DBT-NER-PROJECT ADVANCED ANIMAL DISEASE DIAGNOSIS AND MANAGEMENT CONSORTIUM. Roadmap,

DBT-NER-PROJECT ADVANCED ANIMAL DISEASE DIAGNOSIS AND MANAGEMENT CONSORTIUM Roadmap, 2014-16 After a threadbare discussion and interaction among the members, a consensus was reached to prepare a road map with common minimum programme so that a visible progress can be achieved. Identified areas are as follows: 1. Prioritization of livestock species to be covered in the first phase: Based on the priority of the North Eastern Region, diagnostic facilities will be created in core labs with the help of national laboratories for diseases of pigs, goats and ducks. Intensive investigation on diseases affecting these species will be carried out in NE states during the first two years. However, diagnostic activities on other animals will also continue at the same time. 2. Infectious agents to be studied : Infectious agents affecting pigs, goats and ducks are grouped as endemic and emerging/re-emerging. Animal species Infectious agent Endemic Emerging/Reemerging Bacteria Salmonella, Pasteurella, Erysipelas, Oedema disease, Carbapenem resistant E Coli. Anthrax; Clostridium, Brucella Carbapenem resistant E Coli. Virus CSFV, PPV, RV, JEV, Rota, PRRS Circo virus, PEDV, Swine Influenza, Pigs Nipah virus, TGEV, Goats Swine Pox, TTV Parasites Nematodes ( Ascaris, oesophagostomum, stephanurus, gnathostoma, capillaria etc), trematodes ( Fasciola, opisthorchis, artyfechinostomum, gastrodiscoides), Larval cestode ( Cysticercus cellulosae, hydatids), Haemoprotozoa, coccidia Bacteria Pasteurella, Clostridia, Listeria, Mycobacterium paratuberculosis, Mycoplasma, Anthrax, Brucella Virus PPR, Orf, BTV,Goat pox, Para influenza, Nipah,Encephalities Parasites Trematodes (Fasciola, Amphistome, Schistosomes), GI Nematodes (Haemonchus, mecistocirrus, oesophagostomum, bunostomum, cooperia), Cestodes (Moniezia, stilesia, avitellina, Taenia & Echinococcus), coccidian and haemoparasites (Babesia, theileria, anaplasma). causing virus

Ducks Bacteria Pasteurella, Salmonella, Botulism, Mycotoxin Virus DPV DV Hepatities, AI Parasites Trematodes (Echinostoma, tracheophilus, prosthogonimus), Nematodes ( Ascaridia, heterakis, porocaecum, tetramers), Cestode (Raillietina, Fibbriaria, hymenolepis), coccidi and haemoparasite. Each national partner and NE core lab will take responsibility to develop diagnostics and confirm diseases prevalent in eight states. Action to be taken by the respective centre as assigned below: Action to be taken by : Animal species Endemic & emerging/re-emerging infectious agents Bacteria Brucella, AMR Pigs Virus CSFV, PRRS,TTV Goats Bacteria Brucella, AMR, Mycoplasma Action to be taken by NIHSAD: Animal species Endemic & emerging/re-emerging infectious agents Pigs Virus Nipah, Ebola, Influenza A virus, PRRS Goats *Virus CCHF, NSD, RVF Ducks Virus Influenza A virus Action to be taken by NRCE/VTCC : Animal species Pigs Goats Bacteria Virus Parasites Virus Parasites Endemic & emerging/re-emerging infectious agents Salmonella, Pasteurella, Erysipelas RV, JEV, Swine Influenza, Trypanosoma (rest of the component to be taken up at Khanapara and NRCE will assist for other parasites) Para influenza, Encephalities causing virus Trypanosome (rest of the component to be taken up at Khanapara and NRCE will assist for other parasites) Action to be taken by NRC on pigs: Animal species Bacteria Endemic & emerging/re-emerging infectious agents Salmonella, Pasteurella, Brucella

Pigs Virus CSFV, RV, Circo, Parvo virus Action to be taken by Core Lab I : Animal Endemic & emerging/re-emerging infectious agents species Bacteria Salmonella, Pasteurella, Brucella, Erysipelas, AMR, Anthrax; clostridium,oedema disease Virus CSFV, PPV, RV,PRRS, Swine Pox. Pigs Parasites, vector Nematodes ( Ascaris, oesophagostomum, stephanurus, gnathostoma,capillaria etc), trematodes ( Fasciola, opisthorchis, artyfechinostomum, gastrodiscoides), Larval cestode ( Cysticercus cellulosae, hydatids), Haemoprotozoa, coccidian, vector Goats Bacteria Brucella, AMR, Pasteurella, Clostridia, Listeria, Mycobacterium paratuberculosis, Anthrax Virus Orf Parasites, vector Trematodes (Fasciola, Amphistome, Schistosomes ), GI Nematodes (Haemonchus, mecistocirrus, oesophagostomum, bunostomum, cooperia), Cestodes (Moniezia, stilesia, avitellina, Taenia & Echinococcus), coccidian and haemoparasites (Babesia, theileria, anaplasma). vector Bacteria Pasteurella, Salmonella, Botulism, Mycotoxin. Virus DPV Ducks Parasites Trematodes (Echinostoma, tracheophilus, prosthogonimus), Nematodes ( Ascaridia, heterakis, porocaecum, tetramers), Cestode (Raillietina, Fibbriaria, hymenolepis), coccidi and haemoparasite. Action to be taken by Core Lab II : Animal Endemic & emerging/re-emerging infectious agents species Bacteria Salmonella, Erysipelas, Brucella, AMR, Virus CSFV, JEV, Circo virus, PRRS Parasites, Nematodes ( Ascaris, oesophagostomum, stephanurus, Pigs vector gnathostoma,capillaria etc), trematodes ( Fasciola, opisthorchis, artyfechinostomum, gastrodiscoides), Larval cestode ( Cysticercus cellulosae, hydatids), Haemoprotozoa, coccidian, vector Goats Bacteria Brucella, AMR, Listeria Virus Nipah Parasites, vector Trematodes (Fasciola, Amphistome, Schistosomes ), GI Nematodes (Haemonchus, mecistocirrus, oesophagostomum, bunostomum, cooperia), Cestodes (Moniezia, stilesia, avitellina, Taenia & Echinococcus), coccidian and haemoparasites (Babesia, theileria, anaplasma), vector Bacteria Pasteurella, Salmonella, Afla toxin

Virus DPV Ducks Parasites Trematodes (Echinostoma, tracheophilus, prosthogonimus), Nematodes ( Ascaridia, heterakis, porocaecum, tetramers), Cestode (Raillietina, Fibbriaria, hymenolepis), coccidia and haemoparasite. Action to be taken by Core Lab III : Animal species Endemic & emerging/re-emerging infectious agents Bacteria Salmonella, Pasteurella, Brucella, AMR, E Coli., Oedema disease Pigs Virus PRRS Parasites, vector Nematodes ( Ascaris, oesophagostomum, stephanurus, gnathostoma,capillaria etc), trematodes ( Fasciola, opisthorchis, artyfechinostomum, gastrodiscoides), Larval cestode ( Cysticercus cellulosae, hydatids), Haemoprotozoa, coccidian, vector Goats Bacteria Brucella, AMR, Pasteurella, Listeria, Mycobacterium paratuberculosis Parasites, Vector Trematodes (Fasciola, Amphistome, Schistosomes ), GI Nematodes (Haemonchus, mecistocirrus, oesophagostomum, bunostomum, cooperia), Cestodes (Moniezia, stilesia, avitellina, Taenia & Echinococcus), coccidian and haemoparasites (Babesia, theileria, anaplasma), vector Bacteria Pasteurella, Salmonella, Afla toxin Ducks Parasites Trematodes (Echinostoma, tracheophilus, prosthogonimus), Nematodes ( Ascaridia, heterakis, porocaecum, tetramers), Cestode (Raillietina, Fibbriaria, hymenolepis), coccidi and haemoparasite. *Goat diseases on PPR, BTV, Goat pox will be covered from 3 rd year onwards as there are DBT funded ongoing projects on these agents. 3. Strengthening of Human Resource : The prime objective of the project is to develop disease diagnostic competency in NER Labs through training human resources. Accordingly certain topics are identified and decided to carry out training programmes at national as well as at core lab levels within second year of the project as per tentative time schedule indicated below: Training Module Training Details Tentative date Action to be taken by Trainees Passive survey Survey strategy, data entry, record keeping 22-24 April, 2015 NE core I/ Lab- Core lab personnels,

Active survey TADs samplings Sampling strategy, collection of samples, preservation & dispatching Lab, sample handing, advanced diagnosis of TADs state Animal Husbandry personnels April, 2015 NIHSAD Core lab personnels, SAHD personnels Field diagnosis Basic Microbial, pathological, parasitic methods Gross diagnosis, Microscopical, Serodiagosis, June, 2015 NE core Labs I/II/III state Animal Husbandry personnels Advanced diagnosis Molecular diagnostic protocols (OIE/Inhouse), Serodiagnosis, isolation techniques July, 2015 / NIHSAD/NRC E/VTCC/NRC pigs NE core Labs// HSADL/NRC E/VTCC/NRC pigs Disease mapping Software management & Data analysis GIS application, analysis and interpretation Data analysis, data interpretation & presentation May, 2015 / NE core Labs/ Core lab personals, state Animal Husbandry personals Reposition Methods on reposition & storage, Sample coding, record keeping, preservation, dispatch. August, 2015 NRCE/ VTCC Training carried out at National labs, necessary TA/DA for SAHD officers should be arranged by the host institutes and at core labs fund should be arranged by core lab. However, matter should be taken up at DBT level to obtain permission for payment of TA & DA from Training & also allotment of more fund under head Training. 4. Development of Laboratory capability at Core Labs : a. Protocols standardized/developed as per OIE guidelines and diagnostic kits/reagents/ positive control/negative control available at national labs as well as in other core labs should be shared with NE core labs. Feed back of test results can be used as validation of the tests. Such protocols can be finalised as common SOPs for use in the diagnosis of

diseases in NE states. Necessary MTA should be completed by respective National partners so as to dispatch kits/reagents to respective centres within 2 months. Agents PRRSV Type of SOPs/kits Action to be taken by available PCR protocol NIHSAD, Core lab II, SOPs/kits provided to Core lab I & III Indirect ELISA kits NISAD Core labs I,II,III JEV PCR protocol NRCE, Core lab II Core lab I & III HI protocol/kits NRCE Core labs I,II,III AIV Ag Ab kits NIHSAD Core labs I,II,III, Field labs NDV PCR protocol Core lab I Core labs II,III, HI Field labs protocol/reagents DPV PCR protocol Core lab I Core lab II, III i-elisa protocol RV PAGE, PCR protocol Swine pox Isolation protocol Core lab I, NRCE Core lab I Core labs II,III PCR protocol Core lab I Core labs II,III Fowl pox PCR protocol Core lab I Core labs II,III Swine influenza PPR, BTV, Goat Pox PCR protocol NRCE Core labs I,II,III PCR protocol Core labs I,II,III AMR PCR protocol, Core lab III Core labs I,II Brucella PCR protocol Core labs I,II,III, Ab kits Field labs Clostridia PCR protocol Core lab I Core labsii,iii, Isolation protocol National lab Listeria PCR protocol Core lab II Core labs I, III Trypenos oma Isolation protocol PCR protocol NRCE Core labs I,II,III, Field labs ELISA protocol

Babesia, Theileria PCR protocol Core lab I Core labs II,III, National lab b. Common protocols used by Core lab I, II and for detection of CSFV by PCR and Ag-Ab detection tests and isolation of the virus will be validated by ring tests. Consensus protocols in the light of EU SOPs will be finalized and developed as common SOP for use in diagnosis of CSFV in NE states. Action will be taken by Core lab I and. c. For development of SOPs to detect PPV, Circo virus, PEDV, TGEV, Nipah virus, TTV, Duck hepatitis virus, Erysipelas, Oedema disease, cysticarcus, hydatid, coenurus, qpcr for Theileria, Babesia,, Aflatoxin, Anthrax should be initiated by identified centres and necessary clinical/tissue samples should be provided from field by core labs for standardization of tests. 5. Development of Laboratory Capability at Field units : a. A diagnostic laboratory manual will be prepared covering necessary formats, protocols for preparation of various reagents, diagnosis of microbial, parasitic, toxicological diseases and clinic-pathological, pathological investigations etc. Action to be taken by scientists from, VTCC, Core labs I, II and III (2 months). b. All 3 core labs should arrange training for field staffs for collection of disease data, samples and despatch of samples to the core labs. Necessary reagents, plastic and glass wares, preservation/transport media should be provided. To begin with diagnostic activities quick and handy diagnostic kits/reagents should be arranged. 6. Formats to be prepared : Type of formats Passive survey format, outbreak survey, format for Sero- survey, routine parasitic survey, check list for collection of sample. Uniform sample code for tissue, sera, isolates will be developed. Guidelines & formats for reporting of investigation results for bacteria/virus/serologcal/ parasitic/toxic/pathological investigations. Sample entry & sample registry formats Disease data base/ Accessibility to NADRES. Development of Website Action to be taken (One month) VTCC ; ; core labs VTCC ; ; core labs NRCE ;VTCC ; core labs Core labs I, II and

Monthly report format from field unit 7. Sampling Strategy : a. Initially for 6 months survey will be confined in and around field diagnostic units of the respective state as there are no funds available in the 1 st year for SAHDs. Information on number of villages, type of species available, No of animal herd/unit/bird unit, Av animals per herd/unit, Av. birds per unit will be collected and intimated to through core Labs. Action to be taken by PIs of SAHD and Core lab I (31 st March 2015). b. will identify the village and framed the sample number, frequency for sera samples, clinical samples, faecal samples in routine survey as well as during outbreak. Action to be taken by (April 2015). c. Samples will be collected as stated below jointly by project staffs and Field lab incharges. All consumables/ medicines/vaccines to be used in field dung collection of samples should be arranged by core labs. Animal/bird Type of sample No. of sample/state Pig Goat Duck Sera sample (3month-1 yr age group) Blood smears (Haemoprotozoa) Vector survey Nasal swabs Faecal samples (herd/unit basis of young, grower, adult) Organs in dead carcass Sera sample (3month-1 yr age group) Blood smears (Haemoprotozoa) Vector survey Nasal swabs Faecal samples (herd/unit basis of young, grower, adult) Organs in dead carcass Sera sample (3month-1 yr age group) Blood smears (Haemoprotozoa) Tracheal swabs 200 (50 /F. Lab) 100 (25 / F. Lab) 100 (25 / F. Lab) 400 (100 / F. Lab) As per the mortality/ outbreak 400 (100 /F. Lab) 100 (25 / F. Lab) 200 (50 / F. Lab) 400 (100 / F. Lab) As per the mortality/ outbreak 100 (25 /F. Lab) 40 (10 / F. Lab) 40 (10 / F. Lab)

Faecal droopings (unit basis of young, grower, adult) Organs in dead carcass 200 (50 / F. Lab) Dead/ ailing bird d. The participation of NERDDL is crucial and agreement regarding sample sharing, diagnostic protocol implementation and disease informatics need to be worked upon. e. Samples collected in field will be dispatched to core labs through project staffs, messenger from field, through courier service /overnight service. Expenditure will be borne by respective core lab. Between core labs samples will be transported by project staffs/courier service at every month. Samples from core labs to national labs will be dispatched at every 3/6months by courier/messenger. Emergency samples will be sent by messenger. All cost involve in dispatching samples to national labs should be arranged by respective national lab. Official permission for interstate transport of samples / dispatch to national lab will be arranged by PIs of SAHD and National labs. f. A nodal liaison representative at the National Institute level needs to be finalised and communicated to all implementing partners. g. There should be communication of fortnightly reports regarding work progress under this project to Core lab I in order to implement a real time monitoring and evaluate progress achieved by the implementing partners.

1 ROAD MAP OF THE PROJECT FOR THE YEAR 2016-19 AND ACTION TO BE TAKEN : 1. Prioritization of diseases:based on the infrastructure available for disease diagnosis in the North Eastern Regionthe diseases need to prioritized using retrospective disease data of last 10 year for species wise prevalent in the region. 2. Development of diagnostics:based on met analysis, diagnostics need be developedfor the diseases prevalent in the region for respective species with the help of national partners. 3. Surveillance of Emerging/Re-emerging diseases:-be undertaken in a holistic way using proper sampling & diseases of regional importance. 4. Research/Actions to be undertaken by each institute: Each national partner and NE core lab will take responsibility to develop diagnostics and confirm diseases prevalent in eight states. Action to be taken by the respective centre as assigned below: a., Bangaluru : Molecular epidemiology & phylogenic analysis of various diseases including risk & met analysis. Development of fore-warning models with impact analysis of identified diseases of national importance. b. NIHSAD, Bhopal: Human resource development & capacity building of officials of NER to handle emerging /exotic diseases in the region.. Molecular genomics& diagnosis of trans-boundary/exotic diseases including phylogenic analysis. c. NRCE/VTCC, Hisar : Molecular genomics& diagnosis of equine diseases except influenza virus including phylogenic analysis & risk assessment. Proper maintenance of isolates received from other units with cataloguing. d. NRC on pigs, Guwahati Development of infrastructure for proper conventional & molecular diagnosis of pig bacterial diseases.

2 e. Core Lab I, II & III : Development of infrastructure for proper conventional & molecular diagnosis including molecular epidemiological studies on different livestock & poultry diseases in the region with their prioritization Development of competency & core competence in handling different diseases of regional importance including emerging & exotic diseases, Fore-warning disease modelling, risk assessment & impact analysis in cooperation with, 5. Diagnostics or Vaccine development:- It should be limited to one or two labs & in case any institution is developing any diagnostic/vaccine need be validated in other labs & that institution would be responsible for its commercialization. SOP s already developed need be standardized as per OIE/WHO norms & guidelines. Any new SOP developed by any institution need be validated & standardized & Core Labs be involved in such procedures. 6. Strengthening of Human Resource: Developdisease diagnostic competency in NER Labs through training human resources in following training programmes. Name of Training Lab, sample handing, advanced diagnosis of TADs Molecular diagnostic protocols (OIE/Inhouse), Serodiagnosis, isolation techniques GIS application, analysis and interpretation Data analysis, data interpretation & presentation Methods on reposition & storage, Sample coding, record keeping, preservation, dispatch. Name of Institution imparting training NIHSAD / NIHSAD/NRCE/VTC C/NRC pigs / NE core Labs/ NRCE/ VTCC Name of Labs receingtrainig Core lab &, SAHD personnel NE core Labs//HSADL/NRCE/V TCC/NRC pigs Core lab personals, state Animal Husbandry personals

3 6. Development of Laboratory Capability at Field units : a. Point of care diagnostic developed by National Labs, Core Labs should made available at field labs. Besides other conventional diagnostics tools and reagents should be procured by respective core labs and distributed to field labs. b. A diagnostic laboratory manual/protocols be made available on website. c. Training for field staffs will continue. d. Scientific staffs from respective core labs/ national labs visit field labs in convenient intervals and perform diagnosis in the field labs along with field staff. 7. Formats to be prepared : Type of formats Passive survey format, outbreak survey, format for Serosurvey, routine parasitic survey, check list for collection of sample. Uniform sample code for tissue, sera, isolates will be developed. Guidelines&formats forreporting of investigation results for bacteria/virus/serologcal/parasitic/toxic/pathological investigations. Sample entry &sample registry formats Disease data base/ Accessibility to NADRES. Action to be taken VTCC ; ; core labs VTCC ; ; core labs NRCE ;VTCC ; core labs Development of Website Core labs I, II and Monthly report format from field unit These formats should be evaluated through an expert committee &cross validations need be done to maintain uniformity and updating the same. 8. Sampling Strategy & Disease Risk Analysis: a. Risk analysis and KAP formats should be prepared by in association with Core Labs b. Field study should be carried out as routine disease survey and outbreaks investigation. c. Villages surrounding to the Field labs should be identified. will identify the village and framed the sample number, frequency for sera samples, clinical samples, faecal samples in routine survey as well as during outbreak. Along with routine survey data on Risk analysis and KAP will be collected.

4 d. Awareness & animal health camps should be organized in identified strategic locations to understand farmers problems and collect samples. Awareness through different media & distribution of leaflets in vernacular/hindi language. e. Samples will be collected as stated below jointly by project staffs and Field lab incharges. All consumables/ medicines/vaccines to be used in field dung collection of samples should be arranged by core labs. 9. Sharing of Samples & Data:Needs to be continued. a. To compile disease map linkage should be established with ongoing disease related projects under DBT, ICAR, DSTin NER to share samples/data. b. The participation of NERDDL is crucial and agreement regarding sample sharing, diagnostic protocol implementation and disease informatics need to be worked upon. c. Samples collected in field will be dispatched to core labs through project staffs, messenger from field, through courier service /overnight service. Expenditure will be borne by respective core lab. Between core labs samples will be transported by project staffs/courier service at every month. Samples from core labs to national labs will be dispatched at every 3/6months by courier/messenger. Emergency samples will be sent by messenger. All cost involve in dispatching samples to national labs should be arranged by respective national lab. Official permission for interstate transport of samples / dispatch to national lab will be arranged by PIs of SAHD and National labs. 10. Strengthening of National Repository.It should be continued & national repository at VTCC need be strengthened by sharing the samples & it would not be proper to develop another facility of same kinds due to involvement of huge manpower & financial implications.