Achievements in Animal Health Management in Independent India
BN Tripathi1, RP Singh2, AK Tiwari3, G Saikumar4, GVPPS Ravi Kumar4, Yash Pal5, BR Gulati5, BR Shome6, VP Singh7, Jyoti Misri1, Triveni Dutt4 and Ashok Kumar1
1Indian Council of Agricultural Research, New Delhi
2ICAR-Directorate of Foot & Mouth Disease, Bhubaneswar, Odisha 3ICAR-Central Avian Research Institute, Izatnagar, Uttar Pradesh 4ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 5ICAR-National Research Centre on Equines, Hisar, Haryana
6ICAR-National Institute of Veterinary Epidemiology and Disease Informatics, Bengaluru, Karnataka
7ICAR-National Institute of High Security Animal Diseases, Bhopal, Madhya Pradesh
Summary
Livestock and poultry diseases continue to adversely affect the productivity and production of animals. Keeping in view the importance of animal husbandry sector, several animal health and species-specific institutes were established under the umbrella of ICAR to improve the animal health management system in the country. Technological interventions for over seven decades have led to the eradication of three diseases, namely; rinderpest (2006), contagious bovine pleuropneumonia (2007) and African horse sickness (2014). It has been estimated that rinderpest eradication has resulted in increase of milk production by 4.66 % with a net present value of Rs.3,463 crores (34.63 billion) and a cost benefit ratio of 10:43. Foot-and-mouth disease (FMD) and Peste des Petits Ruminants (PPR) control programs undertaken using indigenous technologies during last two decades also contributed to enhanced production. In spite of the growing human population in the country, the per capita milk availability has increased from 130 g in 1950 to 406 g in 2019-20. Similarly, the per capita per annum egg availability has increased from 5 eggs in 1950 to 86 eggs in 2019-20. Intensification of animal production system to meet increased demand for foods of animal origin, faster international trade and travel, and globalization have led to higher risks of new emerging infections including zoonotic and transboundary diseases. Incursions of avian influenza, porcine reproductive and respiratory syndrome (PRRS), lumpy skin disease (LSD), and African swine fever (ASF) in the last two decades, are the examples. ICAR institutes have developed more than 25 vaccines and 40 diagnostic test/surveillance kits and several molecular methods for the diagnosis and control of livestock and poultry diseases in the country. Also, these institutes keep a strict vigil over 30 emerging, re-emerging, and exotic diseases by systematic surveillance using indigenously developed diagnostic methods and help implementation of national animal disease control programmes.
Introduction
Livestock and poultry farming sustain rural livelihoods and contributes 25.6% to the agricultural GDP and 4.1% to the overall GDP of India (Bharadwaj et al. 2020). The growing demand for quality livestock products in human diet has promoted commercialization efforts, which in turn, has necessitated technological interventions for controlling animal diseases and efficient service delivery in the sector (Ahuja et al. 2008). Given the size and distribution of India’s livestock population, improvement of the livestock health and production presents a significant opportunity to enhance rural income and accelerate the pace of poverty alleviation. Animal diseases like FMD, LSD, haemorrhagic septicaemia (HS), brucellosis in cattle/buffalo, PPR, goatpox, sheeppox, contagious caprine pleuropneumonia (CCPP) in goat/sheep, classical swine fever (CSF), and African swine fever (ASF) in pig, Avian influenza (AI), Newcastle disease (ND) and other infectious diseases in poultry birds (Yadav et al. 2016) continue to inflict economic losses on livestock and poultry producers with trade implications.
Owing to the technological interventions of ICAR institutes, the livestock and poultry production and productivity have increased considerably. However, despite improved housing facilities, balanced nutrition, and disease control programmes including strict bio-security protocols, India is witnessing repeated occurrences of emerging, re-emerging, and transboundary diseases. The Indian poultry industry is facing major challenges due to many diseases, including complex chronic respiratory disease (CRD), Marek’s disease (MD), AI, ND, etc. CRD, caused by Mycoplasma gallisepticum and further complicated by avian pathogenic Escherichia coli (APEC) is a serious impediment to potential broiler production in the country. Post-mortem based diagnosis and treatment is often not accurate and effective since mixed infections are becoming dominant and posing difficulty for differential diagnosis. The porous international border across some of the North eastern states present opportunities for spread of many infectious diseases from neighboring China, Bangladesh, Bhutan, and Myanmar. This necessitates immediate attention through systematic surveillance, development of vaccines, and diagnostics for onsite use along with strict border control.
Initiatives to strengthen livestock and poultry health management in India
The ICAR-Indian Veterinary Research Institute (ICAR-IVRI) was initially established as the Imperial Bacteriological Laboratory in 1889 at Pune for conducting research for the protection of Indian livestock wealth from the dreaded diseases. The laboratory was shifted to Mukteswar in 1893, and later in 1913 it was expanded to Izatnagar, Bareilly to start large scale production of veterinary biologicals. In view of the main focus on production of veterinary biologics, the institute was renamed as Imperial Serum Institute in 1930, but as the institute expanded to conduct research on all contemporary disciplines of veterinary and animal sciences, it was renamed again as Imperial Veterinary Research Institute (IVRI) in 1936. Post-independence, this institute was finally renamed as Indian Veterinary Research Institute (IVRI). The most significant contributions of the institute in the field of vaccine development and production are depicted in Fig. 1.
Fig. 1. Important veterinary vaccines developed in the post-independence era
Over a period of time, two animal health institutes, namely ICAR- Directorate of FMD (DFMD) and ICAR-National Institute of High Security Animal Diseases (NIHSAD) were established from IVRI. Both these institutes are now reference centers for FMD and Avian Influenza as recognized by FAO and OIE, respectively. Research need for FMD was realized at the 11th International Veterinary Congress in 1930 and later in 1943 ICAR initiated an ad-hoc scheme entitled “Vaccination of Indian cattle against foot-and-mouth disease” at IVRI, Mukteswar. An “All India Co-ordinated Research Project (AICRP) for FMD virus serotyping” was initiated in 1968 with a central FMD laboratory at Mukteswar and three Regional Centers located at Hisar, Hyderabad, and Calcutta. During the year 2001, AICRP on FMD was upgraded to Project Directorate on FMD (PDFMD) with a network of 23 laboratories located all over the country. In the year 2015, PDFMD was renamed DFMD with 27 regional and collaborating centers. ICAR-DFMD is a member of the Global FAO/OIE network of FMD reference laboratories. The institute also functions as the FAO-FMD Reference Center and SAARC Regional Leading Diagnostic Laboratory for FMD. It is a member of GFRA (Global FMD Research Alliance). The state-of-the-art FMD research centre (ICFMD) with high containment laboratory facility established by ICAR at Bhubaneswar meets the major requirement of FMDCP as stipulated by OIE/FAO. High Security Animal Diseases Laboratory (HSADL), a regional center under IVRI, Izatnagar was upgraded to an independent national institute and was named ICAR-NIHSAD, Bhopal in 2014 with the main objective to deal with emerging exotic animal diseases and bio-risk management. The institute has Biosafety Level 3+ (BSL-3+) bio-containment facility available for research on exotic and high-risk pathogens of livestock and poultry in India. As a national referral laboratory, ICAR-NIHSAD is networking with all the 6 Regional Disease Diagnostic Laboratories (RDDLs) and provides diagnostic services for 30 exotic and emerging diseases. As an OIE reference laboratory for avian influenza, ICAR-NIHSAD provides diagnostic services to SAARC countries also.
Realizing the significance of disease epidemiology and informatics in disease control and eradication, ICAR upgraded the AICRP on ADMAS to PD-ADMAS in 2000 and subsequently to ICAR-National Institute of Veterinary Epidemiology and Disease Informatics (NIVEDI) in 2013. Among the three phases of rinderpest (RP) eradication, the phase of disease surveillance and monitoring was spearheaded by ICAR-NIVEDI by providing nationwide RP surveillance and monitoring plan, implementation strategies, and screening of serum samples. ICAR-NIVEDI is coordinating with other institutes and developed the national disease surveillance and monitoring plan for FMD, brucellosis, PPR, and CSF. The diagnostic kits developed by the institute are being used for sero- monitoring under brucellosis and CSF control programs.
Considering the importance of equine species, ICAR established National Research Centre on Equines (NRCE) in 1985 at Hisar and its regional station in 1989 at Bikaner. ICAR-NRCE has developed diagnostics against various equine diseases, including equine herpesvirus 1 (EHV1) infection, equine infectious anaemia (EIA), Theileria equi, glanders and inactivated vaccines for EHV1 and equine influenza. ICAR-NRCE has contributed towards declaration of disease-free status for African horse sickness (AHS) by OIE, and control of EIA and equine influenza. The National Centre for Veterinary Type Cultures (NCVTC), established in 2005 at NRCE, Hisar is working through 18 network units spread throughout the country and is maintaining a total of 3403 accessioned microbes.
Animal health institutes of ICAR monitor the quality of veterinary biologics produced by the industry through a stringent testing policy and certify for further use in the field. ICAR institutes have a key role in national animal disease control programs of Govt. of India by providing nationwide sampling plans, diagnostic kits, and epidemiological inputs to ensure success of animal health programs implemented by State and Central governments. Presently, technologies developed by ICAR institutes are being used for the diagnosis and control of four diseases viz. FMD, brucellosis, PPR, and CSF. R&D activities in the field of stem cell biology and their application in regenerative medicine has opened new avenues for treating non-communicable animal diseases. These interventions for improving the livestock health will ensure livelihood and nutritional security of the country .
Success stories of disease eradication
Eradication of rinderpest and its impact
Rinderpest, also known as “Cattle Plague” was one of the most devastating diseases of cattle and buffalo during the 18th and 19th century. The disease was caused by rinderpest virus (RPV) belonging to the Morbillivirus group. RPV also affected sheep, goat, and pig populations. Owing to the symptoms and death pattern, rinderpest in India was called “Ponkani”, “MaanRog” or “Pashu Mahamari”. The first Indian report on rinderpest was documented in 1752 by “Hallen Commission”. Clinically the diseased animals had shooting diarrhoea due to inflammation in the digestive tract. Nasal and ocular discharges were common in diseased animals. The death and infection rates in newly exposed populations were very high and sometimes up to 95-100 % leading to huge economic losses. Before the 1950s, a large number of bovine deaths were reported every year due to rinderpest in the country. The incidence of disease decreased to some extent (100-150 thousand) after 1935, when Goat Tissue Vaccine (GTV) developed at IVRI Mukteswar in the year 1927 (Edwards, 1927) was applied in livestock population. Subsequently during the 1st plan period (1954), a mass vaccination campaign against the cattle plague in the name of National Rinderpest Eradication Programme (NREP) using GTV vaccine was launched. The cattle and buffalo above 6 months of age were vaccinated using GTV vaccine. Subsequently, a safer vaccine developed by Plowright and Ferris (1962) namely Tissue Culture Rinderpest (TCRP) vaccine was introduced in India during 1970s (Yadav et al. 2016).
The TCRP vaccine was safe for pregnant and lactating animals, and conferred life-long immunity. Calf hood vaccination was done at 6-12 months of age and it was also efficacious in cross breed cattle, sheep, goats and pigs. Therefore, this vaccine was widely adopted. Between 1956-1989, a total of 1300 million doses of vaccination against rinderpest were carried out. These interventions led to reduction in disease outbreaks in India; however, extensive surveillance was required to prove absence of disease as per OIE pathway of rinderpest eradication. The overall sero-conversion rates/herd immunity of more than 70% were achieved at the completion of the national vaccination campaign in 2000. This effort totally stopped the transmission cycle of rinderpest virus. The mAb-based rinderpest competitive ELISA kit developed by ICAR-IVRI, validated by FAO and recommended by OIE, was used during the final stage of rinderpest eradication. The sero-surveillance using indigenously developed competitive ELISA kit (Singh et al. 2000) was continued till the year 2004 in order to prove that no virus is circulating in the population. The effect of vaccination on rinderpest related disease incidence & mortality is shown in Table 1.
Table 1. Plan-wise incidence and mortality of rinderpest in India
| Plan | Period | Outbreaks | Mortality | Mortality per million bovines |
| First | Prior to 1955 | 8000 | 200000 | 980 |
| Second | 1956-61 | 4368 | 31915 | 157 |
| Third | 1961-66 | 791 | 8348 | 27 |
| Annual | 1966-69 | 774 | 6146 | 22 |
| Fourth | 1969-74 | 237 | 2638 | 12 |
| Fifth | 1974-79 | 124 | 1200 | 5 |
| Annual | 1979-80 | 120 | 1296 | 5 |
| Plan | Period | Outbreaks | Mortality | Mortality per million bovines |
| Sixth | 1980-85 | 142 | 1596 | 6 |
| Seventh | 1985-90 | 173 | 1649 | 6 |
| Annual | 1990-92 | 52 | 247 | 1 |
| Eighth | 1992-97 | 47 | 148 | 0.5 |
| Ninth | 1997-2000 | 0 | 0 | 0 |
Following OIE pathway for rinderpest eradication, India became free from rinderpest in the year 2004. OIE recognized the country free from rinderpest infection in the year 2006 and finally, global eradication of rinderpest was declared by FAO in the year 2011. To mark this stupendous achievement which ushered an era of agricultural revolution in the country, ICAR-IVRI installed a commemoration pillar at its Mukteswar campus on 2nd June, 2012. The pillar depicts important landmark achievements made possible through contributions of all national and international organizations (Fig. 2).
The successful implementation of rinderpest eradication programme has yielded major economic benefits which enhanced food and nutrition security. It was observed that investment made in NREP launched in 1954 yielded net benefit and significant increase in growth rates in milk production. National Programme on Rinderpest Eradication (NPRE) was launched in 1991-92, to
Fig. 2. Memorial at IVRI Mukteswar to commemorate rinderpest eradication
eradicate rinderpest following the pathway prescribed by World Organization of Animal Health/ OIE. The benefits of NPRE can be appreciated from the increased access of India’s bovine meat and milk in the international market (Rich et al. 2012). Indeed, Indian exports of bovine milk picked up substantially around 1992-94 and simultaneously India’s dependence on milk imports also reduced drastically at the same time (Bardhan 2007). These developments ultimately contributed enormously to White Revolution and enhanced per capita availability of milk in the country. The growth rate and economic benefits during rinderpest control and eradication is depicted in Table 2. As against the huge economic benefits accrued from rinderpest eradication in India, the expenditure on rinderpest vaccine research and production was estimated to be Rs. 47,856 million (1063.46 million US$) over a period of 92 years from 1913 to 2005 (Yadav 2011). The FAO estimated that India gained additional food production valuing 289 billion US dollars between 1965-1998 (Yadav et al. 2016).
Table 2. Growth rate and economic benefits during rinderpest control and eradication
| Parameters | Growth rate/benefits |
| 1.a. Growth rates in milk production (1961 to 1983)
1.b. Growth rates in milk production (1983 to 2016) |
3.11% per annum |
| 4.66% per annum | |
| 2a. Net benefit during NREP (1956/57)
2b. Net benefit during NPRE (1994/95) |
Rs. 1576 million |
| Rs. 6933 million | |
| 3. Net benefits accrued to the nation during 1992/93 to 2015/16 | Rs. 4619 million per annum |
| 4. Net Present Value of rinderpest eradication | Rs. 34.63 billion |
| 5. Benefit Cost Ratio of NPRE (BCR) | 4.37 |
There have also been several indirect benefits of rinderpest eradication, which include capacity building in terms of human resource development, development of disease surveillance and diagnostic infrastructure, acquisition of vehicles and equipment, establishment of epidemiology units, creation of a wide network of veterinary departments and research institutes. Lessons learnt from rinderpest eradication are invaluable for achieving the goal of PPR control and eradication. The important milestones on the road to eradication of rinderpest and contagious bovine pleuropneumonia are given in Table 3.
Table 3. Milestone achievements of rinderpest and CBPP eradication post-independence
| Year | Milestones |
| 1954 | National Rinderpest Eradication Programme (NREP) launched. |
| 1967 | Large scale production of tissue culture rinderpest vaccine. |
| 1983 | National task force on rinderpest constituted. |
| 1987 | Establishment of AICRP on ADMAS at Bengaluru for rinderpest sero-surveillance. |
| 1992 | National Project on Rinderpest Eradication (NPRE) launched. |
| 2000 | Development of FAO/OIE validated rinderpest competitive ELISA kit. |
| 2003 | Provisional freedom from CBPP. |
| 2006 | OIE recognized India free from rinderpest infection. |
| 2007 | India declared CBPP free by the OIE. |
| 2011 | FAO Gold Medal for outstanding contribution to global rinderpest eradication program. |
| 2011 | Global freedom from rinderpest. |
| 2012 | Global Rinderpest Eradication Memorial installed at IVRI, Mukteswar. |
Eradication of contagious bovine pleuro-pneumonia
Contagious bovine pleuropneumonia (CBPP) was an insidious pneumonic disease of cattle and water buffalo referred to as lung sickness. It was caused by Mycoplasma mycoides subsp. mycoides (M. mycoides). Clinically, CBPP was manifested by anorexia, fever, and respiratory symptoms such as dyspnoea, cough, and nasal discharges. Eradication of CBPP was linked with the NPRE activities. The objective of the scheme was to strengthen the veterinary services to eradicate rinderpest and CBPP and to obtain freedom from rinderpest and CBPP following the pathway prescribed by OIE, Paris. The country was provisionally free from CBPP in October 2003. The eradication programme for CBPP was initiated in 8 districts of Assam and the dossier for CBPP eradication was submitted to OIE. India was declared CBPP free by the OIE in 2007. Now, it is important that the country’s freedom status against CBPP is maintained as per OIE requirements. The states and union territory governments are required to carry out physical surveillance up to the village level to maintain the freedom status of CBPP and to undertake surveillance of other animal diseases in the country on a routine basis.
Eradication of African horse sickness
African horse sickness (AHS) is a serious fatal disease of horses, mules, and donkeys. It is caused by a virus of the Genus Orbivirus belonging to the family Reoviridae. The virus is spread by infected insects (biting midges) and causes fever, cardiac and respiratory (breathing) problems with sudden death in affected animals. In India, the first case was reported in Cavalry of Army in April 1960. From 1960 to 1963, 22,977 horses got affected and 20,822 died (90.6% mortality). Initially, AHS7 virus strain vaccine was used to control the disease; however, this failed to provide sufficient protection. Later, indigenous mouse adopted strain 9 monovalent vaccine followed by AHS-9 chick embryo origin vaccine was developed to control the disease. The last report of AHS was in 1963 (Kumar 1976). In the 82nd General Session of OIE, India was declared free from AHS. Thus, IVRI played a lead role in eradication of AHS by contributing towards virus isolation, characterization, and development of vaccines and diagnostics.
Control of cattle and buffalo diseases
In addition to the eradicated diseases, several other livestock diseases are being monitored and controlled through different interventions.
Control of foot and mouth disease and its impact
Foot and mouth disease is caused by an Aphthovirus of the family Picornaviridae. Seven serotypes (O, A, Asia-1, C, SAT1, SAT2 & SAT3) are present globally. Out of the known serotypes, four serotypes viz., O, A, C, and Asia 1 were reported in India, before 1995. Now 3 serotypes (O, A, and Asia1) of the FMD virus are circulating in livestock in the country. Serotype C has not been recorded in India from 1995 onwards. The FMD affected large ruminants exhibit high fever, excessive frothy salivation, vesicles in the mouth especially on the tongue, teats, and inter-digital space, and decrease in milk yield due to reduced feed intake. Effective vaccines with short lived immunity are available; however, vaccines with long term immunity are needed. Indigenously developed state-of-the-art diagnostics are available for laboratory and field applications. Being a transboundary animal disease, technical know-how for FMD needs to be extended to SAARC member countries, which requires strong linkages with FAO and OIE.
The research on FMD vaccine started at IVRI, Mukteswar in 1943 with an ICAR sponsored Ad-hoc scheme “Vaccination of Indian cattle against FMD”. Crystal violet vaccine was prepared in the initial stages (1946-52), followed by goat kidney primary tissue culture methodology in 1964-65 and BHK-21 monolayer during 1972-77 at Mukteswar. With the increasing need for FMD vaccines, the Bengaluru campus of IVRI was established in 1972 for large-scale vaccine production using suspension cell culture. From 1971 to 1990, techniques for FMD virus typing, micro-complement fixation test, and microneutralization tests for subtyping were developed. ICAR driven FMD research and infrastructure development contributed immensely to FMD control efforts (Table 4).
Table 4. Milestones in foot and mouth disease control
| Year | Milestones |
| 1946-52 | FMD crystal violet tongue vaccine developed and updated. |
| 1968 | All India Co-ordinated Research Project (AICRP) for FMD virus typing launched. |
| 1971 | AICRP for Epidemiological studies on foot-and-mouth disease initiated. |
| 1972 | Bengaluru campus of IVRI established for large scale FMD vaccine production. |
| 1976 | Large-scale production of FMD vaccine started at Bengaluru campus ICAR-IVRI. |
| 1995 | Virus serotyping ELISA developed for FMD. |
| 2000 | AICRP on FMD upgraded to Project Directorate on FMD (PD-FMD). |
| 2003 | Liquid Phase Blocking ELISA (LPBE) for FMD developed. |
| 2003 | Uniform vaccine strain support to industry started. |
| 2004 | Multiplex PCR (mPCR) for FMD virus detection developed. |
| 2007 | PD-FMD became constituent laboratory of OIE/FAO FMD reference laboratories net- work. |
| 2008 | PD-FMD recognized as “FAO reference centre for FMD for south asia”. |
| 2009 | PD-FMD became Member Laboratory of Global FMD Research Alliance (GFRA). |
| 2009 | Recombinant non-structural protein (3AB3) based ELISA test developed for differen- tiation of FMD infected from vaccinated animals (DIVA). |
| 2009 | Foundation stone laid for International Centre for FMD (ICFMD), Bhubaneswar. |
| 2010 | PD-FMD became SAARC regional leading diagnostic laboratory of FAO. |
| 2013 | FMD lateral flow test and ELISA using recombinant antigen developed. |
| 2015 | PD-FMD upgraded to ICAR-Directorate of FMD (ICAR-DFMD) |
| 2016 | Solid Phase Competitive ELISA (SPCE) developed by ICAR-DFMD. |
| 2017 | Inauguration of International Centre for FMD, Bhubaneswar. |
| 2021 | ICAR-DFMD recognized as “FAO Reference Centre for FMD”. |
The FMD Control programme (FMDCP) was launched in 2003-04 in selected districts of India and expanded progressively to cover the entire country in 2017. A large number of FMD outbreaks were encountered prior to FMDCP. The number of FMD outbreaks/incidences came down by almost 60% (781 from 1911), in 2006/07 and in 2008/09 the number of incidences came down to 245, further ~70% drop in the incidences of the disease. The progressive drop in the incidences of FMD is attributed to the herd immunity. A trend of reduction in FMD outbreaks since implementation of FMDCP in 2003-04 is shown in Fig. 3. Through the application of the indigenous diagnostic kits India saved revenue worth Rs. 531 crores during last one decade.
Fig. 3. Reducing trend of FMD outbreaks in India after implementation of control programme
Socio-economic impact of FMD vaccines and diagnostic research
To evaluate the benefits of control measures on the production of affected animals, it is necessary to determine the variable effects of FMD including mortality rate (usually very low), reduction in milk production (usually significant), infertility, abortion, delays in attainment of slaughter weight, reduced food conversion efficiency in animals and lameness in bovine draught animals. It was projected that the state of Andhra Pradesh would stand to lose Rs. 1147 crore only on account of direct impacts, if there were no vaccination programme against FMD. The country would incur a total direct loss of Rs. 15575 crore. According to one study, the annual total economic loss due to FMD in India ranges from 12,000 crore to 14,000 crore (Singh et al. 2013). Another study in India estimated the benefit-cost ratio (BCR) to be between 5:1 and 8:1 (James and Ellis, 1978). Overall, existing evidence on BCR analysis of FMD vaccination in India and other endemic countries with comparable socio-economic status as that of India, strongly favours implementation of vaccination-based control of FMD for the benefit of livestock owners. There has been increase in milk yield and meat production in India over a period of time. Decline in incidences of FMD might have contributed to increase in milk yield and meat production in the country.
Control of brucellosis and its impact
Brucellosis is a bacterial disease caused by various Brucella species, which mainly infect cattle, swine, goats, sheep and dogs. Humans generally acquire the disease through direct contact with infected animals, by eating or drinking contaminated animal products. The seroprevalence of brucellosis in humans varies from place to place and also depends on the type of test method applied in the study. In a study conducted in and around Ludhiana, it was found to 26.6% using Standard Tube Agglutination Test (STAT) with a titre range between 80 and 1,280 IU/ml (Gamechu and Gill, 2011). For effective control of brucellosis, calf hood vaccination programme has been initiated under National Animal Disease Control Programme (NADCP). Reliable and verifiable diagnostics to support these efforts are available. This requires very strong linkages and awareness among stakeholders. IVRI provided diagnostic reagents for brucellosis at national level through production and supply of RBPT antigen, Milk Ring Test (MRT) antigen and SAT antigen. Recently, two indirect ELISA diagnostic kits have been developed one each at NIVEDI and IVRI. These diagnostic kits and the marker vaccine developed at IVRI (Brucella abortus S19∆ per vaccine) promise effective sero-monitoring and disease control.
The annual economic losses due to brucellosis in different livestock species in India were estimated on the basis of secondary seroprevalence data collected from published peer- reviewed literature and government reports. Meta-analysis was carried out to arrive at the pooled prevalence of bovine brucellosis. Various components of losses included in the study were reproductive losses (due to abortions and increased infertility), production losses, mortality losses in aborted animals and draught power losses. Simple mathematical models were developed to estimate the component-wise losses, which arrived at Rs. 9212 crores (Bardhan et al. 2020b). On account of the possibility of variation and uncertainty in various epidemiological and economic parameters, a sensitivity analysis was also carried out by considering worst-case and best-case scenarios. The benefit-cost ratio of brucellosis control through vaccination, under different scenarios, implied economic feasibility of vaccination.
Control of haemorrhagic septicemia and its impact
Haemorrhagic septicemia (HS) is an acute and often fatal disease of cattle and buffaloes, caused by Pasteurella multocida serotype B:2. The disease is characterized by high fever with concurrent shivering followed by profuse salivation, lachrymation, nasal discharge, and a sharp drop in milk yield. Pasteurella multocida, the cause of HS was first discovered by Perroncito in 1878 and the organism was isolated by Louis Pasteur in 1880. The work in India for control of HS was started at IVRI, as early as 1895 with the production of anti-HS serum. Due to the high cost involved in the production of serum and short-lived immunity, serum production was discontinued after the development of improved HS vaccines. Due to the devastating effect of the disease on livestock, the ICAR initiated the All India Network Programme on HS in the year 2000 to develop and improve vaccines against HS. A low volume saponified vaccine was developed to alleviate the problem of syringibility and swelling at the injection site in animals.
The economic loss due to HS in bovines was worked out as the sum of mortality loss, direct milk yield loss, losses due to increased abortions, drought power loss, cost of treatment, and extra labour costs (sample size 10,839 dairy animals). Simple mathematical models were developed for computing component-wise losses due to the disease. The economic loss per animal due to HS in India was estimated to be Rs.11,904; Rs.13,044 and Rs.20,296 in the case of indigenous and crossbred cattle and buffaloes, respectively. The share of buffaloes in the total economic loss was highest (55%), followed by indigenous (28%) and crossbred (16.5%) cattle. In view of the uncertainties associated with the epidemiological and economic parameters, stochastic modeling was used to estimate the economic impact of HS. The estimated annual economic loss due to HS in India was about Rs. 12758 crores (Bardhan et al. 2020a).
Control of infectious bovine rhinotracheitis (IBR)
IBR is a highly contagious disease caused by bovine herpesvirus 1 (BHV1). Apart from respiratory disease, which may lead to bovine respiratory disease complex or shipping fever after secondary bacterial infection, other clinical syndromes are infectious pustular vulvovaginitis (IPV) or infectious pustular balanoposthitis (IPB), abortion, conjunctivitis, infertility, arthritis, mastitis, and enteritis. During the past two decades, various types of vaccines developed include inactivated, live modified, subunit, DNA vaccines, and gene deleted marker vaccines. The gD-based subunit vaccine is considered most efficacious in reducing the clinical disease after they are combined with effective adjuvants like chitosan and CpG oligonucleotides. However, none of the presently available vaccines can prevent the establishment of latency of wild-type virus infection. Because of this, countries with vaccination programmes aiming at eradication were unsuccessful while the countries with programs of culling seropositive animals could achieve disease free status. The milestones for other cattle and buffalo disease control are given in Table 5.
Table 5. Achievement milestones in other diseases of cattle and buffalo for health improvement
| Year | Milestone |
| 1951 | Anthrax spore vaccine developed. |
| 1956 | Brucella abortus strain19 vaccine developed. |
| 1956-58 | Haemorrhagic septicaemia adjuvant vaccine developed. |
| 1979 | Bovine Theileria schizont vaccine developed. |
| 2001 | ELISA diagnostic kit for IBR and bovine brucellosis developed. |
| 2005 | NADRES was developed through “Weather based animal disease forecast (WB_ ADF)” and “Animal health information system through disease monitoring and surveillance (AHIS_DMS)” projects. |
| 2008 | AB ELISA for detection of IBR virus antibodies developed. |
| 2013 | Buffalo pox vaccine developed. |
| 2020 | Brucella abortus S19Δ per vaccine developed. |
Small ruminant health management and their impact
Small ruminant health management, related technology development and knowledge generation has been a priority activity of ICAR institutes. The important milestones in small ruminant health are shown below in Table 6.
Table 6. Achievement milestones in goat and sheep health improvement
| Year | Milestone |
| 1956-58 | Enterotoxaemia adjuvant vaccine developed. |
| 1973 | Irradiated sheep lung worm and multi-component clostridia vaccines developed. |
| 1986-87 | Sheeppox cell culture vaccine (RF strain) developed. |
| 2000 | PPR cell culture live vaccine developed. |
| 2001 | Monoclonal antibody-based PPR Competitive & Sandwich-ELISA kit developed. |
| 2006 | Live attenuated Goatpox vaccine developed. |
| 2012 | Indirect ELISA for sero-screening of brucellosis in sheep and goat developed. |
| 2013 | Cell culture attenuated live Orf vaccine developed. |
| 2015 | Indigenous Sheep pox vaccine (SRIN-38/00 strain) developed. |
| 2021 | ICAR-NIVEDI, PPR OIE Reference Laboratories Network (South Region). |
| 2021 | ICAR-IVRI Mukteswar, PPR OIE Reference Laboratories Network (North Region). |
| 2022 | Recombinant antigen based PPR competitive ELISA kit developed. |
| 2022 | Recombinant antigen polyclonal based PPR antigen capture ELISA kit developed. |
- Control of Peste des Petits Ruminants and its impact
Peste des petits ruminants (PPR), popularly known as “goat plague” is a contagious viral disease of small ruminants caused by a morbillivirus. The disease is found in several countries in Asia, the Middle East, and Africa. The control of disease is important from the point of view of livelihood security for millions of families involved in small ruminant husbandry. The important symptoms of PPR include fever, discharge from eyes and nostrils, conjunctivitis, gastroenteritis, and pneumonia. It was first described in Ivory Coast by Gargadennec and Lalanne in 1942, and in India from Tamil Nadu in the year 1987. The disease causes high economic losses in the country, as the infection rate in goat and sheep may reach up to 80-90%. The range of expected economic losses due to PPR was estimated to be between Rs. 4571 and Rs. 4683 crore/annum (Bardhan et al. 2017). By the years 2001 and 2002, the technologies with proven efficacy for vaccine and mAb-based diagnostic kits for antigen and antibody detection were developed.
The PPRV/Sungri/96 strain (Sreenivasa et al. 2000) is a vaccine virus strain used in India under mass PPR vaccination campaigns. This vaccine virus has been characterized extensively at antigenic and genomic levels (Singh and Bandyopadhayay 2015). The long-term immunity study indicated that the vaccine induces and maintains optimum virus-neutralizing antibodies for a long duration. Therefore, a single dose is sufficient for the protection of small ruminants. Large scale application of this vaccine to control the disease in India has been largely possible due to the Government of India’s supported PPR control programme and transfer of technology by ICAR to commercial manufacturers in the public and private sectors. The monoclonal antibody-based diagnostic kits namely sandwich-ELISA and competitive-ELISA have been used for diagnosis within the country for the last 20 years. The country is now self-sufficient in requirement of PPR vaccine and conventional/molecular diagnostics technologies including the recombinant antigen- based competitive ELISA kit (Balamurugan et al. 2021a), which may help during disease eradication programme. The trend of disease outbreak reduction during the last 15 years since the vaccines and diagnostics became available is shown in the maps below (Balamurugan et al. 2021b):
Fig. 4. District-wise outbreaks of Peste des Petits Ruminants over the years
District-wise cumulative outbreaks of PPR are considered one of the most important health constraints in rearing small ruminants. Various components of losses due to PPR in sheep and goats are mortality losses, reproductive failure, increased abortions, body weight loss, and treatment costs (Govindaraj et al. 2016). In addition, wool loss, increased inter-lambing period in case of sheep and milk loss and increased inter-kidding period in case of goats are other important losses (Bardhan et al. 2017). The benefit-cost of ‘Mass Vaccination Campaign’ in Chhattisgarh state reported benefit: cost ratio, net present value, and internal rate of return of 4.9:1, Rs. 342 crores and 146.6% under low incidence scenario; 12.4:1, Rs. 998 crores and 430.4% under medium incidence scenario and 13.5:1, Rs. 1096 crore and 430.4% under high incidence scenario respectively (Govindaraj et al. 2019). The change in total economic surplus due to vaccination, research, and delivery cost were projected from 1997 (the year of the start of the research project) to 2030 (by which 100% of the small ruminant population is to be vaccinated, as per OIE/ FAO specifications for disease eradication) after adjusting to the above adoption pattern. The benefits to society (economic surplus) and costs of the control programme were deflated using a suitable consumer price index to 2016 level. Using a long-run discount rate of 7.5%, the benefits were compared to research and delivery cost, and the net present value (NPV), internal rate of return (IRR), and benefit-cost ratio (BCR) were calculated. Using the economic surplus model, the change in total surplus as a result of mass vaccination of sheep and goats against PPR in India was found to be Rs.8,253 crore per annum with BCR and NPV of 123:1 and Rs. 480 crores, respectively (Bardhan et al. 2017). It is estimated that indigenous PPR diagnostics may have saved Rs.6.22 crore worth foreign exchange through import substitution (Singh et al. 2009), which may have increased several folds by now.
Sheeppox and goatpox
Sheeppox and goatpox diseases are caused by capripox viruses, all of which can infect sheep and goats. Modeling studies from the data collected in Maharashtra suggested that it would take about 6 years for a flock or herd to recover from an outbreak, with 30- 43% average annual losses in income, depending on flock type and the owner’s actions (Garner et al. 2000). An inactivated goatpox vaccine was developed at IVRI (Yadav et al. 1986) for field use till the development of a vero-cell culture-based attenuated vaccine for goatpox at ICAR-IVRI, Mukteswar during the year 2006. The lyophilized cell culture vaccine provided immunity for about 40 months (lifelong). This vaccine has also been used for the control of LSD in cattle as a strategy for emergency vaccination. For sheeppox, the RF strain of sheeppox vaccine, based on LT-32/Vero-9 cell culture was developed in 1986-87 .It induces protective antibodies for at least one year. The vaccine is safe for pregnant animals and young ones. Another Vero cell adapted sheeppox vaccine was also developed from an indigenous virus strain (SRIN-38/00) by ICAR-IVRI, in 2014. The scalability of the vaccine and downstream processing is simple and easy. The vaccine confers protection for up to 4 years.
Pig health management and their impact
Classical swine fever (CSF), is a contagious viral disease of domestic and wild pigs. It is caused by a virus of the genus Pestivirus of the family Flaviviridae. Animals with the acute disease die within 1-2 weeks. With low virulence strains, the only expression may be poor reproductive performance and the birth of piglets with neurologic defects such as congenital tremors. ICAR-NIVEDI has developed two diagnostic kits viz. (i) Indirect ELISA using recombinant antigen for detection of antibodies against CSFV in pigs (ii) CSFV Ag Check Kit for detection of antigen in clinical specimens. Vaccination can prevent the spread of the disease. A lapinized vaccine virus (Weybridge strain), was adapted to grow in cell line at ICAR-IVRI, Izatnagar. The cell culture vaccine is safe and potent. Each dose contains at least 100 PD50 and provides immunity for a year. Similarly, another vaccine strain (IVRI-CSF-BS) using an Indian isolate of CSFV has been developed and the technology transferred to industry.
Equine health management and their impact
Glanders
Glanders is a fatal infectious and notifiable zoonotic disease of equids caused by Burkholderia mallei. The long duration of therapy and unavailability of vaccine makes this pathogen formidable to control. The existing control policy dictates the identification and elimination of seropositive equines. The complement fixation test (CFT) is the OIE- prescribed serodiagnostic method for glanders; however, it produces false-positive or doubtful results with donkey and mule sera. ICAR-NRCE developed three recombinant protein-based ELISAs in 2012-13, which were validated in OIE reference laboratory on glanders, Germany during 2015-2017 and Hcp1 ELISA showed superior performance with 95.28% sensitivity and 99.56% specificity (Elschner et al. 2019). This ELISA has been extensively used for surveillance of glanders by State Disease Diagnostic Laboratories (SRDDLS) and Regional Disease Diagnostic Laboratories (RDDLs) of Govt. of India. More than 150,000 equines were tested by the Hcp1 ELISA (Singha et al. 2020). The ELISA has proved to be safe, rapid, inexpensive, accurate, and user friendly to adopt in a diagnostic laboratory with limited resources. The test has proved to be very useful in state- wide surveillance and control programmes of glanders and has been commercialized.
Equine influenza
Equine influenza (EI) is an OIE listed respiratory disease of horses, mules, ponies, donkeys, and zebra, caused by two strains of Influenza A virus viz. H7N7 and H3N8. Since 1980, no outbreak has been reported due to H7N7. Outbreaks due to H3N8 have been regularly reported from different parts of the world. The disease is highly contagious and spreads very fast through the aerosol route, and is characterized by fever, dry hacking cough, and watery nasal discharge, which later become mucopurulent. In India first report of influenza-like symptoms was from erstwhile Bombay in the Bombay Turf Club in 1964, where around 400 horses had an outbreak of coughing. Subsequently, two major epizootics have been reported. The first one was in 1987 when around 83,000 horses were infected mostly in northern Indian states (Uppal et al. 1989). The second epizootic occurred during 2008-09 in Katra (J&K) from where it spread to 14 States affecting thousands of animals and leading to huge economic losses to the stakeholders (Virmani et al. 2010). Phylogenetic analysis indicated clade 2 of the Florida sub-lineage of the H3N8 virus. ICAR-NRCE developed an inactivated low-cost vaccine using a virus from the outbreak of 1987. Subsequently, an updated inactivated equine influenza vaccine was developed during 2008-09, which was followed by a recombinant vaccine candidate through reverse genetic engineering. ICAR- NRCE has also developed several diagnostics including haemagglutination-inhibition (HI) assay for serological diagnosis, RT-PCR, qRT-PCR, monoclonal antibody-based sandwich ELISA, RT-PCR for subtyping of EI virus, and immunohistochemical diagnosis.
Equine herpesvirus infections
Equine herpesviruses (EHV1 and EHV4) are the most important pathogens that infect 80 to 90% of horses by two years of age, resulting in respiratory infection, characterized by fever, anorexia, nasal and ocular discharge. EHV1 causes upper respiratory tract infection in young horses at the time of weaning, abortion in pregnant mares, neonatal foal mortality, and neurological disorders. Abortion is economically most crippling outcome of EHV1 infection with 95% of EHV1 associated abortions occurring in the last four months of pregnancy. Abortions in pregnant mares range from 4-8% in organized horse breeding farms in India, which is the major cause of economic losses in equine industry. For timely diagnosis of EHV1, NRCE has developed various diagnostic assays including HERP kit in 2003, monoclonal antibody-based blocking ELISA kit in 2008 and recombinant protein- based ELISA for differentiation of EHV1 and EHV4 infection (NRCE Annual report 2016- 17 and 2017-18). These diagnostics have contributed towards timely diagnosis and control of disease in equine population. In addition, an effective inactivated vaccine ‘Equiperabort’ for control of abortions in pregnant mares has been developed (Singh et al. 2009a). This vaccine has been extensively tested in field trials and is now being used in organized equine farms, including Equine Breeding Studs of Indian Army.
Equine infectious anemia
Equine infectious anaemia (EIA) is a persistent viral infection of equids, caused by Lentivirus of Retroviridae family. The EIAV is mechanically transmitted from infected to susceptible equids by biting horse flies, deer flies and stable fly. Once a horse is infected with EIAV, it remains infected for rest of the life and a potential source of infection to other horses. There is neither effective vaccine nor treatment for this disease. It is one of the notifiable equine diseases, which entails implementation of strict control policy including elimination of the sero-positive equids. ICAR-NRCE has been regularly monitoring EIAV infection in the country. In India, first case of EIA was detected in race horses in Karnataka in 1987 (Uppal and Yadav 1989). Later, maximum number of EIA cases (n=186) were detected during 1987-1990 but thereafter, it was rarely reported. As there are no specific pathognomonic clinical signs in EIA, demonstration of EIAV specific antibody in the serum is required for confirmation of the infection. Agar gel immunodiffusion (AGID) test (Coggins test) is the OIE recommended test; however, it lacks sensitivity and may give false negative results. The test is also time-consuming & requires 48 to 72 hours. Therefore, ICAR-NRCE developed a recombinant p26 protein based indirect ELISA for EIA diagnosis (Singha et al. 2013) and the diagnostic technology has been commercialized.
Trypanosomiasis (Surra)
Trypanosomiasis, caused by Trypanosoma evansi, is an important disease of equines resulting in high morbidity and mortality. Clinical symptoms mainly are fever, anaemia, reduced milk yield, weight loss, lower work output, abortion, infertility, and in many cases, a deteriorating condition, that results in death. Total annual economic loss associated with surra in equines has been estimated to be Rs. 146.87 million (Rs. 61.45 to 293.92 million at a 95% confidence interval). ICAR-NRCE is currently monitoring the prevalence of T. evansi in equines in India, using antibody ELISA developed by the institute (Kumar et al. 2013a). A total of 20,609 equine serum samples from different states were tested up to March, 2021 and 1077 (5.22%) equids were detected positive. It has helped in risk assessment of trypanosomiasis in endemic areas. Diagnostic service has helped the animal owners in timely initiation of treatment and saving their precious animals as well as check further spread to in-contact healthy animals.
Equine piroplasmosis
Equine piroplasmosis, a tick-transmitted haemoprotozoan disease caused by intraerythrocytic protozoa Theileria equi and/or Babesia caballi is an economically important disease of equids. Equine babesiosis is transmitted by Ixodid tick species of genera, Hyalomma, Dermacentor, and Rhipicephalus. Diagnosis of most acute haemoparasitic infections is routinely done by microscopic examination of thick and thin smears. However, serodiagnosis following clinical or sub-clinical disease is a powerful tool in detecting and defining the prevalence of protozoan diseases. NRCE developed recombinant antigen- based ELISA kit in 2008 for the diagnosis of T. equi antibodies and testing of one sample costs Rs. 55 only, which is quite economical as compared to a commercial kit by VMRD, USA costing Rs. 610/- per sample (Kumar et al. 2013b and Kumar et al. 2015). Milestones registered in equine, porcine, canine, and camel health improvement are given in Table 7.
Table 7. Milestones in equine, porcine, canine and camel health improvement
| Year | Milestone |
| 1962 | Lapinized swine fever vaccine developed. |
| 1982 | Anti-Rabies BPL inactivated vaccine developed. |
| 1991 | Rabies cell culture vaccine (BHK-21) developed. |
| 1996 | Inactivated low cost vaccine for equine influenza developed. |
| 2003 | HERP kit for Equine herpesvirus infections developed. |
| 2007 | Development of cell culture vaccine for classical swine fever. |
| 2008 | Monoclonal antibody-based blocking ELISA kit against EHV1 developed. |
| 2008 | Recombinant antigen-based ELISA kit for diagnosis of T. equi antibodies developed. |
| 2008 | Recombinant protein-based ELISA for differentiation of EHV1 and EHV4 infection de- veloped. |
| 2008 | Inactivated vaccine Equiperabort’ against EHV1 developed. |
| Year | Milestone |
| 2014 | India declared free from African Horse Sickness by OIE. |
| 2014 | Vero cell culture attenuated live camelpox vaccine developed. |
| 2015 | Updated inactivated equine influenza vaccine developed. |
| 2019 | Glanders ELISA kit developed. |
| 2019 | Recombinant p26 protein based indirect ELISA for equine infectious anaemia developed. |
| 2020 | Live attenuated Classical Swine Fever cell culture vaccine using indigenous strain developed |
| 2021 | Trypanosoma evansi diagnostic kit developed. |
| 2021 | Inactivated Japanese encephalitis vaccine for pigs developed. |
| 2021 | Canine distemper indigenous vaccine developed. |
Poultry health management and their impact
Indian poultry is facing a major challenge in the control of diseases. Despite strict bio- security protocols, enhanced housing facilities, and caring nutrition, India is witnessing repeated occurrences of emerging and re-emerging diseases, especially in the organized sector. Now the most alarming concern is that the post-mortem based diagnosis and treatment is often confusing since mixed infections are becoming dominant and pose difficulty for differential diagnosis. Even vaccine strains are sometimes causing the disease in birds after vaccination. The north-eastern part of India shares a porous border with China, Bangladesh, Bhutan, and Myanmar adding high burden of infectious diseases to Indian poultry flocks. Pathogenic and emerging diseases namely avian influenza often causes heavy loss both in the domestic market and international trade. Respiratory disease complex and complicated chronic respiratory disease in poultry are other major challenging issues posed to the Indian poultry industry.
Avian influenza
Except for avian influenza, all poultry disease outbreaks are going almost unnoticed, hence remain underreported. Highly pathogenic avian influenza (HPAI) caused by the H5 subtype of Type A influenza virus has emerged as an economically most important disease with a significant impact on marginal and rural backyard poultry farmers (DAHD 2015). The outbreaks have occurred as epidemic waves during 2008-09 and thereafter established as sporadic occurrences. Frequent detection of H5N1 infections in north eastern and eastern regions (sharing borders with Nepal, Bangladesh, Bhutan, and Myanmar) indicates a regional cross-border problem with porous borders and illegal movement of poultry and poultry products contributing to the threat of potential endemic circulation within the region (Dhingra et al. 2014). Unlike other countries, India is continuously experiencing a frequent outbreak of the HPAI H5N1 strain (DAHD 2015). A survey report on poultry economics in West Bengal (Otte et al. 2008) indicated that economic impact through losses largely exceeds the monetary support provided by Govt. and therefore, there is little incentive for farmers to report infection, which compromises the efficiency of passive surveillance. It has been observed that the initial introduction of HPAI to a country is usually associated with long-distance transmission from infected areas through migratory birds (Newman et al. 2012). The massive border migration involvement in northeastern and eastern regions must be controlled with stringent border security measures. An amount of Rs. 26.44 crore has been paid from february 2006 to 23rd march 2020 as compensation to poultry farmers on account of culling due to avian influenza based on each outbreak. Series of outbreaks till 2009 cost the poultry industry a loss of Rs.30 crore. To date, 32 outbreaks occurred with two H5 serotypes that resulted in the death and culling of 4.36 and 87.24 lakh birds, respectively. Nevertheless, no such scientific and organized study has been conducted in India regarding the economic loss incurred due to the avian influenza outbreaks in poultry concerning each outbreak that happened between February 2006 to March 2020. India adopted stamping out protocol as the main strategy for the control of AI. ICAR-NIHSAD, Bhopal has made a tremendous effort in timely and accurate diagnosis of high and low pathogenic avian influenza in India, since the occurrence of the first outbreak in 2006 in the country. Some of the notable diagnostic achievements include development of avian influenza antibody detection ELISA kit, multiplex real-time RT-PCR kit for avian influenza A virus typing, and H5 and H9 subtyping and Lateral Flow Test for rapid detection of H5 avian influenza virus antigen in poultry. Important milestones in AI and other poultry diseases are mentioned in Table 8.
Table 8. Milestones in poultry health improvement
| Year | Milestone |
| 1945 | Ranikhet disease vaccine (R2B Mukteswar strain) developed. |
| 1946-47 | Fowlpox vaccine developed. |
| 1953 | Ranikhet disease vaccine (F strain) developed. |
| 1959-60 | Fowlpox vaccine (Egg adapted) developed. |
| 1999-2001 | Development of IBD vaccine. |
| 2000 | Preparedness for avian influenza diagnosis initiated. |
| 2001 | ICAR-NIHSAD recognized as national referral facility for avian influenza by
DAHD, Govt. of India. |
| 2009 | ICAR-NIHSAD recognized as international avian influenza reference laboratory
by OIE. |
| 2010 | Diagnostic services for avian influenza for Bhutan. |
| 2013 | Diagnostic services for avian influenza for Nepal. |
| 2016 | Development of reverse genetics based rgH5N2 DIVA marker vaccine for HPAI. |
| 2019 | Sub-viral particle based infectious bursal disease (IBD) vaccine. |
| 2021 | Inactivated vaccine for H9N2 virus developed. |
Newcastle disease
Newcastle disease virus (NDV) is prevalent worldwide and often spreads rapidly during epizootics in poultry, causing severe economic loss due to disease and, for countries that export poultry or poultry products due to trade embargoes. It spread rapidly in Asia and became panzootic within four decades (Alexander et al. 2012). ND has existed in India for the past 85 years and is also known as Ranikhet disease (RD) where the disease was first noticed and described. In India, almost every farmer carries out ND vaccination (Vegad 2014). The seroprevalence of ND may be as high as 83% in the country. It is the single greatest constraint limiting productivity and development throughout the developing world. Heterogeneity within strains of NDV may play a very important role in the maintenance and development of infection in village poultry populations. R2B (Mukteswar), a mesogenic vaccine strain of ND, is the popular vaccine strain used in the Indian subcontinent, especially in older birds (6–8 weeks old) with long-lasting immunity, but has proven to be pathogenic for young chicks. This vaccine strain had its origin by passaging one of the three Indian field isolates at IVRI, Mukteswar in 1945 and has been used as a vaccine candidate for booster immunization since then (Iyer and Hashmi 1945). This technology has been adopted by the biggest poultry vaccine manufacturers viz., Indovax Private Limited, Haryana; Hester Biosciences, Gujarat; Venkys Private Limited, Maharashtra, and billions of doses have been produced. A recombinant antigen-based ELISA kit has also been developed for active and passive sero-surveillance for Newcastle disease in commercial and backyard chickens.
Infectious bursal disease
Infectious bursal disease (IBD) seen in young domestic chickens is caused by infectious bursal disease virus (IBDV). Symptoms of the disease can include depression, watery diarrhoea, ruffled feathers, and dehydration. Morbidity is high, and mortality is usually low, but some very virulent strains are capable of causing 60% or higher mortality. Macroscopic and microscopic lesions in the cloacal bursa and molecular identification of the viral genome are used for the diagnosis of the disease. Very virulent pathotypes of IBDV emerged in 1992, resulting in huge economic losses to the poultry industry. Currently, IBDV is endemic and a serious problem for the poultry industry in India. Virulent, very virulent (vv), and classical forms of the disease are present in India with no reports on variant form of IBDV. Sequence alignment of these viruses with reported viruses of other countries revealed Indian IBDV field isolates to be 100% similar to very virulent Japanese (OKYM), European (UK661), and Bangladesh (BD3/99) IBD viruses at the amino acid level. Whereas they had 0.2- 0.9% divergence at the nucleotide level. Vaccination to induce maternal immunity in young chicks is initially used to control the disease. Vectored and live-attenuated vaccines can be used to induce active immunity in chicks as the maternal antibodies wane. At ICAR–IVRI, recombinant antigen-based sero-diagnostic assay for IBD has also been developed.
Other achievements on animal health management
Diagnostic pathology
Pathomorphological diagnosis has come a long way and plays an important role in the prognosis and diagnosis of various livestock and poultry diseases. At ICAR-IVRI, the fluorescent antibody test for diagnosis of Johne’s disease (JD) and the biological mouse inoculation test for rabies were applied for the first time. A modified Periodic Acid- Schiff reagent for routine staining and turpentine oil as a clearing agent were developed. For diagnosis of poultry diseases, the chicken embryo susceptibility test for avian encephalomyelitis in 1975, micro HI for ND and EDS-76, and MATSA test for Marek’s disease in 1984 were standardized. In 1986, Reovirus and vvIBDV were detected for the first time in the country. Inactivated, oil-based tissue culture and/or embryo origin vaccines against ND, IBD, Reo, EDS-76, IBH and DVH (Duck Viral Hepatitis), and combined inactivated vaccines like ND, EDS-76 and IBD were developed. In 1986, the COFAL kit for monitoring ALSV infection was developed. Avian leukosis virus subgroup “A” was recovered in 1991 for the first time in the country. Other diseases/conditions documented were: avian spirochetosis, avian aspergillosis, avian tuberculosis, mucormycosis, coccidioidomycosis (sheep and goats), protozoan/parasitic diseases, urea poisoning, HCN and nitrate/ nitrite poisoning and aflatoxicosis in poultry and cattle, and various tumors in animals and wildlife.
The reliability of PPD tuberculin/Johnin skin test was studied by examining pathological lesions at post-mortem. Experimental studies were conducted in goats to elucidate the pathogenesis of goiter and molybdenum-induced secondary hypocuprosis. From 1990 to 2010, extensive research was carried out on JD to develop and adapt various diagnostic methods such as bacterial culture, AGID, ELISA, PCR, RE analysis, and DNA probes. The experimental models (sheep, goat, rabbit, and mouse) for JD, entry of Mycobacterium avium sub sp. parartuberculosis (MAP) organisms via M cells and enterocytes and cytokine profile in sheep with pauci- and multibacillary pathology were established. On molecular typing, majority of Indian MAP isolates were found to be “bison” type and PFGE type 25. Pathogens associated with neonatal calf enteritis, viz., E. coli, rotavirus and coronavirus were also studied. The prevalence of Enzootic Bovine Haematuria (EBH) in Uttarakhand, levels of ptaquiloside and quercetin toxins in different ferns, and its pathology was studied. The cutaneous and teat warts and rumen/reticulum/urinary bladder mucosal growths showed involvement of bovine papillomavirus (BPV)-1 and -2 in cattle/ buffaloes and yaks, BPV-1 and -2 in equine sarcoids, BPV-1, -2, -5 and -10 and their combinations in cattle and buffaloes in EBH associated urinary bladder tumors.
Extensive research was carried out on classical swine fever (CSF) in pigs. Probe based RT-PCR assays were developed for detection and quantification of virus load in pigs with different clinical forms of CSF. The pathogenesis of CSF was studied using tissue based nucleic acid probes (DNA and RNA) and Indian isolates of CSFV were genotyped as 1.1 and 2.2. Swine influenza in pigs was confirmed for the first time in 2009, and the virus was shown to share close homology to the human H1N1 pandemic virus. Post-weaning multiple wasting syndrome (PMWS) caused by porcine circovirus-2 (PCV2) was reported for the first time in the country in 2005. PCV2 was genetically characterized as PCV2a and PCV2b and its recombinants. Other pig pathogens detected were porcine parvovirus, rotavirus, enterovirus, sapelovirus, Japanese encephalitis virus, Bordetella bronchiseptica and Streptococcus suis.
Parasitic diseases
The development of vaccines against lungworms of sheep and bovine tropical theileriosis were useful for control of these diseases. For lungworm control, a gamma radiation- attenuated D. filaria vaccine (Difil) was developed by IVRI in 1971. The vaccine constituted infective stage, L3 filarial larvae, radiation attenuated at 50 krad. The ‘Difil’ vaccination significantly reduced the incidence of lungworm infection in sheep in the temperate Himalayan region. A series of ectoparasitic management practices and an anti-tick vaccine were developed at IVRI for tick management. The advent of immunological and molecular techniques helped parasitologists initiate work on characterization of parasite antigens and their use in diagnosis of parasitic diseases. Serodiagnostic tests viz., IFAT, ELISA, dot- ELISA, s-ELISA, EITB, and LAT were developed and standardized for accurate diagnosis and studying seroepidemiology of trypanosomosis, babesiosis, theileriosis, toxoplasmosis, hydatidosis, cysticercosis, toxocariasis, ancylostomiasis, prepatent fasciolosis, and haemonchosis.
Surgical interventions
Veterinary surgery is one of the dynamic and enterprising areas of veterinary science research and practice. Different surgical techniques like castration, dehorning, hoof trimming, caesarean section, etc. have been part of the animal husbandry and management system. However, veterinary surgery has undergone a sea change in the past 2-3 decades with many advancements taking place in the areas of anesthesia and pain management, minimally invasive surgical techniques, and diagnostic imaging. At ICAR–IVRI, these techniques include – treatment of posterior paresis using stem cell therapy, treatment of a hernia using acellular biomaterial, tube cystostomy in goat and bullock, Epoxy-pin fixation for treatment of open fracture in calf, treatment of compound fracture in horse using circular fixator, interlocking nailing of tibia in cow, etc.
Stem cell biology
Stem cell biology is currently one of the most potential areas of biomedical research, which can revolutionize both medical and veterinary sciences. ICAR-IVRI has conducted research on stem cells for therapeutic application in livestock and pets. Emphasis on basic research is required before stem-cell-based therapies are widely used in veterinary sector. Embryonic/ induced pluripotent stem cells could be used as a reference model to understand important molecular signaling pathways which control cell fate decisions and organ differentiation.
The clinical use of stem cells in veterinary sciences is clearly in its early stages and various approaches are still being investigated. To preserve the germplasm of threatened/wildlife species, stem cell-based approaches could provide an opportunistic basis in the form of xenografting of testis tissue obtained quickly after the death of pre-pubertal animals.
Challenges and way forward
Development of improved vaccines (thermo tolerant vaccines, marker vaccines, etc.) and point of care diagnostics should be the thrust areas of research and development. For FMD, serotype-specific monoclonal antibody-based ELISA needs to be developed to replace polyclonal antibody-based ELISAs. There is also an urgent need for a cost-effective vaccine with long-term immunity for FMD control. Traditional methods for quality control of veterinary vaccines and drugs can be time taking and expensive, therefore there is an urgent need to develop alternate models that obviate the use of animals. Ethical use of experimental animals is also the compelling reason to develop alternate systems for quality control of veterinary vaccines and therapeutics. The emergence of Lumpy skin disease (LSD) in cattle and its rapid spread is a significant threat to cattle farming in India. Control of LSD mainly relies on early diagnosis along with restriction on movement of cattle in the affected areas, effective vector control measures, proper vaccination, and awareness of livestock farmers. ICAR institutes (IVRI and NRCE) are working on development of a candidate vaccine for LSD in cattle. Important diseases like brucellosis can be controlled to some extent by calf hood vaccinations, but, R & D for developing therapeutic strategy of the livestock is required. Similarly, some strategy for the control of Bovine tuberculosis is also need of the hour. Even with the proven animal health technologies, the major challenge lies with the implementation of disease control program in a diverse country like India, as the execution part rests with the state animal husbandry departments. CSF control in pigs is very important and a cell culture vaccine developed by IVRI is expected to contribute immensely to its control. But new diseases such as PRRS in 2013 and ASF in 2020 have entered the Indian pig population. These diseases cause heavy mortality and morbidity in swine. The complex nature of the PRRS and its genetic and antigenic variations pose a great challenge in the diagnosis and control of the disease. The management of farms and related biosecurity, diagnosis and surveillance, lack of public awareness, and concerns from across the border are the major challenges to control of PRRS and ASF in India. Avian influenza continues to be a serious concern because of its destabilizing effect on the poultry sector and public health risk. Country wide surveillance, early diagnosis of avian influenza along with subtype identification will help implementation of rapid and effective control measures to check further spread.
In the past two decades, a large number of new viral infections with severe life-threatening and economic consequences have emerged. Severe acute respiratory syndrome (SARS), Middle East Respiratory Syndrome (MERS), and SARS-CoV-2 (COVID19) corona viruses emerged from bats. Crimean Congo haemorrhagic fever (CCHF) and Ebola are currently the most important threats. After the first report of CCHF from Gujarat in 2011, it has been reported sporadically from Gujarat, Rajasthan and Uttar Pradesh. With current threats of CCHF, Ebola, Nipah, and other zoonotic viruses, ICAR-NIHSAD has an important role to play in conducting surveillance of these diseases in animals or vector hosts of these viruses, including wildlife. Since many of these emerging diseases have originated from animals, it is expected that more will follow unless a close watch is kept at the interface of animal human interaction. Anthropogenic factors are also responsible for facilitating the species jump of pathogens from wild animals to humans. The impact of any such pandemic, such as the COVID-19 in recent time, is tremendous and such episodes can perhaps be avoided through the One Health approach that the scientific world is now advocating. The animal health institutes of ICAR contributed to the fight against COVID-19 pandemic and are continuing it through the One Health approach to combat zoonotic disease threats to public health.
Conclusion
ICAR’s path-breaking innovative and cutting-edge technologies have helped the livestock and poultry industry to flourish and contribute extensively to raising the income of livestock owners. Animal Health Institutes of ICAR have developed more than 25 vaccines and about 80 diagnostics including nucleic acid-based tests, which have played a significant role in the control of economically important diseases and eradication of some of them from the country. In the current era of intellectual property rights, the adoption of new technologies from other sources requires various protocols. Hence, it is very important to develop indigenous technologies as has been emphasized by the Government of India through slogans ‘Vocal for Local’ and ‘Atmanirbhar Bharat’. Besides prevention and control of endemic diseases, the emergence of new diseases requires a robust disease forecasting mechanism, preparedness, and rapid emergency response to prevent the incursion of new transboundary diseases/ exotic diseases. Therefore, advanced preparedness to handle any such eventuality through prior risk assessment and rapid emergency response becomes very important to combat such incursions. Recent experiences with COVID-19 pandemic emphasize the need for robust epidemiological data and availability of indigenous and cost- effective diagnostics/ vaccine platforms for dealing with such outbreaks and pandemics. There is an urgent need for greater regional collaboration and cooperation with organizations like BIMSTEC, ASEAN, SAARC, etc., for sharing of information and technical know- how including microbial resources for monitoring and control of animal diseases.
By:Dr S.K. Malhotra, Project Director, Directorate of Knowledge Management in Agriculture, Indian Council of Agricultural Research, Krishi Anusandhan Bhavan-I, Pusa, New Delhi 110 012 and designed & printed at M/s Dolphin Printo Graphics, 1E/18, Fourth Floor, Jhandewalan Extension, New Delhi 110 055.
References
Ahuja V, Rajasekhar M and Raju R (2008) Animal Health for Poverty Alleviation: A Review of Key Issues for India. (https://www.dairyknowledge.in/sites/default/ files/ ahuja_et_al.pdf).
Alexander DJ, Aldous EW and Fuller CM (2012) The long view: a selective review of 40 years of Newcastle disease research. Avian Pathol 41(4):329-335. doi: 10.1080/03079457.2012.697991.
Annual Report (2016-2017 and 2017-18) National Research Centre on Equine, Hisar. Balamurugan V, Varghese B, Sowjanya Kumari S, Vinod Kumar K, Muthuchelvan D, Nagalingam
M, Hemadri D, Roy P and Shome BR (2021a. Avidin-Biotin recombinant nucleoprotein
competitive ELISA for the detection of peste des petits ruminants virus antibodies in sheep and goats. J Virol Methods 295:114213. doi: 10.1016/j.jviromet.2021.114213.
Balamurugan V, Vinod Kumar K, Dheeraj R, Kurli R, Suresh KP, Govindaraj G, Shome BR and Roy P (2021b) Temporal and Spatial Epidemiological Analysis of Peste Des Petits Ruminants Outbreaks from the Past 25 Years in Sheep and Goats and Its Control in India. Viruses 13(3), doi: 10.3390/v13030480.
Bardhan D (2007) India’s trade performance in livestock and livestock products. Indian J Agric Econ 62: 411-425.
Bardhan D, Kumar S, Anandsekaran G, Chaudhury JK, Meraj M, Singh RK, Verma MR, Kumar D, Kumar PTN, Ahmed Lone S, Mishra V, Mohanty BS, Korade N and De UK (2017) The economic impact of peste des petits ruminants in India. Rev Sci Tech 36(1):245-263. doi: 10.20506/rst.36.1.2626.
Bardhan D, Kumar S, Sekaran GA, Meraj M, Chilambarasan M, Singh R, Singh G, Pal R, Singh Y and Verma M (2020a) Economic losses due to hemorrhagic septicaemia in India. Indian J Anim Sci 90 (3):18-23.
Bardhan D, Kumar S, Verma MR and Bangar Y (2020b) Economic losses due to brucellosis in India. Indian J Comparative Microbiol Immunol Infectious Diseases 41:19. doi: 10.5958/0974- 0147.2020.00002.1
Bharadwaj M, Mandal BC and Rahal A (2020) Importance of Livestock in Indian Economy. Pashudhan Praharee (https://www.pashudhanpraharee.com/importance-of-livestock-in- indian-economy/).
DAHD (2015) Status of avian influenza in India, Department of Animal Husbandry Dairying and Fisheries, Govt. of India. (http://dahd.nic.in/dahd/WriteRead Data/Status%20of%20Avi an%20Influenza%20in%20India.pdf).
Dhingra MS, Dissanayake R, Negi AB, Oberoi M, Castellan D, Thrusfield M, Linard C and Gilbert M (2014) Spatio-temporal epidemiology of highly pathogenic avian influenza (subtype H5N1) in poultry in eastern India. Spat Spatiotemporal Epidemiol 11:45-57. doi: 10.1016/j. sste.2014.06.003.
Elschner MC, Laroucau K, H. Singha, B. N. Tripathi, M. Saqib, I. Gardner, S. Saini, S. Kumar, H. El- Adawy, F. Melzer, I. Khan, P. Malik, C. Sauter-Louis, and H. Neubauer. (2019). Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme- linked immunosorbent assays for serodiagnosis of glanders. PLoS One 14(4):0214963. doi: 10.1371/journal.pone.0214963.
Gargadennec L and Lalanne A (1942) La peste des petits ruminants. Bull des Serv Zootechniques et des Epizooties de l’Afrique Occidentale Francaise 5:16–21.
Garner M, Sawarkar SD, Brett EK, Edwards JR, Kulkarni VB, Boyle D and Singh SN (2000) The Extent and Impact of Sheep Pox and Goat Pox in the State of Maharashtra, India. Tropical animal health and production 32:205-223. doi: 10.1023/A:1005263601964.
Gemechu MY and Gill JPS (2011) Seroepidemiological survey of human brucellosis in and around Ludhiana, India. Emerging Health Threats Journal 4:7361, DOI: 10.3402/ehtj.v4i0.7361.
Govindaraj G, Vinayagamurthy B and Rahman H (2016) Estimation of Economic Loss of PPR in Sheep and Goats in India: An Annual Incidence Based Analysis. British J Virology 3:77-85. doi: 10.17582/journal.bjv/2016.3.3s.77.85.
Govindaraj GN, G Roy, BS Mohanty, V Balamurugan, AK Pandey, V Sharma, A Patel, M Mehra, SK Pandey and P Roy (2019) Evaluation of effectiveness of Mass Vaccination Campaign against Peste des petits ruminants in Chhattisgarh state, India. Transbound Emerg Dis 66(3):1349-1359. doi: 10.1111/tbed.13163
Iyer G and Hashmi Z (1945) Studies on Newcastle (Ranikhet) disease virus strain differences in amenability to attenuations. Indian J Vet Sci 15:155-157.
James A and Ellis PR (1978) Benefit-Cost Analysis in Foot and Mouth Disease Control Programmes.
British Veterinary J 134:47-52.
Kumar R, Kumar S, Khurana SK and Yadav SC (2013a) Development of an antibody-ELISA for seroprevalence of Trypanosoma evansi in equids of North and North-western regions of India. Vet Parasitol 196(3-4):251-257. doi: 10.1016/j.vetpar.2013.04.018.
Kumar S (1976) African Horse Sickness, ICAR Tech Bull 15:34.
Kumar S, Kumar R, Gupta AK, Yadav SC, Goyal SK, Khurana SK and Singh RK (2013b) Development of EMA-2 recombinant antigen-based enzyme-linked immunosorbent assay for seroprevalence studies of Theileria equi infection in Indian equine population. Vet Parasitol 198(1-2):10-17. doi: 10.1016/j.vetpar.2013.08.030
Kumar S, Rakha NK, Goyal L, Goel P, Kumar R, Kumar A and Kumar S (2015) Diagnostic application of recombinant equine merozoite surface antigen-1 in elisa for detection of Theileria equi specific antibodies. Jpn J Vet Res 63(3):129-137.
Newman SH, NJ Hill, KA Spragens, D Janies, IO Voronkin, DJ Prosser, B Yan, F Lei, N Batbayar, T Natsagdorj, CM Bishop, PJ Butler, M Wikelski, S Balachandran, T Mundkur, DC Douglas and JY Takekawa (2012) Eco-virological approach for assessing the role of wild birds in the spread of avian influenza H5N1 along the Central Asian Flyway. PLoS One 7(2):e30636. doi: 10.1371/journal.pone.0030636.
Otte J, Hinrichs J, Rushton J, Roland-Holst D and Zilberman D (2010) Impacts of avian influenza virus on animal production in developing countries. Cab Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources 3doi: 10.1079/PAVSNNR20083080.
Plowright W and Ferris R (1962) Studies with rinderpest virus in tissue culture: The use of attenuated culture virus as a vaccine for cattle. Res Veterinary Sci 3(2):172-182.
Rich K, Roland-Holst D and Otte J (2013) An assessment of the socio-economic impacts of global Rinderpest eradication-Methodological issues and applications to Rinderpest control programmes in Chad and India. Food Policy 44 doi: 10.1016/j.foodpol.2013.09.018.
Singh B, Prasad S, Sinha D and Verma MR (2013) Estimation of economic losses due to foot and mouth disease in India. Indian J Anim Sci 83(9):964-970.
Singh BK, Virmani N and Gulati B (2009a) Assessment of protective immune response of inactivated equine herpesvirus-1 vaccine in pregnant BALB/C mice. Indian J Animal Sci 79:345-349.
Singh RK, Balamurugan V, Bhanuprakash V, Sen A, Saravanan P and Yadav MP (2009b) Possible control and eradication of peste des petits ruminants from India: technical aspects. Vet Ital 45(3):449-462.
Singh RP and Bandyopadhyay SK (2015) Peste des petits ruminants vaccine and vaccination in India: sharing experience with disease endemic countries. Virusdisease 26(4):215-224. doi: 10.1007/s13337-015-0281-9.
Singh RP, BP Sreenivasa, P Dhar, RN Roy and SK Bandyopadhyay (2000) Development and evaluation of a monoclonal antibody based competitive enzyme-linked immunosorbent assay for the detection of rinderpest virus antibodies. Rev Sci Tech 19(3):754-763. doi: 10.20506/rst.19.3.1243
Singha H, K Shanmugasundaram, BN Tripathi, S Saini, SK Khurana, A Kanani, N Shah, A Mital, P Kanwar, L Bhatt, V Limaye, V Khasa, R Arora, S Gupta, S Sangha, H Sharma, SK Agarwal, J Tapase, S Parnam, P Dubey, SK Baalasundaram, BN Mandal, N Virmani, BR Gulati and P Malik (2020) Serological surveillance and clinical investigation of glanders among indigenous equines in India from 2015 to 2018. Transbound Emerg Dis 67(3):1336-1348. doi: 10.1111/tbed.13475.
Singha H, SK Goyal, P Malik, SK Khurana and RK Singh (2013) Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus. Indian J Virol 24(3):349-356. doi: 10.1007/s13337-013-0149-9.
Sreenivasa BP, P Dhar, RP Singh and SK Bandyopadhyay (2000) Evaluation of an indigenously developed homologous live attenuated cell culture vaccine against Peste-des-petits- ruminants infection of small ruminants. Proceedings of XX Annual Conference of Indian Association of Veterinary Microbiologists, Immunologists and Specialists in Infectious Diseases (IAVMI), Pantnagar, Uttaranchal, India, p 84.
Uppal PK and Yadav MP (1989) Occurrence of equine infectious anaemia in India. Vet Rec 124(19):514-515. doi: 10.1136/vr.124.19.514.
Uppal PK, Yadav MP and Oberoi MS (1989) Isolation of A/Equi-2 virus during 1987 equine influenza epidemic in India. Equine Vet J 21(5):364-366. doi: 10.1111/j.2042-3306.1989. tb02690.x
Vegad JL (2014) Drift variants of low pathogenic avian influenza virus: Observations from India.
World’s Poultry Science Jou 70:767-774. doi: 10.1017/S004393391400083X.
Virmani N, Bera BC, Singh BK, Shanmugasundaram K, Gulati BR, Barua S, Vaid RK, Gupta AK and Singh RK (2010) Equine influenza outbreak in India (2008-09): virus isolation, sero- epidemiology and phylogenetic analysis of HA gene. Vet Microbiol 143(2-4):224-237. doi: 10.1016/j.vetmic.2009.12.007.
Yadav M, Pandey A, Negi B, Sharma B and Shankar H (1986) Studies on inactivated goat pox vaccine. Indian J Virology 2(2):207-221.
Yadav M, Uppal P and Rao J (2016) Animal sciences. Singh RB (Ed) 100 Years of Agricultural Sciences in India. National Academy of Agricultural Sciences, New Delhi, p 158-258.
Yadav MP (2011) Laboratory Contributions for Rinderpest Eradication in India. Report submitted to FAO in Dec, 2011 under the Global Rinderpest Eradication Programme (GREP) (GCP/ GLO/302/EC), p 58.
