» Centre for Animal Disease Research and Diagnosis

Major Achievements

CADRAD was established in the year 1986 following the recommendations of QRT as an apex centre in the country for animal disease diagnosis. Various laboratories situated in different disciplines viz. Virology, Bacteriology, Parasitology, Toxicology, Pathology, etc. were combined together under the umbrella of CADRAD. Since its inception CADRAD is working excellently in the area of animal disease diagnosis and a total of 47,918 samples were analyzed in laboratories besides attending and investigating 112 outbreaks. Scientists, technical personnel’s and supporting staff work together as a team and analyze the samples taking full case as per the standard norms. This centre had also got ISO 9001: 2000 Certification for its Quality Management System and followed the standards.

Activity Milestone

1986	CADRAD established in IVRI, Izatnagar
1999	All India Conference of Disease Investigation Officers
2000	TOMBIT Workshop on ‘Biotechnology and Animal Health’
2000	ICAR Summer School on ‘Diagnosis of Emerging Diseases of Livestock’
2001	CDDL Scheme initiated by DADF, Govt. of India
2004	Epidemiology Division merged as a Section of CADRAD
2004	CADRAD was allotted separate present building
2005	Brain Storming Session organized on ‘TB, JD and STDs in Bovines’
2005	ISO 9001: 2000 Certification of CADRAD
2008	IAVP Conference was organized
2010	BSL-3 Laboratory sanctioned by DAHD&F, Ministry of Agriculture

Virology Laboratory

Facilities Available

• IBR- SNT, ELISA, PCR
• PPR- SNT, ELISA, PCR
• Rabies- FAT, RT-PCR, MIT
• Bluetongue- ELISA, RT-PCR, Virus isolation
• Classical Swine Fever- ELISA, RT-PCR
• Porcine Circovirus-2- PCR
• Porcine parvovirus- PCR
• Canine parvovirus- PCR, nPCR, HA,HI
• Canine distemper- RT-PCR
• Canine adenovirus- PCR

The Virology Laboratory was started with the objective to diagnose the viral diseases of livestock. The laboratory is having excellent cell culture facilities and maintaining a number of cell lines namely Vero, BHK21 MDBK, MDCK and CRFK. Besides, chicken embryo fibroblast cells are also used to isolate the viruses. Embryonated chicken eggs are often used to isolate the viruses by inoculating through a number of routes viz., CAM, I/V, allantoic, etc. The cell lines are sub cultured at regular intervals and preserved in liquid nitrogen for in house use and supply to other laboratories. A variety of tests namely AGPT for swine fever and bluetongue, competitive ELISA for IBR and bluetongue, indirect ELISA for IBR are also available. Virus neutralization test is used for screening of the serum samples for the presence of antibodies against IBR and bluetongue. PCR, a latest and highly sensitive test is available to detect the presence of viral DNA/RNA in the clinical samples such as (semen/ nasal swab / preputial washing) for IBR, classical swine fever, porcine parvovirus and porcine circovirus, canine adenovirus, canine distemper virus, blood samples for bluetongue and stool samples for CPV. Immunofluorescence tests are also available for the diagnosis of a number of canine viral diseases viz., canine parvo, canine distemper and canine coronavirus. Testing of potency of inactivated IBR vaccines in calves was done with limited success and an All India Network Programme on Bluetongue is being operated. Different inactivated vaccines using different adjuvants and inactivants are prepared and assessed in the sheep and are under laboratory trials.

During 1986-2006, a total of 15,589 samples including 7313 for IBR, 7476 for bluetongue, 221 for swine fever, 173 for rabies, 11 for cowpox, 57 for buffalo pox, 229 for PPR, 35 for sheep pox, 15 for goat pox, 15 for canine parvovirus and 4 for canine distemper were received for analysis and investigation.

BACTERIOLOGY LABORATORY

Facilities Available

• Tuberculosis- SID
• Johne’s Disease- SID, AGID, ELISA, PCR
• Brucellosis- RBPT, STAT, ELISA, PCR,Culture
• Leptospirosis- ELISA
• Campylobacteriosis- PCR, Culture
• Cultural Examination
• Antibacterial Sensitivity

As per the mandate and technical programme Bacteriology lab has meticulously investigated many disease outbreaks in animals. The lab has isolated several bacterial agents from clinical and morbid samples received / collected from animals all over the country. The animals of various organized Government and non-Government farms were tested for bacterial diseases. Moreover, this lab is also receiving samples from Zoological Parks and sanctuary, private organizations and different divisions and providing precise and timely diagnosis. Considering the importance of bacterial diseases among livestock, Government has identified four bacterial diseases, out of seven sexually transmissible diseases and policy framed is that the breedable animals should be free from STDs. Keeping in view this, the lab developed has facilities for detection of STDs agents utilizing molecular tools-like PCR and ELISA. Besides, lab has been doing culture, biological and antibiotic drug sensitivity assays routinely. The laboratory is also engaged in rapid diagnosis of Tuberculosis, Johne’s disease, Brucellosis, Campylobacteriosis, etc. A total of 20,790 samples were analyzed during 1986-2006, in the Bacteriology Laboratory. A total of 826 samples (preputial washings, semen, aborted foetuses and vaginal swabs) were screened for isolation of Campylobacter spp. of which 169 (20.5%) samples yielded Campylobacter jejuni (68), C. coli (27) and C. foetus (74). C. foetus is known pathogen of male and female reproductive system.

Presently, lab has been analyzing more than 5000 samples (clinical, morbid, serum, semen etc.) of different origins per year. Over the years lab has contributed precious infectious agents/ strains for future research and vaccine development. Different isolates of outbreaks are also maintained at this lab for further references and epidemiological studies. It has also suggested control measures at the face of outbreaks and prevented losses in livestock sector. The laboratory has been working for capacity building in the form of training and practical demonstrations to laboratory diagnosticians, microbiologists and field veterinarians of different states.

PATHOLOGY LABORATORY

Facilities Available

• Hematology
• Blood Biochemistry
• Urine, CSF, Synovial Fluid Examination
• Necropsy
• Histopathology
• Molecular Pathology
• Immunohistochemistry

The pathology laboratory is actively functional in CADRAD since its inception. During 1986-2006, a total of 10,945 samples were analyzed or cases investigated in pathology laboratory, consisting 7600 morbid samples, 2202 lab. animals, 390 aborted fetuses, 217 animals for postmortem diagnosis, 186 brain for rabies diagnosis and 350 brain for BSE diagnosis.

Disease diagnosis on morbid samples: A total of 7600 morbid samples consisting of 984 cattle, 1096 buffaloes, 1620 sheep, 555 goats, 396 pigs, 2143 dogs, 409 horses, and 397 wild animals were received for pathomorphological diagnosis. The samples were originated from all states of the country.

Disease diagnosis of Laboratory Animals: During the period of 20 years, a total of 2202 laboratory animals were received for postmortem examination and disease diagnosis from different Division of IVRI, Izatnagar and Mukteswar including Laboratory Animal Research Section. The laboratory animals mainly comprised of Guinea pigs, rabbits, rats and mice.

Aborted fetuses: During the period under report, a total of 390 aborted/ still born foetuses of cattle (356), buffaloes (12), sheep (12), goats (7) and pigs (3) were received for postmortem examination and laboratory diagnosis. All the foetuses were originated from Livestock Production and Management Division of this Institute. The important disease conditions diagnosed were Chlamydiosis, peritonitis, meningitis and still birth in sheep; still birth in goat; pnemoenteritis, enteritis, asphyxia, ascitis, head oedema, Staphylococcosis (S. albus and S. citrus), brucellosis (B. abortus and B. meletensis), Colibacillosis (E. coli), Proteus spp. and still birth in cattle; Traumatic injury in buffalo; and still birth in pigs.

PM diagnosis of animals including wildlife: A total of 217 animals including 25 cattle, 10 buffaloes, 25 sheep, 27 goats, 2 horses, 29 pigs, 85 dogs and 14 wild animals were received for postmortem and disease investigation.

Rabies diagnosis: A total of 186 rabies suspected samples were received for diagnosis. The samples were originated from UP, UK, MP, Maharashtra, AP, Delhi, Gujarat, Rajasthan and Nepal. The rabies was diagnosed in 9 cattle (4.8%), 6 buffalo (3.2%), 4 horses (2.15%), 53 dog (28.5%), 5 wild animals (2.69%), 1 camel and 1 human.

Detection by Seller’s and by FITC

BSE surveillance: BSE surveillance in the targeted cattle population (Native-born, crossbred and high risk cattle) was undertaken through Passive surveillance in which trainees, students were taught about BSE / scrapie and trained for brain collection, preservation and processing. During field visits also (outbreak investigation work, NATP, AHIS-DMS) efforts were made to educate the field veterinarians and farmers about BSE and to send brain samples from cattle dying of nervous signs. The active surveillance was carried out by collecting brain samples from cattle in postmortem room of the institute. Due to ban on cow slaughter in the UP state, brain samples from buffaloes, sheep and goats were studied. A total of 350 brain samples were collected/received in 10% formalin from crossbred cattle (10 PM cases), buffaloes (315 cases from local slaughter house), sheep (10 cases; 6 cases belonged to the institute and 4 of Rambouillet sheep of Pashulok Sheep Farm) and 5 goat. None of these animals showed BSE / scrapie compatible neurological signs (disturbances in behavior, sensitivity and locomotion). Two hundred seventy histological sections involving different neuro-anatomical sites, particularly obex, caudal cerebellar peduncle, rostral colliculi and other (spinal cord, pons, thalamus, pyriform lobe including hippocampus, cerebellar hemisphere, and cerebellum, etc.), were screened histopathologically for BSE / scrapie and other neurological disorders. Brain tissues did not show any appreciable microscopic lesions. A few showed vascular congestion, focal haemorrhage, perivascular oedema, at places focal glial cell reaction, neuronal degeneration and few vacuoles in white matter in some cases.

PARASITOLOGY LABORATORY

Facilities Available

• Faecal Examination
• Blood Test
• Parasite Identification
• Trichomonosis
• Skin Scraping Examination

Clinical Parasitology laboratory of Parasitology Division was transferred along with staff in CADRAD in, 1986 and was designated as Parasitology Laboratory. Various parasitic diseases were reported including Theileriosis, Babesiosis, Trypanosomosis; Immature Paramphistomosis, Schistosomosis, Fasciolosis; Roundworm infection, Lungworms Haemonchosis, Hookworm and Mange, tick infestation etc. Investigation revealed that in small ruminants Haemonchosis and large ruminants Fasciolosis and Trypanosomosis are major problem. From wildlife samples, Fasciola jaksoni was commonly prevalent in elephants while in tigers and leopards hookworm infection was encountered. A total of 8974 samples including 511 preputial washing, 6474 faecal, 1790 blood smear, 136 skin scrapings and 63 specimens were analyzed / parasites identified during 1986-2006.

TOXICOLOGY LABORATORY

Facilities Available

• Aflatoxin
• Ochratoxin
• Pesticide Toxicity
• HCN Poisoning
• Nitrate/Nitrite Toxicity
• Strychnine Poisoning

Considering the importance of toxicological investigations in disease diagnosis, Toxicology laboratory from Division of Pharmacology and Toxicology, IVRI, was merged in the Centre for Animal Disease Research and Diagnosis at time of its establishment in 1986. Field veterinarians report mortalities and morbidity in animals due to various factors other than specific diseases, which go undiagnosed. Toxicities / poisoning are often associated with such mortalities and morbidity in animals. Due to indiscriminate use of agrochemicals, increasing industrial/ environmental pollution, poisonous plants, malicious and accidental poisoning as well as poor managemental conditions farm and pet animals suffer from poisoning/ toxicity. These cause enormous economical losses to country’s animal industry and exchequer, which can be prevented by proper toxicological investigations based on the history, clinical symptoms, treatment attempted, the post-mortem lesions and toxicological tests/ animal experimentation data confirming the incriminating toxin using the knowledge and experience of toxicological problem. The timely diagnosis of toxicoses in animals based on the laboratory analysis is immensely useful in adapting ameliorative measures by specific antidotes so that animals could be saved. By adapting preventive measures, further toxicoses in animals can be prevented. A total of 3,574 samples from all parts of the country were received for analysis and results were conveyed durin1986-2006.

EPIDEMIOLOGY SECTION

• Diagnosis and epidemiology of Brucellosis
• Diagnosis and epidemiology of Bordetellosis
• Diagnosis and epidemiology of Glanders
• Diagnosis of tuberculosis
• Analysis of Data
• Sero-Epidemiology
• Epidemiology of emergence of drug resistance

EPIDEMIOLOGY SECTION

EPIDEMIOLOGY SECTION

EPIDEMIOLOGY SECTION

The Epidemiology Division was merged in CADRAD in the year 2004 as a Section. Facilities are developed for rapid diagnosis and understanding epidemiology of Glanders, Bordetellosis and Tuberculosis using conventional and PCR based methods. PCR based diagnosis for confirmation of genus and species of Burkholderia mallei has been standardized. Similarly, methods for isolation and characterization of Bordetella bronchiseptica from pigs and dogs have been standardized. Genus and species specific PCR for Bordetella bronchiseptica has already been developed. A PCR based differential diagnosis is available for Mycobacterium tuberculosis and M. bovis infection.

To acquire epidemiological information about brucellosis in small ruminant in Uttar Pradesh research is initiated. Under this project stratified random sampling will be done and from each district five villages will be selected. From each village, 25 sera samples from sheep and goats along with their information in prescribed proforma will be collected. During 2012-13, study will be carried out in Eastern UP, followed by in 2013-14 and 2014-15 in Central and Western UP, respectively. The study will also include the different small ruminant farms of the state. The information generated at the completion of the project will be shared with State Animal Husbandry Department along with suitable recommendations for prevention and control of this malady.

Besides, a repository of standard strains used in day to day work as reference has been created in the Section. Continuous work on antimicrobial sensitivity testing of clinical and outbreak strains lead to creation of a database which may be used in future for making general recommendations for the antibiotic prescriptions by the veterinarians. The recent trends revealed that the following recommendations may be useful for the veterinary practitioners.

Disease/ Pathogen	Sensitive (>90%), May be used	Resistant (>50%), Not to be used
Brucellosis	Tetracyclines, chloramphenicol, gentamicin	Fluoroquinolones (enreoflox), azithromycin, aztreonam, doxycycline, ampicillin, amoxycillin, amoxyclav, ampicillin+sulbactam
Edwardsiellosis	Azithromycin, aztreonam, ceftriaxone, fluoroquinolones, chloramphenicol	Tetracyclines, gentamicin, nitrofurantoin, ampicillin, amoxycillin, amoxyclav, ampicillin+sulbactam
Escherichia coli	Nitrofurantoin, gentamicin, ceftriaxone+tazatobactam, chloramphenicol, cefoperazone+sulbactam	Tetracyclines, gentamicin, nitrofurantoin, ampicillin, amoxycillin, amoxyclav, ampicillin+sulbactam
Klebsiella pneumoniae	Fluoroquinolones, gentamicin, imipenem, meropenem, cefoperazone+sulbactam, ceftriaxone+tazatobactam, aztreonam	Azithromycin, tetracycline, chloramphenicol, co-trimoxazole, nitrofurantoin, ampicillin, amoxycillin, amoxyclav, ampicillin+sulbactam
Moraxella bovis	Strepto-penicillin, and most of the common antibiotics	Aztreonam, ceftazidime, cotrimoxazole
Pasturellosis	Tetracycline, fluoroquinolones, amoxycillin, gentamicin, ceftriaxone	Chloramphenicol, co-trimoxazole, ampicillin, amoxycillin, amoxyclav, ampicillin+sulbactam
Salmonellosis	Chloramphenicol, gentamicin, ceftriaxone+tazatobactam, cotrimoxazole, imipenem	Tetracycline, nitrofurnatoin, cefoperazone+sulbactam, ampicillin, amoxycillin, ampicillin+sulbactam
Staphylococcal infections	Tetracycline, nitrofurantoin, chloramphenicol, amoxyclav	Ampiclox, aztreonam, ceftazidime+clavulanic acid, amoxy+sulbactam, vancomycin,
Streptococcal infections	Clindamycin, gentamicin, amoxyclav, vancomycin,	Streptopenicillin, ampicillin, fluoroquinolones, nitrufuron
Pseudomonas infections	No, use only after testing	Ampicillin, Streptopenicillin tetracycline, amoxy+sulbactam

CADRAD, being the National Referral Laboratory for Animal Disease Diagnosis is providing diagnostic services to Department of Animal Husbandry in all the states and union territories of India