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Propolis

Propolis

Propolis is a mixture of various amounts of beeswax and resins collected by the honeybee from plants, particularly from flowers and leaf buds. Since it is difficult to observe bees on their foraging trips the exact sources of the resins are usually not known. Bees have been observed scraping the protective resins of flower and leaf buds with their mandibles and then carrying them to the hive like pollen pellets on their hind legs. It can be assumed that in the process of collecting and modelling the resins, they are mixed with some saliva and other secretions of the bees as well as with wax.


Figure 5.1 : Honeybees frequently use propolis to reduce the size of the entrance for better defence.

 
These resins are used by worker bees to line the inside of nest cavities and all brood combs, repair combs, seal small cracks in the hive, reduce the size of hive entrances (see Fig. 5.1) seal off inside the hive any dead animals or insects which are too large to be carried out and perhaps most important of all, to mix small quantities of propolis with wax to seal brood cells. These uses are significant because they take advantage of the antibacterial and antifungal effects of propolis in protecting the colony against diseases. Propolis has been shown to kill the bee's most ardent bacterial foe, Bacillus larvae - the cause of American Foul Brood (Mlagan and Sulimanovic, 1982; Meresta and Meresta, 1988). The use of propolis thus reduces the chance of infection in the developing brood and the growth of decomposing bacteria in dead animal tissue.

The composition of propolis depends on the type of plants accessible to the bees. Propolis changes in colour, odour and probably medicinal characteristics, according to source and the season of the year. Moreover, some bees and some colonies are more avid collectors-generally to the dismay of the beekeeper, since propolis is a very sticky substance which, in abundance, can make it difficult to remove frames from the boxes.
Foraging for propolis is only known with the Western honeybee Apis mellifera. The Asian species of Apis do not collect propolis. Only Meliponine or stingless bees are known to collect similarly sticky resinous substances, for sealing hives and constructing honey and pollen pots for storage. In this bulletin, however, propoli shall refer only to resins collected by honeybees, since almost all of the research has been done on it. There may well be similar traditional uses for resins collected by Meliponids.

In the natural distribution ranges of Apis mellifera, a multitude of traditional uses are known for this versatile substance. The Greeks and Romans already knew that propolis would heal skin abscesses and through the centuries its use in medicine has received varying attention. The ancient Egyptians knew about the benefits of propolis and in Africa it is still used today, as a medicine, an adhesive for tuning drums, sealing cracked water containers or canoes and dozens of other uses. It has been incorporated in special varnishes such as those used by Stradivarius for his violins (Jolly, 1978).
An excellent review in Spanish on the production, characteristics and uses of propolis was published by Asis (1979 and 1989) another good overview (in English) was APIMONDIA (1978). A brief, more recent review in English is presented by Schmidt and Buchmann (1992).
Physical characteristics of propolis

The colour of propolis ranges from yellow to dark brown depending on the origin of the resins. But, even transparent propolis has been reported by Coggshall and Morse (1984).

At temperatures of 250 to 45 0C propolis is a soft, pliable and very sticky substance. At less than 150 C, and particularly when frozen or at near freezing, it becomes hard and brittle. It will remain brittle after such treatment even at higher temperatures. Above 45 0C it will become increasingly sticky and gummy. Typically propolis will become liquid at 60 to 700C, but for some samples the melting point may be as high as 1000C.
The most common solvents used for commercial extraction are ethanol (ethyl alcohol) ether, glycol and water. For chemical analysis a large variety of solvents may be used in order to extract the various fractions. Many of the bactericidal components are soluble in water or alcohol.
The composition of propolis

In one recent analysis of propolis from England, 150 compounds were identified in only one sample (Greenaway, et al., 1990), but in total more than 180 have been isolated so far. It appears that with every new analysis, new compounds are found.

Propolis resins are collected from a large variety of trees and shrubs. Each region and colony seems to have its own preferred resin sources, which results in the large variation of colour, odour and composition. Comparisons with tree resins in Europe suggest that, wherever Populus species are present, honeybees preferably collect the resins from leaf buds of these trees.

A Cuban study suggests that the plant resins collected are at least partially metabolized by bees (Cuellar et al., 1990). The presence of sugars (Greenaway et al., 1987) also suggests some metabolization by bees, i.e. as a result of adding saliva during both scraping and chewing.
A list of the major classes of chemicals occurring in propolis is given below with references to some recent reviews and analyses from different countries (Table 5.1). The major compounds are resins composed of flavonoids and phenolic acids or their esters, which often form up to 50% of all ingredients. The variation in beeswax content also influences the chemical analysis. In addition it must be said that most studies do not attempt to determine all components, but limit themselves to a class of chemicals or a method of extraction. The selection of the studies presented here is based on the most recent publications with preference given to the most complete studies or to studies from countries where these are the only references.

The physiological effects of propolis
Unconfirmed circumstantial evidence
The following uses of propolis or its extracts have been found in literature, but without substantiating evidence or reference to scientific studies:

anti-asthmatic treatment in mouth sprays,

support of pulmonary system,

anti-rheumatic (Donadieu, 1979),

inhibition of melanoma and carcinoma tumour cells,

tissue regeneration,

strengthening of capillaries,

anti-diabetic activity,

phytoinhibitor,

inhibiting plant and seed germination (Donadieu, 1979) in general and potato and leaf salad seed germination (Bianchi, 1991) in particular.

Table 5.1:
The major compounds of propolis as analyzed in recent publications.

Scientific evidence
One of the most widely known and extensively tested properties of propolis is its antibacterial activity. Many scientific tests have been conducted with a variety of bacteria, fungi, viruses and other microorganisms. Many of the tests have shown positive control of the organisms by various extracts and concentrations of propolis. A synergistic effect has been reported for propolis extract used together with antibiotics (Chernyak, 1971). Whether propolis exhibits bactericidal or bacteriostatic characteristics often depends on its concentration in the applied extract. Sometimes, propolis extracts are more effective than commercially available drugs (Millet-Clerc, et al., 1987). In all cases, the specific conditions and extracts have to be closely considered. Proven effects of propolis on microorganisms are listed in Table 5.2.

Though there is a large variety of effects attributed to propolis, many of the reports are based on preliminary studies. If clinical trials were conducted, they were rarely based on large numbers of patients or rigorous test designs such as the double-blind placebo test (Table 5.3). The majority of the studies were conducted in East European countries. Much practical work and research is also being done in China, but information is difficult to obtain, not least because of the language barrier. Western European and North American medical research has largely ignored this source of milder and widely beneficial material. More detailed studies are warranted to determine the potential benefits from the medicinal use of propolis, particularly for intestinal, dermatological and dental applications.

In addition to the selected studies cited here, there have been over 500 publications in the last 18 years alone. Most were in vitro studies, but clinical trials were also conducted. These can be researched by those further interested in the uses of propolis in the collection of abstracts prepared by IBPA which is available from them.

The uses of propolis today
In cosmetics
Dermatological and cosmetic applications are at this time probably the most common uses for propolis and its extracts (Lejeune, et al., 1988). Its effects on tissue regeneration and renovation have been well studied. Together with its bactericidal and fungicidal characteristics it provides many benefits in various applications in cosmetics. For some recent specific references on scientific studies, the reader should refer to the section on the effects of propolis (5.4.2). More detailed information on practical application of propolis in cosmetics can be found in Chapter 9.

In medicine
General medicinal uses of propolis include treatment of the cardiovascular and blood systems (anaemia), respiratory apparatus (for various infections), dental care, dermatology (tissue regeneration, ulcers, excema, wound healing - particularly burn wounds, mycosis, mucous membrane infections and lesions), cancer treatment, immune system support and improvement, digestive tracts (ulcers and infections), liver protection and support and many others. Some references to these applications can be found in the list of scientifically proven effects of propolis otherwise one might refer again to IBRA's collection of abstracts, Apimondia and the American Apitherapy Society.

Direct external application of ethanol extracts or concentrated ointments (with up to 33% propolis) have given good results in veterinary use for wound healing and sores. Plastic surgery too, is using propolis extracts for improved wound healing and reduced scar tissue development.

Traditional use
In Europe and North Africa, the special wound healing properties of propolis were already known to the Egyptians, Greeks and Romans and in ancient times. In records of the 12th century, medicinal preparations with propolis are described for treating mouth and throat infections, as well as caries. Propolis probably has been more commonly used in wood preservatives or varnishes than may be suggested by the single, frequently cited reference to Stradivarius (Jolly, 1978).
In sub-Saharan Africa, propolis is still used today in herbal medicines and the more mundane applications mentioned earlier such as waterproofing containers and wood, adhesive, bow string preparation and for tuning drums.

Food technology
The antioxidant, antimicrobial and antifungal activities of propolis offer scope for applications in food technology. One special advantage is that, unlike some conventional preservatives, the residues of propolis seem to have a generally beneficial effect on human health. However, only very few studies have been done on the possible side-effects of increased consumption of propolis. Individually, some of the components identified in propolis can be very damaging to human health.
Mizuno (1989), registered a patent which includes propolis as a preservative in food packing material.
Extension of frozen storage life of fish by 2-3 times is cited including Donadieu (1979), but without reference to original studies. propolis is permitted as a preservative for frozen fish. by various authors, In Japan, the use of Addition of only 30 ppm (parts per million) of propolis to the rations of laying hens increased egg production, food conversion and hen weight by S to 6% (Bonomi, et al., 1976). Ghisalberti (1979) reports additional weight gains for broiler chicken of up to 20% when 500 ppm of propolis was added to their diets.

Others
The search for new uses of propolis continues. Sangalli (1990) mentioned use of propolis for post-harvest treatment and conservation of fruits. Applications in pesticides and fungicides are still in the testing phase. However, for many of its traditional uses propolis is being replaced by more readily available, sometimes more effective but often also more toxic alternatives.
Beekeepers use propolis, melted together with wax or in an ammonia solution (Anon, 1982) to apply to the inside of hives or swarm traps to attract swarms. Adequate ventilation and aeration after painting with the ammonia solution are both necessary. Rubbing propolis or painting it (after melting with wax from old combs) works as well or better and avoids the use of noxious and toxic ammonia.
The current trend to return to environmentally safer and less energy intensive production methods in many developed countries, the increased buying power of consumers and growing markets for more expensive products may lead to considerable growth in the use and new applications of propolis, particularly in cosmetics and food technology.

Caution
Hausen et al., (1987) cited almost 200 cases in which people have shown allergic reactions to propolis. In some cases of direct contact with propolis, this may have also been a result of contamination by other bee products such as pollen or bee hairs. However, extracts and products containing propolis extracts have been shown to cause allergic reactions as well (Hausen, et al., 1987, Hausen and Wollenweber, 1987 and Ko~nlg, 1988) mostly in the form of contact dermatitis. Hashimoto et al., (1988) identified caffeic acid and its derivatives as the major allergenic agents.
Therefore, with all preparations intended for human or animal luse, small quantities should be tried during the first days, slowly increasing to the full dosage (half for children) in order to test for the compatibility of the preparatino or allergic reactions. Equally, termination of medical treatments prescribed by a physician should be gradual, slowly reducing the daily dosage.

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