1. SNAKE BITE STUDY - PROTOCOL DEVELOPMENT WORKSHOP March, Organized by Christian Medical College, Vellore
Presentation by
Dr. A. K. Mukherjee
Professor, Department of Molecular Biology and Biotechnology,
Tezpur University, Tezpur, Assam
Head and Coordinator, DBT Nodal Center for Medical Colleges and Biomedical Research Institutes of NE India, T.U.
Affiliate Professor, School of Biological Sciences,
University of Northern Colorado, Greeley, USA

2. Strategies to Improve Snakebite Treatment: Challenges and Progress

3. Efficient Management of Snakebite in India: Problems Encountered
The following constrains as listed below are based on my 18 years of research experience on snake venom and snakebite management in India:
No proper /methodical (computerized) record of snakebite data in most of the Govt. hospitals: lack of region specific true snakebite data.
Geographical variation in snake venom composition and a detail study on region specific clinical manifestations following envenomation is lacking
Lack of reliable venom detection kit : failure to understand which species of snake is bitten?
Snakebite treatment protocol-which guideline is to be followed?
Quality control (efficiency) and supply of antivenom: Who have been taking care?
Monovalent Vs Polyvalent antivenom in snakebite therapy: Superiority of treatment Vs treatment cost needs to be worked out.
Less attention has been paid for very poorly immunogenic, low molecular mass venom components which are difficult to neutralize by antivenom.
Venom quality control for efficient antivenom production: should be addressed

4. Antivenom manufacturers
All these problems have aroused because the lack of a proper coordination amongst……
Clinicians
Toxinologists
Antivenom manufacturers
To address all these issues, “Toxinological Society of India” was established in 2011

5. Hospital based clinical work
For Efficient Management of Snakebite, the Following Scheme is Proposed
Snakebite Management
Laboratory based venom and antivenom analysis
Hospital based clinical work
Improvement of treatment & quality of antivenom

6. Laboratory Based Snakebite Management
Venomics
Antivenomics
Venom detection kit
A proteomics approach to study geographical variation in venom composition & pathophysiology of venom components
Proteomics based immunochemical analysis of antivenom
ELISA based affordable venom detection kits
Molecular basis of taxanomic identification of Indian snakes

7. Venomics: A proteomics Approach for Analysis of Snake Venom Components
Analysis of complex mixture of snake venom demands an approach that effectively separates a complex mixture of proteins and peptides with varying size and charge.
Traditional methods of venom analysis does not provide understanding of the complexity of venom toxicity as well as elucidating the major biochemical differences among snake families.
Efficiency of the current proteomic techniques in identifying pharmaceutically important venom proteins and a better correlation of geographical variation in venom composition with clinical manifestation in snakebite patients has been proved
Advantages
characterized snake venom proteomes from different parts of India would provide information useful with regard to the potential clinical manifestation of these bites in a particular location
This will also guiding antivenom choices for proper treatment of snakebite victims.

8. Differences in the venom composition between two species of Indian
Cobra (Naja naja and Naja kaouthia)
Impact on the severity of
pathogenesis and antivenom treatment

9. Pictures of two specis of Indian Cobra
Naja naja
Naja kaouthia

10. Although the biochemical composition of Naja naja from different parts of India is well Characterized, but no information was available on the biochemicalcomposition and biological activity of N. kaouthia venom from India.
Therefore, effort was given to characterize the biochemical properties of N. kaouthia venom from eastern India and to compare these properties with the N. naja venom of the same origin.
Further, potency of a commercial polyvalent antivenom raised against N. naja in neutralizing the lethality and various pathophysiological effects of N. kaouthiaVenom was also explored.

11. A comparison of enzyme activity present in
N. naja and N. kaouthia
venom samples from
eastern India

12. Source: Mukherjee, A. K. and Maity, C. R
Source: Mukherjee, A.K. and Maity, C.R. (2002) Comparative Biochemistry and Physiology 131 (B),

13. Comparison of gel-filtration pattern of
N. naja and N. kaouthia venom samples
from eastern India

14. Naja Naja
venom
Naja Kaouthia
venom

15. Properties Naja naja Naja kaouthia
Table: A Comparison of Pathophysiology between N. naja and N. kaouthia
Properties
Naja naja
Naja kaouthia
Lethality (mg/body weight of mouse)
0.40 ± 0.08
0.7 ± 0.09
Hemorrhagic activity (20μg crude venom)
Nil
Myotoxicity (CPK release) (1U/l)
28.5 ± 3
19.2 ± 2.1
Edema ratio (15μg crude venom)
148.0 ± 3.1
140.0 ± 3.1
Minimum edema inducing dose (μg)
5.0
4.8
Indirect hemolytic activity (% of Hb release by 100 μg of protein)
86 ± 4.1
80 ± 3.8
Direct hemolytic activity (% of Hb release by 100 μg of protein)
50 ± 2.1
39 ± 2.5
Neurotoxicity
Present
Source: Mukherjee, A.K. and Maity, C.R. (2002) Comparative Biochemistry and
Physiology 131 (B),

16. Neutralization of lethality and pathophysiology of N. naja and
N. Kaouthia Venom samples by a commercial N. naja antivenom
(mg venom / 100 mg antivenom)
Neutralization potency

17. Antivenomics: Application of Proteomic Tools for Production of Efficient Antivenom for Proper Management of Snakebite Patients
Proteomics-based immunochemical analysis (antivenomics) provides relevant information for outlining which venom mixtures cross-react with the most important components in medically-relevant venoms from a particular region.
This type of approach may set the basis for the development of effective antivenoms on an immunologically sound basis which in turn will be beneficial for better treatment of snakebite patients

18. Venom Detection Kits
Background Information:
The diagnosis of snake envenomation is based on sensitive immunoassays
Geographical and species specific variation in snake venom composition as well as high-immunological cross-reactivity impose a great challenge for development of ELISA-based venom detection kits.
Recently, Wahby and Ibrahim (2008) and ibrahim et al. (2013) have demonstrated avidity based VDK
(Avidity: an inexact measure of the binding strength of antibodies to multiple antigenic determinants on natural antigens)

19. Identification and Discrimination of Snake Venoms from Egyptian Cobras
Source: Ibrahim et al. Toxicon 63 (2013) 88-97

20. Permission to collect snake venom from different geographical location
IMPROVEMENT OF SNAKEBITE TREATMENT: SUMMARY OF SUGGESTIONS
To identify PI’s from different fields but having a similar research interest.
To identify coordinator(s) from each group, may be one from each region of India
Snakebite data collection from different parts of country-permission from Govt.
Permission to collect snake venom from different geographical location
Approach to Govt. of India for allocation of more money in snakebite research
Organization of workshop for project writing
Organization of regular meetings amongst the interested PIs.
Strengthening of Toxinological Society of India (TSI) and its activities