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The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
1
The Bee Venom Book
Eros, stung by a bee,
Ran away and cried for plea:
Venus, mother, I cry ,
Please help me or I‘ll die
What a terrible disgrace –
A dragon bit me on my face
Venus comforting her son
Speaking with a mocking fun -
The little bee's tiny sting
Is for you an earnest thing
But more painful and real hard
Are your stings in human’s heart
Anacreontean songs, 6 BC
Venus, Eros and the bees
By A. Dürer, 1514
Be aware that this online book is only for private use and should not be copied and reprinted as some of the images
are not copyrighted.
I would appreciate your feedback at info@bee-hexagon.net
Stefan Bogdanov, Muehlethurnen, Switzerland
The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
2
Bee Venom:
Production
Composition
Quality
Stefan Bogdanov
BEE VENOM PRODUCTION
Bees produce their venom in their venom glands,
schematically described in the figure left. The BV is
secreted in a branched acid gland (above) and in the
alkaline Dufour’s gland (below), in the whole BV both
secretions are mixed.
New born bees do not sting. Venom synthesis begins
after two or three days, while the maximal production
rate is reached when bees are two to three weeks old.
Older worker bees produce less venom. One sting
contains about 100 µg of dry BV.
Drones do not have stings, while bee queens have BV,
its maximum quantity being that of newly emerged
queens, in order to facilitate their fight for survival
against competing queens.
The sting consists of three parts: a stylus and two
barbed slides (or lancets), one on either side of the
stylus. The bee does not push the sting in but it is
drawn in by the barbed slides. The slides move
alternately up and down the stylus so when the barb of
one slide has caught and retracts it pulls the stylus and
the other barbed slide into the wound. When the other
barb has caught it also retracts up the stylus pulling the
sting further in. This process is repeated until the sting
is fully in and even continues after the sting and its
mechanism is detached from the bee's abdomen.
The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
3
When stinging a mammal the sting is barbed so that it
lodges in the victim's skin, tearing loose from the bee's
abdomen and leading to its death in minutes. However,
this only happens if the victim is a mammal or bird.
It has been speculated why bees lose their stings. The
first theory says that the sting evolved early in
evolution, before the appearance of mammals.
Another theory says that both bees and mammals
evolved at the same time. Losing the sting and dying
during the fight with mammals is hypothesised to be
more advantageous than the eventual loss of several
worker bees because the bee can deliver a greater
amount of BV into the victim. The alternative is the life
loss of several worker bees
14
All images were freely available in Internet, the sources could not identified
HARVEST OF BEE VENOM
Today BV is collected for commercial purposes by special collectors.
Collection of bee venom
Scheme of a BV collector, (after U. Mueller)
Most commercial venom collectors are composed of four
parts:
- Battery or accumulator (24 to 30 V)
- Transformer from constant to alternating current, with
impulse frequency of 50 to 1000 Hz and an impulse
duration of 3 to 6 seconds.
- Collector frame consisting of an electric wire net and a
glass plate, covered by a thin polyethylene membrane.
The collector can be mounted in or out of the hive. The
bees get in contact with the charged wire net and are
stimulated to sting through the membrane and spray their
venom on the glass plate. The glass plates are dried in a
dark, well ventilated room.
Commercial BV collector
Beekeeper with a BV collector
The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
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An electric BV collecting technique was described for the first time in 1954 by Markovic und Mollnar
15
. Many
different models have been developed
4, 6, 13, 17, 21
. The collectors have been used under different conditions:
• Voltage: from 24 to 30 V,
• Impulse duration: 2-3 seconds
• Pauses: 3 to 6 seconds; impulse frequency from 50 to 1000 Hz.
Bees are not harmed during the BV collection. Repeated 3 hours collection periods, carried out 3 to 4 times per
month do not harm bees, resulting in a total harvest of 4 g dry BV. This collection resulted in a decrease of brood
production and honey yield of about 10-15 %. If the collection was less frequent, e.g. 3-4 times per season, the bee
performance was not influenced
13
. 10’000 bees are needed for the collection of 1 g dry BV
23
.
BV is gathered commercially in Eastern Europe, Far East and North and South America.
BV from different honeybee species are slightly different but its overall activity is similar
12
In warm and humid zones the BV can be more toxic than in cold temperate zones. Regarding the BV in different
Apis types, it was found out that A. melifera and A.dorsata venom had similar toxicity, while A. dorsata venom
contains much more alarm pheromones
23
.
Different BV components can be isolated for special uses in medicine and biology.
PROPERTIES AND COMPOSITION
Properties of fresh bee venom, after
3, 5, 20
:
A bee venom drop
• The water content varies between 55 and 70 %.
• Yellowish opalescent liquid sometimes almost
colourless,
odour: honey-like; taste: aromatic, bitter, acidic and
hot
• Soluble in water and diluted acids, insoluble in
alcohol
• pH: 4.5-5.5
• Specific gravity: about 1.13
• Soluble in water, about 10 % are insoluble, water
solutions are unstable, insoluble in ethanol,
dried BV: a gum like powder
The collected venom dries quickly in ordinary room
temperature, turning into a yellow-brownish powder
crystalline mass.
BV, relatively stable, destroyed by sun light and higher
temperatures, stable at low temperature. easily destroyed by
oxidizing substances: potassium permanganate, potassium
sulphate; halogen elements-chlorine and bromine-destroy it
very quickly; the effect of iodine is much slower. Alcohol
possesses a strong and quick destructive effect on the
venom. In contact with tincture of iodine, the alcohol is
more destructive than the dissolved iodine,
after
3, 19
The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
5
Composition
Bee venom is a complex mixture of proteins, peptides and low molecular components. Nowadays its components
have been characterised. The main components are proteins and peptides. The different components are reviewed
recently
1
. Here the most relevant components are given.
The composition of dry BV is given in the table below. The composition of fresh and dried BV differs mainly in
regards to the volatile components; the overall biological activity is similar.
Proteins (Enzymes)
The enzymes are proteins catalyzing specific reactions. There are 5 enzymes in BV.
Polypeptides
Polypeptides are smaller in molecular weight than enzymes, made of 2 or
more amino acids. BV has numerous polypeptides (see table 1), the main
one being melittin, which is also the main component of BV. Melittin has a
MW of 2840 daltons but it can reach 12 500 daltons because it can be also in
a tetrameric form
7-9
The protein and the melittin electrophoretic patterns are typical of the
honeybee species
12
.
To the left: the structure of melittin (source: Wikipedia)
Low molecular compounds
BV contains smaller quantities of low molecular compounds are different in nature: amino acids,
catecholamines, sugars and minerals. Sugars have been identified in some BV preparations, but if BV is
collected with a collector preventing the contamination by pollen and nectar, it does not contain
carbohydrates
20
.
Table 1: Composition of bee venom dry matter,
after
1, 2, 5, 20, 23
Substance Group Component % of dry weight
Proteins (Enzymes) Phospholipase A2
Phospholipase B
Hyaluronidase
Phosphatase
α - Glucosidase
10-12
1
1-2
1
0.6
Peptides Melittin
Apamine
MCD peptide
Secapine
Pamine
Minimine
Adolapine
Procamine A, B
Protease inhibitor
Tertiapine, cardiopep, melittin F
40-50
2-3
2-3
0.5-2
1-3
2
0.5-1
1-2
0.1-0.8
1-2
Phospholipids 1-3
Biogenic amines Histamine
Dopamine
Noradrenalin
0.5-2
0.2-1
0.1-0.5
Amino acids Aminobutyric acid, α-amino acids 1
Sugars Glucose, fructose 2-4
Volatiles (pheromones) Complex ethers 4-8
Minerals P, Ca, Mg 3-4
The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
6
QUALITY OF BEE VENOM
If it is not protected, oxidation will change the color from white to brownish-yellow. Changes caused by oxidation
of certain components of the venom may decrease its healing effect. There are different kinds of venom such as:
pure whole dried, whole dried and freeze-dried (lyophilized) BV. Pure whole dried BV is the purest venom. It is
white in colour (often it is snow white), not contaminated with foreign materials and colourless when it is used in a
solution. Optimal quality of BV can be achieved when it is harvested correctly. Contamination with bee faeces,
dust, pollen, honey and other bee hive components should be avoided.
Freeze-dried BV is highly processed and purified venom. During the preparation its moisture content and any other
contaminants are removed in order to purify and preserve it. Some of the active components may be removed also
if an uncontrolled purification method is used. It is widely used in creams, liniments and ointments. In a tablet
form, it can be used to prepare venom solution for electrophoresis or phonophoresis (ultrasound) applications. It is
easy to sterilize by syringe filtration
22
.
If BV is protected from moisture and light it can be stored for five years or more. It will not lose its toxicity,
however its healing effects are reduced by storage. Freeze-drying is the most effective method of preserving BV.
Optimal quality of BV can be achieved when it is harvested correctly. Contamination with pollen, honey
and other bee hive components should be avoided.
There is no international standard for BV quality. However, following quality criteria are followed by
pharmaceutical firms:
Table 2: Suggestion for a quality standard for dry bee venom, according to the literature and
after
16
,
20
Quality criteria Requirement
Organoleptic properties: typical
2 % BV solution: Extinction at 420 nm smaller than 0.55
Water content: less than 2 %
Water-insoluble substances less than 0.8 %
Sugars less than 6.5 %
Mellitin by HPLC
Biological activity of hyaluronidase,
phospholipase, melittin, protease-inhibitor satisfactory
Radio-immuno tests satisfactory
Toxicity LD
50
3.7 ± 0.6 mg/kg*
* - LD
50
– intravenous injection of a dose producing 50 % of mice survival
Russian bee venom standard
ФС 42-2683-89
Quality criteria Requirement
Organoleptic properties: typical
Water content: less than 12 %
Water-insoluble substances less than 10 %
% solids in the sol less than 2
Hemolysis time less than 480 seconds
Phospholipase activity units less than 100
Hyaluronidase activity units more than 70
The Bee Venom Book, Chapter 1
Bee Product Science, www.bee-hexagon.net, April 2016
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Melittin and Apamin should also be determined in a future standard as they are the main biologically
relevant components. Determination of melittin by HPLC in dry commercial BV in Iran showed variation
between 8 and 51 %
11
or in a later study by the same authors it was measured in bee-hive collected BV in
Iran, between 21.9 and 66.4
10
In a Chinese study with HPLC-DAD-MS/MS do determine melittin and apamin values of 46- 53 % and
2.2-3.7 % were found for both components (% of total dry weight)
24
In Poland melittin and apiamin were determined by HPLC and following values were found in % of total
dry weight: melittin: 61-70 %, apamin: 2.1 – 4.2
18
PRODUCTION AND TRADE
BV is produced in many countries, mainly in Eastern Europe, South East Asia and the Americas. There are no
official figures on the quantity of BV traded. Most of the BV is used for apitherapy and for desensitation in
hospitals, but lately there is demand for the cosmetic industry.
Good commercial source of whole BV are www.beevenom.com and
www.beevenomlab.com
Some companies produce and offer bee venom components such as melittin and apamin.
References
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2. BANKS, B E C; SHIPOLINI, R A (1986) Chemistry and pharmacology of honey-bee venom., In Piek, T (ed.)
Venoms of the Hymenoptera, Academic Press; London; pp 330-416.
3. BECK, B F (1935) Bee venom therapy. D. Appleton-Century Company New York and London
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