SNAKES AND SNAKE VENOM

Gaurav kr singh
Research scholar
CHANDIGARH UNIVERSITY
 SNAKES ( OPHIDIA )
 

• There are more than 3500 species of snakes, but only about 250 are
venomous.
• In India about 216 species are found, of which 52 are poisonous.
• Only five are dangerously poisonous to man; king cobra, cobra, common
krait, Russell's viper and saw-scaled viper.
• The most common poisonous snake is common krait.
• The poisonous snakes may be divided into five families.
• (1)
• (A) VIPERIDAE - Russell's viper, gaboon viper, sawscaled viper, puff
adder.
• (B) Cortalidae - rattlesnakes, pigmy rattlesnakes, copper heads, cotton
mouths (water moccasins), pit viper, and the bushmaster.
• (2) Elapidae - Cobras, kraits, mambas, tiger snake, taipan, death adder,
copperhead snakes, coral snakes.
RUSSELS VIPER
Saw scaled viper
Rattle snake
Pit viper
Indian krait
Indian cobra
King cobra
Black mamba
Belchers or banded snake
(3) Hydrophidae - The sea snakes. All sea snakes are poisonous but they
seldom bite.

(4) Colubridae - Boom slangs, bird snake of the African continent

(5) Atractaspididae - African and Middle Eastern burrowing asps or stilleto

snakes also known as burrowing- or mole vipers or adders, or false vipers.
Mole viper
Boom slang
 Colubridae
• The colubridae or colubrine snakes lay eggs.
• Their head is of about the same width as that of the neck, and the pupils
of their eyes are circular.

 Elapidae
• The elapidae or land snakes have a round tail, and include the Cobra (naja
naja), the King Cobra or Hamadryad (Naja hanna), the Common krait
(Bengarus caerulus), the Banded Krait (Bangarus fasciatus), and the Viper
 Hydrophidae
• The Hydrophidae or sea snakes are found in the vicinity of the sea coast.
• Their eyes are very small and their tails are flattened like fins.
• Their nostrils are situated in the top of the snout and are valved so as to
enable them to breathe freely while swimming or in the sea.
• Their belly plates are not broad and the scales on their back are dull .
• Although poisonous, they are non-offensive by nature.
• However, their bite causes severe weakness due to muscle damage.
• The commonest species of the snake is Hydrophis caerulescens.
 Viperidae

• The Viperidae or viperine snakes have a peculiar broad, lozenge-shaped

(diamond) head, usually covered with small scales, a narrow neck and a
short tail.
• The pupils of their eyes are vertical slits.
• These snakes are divided into two main classes namely, pit vipers and
pitless vipers.
• Pit vipers have a pit or a deep depression on each side of the head
between the eye and the nostril, and usually occur in the hills. Their bite
is seldom fatal to man.
• Pitless vipers have no pit on the head.
• They have broad plates on the belly extending right across, and small
scales on the head similar to those on the body.

• The two species of pitless vipers which occur in India and are dangerous
to man are the Daboia or Russell's viper (Daboia or Vipera russellii or
Daboia elegans) and the Saw-Scaled Viper' Phoorsa or Echis (Echis
carinata),
• In India, the four to five medically important poisonous land snakes are—
the Indian Krait (Bengarus caerulus), the Common Cobra (Naja nag),
the Saw Scaled Viper (Echis carinatus) and the Russell's Viper (Viper
russelli).
 Cobra (Nag or Kal samp)
• Cobras are found throughout India.
• They a variable colour, although the colour is usually black.
• They are with a well-marked hood, which often bears a single spectacle
mark on the dorsal side, but sometimes, they have an oval spot
surrounded by an ellipse.
• The portion of the neck surrounding the spectacle mark is darker than the
rest of the back and is often speckled with small golden spots.
• Ordinarily, a cobra is seen without a hood. It expands its neck in the form
of a hood only when it is enraged, or is in danger or when it is about to
strike.
• The hood cannot be seen in a dead cobra, as the joints and the neck
become stiff.
Indian cobra
 King Cobra or Raj Nag or Raj Samp

• King cobras are mainly found in the Himalayas, lower Bengal, Assam
and Burma and in the hills and forests of southern India.

• They are bigger than common cobras, and grow to a length of eight to
twelve feet or even eighteen feet.

• They have a hood, which does not bear spectacle-marks.

• The young king cobra is jet black in colour, and is provided with white or
yellow cross bars on the body and tail, and four similar bars on the head.
COBRA vs KING KOBRA
• The adult king cobra varies a good deal in colour and may be yellow,
green, brown or black, and is usually provided with more or less distinct,
white or yellowish Cross bars or chevrons on the body.

• The belly may be nearly uniform, mottled or adorned with bars, while the
throat is usually light yellow or cream-coloured.
 Common Krait

• Common krait occurs throughout India, and is called maniyar in

Maharashtra, kalotaro in Gujarat, chitti in Bengal and kawriya or chit
kawriya in punjab.

• It varies from three to four feet or even five feet in length.

• It generally a shining steel-black colour, and has narrow single or double

white arches across the back.

• These arches begin at some distance from the head and extend up to
the tip of the tail.

• Its belly has a creamy white colour.

• A common krait can be identified by the following characteristic marks:
(i) the head is covered with large shields;

(ii) four shields are found on either side of the lower lip;

(iii) the scales in the central row down the back are large and hexagonal;

(iv) the tail is round;

(v) the plates under the tail like those on the belly are entire and not divided.
 Banded Krait

• The banded krait is larger than the common krait and grows to a length of
six feet, and rarely, even seven feet.

• It occurs in India in the north-east, and in far south in the basin of the river
Mahanadi.

• In addition to the distinguishing characteristic marks of the common krait,

the banded krait has alternate jet black and deep yellow bands across the
back.
  Russels viper

• Russels viper is also called Chain Viper, and is found throughout India
except the Gangetic valley.

• The species was named in after Patrick Russell (1726–1805), the

Scottish herpetologist that first described many of the Indian snakes.

• It is called kander in Hindi, ghonas in Marathi and chital in Gujarati.

• It has a light brown colour, and grows to a length of four to five-and-a-

half feet.
• It is stouter (FAT AND HEAVILY BUILT) than any other
poisonous snake in India, and narrows towards its tail, which is
short. head is flat, heavy and triangular, and has a white v-
shaped mark, with its apex pointing forward.

• The nostrils are bigger than those of any other Indian snake.
• The body is whitish with dark semi-lunar spots.

• It produces a terrible hissing sound when about to attack its

victim.

• The entire broad plates on the belly, the small scales on the
head and the shields beneath the tail divided into two rows are
sufficient to identify this snake
 Saw-Scaled Viper, Phoorsa or Echis (Echis carinata)

• The Saw-Scaled Viper is called phoodsa in Marathi and Gujarati, and

afai in Urdu.

• It is a small snake growing to a length of one-and-a-half feet, is brown

or brownish-grey in colour. It occurs in the states of Maharashtra,
Rajasthan, and other sandy parts of India.

• It has a triangular head, the upper surface of which is covered with a

white mark resembling a bird's footprint or an arrow.
• It has a continuous white wavy line along each flank of the back.
Diamond- shaped areas of a darker color are situated between the
upper curves of the two Wavy lines.

• The ridge in the middle of each scale is dented like a saw; hence it is
called a saw-scaled viper.
Differenciating between poisonous and non poisonous snakes
Snake bites and snake venom
• Snake venom is a complex mixture of organic compounds .

• Many of these compounds produce a variety of pathophysiological

effects including local tissue damage and/or systemic effects in the
affected individual.

• The major types of biomolecules found in snake venom are proteins,

some of which are enzymes whereas other proteins lack catalytic
activity.
 Mode of Ejection and Transmission of Venom

• In poisonous snakes, the poison is transmitted through the hollow teeth

known as fangs, which are the modified upper marginal teeth.

• These fangs are connected to the poison gland (venom gland) by

means of a duct.

• The venom gland in poisonous snake is a modified parotid salivary

gland, which secretes and stores the toxic saliva called venom. When
the snake bites, muscles compress the venom gland and force the
venom to come nut of the fangs.
TYPES OF SNAKE VENOM
 NEUROTOXINS
• Neurotoxins affect the nervous system.
A) An exchange of ions (charged atoms) across the nerve cell
membrane sends a depolarizing current towards the end of the nerve
cell (cell terminus).
B) When the depolarizing current arrives at the nerve cell terminus, the
neurotransmitter acetylcholine (ACh), which is held in vesicles, is
released into the space between the two nerves (synapse). It moves
across the synapse to the postsynaptic receptors.
C) ACh binds to the receptors and transfers the signal to the target
cell, after a short time it is destroyed by acetylcholinesterase.
Fasciculins:

• These toxins attack cholinergic neurons (those that use ACh as a

transmitter) by destroying acetylcholinesterase (AChE).
• ACh therefore cannot be broken down and stays in the receptor. This
causes tetany (involuntary muscle contraction), which can lead to
death.
• The toxins have been called fasciculins since after injection into mice,
they cause severe, generalized and long-lasting (5-7 h) fasciculations
(rapid muscle contractions).
• Snake example: found mostly in venom of mambas (Dendroaspis
spp.) and some rattlesnakes (Crotalus spp.)
Dendrotoxins:

• Dendrotoxins inhibit neurotransmissions by blocking the exchange of

positive and negative ions across the neuronal membrane lead to no
nerve impulse, thereby paralysing the nerves.

• Snake example: mambas

α-neurotoxins:
• Alpha-neurotoxins are a large group, with over 100 postsynaptic
neurotoxins having been identified and sequenced.
• α-neurotoxins attack the Nicotinic acetylcholine receptors of cholinergic
neurons.
• They mimic the shape of the acetylcholine molecule and therefore fit
into the receptors → they block the ACh flow → feeling of numbness
and paralysis.
• Snake examples: king cobra (Ophiophagus hannah) (known as
hannahtoxin containing α-neurotoxins), sea snakes (Hydrophiinae)
(known as erabutoxin), many-banded krait (Bungarus multicinctus)
(known as α-Bungarotoxin), and cobras (Naja spp.) (known as
cobratoxin)
CYTOTOXIC VENOM

• Cytotoxic venom is generally composed of several digestive enzymes

and spreading factors, which result in local and systemic injury.

• Clinically, local effects progressing from pain and edema to ecchymosis

(bleeds under the skin) and bullae (watery blisters) most commonly
predominate.

• Hematological abnormalities including benign defibrination with or

without thrombocytopenia (increased bleeding and decreased clotting)
may result, but severe generalized bleeding is not common.
• Pain and swelling occurs almost immediately after the bite from a
cytotoxic snake and gradually becomes worse, in the next few hours.
(Within 4 to 6 hours it will be more pronounced)
• It is often described as “cold fire” .Later shock develops and this may
cause death.
• Findings of necrosis usually are evident by 48 hours following the bite.
• Necrosis begins with darkening of the area around the fang punctures.
Blistering may follow. Necrosis usually is confined to the skin and
subcutaneous tissue, but may be quite extensive. A putrid smell is
characteristic.
• Acute inflammation of the eye follows venom-spitting exposure and is
characterized by ocular congestion, edema of the conjunctiva and
cornea, and a whitish discharge.
 CYTOTOXINS
Phospholipases:

• Phospholipase is an enzyme that transforms the phospholipid

molecule into a lysophospholipid (soap) ,the new molecule attracts and
binds fat and ruptures cell membranes.
• Snake example: Okinawan habu (Trimeresurus flavoviridis)
Cardiotoxins:

• Cardiotoxins are components that are specifically toxic to the heart. They
bind to particular sites on the surface of muscle cells and cause
depolarisation, the toxin prevents muscle contraction. These toxins may
cause the heart to beat irregularly or stop beating, causing death.

• Snake example: mambas, and some cobra species

Hemotoxins:
• Hemotoxins cause hemolysis, the destruction of red blood cells
(erythrocytes), or induce blood coagulation (clotting).

• Snake example: most vipers and many cobra species. The tropical
rattlesnake Crotalus durissus produces convulxin, a coagulant.
 Bungarotoxin
• Bungarotoxins are a group of closely related neurotoxic proteins
derived from the venom of kraits including Bungarus multicinctus.
• α-Bungarotoxin inhibits the binding of acetylcholine (ACh) to nicotinic
acetylcholine receptors; β- and γ-bungarotoxins act presynaptically
causing excessive acetylcholine release and subsequent depletion.
• There are four types:

1. α-Bungarotoxin
2. β-Bungarotoxin
3. γ-Bungarotoxin
4. k-Bungarotoxin
 CONSTITUENTS OF SNAKE VENOM
Enzymes

• Enzymes are biological (virtually always proteins) catalysts.

• A catalyst is a compound that increases the rate of the reaction but
does not affect the yield or equilibrium of the reaction.
• The enzymes in the snake venom can speed up chemical reaction
going on in an organism so much, that they can kill the organism.
• Enzymes bind temporarily to one or more of the reactants of the
reaction they catalyse.
• In doing so, they lower the amount of activation energy needed and
thus speed up the reaction.
The function of some of the key enzymes have been summarised in the
list below and described in greater detail down the page.

• Cholinesterase : attacks the nervous system, relaxing muscles to the

point where the victim has very little control.

• Amino acid oxidase : plays a part in digestion and the triggering of other
enzymes, (is responsible for venom's characteristic light yellowish
colouring.)

• Adenosine triphosphatase : believed to be one of the central agents

resulting in the shock of the victim and immobilizing smaller prey.
(probably present in most snakes.)
• Peptide bradykinin potentiators :- Greatly enhance one of the body’s
natural responses to injury (dilation and increased permeability of
blood vessels, stimulation of pain receptors, and contraction of some
smooth muscles), thereby enhancing diffusion of venom in the
bloodstream, increasing bleeding, and stopping the ability to flee.
Rattlesnakes contain this venom.

• Polypeptide toxins : Directly disrupt nerve-impulse transmission,

usually causing heart or respiratory failure. Snakes with this
compound in its venom: Mambas and colubrids : Cobra (Naja)
(cobratoxin), Cobra (Naja) (cardiotoxin), Rattlesnake ,Vipera
(viperptoxin).
• Proteolytic enzymes : Catalyse the breakdown of structural
components of tissues. All venomous species contain this venom.

• Hyluronidases : Catalyse reactions that break mucopolysaccharide

links in connective tissues, thereby enhancing diffusion of venom.
Several types of snake contain this venom.

• Proteases : Catalyse reactions that disrupt protein peptide bonds in

tissues, causing blood-vessel wall damage and haemorrhaging and
muscle-fibre deterioration. Vipers, pit vipers contain this venom.
• Phospholipases : Catalyses reactions that harm muscules and nerves.
Almost all venomous species (e. g. Copperheads, cottonmouths,,
Rattlesnakes, Cobras, Vipera)

• Thrombin like enzymes : Inhibit blood clotting. Vipers, pit vipers, a few
elapids (but rare) contain this venom.

• Nerve growth factor (an enzyme) : Stimulates the growth of nerve

cells. Copperheads and Cottonmouths, Rattlesnakes contain this
venom.
• Action : Disrupt normal cellular function, causing death of the affected
cells. Vipers and elapids (occurrences vary) contain this venom.
• Glycoproteins : Suppress normal immune response of tissues
through anticomplementary reactions. Some vipers contain this
venom.

• Biogenic amines : Disrupt normal transmission of nerve impulses

and other types of signalling between cells. Cottonmouths,
Copperheads, Rattlesnakes, Tree vipers, contain this venom.

• Other enzymes : ribonucleases, deoxyribonucleases, nucleotidases,

lactate dehydrogenases, acidic and basic phosphatases.
Local signs
• Fang marks
• Local pain
• Local bleeding
• Bruising
• Inflammation (swelling, redness, heat)
• Blistering
• Local infection,
• Necrosis
General symptoms
• Local effects:
— This is related to the digestive function of the venom and
causes local tissue necrosis.
— It is maximal with a viper bite and least with krait
• (so much so that the bite may go unnoticed and symptoms
which follow may not be attributed to snake bite).
• General:
— Nausea, vomiting, abdominal pain, weakness,
drowsiness,
• Cardiovascular (Viperidae):
— Visual disturbances, dizziness, faintness, collapse, shock,
hypotension, cardiac arrhythmias, pulmonary oedema.

• Bleeding and clotting disorders (Viperidae) — bleeding from recent wounds (including
fang marks) and from old partly-healed wounds —spontaneous systemic bleeding - from
gums, bleeding into the tears, rectal bleeding ,haematuria, vaginal bleeding, bleeding
into the skin (ecchymoses) ,intracranial hemorrhage..
• Neurological (Elapidae, Russell's viper):
Drowsiness, abnormalities of taste and smell, "heavy" eyelids, paralysis of facial muscles
and other muscles as well , difficulty in swallowing secretions, respiratory and
generalised flaccid paralysis .

• Skeletal muscle breakdown (sea snakes, Russell's viper): — Generalised pain, stiffness
and tenderness of muscles, trismus, myoglobinuria, cardiac arrest,
acute renal failure.
• Renal (Viperidae, sea snakes): Loin (lower back) pain, haematuria, haemoglobinuria,
myoglobinuria, symptoms and signs of uraemia (acidotic breathing, hiccups, nausea,
pleuritic chest pain)

• Endocrine (acute pituitary/adrenal insufficiency) (Russell 's viper)

— Acute phase: shock, hypoglycaemia
— Chronic phase (months to years after the bite): weakness, loss of secondary sexual
hair, testicular atrophy, hypothyroidism etc .
ANALYSIS OF SNAKE VENOM
• The analysis of snake venom is somehow different as routine
chemical tests are not applicable.

• The skin portion around the bite area may be a source of venom for
examination of its constituents by special tests viz. test for
cholinesterase or thromboplastin in venom, toxicity test, clot quality
test, preciptin test, gel- diffusion test and immuno-assays.
Toxicity Test:

• This test is a physiological test for confirmation of snakebite.

Procedure:
• The aqueous washings of the bite area or the serum of the victim is
injected into fowls with or without antivenin.
• The death of the fowl after injection of serum without antivenin and the
survival of the fowl after injection with antivenin after a few hours
confirms the snakebite.
 Precipitin Test for Snake Venom:

• The test depends upon the fact that when a foreign protein is injected
into an animal, certain specific antibodies known as preciptin are
formed in that animal's blood which have the capacity to precipitate
the specific foreign protein.

• The specific foreign protein, which causes the production of the

preciptin is known as preciptinogen or antigen.

Test Procedure:
• The aqueous wash or wash by isotonic solution of the bite area or the
serum of the victim is allowed to come in contact with antivenin (the
specific antivenin when the snake is identified or polyvalent antivenin
when the snake is not identified).
• A reaction between the two is then shown by the formation of a
cloudy precipitate at the line or plane of contact.

• This is sensitive for a very low dilution of protein (l in I ).

• This test can be done in small test tubes.

• The preciptin test may be done by new methods, which have

several advantages over the tube method viz. (l) very small quantity
of the extract and antivenin are required. (2) perfectiy clear extract
is not required,
 Gel Diffusion Test:
• The test is carried out on the thin layer of agar solution on a glass
plate or Petri dish.
• Small hole are punched out in the dried layer in a particular manner
i.e. One in the center and three or four round the circumference of a
circle of about 25 cm. in diameter.
• The wash from the bite area or serum of the victim (diluted with
isotonic saline solution) is placed in the central hole and the antivenin
in the other holes.
• Diffusion occurs if the reaction is positive and a line of precipitation
appear.
• This reaction is completed after an overnight diffusion.
 Immuno — Assays:
• These methods have recently been introduced. It is based on
antibody- antigen reaction which is very sensitive and specific.

• A labelled antigen or antibody is employed for the purpose.

• Other methods are based on the use of diverse traces in different

ways viz. an enzyme (in enzyme immuno-assay, EIA), or
erythrocyte (haemagglutination inhibition assay), a bacteriophage
(viroimmuno assay), a fluorescent group and a stable free radical
(FRAT, Spin immune-assay).
• Among the methods, enzymes are found to be most effective as these
exert catalytic action i.e. act as a amplifier and catalyze the formation of
more than 5 product molecules per minute.

• Labelled component of an antibody-antigen react on binds to its

complementary binding site and competes with unlabelled component.

• The concentration of the labelled component is measured by assaying

enzyme activity or radioactivity, which is proportional to the
concentration of antigen (protein) present in the sample.
• The commonly used types of enzyme immuno-assay are
homogeneous enzyme immuno assay or enzyme multiplied immuno-
assay (EMIT).

• Another type in which the antigen or antibody is linked to a solid-state

carrier is known as enzyme linked immunosorbent assay (ELISA).

• The above two techniques are simple, rapid and do not require pre-
treatment of biological sample. ELISA technique is expected to serve
the purpose of FSL for detection of snake venom in blood or washing
of the bite area.
Other Test:
• The aqueous extract (washings) of bite area may also be tested for
the presence of inorganic substance viz. sodium, potassium,
calcium and magnesium by spot tests and atomic absorption
spectrophotometer and also zincs, iron, cobalt, manganese and
nickel.
Samples to be collected in case of Snake Bite:
• The aqueous washing of bite area on a clear cotton swab (control
sample of cotton to be preserved also).

• Blood serum or blood of victim. The visceral tissues may be sent for
examination, if necessary to confirm the presence of other poison
(viscera is not suitable for detection of venom).