SNAKE BITE STUDY - PROTOCOL DEVELOPMENT WORKSHOP
11-12 March, 2013
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
Strategies to Improve
Snakebite Treatment:
Challenges and Progress
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:
1. No proper /methodical (computerized) record of snakebite data in most of the Govt.
hospitals: lack of region specific true snakebite data.
2. Geographical variation in snake venom composition and a detail study on region specific
clinical manifestations following envenomation is lacking
3. Lack of reliable venom detection kit : failure to understand which species of snake is bitten?
4. Snakebite treatment protocol-which guideline is to be followed?
5. Quality control (efficiency) and supply of antivenom: Who have been taking care?
6. Monovalent Vs Polyvalent antivenom in snakebite therapy: Superiority of treatment Vs
treatment cost needs to be worked out.
7. Less attention has been paid for very poorly immunogenic, low molecular mass venom
components which are difficult to neutralize by antivenom.
8. Venom quality control for efficient antivenom production: should be addressed
All these problems have aroused because the lack of a proper coordination
amongst……
Clinicians
Antivenom
manufacturers
Toxinologists
To address all these issues, “Toxinological Society of India” was established in 2011
For Efficient Management of Snakebite, the Following Scheme is Proposed
Snakebite
Management
Hospital based
clinical work
Laboratory based
venom and
antivenom analysis
Improvement of
treatment & quality
of antivenom
Laboratory Based Snakebite Management
Venomics
A proteomics
approach to study
geographical
variation in venom
composition &
pathophysiology of
venom components
Antivenomics
Proteomics based
immunochemical
analysis of
antivenom
Molecular basis of taxanomic
identification of Indian snakes
Venom
detection kit
ELISA based
affordable venom
detection kits
Venomics: A proteomics Approach for Analysis of Snake Venom Components
1. 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.
2. 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.
3. 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.
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
Naja naja
Naja kaouthia
• 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.
Source: Mukherjee, A.K. and Maity, C.R. (2002) Comparative Biochemistry and Physiology 131
(B), 125-132
Naja Naja
venom
Naja
Kaouthia
venom
Table: A Comparison of Pathophysiology between N. naja and N. kaouthia
Properties
Lethality (mg/body weight of mouse)
Naja naja
0.40 ± 0.08
Hemorrhagic activity (20μg crude venom) Nil
Naja
kaouthia
0.7 ± 0.09
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 50 ± 2.1
by 100 μg of protein)
39 ± 2.5
Neurotoxicity
Present
Present
Source: Mukherjee, A.K. and Maity, C.R. (2002) Comparative Biochemistry and
Physiology 131 (B), 125-132
Neutralization of lethality and pathophysiology of N. naja and
N. Kaouthia Venom samples by a commercial N. naja antivenom
0.9
Neutralization potency
(mg venom / 100 mg antivenom)
0.8
0.7
0.6
0.5
N. naja
N. kaouthia
0.4
0.3
0.2
0.1
0
Lethality
Edema
inducing
Direct
hemolysis
indirect
hemolysis
Myotoxicity
Antivenomics: Application of Proteomic Tools for Production of Efficient
Antivenom for Proper Management of Snakebite Patients
1. 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.
2. 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
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)
Identification and Discrimination of Snake Venoms from Egyptian Cobras
Source: Ibrahim et al. Toxicon 63 (2013) 88-97
IMPROVEMENT OF SNAKEBITE TREATMENT: SUMMARY OF SUGGESTIONS
1. To identify PI’s from different fields but having a similar research interest.
2. To identify coordinator(s) from each group, may be one from each region of India
3. Snakebite data collection from different parts of country-permission from Govt.
4. Permission to collect snake venom from different geographical location
5. Approach to Govt. of India for allocation of more money in snakebite research
6. Organization of workshop for project writing
7. Organization of regular meetings amongst the interested PIs.
8. Strengthening of Toxinological Society of India (TSI) and its activities