Shrikrishna Isloor

A network for animal rabies diagnostic laboratories in India under OIE Twinning Programme: A success story

OIE twinned CVA-KVAFSU Rabies Diagnosis Laboratory ,

Dept.of Microbiology, Veterinary college, KVAFSU,

Hebbal, Bengaluru

Rabies: Endemic in India except for the Union Territories of Anadaman and Nicobar and Lakshwadeep.

WHO-APCRI Indian multicentric rabies

survey in 2017, laboratory based

surveillance for the status of rabies in

dogs / cats was carried out in the

islands of Andaman/ Nicobar and Lakshadweep.

So far, rabies positivity not detected in AN and Lakshadweep

Pathogenesis

3

• Highly neurotropic

• Incubation period could

vary

• Virus excreted through

saliva prior to symptoms

• Virus excreted

intermittently through saliva

1. Prior to 2013: Mostly Seller’s staining

Progress in Diagnostic systems for animal Rabies –

2. CVA-KVAFSU Rabies Diagnosis Lab. 2013:

3. OIE Twinning programme - 2016: Strengthening of diagnosis of rabies in India

Virus

Vaccinal

antibodies

Direct fluorescent antibody assay (DFA)

Positives

Negative

Direct Rapid Immunohistochemistry Test (dRIT) For Detection of Rabies Virus Antigen

The World organization for animal Health (OIE) Laboratory Twinning Programme

• A global programme

• OIE reference centres build expertise for important animal diseases

• Priority regions identified through twinning with national laboratories

• Improve the global capacity for disease detection, prevention and control through better governance of veterinary services.

Network development through National level Workshops

OIE TRAININGS/ WORKSHOPS

First Workshop

Sample receipt/collection /

registration / storage and recording

Second workshop

dRIT-DFA-Nested PCR training

Fourth workshop

Brain sample collection :

Conventional V/s Occipital foramen approach

CDC, Atlanta APHA, UK

Networking through workshops conducted in other states of India

Aug.2019

Outcome:

Training:

14 states

Samples:

17 states &

2 UTs.

Andaman &

Nicobar islands Lakshadweep

islands

Network (animal rabies

diagnosis in India)

Field diagnosis by trainees…

Theerthalli,

Shivamogga, Karnataka

Bidar, Karnataka

Guwahati, Assam

Bangalore Rural

Impact of Social Media – Brain sample collection: Jan. 2019

25th July 2019

DFA based detection of rabies viral antigen in animals: Jan 2012 to 31 May 2019

80 % of brain samples tested are from dogs; 66% of dog samples were positives

DFA based detection of rabies viral antigen in animals: Jan 2012 to 31 May 2019

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Negative

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Lab. Surveillance (RDL, VCH): Rabies positive cases in Bangalore (2014-2019)

Year Total tested No. Pos. No. Neg % pos.

2014 72 54 18 75

2015 102 69 33 68

2016 61 35 26 57

2017 99 69 30 70

2018 177 115 62 65

2019 (upto

June)

47 36 11 77

Total tested (2014 – 2018):558

Total positives : 378

Percent Pos. : 68

Real-Time PCR: LN 34 assay

• A multi-site evaluation of the LN34 assay in 14 laboratories. A total of 2,978 samples (1,049 DFA positive) from the Americas, Europe, Africa and Asia were tested.

• High sensitivity (99.90%), specificity (99.68%) compared to DFA

• Detects viral RNA in fresh, frozen, archived, deteriorated and formalin-fixed brain tissue. (Crystal et al., 2018)

G-L intergenic sequence

G- Gene N- Gene

Molecular characterization: Conventional PCR: N,G gene PCR didn’t reveal genetic diversity

WHO-APCRI survey on Rabies in India- Island survey in 2017

For the first time cat brain samples from Lakshadweep and dog brain samples from Andaman

tested and found negative so far

Control of rabies in India: a Veterinary perspective

• A National Rabies Control Programme (Zoonosis Division) was implemented in the 12th Five Year Plan of the Government of India

• The program has two components: Human and animal component.

• The strategy for the animal component:

* Population survey of dogs

* Dog population management

* Mass vaccination of dogs.

Enhancement of immune surveillance through sero-monitoring of rabies vaccinal antibodies in dogs

• Sero monitoring of vaccinal antibodies.

• RFFIT / FAVN /ELISA (limited facilities to conduct RFFIT )

• Need to adopt in Asian countries

ELISA for seromonitoring

PLATELIA TM RABIES ll ASSAY ELISA kit

Positive by

RFFIT

Positive by

ELISA

Negative by

RFFIT

Negative by

ELISA

42 40 8 10

Development of a quantitative ELISA for sero monitoring Post-anti rabies vaccinal antibody responses in dogs - a DBT funded project

The mass vaccination of dogs: a sustainable approach Human and Dog Rabies Cases, Mexico 1990-2006

MASS VACCINATION IN DOGS v/s HUMAN RABIES CASES

Dog vaccination and Human Rabies (Srilanka) Death rate/100,000 Pop.

Mass dog vaccination in Goa

Dog vaccinations and Human Deaths in Goa

Assessing vaccination and laboratory surveillance- Bengaluru (2018)

N= 250 Protected Unprotected

Number of

sample 125 125

Percentage 50 50

Mass Anti-Rabies Vaccination of dogs in BBMP jurisdiction – proposed pilot study in 2019

Components of campaign:

1. Stray Dog population survey

2. Mass Anti- Rabies Vaccination

3. Sterilization of female Stray-dogs incidentally caught

Visit of team BBMP to Veterinary college, Bangalore on 18th July 2019

ESTIMATION OF POST VACCINAL ANTIRABIES ANTIBODY TITER AND EVALUATION OF DIFFERENT REGIMEN OF VACCINATION IN CATTLE

Elimination of canine mediated human rabies by 2030 -Future strategies

• National Rabies Control Programme (NRCP) being revised by NCDC,

• Mass immunization - 70% dog population

• Awareness and education particularly : rural areas

• Immediate medical care after an animal bite.

• Uninterrupted availability of vaccines and RIG in all hospitals and in remote primary-health-care centres

• Pet ownership, Licensing of dogs, reducing the population of street dogs

• Availability of rabies diagnostic tests

Rabies elimination - an ideal mission for

‘One Health’ concept

NETWORK THROUGH POST GRADUATES IN FIELD….

2013 Molecular characterization: RT-PCR in animals

2014

2015

2016

2017

N gene –Molecular epidemiology and Development of LAMP

Status of antirabies vaccinal antibodies in dogs

Recombinant G protein for seromonitoring of vaccinal antibodies in dogs

Genetic Variability of G-L Intergenic region sequences of Indian RABV

Development of RFFIT for assessment of vaccinal efficacy in dogs

Comparative Evaluation Of LFA, LAMP with DFA and modified DFA

2018 Development of in-house ELISA for vaccinal antibodies

Workshop on Enhancing Progress Towards Rabies

Elimination ‘Zero by 30’ in the SAARC Region

26-28 June 2019, Kathmandu, Nepal

• Strengthening dog rabies surveillance

• Mass immunization of dogs

Achieving “ Canine mediated rabies elimination by 2030”

RINDERPEST

ONLY ONE SUCCESS STORY….

Next Rabies ????

Thank you all

KVAFSU ,CVA ,OIE, CDC, APHA and IVA

Lets look forward for Rabies free status!