CHAPTER 7
VENOM

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7.1 Introduction

Among the many species of insects, only very few have the capability of defending themselves with a sting and venom injection during stinging. All insects that can sting are members of the order Hymenoptera, which includes ants, wasps and bees. Since the sting is believed to have evolved from the egg-laying apparatus of the ancestral, hymenopteran species, only females can sting. The sting is always at or near the abdominal end, rather than the head. Therefore the pain inflicted by a honeybee, defending its colony, is not caused by a bite, as is frequently said, but by a sting.

There are many other poisonous insects which secrete venom. They usually cover their body with it, spray it, create wounds and secrete it into the wound, or inject it via mouthparts or a sting. In some cases, the venom is used for defense of the individual or, in the case of social insects, the colony. But venom is also used in killing prey (as with some wasps or spiders) or for immobilizing and preserving prey (for their own or their developing offspring's consumption).

a)
Figure a) unique to the honeybee sting. Once stung by a honeybee, the whole sting apparatus, venom sack and all, almost always remains
b)
Figure b) This occurs only with honeybees and with no other stinging insect.

Figure 7.1 : A honeybee worker, stinging the relatively tough human skin, is unable to withdraw its sting lancets because of the fine barbs (a) unique to the honeybee sting. Once stung by a honeybee, the whole sting apparatus, venom sack and all, almost always remains (b). This occurs only with honeybees and with no other stinging insect.

Honeybee venom is produced by two glands associated with the sting apparatus of worker bees. Its production increases during the first two weeks of the adult worker's life and reaches a maximum when the worker bee becomes involved in hive defence and foraging. It diminishes as the bee gets older. The queen bee's production of venom is highest on emergence, probably because it must be prepared for immediate battles with other queens.

When a bee stings, it does not normally inject all of the 0.15 to 0.3 mg of venom held in a full venom sac (Schumacher et al., 1989 and Crane 1990, respectively). Only when it stings an animal with skin as tough as ours will it lose its sting - and with it the whole sting apparatus, including the venom sac, muscles and the nerve centre (see Figure 7.1 and 7.2). These nerves and muscles however keep injecting venom for a while, or until the venom sac is empty. The loss of such a considerable portion of its body is almost always fatal to the bee.

 

If disturbed or handled improperly most colonies will defend themselves.  Honeybees in many parts of the world are very sensitive to disturbances and react en masseto defend their nest, as this innocent dog found out on approaching beehives recently inspected by beekeepers in northern Argentina.  With some help from an emergency sting kit (epinephrine injection and antihistamine tablets) the dog survived more than 1000 bee stings.

Figure 7.2 : If disturbed or handled improperly most colonies will defend themselves. Honeybees in many parts of the world are very sensitive to disturbances and react en masse to defend their nest, as this innocent dog found out on approaching beehives recently inspected by beekeepers in northern Argentina. With some help from an emergency sting kit (epinephrine injection and antihistamine tablets) the dog survived more than 1000 bee stings.

The median lethal dose (LD50) for an adult human is 2.8 mg of venom per kg of body weight, i.e. a person weighing 60 kg has a 50% chance of surviving injections totalling 168 mg of bee venom (Schumacher et al., 1989). Assuming each bee injects all its venom and no stings are quickly removed at a maximum of 0.3 mg venom per sting, 600 stings could well be lethal for such a person. For a child weighing 10 kg, as little as 90 stings could be fatal. Therefore, quick removal of the stings is important. However, most human deaths result from one or few bee stings due to allergic reactions, heart failure or suffocation from swelling around the neck or the mouth.

Used in small doses however, bee venom can be of benefit in trcating a large number of ailments. This therapeutic value was already known to many ancient civilizations. Today, the only uses of bee venom are in human and veterinary medicine.

7.2 Physical characteristics of venom

Honeybee venom is a clear, odourless, watery liquid. When coming into contact with mucous membranes or eyes, it causes considerable burning and irritation. Dried venom takes on a light yellow colour and some commercial preparations are brown, thought to be due to oxidation of some of the venom proteins. Venom contains a number of very volatile compounds which are easily lost during collection.

7.3 The composition of venom

A large number of studies have been carried out on the composition of honeybee venom. Much of the basic identification of compounds, their isolation and the study of their pharmacological effects was done in the 1950's and 1960's. There are some comprehensive summaries in Piek (1986) which cover the morphology of the venom apparatus, the collection of venom, the pharmacological effects of bee venom and allergies to the Hymenoptera venom of bees, wasps and ants.

88% of venom is water. The glucose, fructose and phospholipid contents of venom are similar to those in bee's blood (Crane, 1990). At least 18 pharmacologically active components have been described, including various enzymes, peptides and amines. Table 7.1 lists the major components as summarized from Dotimas and Hider (1987) and Shipolini (1984). No further discussion of the detailed chemistry and various effects of individual components will be attempted here. Schmidt (1992) presents a comprehensive account of allergies to honeybee and other Hymenoptera venoms. Crane (1990), Dotimas and Hider (1987) and Banks and Shipolini (1986) give a very good overview of its composition, effects, harvesting and use.

Venom from other Apis species is similar, but even the venoms from the various races within each species are slightly different from each other. The toxicity of Apis cerana venom has been reported to be twice as high as that of A. mellifera (Benton and Morse, 1968).

Table 7.1:
Composition of venom from honeybee worker

Class of molecules

Component

% of dry venoma

% of dry venomb

Enzymes Phospholipase A2

Hyaluronidase

Acid Phosphomonoesterase

Lysophospholipase

a -glucosidase

10-12

1-3

10-12

1.5-2.0

1.0

1.0

0.6

Other proteins and peptides Melittin

Pamine

Mast Cell Degranulating Peptide (MCD)

Secapin

Procamine

Adolapin

Protease inhibitor

Tertiapinc

Small peptides (with less than 5 amino acids)

50

1-3

1-2

0.5-2.0

1-2

 

0.1

13-15

40-50

3

2

0.5

1.4

1.0

0.8

0.1

Physiologically active amines Histamine

Dopamine

Noradrenaline

0.5-2.0

0.2-1.0

0.1-0.5

0.5-1.6

0.13-1.0

0.1-0.7

Amino Acids t -aminobutyric acid

a -amino acids

0.5

1

0.4

Sugars Glucose & fructose

2

 
Phospholipids  

5

 
Volatile compounds  

4-8

 

Dotimas and Hider, 1987; b Shipolini, 1984
This peptide may not be present in all venom samples

7.4 The physiological effects of venom

7.4.1 Unconfirmed circumstantial evidence

Bee venom has long been used in traditional medicine for the treatment of various kinds of rheumatism. Although venoms of the different honeybee species differ slightly, there have been reports of successful rheumatism treatment with Apis dorsata venom by Sharma and Singh (1983) and with A. cerana venom by Krell (1992, unpublished).

The list of benefits to human beings as well as to animals is very long. Most of the reports of cures are of individual cases, though several unrelated patients have experienced the improvement or cure of similar ailments. Bee venom treatments are often accompanied by changes in life style, nutrition or other which may account for part, if not most of the benefits from treatments. Reported clinical tests were often conducted in countries with less rigorous methods than the standard Western, double-blind placebo tests. Despite these considerations, many patients did report positive results and many of the successful treatments occurred after established medical or surgical procedures had failed. However, there is a very real resistance in Western medical circles either to accept these results or to test bee venom treatments according to Western medical standards.

The diseases and problems which have been reported by patients or doctors as improved or healed with bee venom therapy are listed below (Table 7.2). This does not constituent an endorsement or recommendation for the treatments. Stinging should never be tried unless there is immediate access to emergency treatment in case of an allergic reaction.

Table 7.2 List of diseases and health problems improved or healed according to anecdotal reports

Humans
Arthritis, many typesa

Epilepsya

Mastisa

Chronic paina

Decreases blood viscosity and coagulabilityb

Neruosesb

Therosclerosisd

Infectious spondylitise

Infect. Polyarthritise

Myositisf

Thrombophiletritisf

Iritisg

Animals

Arthritis

Multiple sclerosisa

Bursitisa

Some types of cancera

Migraineb

Dilates capillaries and arteriesb

Rhinosinusitisc

Polyneuritise

Neuralgiae

Malariae

Tropical ulcersf

Cancer, temporaryf

Iridocytisg

Premenstrual syndromea

Ligament injuriesa

Sore throata

General immuno-stimulant

Decreases blood cholesterol levelb

Endoarteriosisd

Radicultitisef

Endoarthritise

Intercostal myalgiaf

Slowly healing woundsf

Keratoconjunctivitisg

Asthmah

BeeWell, 1993, 1992; b Kel’man, 1960; c Fotin & Gel’medova, 1981; d Poryardin, 1960;
e Gaider, 1950; f Lavochev, et al., 1958; g Naum Iyorish, 1974; h Dutta, 1959
.

7.4.2 Scientific evidence

During the last seven decades, over 1700 scientific publications on the composition and various effects of bee venom in animals and humans have been published. An overwhelming proportion comes from Eastern Europe and Asia. Most of them concentrate on demonstrating the site specific, physiological effects of individual components such as membrane destruction, toxicity, or the stimulation or blocking of enzyme reactions. This has largely increased our understanding of the processes occurring after a sting, the physiological effects of isolated venom compounds and the substances responsible for most of the allergic reactions. It has contributed little to verifying the increasing claims of different therapeutic values attributed to honeybee venom, however.

A study with whole bee venom on dogs (Vick and Brooks, 1972) and rats (Dunn, 1984) showed that melittin and apamine produce increased plasma cortisol. Together with various other arguments, this suggests that many of the curative effects of bee venom may work through stimulation of the body's enzyme and immune system, in a way similar to the common drug cortisone. Cortisone has been used in the treatment of many ailments, but it is also known to have strong, undesirable side-effects. Melittin also appears to have toxic side effects as do some of the other individual compounds in venom. When whole venom is applied however, no side-effects have been shown, other than in allergic patients (Broadman, 1962 and Weeks, 1992 personal communication).

The anti-inflammatory effects of bee venom are perhaps the best studied and the various mechanisms have been repeatedly described in scientific literature (Rekkaand Kourounakis, 1990; Kim, 1989 and others). The neurotoxic venom compounds have shown a potential benefit for epileptic patients (Ziai, 1990). The protective value of bee venom and melittin against the lethal or damaging effects of x-rays has been investigated (Shipman and Cole, 1967 and Ginsberg et al., 1968). Though these and many other results are encouraging, no clinical studies have been carried out to verify the effectiveness using tests accepted by the Western medical establishment. Nevertheless, more and more physicians and healers are experimenting with this benign treatment after they have tested the patient's allergic reactions to bee venom. Recently, after long efforts by the American Apitherapy Society and its members, some interest has been shown by national institutions in several Western European countries and the USA for clinical and large scale tests of bee venom therapy.

A good summary of the scientific studies, with further references can be found in Banks and Shipolini (1986) and Schmidt (1992). Summaries of some of the major specific effects of the various venom compounds that are shorter and more easily understood, can be found in Mraz (1983), Dotimas and Hider (1987), Crane (1990) and Schmidt and Buchmann (1992). The American Apitherapy Society keeps records of scientific as well as anecdotal information on the use of bee venom. It is also probably the best source of information on any subject related to apitherapy (see Annex 2).

7.5 The use of venom today

No uses for venom, other than medical ones are known to the author. The only legally accepted medical use of bee venom in Western European and North American countries is for desensitizing people who are hypersensitive (allergic) to bee venom. Since the early 1980's, pure bee venom has been used for desensitization. The use of whole body extracts has been largely discontinued after a double-blind test proved the higher efficiency of pure venom (Hunt et al., 1978). In Eastern Europe and in many Asian countries bee venom has been used in official medical treatment of a large variety of ailments for a considerable length of time.

The use of pure venom injections and well placed bee stings is increasing in Western countries as an alternative to heavy (and soinetimes ineffective) drug use, which is often associated with numerous side-effects. This is particularly so for arthritis and other rheumatoid inflammations. A list of some other ailments for which successful treatments with bee stings have been reported has been given in section 7.4.1.

Application methods for venom include natural bee stings, subcutaneous injections, electrophoresis, ointments, inhalations and tablets (Sharma and Singh, 1983).

Since bee venom has both a local and a systemic effect, correct placement of injections, or stings and the dosage are very important. Therefore, bee venom therapy must be properly learned. Still, relief of some ailments can be obtained by simply applying a sting or two to the affected area, i.e. to some painful, immobile arthritic joints.

A society for api-acupuncture was formed in 1980 in Japan (see Annex 2). In the following years, many reports of experiences and successes in api-acupuncture appeared (in Japanese) in Honeybee Science (e.g. Ohta, 1983 and Sagawa, 1983). In the Republic of China, bee venom therapy is combined with a knowledge of acupuncture by many hospitals and physicians.

In the West, the American Apitherapy Society (AAS) is collecting case histories and information on bee venom therapy, together with medical uses of other bee products. There may be other national organizations, particularly in Eastern Europe and Asia. IBRA and Apimondia also have a wide collection of reference materials (see Annex 2).

7.6 Venom collection

Early collection methods required surgical removal of the venom gland or squeezing each individual bee until a droplet could be collected from the tip of the sting. Since the early 1960's, extraction by the electro-shock method has been continuously improved and is now standard procedure.

Different extraction or collection methods result in different compositions of the final products Venom collected under water to avoid evaporation of very volatile compounds seems to yield the most potent venom (Pence, 1981). Venom collected from surgically removed venom sacs showed different protein contents from that collected with the electroshock method (Hsiang and Elliott, 1975). Gunnison (1966) used a cooling system with the standard electro-shock collecting apparatus in order to preserve more of the volatile compounds.

The standard electro-shock method (Morse and Benton, 1964a, b) cannot be recommended for venom collection from Africanized honeybees or the more defensive races in other parts of the world. Colony arousal can become so overwhelming that bees start killing each other and alert other colonies or attack the beekeeper and bystanders. The mass reaction of Africanized honeybees may also result in contamination of the collected venom. Nevertheless, venom is collected by this method in Brazil and Argentina, with only minor modifications.

Even European colonies remain disturbed for up to a week after collection (see Figure 7.5) and it is said by Mitev (1971) that colonies from which venom has been collected every three days produce 14% less honey. Morse and Benton (1964b) found no such evidence for reduced productivity, however. Galuszka (1972) found that when using electro-shock treatment, the most efficient collection cycle was three 15-minute stimulations at intervals of three days, repeated after 2 - 3 weeks. An Argentinean beekeeper found that by modifying the electric stimulus, his collection efficiency greatly increased and the bees remained disturbed for less time.

The various trap designs stimulate bees by applying a mild electric shock through wires above the collecting tray. The most widely-used designs are modifications of the one first presented by Benton et al., (1963). A review by Mraz (1983) discusses further developments. The trays are placed either between the bottom board and brood chamber at the hive entrance (see Figure 7.3) or in a special box between supers and the hive cover, (Palmer, 1961, USA Patent 3,163,871, 1965, as cited by Crane, (1990).

a)
Figure a) Mr. Mraz with an electro-shock venom collector in his beeyard.

b)
Figure b) Placing the  collector in front of the hive entrance. (Courtesy of B. Weeks)

Figure 7.3 a) Mr. Mraz with an electro-shock venom collector in his beeyard.
b) Placing the  collector in front of the hive entrance. (Courtesy of B. Weeks)

When shocked, bees sting the surface on which they are walking. In some traps, this may be a glass plate or a thin (0.13 mm thick) plastic membrane, nylon taffeta or silicon rubber under which a collecting plate (preferably made of glass) or absorbent tissue receives the venom (see Figure 7.4). Venom dries rapidly on glass plates and can be scraped off with a razor blade or knife. Absorbent tissue is washed in distilled water to extract the venom, which then should be freeze-dried. Collection on glass is generally easier and produces a product which is easier to store, ship and process. During handling of dry bee venom, protective gloves, glasses and dust masks should be worn to avoid any contact with, or inhalation of the highly concentrated venom.

It is unlikely that a bee will eject all the contents of its venom sac, even after repeated stinging. Therefore, typically, only 0.5 to 1.0 jil (0.2 júl - Crane, 1990) of venom can be collected per bee, with an average of ten stings per bee (Mu~ller, 1939 and O'Connor et al., 1967). This results in less than 0.1 ijg (0.11 jig - Crane, 1990) of dry venom per bee. Consequently, at least 1 million stings are required to make one gram of dry bee venom. Dotimas and Hider (1987) report that 1 g of venom can be collected from twenty hives over a two hour period.

Close-up of collecting device with stings.

Figure 7.4 : Close-up of collecting device with stings. The steel wires are approximately 6 mm apart and suspended 1 to 3 mm above the thin silicon rubber film or directly above the glass plate in other models. The wires are alternately grounded and charged to a maximum of 33 volts. A lower voltage is effective, too. Preferably a collecting surface should be used which does not make bees loose their sting. (Courtesy of B. Weeks)

Instead of collecting bee venom, adult bees may be used to sting the patient directly. This is the way to apply the venom in its freshest, most complete and cheapest form. To collect the bees, a small hole is made in the brood chamber, super or inner cover. To avoid colony disturbance, the hole is opened and a collecting jar placed over it until a sufficient number of bees have come out. Small groups (10-100) of workers can be maintained at home for up to 2 weeks. They should be kept in the dark, in a small box (with one side made of fly-screen) and with access to sugar syrup. Care needs to be taken to keep ants away. Alternatively, bees can be collected from frames or the hive entrance by a suction device similar to the one described in Figure 6.6. However, a screen should be placed over the tube leading to the mouthpiece to prevent any bees from reaching the mouth.

7.7 Venom products

Bee venom may be sold as whole bee extract, pure lidquid venom or an injectable solution, but in either form the market is extremely limited. Most venom is sold in a dry crystalline form.

Honeybees outside of the hive shortly after electro-shock treatments.

Figure 7.5: Honeybees outside of the hive shortly after electro-shock treatments. The venom extraction board is still leaning over the hive entrance. (Courtesy of B. Weeks)

Since venom does not need to be processed, it can be prepared wherever bee venom therapy finds sufficient support. Production in small quantities is easy, as long as stringent sanitary controls and aseptic working conditions can be provided. The beekeeper has to work under extremely clean conditions, since most of the venom preparations will later be used for injections into humans or animals.

For injections, the venom can be mixed at the time of injection with injectable fluids, such as distilled (sterile) water, saline solutions and certain oils, or it may be taken from prepared ampoules. Ampoules with set doses of ready-to-inject venom should only be prepared by certified pharmaceutical laboratories, because of the need to maintain stringent aseptic conditions and to measure the dosages very precisely.

There are creams available which include bee venom (e.g. Forapin and Apicosan in Germany, Apivene in France and Immenin in Austria) which are used for external application on arthritic joints (BeeWell, 1993 and Sharma and Singh 1983) but neither the ingredients nor their proportions are known to the author. A general recipe for bee venom ointments is given in section 7.13.

Tablets can be impregnated with quantities of bee venom, but Sharma and Singh (1983) recommended the removal of toxic proteins, such as Melittin and the use of colours to indicate different dosages. The tablets should be placed under the tongue, but no indication is given to the effect or usefulness of such a preparation.

Some specialized laboratories may be able to separate and purify different venom compounds and sell them to scientific and pharmaceutical laboratories. Phospholipase A2 and highly active peptides are among some of the proteins purified from bee venom for scientific suppliers or laboratories (Schmidt and Buchmann, 1992). Entry to this limited market requires a highly sophisticated laboratory and very well-trained technicians and chemists.

No further use or inclusion of venom in other products is presently known to the author.

Though not directly related, bee sting emergency kits can be sold in some countries, particularly to people who are allergic. They also should be at hand for any beekeeper working with Africanized honeybees and at training centres, police and fire departments, in areas with Africanized honeybees. In the USA, they are now available only against a prescription. Such a kit (e.g. Anakit by Hollister Stier, USA) should contain at least:

1) One syringe with a premeasured content of epinephrine (adrenaline) or atropine, for immediate intramuscular injection - usually 0. 3m1 of a diluted solution of epinephrine (1:1000) in saline solution. There are special, easy-to-use, syringes available from bee supply houses or through pharmacies, which can even be used through clothing (Epipen by Centre Laboratories, USA).

2. anti-histamine tablets.

3. tourniquet.

4. instructions about when, where and how to use the syringe and anti-histamine tablets; when not to use epinephrine, and where to seek medical help.

Epinephrine injections should be given only in extreme emergencies when no other medical help is available. The sting emergency kit has a limited shelf-life and should be kept refrigerated when not in use.

7.8 Buying

The best way to buy bee venom is in the crystallised form, since it is more stable, impurities are easier to detect and adulteration is less likely. The colour of both crystals and powder should be a very light yellow.

Liquid venom as mentioned in section 7.2 should be clear and colourless. Darker venom is slightly oxidized and may have lost some of its effectiveness.

As with all other products where immediate testing is not possible or is very expensive, the producer should be one who is well-known and who can be trusted to produce a high quality product. The producer as well as the buyer should have adequate storage facilities.

7.9 Storage

Even dried bee venom should be stored refrigerated or preferably frozen and it should always be kept in dark bottles in the dark. All producers and buyers should closely observe these conditions. Dried bee venom can be kept frozen for several months, but should not be kept refrigerated for more than a few weeks. Liquid venom and diluted venom can be stored for similar periods if maintained in well sealed, dark glass containers.

7.10 Quality control

There are no official quality standards, since bee venom is not recognized as an official drug or as a food. Purity analysis may be carried out by quantitative analyses of some of its more stable or more easily measured components such as melittin, dopamine, histamine, noradrenaline or those for which contamination is suspected.

A nematode, Panagrellus redivivus was reported to react selectively and specifically to bee venom and a quantitative analysis of the venom in pharmaceutical preparations was developed by Tumanov and Osipova (1966) using this organism.

Pence (1981) describes a method for testing the biological activity of bee venom by measuring electric pulses from muscles of excised honeybee abdomens in response to the volatile materials from bee venom.

Guralnick et al., (1986) described standardization and quality control methods for purity and effectiveness of Hymenoptera venom, including honeybee venom.

7.11 Caution

Collecting bee venom requires careful work with the highest degree of cleanliness, since the venom will be injected directly without further processing or sterilization. Protection of the collector against the disturbed bees and highly irritative dry venom is very important, too. Since people up to several hundred meters away might get stung by the highly irritated bees, further precautions at the time of collection in the apiary must be considered.

When handling dry venom, laboratory gowns, gloves and face masks should be worn to avoid getting venom dust into the eyes and lungs. All equipment should be carefully washed afterwards. Contact between other people and contaminated material should be avoided People who do not regularly handle bees, who only get stung occasionally or are exposed occasionally to venom dust, run the risk of developing allergies.

Using bee stings for self-treatment of various diseases can be risky, because allcrgies to bee venom can be developed quickly even after long periods of use. An emergency kit (see section 7.7) or quick access to an emergency service should always be available. No other side-effects have been reported, but regular supervision, check-ups and controls should be continued with competent doctors trained in apitherapy.

Since severe allergic reactions are possible, bee venom should not be casually included in any health or medicinal products. Products containing bee venom need labels stating the contents and warning people of possible allergic reactions.

7.12 Market outlook

Bee venom is a highly specialized product with only very few buyers. The market volume is relatively small too, although there are no comprehensive surveys. The main venom producer in the USA has produced only about 3 kg of dry venom during the last 30 years (Mraz, 1982) but there is a large producer in Brazil and more or less significant amounts are produced in many other countries.

Prices in 1990 varied greatly between US$100 and US$200 per gram of dry venom (Schmidt and Buchmann, 1992). Prepared for injections or sold in smaller quantities, prices can be much higher. However, the beekeeper often does not get this price. The prevailing prices in European and Asian markets are generally slightly lower.

Local manufacturing of the pure venom however, is relatively easy and within the means of many beekeepers; no expensive or high technology processing is required except refrigeration, but its economic  feasibility depends on access to the few specialized buyers. In contrast, the venom in less  controlled dosages is available almost everywhere, from a beekeeper or one's own hives, free or at very low cost. Often, the only price is the life of the bee.

Though the fractionation of venom goes beyond the means of a small local enterprise, several people working in the field feel that, with further research, there will be a small market niche for specialized laboratories. Since there are several pharmacologically interesting substances in bee venom and since apitherapy may become officially accepted in the future, a better market for the whole product or for special fractions might develop. However, much depends on the official acceptance of bee venom therapy.

7.13 Recipes

Ointments can be prepared by thoroughly homogenizing bee venom with white Vaseline, petrolatum or melted animal fat, and salicylic acid, in the ratio of 1:10:1. The salicylic acid softens the skin, increases its permeability and is a treatment for rheumatism even on its own. The ointment may contain a small amount of silicate crystals to act as an abrasive (Sharma and Singh, 1983).

Other preparations consist of mixing bee venom with sterile, injectable fluids and packaging them in single dosages in glass vials or syringes. In some packages the dry venom is kept separate from the fluid and the two are mixed when the vial is broken.

Techniques for separating the different compounds in venom are far beyond the scope of this book. Such information can be obtained from properly trained chemists.

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