT cows, but this was eliminated with progression of gestation. These findings strongly support a delay in trophoblast development during early stages of placentation in NT cows, and suggest that placental specific proteins, including PLs, PAGs, and HPA are key indicators for the aberration of gestation and placental function in cows. (Kruip and Van Re, 2008)
There is a large body of evidence demonstrating that, in comparison with in vivo programme controls, the size and weight of IVP calves is higher (30% over 50 kg), the gestation period is longer, the % dystocia and the incidence of caesarean sections is much high. The % abortions and perinatal death are also higher (Behboodi et al, 1995; Kruip and Den Daas 1995; Wagtendon-de Leeuw et al. 1998; 2000). In general the calves are less active (Reinders et al. 1995). In addition the % of hydroallantois and congenital malformation including abnormal limbs and spinal cords, is increased in IVP calves and lambs. Taking together these problems are defined as the large offspring syndrome (LOS) (Young 1998). (Farin & Farin; 1995) and Sinclair et al. (1997) found a differential growth of organs (liver, heart , kidneys and adrenal gland) after IVP. Postnatally too some IVP calves anomalies were observed. (deRoos et al.2000)
MOET, including synchronization and induction of oestrus and AI, as well as IVP,NT are sometimes with serious consequences. We suggest (potential) risks of biotechnologies for farm animal welfare should be systematically assessed. The type of research should be multidisciplinary and should make use of appropriate scientifically valid experimental designs and protocols. Results obtained accordingly in developing and using the safest biotechnological methods and procedures. Technological progress which is ethically justified and beneficial for society in general the scientific and agricultural community i,e made use of for increased productivity.
6. New Concepts
Nitric oxide as a signaling molecule:
Many years scientists in reproductive research talked of endothelium-derived relaxing Factor (EDRF). This was thought to be a signaling molecule. The discovery that EDRF was in fact nitric oxide (NO). Over 60,000 papers have been published in last 10 years on NO. Nobel prize of 1998 was on this discovery. Nitric oxide is responsible for neurotransmission, immune defence, cell death, (apoptosis) and cell motility. It has a half life of few seconds. Enzyme producing it are (NOS) nitric oxide synthases (Phil, 2007).
Nitric Oxide being a small molecule, diffuses rapidly across cell membrane. Its diffusion distances can be several hundred microns and its biological effects are mediated through a range of targets as haem groups, cysteine residues, iron or zinc clusters.
Nitric oxide (NO) is now recognized as an important intracellular and inter cellular messenger. It relaxes uterine tissues, by inhibiting uterine contractility. Thus, by reducing uterine movement it helps in uterine quiescence. This is habitual of progesterones and antignostic to PGF2 alpha. NO has also a role in ovulation by interplaying ovarian steriodogenesis and prostaglandins.
Progesterone synthesis is reported to have increased in NO dose dependent rats (Yuan et al, 1999) Nitric oxide decreases oestradiol secretion in rats. It is established that NO plays a role in endothelium dependent vascular relaxation, phagocytic cell cytotoxicity, cenbtral nervous system neurotransmission and uterine relaxation (Moha et al, 2001). Granulosa cells and luteal cells in the rat ovary synthesize nitric oxide. Preovulatory follicles in their theca and stromal cell layers have enzyme synthesis. Endothelial nitric oxide synthase (NOSIII) and inducible nitric oxide synthase (NOS II) are among these enzymes.
Nitric Oxide is a fast acting signal which has the capacity to block entrance of blood components into follicular fluid. New concept of NOS system operating in ovulation and follicular rupture is now postulated. NOS acts via vascular dilation and ovulatory leukocyte distribution. The reorganization of follicular rupture and formation of corpora lutea requires nitric oxide. Thus, NO is having a role in ovarian physiology and steriodogenesis.
A neuropathway may be evinced in future. There is evidence of hypothalmic NOS – containing neurons, that regulate LH. Moreover, NOS containing (NERVE) fibers have also been found in rats. Does NO act as a local modulator of steriodogenesis? as many of its actions are mediated by Iron-containing enzymes, like guanylate cyclase and cyclo oxygene. The upregulation of progesterone and down regulation of oestradiol production in ovaries is now ascribed to NOS – system. NO may act by binding to the iron-sulphur moiety of the enzymes involved in the process of steriodiogenesis and effecting their production. (Yuan et al, 1999; Motta et al. 2001).
7. New Knowledge models
a. Development of Ovine conceptus
? Protective covering on caruncles
? Protective mucin covering.
? 18th day transitory attachment.
? 39th day cotyledons appear.
? 40th day projections from elongated Blastocysts into caruncular Epithelia.
? 42th day syncytial formation.
? 48th day attachment fixed as placentomes.
? Non-invasive attachment.
? Invasive attachment post 40th day.
? 80th Day of Gestation – placentomes, - structured appearance of recognizable individual cotyledons & caruncles disappear.
? Intra caruncular and caruncular cotyledonary attachment is supported by evidence from:
• Histological change
• Progesterone / Estrogen receptors
• Biochemical & Morphological evidences.
(Khatoon, 2007 – M.V.Sc thesis)
b. Foeto maternal interaction Transitory attachment
Binucleate cells (Gestation day) 18
Proliferation of Endometrial cells
E2/P4 receptors
Growth factors.
Progesterone receptors increase with gestation progress.
? Estrogen E2 receptors almost constant during pregnancy.
? Increased E/P ratio.
? Fluctuations in mineral, glucose, enzyme and proteins.
? Vaginal, cervical and uterine epithelial changes observed as pregnancy progressed.
C. Extracellular hormones actions (Concept by Canadian Scientists)
Growth factor or Cytokines …… Transcription of genes through specific receptor ….. Intracellular signaling molecules (Groner, 2002).
Conversion Monomeric to Dimeric form
Modus: Tyrosine phosphorylation
D. Hypothalmic astrocytes Employ ….. Transforming Growth Factor (TGF) Directly to regulate GnRH ….. controls … Sexual maturation … & …. Adult reproduction functions
This is what we name now as: Astrocyte – GnRH – Neuron signaling Pathway
Techniques used : Situ hybridization immunohistochemistry … … Motion communication processes.
Bouret et al, 2004 France
E. Biological Control
Multiple signals …. Receptivity or un-receptivity (Ringo et al, 1996 – U.S.A) Hypothalmo –pituitary – gonodotrophic – axis …. Mid gestation develops in sheep - late gestation … GnRH neuron centers develop in pregnancy or during foetal life … Only Maturational changes occur at Puberty – so as to form a GnRH neuronal system.
F. Hypothalmo – pictuitory – somatorophic axis begin in fetal life
Midgestation …. But Growth hormone R.H neuronal system does not develop until 10 weeks after birth. (Polkowska, 1995) Poland

8. “Major Technological Advances by author in last 10 years”
i. An attempt using ovaries and sperms from slaughtered sheep for invitro -fertilization and invitro-maturation techniques were used to produce morula. fig.1. These techniques innovated and published have aroused world wide recognitions, as awards. This work has been cited by Histcite-Index Lan wilmut founder of Dolly, the Sheep. (wani & wani,2000, 2003). In this era of biosafety and international Zoo and phyto sanitation, an attempt was made to limit bacterial and microbial loads in semen. Invitro bacterial sensitivity and bacterial load of semen was reported, (Seh et al 2000) (Koul 2002) which stimulated a new era of biosafety of gametes and higher fertility rates in cattle.
ii. The hitherto untapped research area of pre-natal development of ovine foetus was attempted (Mufti et al 2000, Wani and Buchoo, 1990). The investigation aroused much interest and citations. Very few studies on this area are available in world. This study is of great value for human and animal health. It has been further investigated during last few years. For the first time in world we have presented the mammalian foeto maternal union in its true form. This is an indirect association. A new concept of foetal growth, differentiation, foetal cotyledons within the elongated blastocysts has aroused the hope for preparation of a Invitro-Foetal –Development Model. (Wani,et al, 2006a, 2006b). These new concepts were presented in International conferences on Genomic and Proteomic concepts. The models were highly appreciated for use in various drug – gene – disease control interactions. (Wani et al,2006c; d;e). The binuclear and hybrid nuclear cells have been identified to specify syncytia & related mechanisms of foeto-maternal attachments. Fig.2. These concepts shall revolutionize disease resistance, control and prevention.
iii. We tried fish ovarian extract ( waste material) to replace costly synthetic hormones. This is ready for patatening. Laparoscopy for ovarian prediction along with ultrasonic use for pregnancy diagnosis in Sheep & goats was innovated and perfected . (Wani et al, 1998; Mufti et al, 1998).The use of hcG for improving lambing rates (Wani et al 2000) and use of laparoscopy for ovulation detection are new replications of our earlier investigations (Buchoo et al 2000).World famous Elsevier / Academic Press invited me to write a paper for Encyclopedia of Dairy Sciences Vol II PP 1259-1270. The only Asian to be invited to contribute on Goat Management Systems is in itself significant achievement summed up by the world famous Prof. John R.Campbell, the president Emeritus of Oklahama State University as “World renowned disciplinary expert, Wise and excellent author”. Our work on Economics of Pashmina goats and their Genetic characterization has been applauded by world famous Scientists and has helped Pashmina Industry in Ladakh to produce fine pashmina.
iv. Our recent prenatal and research on foeto maternal interactions is one of its unique in the world. Various gynaecological land marks, placental and immunological barriers, caruncle, cotyledon development and their association is of unique nature and can be reviewed in Fig 1-10.
9. Dairy Economics:
Dairy Farming in particular and Livestock enterprises in General suffer from two problems:
1. Higher production and maintenance cost
2. Low market prices of livestock products
Global milk pricing varies with Dairy Farm Structures. Dairy Farm Structures are either small 2-5 animal farms as ours or large more than 50 or 100. The countries like USA, EU, Canada, Isreal, Hungry, USA, South America and Oceania have large farm structures. Except Oceania none produce milk at world market price of 15-22 dollar a quintal. Thereby, signifying large farm structures are not always economical. Our small holding size of dairy structure is more competitive than West. This advantage needs to be harvested under WTO-Liberalization Global System. Should this need strengthening of small farmer’s co-operative? Do we needs implementation of small farmer friendly policies. The sale of 1 litre of milk in India is less than a litre of water. This needs serious consideration by our planning commission.
a. Higher Production Costs
The costs of milk production are high in Scandinavian Countries and Spain. The yesterday giants are todays loosers. The highest production price is also in counties like Hungary, Czech Republic, Isreal, Bangladesh, Thailand and parts of China. Their cost of production is more than global average cost of production viz 15-22 dollars/quintal. Those who produce at world market prices of 15-22 dollars include Estonia, Poland, Chile, Brazil, India, Vietnam and Oceania. Thus, we have tough competition with these nations to gain export markets. The Switzerland, Norway and Canada produce milk at much higher price than EU and USA, their markets in middle east are our export pastures.
b. Lower Milk Prices
The low milk prices than world market averages are in Argentina and Pakistan. However, they have distribution advantage rather than production axis, as farmers demorcating forums are non-existant. As such they may not pose any competition with us. We have export advantage in Asia as our other neighbour China too produces milk at higher costs. We must prepare ourselves as “Future milk Exporters in Asia”. This needs a preparation for zoo-sanitation and export oriented shift in policy decision.
c. North-West-Export-Zone
The northern-west states have self sufficiency in milk. We have to make this North-West-Zone comprising of J&K, H.P., Haryana, Punjab, Rajasthan and Gujarat as our potential milk export-zone. We will have to induce mission modes for its animal disease-free-status, so that we meet international-zoo-Sanitary standards. Poor zoo-sanitation is our major block to exports of milk. Outreach to farmers on these lines is suggested.
10. Global meat industry:
The next thrust area under transfer of technology has to make India “major meat exporting” country. The global meat industry is slowly moving in the hands of Brasallians and Chinese farmers. Thanks to our vegetarian majority diets we too are on run. Against, daily needs of 25g/person/day we produce only half of it on per capita basis, but our advantage with goat meat abundance can differentiate our export potential from the rest. Even global super powers USA have more demand for Cheron than what is produced. A policy change and frame work has to be drafted. Our product technology packaging and consumer preferences have to be webbed through transfer of technology to export concerns in particular and in general to home users. The focus has to be centered around hygienic meat production, ecological preservation, processing and product technology. This in principle means “reshaping livestock range and forage management” through effective communication and transfer of technology across its terminal and potential users and beneficiaries.
11. Generated Technology Transfer
We have to lead in Embryo transfer technology in Asia as has been done by us in dissemination of AI technology, in collaboration with Swedish expertise. Many international experiences are needed in this and other Transfer of Technology missions. We need bio-technological links to be developed with industry and user departments. Licensing in service mode is desired in technology transfer. We can do it on Cornell University pattern. Strengthening pre-clinical, clinical and basic diagnostic skills to glow animal science Institutes as a word referral clinical centers. This would need emphasis on technician training facilities for physiology, Reproduction, AI, Biochemistry neurosciences, biology and like. Linking, research and extension, faculty, students and farmers under KVK pattern are needed for quick animal science technology transfer. The referral TOT in animal sciences is ready to be switched on in the following area in next few years.
? Rapid diagnostic tools and techniques
? Tannin degradation technology and low- cost feed packaging
? High security disease prevalence checks. May be its sharing of emerging disease pattern with public health experts give us better dividends.
? Biotechniques for germplasm propagation and improvements.
12. In short we need Emphasis on
? Global-food- system-chain-factors(GFSCF)
? Research on competitive comparative health and disease (CCDH)
? Internationalize Veterinary and animal science education.
? Out reach for global farming through TOT.(Transfer of Technology)
13. Future Research Needs or Gaps
• Avoiding emerging diseases threats
• Refined disease diagnostics
• Drug – disease – targeting.
International Zoo-Sanitation regulations and product safety measures to fit us in Global – food – chain – operative systems.

14. Future Research Targets
i. Targeted gene delivery.
ii. Neuroregulatory mechanisms.
iii. Innate immune system.
iv. Protein interactions – transcriptional conformational mechanism.
v. Gene – drug – designing. Molecule targeting under Genome.
vi. Natural receptivity associated proteins for immune protection imaging and mapping.