6^th Brazilian Encontro and 8^th International Conference on Tropical Bees; Parts I – III

American Bee Journal (May, July, August 2005), Vol. 145

 

By

 

Malcolm T. Sanford

http://apis.shorturl.com

 

In my series on the current status of Brazilian beekeeping I mentioned that the Africanized honey bee’s introduction into that country produced a huge outpouring of scientific study.  The result of this continues to be the legacy of one of the world’s most dynamic faculties involved with tropical bees at the Ribeirão Preto campus of the University of São Paulo.  In 1994, a group of students and faculty at that institution organized the first Encontro (Encounter) to share their tropical bee research results and provide a forum for discussion.  Thus began a series of these meetings taking place every two years. 

According to Dr. Lionel Gonçalves writing in the preface to the sixth Encontro held September 6-10, 2004 in Ribeirão Preto, the faculty never dreamed that the 1994 meeting would grow into one of the most prestigious scientific events in Brazil, and by extension the bee world.  The fourth Encontro held in the year 2000 was especially significant because it honored Brazil’s three bee research giants, Father Jesus Santiago Moure, Dr. Paulo Nogueira Neto and Dr. Warwick Estevan Kerr. 

The recent Encontro commemorated the 40^th anniversary of the Faculty of Philosophy, Science and letters at the University of São Paulo and was combined with the Eighth International Conference on Tropical Bees sponsored by the International Bee Research Association (IBRC).  This coming together of scientists from all around the world resulted in a meeting consisting of 14 symposia, numerous conferences and some 300 scientific posters.  Since much of this research will not be readily available to beekeepers in the U.S., the purpose of this series will be to describe some highlights of the information presented.  The proceedings runs 811 pages; the full edition is available on CD ROM from the IBRA <http://www.ibra.org>

Four presentations comprised the beginning plenary session, in a sense setting the tone for the event.  Two were by those who were honored at the fourth Encontro, Drs. Kerr and Nogueira Neto.  Professor Kerr, who though he is now 82 years old, still works full time at the Federal University of Uberlândia in Minas Gerais, discussed the differences between (Melipona) tropical bees and (Apis) honey bees beyond the presence or absence of a sting apparatus. .  It turns out that Melipona workers more resemble males than queens, and the male bees actually do work in the nest.  Young Melipona virgins leave the nest during swarming instead of the old queen as in Apis, and only mate with one male instead of 10 to 30 normally associated with honey bees.  Finally, Melipona species have from 8 to 20 pairs of chromosomes, whereas Apis has a constant 16.  Dr. Kerr suggested that the variablility in Melipona chromosome number may result from a fracturing or breaking of extant chromosomes.  Most likely they are those involved in determining caste (drone, queen, worker), providing an explanation for the differences between Apis and Melipona bees noted above.

Dr. Nogueira Neto discussed the current status of the culture of Melipona bees from the developing of nests with movable frames to a pump for sucking the honey out of the honey “pots” the bees make.  They do not store honey in combs like Apis bees.  He emphasized that the honey from Melipona is substantially different and commands a higher price than that collected from Apis.  He suggested a new name for this sweet, “iramel,” taken from the indigenous name of the stingless bee (ira) and Portuguese word for honey (mel).  A problem in Brazil is that “iramel” is considered by law to be a native product, which belongs to the state, and thus any trade in the commodity is considered illegal.  He is working with the bureaucracy in Brasilia, Brazil’s modern interior capital, in an attempt to rectify this situation.

Dr. Carminda de Cruz Landim and associates reported on the differences in pheromone-producing glands between stingless (Melipona) and Apis bees.  These are found almost anywhere on the body; suggesting that both types bees can be characterized as a flying “collection of glands.”  The authors presented a chart revealing what is known about the glands present in both kinds of social bees.  These include: Nassanov (on the abdomen), Koschewnikov (inside, near the sting apparatus), tergal (just below the exoskeleton ), tarsal (in the foot), mandibular (around the mouth), Dufour (near the sting apparatus - producing the “alarm” substance) and labial (near the mouth).  Most of these produce substances that are volatile and diffuse into the air, making them difficult to collect and employ in experiments, and this is the reason many have not yet been identified in either kind of bee. 

Dr. Wolf Engels asked “What bee larvae might tell their nurses,” by “taking a look into a superfamily with a lot of kids”.  Communication is necessary, given the highly evolved progressive brood feeding found in honey bees (Apis).  The larvae are monitored continually, and their nutritional needs are determined by workers on each visit.  Sick or parasitized larvae are also detected and eliminated from the colony in the process.  This results from a “larval cry” to “feed me or remove me.” 

A prime example of the “remove me” or “eat me” command is found in a colony’s disposition of diploid drone larvae.  These individuals are produced by inbred Apis queens, which lay many fertilized eggs with two identical alleles at a specific spot, called a “locus,” (the one determining sex) on a chromosome.  Males are normally haploid, but diploid individuals that have two copies of the same sex allele also become males.  They are eaten by workers on the first day of life after hatching from the egg, causing the colony to have to produce a replacement.  Up to fifty percent of the larvae may be affected, weakening a colony through what is called “inbreeding depression.”  The removal behavior does not extend to second- or third-day larvae, but by that time all the diploid drones have been eaten.  Melipona species are not as good as Apis in searching out these larvae, however, and so whereas not one survives in the latter bee species, many appear to do so in the former.

The signal produced by diploid drones is not a “cannibalism substance,” Dr. Engels stated, as originally thought by some investigators.  It is recognition that the skin surface (made up of complicated hydrocarbons) is different from that of normal larvae. 

By contrast, the larval feeding cry is a volatile substance (brood pheromone) put out by the grubs and recognized by workers.  The chemical must last for a period of time, as not all visiting workers have the correct food supply.  Thus, Dr. Engels concluded this “budgeting system” allows workers to feed larvae on demand and only when necessary, “an amazing masterpiece of social evolution.”

In a symposium on Apis biology, Dr. David De Jong, a former U.S. bee inspector, now on the Ribeirão Preto faculty provided an essay on honey bee health.  Location, location, location is as important in bee disease control as in real estate purchase.  Colonies exposed to wet areas, cool conditions and shade are more susceptible to chalkbrood.  Examples may be found in Korea (next to rice paddies) and Brazil (in dense shade).  Colonies that are split, with each resultant unit forced to take care of large brood areas, are also more susceptible (stress) to the chalkbrood fungus (Ascosphaera apis).  Finally, colonies located close together are likely to become infested with pathogens simply by proximity with their neighbors, something rarely occurring in nature. 

Most human management concentrates colonies, which works against the production of disease-resistant bees, according to Dr. De Jong.  So does forcing bees to use wax foundation, moving them to different areas, exposing hives to chemical treatments, and replacing queens.  Requeening is perhaps the most important undertaking of the beekeeper, yet often receives little of the care it deserves in either the rearing of individual queens or introducing them into colonies.  Not only does changing the queen introduce different genes, but it also provides a needed “break in the brood cycle.”  This provides time for affected individuals to be removed, especially known to help in cases of European foulbrood.  In general it can be concluded that many apparent “problems” in bee colonies are provoked by old and failing queens.  Thus, replacing them regularly can often be considered a uniform solution to a myriad of conditions. 

Varroa mite infestation is also increased in colonies that are closer together, as is the potential for other diseases like American foulbrood.  This is because weak colonies become reservoirs for parasites and diseases that then are easily spread to nearby hives by robbing or drifting bees.  Development of susceptible colonies is encouraged by beekeepers using both antibiotics for bacterial control and chemicals to lower mite populations.  One of the lessons of Brazil is that a low population of mites can be tolerated by honey bees indefinitely without treatment.  The Africanized honey bee is an example of how bees being left alone can and have come to terms with challenges in their environment posed by diseases and pests.  There are many resistance mechanisms in healthy bees, but these are all too often ignored and compromised by beekeepers in search of a “quick fix.” 

When looking at how honey bees resist diseases and parasites, one cannot ignore the effect of cell size, Dr. De Jong concluded.  There is some anecdotal evidence that this can play a significant role in colony health, but little formal research to confirm the idea.  Nevertheless, in Brazil the use of smaller cell size foundation (4.7 to 4.9 millimeter cell width) is recommended because it is considered the “normal” size of Africanized honey bee comb.  Most regular foundation has cells 5.2 to 5.4 millimeters in width and often results in higher mite infestation in the colonies.  Dr. De Jong urged beekeepers and researchers not to ignore the significance of cell size in overall colony health. 

Other papers on honey bee pathology revealed the current situation with respect to Varroa mite classification, antibiotic use in bacterial control, and detection and transmission of bee viruses.  Dr. Denis Anderson from Australia provided the latest information on his studies of Varroa classification.  Some 25 different mites have now been identified.  Remarkably, only two (2) have colonized the Western honey bee, Apis mellifera, and become invasive pests.  The one originally found in Brazil is from the Japanese line and considered less virulent.  Unfortunately, the much more aggressive and damaging Korean variety has colonized much of the rest of the Americas, Europe and North Africa.  Currently the Korean variety is also found throughout much of Brazil.

The antibiotic Terramycin® formulated from oxytetracycline hydrochloride for the control of the foulbroods in the Americas, which was effective for some 40 years is now losing much of its therapeutic punch, according to Dr. Mark Feldlaufer and associates at the Beltsville, MD Bee Research Laboratory.  They reported on current efforts to screen colonies for resistance and efforts to make two other products (lincomycin, a macrolide, and tylosin, a lincosaminide).  The Beltsville laboratory also reported new technologies (using molecular biology) to identify viruses in honey bee colonies.  This research shows that viruses can be spread via brood cells, and that in some cases honey bees can harbor four (4) types simultaneously.

Precisely because the honey bee has become such a successful species in Brazil, there is resultant concern about its effect on native pollinators.  This is true throughout the world, as more and more species are purposefully or inadvertently introduced into ecosystems they are not native to.  In 1998, a Brazilian workshop attended by 61 scientists from 15 countries on the conservation and sustainable use of pollinators in agriculture with an emphasis on bees resulted in the “São Paulo Declaration on Pollinators.”  This model has now been used in a great many countries that have also produced pollinator initiatives.  It consists of:

1.      Monitoring pollinator decline and its effect on commercial agriculture.

2.      Addressing the lack of classification information on native pollinators.

3.      Promoting conservation, restoration and sustainable use of native pollinators.

The Brazilian Pollinators Initiative (BPI) has transformed into and International Pollinators Initiative (IPI) and has a World Wide Web presence <http://www.webbee.org.br>.  Other papers in the symposium on pollinator initiatives included monitoring the spread of non-native bumble bees (Bombus ruderatus) in southern South America, and cataloging floral resources needed by tropical stingless bees and carpenter bees.

A related symposium discussed the incredible bee diversity in the Brazilian tropics.  One study examined the tree canopy in Amazonia.  This involved using towers 30 to 45 meters (in excess of 100 feet) in height to collect representative species.  Another described the number of bees (1994 individuals representing 24 species or groups) in the family Euglossidae (orchid bees) captured in the Atlantic eucalyptus forest along Brazil’s coast.  The authors concluded that the number of these bees was a good measurement of  the extent of habitat destruction in one of the last remaining mature (climax) forests found in the country.  Another paper examined the historical evolutionary development of all native bees in South America.  Evidence is provided that some originated in the Cretacious period, when South America was still connected to Africa as part of a land mass known as Gondwanaland, whereas others appear to be the result of much more recent arrivals from North America.

 

6^th Brazilian Encontro and 8^th International Conference on Tropical Bees Part II

 

By

 

Malcolm T. Sanford

http://apis.shorturl.com

 

In my first article on the sixth Brazilian Encontro, I closed with the following statement: “Evidence is provided that some (native bees) originated in the Cretacious period, when South America was still connected to Africa as part of a land mass known as Gondwanaland, whereas others appear to be the result of much more recent arrivals from North America.”

The most studied and sensational recent arrival to Brazil of course was that of the African honey bee in 1957.  The story is well known and was detailed in a previous series of mine in this journal.  A symposium on the Africanization process at the 6^th Encontro provides some current insight about the present status of this controversial insect in the Americas.  Dr. Walter (Steve) Sheppard from the University of Washington said in an overview paper that the ability of researchers to detect subspecific genetic ancestry has increased such that controversial, but not mutually exclusive, views of the Africanization process have emerged.  The literature shows that both natural and artificial (beekeeper) selection have influenced the process.  It is not surprising that in areas where tropically-adapted bees have a selective advantage, there is the lowest genetic influence of European bees.  In areas where no clear advantage exists there is in fact a mixed genetic composition resulting from both beekeeper selection for European traits and gene flow from resident non-Africanzed feral populations.

Dr Marco Antonio Del Lama provided some historical information on what researchers believe are the major evolutionary lines of honey bees.  These insects appear to have colonized both Europe and Africa resulting in three distinct branches: 1) South or central African (A); 2) North African and West European (M); and 3) North Mediterranean (C).  Two other lineages have been proposed to include both Near and Middle Eastern species. 

Introductions to the New World included many of these branches, but most striking is that of Africanized honey bees, according to Dr. Del Lama.  He used both molecular (enzyme analysis) and measurements (morphometrics) to characterize populations in South America, which he concludes vary greatly depending on region.  In Uruguay, he said that a hybridization process has been “completed.”  Mostly Africanized bees are found in the north and a hybridized zone between Africanized and European bees exists in the south.  In Chile, no Africanized populations could yet be discerned, indicating the country is populated by European lines only at the present time. 

Colombia is a different story.  The bees are much more like the Brazilian Africanized population.  The data reveals, therefore, that Africanized bees are able to “explore the different environments” found in the country.  In general, Dr. Del Lama’s data agree with Dr. Sheppard’s conclusions.  He also found that Africanized honey bee populations are “very representative of the genetic variation of African bees from the Sub-Saharan area.”  Soon he hopes to determine the number of both queens and drones that made up the introduced or “founding” population of African bees.  The conclusion so far is that only twenty-six (26) queens were responsible for this biological revolution.

The Africanized honey bee in Mexico was discussed by Dr. Deborah Smith.  She examined populations in the south (Yucatan) and the north (Linares, Nuevo León).  In the former area European populations were high, whereas in the latter they were low prior to arrival of the Africanized bee.  The basis of this research was to examine shifts in the mitochondrial DNA (mtDNA).  It is known that this is only inherited through the mother, and most areas colonized by Africanized honey bees undergo a change from European to African mtDNA.

During three (3) years (1990-1993), 1738 swarms were captured in the Linares area.  In the first year no European mtDNA could be found, but over the next it became predominant in populations.  However, the European type could still be found, especially during times where reproductive swarming was high.  Many of these it is thought came from managed rather than feral colonies.  When reproductive conditions were limited and migratory swarming predominated, the proportion of Africanized mtDNA increased.  Colonies with African mtDNA had smaller average cell sizes than Europeans, and this appears to be the case in places where Africanized bees have colonized in the United States. 

Yucatan is unique in that it had one of the most dense populations of European honey bees prior to arrival of the Africanized honey bee.  The bee type was considered mostly east European.  By 1998 (12-13 years after arrival) samples collected in 312-331 managed colonies and 31 feral nests showed an almost complete in mtDNA from European toward African.   Most extreme was the shift in feral colonies and 219 managed colonies had profiles that resembled Venezuelan Africanized bees.

Although the methods were different in comparing populations in both areas of Mexico, they revealed similar patterns.  The fact that many genetic profiles resembled those of bees from Venezuela reveals that perhaps feral African or Neotropical bees acquired a unique genetic composition early on.  This consists of a “high frequenty of African-typical nuclear alleles, and a low frequency of East and West European alleles, on a cytoplasmic background that is predominantly African with as small West European contribution."

Dr. Robert Danka says that the shift in genetic composition reported by others should not be surprising, given the “remarkable reproductive advantage accruing to Africanized honey bee (AHB) magnified by the rather poor ability of European bees (EHB) to survive in the tropics, especially the lowlands.”  High mortality of managed EHB colonies (up to 15 percent in many cases) is reported, most often linked to predation and unpredictable food shortages, common in tropical regions, but not in temperate areas.  The vigorous nest defense mechanisms and high rates of absconding (migratory swarming) in AHB are thought to be effective responses to both problems. 

The mechanisms that favor continuity of the African genome in the Americas are complex.  Drs. S. Schneider and G. Di-Grandi Hoffman conclude: “No single factor determines the ability of African bees to displace European bees.  The continuity of the African genome in the Americas arises from a complex interaction of population dynamics, genetic phenomena, and physiological and behavioral mechanisms.”  These include 1) African-patriline advantages during queen replacement, 2) differential use of African and European sperm by queens, 3) nest usurpation by African swarms and 4) decreased developmental stability in hybrid workers.

There was no advantage seen for AHB drones during the queen rearing phase, but plenty of evidence suggests there was subsequently.  Virgin queens with AHB fathers emerged sooner, developed faster and were fiercer fighters during queen rivalry sessions. Queens inseminated by equal numbers of AHB and EHB drones preferentially used AHB sperm, according to experimentation, and thus Africanization of managed European bees "could act as a barrier to the introgression of European paternal alleles into African colonies." 

Nest usurpation (takeover of colonies) by AHB is generally considered a given in most areas where this insect has been introduced.  However, the condition of the colony (weak colonies are more susceptible) and timing are critical as AHB has both a reproductive (swarming) and absconding (migrating) season or cycle to take into consideration.  The study concludes: "re-queening colonies during certain times of the year could actually increase the vulnerability to becoming  African...Identifying when swarming  and absconding by feral African colonies occurs (sic) in a given area and  avoiding re-queening at those times could be critical to maintaining European matrilines (queens)."

Finally, there is a major controversy about the Africanization process surrounding what is called  “hybrid inferiority.”  The idea is that if hybrid workers are at a disadvantage, this could be a big reason that European bees decline over time.  Using four genotypes (AA, AE, EE, EA, representing A and E queens mated to both kinds of drones respectively), the authors found that what they call “developmental stability” was in fact higher in pure Africans (AA) than the other genotypes, providing more evidence for this phenomenon.

Dr. Kirk Visscher discussed the history and current status of Africanized honey bees in the United States.  The insects first were reported in 1990 in Texas, followed by Arizona (1993), New Mexico (1994), Nevada (1998) and California (1984).  The latter figure I believe is a typographical error in the published paper in the proceedings and probably should be 1994.  However, the date closely coincide with a first report of Africanized bees in Lost Hills, California in a load of pipe off a ship in 1985; these were, however, exterminated at the time and cannot be said to contribute to California’s current population.

The bottom line is that the bees have continued to spread northward in the West, and in California have “sharply expanded” their range in California, including all the southern counties of the state.  Looking to the east, one of the greatest puzzles is why the invasion fizzled in eastern Texas and did not spread into neighboring states (Louisiana, Mississippi, Alabama, and Florida).  The good news is that in the fourteen years they have been in the United States, Africanized bees have caused fewer stinging incidents and deaths than initially expected.  This figure even  represents less than that projected for a resident population of European bees.  Dr. Visscher suggests that this is due to publicity, education and better prepared emergency agencies.

The invasion, nevertheless, continues and the Africanized honey remains a threat to populations, especially in southern Arizona where there have been numerous incidents and especially animal deaths.  In most areas, after first detection, the Africanized population increases slowly and then rapidly as seen in data from Imperial County, California. 

Response to population increase has come in several ways.  Prevention of  problems by “bee proofing” structures, educating the populace and diluting the gene pool has been of limited help.  The first strategy is costly and often not considered necessary by much of  the general public unless a stinging incident occurs.  The last has also not been as successful as first thought.  Increased fear of bees has cost beekeepers locations as happened in most of Latin America, reducing bee populations in general.  Frequent re-queening is costly, and neglect may result in extremely defensive hybrids.

Interception refers to situations where bees are collected before they move into structures.  A dilemma, according to Dr. Visscher, is that initially swarms are not defensive and can be captured and destroyed easily.  However, if one waits too long and the colony becomes established the situation can worsen.  Professional removal services are expensive.  Some entrepreneurs have instigating trapping procedures, however, “this works best with coordination over a reasonably large area, so it has mostly been restricted to golf courses and other large land holdings.”  Perhaps the best example of this is the program developed in Ribeirâo Preto itself where the Encontro took place.  The place was invaded by large numbers of Africanized honey bee swarms, which were attracted to the oozing sugar cane stubs left after the harvest.  A trapping program was necessary to intercept the swarms before they could take up residence in the area.

Like so many phenomena in agriculture, treatment should be a last resort, but often isn’t.   It involves safe removal of bees from structures and other locations that are difficult to find and access.  Again, private enterprise has taken the lead, but increased stinging incidents may bring on enough complaints in certain areas that agencies and local governments will have to provide these services.  Dr. Visscher says that it is likely that problems with these bees will continue to increase, especially in newly colonized areas of California.  The recent rush to rent colonies of honey bees to pollinate almonds in the Bear Republic may in fact be aiding the process as noted by Chuck Norton in his article in the May 2005 American Bee Journal.

Dr. Visscher concludes: “Although climatic factors are surely important, the final distribution of Africanized bees may be determined by competition with European honey bees as much as by climatic barriers.  The abundance of European bees, in turn is dependent on beekeeping and on response of unmanaged honey bee populations to parasites and diseases.  Africanized bees arrived in the US simultaneously with the spread of parasitic mites, especially Varroa, which sharply reduced feral populations.  There is considerable, mostly anecdotal, indication that bee populations are recovering, even in areas not colonized by AHB, and this will likely influence the eventual spread of Africanized bees.”

 

References:

 

1.      M.T. Sanford, Apis Newsletter, IFAS University of Florida , July and August 1989<http://apis.ifas.ufl.edu/apis89/apjul89.htm#1> and<http://apis.ifas.ufl.edu/apis89/apaug89.htm#1>, accessed July 19, 2004.

2.      Hotel JP Home Page <http://www.hoteljp.com.br/>, accessed July 19, 2004.

3.      Sixth Encontro de Abelhas Home Page <http://rge.fmrp.usp.br/abelhudo/>, accessed July 19, 2004.

4.      International Bee Research Association Home Page <http://www.ibra.org.uk/>, accessed July 19, 2004.

5.      For more information, e-mail <aronisattler@yahoo.com.br>.

6.      Page <http://www.apacame.org.br>, accessed July 19, 2004.

 

6^th Brazilian Encontro and 8^th International Conference on Tropical Bees Part III

 

By

 

Malcolm T. Sanford

http://apis.shorturl.com

 

In contrast to many scientific meetings about honey bees, the sixth Brazilian Encontro also included a symposium on honey bee products as they relate to human health.  This is an area that is often ignored by those practicing modern medicine, however, there is growing scientific evidence that this should no longer be the case.  According to Dr. Shona Blair of the University of Sydney, Australia’s School of Molecular and Microbial Sciences, the healing properties of honey come in good measure from its antimicrobial activity. She concludes, “The antimicrobial activity of honey has been reported to range between minimum inhibitory concentrations (MIC) of 2% to over 50%, depending on the microorganism against which it was tested and the floral source of the honey.”

One factor involved in honey’s antibacterial activity is high sugar concentration, resulting in “low water activity,” according to Dr. Blair, often below that required by most bacteria.  This is true of most concentrated sugar solutions, however, honey retains its properties after being diluted and has been found to be “significantly more effective than sugar solutions of similar water activity.”  Another factor is that glucose oxidase, secreted from the pharyngeal glands of the honey bee, reacts with glucose to form hydrogen peroxide at concentrations high enough to inhibit bacterial growth. 

Some of honey’s healing attributes go beyond high sugar content and hydrogen peroxide formation, and appear to depend on floral source, Dr. Blair writes:  The best known are Leptospermum scoparium (manuka) from New Zealand and certain other Leptospermum (jelly bush) honeys from Australia.  The active components of these have yet to be identified, but their effects on Escherichia coli, Staphylococcus aureus and Acinetobacter spp. are encouraging, especially given that many bacteria are already or are quickly becoming resistant to antibiotics.  Honey also has therapeutic properties on human tissues, which also contributes to healing.  It stimulates human cells grown in vitro (Petri dish) and activates macrophages (scavenger cells), stimulating them to release a number of cytokines, such as interleukin 1-alpha and 6. 

Given all the evidence, Dr. Blair concludes: “Honey is a grossly underutilized wound dressing.”  This is also true for chronic ulcers that do not respond to conventional treatment, while honey has given remarkable results under complex circumstances, even involving antibiotic-resistant organisms.

Dr. Rose A. Cooper of the University of Wales reports that honey was routinely used in United Kingdom (UK) hospitals for treating wounds from the Middle Ages until the 1950s, when the practice was almost entirely discontinued due to the rapid rise of antibiotics.  In the 1970s and  80s, new sorts  of  bandages emerged, all but eliminating those made from gauze or lint.  However, there quickly emerged antibiotic-resistant strains of microorganisms due to the use and “misuse” of antibiotics, and more ominously, “selection of strains with multiple antibiotic resistance.” 

Dr. Cooper provided historical information on the curative aspects of honey and a listing of specific clinical evidence.  The mindset of many doctors, and more importantly the regulators, is beginning to change with reference to honey use, she says. A honey-impregnated wound dressing was approved in March, 2004, and more products are expected to follow.  Most “medical practice has adopted an evidence-based approach, and practioners will only modify procedures when provided with suitable evidence,” Dr. Cooper concludes.  Fortunately, this is gradually accumulating and Dr. Cooper is confident that opinion is rapidly changing concerning the use of honey in treating wounds.

The use of propolis has been on the increase in Brazil ever since Dr. Matsuno from Japan revealed anti-tumor activity from Brazilian propolis, triggering a “boom,” according to Drs. Mitsuo Matsuko and Jun Nakamura.  This followed the Japanese rise in propolis use in conjunction with the 30^th  Apimondia Congress, in Nagoya, Japan in 1985.  All this attracted many basic and clinical researchers. 

The authors provide the following chronicle of events:

1973  Propolis importation to Japan reported  in the journal Bee World.
1979  Research on propolis began at Tamagawa University
1985  Apimondia Congress, Nagoya, the beginning of widespread propolis use in Japan
1987  Japanese Propolis Conferences (JPC) begin
1991  Dr. Matsuo reports anti-tumor properties for three compounds in propolis
1997  Propolis Researcher’s Association (PRA) begins
1998  Japanese Society of  Alternative Medicine started, name changed  to Japanese  Society of  Complementary Alternative Medicine (JSCAM) in 2000.

Since 1997, the PRA has had a litany of speakers from Eastern Europe, USA, Uruguay, Japan, and of course Brazil.  All this activity has been beneficial, the authors write, but also has resulted in misconceptions regarding different kinds of propolis.  That from the European poplar, for example, is quite different that that from Baccharis (Brazilian type).  The former is rich in flavonoids, while the latter type has more diterpenes.  The flavonoid quercetin has been incorporated into a propolis standard by many marketers, when in fact the propolis used in products often has little or none.  In addition, the authors state that many Japanese believe propolis comes from Eucalyptus, simply because dealers have made that claim.

The PRA was established precisely because of the confusion, the authors say.  It has encouraged research on one hand while on the other it has provided education for both consumers and health practitioners.  The bottom line is that both should always consider the kind of propolis being used.  Papers presented at the PRA run the gamut from basic properties to manufactured products, including specifically those concentrating on:

Pharmaceutical/medical/biochemical properties of propolis
Active ingredients found in propolis
In vivo (live) and in vitro (Petri dish) activity of propolis
Clinical trials and safety tests
Food engineering aspects
Standards and grading of propolis products

Scientific papers have appeared in a variety of scientific journals, including Biological Pharmaceuticals Bulletin (propolis effect on human cells lines and leukemia), Food Chemistry (antioxidant properties of propolis), Org. of Biological Chemistry, and Cancer Letters.

The complexity of Brazilian propolis is evident when considering a paper by Dr. Yong Park, from the university of Campinas, on its biodiversity.  It is evident that the properties of propolis are almost wholly dependent on plant ecology, Dr. Park says, when he analyzed 500 samples collected by Apis mellifera in several regions of Brazil.  These were put into 12 groups (5 from southern, 6 from northeastern, and one from southeastern and western areas).  These were extracted in 80% ethanol and then compared for a number of properties, including antioxidant activity, antimicrobial activity, anti-inflammatory activity, cytotoxicity (cell poisoning), and anti-HIV activity.  Finally, other biologically active substances were also isolated and identified.

As expected, Dr. Park found a great variety of activity in the propolis samples.  He meticulously recorded his results in various charts and tables, and concludes that all this information punctuates the complexity of studying propolis and its biological activity. 

Dr. Walter Ferro Morales from Uruguay (who unfortunately only sent a paper and could not attend) wrote on the antioxidant and healing properties of Uruguayan propolis.  This included a case study of 229 patients reported at the 10th Latin American Surgical Congress.  Gauze impregnated with 2 percent propolis solution was found to be a superior treatment to conventional applications.  Unfortunately, Dr. Morales did not take a page out of Dr. Park’s book and so readers are left with an incomplete story as to the kind of propolis used in the Uruguayan study.

Although there are standards for both propolis and honey in Brazil, the same is not true for honey mixed with propolis.   Alexandre Bera and colleagues state that consumers would better accept the latter product, because alcoholic extracts of propolis have “a strong taste and may cause rejection.”  They report using the same methods for standardizing honey and propolis separately, using guidelines from the Codex Alimentarius.  They concluded that the method for moisture (18.67%) and ash (0.003%) determination were suitable for all samples.  The same was true for reducing sugars (71.88%), apparent sucrose (3.47%), hydroxmethylfurfural or HMF (44.03%) and insoluble solids (0.0007%).  Determination for diastase on the other hand had to be adapted specifically for the honey and propolis mixture.  This study suggests that it would be relatively easy for those who wish to develop and market a product based on a mixture of honey and propolis to develop an adequate standard in the market place.

Dr. G.J. Padovan and colleagues compared the quality of honey being marketed in the Ribeirão Preto region of the state of São Paulo, Brazil with reference to the testing mechanisms in place.  The authorities use physiochemical tests, which target the normal falsifications found in honey.  However, the investigators showed that these classic procedures are not as sensitive as the carbon isotope ratio (C12/C13) determinations with a mass spectrograph, which detected more adulterated samples (11 of 20 vs. 5 of 20).  They conclude that more adulteration is going on than is currently detected by official means.  Thus, the techniques used to adulterate honey are technologically superior to the “currently legislated testing mechanisms.” This information is being presented to the government to encourage more rigor.

Several other areas of interest were explored through symposia at the Encontro.  These included sustainable development in beekeeping, sensory perception and learning in honey bees, stingless bee biology and advances in genomic and molecular biology.  Several papers on sustainability emphasized the wide range of possibilities the keeping of both Apis and non-Apis bees has in the developing world.

Dr. Wyatt Mangum reported a number of innovations he has found around the world.  For example, one Bolivian beekeeper rents out his expertise in managing colonies of highly defensive Africanized bees owned by others.  He also described innovations in keeping non-Apis bees by beekeepers who have designed nesting materials and other technologies to harvest colonies from the wild.  Unfortunately, all these activities, Dr. Mangum concluded, are threatened by the pervasive worldwide movement of both honey bees and bee pests out of their native ranges, including diseases  like American foulbrood (Paenibacillus larvae larvae), Tropilaelaps clarae (an Asian parasitic mite), Apis  mellifera scutellata (overly defensive Africanized bees), Apis  mellifera capensis (honey bees producing “false queens”), and Aethina tumida (small hive beetle).

As would be expected, a variety of papers were also presented in the bee pathology area.  Several of the most relevant included original research on Varroa mite infestation.   One Venezuelan study revealed that “survivor” colonies of Africanized honey bees that had never been treated had a greater number of non-reproducing female mites than would be expected.  A bioassay has been developed to reveal how Varroa mites are attracted to comb and brood.  It was concluded: “With this system, the mites made choices similar to those they make in honey bee colonies, such as preferring old brood comb wax to newly constructed comb and drone brood food over royal jelly.”  The former conclusion is being examined as a possible natural control of Varroa by incorporating new and renovating aged comb extracts to wax foundation.  Another study concluded that Varroa infestation in worker brood decreased as the amount of brood in a colony increased, however, the relative infestation rate in drone brood remained relatively constant.  An apparatus was described to film activities inside a colony, producing the first “real time” images of interactions between worker bees and Varroa mites.

No conference on bees is complete these days without some treatment of diseases.  American foulbrood is so far not recorded in Brazil.  However, beekeepers and researchers are concerned about its introduction.  A study reported at the Encontro revealed the relative amount of resistance to antibiotics that spores of the causative organism, Paenibacillus larvae larvae, have.  Honey imported from five countries showed that 13 out of 310 AFB bacteria samples were resistant to tetracycline (Terramycin®), but 227 were resistant to the sulfa drug sulfamethazine, 198 to sulfathiazole, and 187 or sixty percent of samples were resistant to both sulfas.  Finally, only 11 or 3.5 percent of samples were resistant to all three kinds or antibiotics.  The practical implications of this are not good.  Not only is it likely that the disease will be introduced to Brazil, but when it is, there is a good chance it will be resistant to one or more of the treatments currently in use around the world.

As I said in the initial article on this series, the magnitude of the published document produced containing the proceedings of the 6^th Brazilian Encontro and 8^th International Conference on Tropical Bees defies giving it justice in a few short articles.  It is my fervent hope, however, that the readership will get a flavor of the meeting from these three summary articles.  It might even be that some are stimulated enough to buy the proceedings on CD ROM from the International Bee Research Association <http://www.ibra.org>.  The information certainly deserves wider dissemination in the apicultural community. 

 

Acknowledgements:

 

Thanks to Dr. Lionel Gonçalves for encouraging me to write this series and providing me with a copy of the proceedings.   I would like to specifically acknowledge the help of Dr. David De Jong, who conscientiously read and edited each article before it went to press.