Janardan Saikia

Department of Entomology

Venoms
 Venoms are often characterized by median

lethal dose (LD50, LD50, or LD-50),
expressed in terms of mass fraction (e.g., mg
of toxin/kg of body mass)
 Kills 50% of victims of specified type
Poison is absorbed or ingested; a poisonous animal only
deliver toxic chemicals if another animal touches or eats it
 Venom, is always injected. Every venomous animal has
mechanism to inject toxins directly into another animal.

Different Source of Venom
Diversity
Vertebrates

Source
Spiders, Centipedes, Scorpions, Bees ,
Wasps and ants

Fish

Stingrays, sharks and chimaeras  etc.

Amphibians
Snakes

Salamandrid salamanders
Inland Tipan, Black mamba, Rattle snake,
Blue krait etc.

Other
reptiles

Mexican beaded lizard, gila monster and
minito lizard

Mammals

Solenodons, shrews and male platypus

Insect venom
Insect venoms vary significantly in their
composition.
Commonly contain complex mix of
proteins, peptides, and enzymes, as well as

C. 1988 . venom is of high importance in colony defense as primary function later as pheromone Venom glands in worker bees become active after adult emergence and maximal production achieved within two or three weeks after emergence Production is higher during summer months.Peak activity in the colony 2. due to 1.Relatively young individuals beginning their defense behavior HIDER R. Apidae family.Hymenopteran Order.

phospholipids etc. proteins. (The International Union of Immunological Societies) . 12% contains enzymes. amino acids. Venom is 88% water.Honey bee venom is odourless. clear and water-soluble. commercial preparations are brown Bee venom is produced by venom gland and Dufour's gland and stored in the venom sac. Dried venom is light yellow.

Composition of solid constituent of honey bee worker venom .

25 to 0.1316 pH Acid reaction Solubility Insoluble in alcohol Thermic effects Tolerates 100C Chemical stability Shows reducing activity against potassium carbonate. and hydrogen peroxide.0010 mg of dried apitoxin. . chlorine. potassium bichromate.35 mg of liquid apitoxin. that is 0. and vegetable papain and papayotin demises its activity Conventional units Bee sting brings in about 0.General properties of the Apis mellifera apitoxin Parameter Characteristic Specific gravity 1. Denaturated by ammonia. picric acid and potassium bichromate Enzymatic stability Pepsines. renine. pancreatine. bromine.

50μg of protein sample are subjected to each gel lane with three replications in each sample . Separation of honeybee (Apis mellifera) venoms manually extracted from venom gland (GV) and electrical stimulation (ESV) using one-dimensional gel electrophoresis.Fig.

2006]  Hyaluronidase facilitates diffusion of other venom constituents through the interstitial space [Ratcliffe et al.2011]  Api m 3.Melittin is major allergen responsible for intense local pain [Edstrom et. 2012] Phospholipase A-2 is important for specific IgE induction in sting victims [Putz et al. al. 5–12 are recognized as a strong IgE and T-cell response to bee venom of the sting victim .1992] and trigger the lysis of wide range of cells [Seppala et al.

Venom Synthesis Stage 1.17 mg venom/day .

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enables venom to as intracellular .Four levels of venom effects in humans Lev Active el substances 1 MCD What happens? Protease peptide (mast hyaluronidase cell destruction degranulating inhibitors protect from Hyaluronidase enzymatic breaks down peptides). reinforcement. hyaluronic acid into polymers that Protease function inhibitors.

Lev Active el substances 3 Apamin. se A and Histamine is released by Melittin action of MCD peptide . MCDpeptide. What happens? Apamin acts as a toxin on central nervous system Phospholipase A destroy and Phospholipa melittin RBC.

Mechanism Melittin a potent hemolytic that causes mast cell degranulation and activates PLA2 Which catalyzes the hydrolysis of phospholipid bonds. destroying major component of cell membranes and leading to cell death This cause an increase in synthesis of cytokines and stimulate arachidonic acid release resulting in an immune or .

Histamine causes The Natural Medicines Comprehensive dilation and increased Database. the adhesive properties of hyaluronic acid hold cells together. other venom components penetrate into cell Apamin blocks many inhibitory effects. by blocking calcium-dependent potassium channels MCD peptide are similar to apamin causing mast cell degranulation with release of histamine. 2009 .Hyaluronidase hydrolyzes hyaluronic acid.

2005.. Phys. J.helical peptide. C hat is Melittin ? Water soluble cationic amphipathic 26aa α.Qiu et al. Nonspecific cytolytic peptide that attack all lipids membranes ructure of Melittin  Powerful of itphospholipase In aqueous stimulator salt solution A2 exists as tetramer Bent alpha-helical rod Inner surface consistes of hydrophobic side chains Outter surface consists of hydrophillic side chains .

1990) Typical anti-m/o polypeptide Possess antibacterial and anti-inflammation function .Melittin Main component of bee venom 26 amino acid residues Used to study mechanism of translocation. incorporation of peptides and proteins into model biological membranes (Dempsey.

Case study Author s Hua et al. (2008) Work done  Melittin activities on Na+ -K+-ATPase and G-6-PD in membrane of RBC were studied Main mode of melittin inhibition to the activity of enzymes is the .

20 P1 producing rate/μmol L¯1 g¯1 h¯1 2. CInhibition of Ouabain.adhesion/insertion.11 0.08 1.69 A.12 0. D-Control .Table 1.06 0.78 Relative enzymatic activity/μmol L¯1 g¯1 h¯1 2.adhesion/insertion and free-state.27 2. Test result of the measurement of Pi producing speed and enzymatic activity Testing group A B C D A660nm 0. B.99 1.17 1.08 3.13 3.

Hua (2008) Chinese Journal of Biochemistry and b : Samples treated with melittin .The changing curve of melittin’s effect on Na + -K+-ATPase activity a : Relative enzyme activity RBC membrane samples treated with ouabain.

b’: Reaction initiated by NADP mixed . a’: Reaction initiated by NADP (without melittin).The velocity-time change curve II of G-6-PD catalysed NADPH production   Fig.

and thus Sonof et inflammatory al. Pharmacology & Thera reduces the generation mediators . which leads to its degradation and allows the NF-κB dimers to enter the nucleus. consists of kinases IKKα and IKKβ and regulatory subunit NEMO is a point of convergence for all 3 signaling pathways  The IKK complexes phosphorylate IκBα. 2007.. where they bind to the cognate DNA binding sites and activate the expression of the pro-inflammatory gene  BV (melittin) inhibits the release of IκB through the inhibition of IKKs  Which results in NF-κB inactivation. NF-κB activity is stimulated by stimuli  The IKK complex.

phosphorus. IκB. NF-κB. IKK. inhibitor of NF-κB. . Anti-arthritic effect of BV (melittin). P. nuclear factor-κB.Fig.

Effects of melittin to dissociated rat dorsal root ganglion neurons were studied  whole-cell patch clamp and  calcium imaging techniques  Melittin induced intracellular calcium increases in 60% of small (<25 µm) and medium (<40 µm) diameter sensory neurons. Most melittin-sensitive neurons were .

Melittin (Mel)-induced rise in intracellular Ca2+ concentration and action potential firing in DRG neurons .Fig.

2010  Bee venom serine protease (Bi-VSP) is a multifunctional enzyme  In insects. the triggering phenoloxidase factor (PO) cascade Bi-VSP when injected induces a melanization response in target insects  lethal . it acts as arthropod prophenoloxidase (proPO)-activating (PPAF).How can venom act on insects ? Author Work done s Choo et al.

(A) PO activity in hemolymph of fifth-instar  B. .Fig. mori larvae injected with Bi-VSP (5 µg/larva). mori larvae injected with Bi-proVSP (5 µg/larva). (B) Immunofluorescent staining of hemocytes and fat bodies in fifth-instar B. Bi-proVSP (5 µg/larva).

2001 .Phospholipase A2 Inflammation Signaling Pathway Marr and Acorn.

obesity. dependent on the subsequent enzyme or decrease the transcription rate of stearoyl-CoA. It is largely . Protein Kinase C then initiates a signaling cascade that results in the activation of (mitogen-activated protein kinase) MAPK MAPK phosphorylates PLA2. desaturase 1 and glucose transporter 4 These endpoints have been linked to asthma. rheumatoid arthritis. and this active phospholipase cleaves intracellular membrane lipids to form a lysolipid and a fatty acid The fatty acid is often arachidonic acid (disease pathway) Arachadonic acid can enter three different pathways. and it activates protein kinase C. atherosclerosis.protein signalling cascade and is linked to phospholipase C Diacylglycerol is a cleavage product of phospholipase C. and diabetes.

and beeswax Bee venom therapy is the use of live bee stings (or injectable venom) to treat various . royal jelly.Use of honey bee products for medical purposes. raw honey. pollen. propolis. this include bee venom.

APITHERAPY SOCIETIES WORLDWIDE International Federation of Apitherapy American Apitherapy Society  Italian Apitherapy Association Malaysian Apitherapy Society …etc.  .

How Apitherapy is Administered Healthcare providers and apitherapists Is patient allergic to venom? No allergic reaction the therapy is continued Carried out every other day Increasing number of bee stings or injections Length of treatment determined by condition that is being treated .

Beneficial effects of whole bee venom in animal and cells Sl. bladder and renal cancers cells by different mechanisms of action depending on the tumor type.Table.no Effect or Target Specific effects 1 Anti-inflammatory and anti-arthritis action Glucocorticoid-and aspirin like effects 2 Anti-cancer effects Antitumor effects on ovary. PNS) • Stimulates many peripheral chemoreceptors. prostate. 3 Affects the central and peripheral nervous system (CNS. hepatoma. affecting flow to the CNS • Has cholinolytic action (against acetylcholine) • Blocks transmission of the vegetative synapse and the polysynaptic neuronal paths • Pain-soothing aspirin-like action • Management of chronic and inflammation pain • Influence of brain EEG and behaviour patterns • Increases brain blood circulation .

no Effect or Target Specific effects 4 Heart and system blood •Increases coronary and peripheral blood circulation. Leukaemia and HIV viruses. and inactivation of Herpes.bee- . 8 Endocrinological system Increases secretion of thyroid. hypophysis and of the hypothalamus hormones 9 Wound healing Promotes skin cell regeneration 10 Anti-diabetic Lowers blood glucose and increases insulin secretion Source: Bee Product Science. www. • Slows down heart at lower doses •Against blood coagulation fibrinolytic.Sl. stimulates the building of erythrocytes 5 Action on the immune system Immunosuppressive and immunoactivating 6 Protection from radiation Improves regeneration of leucocytes and erythrocytes 7 Antibiotic fungicide and antiviral action Bactericide action against different pathogens Action against Candida albicans.

05 mg/kg/day) failed to show anti-inflammatory or antinociceptive effects on RA Young.9 mg/kg/day) resulted in new bone proliferation and soft tissue swelling  While BVE treatment (0. al. et.Anti-arthritic actions of bee venom (BV) acupuncture two fractions Water soluble fraction (BVA) and  Ethylacetate soluble fraction (BVE)  BVA injection (0.2012.. Life Scienc  .

Fig. (C) water soluble fraction of BV (BVA) treated arthritic animals and (D) ethylacetate fraction of BV (BVE) . (A) The X-ray images at 3 weeks after arthritis induction in normal animals. (B) saline-treated arthritic animals.

 Yong. BVA (0. 3 weeks after first immunization. but not by opioid receptor antagonist.25 mg/kg) injected resulted in antinociceptive effect  Antinociceptive effect of BVA was blocked by α2-adrenergic receptor antagonist  2 mg/kg. 2mg/kg..al. pretreatment. et. studied antinociceptive effect with opioid and α2-adrenergic mechanism of BVA in the collagen-induced arthritis (CIA) rat model  Results showed.(2006). pretreatment .

Fig. . Changes of tail flick latency (TFL) after induction of collagen-induced arthritis (CIA).

diminishes surface tension of membranes Anti-inflammatory. prevents neuronal cell death *Induces inflammation.n o 1 Component % Melittin (Biologically active peptide 50-55 %) Effects • • • Membrane-active. Stimulates smooth muscles Increases capillary permeability increasing blood circulation and lowering blood pressure *Higher doses haemolytic 2 Phospholipase A (Enzyme hydrolysing phospholipids 10-12 %) • • are inflammatory and Destroys phospholipids and dissolves cell membrane of blood bodies Lowers blood coagulation and blood pressure. strongest thus most harmful BV Component allergen.Main biological and therapeutic effects of bee venom and its components Sl . .

antiserotonine action • Increases defense capability. • Lyses mast cells. Immuno-supressor. thus enabling penetrating of BV 5 Apamine (Biologically active peptide 2-3 %) • Anti-inflammatory stimulating release of cortisone. releasing histamine. serotonine (mast cell and heparine Degranulatingpepti • Increasing capillary permeability de 2-3 %) • Anti-inflammatory and simulates CNS • Dilates blood vessels and increases permeability *Allergenic . the tissue cement 1-2 %) • Catalyses the hydrolysis of proteins. stimulates CNS *Higher doses are neurotoxic 6 MCD.Sl .n o Component % Effects 3 Phospholipase B (cleavage of the toxic lysolecetin 1%) Detoxicating activity 4 Hyaluronidase (Catalyses hydrolysis of hyoloronic acid.

net. plasmin. thrombin Decreasing inflammation.Sl Component % . www.bee-hexagon. chymotprypsin. *Allergenic (Neurotransmitte r 0. Procamine 3-5 % Antiradiation effects 10 Histamine Dilates blood vessels. tertiapin.5 %) Source: Bee Product Science.7-1. cardiopep. decreases pain Inhibits aggregation of erythrocytes 8 ProteaseInhibitors (Biologically active Peptides 3-4 %) Inhibits activity of different proteases like trypsin. anti-rheumatic 9 Secapin.Feb’20 .n o Effects 7 Adolapin (Biologically active Peptide 1 %) Inhibits specific brain enzymes cyclooxigenase and lipooxigenase Decreases inflammations by anti-rheumatic.

Commercial Products .

royal jelly. rheumatoid arthritis. Bee venom is used to manufacture lotions. this include bee venom. low back pain etc. multiple sclerosis. pollen. ointments and injected preparations The bee venom is safe for human treatments. balms. and beeswax Bee venom therapy is the use of live bee stings (or injectable venom) to treat various diseases such as arthritis. the . raw honey.Conclusion Apitherapy is the use of honey bee products for medical purposes. propolis.

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