206 Recent Patents on Anti-Infective Drug Discovery, 2009, 4, 206-213

Rediscovering the Antibiotics of the Hive

Laïd Boukraâ* and Siti A. Sulaiman

Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang
Kerian, Kelantan, Malaysia

Received: May 30, 2009; Accepted: July 9, 2009; Revised: July 18, 2009
Abstract: Honey and other bee products were subjected to laboratory and clinical investigations during the past few
decades and the most remarkable discovery was their antibacterial activity. Honey has been used since ancient times for
the treatment of some diseases and for the healing of wounds but its use as an anti-infective agent was superseded by
modern dressings and antibiotic therapy. However, the emergence of antibiotic resistant strains of bacteria has confounded
the current use of antibiotic therapy leading to the re-examination of former remedies. Honey, propolis, royal jelly and bee
venom have a strong antibacterial activity. Even antibiotic-resistant strains such as epidemic strains of methicillin-
resistant Staphylococcus aureus (MRSA) and Vancomycine resistant Enterococcus (VRE) have been found to be as
sensitive to honey as the antibiotic-sensitive strains of the same species. Sensitivity of bacteria to bee products varies
considerably within the product and the varieties of the same product. Botanical origin plays a major role in its
antibacterial activity. Propolis has been found to have the strongest action against bacteria. This is probably due to its
richness in flavonoids. The most challenging problems of using hive products for medical purposes are dosage and safety.
Honey and royal jelly produced as a food often are not well filtered, and may contain various particles. Processed for use
in wound care, they are passed through fine filters which remove most of the pollen and other impurities to prevent
allergies. Also, although honey does not allow vegetative bacteria to survive, it does contain viable spores, including
clostridia. With the increased availability of licensed medical stuffs containing bee products, clinical use is expected to
increase and further evidence will become available. Their use in professional care centres should be limited to those
which are safe and with certified antibacterial activities. The present article is a short review of recent patents on
antibiotics of hives.
Keywords: Bee product, honey, royal jelly, propolis, bee venom, antibacterial, antibiotic resistance.

INTRODUCTION
Natural products have been used for thousands of years
in folk medicine for several purposes. Among them, propolis
has attracted increased interest in recent years due to its
antimicrobial activity against a wide range of pathogenic
microorganisms. Various studies attribute antibacterial, anti-
fungal, anti-inflammatory, antiproliferative and anticancer
potentiating properties to honey [1]. Honey, propolis, and
royal jelly, products originating in the bee hive, are attractive
ingredients for healthy foods. Several aspects of this use
indicate that honey also has functions such as antibacterial
[2], antioxidant [3] and anti-inflamatory [4]. Propolis is a
resinous substance produced by honeybees. This substance
has been used in folk medicine since ancient times, due to its
many biological properties to possess, such as antioxidant,
antimicrobial, anti-inflammatory, and immunomodulatory
effects, among others [5]. Given the exceptional biological
properties attributed to it, royal jelly (RJ) has considerable
commercial appeal and is today utilised in many sectors,
ranging from the pharmaceutical and food industries to the
cosmetic and manufacturing sectors [6]. Royal jelly has been
demonstrated to possess numerous functional properties such
as antibacterial and anti-inflammatory activity [7, 8]. Biolo-
*Address correspondence to this author at the Department of Pharmacology,
School of Medical Sciences, Universiti Sains Malaysia, Health Campus,
16150 Kubang Kerian, Kelantan, Malaysia; Tel: +609-7664707;
Fax: +609-7653370; E-mail: lboukraa@kk.usm.my
gical activities of honey, propolis, and royal jelly are mainly
attributed to the phenolic compounds such as flavonoids.
Recent studies have shown that bee venom exhibits anti-
bacterial activity [9, 10]. The rapid emergence of antibiotic-
resistant pathogens poses increasingly serious health
concerns worldwide. Approximately 70% of bacteria that
cause infections in hospitals are resistant to at least one of
the antibiotics most commonly used to treat infections. This
antibiotic resistance is driving up health care costs,
increasing the severity of disease, and the fatality of certain
infections. The annual cost of treating antibiotic-resistant
infections in the US is estimated to be as high as $30 billion.
Sepsis is another serious medical condition resulting from
severe inflammatory response to systemic bacterial
infections [11]. Antimicrobial peptides (also known as
natural antibiotics) comprise a large group of molecules that
are capable of killing a broad spectrum of pathogens with
similar activities against both antibiotic-susceptible and -
resistant bacterial strains and extremely low risks of
developing resistance [12]. More desirably, many have
capacity to bind bacterial endotoxin and neutralize bacte-
rium-induced inflammatory response. Because of the dual
capability to kill bacteria and neutralize endotoxins, these
antimicrobial peptides hold great promise as a new class of
antimicrobial and anti-sepsis agents [13]. As natural products
garner attention in the medical field, the emergence of
antibiotic-resistant strains of bacteria has confounded the
current use of antibiotic therapy, leading to the re-
examination of earlier remedies such as honey, royal jelly,

1574-891X/09 $100.00+.00 © 2009 Bentham Science Publishers Ltd.

Antibacterial Bee Products Recent Patents on Anti-Infective Drug Discovery, 2009, Vol. 4, No. 3 207

propolis and bee venom. The rediscovering of bee products Table 2. Average Composition of Honeys and Ranges of Values
as antimicrobials is discussed within this mini-review. (White, 15)

HONEY Component Average Standard Range

Honey is essentially a highly concentrated water solution (% Except pH and Deviation
of two sugars, dextrose and levulose, with small amounts of Diastase Value)
at least 22 other more complex sugars. Many other Water < 17.2 1.5 13.4 - 22.9
substances also occur in honey, but the sugars are by far the
major components (Table 2). The principal physical Fructose 38.2 2.1 27.2 - 44.3
characteristics and behaviour of honey are due to its sugars,
Glucose 31.3 3.0 22.0 - 40.7
but the minor constituents (such as flavouring materials,
pigments, acids, and minerals) are largely responsible for the Sucrose 1.3 0.9 0.2 - 7.6
differences among individual honey types [14]. Honey, as it
is found in the hive, is a truly remarkable material, Maltose (reducing 7.3 2.1 2.7 - 16.0
elaborated by bees with floral nectar, and less often with disaccharides calculated as
honeydew. Nectar is a thin, easily spoiled sweet liquid that is maltose)
changed by the honey bee to a stable, high-density, high- Higher sugars 1.5 1.0 0.1 - 8.5
energy food [15]. Topical application of honey to burn
wounds and other wounds has been found to be effective in Free acids (as gluconic 0.43 0.16 0.13 - 092
controlling infection and producing a clean granulating bed acid)
[16-19]. Many studies [20,21] have shown that honey has Lactone (as glucolactone) 0.14 0.07 0.0 - 0.37
antibacterial activity in vitro, and clinical case studies have
shown that application of honey to severely infected Total acid (as gluconic 0.57 0.20 0.17 - 1.17
cutaneous wounds is capable of clearing infection from the acid)
wound and improving healing.
Ash 0.169 0.15 0.020 - 1.028
Table 1. Approximate Equilibrium between Relative Humidity
(RH) of Ambient Air and Water Content of a Clover Nitrogen 0.041 0.026 0.000 - 0.133
Honey (White, 15) pH 3.91 - 3.42 - 6.10

Diastase value 20.8 9.8 2.1 - 61.2

Air (%RH) Honey (% Water Content)

50 15.9
55 16.8
60 18.3
65 20.9
70 24.2
75 28.3
80 33.1
Honey has an osmolarity sufficient to inhibit microbial
growth [22] Table 1, but when used as a wound contact
layer, dilution by wound exudate reduces the osmolarity to a
level that ceases to control infection [22, 23]. The fact that
the antibacterial properties of honey are increased when
diluted was clearly observed and reported in 1919 [24]. The
explanation for this apparent paradox came from the finding
that honey contains an enzyme that produces hydrogen
peroxide when diluted [25]. This agent was referred to as
'inhibine' prior to its identification as hydrogen peroxide
[26]. Hydrogen peroxide is a well-known antimicrobial
agent, initially hailed for its antibacterial and cleansing
properties when it was first introduced into clinical practice
[27]. In more recent times it has lost favour because of
inflammation and damage to tissue [28-30]. However, the
hydrogen peroxide concentration produced in honey
activated by dilution is typically about 1000 times less than
in the 3% solution commonly used as an antiseptic [31].
Although the level of hydrogen peroxide in honey is very
low it is still effective as an antimicrobial agent. It has been
reported that hydrogen peroxide is more effective when
supplied by continuous generation with glucose oxidase than
when added in isolation [32]. Besides the hydrogen peroxide
activity, additional non-peroxide antibacterial factors have
been identified [33, 34]. Manuka (Leptospermum scoparium)
honey from New Zealand has been found to have substantial
levels of non-peroxide antibacterial activity. This is
associated with an unidentified phytochemical component.
Antibiotic-resistant strains have also been studied and
found to be as sensitive to honey as the antibiotic-sensitive
strains of the same species [35] (Table 3). In patients with
wounds infected with antibiotic-resistant strains of bacteria,
not responding to antibiotic therapy, good results have been
achieved after five weeks of application of honey [36].
Wounds infected with MRSA have also been cleared of
infection and healed by application of honey including a leg
ulcer [21], cavity wounds resulting from haematomas [37]
and surgical wounds [38]. Also vancomycin-resistant
Enterococci (VRE) have been reported to be sensitive to
honey [39, 40]. The microorganisms causing nosocomial
infections are known to be resistant to more than one
antibiotic [41]. Honey seems to be a good alternative for
managing such infections. In fact, the most isolated bacteria
from burns and wounds, namely Staphylococcus aureus and
Pseudomonas aeruginosa, have been found to be sensitive to
208 Recent Patents on Anti-Infective Drug Discovery, 2009, Vol. 4, No. 3 Boukraa and Sulaiman

honey action [42, 43]. In some recent studies and inventions Table 3. Infection Caused by Bacterial Pathogens that are
honey is mixed to other natural products for best potency. Sensitive to the Antibacterial Activity of Honey
Honey mixed to Aloe vera which provides a topical (Molan, 31)
formulation that helps heal damaged skin while providing
soothing relief. It also contains anti-bacterial, anti- Infection Pathogen
inflammatory and anti-fungal agents [44]. Another patent
invention proposes a creamed formulation of honey; the Anthrax Bacillus anthracis
composition may include a thickening agent such as an
Diphteria Corynebacterium
alginate-based material [45]. A recent patent proposes an
dithteriae
embodiment of infected area containing honey with the
essential oils of an herb from the family Lamiaceae [46]. A Diarrhea, septicemia, urinary infections Escherichia coli
recent study compared healing of abscess wounds with either wound infections
honey or chlorinated lime and boric acid solution (EUSOL),
a standard of care in many parts of the world. Honey-treated Ear infections, meningitis, respiratory Haemophilus influenzae
wounds demonstrated quicker healing and a significantly infections, sinusitis
shorter hospital stay compared to those treated with EUSOL. Pneumonia Klebsiella pneumoniae
The investigators concluded that honey is a superior wound
dressing to EUSOL and is recommended for dressing Meningitis Listeria monocytogenes
infected wounds [47]. Tuberculosis Mycobacterium
There have been many studies identifying properties of tuberculosis
the non-peroxide activity and attempts have also been made
Infected animal bites Pasteurella multocida
to identify the fraction responsible for this additional activity
have also been made but have so far been unsuccessful. Septicemia, urinary infections, wound Proteus species
Honey yielded from the Manuka tree (Fig. 1) has a strong infections
flavour with an herby, woody characteristic and is dark in
colouring. Manuka honey contains high concentrations of Urinary infections, wound infections Pseudomonas aeruginosa
aromatic acids with the dominant being 2-hydroxy-3- Diarrhea Salmonella species
phenylpropionic acid [48, 49] and the aromatic acids are in a
Septicemia Salmonella cholerae-suis

Typhoid Salmonella typhi

Wound infections Salmonella typhimurium

Septicemia, wound infections Serratia marcescens

Dysentery Shigella species

Abscesses, boils, carbuncles, impetigo Staphylococcus aureus

wound infections

Urinary infections Streptococcus faecalis

Dental caries Streptococcus mutans

Ear infections, meningitis, pneumonia Streptococcus pneumoniae

sinusitis

Ear infections, impetigo, puerperal fever Streptococcus pyogenes

rheumatic fever, scarlet fever, sore
throat, wound infections

Cholera Vibrio cholerae

much greater concentration in Manuka than New Zealand

clover honey (1000 times greater) [50]. Honey is a rather
runny substance, with the result that its use in a wound
dressing is messy and impractical and the wound dressing
has no absorbing capacity, so that on an exuding wound it
rapidly becomes unevenly liquefied and runs off the wound.
A recent invention proposes a wound dressing which
comprises a wound-contacting layer composed of a mixture
Fig. (1). Leptospermum scoparium: the botanical origin of Manuka of honey and a moisture-absorbing agent; a water-permeable
Honey fabric backing layer; and an intermediate layer comprising
Source: www.NZPlantPics.com 2009 water-permeable fabric impregnated with a mixture of honey
Antibacterial Bee Products
and a moisture-absorbing agent [51]. Synergistic effect has
been noticed between honey and natural products such as
royal jelly [52, 53] and starch [42, 43].
ROYAL JELLY
Raw royal jelly (RJ) is produced by honey bees in a
similar manner to honey, but the composition thereof is
fundamentally different. This raw RJ is the foodstuff for
queen bees that young worker bees have synthesized and
manufactured within their bodies, using honey and flower
pollen as raw materials. More specifically, worker bees
collect flower pollen, they knead this pollen together with
honey from the flowers into a “dumpling” form that they
carry back to the nest, these “dumplings” are fed to young
bees for up to ten days after birth, then after passing through
the digestive systems of these bees, the raw RJ is recons-
tituted by the salivary glands and is secreted as an
“astringent-tasting milky-white liquid” from endocrine
glands in the heads of the bees [54-56] Fig. (2).
A B
Fig. (2). Queen Larva floating in Royal Jelly (FAO, 60).
A. A 3-day old queen larva floating in royal jelly. The cell is almost
ready for harvesting.
B. A 5-day old queen larva in a newly sealed cell just before
pupation. Not much royal jelly is left.
Raw RJ contains a moderate amount of the three main
nutritional components (proteins, sugars, and fats) Table 4; is
rich in vitamins, minerals, and amino acids; and further-more
contains over forty different nutritional components such as
a wide range of enzymes; where these nutritional
components are in a good balance [56, 57]. In addition, raw
RJ has an unknown substance among its nutritional compo-
nents, which is called the “R substance” from the initial letter
of RJ.
Table 4. Composition of Royal Jelly (Takenaka, 54)
Minimum Maximum
Water 57% 70%
Proteins (N x 6.25) 17% of dry weight 45% of dry weight
Sugars 18% of dry weight 52% of dry weight
Lipids 3.5% of dry weight 19% of dry weight
Minerals 2% of dry weight 3% of dry weight
Recent Patents on Anti-Infective Drug Discovery, 2009, Vol. 4, No. 3 209
There is actual proof of its effects on bacterial infections
[58] (Table 5) and oxidative stress [59]. Products such as RJ
have been attracting attention as medical supplies and health
foods having the above-described effects, and they have
currently become standard health foods with consumptions
that increase each year. RJ consists of an emulsion of
proteins, carbohydrates, lipids, and other identified water-
soluble compounds. Proteins make up about 13 percent of
RJ, most of which belong to a family called major royal jelly
proteins. The antibacterial activity of RJ has been described
[7]; for instance, the RJ protein, royalisin, possesses
antibiotic properties against gram-positive, but not gram-
negative bacteria [8]. About 11 percent of RJ is sugars,
including fructose and glucose, similar to those found in
honey; lipids comprise about five percent of the substance
[60]. The potency of antibacterial properties of RJ might be
related to a particular fatty acid present in the ether-soluble
fraction of RJ called trans-10-hydroxy decanoic acid (10-
HDA) [8]. The characterization of novel antibacterial
peptides isolated from RJ, the jelleines, a series of short
peptides presenting broad-spectrum activity against gram-
positive bacteria, gram-negative bacteria, and yeasts, has
recently been described [7]. Due to the acidic taste of RJ, it
is usually mixed with honey to increase consumer
acceptance. The addition of honey also acts as a preservative
to permit the storage of RJ at room temperature for an
extended period of time [61]. A synergistic effect against
bacteria has been noticed when mixing RJ with starch [62].
RJ is also combined with other components to prevent the
skin from harmful bacteria [63].
PROPOLIS
Bees use propolis (bee glue) not only as a building
material but also to keep low concentration of bacteria and
fungi in the hive Fig. (3). Thus, the action against micro-
organisms is an essential characteristic of propolis and that’s
why it has been applied as a remedy by man since ancient
times [64]. Bees collect resins from numerous different plant
species, thus the chemical composition of propolis varies
greatly and depends directly on the local flora and phenology
of the host plants and indirectly on the locality and time of
collection. It is believed that part of the substances collected
undergoes chemical alteration by bee enzymes prior to
incorporation in propolis, making propolis a substance
considered to be of both plant and animal origin.
Specifically, bees mix the plant exudates (including resins,
waxes and oils collected from different parts of plants and
trees including branches, flowers, pollen and buds) with
beeswax which bees secrete through glands located near the
hypopharygeal region of the bees' bodies and
-glucosidase
they secrete during plant exudate collection and small
amounts of sugar, which results in whole Propolis [65].
While chemical composition varies, propolis is generally
composed of about 45% resin, 35% wax and 20% inert
material. More specifically, propolis constituents include
about 10% essential oils, 5% pollen, and 15% various
organic polyphenolic compounds including, flavonoids and
phenolic acids. Propolis is most often purified by extraction
with solvents, particularly ethanolic extracts of propolis. The
extraction process removes the wax and the inert material,
210 Recent Patents on Anti-Infective Drug Discovery, 2009, Vol. 4, No. 3
Fig. (3). Honeybees frequently use propolis to reduce the size of the
entrance for better defence (FAO, 60).
and preserves the resinous phenolic fraction. A multi-step
extraction with ethanol can result in dewaxed propolis
extracts rich in polyphenolic components [65]. There is
substantial evidence that propolis has antiseptic, antifungal,
antibacterial, antiviral, anti-inflammatory and antioxidant
properties [66]. The polyphenolic content of propolis is
considered to contribute more to the observed healing effects
than other propolis constituents. Propolis has a long history
of use in folk medicine [64]. Propolis antimicrobial property
has been widely investigated, and several authors have
demonstrated its antibacterial action [67-72]. Propolis also
shows antiviral [73, 74], antifungal [75, 76] and antiparasite
activities [77, 78]. Clinical findings are indicative that
propolis extracts may be effective in coccidiosis [79], tricho-
moniasis [80] and giardiasis [81]. Many therapeutic and
cosmetic formulations containing Propolis have been
described [82]. The oral preparations can be found in a form
of a mouth rinse, a dentifrice, a confectionary, a medicament,
or a film [83]. Propolis has been found to be effective against
oral pathogens [84] with high antibacterial activity against
Staphylococcus [85].
Table 5. The Inhibitory Activity of Ether-Soluble Fraction of
Royal Jelly (30 Mg/Ml) Against Bacteria Using Drop
Plate Method (Eshraghi and Seifollahi, [99])
Test Strain Inhibitory Zone
Diameter (mm)
Streptomyces strain (46) 23
Streptomyces strain (F8) 22
Streptomyces strain (66) 24
Streptomyces griseus (ATCC 11746) 29
Staphylococcus aureus (ATCC 14776) 40
Escherichia coli (ATCC 29532) 22
Boukraa and Sulaiman
BEE VENOM
Bee venom (BV) is a traditional medicine that has been
widely used with satisfactory results in the treatment of some
immune-related diseases, especially rheumatoid arthritis
(RA) [86]. BV contains various peptides such as melittin (the
major active ingredient of BV), apamin, adolapin, and mast
cell degranulating peptide [87] (Table 6). Several studies
have shown BV to exert both an anti-inflammatory effect, a
property shared with non-steroidal anti-inflammatory drugs
(NSAIDs) [88] and an antibacterial effect involving no side
effects in animal models [89]. Phospholipase A2 (PLA2) is
signifi-cantly involved in many cellular responses such as
phospholipid digestion and metabolism, host defence and
signal transduction [90-92]. Bee venom PLA2 (bvPLA2) is
easily obtained. BvPLA2 hydrolyses phospholipids of
erythrocyte and mast cell membranes and has been shown to
have antimicrobial activity on some Gram-negative bacteria
but there are no reports of the activity of purified bvPLA2
[93]. Melittin, a 26-residue peptide, present as the major
toxic component of the European bee venom, Apis mellifera
is characterized by its powerful hemolytic activity [94].
Although it possesses potent antimicrobial activity its
overriding cytotoxic effect makes it unsuitable for any
therapeutic use. Several reports have appeared describing
non-hemolytic analogs of melittin that retain their antimic-
robial activity. These analogs include retro, retro-enantio and
dia-stereo analogs and all of them are of the same length as
melittin [95]. Many studies and invention about allergen
immunotherapy have been described [96-99].
CURRENT & FUTURE DEVELOPMENTS
Natural products may have a large variance in therapeutic
components depending on their origin. Thus, the floral
source of honey plays an important role on its biological
properties. In consequence, it would not be surprising that
the provenance of honey could determine its antibacterial
properties. It is also possible that the mixing of bee products
affect their antibacterial activity since those with lower
antibacterial activities may mask the higher antibacterial
activity of others. Bee products that are to be used for
medicinal purposes have to meet certain criteria. As such,
they have to be free of residual herbicides, pesticides, heavy
metals, and radioactivity. They have also to be sterilized to
prevent secondary infections. The problem of antibiotic
residues should be highlighted. As long as beekeepers
continue to use antibacterial drugs to control bee diseases,
the risk of antibiotic residues in some bee products will
remain. Even authorities have allowed a Maximum Residues
Limit for each molecule, consumers want honey free of
residues because it is perceived as a pure, natural product.
Only bee products warranted antibiotic residues free should
be used for managing infections. Whichever bee product is
used for medicinal purposes, consideration needs to be given
to its quality. Further evidence and understanding of the
antibacterial properties of hive products are needed to
optimise their use in the clinical management of infections.
ACKNOWLEDGEMENTS
This review has been conducted under the scheme of
postdoctoral fellowship of Universiti Sains Malaysia.
Antibacterial Bee Products Recent Patents on Anti-Infective Drug Discovery, 2009, Vol. 4, No. 3 211

Table 6. Composition of Venom from Honeybee Worker (FAO, 60)

Class of Molecules Component % of Dry Venom

Enzymes Phospholipase A2 10-12

Hyaluronidase 1-3
Acid phosphomonoesterase
Lysophospholipase

-glucosidase

Other proteins and peptides Melittin 50

Pamine 1-3
Mast Cell Degranulating Peptide (MCD) 1-2
Secapin 0.5-2.0
Procamine 1-2
Adolapin
Protease inhibitor 0.1
Tertiapin 13-15
Small peptides (with less than 5 amino acids)

Physiologically active Histamine 0.5-2.0

amines Dopamine 0.2-1.0
Noradrenaline 0.1-0.5

Amino acids t -Aminobutyric acid 0.5

a-Amino acids 1

Sugars Glucose & fructose 2

Phospholipids 5

Volatile compounds 4-8

CONFLICT OF INTEREST
There is no conflict of interest.
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