1 Workshop on Economical Important Diseases of Buffaloes YASHDA 25 TH March 2009 Regional Disease Diagnostic Laboratory ( Western Zone ) Disease Investigation Section, Aundh, Pune – 411 007 MAHARASHTRA 2 INDEX Name of Author Sr.No. Name of Article 1 Economically important diseases of buffalo and their impact on export of buffalo meat Prof. P. K. Uppal 1 TO 9 2 Bubaline disease investigation: Current Status and Future Prospects P.D. Deshpande 10 TO 17 Raut CG 18 TO 23 Markandeya , N. M 23 TO 43 3 4 Present status of buffalopox in India Non-specific infectious infertility in dairy animals: Impact and control measures Page No 3 Key Note Address. Economically important diseases of buffalo and their impact on export of buffalo meat. by Prof. P. K. Uppal B.V.Sc & A.H., M.V.Sc., Ph.D (Agra), Ph.D. (Cantab) U.K., FNAVsc, FIAAR Technical Director Diagnostic Research Laboratories, RWITC, Ltd. (Approved by Govt. of India) 6, Arjun Marg, Pune – 411 001 India has immense potential for export of buffalo meat owing to its large buffalo population. Indian buffalo constitutes about half of the world buffalo population and accounts one third of the total cattle population in this country. It contributes about 50% of total milk production of India and provides more than 20% of the meat production. The meat export suffers sometime because of diseases because importing countries raise certain objections for diseases like foot and mouth disease, sarcocystosis, brucellosis, tuberculosis and food pathogens like Campylobacter jejuni, salmonella typhi, shigella dysentariae, staphylococcus aureus, E.coli, Listeria monocytogenes and Yersinia enterocolitica. Buffaloes in India are raised by millions of farmers in unit of 1 – 3, who are primarily small and marginal farmers. They mainly exist in Punjab, Western Uttar Pradesh, Haryana and Delhi. The export of meat was initiated in early 1973-74. During that period, 1920 tonnes of buffalo meat was exported out of 2005 tonnes of total meat exported. Former Managing Director, BIBCOL. Dept. of Bio Technology Phone: Pune: 0091-20-26362666/ 26360884 Fax: 0091-20-26333129/26361036 4 Email: profpkuppal.vsnl.net Export was targeted to Persian Gulf and Middle East countries and showed rapid growth and the export reaching a level of 54582 tonnes during 1981-82. The meat export suffered a setback during 80s as a result of ban imposed by Saudi Arabia and other Gulf countries on account of Rinderpest and foot-and-mouth disease. The bulk of buffalo meat is exported as deboned, deglanded frozen meat which accounts for 75% of the total export. The major markets presently are Malaysia, Philippines, UAE, Iran, Oman, Egypt, Kuwait, Mauritius, Jordan, Bahrain, Yaman Arab Republic, Greece and Turkey. Global arrangements covering the meat trade have been set by World Trade Organisation (WTO). The requirements for exporting meat include standards imposed for quality, price, and punctuality of delivery. For Middle East countries, the basic regulatory requirements are Halal slaughter and the fitness for human consumption certificate. Zoo sanitary requirements have become more and more stringent. It is the primary responsibility of exporting countries to take appropriate steps to minimize the possibility of disease transmission through meat export. The following issues, which form the basic constraints were identified. 1. The management of economically important diseases listed in OIE Manual from Zoo Sanitary standards of meat exports. 2. Creation of a Federal Zoo Sanitary Service, Certificate control and Quarantine service. 5 There is a need for effective utilization of buffalo resources for sustainable buffalo production of meat, milk and work. There is a tremendous loss of buffalo germplasm through city dairy through disposal of productive animals for slaughter. In all, more than 7 million male calves are produced every year and less than half are put into agricultural transport mainly in sugarcane belt. This precious germplasm could form a basis for strong meat industry alongwith raised buffalo broilers if proper infrastructure is created for fattening them. Above all, it would be pertinent to develop animal healthcare plan for complete freedom of certain diseases specially foot and mouth disease, and all important diseases indicated by OIE. Therefore international standard meat processing plants for processing is a must. In addition buffaloes are to be raised in disease free zones so that the export could be managed profitably. Above all animal identification and traceability of buffaloes for export of meat will be one of the requirements for better competitiveness. ANIMAL HEALTHCARE PLAN: In India, rinderpest was a main threat for the export of buffalo meat but tremendous efforts made by animal health scientists paid dividend and the disease has been declared free by OIE in India. This paved way for the export of buffalo meat to various countries. However, there is a need for other healthcare parameters including various contagious and infectious diseases of buffaloes. For better productivity the diseases of concern in buffaloes are Haemorrhagic Septicaemia (Pasteurellosis), Black-quarter Foot and Mouth disease and Brucellosis. Similarly other diseases which have a 6 role on the productivity of buffaloes are Tuberculosis , Johne’s Disease, Listeriosis, Tetanus, Salomonellsis, Blue tongue, Bufallo fox and Infectious Bovine Rhinotracheitis. Animal health is the foundation to animal production. Therefore, country demands multi-pronged attack for buffalo production programme in order to reach the goal of healthy production cycle of buffalo – the species which is bestowed with immense potentialities for meat and dairying industries. There are certain pathogens which influence the quality of meat and their presence in meat may cause diseases in man and therefore may become liable for rejection for human consumption. The discussion would be concentrated on problems associated on buffalo health and food borne infection and intoxication associated with buffalo meat will be discussed. FOOT AND MOUTH DISEASE: It is an extremely contagious, acute disease in buffaloes which is characterized by fever and vesicular eruption in the mouth and in the feet. In India, there are three strains , Asia I, O and A. There are different strains with different degree of virulence. The virus seems to be capable of infinite mutation so that new antigenically different strains are constantly appearing. The difficulties therefore are encountered in vaccination programme. The disease is well documented amongst buffaloes in India. The disease results into loss in milk production and productive capacity of buffalo. Export of buffalo Meat suffers many a time because of foot and mouth disease in the country. Therefore control of foot and mouth disease in India is given top priority. In order to boost meat export OIE agreed to have FMD free county or even zone with or without vaccination. 7 Countries like Argentina , Botswana, Taipei China, Malaysia and Uruguay. The OIE requirement for the export of meat from FMD free zone where vaccination is practiced requires following practices. 1. Prompt animal disease reporting. 2. No outbreak of FMD for the past 2 years and no evidence of FMD virus infection for the last 12 months. 3. Regulatory measures for the prevention and controlled of both FMD and FMD virus infection. 4. Supply documented evidence that appropriate vaccine is used and there is frequent surveillance. 5. There is need of international veterinary certificate that animals had been slaughtered in approved abattoir and have been subjected to anti-mortem and postmortem inspections. In order to control foot and mouth disease in India purified vaccine application will be useful. Moreover NSP test will be helpful in vaccinated population under field conditions. BRUCELLOSIS: Brucellosis has been recognized as a disease of major economic importance in buffaloes and it may occur through buffalo meat. Brucellosis is caused by a Gram negative bacteria named as Brucella abortus. Brucella is a highly resistant organism and it may be transmitted through contact of meat through fomites/flies or waste disposal or by consumption of insufficiently / partially / uncooked meat. 8 It is mainly caused among the workers at slaughter house or those processing the meat. In India, where slaughter of buffalo is done in unorganized sector, there are of every chance of Brucellosis to be transmitted to the handlers who are slaughtering / processing the meat. In India, lot of nonproductive buffaloes are responsible for transmission of brucellosis. It is pertinent that for eradication of brucellosis, quick diagnoses with more specific and sensitive tests are adopted. It is advisable that buffaloes must be tested for brucellosis before slaughter and would help better export and will prevent the transmission to human beings. The best approach to control is to have effective vaccination adopted by test and slaughter programme. INFECTIOUS BOVINE RHINOTRACHEITIS Bovine herpes virus 1 is an important viral pathogen cause respiratory infection, pneumonia, conjunctivitis, severe abortions, vulvovaginitis, balanoposititis and Immunisuppression. The disease has a high morbidity and low mortality. The virus of this causes latent infection which can persist for life and is a major problem of buffaloes bull. The disease is wild spread. The is difficult to diagnosis on the basis of clinical symptoms. The method used for laboratory confirmation include virus isolation fluorescent antibody test, ELISA, virus neutralization test and histopathology. The appropriate vaccine in the country is the need of the hour which is in the process of development. Hygienic semen collection free from IBR is important for better productivity of buffaloes. 9 SARCOCYSTIS: In India, incidence of sarcocystosis is found to be quite high in cattle and buffaloes. The highest percentage of infection was observed in oesophagal muscle, followed by shoulder and diaphragmatic muscle and lowest in heart muscle. Sarcocyst is a coccidian parasite with carnivores as definite host and herbivores (cattle, buffalo) as intermediate hosts. The meat containing sarcocysts is condemned from aesthetic point of view. The species involved are Sarcocystis bovicanis and Sarcocystis hominis. The trichinoscopic diagnostic methods, techniques histological are visual method examination, and presently, immunodiagnostic techniques like indirect haemagglutination microtitre test, indirect FAT and ELISA. FOOD SAFETY Because of consumer awareness, there is a need for national food hygiene development programme, which should include microbiological hazards analysis in buffalo meat. National assessment of pathogen- commodity combination will achieve one of the priority area of the work which is advised by the Codex Alimentarius Commission (CAC). Due to Threat Perception on the spread of animal and zoonotic diseases through buffalo meat it is essential to observe certain disease control sanitary measures like Hazard analysis Critical Control Point (HACCP) Good Manufacturing Practices, (GMP) and ISO standards. 10 Following issues need to be addressed for the safety of Buffalo meat and meat products. 1. Food Production Chain has to maintain quality and safety assurance system in the entire livestock product chain from Farmer-to-consumer, to ensure that livestock products have systems based control which meet national and international standards. Some of the measures are hazard analysis Good Manufacturing Practices (GMP), Good Agricultural Practices (GAP), critical control points (HACCP), and cold chain. 2. The control process should be consistent with the standard set by the Codex Alimentarius and the Government should develop the guidelines. 3. A change in focus from individual animals to herds and population for better economic return for meat and meat products to monitor better health status. 4. Development and implementation of new technologies for food and feed production, preservation and commercialization, and related problems of toxic residues and improved standards of hygiene. 5. For market development, standards towards production of safe food to avoid rejection of Indian products due to use of immunobiologicals, antibiotics and other pharmaceutical. 6. Legislation is set up where ever necessary to maintain the standards. 7. Enforcement of legislation for accountability. 8. Setting of Structured laboratories for better monitoring plan for surveillance of food born diseases. 9. State veterinary services are empowered the official responsibility for surveillance and monitoring programme. 10. Education, training and public awareness in food hygiene. 11 11. Effective linkages with OIE, WHO, FAO and WTO to have trade intelligence ANIMAL IDENTIFICATION AND TRACEABILITY Animal identification and traceability are closely related to disease control procedure. They are epidemiological tools that have a major impact on matters such as public health and trade. However country like India can benefit immensely to export buffalo meat if they adopt identification system in a zone or a region which is free from foot and mouth disease. This concept is fundamental animal disease control tool in the context of regionalization and compartmentalization. ACKNOWLEDGMENT Thanks are due to Disease Investigation Section for inviting me to deliver the key note address in the workshop to be held on 25th March 2009 at Yashada, Banner Road, Pune. 12 Bubaline disease investigation: Current Status and Future Prospects P.D. Deshpande, Nagappa Karabasanavar, R.P. Kolhe and A. R. Ratnaparakhi Department of Veterinary Public Health KNP College of Veterinary Science (MAFSU, Nagpur), Shirval, District- Satara, Maharashtra- 412 801 Modern day domesticated water buffalo (Bubalus bubalis) is a cloven footed animal belonging to the bovine group. Other related members of this group include lowland anoa (Bubalus depressicornis), tamaraw (Bubalus mindorensis), and mountain anoa (Bubalus quarlesi), etc. Basically, members of buffalo fall under two categories i.e. African wild buffalo (Syncerus) and Asian buffalo (Bubalus) of which the later eveolved as the domsticated buffalo today, known as water buffalo. Another similar species is bison (American buffalo). Buffalo are one of the most economic domestic animal species predominanlty found in Asia, Africa and mediterranean countries. There is an increasing trend in the world buffalo population, of the total 174 million heads india contributes 98 million (56.4%). Buffalo population wise india is followed by Pakistan with 26 million heads (15.1%), China 23 million (13.1%) followed by other south-east Asia and African countries. Buffaloes are basically reared in rural sector that provides basic livelihood in the from of milk and milk products (mozerella cheese), meat (beef or carabeef), draft (especially paddy fields), manure, fuel, hide, etc. However, there is increasing trend in the urban buffalo population (due 13 to high total solids in buffalo milk, more number of value added / byproducts can be manufactured and buffalo milk fetch more market price due to fat). Today, buffalo has emerged as an indispensible to the Indian milk industry by producing 5740 thousand tonnes of milk (year 2004); that contributed 66.3% share of the global buffalo milk productuion (76548 thosand tonnes). As par as the global meat inventary is concerned the buffalo constitutes a meagre of 1% share, while in India both cattle and buffalo put together constitute a considerable 32% share of national meat production. Buffalo: Still wild in instinct! During the process of evolution that proceeded with domestication, the wilder instinct of the buffalo (prey-predator behavior) continued to persist. Hence, buffaloes exhibit no overt clinical signs though a progressive disease is existent in the body, which poses difficultly in the diagnosis. In order to understand the physiopathology of a disease / disorder in this particular species necessitates a rational thinking. Hence, disease diagnosis aims at early and accurate diagnosis so that production, performance, and reproduction are restored or enhanced. Modern day diseases are complex and multifactorial in nature. Diseases with multiple etiology are precipitated by endemicity of the pathogens, production stress, immunosupression due to pollution & parasites, iatrogenic problems, etc. For which the managemnetal strategies include exact diagnosis, selection of effective drug, complete treatment regimen with supportive therapy for successful treatment in order to restore health. The health and productivity of an animal basically depends upon its genotype and the environment has a profound influence on it. Hence, for the better productivity, selection of elite and tested stock is essential. In the modern, intensive type of buffalo farming the husbandry aspects 14 are to be given due credit. Especially with respect to animal density which in an ever increasing threat to its health and productivity. The ever increasing human population has also put demanding restriction especially migration on animal species including the buffalo. Through migration the new incumbents bring newer disease to the naïve area. Similarly, the product trade which translocates not only within the states but also across the international tranasboundaries. Both of these domestic and international animal or product mobility has consequences which demand a robust diagnostic approach and a system to address. Prompt diagnosis: Prompt diagnosis of a disease rests on clinical symptoms, laboratory findings and expertise. However, in buffalo the overt clinical manifestation are often mild or non-existent. For such sub-clinical representations a robust laboratory investigation is a must. In this direction laboratory examination of blood, milk, and other excretion / secretions of an animal are of paramount importance. Further, in order to aid the routine therapeutics testing of the organisms for the drug of choice is also equally important. Hence, drug sensitivity testing should become a routine for the practicing veterinary clinicians. Especially, the production related problems such as mastitis, anestrous, infertility should never be ignored. Major bubaline ailments: Following is a list of major ailments that buffaloes often suffer from. Which require laboratory aid for the diagnosis and management viz. arthritis, autoimmune diseases, bloat, brucellosis, diphtheria, calf scour, dermatitis, diarrhoea Johne’s disease, BVD, calf , FMD, IBR, infertility, joint ill. ketosis, lameness / laminitis, listeriosis , lumpy jaw, lungworm, mastitis, metritis, navel ill, neoplasms, poisoning, pneumonia, pox, prolapse, ringworm, rotavirus, traumatic reticulitis, T.B, wooden tongue, etc. Of these here is a list very important infections 15 that need critical attention include anaplasma, anthrax, babesia, BQ, brucellosis, coccidiosis, ehrlichiosis, fascioliasis, HS, FMD, rabies, surra and theileriosis. Diagnostic services: In order to cater the basic needs of the public, there is a need to establish disease diagnostic centers. Preferably, a well equipped laboratory within a radius of 20 km animal disease diagnostic lab and a regional lab for investigation of epidemics. Such centers should have ties with nearby institution such as veterinary colleges or national institutes and referral laboratory. These diagnostic facilities generate information on health and production. Thereby a national repository or database could be created which help even in the early warning systems of diseases. Also, these diagnostic faculties management, help in streamlining formulating of national strategies and local for disease programs (documentation, traceability and responsibility) and help the policy makers to take appropriate decisions. Further, the establishments help to control and regulate the local, domestic as well as international trade in livestock and its products. Rationale: The disease investigation is an integrated and systemic program. It involves following aspects • Diagnosis • Field investigation • Disease management program • Training and research • – Students – Technical staff – Field practitioners Improvement of diagnostic techniques and procedures 16 For the accurate diagnosis of a disease the sampling method and techniques are very essential. Various types of sampling are employed for the field investigation, for instance probability or non-probability; individual or mass, etc. However, there are certain aspects that should not be overlooked i.e. prevalence, population size, precision and confidence level. Diagnostic tools: The diagnostic approaches are basically directed towards the remediation of production, disease and reproduction. After ascertaining the underlying cause through prompt diagnosis the therapeutic aspects become easy in restoring the productivity. For the diagnosis of bubaline ailments efforts are directed towards the diagnosis of clinical, sub clinical and latent infectious agents, noninfectious / metabolic disease or disorders and also in zoonoses surveillance. There are various areas of diagnostics depending upon the type of investigation required. • Pathology : clinical, RBC/ WBC count, differential count, hematocrit/ hemoglobin Count, histopathology, staining, etc • Microbiology / bacteriology : bacterial isolation/ identification, antibiotic sensitivity test, food/ water analysis, fungal detection, etc • Immunodiagnosis / serology : Rapid Plate Test -RPT, Tube Agglutination Test, Agar Gel Precipitation Test-AGPT, EnzymeLinked Immunosorbent Assay –ELISA, Complement fixation testCFT, Hemagglutination-Inhibition Test -HA-HI) • Virology (isolation/ identification, egg / lab animal inoculation, tissue culture, serology, etc • Parasitology : routine blood examination i.e. direct smear, special staining; and fecal examination i.e. direct smear, flotation, sedimentation, larval culture, etc 17 • Toxicology : mycotoxin analysis, other toxicological examination, toxicity Test , heavy metals, pesticides, veterinary drug residues, growth promoters, food additives, preservatives, etc • Molecular biology : techniques such as PCR (RT-PCR), RFLP, etc offer great advantage over the other techniques by virtue of rapidity, sensitivity, specificity, economy, repeatability, reproducibility, differentiation, etc The use of such diagnostics rest in the diseases management strategies so as to improve the buffalo productivity and sustainable animal husbandry • Early and accurate diagnosis of a disease leading to successful and economic treatment of animal and restoration of productivity. • Isolation, segregation and quarantine of cases which helps in limiting a disease. • Gives an idea of specific environmental sanitation approves thereby pathogen load is minimized and nullified . • Mass treatment and vaccination strategies can be defined. • Biosecurity, surveillance and legislation processes are hastened. • Research, development and training are strengthened. • Local, national and internationally notifiable diseases are reported. • Such approaches help in the creation of disease free zones leading to disease eradication. Future prospects: Although consequent to the improved quality of human life with the advent modernization and mechanization, still there is a threat of zoonoses due to population dynamics, climate change, pollution, animal / product trade, globalization, etc. hence, zoonoses surveillance should be an integral part of a diagnostic facility. Productive and reproductive health of buffaloes should be routinely monitored, as they show silent heat and often do not show clear-cut 18 clinical manifestation. Hence, for such sub clinical ailments they should be routinely screened. Also, surveillance for repeat breeding, infertility, mastitis etc need to be undertaken. In the post- Mumbai terrorist attack era, the chances of bioterrorism can not be completely ignored. Hence, we working in the area of animal science should be prepared to tackle those infection / intoxications even not existent in our land. For this purpose, disease diagnostics have a pivotal role to play. Laboratory requirements: Keeping in view the hazards to the investigators, operators, assistants and other supporting staff in the laboratory, it is essential to take utmost care to ensure the occupational safety. Also, disaster preparedness should be scrupulously observed. The laboratories involved in diagnosis should possess required specifications. They should also be accredited. Relevant certification (ISO, GLP, etc) are to be followed and the laboratories should be in compliance with the national norms. All these provide adequate safety thereby enhancing the throughput emerging from its premises. Conclusion: Buffalo continues to be one of the major animal species of India involved in the rural upliftment, poverty alleviation, livelihood earning and foreign exchange. However, there are certain deviations (disease, disorder) that affect its optimal performance, production, and reproduction. Being still continuing with its wild instinct buffalo often does not exhibit such deviations (if not always, occasionally). Hence, there is a need to correct such deviations through skillful management that are often not detected by even the most experienced veterinarian. The laboratory diagnosis is therefore of paramount importance. In India, although there are many such laboratories (state and central Govt., as well as private), but they are not at present meeting the demand. So, 19 apart from government installations such as SVUs/ SAUs, ICAR (IVRICADRAD, PD_ADMAS, etc), DRDO, etc there is a need to encourage more private diagnostic / public testing laboratories. Such laboratories should have sufficient infrastructure, specifications, skilled man-power and government support. They should be accredited, certified and should comply with specified norms (GLP, GMP, ISO, etc). Productivity of an animal lies in its positive health; hence these diagnostic labs shall cater such field requirements. Better animal health ushers better quality living, buffalo production in particular would definitely be benefited by it, thereby paving ways for a developed nation ahead. 20 Present status of buffalopox in India Raut CG; Suryavanshi SN*; Raut WC**. National Institute of Virology, 20-A, Dr. Ambedkar Road, P.B.No.-11, Pune-1; *Disease Investigation Section, department of Animal Husbandry, Government of Maharshtra, Aundh, Pune-7. **Padmashree Dr. D.Y.Patil Medical College, Hospital & Research Centre, Sant Tukaram Nagar, Pimpri, Pune-18. Disease: Buffalopox: It is a viral disease characterized by typical pox lesions mainly on udder, teats, buccal cavity, ear, eyelids and to some extent on other parts of the body. Economic Impact: If the lesions on udder and teats in severe form, milk yield goes down up to 50-80%. Because of the secondary infection animal may develop mastitis and sometimes become unproductive. Due to zoonotic nature of the disease, there is a loss in manpower hours for certain period. Etiology: Agent: virus, Family: Poxviridae; Genus: Orthopoxvirus – Members: as follows Poxvirus Taxonomy: Orthopoxvirus: 1. Camel pox virus, 2.Cowpox virus, 3. Ectromelia virus, 4. Monkey pox virus , 5. Raccoon pox virus, 6.Tatera pox virus, 7.Vacciniavirus. i)Vaccinia virus Ankara, ii) Vaccinia virus Copenhagen , iii) Vaccinia virus Tian Tan, iv)Vaccinia virus WR, v) Buffalo pox virus, vi)Rabbit pox virus Utrecht, vii)Cantagalovirus 21 8. Variolavirus, 9.Volepoxvirus Genome:DNA , size : 300x100 nm , shape: brick shaped. Geographic Occurrence: Brazil, Egypt, Iran, Pakistan, Bangladesh, India. In India: Haryana , UttarPradesh, MadyaPradesh, Maharashtra, Gujrat, TamilNadu, In Maharashtra: Dhule, Jalgaon , Beed, Latur, Nanded, Usmanabad, Aurangabad, Nashik, Solapur, Kolhapur. It occurs generally during late winter and early summer. In India, buffalopox disease reported since 1937 till date with less to severe morbidity. There were number of viral isolations from vesicular fluids and skin scabs from humans and buffaloes. Disease in Animals: Lesions on teats, udder, ear, eyelids with some generalized form. Some cases were reported pox lesions with otitis and conjunctivitis with purulent discharge. No mortality in adults but in calves may be due to anorexia as the lesion may be on the buccal mucosa. 22 Disease in Humans: Fever for 2-3 days, pox lesions on hands, pain at site and corresponding axillary lymphadenitis. Generalized one or two lesions may be on the face and leg. Adults as well as children may get affected. Transmission: Close contact with infected animal. Animal handlers generally affected by close contact with the lesions. Mechanical transmission may possible by flies and insects. Animal Models: Rabbits, Rats, Mice Experiments conducted in chick embryos, albino rabbits and infant mice; and electron microscopic observations revealed a pattern of morphogenesis similar to that of vaccinia virus, except that the extracellular forms of the virus were devoid of well defined lateral bodies. Study conducted by intranasal instillation of buffalopox virus in infant 23 suckling mice produced fatal disease, which can be used as an animal model for studying pathogenesis of buffalopox virus. Diagnosis: On the basis of clinical picture: typical pox lesions on the body Genome detection by Polymerase Chain Reaction (PCR) & analysis by sequencing Virus particles detection by Electron Microscopy by negative staining of vesicular fluid, or microsectioning of scab. Virus isolation: In ovo: (embryonated egg inoculation)- choreoallontoic membrane (CAM) In vitro: (cell culture) BHK-21, Vero. : look for cytopathic effect In vivo: scarification of shaved skin of rabbits: look for development of pox lesions Treatment: Generally symptomatic treatment is advisable. Cleaning & washing of infected as well as normal animal during outbreak with 1% potassium permangnate solution. Topical application of antibiotics on lesions. Broad-spectrum antibiotics to check the secondary infections. Control: Segregation of the affected animals and treatment to check secondary infections (particularly the mastitis) were recommended to curtail the disease. Restricted movement of affected animals. Animal handlers should use the gloves for washing , cleaning and during treatment of the lesions. Till now no vaccines are available. 24 Further readings: Wariyar KC. (1937)Variola in buffaloes. Indian vet. J., 14, 169-170. Maqsood M. (1958) Generalised buffalopox. Vet. Rec., 70, 321-322. Singh IP; Singh SB. (1967) Isolation and characterization of the etiological agent of buffalopox. J. Res. Ludhiana, 4, 440-448. Baxby D; Hill BJ. (1971) Characteristics of a new poxvirus isolated from Indian buffaloes. Arch. ges. Virusforsch., 35 (1), 70-79. Lal SM.; Singh IP. (1977) Buffalopox: a review. Trop. anim. Hlth Prod., 9 (2), 107-112. Chandra R., Garg SK., Rana UVS ; Rao VDP. (1986). Pox infection of buffaloes. Farm Animals, 2, 57-69. Chandra R., Singh IP., Garg SK; Varshney KC.(1986) Experimental pathogenesis of buffalopox virus in rabbits: clinico-pathological studies. Acta virol., 30 (5), 390-396. Mathew T; Mathew Z. (1986) Isolation, cultivation and haemadsorption of buffalo poxvirus on BHK-21 cell line from Dhule Epidemic (Western India). Int. J. Zoonoses, 13 (1), 45-48. Nedunchelliyan S, Reddy DS., Venkataraman KS. (1992) Buffalo pox infection in man. Ind. J. public Hlth., 36 (2), 57. Dumbell K; Richardson M. (1993) Virological investigations of specimens from buffaloes affected by buffalopox in Maharashtra State, India between 1985 and 1987. Arch. Virol., 128 (3-4), 257-267 Kolhapure RM., Deolankar RP., Tupe CD., Raut CG., Basu A., Dama BM., Pawar SD., Joshi MV., Padbidri VS., Goverdhan MK. & Banerjee K. (1997) Investigation of buffalopox outbreaks in Maharashtra State during 1992- 1996. Ind. J. med. Res., 106 (11), 441-446. Raut C.G., Tatwarti S.B., Deolankar R.P., Kolhapure R.M., Tupe C.D. (1997): Clinical report: Buffalopox zoonosis in Nashik suburb (Maharashtra State) Livestock International. 7 P18-19. 25 Ramanan C; Ghorpade A; Kalra S; Mann S. (2007) Buffalopox. International J. Dermatology 35(2)128-130. Raut CG; Kulkarni AM; Pathak SV; Potekar AM; Ramteke VG; Chikbire SB (2009) Outbreak of Buffalopox associated Zoonosis in Kolhapur district of Maharashtra. Compendium, Ninth Indian Veterinary Congress, XVI Annual Conference of Indian Association for Advancement of Veterinary Research (IAAVR) & National Symposium on Transition from Empiricalism to Molecularism in Veterinary Research for a sustainable Animal Health & Production- Way Forward, Bombay Veterinary College, Mumbai. February 20-21, 2009. P36. 26 Non-specific infectious infertility in dairy animals: Impact and control measures Markandeya, N. M. and Deshmukh, V. V. College of Veterinary and Animal Sciences Maharashtra animal and fishery Sciences University Parbhani - 431 402 (Maharashtra state) Sustainable modern scientific dairying is based on regularity and normalcy of animal reproduction. Fertility management in dairy animals is an important global issue. Interference in routine reproductive process by the diseases, poor nutrition, inadequate herd management, hereditary and congenital factors, hormonal disturbances or environmental changes, makes the animal temporarily infertile (Osmanu, 1979). Infertility denotes reduced degree of fertility and records failure of a cow to produce a viable calf in each calendar year regularly. Bovine infertility is potent hazard for genetic improvement and milk production in dairy animals. Cattle are deemed infertile when they are neither normally fertile nor completely sterile. Interest in bovine infertility increased with the introduction of artificial insemination in the 1950s and as the farmers, herdsmen physiologists and other workers experienced reproductive failures in high milk producing dairy animals under field conditions (Roberts, 1956). Sub-optimum fertility is the major problem in reproduction of dairy animals and hence, traditional dairy farming and sub-fertility are responsible to each other. Infertility problems can be classified by various ways but infectious and noninfectious infertility is the most important and practically essential criteria of infertility classification. Infections very commonly affect all stages of reproductive processes and are costly and time consuming. Specific and non specific infections are invading reproductive 27 tract very commonly and reproductive process is either delayed, stopped or altered by the organisms. Nonspecific infections It is necessary to understand difference between specific and nonspecific infection of reproductive tract. Nonspecific infection is the infection, which is not known to be caused by a specific pathogen. Non specific infection can be described as the condition relating to immunity that occurs naturally as a result of animal’s genetic makeup or physiology and does not arise from a previous infection or vaccination. The non specific pathogens are generally termed as wound invaders and many non specific organisms act as opportunistic pathogens. Non specific organisms are germs which are available any where in the universe including the bodies of live animals. Non specific infectious agents are influenced by perpetuating or predisposing causes in animals and they tend to affect only individuals, whereas, the specific infections are mainly of enzootic type. Non specific infection enters animal system by lateral or horizontal transmissions. Existence of certain micro-organisms is well documents in healthy genital tracts of dairy animals. Most of the infection in non gravid horns is transient and self limiting. Non specific infection leads to contamination of genital tract, followed by elimination and repeated contamination. Non specific infections reduce reproductive efficacy of dairy animals and also the profit potential of dairy farms. Contaminants of reproductive tract Non-specific infectious infertility is caused by bacteria in particular and other pathogens like viruses, fungi, protozoa and mycoplasma infections in general. Mixed infections are common in reproductive tract leading to non specific infectious infertility. Bacterial Pseudomonas organisms pyocyanea, include Pseudomonas Actinomycetes pyogenes, auregenosa, Klebsiella 28 pneumoniae, Corneybacterium pyogenes, Corneybacterium Suis, Corneybacterium diptheriae, Salmonella paratyph, A and B, Salmonella typhi, Neisseria gonorrhoea, Neisseria meningitides,, Bacillus anthiracis, Bacillus subtitis, Fusobacterium, Bacteriods,, Staphylococcus aureus, Staphylococcus albus, Streptococcus pyogenes Streptococcus faecalis, Streptococcus agalactiae, Escherechia coli, Proteus vulgaris, Shigella shigae and many more. The record of various bacterial organisms in reproductive tract is well documented in the literature. Auther et al. (1960} has recorded evidence of synergism between A. pyogenes and fusobacterium nechrophorum, the later organisms apparently produce leucocidal endotoxin which would interfere with the host’s ability to eliminate A pyogenes. Bacteriods spp. also produces substances that interfere with phyagocytic ingestion and bacterial kill in the genital tract. Bane (1980) reported the composition of uterine flora of healthy and abnormal puerperium and recorded that the composition was very similar but significant quantitative and strain differences exists in pathogenic conditions. Hussain et al. (1990) reported that no bacterial species was consistently reported from cows for more than 2 weeks of parturition. Williams et. al (2007) reported that Escherichia coli is the pathogenic bacterium most frequently isolated from the post partum uterus and is associated with increased concentrations of peripheral plasma acute phase proteins and fetid vaginal mucus. Mycoplasma is self replicating prokaryotes under class mollicutes and these organisms lack in cell wall and hence they are resistant to penicillin or its analogues. Mycoplasma bovis and ureaplasm have been reported in reproductive infections. Incidence of abortions due to mycotoxic infection in dairy herds is recorded by Rameshkumar et al. (1988). Mycoplasma is associated with infertility but their exact 29 aetiological role is difficult to ascertain because they are present in the tracts of healthy animals. Oestrogenic mycotoxicosis is a disease caused by fungal toxin which mimics the action of oestrogen and stimulates oestrus like activities like hyperemia and odema of vulva, mammary and teat enlargement, prolapse of vagina. Heat and humidity in tropical countries create conditions favorable for fungal growth. Fungi of Candida tropicalis, Candidia albicans, Aspergillus fumigatus, Aspergillus niger, Penicillium types have been reported to cause genital infections and related infertility in animals. Kodagali (1979) recorded candida, aspergillus, fuzerium, mucor species of fungi and yeasts on prepucial sac of breeding buffalo bulls and reported that loose prepuce or prepucial prolapse leads to injury and infection of prepuce in breedable males. Pathogenenic process of infertility Bacteria or pathogens are symbionts of reproductive tract that some become pathogenic when the animal is stressed where as others are immediately pathogenic. Inflammation of reproductive that in dairy animals occurs when physical and functional barriers are breached and organisms enters in the tract. Uterus acts as incubator as it provides regularly moist, dark, warm, sterile and facilitative environment with nutrients which favours harboring of microorganism for a quiet long period. Fortunately most of the animals do not develop severe genital infections. However, fluctuation in bacterial colonization in genital tract occurs regularly. Bacterial flora is continuously changing because of elimination and recontamination. The chances of non specific infections are more in rainy and winter season and also in cases of indoor calving. Non specific infections leading to infertility enter through ascending routes and 30 particularly due to wounds, lacerations of external genitalia as uterus is most vulnerable for the environmental infections especially immediately after calving or when afterbirths are retained for too long periods. Bacterial load usually reduces during first three weeks after partition but can last up to 14 weeks in some cows. Bacteria may interfere fertility in animals by directly killing gametes or concepts, through change in uterine milk composition, by causing inflammation or by cause chronic histological lesions in the reproductive phygocytic tract. Persistent activity of infections neutrophils and reduce they are chemotactic responsible and for inflammatory damage leading to impaired fertility. In absence of accurate counts of the bacteria present in the uterus at the time of service and during the subsequent oestrus cycle, it is impossible to determine whether pathogenic bacteria cause the infertility by injuring the gametes or developing concepts. Pathogen gain access to reproductive tract due to breeding, parturition, veterinary examinations, retained placenta. Relaxed vulva, dilated vagina and open cervix facilitates the entry of pathogens in reproductive tract and hence uterine involution turns septic rather than aseptic process. Deosi et al. (1992) reported that majority of pathogen of endogenous origin establishment of may post not partum contribute metritis but substantially is only towards because of environmental contaminants. Pathogenic organisms cause uterine inflammation, denudation of its mucosa and increase its secretions, thus altering uterine environment. Bacterial invaders colonize the non involuted uterus where toxin produced by them is absorbed, which leads to septicemia and pyaemia. Tissue damage, a prominent feature of many anaerobic infections, results from the action of bacterial toxins and enzymes. The 31 endotoxin lipopolysaccharide (LPS), which is released from E.coli, can pass from the uterine lumen to the peripheral circulation and LPS concentrations are increased in cows with uterine infection. Prolapse of genitalia may be the cause of entry of severe external environmental infection due to long term exposure of genital organs. There are chances of urinary tract infection leading to straining and subsequent prolapse. Infection induces inflammation and denudation of mucosa of exposed organs resulting in tissue damage and release of highly irritating histaminic substances leading continuous straining (Sharma et al. 1977). Immunological aspects in non specific infections Ability of cows varies for resistance to continual uterine infections. Similarly ability to mount immunological and physiological defense mechanism also varies from cow to cow and herd to herd. There is decrease in immune function of animals after calving due to severe stress. Lewis (1997) reported that aberrant immune function predisposes for uterine infections in cows which indicates that methods regularizing immune function have the potential for prevention or treatment of uterine infections. Female reproductive tract, like respiratory and gastrointestinal tract, is continuously exposed to microbial organisms. Cervix is a major barrier to prevent microorganisms as lactoferrins inhibits microbial growth. However, immune defense mechanism is provided by phgocytes engulfing and killing antigen or microbes, B lymphocytes producing antibodies against microbes and T lymphocytes secreting cytokines to stimulate macrophages. Elimination of pathogens normally through reproductive tract is through leucocytic infiltration, persistent uterine 32 contractions, sloughing of caruncular tissue and secretions of uterine lochia as reported by Arthur et al¸ (1989). Uterine humoral immunity is expressed through secretions of immunoglobulins. Immunoglobulin concentration in uterine secretions reflects both the extent of the endometrial inflammatory process in the face of microbial challenge and its chances of cervical recovery (Aknarzov, 1988). Uterine defense mechanism remains inadequate during the progesterone dominated phase because low pH creates intrauterine environment favourable for bacterial infection (Gunnick, 1973). Innate immunity is the first defense to infection and is physiologically ancient than adaptive to fight against invading microbes through cellular components like lymphocytes, monocytes, neutrophills, macrophages, natural killer cells and molecular components like mucin, defensin, serpins, lectiferrins, interleukins, cystatinsetc. Possible application of immunomodulators and or cytokines for increasing uterine defense in case of microbial invasions and to monitor embryonic and fetal developments is currently under investigation (Yadav and Agrawal, 2002). Halls et al. (2002) has accounted host defense peptides and proteins expressed in the male reproductive tract. Modulation of reproduction functions through immunological approach is a recent concept and immnomodulators can be considered as alternatives therapeutic agents in clinical and sub-clinical infections. Immunization by antigens and by raising antibiotics against reproductive hormones has been used to develop experimental method for fundamental research in reproduction and to explore potential for augmenting reproductive efficiency in livestock. 33 Diagnosis of non specific infections Appearance of cervical mucus, vaginal discharge is usually considered as detection of reproductive tract infections. Character and odor of vaginal mucus can be scored for diagnosis of genital infections. Intrauterine oxygen reductase potential (Eh) and PH can be used to assess the degree of bacterial contamination of the uterine lumen. Contaminating bacteria are practically always present in the external genitalia. The most careful sampling procedure and especially designed sampling instruments can not eliminate contaminating samples entirely, which causes insurmountable difficulties. Zamjanis, R. (1970) reported that the number of micro-organisms or the titer of antibodies present in the genital tract varies greatly in certain infection diseases. In fact pathogens may disappear entirely from reproductive tract after being present for a certain period of time and negative results then may be due to sampling without selection of proper animals and the proper time. Cultural sensitivity tests give inconsistent results of uterine, vaginal and cervical swabs on diagnostic lines. Thus bacterial culture of cervical mucus could aid in diagnosis of infectious infertility but it should be interpretated in the light of clinical diagnosis and breeding history of the animal (Sharda and Krishnan, 2001). Isolation of bacteria from genital tract in absence of any clinical deformity or change in genital organs suggests role of bacteria in infertility. However, subclinical infection is difficult to diagnose and it is also time consuming, hence empirical treatments are undertaken by the field vets for infections of genital tract. 34 Petit et al (2008) reported that the percentage of positive vaginal swabs did not differ between pregnant and non-pregnant animals. In the genital tract, the percentage of swabs positive for normal mucosal bacteria decreased from caudally to cranially. Pathogenic bacteria were found more often in cervical than in vaginal swabs. Bacteria on the vaginal and cervical mucosa in cattle involve a wide range of species and their colonization of the mucosa rather than infection without endometritis or vaginitis in animals has to be assumed. Non specific infectious affections in male animals Various types of male infertility conditions have been reported in the literature like balanitis, posthitis, seminal vesiculits, prostatitis, urethral inflammation, testicular degeneration, orchitis, epididymitis, ampulitis etc. Under practical conditions, it is not possible to produce semen free from microorganisms as contamination with few non pathogenic organisms is unavoidable (Binda et al. 1994). Saprophytic flora of prepuce contaminates semen of healthy bulls at the time of ejaculation. Many microbes producing systemic infections but which are not specific to reproductive tract can be encountered in semen. Prepucial microflora of bull and vaginal microflora of cow gets transferred to uterus during insemination. Bacterial counts in semen vary with age, breed, season and prepucial washing. The principal part of contamination of semen consists of saprophytic or opportunistic organisms from prepuce and upper parts of genital organs. It is advisable to wash prepuce before semen collection with mild disinfectant lotion vary regularly to reduce bacterial load. Mean microbiological load was increased by four fold in second ejaculate (Gangadhar, et al., 1986), hence use same artificial vagina for two 35 successive ejaculates is not expected under clean semen production. The bacterial contamination of semen is major concern for semen production laboratories as the pathogens adversely affect the semen quality (Diemer et al.¸1986). Bacteria are contaminating approximately 50 per cent of frozen semen doses as reported by Weierzbowski et al. (1984). Artificial insemination is being widely practiced in the world and there is constant exchange of semen doses on international front. To presume that semen can be consistently microbiologically sterile in respect of non genital, specific, opportunistic, vagrant pathogens, which are umbiguious in the environment of cattle, are reported to be naïve by Bhosarekar (2005). Fungal infections may develop during processing of semen for preservation. Fungal infections of prostate and other accessory sex glands may result in presence of fungal infections in semen, which reduces quality of semen. Quality of semen depends on microbial load present in the semen and international standards regarding microbial counts are much less as compared to Indian standards. A permissible count of 500 bacteria per dose of semen straw is the international standard. Accordingly, it is recommended that prepucial washings (maximum 50,000 count) and neat semen (maximum 1000 to 5000 count) should carry lowest minimum bacterial load with no count for AI equipments and dilutors as they are to be used only after strict sterilization. Liquid nitrogen used for storage of frozen semen doses may act as vehicle for contaminant pathogens with variety of organisms and various. The same thus serves as source of infection to cows and buffalos during AI. Even fresh liquid nitrogen carry staphylococcus aureus. Certain bacterial contaminants acquire a level of resistance to antibiotics and they are able to survive at -190OC in liquid nitrogen (Ranold and Prabhakar, 2001). 36 Endocrine events and non specific infections Sub fertility due to non specific contamination involves perturbation of the hypothalamus, pituitary and ovary and pathogenic effects persist even after clinical resolution of disease. Pivotal role of progesterone in converting uterus resistant to susceptible for infection is important in female infertility. The uterine epithelium permeability to bacteria is reduced and the leucocytic response is delayed in progestational phase. Local production of antibody in the reproductive tract is also regulated by ovarian steroids. Absorption of bacterial components from uterus can prevent the follicular phase, LH surge and ovulation. There are also localized ovarian effects of high uterine bacterial growth density because fewer frontline dominant follicles are selected in ovary ipsilateral than contra lateral to the previously gravid horn. Uteri of animals during follicular and luteal phases differ significantly in their inflammatory responses to bacterial infection (Sadasiva Rao and Sheshagiri, 1999). Dairy animals are resistant to uterine infections when progesterone levels are minimal and they are susceptible when progesterone concentrations are increased. Although bacterial contamination can ieduce puerperal metritis soon after calving, when progesterone levels are basal, uterine infections do not usually develop till progesterone levels are increased. Uterine contamination following calving is common in 90 per cent animals but healthy animals are able to clear the bacteria of uterus in first 2 to 3 weeks of calving. In cows which develop uterine infection in later stages, bacteria reside in the uterus without proliferating into infection until luteal progesterone down regulates immune functions. First post partum dominant follicle is ovulated silently with perturbed 37 oestradiol secretion and subsequent formation of small corpus luteum, which secretes less progesterone. These reproductive events favor for delaying uterine infection. E. coli is a common cause of infertility involving the perturbation of the hypothalamus, pituitary and ovary in dairy cows. Infusion of E. coli LPS into the uterine lumen suppresses the pre-ovulatory luteinising hormone surge and disrupts ovulation in heifers. Even adequate concentration of treatment antibiotic in cyclic animals is not achieved in the subendometrial tissue, cervix and vagina during luteal phase (Gustafsson, 1984). Non specific infectious affection in female animal Types of female infertility problems related with nonspecific infections are aberrations in oestrus expression, gamete damage, failure of implantation, abortion, retained placeta, ovaro bursal adhesions, salpingitis, uterine infection, metritis, cervicitis, vaginitis, placentitis or cotyledonitis. Infection of the reproductive tract by non specific organisms is very common together with number of specific pathogens which selectively affect the reproductive system. Ball and Peters (2004) reported disturbances in reproductive activities due to non specific systemic infection with record of behavioral estrous inhibition due to mild systemic disease and embryo loss due to pyrexia as a resultant of some systemic infection. Induced endometritis significantly increases interoestus length when infection is introduced during luteal phase. Sporadic infections due to non specific pathogens also lead to abortions in pregnant animals. 38 Invading microflora of reproductive tract are important determinants of successful and speedy involution of uterus as well as subsequent fertility. Failure of immune system to control usual commonsal organisms principally leads to incomplete involution of uterus (Wheeler, 1994). Honnapagol, S.S. (2006) reported that uterine infections influence the time of first ovulation as animals with bacteriologically sterile uterus ovulate average 16 days after parturition, while infected ones are likely to ovulate average 31 days after parturition. Very few animals die due to uterine infections but many animals are culled regularly for their poor reproductive performance resulting due to non specific uterine infections as uterine infection can reduce milk yield, or in treatment cases contaminate milk. It is also important to know that because they are non specific uterine infections they are difficult to treat. A. pyogences either alone or with other bacteria is often associated with utrine infections. Khanna et al. (1992) reported 7.14 per cent incidence of mycoplasma infection in repeat breeder buffaloes and 6.25 per cent in cows due to cervicitis and endometratis. Heifers with incompletely developed cervicial rings and older cows with prolapsed cervical rings are prone to endometritis. It is difficult to identify the most significant microbes associated with endometritis because both pathogenic and non pathogenic organisms colonize in the bovine endometrium and many of them are fastidious (Studer, 1981). Vaginitis condition due to non specific infection is characterized by purulent vaginal discharge, red granular lesions and pain on palpation. The condition is also termed as “Dirty cow syndromes” and Haemophilus somnus infection was isolated in vaginitis by Last et al. (2001). 39 Control measures Sanitary and phytosanitary measures of WTO pertain to laws or regulations to protect livestock against exposure to micro organisms. No hand is microbiologically sterile and hence, strict attention to sanitation, peri-parturient hygiene, assisted calving may be the best defense for limiting non specific infection. Effective sexual health control programme with frequent bacterial examination in dairy animals need to be emphasized. In India, dairy animal farm construction is carried out without considering requirements of functional and productional aspects of the animals. The hygienic requirements are poorly considered during constructions. A hygienic environment in the animal house is essential for preservation of animal health and as an aid to control diseases. Super fire ways of restricting non specific infections needs to be followed for combating infertility or sub-fertility in dairy animals. Contamination primarily results from overcrowding and improper cleaning animal shades. Safe and sanitary disposal of liquid and solid wastes, quarantine of newly purchased and procured animals, isolation of suspected animals and foot bath for both human and animals are the most essential prerequisites for ideal and healthy farm. Thorough cleaning, complete draining, perfect drying of floorings of animals sheds and exposure to sunlight daily for six hours are the most ideal recommendations to keep the environmental infection away from the herd. Comfortable environment through proper ventilation for free removal of obnoxious odor is highly essential as the foul odor is the evil of all bad things in the herds. 40 Uterine infection depends on various factors like calving management, general sanitation, pathogenic organisms, endocrine factors, nutrition, milk production and environmental stress. But proper stages of oestrus, well built and body condition of animal, high level of intrinsic immunity, strong resistance to contaminants and proper defense mechanism will resists ascending infection in all probabilities. The uterine infections can be controlled by antibiotics, anti-septics, Hormones or immuno-modulatory treatments. Antimicrobials can cure bacterial, yeast, chlamydial, rikettesial and certain protozoal infection but not the viral or parasitic diseases. Objective of treatment in uterine infections is to stimulate the uterine muscles through higher vascularity either by increasing blood supply or to create healthy and sterile environment by checking uterine infection. Indiscriminate use of higher antibiotics has not only created most vital and serious problem of drug resistance but also has lead to flaring up of mycotic and anaerobic infections. Refractoriness to usual anti- biotic therapy boosts non specific infections. Anti-biotic can not be considered as total control measure in infections and no antibiotic can kill all known micro organisms. Drugs are therapeutically used to cure diseases and sub therapeutically to control outbreaks of diseases. However, it is pertinent to note that routine and prophylactic administration of intrauterine antibiotics to all cows with and without abnormal calving appears to enhance colonization of resistant bacteria within a herd and does not improve reproductive efficiency. Disappointing efficacy of intra uterine antimicrobials to combat infections is reported by various research workers and clinicians. Antimicrobial therapy also necessitates compulsory disposal of milk. 41 Emergence of anti-biotic resistant bacteria in the environment is also a global problem. The difficulty in controlling genital infection is due to emergence of drug resistance and the pattern of drug resistance continuous to change in a particular area, depending upon various epidemiological factors. However, immunomodulators (Oyster glycogen, Lipopolysaccharide etc.) can be considered as choice of treatment. Chronic endometritis fails to respond even to immunomodulators as it is already exposed to several intra uterine treatments. Use of nistatin as an antifungal agent was recorded to be successful in reducing fungal infection in preserved semen by Sidhu et al. (1997). Lactoferin is known for optimal health, immunity and resistant to diseases. It can be used commercially as natural antibiotic, antiviral and antifungal agent. Lactoferrin is considered to be bacteriostatic (Devatkal, et al. , 2003). The natural antibiotics or hosts defense peptides, i.e. antimicrobial peptides, are the part of innate immune system and are considered as antibiotics of the grate future. Alternately, polyherbal ayurvedic formulations are to be recommended and proved bacteriostatic action of herbal drugs in rabbit pyometra (Dakshinkar, 1999). It is worth noting to consider that Chinese farms decreased uterine infections from 54.8 to 25.00 per cent in just 5 years with concurrent advantage of improved post partum fertility as reported by Liu – Chong LI (1996). The challenge for future is to design prevention and control programmes to reduce the incidence of non specific infectious infertility conditions and to understand how the immune and endocrine systems are integrated to optimize reproductive efficiency in dairy animals. 42 References Aknarzov,B. K. (1988) Study of humoral factors of local protective function in cows. Veterinariya 8: 41-43 Arthur, G.H., Noakes, D.E. and Pearson, H. (1989). Veterinary Pearson, H. (1996). Veterinary Reproduction and Obstetrics. 6th Edn. Arthur, G.H., Noakes, D.E. and Reproduction and Obstetrics. 7th Edn. pp 392. 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