Bee Venom, Propolis, Pollen and Royal Jelly Processing Part TWO

Continued from Part ONE

c) Royal Jelly Collection

Royal jelly is produced by stimulating colonies to produce queen bees outside the conditions in which they would naturally do so (swarming and queen replacement). It requires very little investment but is only possible with movable comb hives. Expert personnel are required, who are able to devote considerably more time than is commonly required for the production other bee products. Without this prerequisite it is possible to only occasionally collect the contents from cells of natural swarms-and this amounts to no more than a gram or two per hive.

The basic requirements are movable comb hives, preferably some queen excluders, queen cups(made from wax or plastic), a transfer needle, a spoon or suction device to remove royal jelly, dark glass vials and a refrigerator. Special hive modifications may be used for large scale production. Feeding with sugar syrup (1:1 in sugar/water) increases cell acceptance, even when flowers are available.

Mature queen cells, i.e those with larvae four days old (3 days after grafting), must be brought quickly into the extraction room. The open, narrow part of the cells is cut to facilitate and speed up collection. Then the larvae are removed with a pair of soft forceps, taking care not to harm them and contaminate the jelly. The royal jelly is extracted by emptying each cell with a small spatual, by sucking it up with a special moth operated device, with a pump operated device or by centrifugal extraction. Following extraction the cells are immediately ready for another rearing cycle.

The royal jelly must be filtered using a fine nylon net (nylon stockings are excellent) to eliminate fragments of wax and larvae. Metal filters should not be used. The jelly should be placed into dark glass vials or food-grade plastic containers, avoiding any excessive exposure to air. It should be refrigerated immediately. Any material or equipment contacting royal jelly-including hands-must be clean and disinfected using heat or pure alcohol. The laboratory must be kept impeccably clean and extraction should never be done outside or in sunlight.

The commercial production of royal jelly requires a methodical approach, good organization and precise timing. Constant attendance is essential as one day off can eliminate two days of production. In order to have a weekly day of rest (e.g Sunday) no queen cells would be introduced on Thursday, which means that there will also be no collection on the following Wednesday.

These techniques are suitable for both small and quite large enterprises. Depending on the intended market, the approach can be either one of low cost or one in which all collecting, Processing and distribution takes place in highly controlled environments. The latter will result in a product which is better suited for industrial use.

Royal jelly has a limited shelf-life. Early beliefs in the extreme instability of royal jelly activity, based on the alleged rapid loss of the “queen determination” factor have not been confirmed. Since neither the mode of activity nor the actual effects of royal jelly are known, there are no data available on changes in its biological effectiveness on human after long term storage.

Since royal jelly is an emulsified product and not cellular tissue, freezing presents no particular problem and common household freezers can be used.

Like all other bee products, royal jelly has its own microbiological protection and presents few microbiological storage problems when it is in its natural state. This protection however is not absolute and certain hygiene precautions must be observed during production and storage. Hygienic working conditions and clean containers are a minimum requirement, and airtight containers should be used to provide additional protection not only against contamination but also against oxidation.

d) 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. Venom collected from surgically removed sacs showed different protein contents from the collected with the electroshock method.

Venom collection apparatus for placement at different levels within a segmented bee colony structure, said apparatus comprising:

a) A housing member having a front wall, opposing, sidewalls and a back wall and being dimensioned to fit within the segmented colony structure:

b) A frame member removable mounted in said housing and having a central opening therein to permit the movement of bees there through.

c) An array of spaced electrical conductors extending across the opentining in said frame member, said conductors being spaced to permit the passage of bees there through:

d) First access means opening adjacent to at least one wall of said housing member and positioned beneath said frame member to provide an external access path for bees in the colony:

e) Means for establishing a voltage difference between adjacent conductors in said array whereby bees passing through said array from said first access means encounter adjacent conductors and are eclectically stimulated and

f) Collection means removably mounted in said housing member for withdrawal there from and located beneath said first access means and external access path, said collection means receiving venom from bees excited by contact with adjacent conductors in the spaced overlying from member.

The various trap designs stimulate bees by applying a mild electric shock through wires above the collecting tray.

When shocked, bees sting the surface on which they are walking. In some traps, this may be a glass plate or a thin (0.13mm thick) plastic membrane, nylon taffeta or silicon or silicon rubber under which a collecting plate (preferably made of glass) or absorbent tissue receives the venom. 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 of venom can be collected per be, with an average of ten stings per bee. This results in less than 0.1 ijg (0.11 jig-Cranes, 1990) of dry venom per bee. Consequently, at least 1 million stings are required to make one gram of dry bee venom.

Collecting be venom requires careful work with the highest degree of cleanliness, since the venom will be injected directly without further processing of sterilization. Protection of the collector against the disturbed bees and highly irrigative 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.

Bee Venom, Propolis, Pollen and Royal Jelly Processing Part ONE

Honey bees are the sole sources of honey and bees wax with usual qualities. Honey bees also produce propolis, a gummy substance made from tree sap that has anti-bacterial properties, and royal jelly and pollen for human consumption. Honey bee venom is extracted for the production of anti-venom therapy and is being investigated as a treatment for several serious diseases of the muscles, connective tissue, and immune system, including multiple sclerosis and arthritis

The production process consists of the following operations:

a) Pollen collection

b) Propolis collection

c) Royal jelly collection

d) Venom collection

a) Pollen Collection

Extreme care should be taken that pollen is not contaminated by bees collecting from flowers treated with pesticides. During, and for several days or weeks after treatment of fields or forests in an area of several square kilometers (in a circle of at least 3-42 km diameter) around the apiary, no pollens should be collected. This is independent of the method of pesticide application. Even systemic pesticides have been shown to concentrate in pollen of, for example coconut. Since a pollen pellet is collected from many flowers, even small quantities of pesticides per flower can be accumulated rapidly to reach significant concentrations.

Though pollen pellets are collected before they enter the hive, treatment of colonies for bee diseases, can contaminate the pollen pellets. Though, for example, cleaning of debris from the hive and bees regurgitating syrup, nectar or honey during collection of the pellets.

Pollen pellets are removed from the bees before they enter the hive. There are many designs of pollen traps some easier to clean and harvest, others more efficient or easier to install. The efficiency rarely exceeds 50%, i.e. less than 50% of the returning foragers loose their pollen pellets. Bees are ingenious in finding ways to avoid losing their pellets, like small holes or uneven screens and may even rob pollen from the collecting trays, if access is possible. Under some circumstances, pollen collection methods and regimes may interfere with normal colony growth or honey production. Therefore, standard beekeeping manuals should be consulted for the timing of collections.

Pollen should be collected daily in humid climates but less frequently in drier climates. To avoid deterioration of the pollen and growth of bacteria, molds and insect larvae, pollen should be dried quickly. Ants can remove considerable amounts from pollen traps. Studies indicate that losses can be to 30% in temperate climates.

Pollen needs to be dried to less than 10% moisture content (preferably 5% or 8% according to some laws) as soon as possible after harvesting. A simple method uses a regular light bulb so that the pollen does not heat to more than 40 or 45^oc. For solar drying, the pollen suspended high enough above a pollen itself should be covered to avoid direct sunlight and overheating.

After drying, the pollen needs to be cleaned of all foreign matter. A tubular tumbler out of a wire mash with a fan can clean considerable quantities of pollen pellets. Simpler winning methods can be used too.

Since sunlight, i.e. UV radiation, destroys the nutrient value of pollen, other more subtle characteristics probably suffer worse damage. Storage of dry pollen in dark glass containers, or in dark cool places, is therefore a requirement.

Pollen used for cosmetic purposes should have the same, if not a better, quality than destined for consumption as food. The first quality control is assessment of gross contamination with foreign substances, i.e., parts of bee and hive debris. Further controls might include measurement of moisture content and a bacterial count. Determination of various agrochemicals, including drugs used inside bee colonies is possible and may be required in some circumstances. These analyses require sensitive, expensive chromatographic equipment.

b) Propolis collection

The average production of propolis per colony per year has been described as 10 to 300g but the production depends on the bees, the climate, the forest resources and the trapping mechanism. According to personal observations, it may occasionally be considerable higher. If there is any selection by queen breeders and beekeepers, it has been against heavily propolizing bees, since they make work in the apiary more difficult. Bees which produce larger quantities of propolis could be selected if required. 

Contamination of propolis with wax, pieces of wood, paint and other debris should be avoided. The cleanest collection methods employ special traps placed on top of a hive, below the covers or next to lateral walls inside the hives. Thus bees do not mix as much wax with the propolis and no contamination occur during harvesting. Trap harvesting is also faster and may be more productive. 

Traps are basically screens or special plates with small holes which simulate cracks in the hive walls. Bees try to seal the holes and thus fill the trap with propolis. The most economic trap design is an inner cover with a large hole, covered with regular nylon fly screen, secured in place by the points of nails and a perforated frame. The total area exposed by a screen may have to be varied according to the bees and local conditions. Trap harvested propolis usually fetches a better price because of its cleaner and therefore of better quality.

Light, and in particular air circulation are important to stimulate propolis use. Accordingly, traps placed on top of hives should be covered but the hive cover needs to be propped opened slightly to increase air circulation and to allow in some light. In tropical regions it may be necessary to prevent the entry of too much rain. Also, when using a type of bee sensitive to disturbances or likely to abscond, the lid should not be opened too far otherwise bees might escape. Newly established colonies should be given some time to establish themselves before they are used for trapping.

Propolis is removed from traps by cooling the plastic sheets or fly-screens for a few hours in a refrigerator or freezer. Once cooled, the propolis becomes brittle and can be removed from the screens by simply flexing and brushing them, pulling over a table edge or by using a special high pressure air device. The trap is then ready for re-use.

In general, propolis is fairly stable, but proper storage is important. Propolis and its extracts should be stored in airtight containers in the dark, preferably at less than 10^oc-12^oc and away from excessive and direct heat. For similar reasons, very old propolis from the hive should not be mixed with fresher propolis. Over 12 months of proper storage, propolis will lose very little or non of its antibacterial activities. Alcohol extracts may be stored even longer.

c)Royal Jelly Collection 
     See PART TWO

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