(RESEARCH & TRAINING)

An IS/ISO 9001:2008 Organization

3377,, BBEELLGGAACCHHIIAA RROOAADD,, KKOOLLKKAATTAA--770000003377

2015 - 2016

NERDDL

NRDDL

CDDL

WRDDL

SRDDL

ERDDL

2

LOCATION OF RDDLs IN INDIA

3

STATES UNDER JURISDICTION OF REGIONAL DISEASE DIAGNOSTIC LABORATORY (ER)

With the establishment of Regional Disease Diagnostic Laboratory (ER), Kolkata diagnostic facilities have been extended to eastern & north-eastern states viz. Bihar, Orissa, Jharkhand, Sikkim and Andaman & Nicobar Islands.

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PREFACE

Since its inception, Regional Disease Diagnostic Laboratory (ER) has been efficiently

performing its role by concentrating on the specified mandate in areas of disease

surveillance, diagnosis, documentation, imparting training, maintenance of data bank on the

epidemiology of economically important diseases with the ultimate target of up-gradation of

livestock health.

All efforts were delivered for application of the latest diagnostic techniques for

diagnosis of different livestock and avian diseases. The Avian Influenza Laboratory has been

functioning effectively by screening of routine sera samples by competitive ELISA for

detection of Influenza Type „A‟ viral antibody and Haemagglutination Inhibition test for

detection of Influenza virus Subtype specific antibody. Routine screening of swab / tissue

samples for Avian Influenza Virus in Embryonated Chicken Eggs (ECE) is being carried out

purposefully. The competence of the laboratory for handling any unusual mortality sample

suspected for HPAI (Highly Pathogenic Avian Influenza virus) has been proved at several

instances. Recently, standardization of PCR for detection of Bacterial pathogens like Listeria

sp., Anthrax bacilli from blood and dried meat & HS specific Bovine Pasteurella sp. from

blood, Mycobacterium tuberculosis from tissue specimen, Mycoplasma sp. from nasal

secretion, milk, synovial fluid, and Babesia sp., Theileria sp., Anaplasma sp. from whole

blood had been done successfully. PCR has been standardised satisfactorily for diagnosis

of viral diseases like PPR, Lumpy Skin Disease, Goat Pox, Classical Swine Fever, Duck

Viral Enteritis and New Castle Disease also. qPCR technique is being routinely employed for

screening of Bovine Frozen Semen for BoHV-I viral DNA. Standardization of the pesticides

like Cypermethrin, Deltamethrin, Endosulfon, Quinalphos, BHC, Malathion, Mycotoxin like

Aflatoxin-B1, B2, M1, G2 and Ochratoxin has been done in HPLC Laboratory. Analysis of

Pesticide and Mycotoxin in specific samples has been done successfully. BSE surveillance

work and histopathological examination for diagnosis of several other diseases are being

performed regularly. Regular monitoring of the microbiological parameters of Frozen Semen

Bull stations, Bull Mother Farms of different states is executed as per MSP laid down by the

DADF, Government of India. Standardisation of PCR technique for BoHV-I has been

achieved and multiplex-PCR for different species of Brucella has been accomplished

previously.

Further, RDDL (ER), Kolkata is also imparting training to the officials attached to

disease diagnostic work of the dependant states and U/T especially on screening of serum

samples for sero-surveillance work against Avian Influenza, „PCR Technique‟ and upgrading

the knowledge against new techniques of different disease diagnosis including „Sexually

Transmitted Diseases in Bovine‟.

Having gathered the experience of this and previous years, we along with the co-

operation of all will strive to continue performing all the designated works with excellence in

the future too.

Sd/- (DR. RUDRADEB MUKHERJEA)

Joint Director, RDDL (ER) Institute of Animal Health & Vety. Biologicals (R&T)

Belgachia, Kolkata

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CENTRAL SECTOR SCHEME OF

DEPARTMENT OF ANIMAL HUSBANDRY & DAIRYING

REGIONAL DISEASE DIAGNOSTIC LABORATORY

(EASTERN REGION)

ANNUAL REPORT

2015-2016

1. Project Title Central Sector Scheme Animal Disease Management and Regulatory Medicine, Department of Animal Husbandry and Dairying, Ministry of Agriculture. Govt. of India.

2. Sanction No. F. No. 53-83/97-LDT (LH)

3. Date of Commencement July, 1999

4. Report Period 2015 - 2016

5. Head of Regional Disease Joint Director Diagnostic Laboratory Regional Disease Diagnostic Laboratory Eastern Region

6. Location Institute of Animal Health & Veterinary Biologicals (R&T) 37, Belgachia Road, Kolkata- 700037 An IS/ISO 9001:2008 Organization

Ph. : 033-2532-8033/033-2546-7087/033-2556-5021

Telefax : 033-2532-8033

e-MAIL ID : erddl_kolkata@yahoo.co.in

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REGIONAL DISEASE DIAGNOSTIC LABORATORY EASTERN REGION

M A N D A T E 1. To serve as regional referral laboratory for economically important livestock

diseases (bacterial, viral, parasitic etc.) and study of effects of any drugs/pesticides

residue/toxicants in the livestock products by providing diagnostic services (both

primary and confirmatory) to the states of the region.

2. To maintain a data bank on the epidemiology of the economically important diseases

of the region and circulate to all the states on a periodic basis.

3. Be responsible to collect and transmit all isolates of viral, bacterial and parasitic

agents to the National Repository to be maintained at the National Reference

Laboratory.

4. To follow the approved standard reagents and techniques for generation of the

diagnostic results.

5. To train scientists/officers from State Disease Diagnostic Laboratories of the region

to update different technology for diagnosis of animal diseases and provide

consultancy and expert services to the states of the region.

6. To undertake special programme for disease surveillance/diagnosis as per the

direction of Govt. of India from time to time in view of epidemiological significance

and importance etc.

7. Any data generated by the RDDL could be published only after prior permission

obtained from Department of Animal Husbandry, Dairying and Fisheries, Government

of India.

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STAFF PATTERN OF

REGIONAL DISEASE DIAGNOSTIC LABORATORY

EASTERN REGION, KOLKATA

Sl. No. Name of the post

Sanctioned strength

Present incumbent

1. Joint Director, RDDL (ER) 01 Dr. Rudradeb Mukherjea

Ph. D. ( Veterinary Surgery) (Acting)

2.. Specialist (Diagnostic Pathology) 01 Dr. Debasis Chatterjee

M.V.Sc. (Veterinary Pathology) (Acting)

3. Specialist (Diagnostic Virology) 01 Dr. Shamindra Nath Sarkar

M.V.Sc. ( Veterinary Virology)

4. Specialist ( Sero Epidemiology )

01

Dr. Joyjit Mitra

M.V.Sc (Veterinary Microbiology)

5. Specialist (Epidemiology &

Informatics) 01

Dr. Bidhan Bandyopadhyay

M.V.Sc. ( Veterinary Epidemiology & PVM)

6. Specialist (Diagnostic

Parasitology)

01 Dr. Rabindra Nath Sikari

M.V.Sc. (Veterinary Parasitology)

7. Specialist (Diagnostic Bacteriology

& Mycology)

01 Dr. Anup Singha

M.V.Sc.( Veterinary Bacteriology)

8. Specialist (Diagnostic

Biochemistry & Toxicology)

01 Dr. Sumit Chowdury

Ph. D. ( Veterinary Biochemistry) (Acting)

9. Laboratory Technician 09 Vacant

10. Laboratory Attendant 11 02

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ANNUAL ACTION PLAN AND TECHNICAL PROGRAMME

FOR RDDL (ER), KOLKATA

2015-2016

1. To provide referral and consultancy services to the state diagnostic laboratories for

economically important Livestock and poultry diseases.

2. Development of epidemiological Data Bank including epidemiological maps of economically

important livestock diseases of the region.

3. Screening of Bulls, teaser animals and bovine semen against sexually transmitted diseases

including estimation of microbial load in Frozen Semen Straw in the region.

4. qPCR based diagnosis with special reference to detection BoHV-1 in bovine semen sample.

5. Multiplex PCR for diagnosis of Brucella sp.

6. Surveillance work of infectious causes of abortion in Livestock population in organized farms

and in the rural areas.

7. Surveillance work on BSE based on Histopathological studies and ELISA based diagnosis of

brain samples (in accordance with the laid down target of Govt. of India).

8. Screening of brain samples from Bovine Sp. exhibiting neurological disorders for Listeriosis.

9. Immune status studies on Ranikhet Disease in vaccinated organized and backyard poultry

population.

10. Application of molecular biological techniques for routine diagnosis of different livestock &

poultry diseases (other than Avian Influenza) with special reference to Capri pox , PPR,

Classical Swine Fever, Duck Viral Enteritis, Duck Viral Hepatitis, New Castle Disease,

Brucellosis, Listeriosis, Anthrax, HS, CCPP, Tuberculosis. etc.

11. Routine application of PCR techniques for diagnosis of haemoprotozoan infection in cattle like

Babesiosis, Anaplasmosis and Theileriosis etc.

12. Routine Sero-surveillance work and antigen detection in swab / tissue samples of HPAI.

13. Surveillance of Avian Influenza virus as per guideline of Govt. of India and framed surveillance

strategies adopted by the different states under jurisdiction of RDDL (ER), Kolkata.

14. Diagnosis of Rabies infection in post-mortem material by immuno-chromatographic technique.

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15. Standardization of ELISA technique for detection of Mycotoxin in animal and poultry feed.

16. Proximate Analysis of animal and poultry feed.

17. To impart analytical facilities of pesticides, antibiotics and drug residues in livestock and poultry

products by HPLC.

18. A) Study of nutritional status in infertility cases.

B) Clinical Haematology and Bio-chemistry to aid disease diagnosis.

19. Soil testing of Anthrax spore in the endemic area.

20. Testing of export materials as per OIE protocol.

21. Maintenance of Repository of infective agents isolated from the Eastern Region.

22. Organizing training programme for veterinary & para-veterinary work force.

23. Preparation of different Audio-visual presentation for Mass awareness of vets/para-vets

and common people against economically important livestock and poultry diseases.

24. Detection of microbial agents responsible for clinical and sub-clinical mastitis and its

antibiogram.

25. Monitoring of State Sponsored Scheme on disease investigation.

26. Monitoring of Centrally Sponsored Scheme of NPRE/AICRP on FMD.

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ACTIVITY REPORT OF RDDL (ER)

2015-2016

A. Maintenance of Data Bank on epidemiology on various Disease encountered in the Region (as per technical programme-2) :

DISEASE OUTBREAK REPORTS OF EASTERN REGION

Sl. No.

Disease West

Bengal Bihar Odisha

Andanam

& Nicobar Islands

Other States

under the jurisdiction

of RDDL(ER)

1. Anthrax 06 0 19 0

Report not yet

Received

2. Anaplasmosis 08 03 01 04

3. Avian Influenza 0 0 0 0

4. Avian Salmonellosis 02 08 0 17

5. Babesiosis 03 04 02 03

6. Black Quarter 22 02 18 0

7. CCPP 0 0 0 34

8. CRD 0 0 02 14

9. Duck Cholera 0 0 0 04

10. Duck Plague 10 0 0 0

11. Foot & Mouth Disease

23 04 20 0

12. Fowl Cholera 02 01 0 17

13. Fowl Pox 12 02 0 0

14. Goat Pox 13 03 0 0

15 H.S. 06 02 07 0

16 I.B.D. 04 02 01 26

17 PPR 126 01 05 0

18 Rabies 01 0 0 0

19 Ranikhet 42 07 01 29

20 Swine Fever 03 04 01 0

21 Theileriosis 04 0 02 05

22 Trypanosomiasis 02 0 02 04

23 Coccidiosis 05 03 01 28

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12

13

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B. To provide referral and consultancy services to the state diagnostic

laboratories for economically important livestock and poultry diseases

(as per technical programme-1) :

1. Provided diagnostic services to Frozen Semen Bank, Cuttack, Orissa against

Brucellosis, Campylobacteriosis, Trichomoniasis.

2. Imparted sero diagnostic support against Brucellosis, IBR of samples from CCBF,

Koraput, Orissa, CCBF, Chiplima, Orissa and Bihar Livestock Development Agency,

Patna.

3. Provided sero diagnostic support against Leptospira for Andaman and Nicobar Islands.

4. Provided diagnostic support in connection with PPR outbreak in Eastern Region.

5. Provided diagnostic support in connection with Anthrax outbreak among livestock and wild

life in Eastern Region particularly in the state of Odisha.

6. Provided diagnostic support in connection with CCPP for Andaman & Nicobar Islands.

7. Performed Sero monitoring under National Sero Surveillance Programme on FMD for the

state of Sikkim.

8. Performed examination of all the serum and swab / tissue samples for surveillance of

Avian Influenza received from different states under jurisdiction.

9. Imparted „Training on Avian Influenza Diagnosis‟ to the officers of different states against

proposal of Station In-charge, IVRI Eastern Regional Station, Kolkata, after getting

instruction from the competent authority.

10. Organized Residential Training on „Advanced Laboratory Technique for Screening of

Sexually Transmitted Diseases in Bovine‟ and „PCR Technique for Diagnosis of Common

Bacterial, Viral & Parasitic Diseases of Livestock‟ as per the mandate of RDDL for officers

of different states under jurisdiction of RDDL (ER).

11. Students of final year B.V.Sc & A.H. from WBUAFS, Kolkata attended to different section

of this Institute in batch wise for training on practical aspect of disease diagnosis.

12. Provided diagnostic services to Frozen Semen Bank, Cuttack, Orissa, CCBF, Koraput,

Orissa, CCBF, Chiplima, Orissa, Livestock Breeding & Dairy Farm, Cuttack, Odisha,

against Tuberculosis and Johnin.

13. Provided diagnostic support against Tuberculosis for the state of Bihar.

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C. Screening of bulls, teaser animals and bull semen against sexually transmitted

diseases including estimation of microbial load in frozen semen straw (as per technical programme-3, 4, 6) :

As per mandate of ERDDL different samples were received/collected from organized/unorganized farms in West Bengal and other States during this period. Different tests undertaken in the laboratory during the period of April, 2015 to March, 2016 and results are shown in different tables.

C-1. Serological examination reports of different sexually transmitted diseases :

Source

Brucellosis IBR

Remarks Total Sample

s P N

Total Samples

P N

Organized Farms of

West Bengal 1969 248 1721 1276 894 382

P= Positive N= Negative

Unorganized Farms of

West Bengal

573 01 572 30 05 25

OTHER STATES

Central Cattle Breeding Farm, Koraput,Odisha

167 76 91 - - -

Frozen Semen Bank, Cuttack,

Odisha 106 - 106 106 56 50

ADDL, Dollygung

A & N Islands 95 - 95 - - -

Bihar Livestock Dev. Agency ,

Patna, Bihar 24 - 24 24 - 24

National Dairy Dev. Board, Hyderabad

20 10 10 20 10 10

TOTAL 2954 335 2619 1456 965 491

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C-2. Serological examination on Bovine Viral Diarrhoea (BVD) for Antibody Detection :

Name of the Farm/ Place

Total No. Animals Tested

No. of Positive Animals

No. of Negative Animals

Organized Farms of West Bengal

229 75 154

Frozen Semen Bank, Cuttack, Odisha

106 76 30

Bihar Livestock Dev. Agency , Patna, Bihar

12 - 12

National Dairy Dev. Board, Hyderabad

20 12 8

TOTAL 367 163 204

C-3. Serological examination on Bovine Viral Diarrhoea (BVD) for Antigen Detection :

Name of the Farm/ Place

Total No. Animals Tested

No. of Positive Animals

No. of Negative Animals

Organized Farms of West Bengal

1146 6 1140

SDDL of other States Samples not received - -

TOTAL 1146 6 1140

C-4. Estimation of microbial load in Frozen Semen straw (as per technical programme-3) :

Source Total Sample tested Satisfactory Unsatisfactory

Frozen Semen Bull Stations/ Sperm Stations West Bengal

825

764

61

SDDL of other States Samples not received - -

TOTAL 825 764 61

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D. Surveillance work on infectious causes of abortion in livestock population

(as per technical programme-6) :

D-1. Cultural examination reports on different sexually transmitted diseases :

Source

Type of

samples

Total

samples

received

Campylobacteriosis Trichomoniasis

Remarks P N P N

Frozen Semen Bull Stations /

Sperm station West Bengal

Preputial washing

226 - 226 - 226 P= Positive N= Negative

Frozen Semen Bank,

Cuttack, Odisha 108 - 108 - 108

Bihar Livestock Dev. Agency,

Patna, Bihar 24 - 24 - 24

TOTAL 358 - 358 - 358

E. q-PCR based detection :

E-1 q-PCR based detection of BoHV - I in neat semen

(as per technical programme Sl. No. 4) :

As per direction laid down in the “OIE Terrestrial manual (2009) for standardization of Diagnostics and vaccines”, the recommended tests for International trade for IBR is Real time (quantitative) -PCR technique. As per proposed MSP of Govt. of India for application of q-PCR, a Resolution has been taken in mid-year of 2012-2013, which was incorporated in MSP. We have standardized the test in this current year 2014 - 2015. Results of q-PCR based detection of frozen semen straws in batches during the year 2015-2016 are stated below :

Source Total Sample tested

(in batches) Positive

Negative

Frozen Semen Bull Stations/ Sperm Stations West Bengal

4203 133 4070

SDDL of other States Samples not received - -

TOTAL 4203 133 4070

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F. Surveillance work on Tuberculosis :

During the year 2015-2016, the Instt. of Animal Health & Vety. Biologicals (R&T), Kolkata, as per RDDL mandate, tested 1758 nos. of animals for tuberculosis in bovine population and in wild life in different states under jurisdiction of RDDL(ER) using single interdermal test by tuberculin PPD. One animal was found reactor. In case of Johne‟s disease there was no reactor animal out of 1645 animals tested in different states under jurisdiction of RDDL(ER).

F-1. Tuberculin Sensitivity Test at Govt. Farms/ Private Farms/ organization :

Sl. No.

Name of the Farm/Institution No. of animal Tested

No. Of Reactor Animal

No. of Doubtful Animal

No. of Non - Reactor animal

WEST BENGAL

1 Sri Mayapur Goshala Trust, Mayapur, ISCON

83 1 4 78

2 Bull Mother Farm & Exotic Cattle Breeding Farm, Salboni, Mednipur (W)

139 - - 139

3 Frozen Semen Bull Station, Salboni, Mednipur (W) 107 - - 107

4 Bull Mother Farm, Haringhata 529 - 4 525

5 Sperm Station, Beldanga, Murshidabad 30 - - 30

6 Frozen Semen Bull Station, Haringhata 114 114

7 Animal Rescue &Transit Facility Centre( WARTFC), Saltlake, Kolkata

12 - - 12

8 State Quarentine Centre, Fulia, Nadia 14 14

9 Animal Rescue &Transit Facility Centre (WARTFC) , Saltlake, Kolkata

14 - - 14

10 FSBS, Salboni, Mednipur ( W ) 113 - - 113

11 Serm Station, Beldanga, Murshidabad 27 - - 27

ODISHA

12 Central Cattle Breeding Farm, Chiplima 4 - - 4

13 Central Cattle Breeding Farm, Sunabeda-2, Koraput 213 - - 213

14 Frozen Semen Bank, Khapuria , Cuttak 108 - - 108

15 Central Cattle Breeding Farm, Chiplima 251 - 4 247

TOTAL 1758 1 12 1745

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F-2. Johnin Sensitivity Test at State Farms/ Private Farms/ organization :

F-3. Total Test Performed :

Sl. No.

Name of the Test No. of Animal Tested

No. of Reactor animal

No. of Doubtful animal

No. of Non-reactor animal

1 Tuberculin Sensitivity Test 1758 1 12 1745

2 Johnin Sensitivity Test 1645 - 2 1643

TOTAL 3403 1 14 3388

G. Surveillance work on BSE by Histopathological studies and ELISA based diagnosis

of brain samples (as per technical programme-7) :

Active Surveillance :

A total of one hundred fifty (150) brain samples (mostly cattle) were collected or received in 10% formalin or ice pack at different slaughter houses from West Bengal, Bihar. Besides, sample from clinically suspect cases were collected /received from different places of West Bengal, Sikkim, Jharkhand, Andaman & Nicobar, Orissa, Bihar and Tripura exhibiting neurological sign before death.

Sl. No.

Name of the Farm/Institution No. of Animal Tested

No. of Reactor Animal

No. of Doubtful Animal

No. of Non Reactor Animals

WEST BENGAL

1 Bull Mother Farm & Exotic Cattle Breeding Farm, Salboni, Mednipur ( W )

139 - - 139

2 Frozen Semen Bull Station, Salboni, Mednipur (W) 107 - - 107

3 Bull Mother Farm, Haringhata 529 - 2 527

4 Sperm Station, Beldanga, Murshidabad 30 - - 30

5

Frozen Semen Bull Station, Salboni, Mednipur (W) 113 - - 113

6 Sperm Station, Beldanga, Murshidabad 27 - - 27

7 State Quarentine Centre, Fulia, Nadia 14 - - 14

8 Frozen Semen Bull Station, Haringhata 114 - - 114

ODISHA

9 Central Cattle Breeding Farm, Sunabeda-2, Koraput 213 - - 213

10 Frozen Semen Bank, Khapuria , Cuttak 108 - - 108

11 Central Cattle Breeding Farm, Chiplima 251 - - 251

TOTAL 1645 - 2 1643

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The coronal section from different anatomical sites particularly obex of medulla, caudal cerebellar peduncle, rostral colliculi and others were cut and fix immediately in 10% formalin to avoid autolytic changes. After fixation tissues were processed, cut and stained (H&E) and Seller‟s per standard procedure for microscopical examination. Ice pack tissues were screened to detect Listeria species and Rabies (negri bodies) by Seller‟s staining.

None of the brain sample screened had hallmark lesion suggestive to BSE. Though some of the samples taken from clinical suspect cases having few nervous sign showed mostly vascular or neurodegenerative changes.

All of the brain samples screened for rabies and Listeriasis were also found negative.

G-1. Target and achievement of receiving Bovine Brain Samples (clinical suspect cases) from different States of ERDDL :

Sl. No.

Name of the States

Target from clinical suspect case

No of samples received Shortfall of clinical suspect cases

Slaughter house Clinical suspect case

1. Bihar 17 10 - 17

2. Jharkand 09 - - 09

3 Orissa 16 - 23 -

4. Sikkim 05 - 05 -

5. Tripura 05 - 06 -

6 West Bengal 20 80 20 -

7 A & N Island 02 - 06 -

TOTAL 74 90 60 26

G-2. General Characteristic of Animals Presented at Different Slaughter house / Farm/ Village under ERDDL :

Species Bovine

Breed Indigenous, Sahiwal, Gir, Red Sindhi Jersy Cross and

Holstein-Friesian Cross, Murrah & Chilika

Age Above 2 yrs. to 10 yrs.

Sex Male & Female

Place of Origin Different parts of West Bengal, Bihar, Andaman

& Nicobar Island, Orissa, Sikkim & Tripura

Health Status 80% of animals were apparently healthy

and 20% animals health conditions are very poor.

External Appearance & Fitness For Work

Mostly old aged, weak, emaciated debilitated and unfit for work and milking purpose.

Health Status of Animals from field Condition

Apparently normal, but animals had neurological symptoms of different diseases

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G-3. Overall progress of diagnosis of different neurological diseases with special reference to BSE/ TSE diagnosis & surveillance done by ERDDL :

G-4. Histopathological Interpretation of Bovine Brains :

G-5. Surveillance Point values for samples collected from animals in the given Sub population & age Category in the Bovines of Eastern Region at ERDDL

Sl. No. Parameter Total Source / Result

1. Total no. of brains collected 150 Abattoirs & fields

2. Brains collected from abattoir 90 -

3. Brains collected from fields 60 -

4. Number of brains processed 150 -

5. Number of brain screened for histopathology 150 Negative to BSE hallmark lesion

6. Total number of brain send for

screening of Listeriosis 31 Negative to Listeriosis

7. Brain examined for Rabies by Seller’s staining 22 Negative to Rabies

Sl No.

Histopathological Observation No. of Brains

Percentage of lesion (%)

Remarks

1. Total No. of Brains collected 150 - Different Abattoirs & fields of West Bengal, Andaman & Nicobar Island, Bihar, Jharkand, Orissa ,Sikkim

2. No. of brains screened 150 - -

3. Chronic encephalopathy 08 5.33

Special emphasis should be given for

collection of brain samples from clinically

suspected cases of, Bihar, Jharkhand, Sikkim

as per GOI guideline, otherwise the target

will not be achieved in future

4. Vasculo encephalopathy 14 9.33

5. Rabiform encephalopathy 02 1.33

6. Bac./Viral/Toxic infective

encephalopathy 05 3.33

7. Low grade encephalopathy 32 21.33

8. Tumors/ cyst/ parasitic lesion - -

9. No significant lesion 89 59.33

Summary Table for BSE Surveillance

Surveillance Sub-population

Age Routine Slaughter Fallen Stock Casualty Slaughter Clinical Suspect Total

Sample Points Sample Points Sample Points Sample Points

>1&<2 yrs - - - - - - - 0

>2&<4 yrs 00 00 - - - - 05 1300

>4&<7 yrs 42 8.4 - - - - 50 37500

>7&<9 yrs 33 3.3 - - - - 04 880

>9 yrs 15 00 - - - - 01 45

TOTAL 90 11.7 - - - - 60 39725 39736.7

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G-6: Audit findings in feed mills producing feed for ruminants (inspections and sampling)

Type of plant Number

of plants

Number of plants in (A) inspected

Total number of

visual inspections

in (B)

Total number of

plants in (B) with

infarctions

Total number of inspected plants

in (B) with sampling

Total number of plants in (C) with positive test results

(A) (B) (C)

For ruminants only 13* 6 6 - 1 #

For multispecies 01 - - - - -

* State Run Feed Mill Plant In West Bengal 12 Nos and in Bihar 1 No. (used MBM) and report from other

states are not yet received.

# Feed samples were sent to SRDDL on 17/03/2016, but result yet to be received.

H. Immune status studies on Ranikhet Disease in vaccinated organized and backyard poultry population in the region (as per technical programme 9) :

To study immune status in organized poultry farms following scheduled vaccination against Ranikhet disease as well as sero survey work against Pullorum disease were conducted during the period. A total of 4018 no. of serum samples were received from backyard and organized farms of West Bengal, out of which 2063 (51.34%) samples exhibited protectivity against Ranikhet disease.

H-1. Results of serum antibody against Ranikhet & Pullorum diseases :

Source of sample

No. of Sera

sample collected

No. of sera sample tested

Test Performed

Result for Ranikhet Disease

Result of Serological test for S. pullorum

Protective

(>1:32)

Non-protective

(<1:16)

West Bengal

(Backyard and organized Farm)

4018

4018

HA, HI

2063 (51.34%)

1955

(48.66%)

132 samples were shown sero-positive (3.43%) for

detection of Salmonella Pullorum out of 3842 samples tested

Samples from other States Nil Nil Nil - - -

23

The absence of development of protective titer to the desired level among immunized birds for R.D. Immune Status Studies may be due to various reasons. The improper maintenance of cold chain, presence of immune suppressive factors (as IBD infection) or other consistent infection may play some role in it.

I. Routine sero-surveillance work and Antigen detection in swab / tissue samples of HPAI in the region (as per technical programme-12 and 13) :

Avian Influenza Laboratory was established to detect an area affected with Avian Influenza in quickest possible time to prevent further spread, also to carry out routine surveillance diagnostic work of Avian Influenza in different states under jurisdiction of RDDL(ER) with the intention to verify that an area is free from Avian Influenza infection or not.

Monthwise serum samples received from West Bengal during 2015-16

24

I-1. Work undertaken and Reports during the period :

i. Receiving, cataloguing, pooling and numbering of samples for surveillance of Avian

Influenza.

ii. Serological examination of samples by competitive ELISA for detection of Influenza virus

Type A antibody.

iii. Serological examination of sera samples positive for Influenza Type A viral antibody by Haemagglutination Inhibition (HI) Test for detection of Influenza virus Subtype H5/H7/NDV etc. antibody.

iv. Diagnosis of randomly selected swab / tissue surveillance samples and detection of viral antigens of Influenza virus Type A by Immuno-chromatographic Test (Rapid AIV Test kit).

v. Examination of swab / tissue surveillance samples for Avian Influenza Virus in Embryonated Chicken Eggs (ECE) and HA test.

vi. Detection of Avian Influenza viral nucleic acid by Real Time RT PCR test in suspected swab / tissue samples of unusual mortality among poultry birds.

vii. Conventional RT-PCR test for detection of Avian Influenza virus if necessary. viii. Preparation of logistics for sample collection like sample transport media, dispensing and

distribution of logistics for serum and swab collection on indent in regular basis.

Though a total of 61,107 numbers of samples were received in the Laboratory form different states for routine surveillance diagnosis during the period 2015-16. Sera Samples received for surveillance from different states under jurisdiction of ERDDL are 17,198 (fowl sera- 13,255, duck sera-3,943). Swab / tissue Samples i.e. cloacal swab, tracheal swab, faecal samples from migratory birds, environmental samples (drinking water, waste water, droppings, processing tables, processing knives etc) received for surveillance of Avian Influenza are 43,909.

Avian influenza surveillance programme has been done and will be continued as per national surveillance plan provided from DADF,Govt. of India. As per existing new national surveillance plan on Avian Influenza as framed by Government of India each and every state under RDDL(ER) will follow strictly the plan during sample sending. A special emphases was given on collection and testing of samples from the hot-spot areas viz the national international border areas, areas of previous occurance, water bodies and wild life sanctuaries and specially the Live Bird Markets (LBM). A total of 8576 samples were collected from LBM from different districts of which the number of sera samples were 2144 and swab / tissue (swab/tissue) samples were 6432. In addition to it, 8718 number of environmental samples and 845 faecal materials from migratory birds were tested in the laboratory. The different tests undertaken in the laboratory during the period of April, 2015 to March, 2016 for examination of the sera samples were Competitive ELISA and Haemagglutination Inhibition tests. Of these, AIV Ab ELISA detects Influenza virus (Type A) positive sera while Haemagglutination Inhibition test (using H5/H7/H9/NDV Subtype specific antigen) could detect Avian Influenza virus specific positive sera samples.

25

The recommended Haemagglutination Inhibition (HI) test (using H5/H7/H9/NDV Subtype specific antigen) was carried out for cross-checking by taking randomly a certain percentage of serum received from different states. A total of 19,342 numbers of serum samples has been screened by ELISA in the year 2015 -16.

Haemagglutination Inhibition Test was carried out for screening of the sera samples individually which was a cumbersome task. A certain percentage of samples which is very less compared to large numbers of serum samples received has been tested by Haemagglutination Inhibition with standard sera for cross checking as well as for calibration and validation of test protocols.

In respect of examination of routine swab / tissue samples, Embryonated Egg Inoculation technique has been done in routine basis and „Egg Inoculation test‟ followed by Slide agglutination test and if necessary, Haemagglutination Inhibition test using H5/H7/NDV specific antisera has been carried out. It was observed that the Chromatographic Immunoassay using Anigen® Rapid AIV Ag test kit has a fast and fairly acceptable detection limit. Therefore, initial screening of the swab / tissue samples by this kit can reduce handling of the large number of embryonated eggs. This may allow the laboratory worker to devote more time and energy for subsequent virological examination of the eggs after inoculation with only those samples that exhibited positivity in Chromatographic Immunoassay. We are expecting, the above SOP for handling routine surveillance swab / tissue samples. However, this will need enough number of Chromatographic Immunoassay kits (Rapid AIV Ag test kits) in the year 2016-17. But, disadvantage is - Chromatographic Immunoassay exhibits positive result irrespective of virus survivability.

For examination of swab / tissue samples of unusual mortality, „Real Time RTPCR test‟ can be carried out as a quick and confirmatory test for detection of viral nucleic acid of Avian Influenza virus in addition to „Egg Inoculation test‟ and „Chromatographic Immunoassay‟.This proves that the laboratory is ready to handle any unusual mortality samples suspected of HPAI. This proves that it was worth establishing the Pre-Fabricated BSL-III laboratory and its relevant instruments.

Still then, maintaining a BSL-III laboratory is an expensive endeavour. Though standardization of the required techniques for avian influenza diagnosis (involving several costly instruments, chemicals and biological) could be successfully accomplished, utilization of the valuable Pre-Fabricated BSL-III laboratory along with the costly instruments for the purpose of diagnosis of Avian Influenza alone does not stand commendable. Therefore, it is proposed to allow this laboratory for standardization of diagnostic methods for other animal pathogens by molecular techniques.

26

The activities performed in the laboratory during the period under reporting are tabulated here :

I-2. Details of fit samples received in Avian Influenza Laboratory :

I-3. Details of Live Bird Market samples of West Bengal tested in Avian Influenza Laboratory :

Particulars No.

Number of Districts covered 18

Number of Locations covered 124

Number of Serum collected and tested 2144

Number of Swab samples collected and tested 6432

Number of environmental samples collected and tested 8191

I-4. Details of Serum Samples tested by ELISA :

State

No. of Fowl

serum tested

No. of Fowl

positive samples

No. of Duck serum tested

No. of Duck

positive samples

Other serum tested

No.of other

positive samples

Total Serum tested

Total No. positive samples (influenza type A)

A &N Islands - - 75 - - - 75 -

Bihar 2814 - - - - - 2814 -

Sikkim 5 - - - - - 5 -

Odisha 1646 - - - - - 1646 -

Tripura 1230 - 276 - - - 1506 -

West Bengal 9704 - 3592 - - - 13296 -

TOTAL 15399 0 3943 0 0 0 19342 0

State

Serum Cloacal Swab Tracheal

Swab Environmental

sample

Migratory bird

Faecal sample

Live Bird Market (LBM)

TOTAL

Fowl Duck Fowl Duck F Fowl Duck - - Serum

Swab / tissue (swab/ tissue)

-

A & N Islands

- 75 109 - 109 - - - - - 293

Bihar 2814 - 3381 1096 203 510 8004

Jharkhand - - - - - - - - - - -

Odisha 1646 - 6281 1037 - 324 179 - - 9467

Tripura 1230 276 846 300 924 36 - - - - 3612

Sikkim 5 - 10 - - - - - - - 15

West Bengal 7560 3592 4978 2148 4474 41 8191 156 2144 6432 39716

TOTAL 13255 3943 15605 2448 7640 77 8718 845 2144 6432 61107

27

I-5. Details of Serum samples tested by Haemaglutination Inhibition test :

I-6. Details of Swab / tissue (swab/tissue) samples tested by Egg Inoculation

Technique in he Avian Influenza Laboratory :

I-7. Details of Swab / tissue samples / Harvested Allantoic Fluid tested by Chromatographic Immunoassay in Avian Influenza Laboratory :

State Total Swab /

tissue samples

tested

Total No. of

+ve samples

Bihar 68 -

Sikkim 10 -

Tripura 76 -

A&N Islands 12 -

Odisha 148 -

West Bengal 256 -

TOTAL 570 0

State Fowl

serum

No. of Fowl positive samples

Duck serum No. of Duck

positive samples Total No. of

Positive sample

A & N Islands 0 - 5 - -

Bihar 540 - - - -

Sikkim 5 - - - -

Odisha 395 - - - -

Tripura 290 - 60 - -

West Bengal 1540 - 495 - -

TOTAL 2770 0 560 0 0

State Fowl No.of +ve samples

Duck No.of +ve samples

Other No.of +ve samples

Total No. of Swab / tissue (swab/tissue)

samples

Total No. of +ve samples

A & N Islands

218 - - - - - 218 -

Bihar 5190 - - - - - 5190 -

Sikkim 10 - - - - - 10 -

Tripura 1770 - 336 - - - 2106 -

Odisha 7642 - - - 179 - 7821 -

West Bengal

24075 - 2189 - 156 - 26420 -

TOTAL 38905 0 2525 0 335 0 41765 0

28 J. Detection of Mycotoxin and Proximate Analysis of Animal and Poultry feed/feed

ingredients (as per technical programme-16) :

Mycotoxin and Proximate Analysis Lab. at IAH &VB (R&T) Kolkata is engaged in the analysis of various feed and feed ingredients for various parameters like Moisture, Ash, Fibre, Fat etc. as per requirement of the farmers/farms. Four feed samples from Bihar, were received for proximate analysis during the year 2015-2016 at ERDDL, Kolkata but remain untested due to non-functionality of required instruments.

K. To develop analytical facilities of pesticides, antibiotics and chemical residues in Livestock & poultry products (as per technical programme-17) :

As per mandate of ERDDL, pesticides, antibiotic and drug residues in livestock and poultry products are to be determined. For the purpose, a laboratory is established and the necessary equipments like HPLC, standards chemicals and reagents have already been procured earlier. Now the laboratory is functioning with the following activities.

K-1. Standardization of Pesticides :

Standardization of the pesticides like Cypermethrin, Deltamethrin, Endosulfon, Quinalphos, BHC, Malathion, Monocrotophos and Phosphomidon has been done.

K-2. Standardization of Mycotoxins :

Standardization of Mycotoxins like Aflatoxin-B1, B2, M1, G2 and Ochratoxin has been standarized .

K-3. Sample analysis :

Eighty eight (88) tissue samples for analysis of Pesticides and twenty five (25) feed samples for Mycotoxin analysis has been done during the period. Sample analysis report is as follows:

No. of Sample

Nature of Sample Purpose Analytical Results

25

Feed materials from Govt.

Poultry Farms &

Tissue materials from DI Lab.

Mycotoxin

Analysis

Detected Aflatoxin B1,

B2, G2, M1 and

Ochratoxin found below

Permissible Limit

88

Tissue sample from Bacterial

Disease Lab., DI Lab., from

different District of west Bengal

and from other States

Pesticide Residue

Analysis and to find out

non-infectious causes

of death.

Two samples found

positive for Malathion

& Endosulfon etc. and

others are negative.

29

L. Study of Nutritional Status in infertility cases (as per technical programme-18) : Biochemistry and Toxicology Lab. of RDDL (ER), Kolkata performed Biochemical Assay for 410 samples for the year 2015 -16 out of 665 samples received from West Bengal.

All the samples were tested against Total Protein, Albumin, Calcium and Phosphorus.

M. Soil testing of Anthrax Spore in the Endemic area (as per technical programme- 19) :

M-1. Screening of soil samples for B. anthracis :

Soil samples were received from different parts of the state & outside of the state from

the surface of the soil contaminated by the terminal discharge of the suspected animals or from endemic zone of anthrax and processed for isolation of B. anthracis spores as per the methods of OIE, 2004.

Ten soil samples were received from West Bengal and rest ninety six soil samples were received from outside state (Orissa). Out of 106 samples processed, nine soil samples from Odisha were found positive.

M-2. Details of soil sample processed :

Sl. No.

Source of Samples

No. of Samples Received

Positive

Negative

WEST BENGAL

1. Tufanganj-I, Coochbehar 1 0 1

2. Simlipal, Bankura 4 0 4

3. Kalna-Ii, Burdwan 1 0 1

4. Kalna-I, Burdwan 2 0 2

5. Puncha, Purulia 1 0 1

6. Jayrambati, Bankura 1 0 1 ODISHA

7. Udala, Rayagada, Simligura (Koraput) Kokokoraput

11 0 11

8. Bahalad, Mayurbhanj 1 0 1

9. Tumidhibandha, Kandhamal 10 1 9

10.

Udala&Khunta, Mayurbhanj 3 2 1

11.

Kapadanga, Raygada 15 0 15

12.

Simlipal, Mayurbhanj 18 0 18

13.

Lamtaput, Koraput 3 2 1

14.

Daspalla, Nayagarh 4 0 4

15.

Laxmipur, Koraput 7 3 4

16 Dasamantpur, Koraput 2 0 2

17 Baipariguda, Koraput 2 1 1

18 Dhenkanol 1 0 1

19 Budhimara, Mayurbhanj 3 0 3

20 Gudugudia, Mayurbhanj 9 0 9

21 Khuldia, Balasore 4 0 4

22 Dukuria, Baripada 1 0 1

23 Nawana, Baripada 1 0 1

24 Pandapada, Keonjhar 1 0 1

TOTAL 106 9 97

30

N. Testing of Export Materials (as per technical programme-20) :

A total 102 nos. of sample were received from different sources through Regional officer (ER) Animal Quarantine and Certification Services, NSCBI Airport, Kolkata for testing. All the samples were tested as per the methods of OIE using specific media. All of 102 no. of samples were negative to Bacillus anthracis.

O. Maintenance of repository of infective agents isolated from the Eastern Region (as per technical programme-21) :

Work has been undertaken to maintain different bacterial and viral agents in this laboratory as State Repository. Following infectious agents so far isolated and identified are maintained as state repository as per mandate of RDDL.

O-1. Maintenance of Bacterial agents :

A good no. (395) of bacterial isolates from different clinical and environmental samples are maintained by subculture in artificial media in this section as State Repository.

Sl. No.

Name of Bacterial Agent

Maintained by

Isolated from

Routine Subculture in Artificial

Media

No. of Isolate

1 Typed Escherichia coli -Do- 38 Poultry, Love bird, Goat, Pig, Rabbit, Garole Sheep etc.

2 Escherichia coli -Do- 112 Poultry, Pheasant, Black Swan, Badrika, Dog, Garole Sheep, Cocktail, Bovine, Ostrich, Kangaroo, Tiger, Lengur Rabbit, Spoonbill, Swamp Deer, Wild Elephant, etc.

3 Screened Escherichia coli -Do- 22 Poultry

4 Escherichia coli -Do- 11 Solely from wild animals

5 Typed Salmonella sp. - Do - 30 Poultry , Cat, bovine etc.

6 Salmonella sp. -Do- 45 Poultry, bovine, caprine, etc.

7 Proteus -Do- 17 Dog, poultry, Rabbit, etc.

8 Pasteurella multocida - Do - 16 Duck, Cattle , Buffalo, Poultry

9 Pasteurella haemolytica - Do - 01 Pin tailed duck

10 Bacillus anthracis - Do - 37 Cattle, Goat, Soil.

11 Pseudomonas sp. - Do - 23 Dog, Pig, Poultry.

12 Streptococcus sp. - Do - 6 Milk.

13 Staphylococcus sp. - Do - 31 Horse, Tiger, Deer, cattle etc.

14 Listeria monocytogenes - DO - 02 American Type Culture Collection (ATCC), Bovine etc.

15 Staphylococcus aureus (Cowan Strain)

- Do - 02 Microbial Type Culture Collection, Chandigarh.

16 Streptococcus agalactiae - Do- 01 ATCC

20 Salmonella pullorum St.-17 -Do- 01 Avian

TOTAL 395

31 O-2. Maintenance of Viral agents :

In our laboratory we are maintaining the following viral agents isolated from poultry and duck origin using Embryonated eggs.

Sl No. Name of Viral Agent Isolated from Maintained in

01 RDV Poultry Hens’ embryo

02 DPV Duck Ducks ’embryo

P. Organizing training programme for veterinary & para-veterinary work force (as per technical programme -22) : Training Programme on „Advanced Laboratory Technique for Screening of Sexually

Transmitted Diseases in Bovine‟ was held from 12-16 May, 2015 and „PCR Technique for Diagnosis of Common Bacterial, Viral & Parasitic Diseases of Livestock‟ was held from 23-27 February, 2016 in this Institute with the participating officers of different States under juricdiction of ERDDL.

Q. Preparation of different audio-visual presentation for mass-awareness of vets/para vets and common people among economically important livestock & poultry diseases (as per technical programme -23) :

No such activity was done in this Financial Year.

R. Detection of Microbial causes responsible for clinical and sub- clinical Mastitis and its anti-biogram (as per technical programme-24) :

To identify the early and latest cases of Mastitis and causal agent, Mastitis Control Unit of this Institute received/collected 205 samples from different organized and private farms. A total of 175 samples showed positive. The test results are depicted in the following table :

To

tal

sa

mp

le

rece

ive

d/ c

oll

ecte

d

Sa

mp

le s

ub

ject

to

scre

en

ing

Result of screening Organisms Isolated

Sta

ph

ylo

co

cc

us s

p.

Str

ep

toco

ccu

s s

p.

P.

ae

rug

ino

sa

E.

Co

li

Oth

ers

TO

TA

L

+ ve - ve

205 205 175 30 93

(53%) 14

(8%) 26

(15%) 42

(24%) - 175

32

S. Monitoring of the state sponsored schemes on Disease Investigation (as per technical programme-25) :

S-1. P.M Examination of different Poultry birds :

Month Duck Turkey Case birds

Pigeon

Fowl

Broiler Colour

Broiler*/ Kroiler

Cockrel RIR Layer

Red jungle Fowl

Vana- raja

Harin- Ghata- B.F.

ND*

April - - 5 4 75 - - - - - - - -

May - - 2 2 36 - - - - - - - -

June 2 - 1 - 37 - - - - - - - -

July 22 - - 2 56 - - - - - - - -

Aug. - - - - 26 - - 5 - - - - -

Sept. - - - - 66 - - - - - - - -

Oct. - - - - 77 - - - - - - - -

Nov. - - 3 1 51 - - 3 - - - - -

Dec. - - 10 - 66 - - 3 - - - - -

Jan. 2 - - 2 59 - - 1 - - - - 1

Feb. - - 1 6 45 - - 3 - - - - -

March - - 3 1 58 - - 4 - - - - -

Total 26 - 25 18 652 - - 19 - - 1

* Non-descriptive

S-2. P.M Examination of Small and Lab. animals :

Type of cases Small animals Laboratory animals

Month Pig Dog goat Rabbit Guinea-pig Mice

April - - - - - -

May - - - 4 8 -

June - - 1 5 2 -

July - - 2 - - -

August 1 - - 8 1 -

September - - 8 5 - -

October 1 - 2 - - -

November - 1 - - - -

December - - - - - -

January - - - - - 10

February - - - - 4 -

March - - - - - -

TOTAL 2 1 13 22 15 10

33 S-3. Necropsy (Post mortem) findings/Suggestive diseases of Poultry :

Type of P.M lesions Total

Brooder Pneumonia 49

Brooder Pneumonia/E. coli 5

Colibacillosis/ Colisepticaemia / Coli-granuloma 180

CRD / CCRD 23

Coccidiosis 62

Coccidiosis & RD 7

Colibacillosis & RD 12

Heat Stress / Severe Dehydration 7

Helminthiasis (Ascariasis, Capillariasis, Cestodeasis etc.) 5

IBD / Scl. IBD 39

Infectious Bronchitis (Cl. / Sub.Cl.) 59

Mycotoxicosis & Immunosuppression 5

Ranikhet Disease 54

Salmonellosis 45

Staphylococcus 11

Visceral Gout / Articular Gout 82

Duck Plague 22

Others 61

S-4. Wildlife samples for post-mortem examination:

Sl. No.

Type/Species of Wild animals /birds

PM done (Nos.)

P.M. and Lab. observation

WILD BIRDS

1 African Parrot 3 E.coli

2 Love Bird 12 Tape worm

3 Badrigar 3 Enteritis

4 Cockateil 5 NDV

5 Large Indian Parakeet 1 NDV

6 Java 1 NDV

Total 25

REPTILES

1 Tortoise 02 E.coli

34 S-5. Postmortem Materials & isolates sent/analyzed at different laboratories :

S-6. Month wise Samples Received for Laboratory Examination at BTTCU :

Sl. No.

Type of Material Sent/ Cultured

Name of the Section Agent Identified / Findings

(Report as received)

1.

PM Materials / Bacterial isolates / Impression Smear

& Blood Slide

Research Laboratory on Bacterial isease/Specialist ,

Diagnostic Bacteriology Lab., ERDDL

E.coli.- 195 Salmonella sp.- 43

Staphylococcus sp.- 14 Pasteurella sp. - 4 Clostridium sp. - 4

2. PM Materials RLBD Lab.* / Diag.

Virology Lab., ERDDL

New Castle Disease Virus (NDV) – 71 Infectious Bursal Disease Virus (IBDV) -28

Infectious Bronchitis Virus (IBV) – 45 Aspergillus - 29

Proteus - 55

3. Tissue materials

for Histopathology Diagnostic Pathology Lab.

Positive findings according to HP study : (a)Nephritis & Gout

(b) Cardiac myopathy (c) FLKS (d) Bursitis

(e) Encephalopathy (f) Mycotic pneumonia

4. Tissue materials HPLC Lab. Aflatoxin

5. Intestinal

scrapping/content Diagnostic Parasitology,

ERDDL

Eimeria sp.-134 Cappilaria sp.-2 Round worm - 4

Tape worm - 2

Month & Year

Positive cases

Total Blood slides Intestinal

scrapping Skin

scrap. Ecto-

parasite Whole Blood

Whole worm

April 4 18 - - - - 22

May - 8 - - - - 8

June 10 12 - - - - 22

July 42 15 1 12 1 - 71

August 12 8 - - - 11 31

September 34 16 3 - 1 - 54

October 28 12 - - - - 40

November 23 12 - - - - 35

December 8 11 - - - - 19

January 4 8 - - - 7 19

February 12 18 - - - - 30

March 13 23 - - 5 8 49

TOTAL 190 161 4 12 7 26 400

35

S-7. Type of Parasites identified at BTTCU :

(A) Haemoprotozoan Parasite (B) Eggs /Ova of Helminth

(C) Oocyst of Protozoa (D) Ticks/ Mites

(E) Whole worm

T. Monitoring of Centrally sponsored scheme like NPRE and State ELISA Laboratory (as per technical programme-26, 10) :

T-a. Report of Paste-Des-Petits Ruminants (PPR) Diagnostic Laboratory :

Established in 1997, the State ELISA Laboratory of Institute of Animal Health & Veterinary Biologicals (R&T) has been providing confirmatory diagnosis for Peste-des-Petits Ruminants (PPR), a highly infectious disease of sheep and goats. During the year 2015-2016 total 599 nos. samples were tested for antigen and 322 nos. samples were found positive. Samples were collected as Nasal Samples, Ocular Samples, Buccal Samples, Rectal Swab and Post Mortem materials.

Sl. No.

Haemoparasite Animal

Infected

1 Anaplasma sp. Cattle & Goat

2 Anaplasma sp. with Theileria annulata

cattle

3 Theileria annulata Cow (CBJ)

4 Babesia bigemina C/B Cattlle

5 Babesia gibsoni Dog

6 Microfilaria Cattle

Sl. No.

Name of Helminthic eggs/ova

Animal/ Birds Infected

1 Strongyle Cattle, sheep, goat

2 Paramphistomum sp. Cattle, Goat

3 Fasciola sp. Cattle

4 Monezia sp. Cattle, Goat

5 Trichuris sp. Goat, Rh. Monkey &

langur

6 Strongyloides sp. Cattle, Goat, Dog, Rh.

Monkey & langur

7 Toxocara vitulorum Cattle calf

8 Ancylostoma caninum Dog

9 Ascaridia galli Poultry

10 Echinococcus sp. Dog

11 Taenia sp. Dog

12 Diphylobothrium latum Dog

Sl.

No.

Name of Protozoan

Oocyst/ Cyst

Animal/ Birds

infected

1 Oocyst of Eimeria sp. Poultry, Cattle

Rabbit, Goat, Sheep

2 Cyst of Balantidium coli Cattle

Sl. No.

Name of Tick/ Mites Animal Infected

1 Boophilus microplus Cattle

2 Rhipicephalus sp. Cattle

Sl. No.

Name of Helminth Animal/ Birds Infected

1 Paramphistomum sp. Cattle

2 Rhabdias sp. (Round worm) Indian Rat Snake

3 Ophidascaris ajgaris (Round worm) Indian Rock Python (Moyal)

3 Oochoristica sp. (Tape worm) Land Monitor (Lizard)

4 Ascarid worm Indian Rock Python

5 Bothridium pythonis (pseudophyllidaen tape worm) Indian Rock Python

36

T.a-1. Diagnosis of Peste des Petits Ruminants (PPR) at a glance :

T. a-2. Outbreak & Samples Receipt Pattern of PPR in different Districts of West Bengal during 2015-16 :

Particulars No. No. of outbreaks for which samples turned positive to PPR 132

Total no. of samples tested for antigen 599 Total no. of samples found positive to PPR 322

Total no. of districts of West Bengal in which PPR was documented 15 Total no. of PPR samples received only in West Bengal 554

Total no. of samples found positive to PPR only in West Bengal 311 Total no. of suspected outbreak reported from West Bengal 158

Total no. of positive outbreak reported from West Bengal 126 Total no. of suspected outbreak reported from outside state 08 Total no. of positive outbreak reported from outside state 06

Total no. of serum samples screened for antibody to PPR virus 2448 Total no. of post vaccination serum samples screened for antibody

to PPR virus in West Bengal 1591

Total no. of post vaccination serum samples found positive for antibody to PPR virus in West Bengal

1257

Sl. No.

Name of Districts

No. of

samples

received

No. of

positive

samples

Suspected outbreaks

Positive outbreaks

1. Bankura 22 14 9 9

2. Birbhum 31 24 27 21

3. Burdwan 14 8 7 5

4. Cooch Behar No samples received

5. Dakshin Dinajpur No samples received

6. Darjeeling No samples received

7. Hooghly 24 22 9 9

8. Howrah 47 33 15 12

9. Jalpaiguri 10 6 6 5

10. Kolkata 11 2 2 1

11. Malda 21 17 8 4

12. Murshidabad 1

0

1 0 13. Nadia 78 34 11 11

14. North 24 Parganas 1 1 1 1

15. Paschim Medinipur 34 21 10 8

16. Purba Medinipur No samples received

17. Purulia 157 95 41 31

18. South 24 Parganas 5

3 2 2

19. Uttar Dinajpur 98 31 9 7

Total 554 311 158 126

37

T.a-3. Seasonal outbreak Pattern of PPR in the region during 2015-16 :

Month Suspected

PPR

Outbreaks

Positive Outbreaks

Within State

Outside state

April 4 4 0

May 13 10 0

June 23 21 1 (Odisha)

July 32 21 0

August 23 17 0

September 17 7 2 (Odisha)

October 4 1 1 (Odisha)

November 13 13 0

December 8 8 0

January 11 10 1 (Bihar)

February 8 5 1 (Odisha)

March 11 9 0

TOTAL 167 126 6

T.a-4. Seasonal Outbreak Pattern of PPR (confirmed) in West Bengal during 2015-16 :

The diagrammatic representation on the matter is as follows :

38

T.a-5. Type of Samples positive for PPR during 2015-16 :

Type of Samples

Positive Samples

Negative Samples

Total samples Percentage of

samples positive

Nasal Samples 270 273 543 49.72

Ocular Samples 1 3 4 25.00

Buccal Samples 11 2 13 84.61

Rectal Swab 8 8 16 50.00

Post Mortem materials

15 2 17 88.23

Whole Blood 6 0 6 100.00

Serum 39 19 58 67.24

TOTAL 350 296 657 53.27

T.a-6. Real Time RT-PCR Test Using Sybr Green for Diagnosis of PPR Infection

rRt-PCR(real time RT-PCR) using sybr green assay (melt curve analysis) has been performed in the year 2015-16 for extracted viral RNA from whole blood collected in EDTA by field units.

Melt peak analysis of PPRV M gene amplified products by RT –PCR sybr green assay

Sample Name Gene Name TM1 (°C) TM2 (°C)

Dye

PPRV POS PPRVM gene 86.02 - SYBR Green I

PPRVsample3 PPRVM gene 86.28 - SYBR Green I

PPRV POS PPRVM gene 85.95 - SYBR Green I

PPRVsample3 PPRVM gene 86.13 - SYBR Green I

PPRVsample1 PPRVM gene - 77.18 SYBR Green I

PPRVsample5 PPRVM gene - 77.19 SYBR Green I

PPRVsample1 PPRVM gene - 77.11 SYBR Green I

PPRVsample5 PPRVM gene - 77.13 SYBR Green I

PPPVsample2 PPRVM gene - 77.15 SYBR Green I

PPRVsample6 PPRVM gene - 77.14 SYBR Green I

PPPVsample2 PPRVM gene - 77.21 SYBR Green I

PPRVsample6 PPRVM gene - 77.13 SYBR Green I

NTC PPRVM gene - 77.20 SYBR Green I

NTC PPRVM gene - 77.18 SYBR Green I

NTC PPRVM gene - 77.19 SYBR Green I

NTC PPRVM gene - 77.17 SYBR Green I

39

T.b. National Project on Rinderpest Eradication work in West Bengal during 2015-16 :

T.c. Activity Report of AICRP on FMD Laboratory :

Technical Summary of the Project Work

The Laboratory of AICRP on FMD has been established in Kolkata with a view to the objectives are to conduct Epidemiological studies on Foot and Mouth disease in the state of West Bengal. These include geographical distribution, seasonal trend of different FMD Virus types associated with the disease and the various factors associated with origin, course and development of outbreaks including the incidence of FMD in organized farms, also monitoring the immunity status of animals where regular vaccination is undertaken. The Regional Centre performs the following Technical programme as per ICAR mandate:

(A) Epidemiological Studies ;

1. Number and distribution of various susceptible animals at risk. 2. Morbidity and Mortality pattern of different age group.

3. The seasonal trend of the disease and routes of migration of animals. 4. Studies on correlation between the outbreak of the disease and the migratory routes of the animals.

5. Determination of relationship of environmental factors viz. temperature, relative humidity and rainfall with the incidence of FMD outbreak.

6. Source and mode of spread of FMD outbreaks. 7. Estimation of economic losses due to the disease as per ICAR guidelines.

Name of District Village search target

Village search

achieved

Percentage achieved

Day book inspection

target

Day book inspection achieved

Percentage achieved

Bankura 3540 3540 100.00 1175 1102 93.80

Birbhum 2229 2229 100.00 1050 939 89.40

Burdwan 2570 2570 100.00 975 882 90.50

Cooch Behar 1139 1102 96.75 625 560 89.60

Dakshin Dinajpur 1486 1351 90.91 725 602 83.03

Darjeeling 656 520 79.27 525 411 78.28

Hooghly 1899 1899 100.00 975 975 100.00

Howrah 755 755 100.00 700 700 100.00

Jalpaiguri 736 698 94.84 675 602 89.18

Malda 1615 1615 100.00 925 925 100.00

Murshidabad 1927 1927 100.00 1100 1100 100.00

Nadia 1255 822 65.50 950 821 86.42

North 24 Pgs. 1619 1619 100.00 1100 1100 100.00

Paschim Medinipur 7360 7251 98.52 675 675 100.00

Purba Medinipur 3053 3042 99.64 925 925 100.00

Purulia 2452 2452 100.00 1100 1100 100.00

South 24 Pgs. 2125 1938 91.20 1100 898 81.64

Uttar Dinajpur 1550 1550 100.00 725 725 100.00

TOTAL 37966 36880 97.14 16025 15042 93.86

40

(B) Virological studies ;

1. Typing of FMD virus from collected field samples by Sandwich ELISA technique. 2. Sero-surveillance study by Liquid Phase Blocking (LPB) ELISA as a component of National FMD Sero-Surveillance Programme.

3. Sero-conversion study by LPB ELISA for West Bengal. 4. Assessment of quantitative sero-epidemiology from randomly collected sera samples. 5. Study of carrier status of infection and latency of the virus in bovines through detection of Anti-3AB3 NSP antibodies by DIVA (differentiation of infected from vaccinated animals) ELISA. 6. Regular transmission of collected field materials along with epidemiological data and test results to the Central FMD Lab., Mukteswar for genetic characterization of the virus.

T.c-1. FMD Outbreaks in West Bengal :

T.c-2. Sero monitoring under National Sero Surveillance Programme (Random samples)

No. of confirmed

outbreak

Type virus involved No. of samples

tested Type virus involved

O A C Asia-1

79

O A C Asia-1 No Virus

Detected

23 22 - - 01 53 - - 01 25

Name of the State

Species of Animals

Total No. of serum samples received (Prevac+ Postvac)

Total No. of serum samples tested

(Prevac+ Postvac)

West Bengal Cattle 3282 3282

Other States

- - -

TOTAL 3282 3282

41 T.c-3. Results of DIVA test of the samples randomly collected from different districts of West Bengal

U. Screening of Listeria sp. from bovine brain samples (as per technical programme- 9 ) :

A total 31 number of samples received through Specialist Pathologist, ERDDL,

Kolkata, which were tested for Listeria sp. The entire samples were processed after

cold enrichment and following standard methods. All the samples were found negative

for Listeria sp.

V.a. Other Virological work (including Molecular Biological Diagnosis) at a glance (as per technical programme-10)

Standardization of molecular biological diagnostic techniques for different pathogens of

livestock and poultry diseases other than Avian Influenza has been taken during the

year 2015-16.

Name of the State

Species of animals

Total no. of

sera samples received

for DIVA

Total no. of

sera samples

tested for DIVA

West Bengal Cattle 4023 4023

Other States

- - -

TOTAL 4023 4023

Type of Samples Source of Samples No. of Sample

Results

Positive Negative

Obex of Medulla of Bovine Brain

Specialist, Diagnostic Pathology, ERDDL, Kolkata

31 - 31

TOTAL 31 - 31

42

Sl. No.

Source of samples

Type of Samples Test Done No. of

Samples Tested

RESULT

Positive Negative

1. Bihar PM material PCR for detection of Avi Pox viral DNA 2 2 -

2. West Bengal Scab Material / Swab

PCR for detection of Capri Pox viral DNA 13 13 -

3 Tripura 7 7 -

4 West Bengal Brain Sample Rapid Immun Chromatographic tech. for detection of Rabies viral Antigen

2 1 1

5 West Bengal PM material

Immuno Chromatography for detection of IB/ND/IBD viral Ag.

470 153 317

6 Bihar 2 2 -

7 West Bengal PM material /

whole Blood

PCR for detection of Duck viral

enteritis (DVE) DNA

7 4 3

8 A & N Islands

84 - 84

9 A & N Islands

PM material / whole Blood

RT-PCR for detection of Duck viral hepatitis (DVH) RNA

50 - 50

10 West Bengal PM material / whole Blood

RT-PCR for detection of Classical Swine Fever viral RNA

3 3 -

11 Bihar 7 7 -

12 Sikkim 4 4 -

13 Tripura Swab PCR for detection of Lumpy Skin

Disease (LSD) viral DNA 8 - 8

14 West Bengal Scab

(from mouth/lip) PCR for detection of Contagious

Ecthyma viral DNA 11 11 -

15 West Bengal

Serum sample

(pre & post vaccinated)

AGPT for detection of Goat Pox

antibody in serum samples 96 46 50

TOTAL 766 253 513

V.b. Other Bacteriological work of Molecular Biological Diagnosis at a glance (as per technical programme-10) :

Different referral samples from different sources under jurisdiction of RDDL(ER) have been diagnosed during the year 2015-16. Moreover, Standardization of molecular biological diagnostic techniques for different pathogens of livestock diseases has also been taken during the year 2015-16.

Sl. No.

Source of samples

Type of Samples Test Done No. of Samples

tested

RESULT

Positive Negative

1. West Bengal PM material

PCR for detection of Mycobacterium tuberculosis

Genome

1 1 -

2. Bihar 1 1 -

3. Odisha PM material / whole Blood

PCR for detection of Bacillus anthracis

Genome 4 4 -

4. A & N Islands Nasal Swab

PCR for detection of Mycoplasma mycoides

Genome 339 37 302

5. West Bengal PM material /

whole Blood

PCR for detection of Pasteurella multocida

Genome 5 5 -

TOTAL 350 48 302

43

V.c. Other Parasitological work of Molecular Biological Diagnosis at a glance (as per technical programme-11) :

Standardization of molecular biological diagnostic techniques for different parasites of livestock has been taken during the year 2015-16.

Sl. No.

Source of samples

Type of Samples Test Done No. of Samples

Tested

RESULT

Positive Negative

1. West Bengal Whole blood

PCR for detecting genome of Theileria sp.

1 1 -

2. Sikkim 90 - 90

3. West Bengal Whole blood

PCR for detecting genome of Babesia bigemina

3 3 -

4. Sikkim 90 - 90

5. West Bengal

Whole blood PCR for detecting genome of

Anaplasma sp.

6 6 -

6. Sikkim 90 - 90

7. Odisha 1 - 1

TOTAL 281 10 271

W. Physical achievement of ERDDL for the year 2015-16 at a glance:

Sl. No.

Source of the samples No. of samples

received No. of samples

tested

R E S U L T R e m a r k s

Positive Negative

VIROLOGICAL SAMPLES UNDER SPECIALIST, DIAGNOSTIC VIROLOGY

1 West Bengal 31,860 31,860 708 31,152

2 Bihar 5,205 5,205 13 5,192

3 Odisha 7,864 7,864 9 7,855

4 A & N Islands 302 302 0 302

5 Sikkim 38 38 11 27

6 Tripura 2,121 2,121 7 2,114

TOTAL 47,390 47,390 748 46642

SEROLOGICAL SAMPLES UNDER SPECIALIST, SERO-EPIDEMIOLOGY

1 West Bengal 30,907 30,907 4,898 26,009

2 Bihar 2,886 2,886 Nil 2,886

3 Odisha 2,777 2,777 208 2,569

4 A & N Islands 170 170 0 170

5 Sikkim 5 5 0 5

6 Tripura 1,506 1,506 0 1,506

TOTAL 38,251 38,251 5,106 33,145

BACTERIOLOGICAL SAMPLES UNDER SPECIALIST, DIAGNOSTIC BACTERIOLOGY & MYCOLOGY

1 West Bengal 2,181 2,181 592 1,589

2 Bihar 42 42 2 40

3 Odisha 410 410 17 393

4 A & N Islands 341 341 37 304

5 Sikkim 7 7 0 7

TOTAL 2981 2981 648 2,333

PARASITOLOGICAL SAMPLES UNDER SPECIALIST, DIAGNOSTIC PARASITOLOGY

1 West Bengal 2,120 2,120 965 1,155

2 Bihar 24 24 0 24

3 Odisha 113 113 1 112

4 Sikkim 91 91 0 91

TOTAL 2,348 2,348 966 1,382

44

BIOCHEMICAL & TOXICOLOGICAL SAMPLES UNDER

SPECIALIST, DIAGNOSTIC BIOCHEMISTRY & TOXICOLOGY

Sl. No.

Source of the samples

No. of samples received

No. of samples tested

R E S U L T R e m a r k s

Positive Negative

1 West Bengal 678 422 53 * 357#

* beyond the reference value # within the normal value

2 Bihar 4 - - -

TOTAL 682 422 53 357

PATHOLOGICALS SAMPLES UNDER SPECIALIST, DIAGNOSTIC PATHOLOGY

1 West Bengal 20 *+80** 20 +80 0 20+80 *BSE-Brain

Samples from clinical suspected cases **BSE- Brain Samples collected from slaughter house # other than BSE Samples

70 # 63 - -

2 Bihar 10** 10 0 10

4# 4 - -

3 Odisha 23* 23 0 23

4 A & N Islands 6* 6 0 6

5 Sikkim 5* 5 0 5

6 Tripura 6* 6 0 6

TOTAL 60*+90**+74#

= 224 217 - 60*+90**=150

GLIMPSE OF SAMPLES HANDLED AT RDDL, ER, KOLKATA DURING 2015 – 16

Sl. No.

Source of the samples

No. of samples received

No. of samples tested

R E S U L T

Positive Negative

1 West Bengal 67,916 67,653 7216 60,362

2 Bihar 8,175 8,171 15 8,152

3 Odisha 11,187 11,187 235 10,952

4 A & N Islands 819 819 37 782

5 Sikkim 146 146 11 135

6 Tripura 3,633 3,633 7 3,626

91,876 91,609 7,521 84,009

DETAILS OF SAMPLES TESTED IN RDDL, ER, KOLKATA DURING THE LAST 6

FINANCIAL YEARS

Sl.

No.

Financial Year Number of

Samples

Tested

1 2010-11 69240

2 2011-12 43796

3 2012-13 42406

4 2013-14 57448

5 2014-15 73530

6 2015-16 91609

45

ANNUAL ACTION PLAN AND TECHNICAL PROGRAMME

FOR RDDL (ER), KOLKATA

2016 - 2017

1. To provide referral and consultancy services to the state diagnostic laboratories for

economically important Livestock and poultry diseases.

2. Development of epidemiological Data Bank including epidemiological maps of

economically important livestock diseases of the region.

3. Screening of Bulls, teaser animals and bovine semen against sexually transmitted

diseases including estimation of microbial load in Frozen Semen Straw in the region.

4. qPCR based diagnosis with special reference to detection BoHV-1 in bovine semen

sample.

5. Surveillance work of infectious causes of abortion in Livestock population in

organized farm and in the rural areas.

6. Surveillance work on BSE based on Histopathological studies and ELISA based

diagnosis of brain samples (as per target of Govt. of India).

7. Screening of brain samples from Bovine Sp. exhibiting neurological disorders for

Listeriosis.

8. Immune status studies on Ranikhet Disease in vaccinated organized and backyard

poultry population.

9. Application of molecular biological techniques for routine diagnosis of different

livestock & poultry diseases (other than Avian Influenza) with special reference to

Capri pox, PPR, Classical Swine Fever, Duck Viral Enteritis, Duck Viral Hepatitis, New

Castle Disease, Brucellosis, Listeriosis,. Anthrax, HS, CCPP, Tuberculosis. etc.

10. Routine application of PCR techniques for diagnosis of haemoprotozoan infection in

cattles like Babesiosis, Anaplasmosis,and Theileriosis etc.

11. Routine Sero-surveillance work and antigen detection in swab / tissue samples of

Avian Influenza as per guideline of Govt. of India and framed surveillance strategies

adopted by different states under jurisdiction of RDDL (ER), Kolkata.

46

12. Diagnosis of Rabies infection in post-mortem material by immuno-chromatographic

technique.

13. Proximate Analysis of animal and poultry feed.

14. To impart analytical facilities of pesticides, antibiotics and drug residues in livestock

and poultry products by HPLC.

15. A) Study of nutritional status in infertility cases.

B) Clinical Haematology and Bio-chemistry to aid disease diagnosis.

16. Soil testing of Anthrax spore in the endemic area.

17. Testing of export materials as per OIE protocol.

18. Maintenance of Repository of infective agents isolated from the Eastern Region.

19. Organizing training programme for veterinary & para-veterinary work force.

20. Detection of microbial agents responsible for clinical and sub-clinical mastitis and its

antibiogram.

21. Monitoring of State Sponsored Scheme on disease investigation.

26. Monitoring of Centrally Sponsored Scheme of NPRE/AICRP on FMD.

27. Surveillance on Tuberculosis and Johne‟s disease by Tuberculin / Johnin Sensitivity

Test.