SUMMARY

Honey is a powerful antimicrobial agent with a wide range of effects. Various components
contribute to the antibacterial efficacy of honey: the sugar content; polyphenol compounds;
hydrogen peroxide; 1,2-dicarbonyl compounds; and bee defensin-1. All of these elements are
present at different concentrations depending on the source of nectar, bee type, and storage.
These components work synergistically, allowing honey to be potent against a variety of
microorganisms including multidrug resistant bacteria and modulate their resistance to
antimicrobial agents. Honey has long been recognized for its antimicrobial properties, and
numerous studies have investigated its efficacy against various bacterial species. This review is
intended to provide an overview of the Antibacterial activities of honey against some bacteria
species.

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INTRODUCTION

Honeybees make honey after feeding on flower nectar, blossoms, or by sucking on the flower’s

secretions. The collected substances are mixed together with other specific compounds from the

honeybees and are then deposited by the honeybees in the wax honeycomb and allowed to

mature over time. The composition of honey depends on the source of the plants that the bees

feed on (Eteraf-Oskouei and Najafi 2013, Schneider et al. 2013). However, honey of all origins

is composed mainly of the sugars glucose sucrose, and fructose, which constitute ~80% of its

weight, with water composing the remaining 20%. In addition, vitamins, flavonoids, amino

acids, enzymes, minerals, and phenolic acids are also present in honey. Honey has anti-

inflammatory (Tonks et al. 2016), healing (Bergman et al. 2019), antioxidant, and antineoplastic

effects (Irish et al. 2016).

The effectiveness and potency of honey against microorganisms depends on the type of the

honey produced, which is contingent on its botanical origin, the health of the bee, its origin, and

processing method. The antioxidant activity of honey is very much dependent on the botanical

origin where the bee was reared (Chauhan et al., 2010). The purpose of this review is to provide

an overview of the antibacterial activity of honey, the underlying factors that influence its

antibacterial efficacy, and its applications.

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HONEY

Honey is a natural substance produced by honeybees (Apis mellifera) from the nectar of flowers

which is a sweet, flavourful, viscose liquid (Dessie, 2021). It has been used as a food and

medical product since the earliest time (Herringtone, 2016). Honey is produced in almost every

country of the world and is a good energy food (James et al., 2019). Because of its high

nutritious value and unique flavour, honey has become increasingly accepted by consumers,

often being used as a substitute for other sweeteners (Mann et al., 2017). Honey is not to be

considered a whole food according to human standards but rather as a potential dietary

supplement (Olugbemi et al., 2013).

Figure 1: Honey Comb

Source: (Olugbemi et al., 2013).

SOURCES OF HONEY

Carbohydrate

Honey is mainly made up of carbohydrates (82.3%) which constitute about 95% of its dry weight

and is a highly complex mixture of sugars. The major sugars in honey are Monosaccharide

consisting of fructose (38%) and glucose (31%) (Alvarez-Suarez et al., 2019), which are

responsible for most of the physical and nutritional characteristics of honey. (Lawal et al., 2019).

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Proteins, Enzymes and Amino Acids

Honey naturally contains small amount of enzymes that are introduced into honey by the bees

during various phases of the honey manufacturing process. The three main honey enzymes are

diastase (amylase), decomposing starch or glycogen into smaller sugar units, 8 invertase

(sucrase, α-glucosidase), decomposing sucrose into fructose and glucose, and glucose oxidase,

producing hydrogen peroxide and gluconic acid from glucose. The amino acids identified in

honey from different botanical and geographical origin are: glutamic acid, aspartic acid,

glutamine, histidine, glycine, threonine, β-alanine, arginine, α-alanine, γ-aminobutyric acid,

proline, tyrosine, valine, methionine, cysteine, isoleucine, leucine, tryptophan, phenylalanine,

ornithine and lysine (Saba et al., 2013).

Vitamins, Minerals Compounds Present in Honey

Honey contains varying amounts of vitamins, minerals and trace elements. The vitamins content

in honey is low. However, vitamins such as phyllochinone (K), thiamine (B1), riboflavin (B2),

niacin (B3), pantothenic acid (B5) and pyridoxine (B6) as well as vitamins A, E and C have been

reported to be present in honey. The mineral content of honey depends mainly on the botanical

and geographical origin of honey. According to (Saba et al., 2013), the minerals commonly

found in honey are potassium, aluminium, barium, nickel, chromium, cobalt, calcium, iron, zinc,

magnesium, copper, sodium and manganese.

TYPES OF HONEY

Honey can be classified based on its floral source, colour, flavour, region where it was produced

(regional honeys) or according the packaging and processing used. (Manyi-Loh et al., 2011).

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Floral Sources

Honey can be classified based on the source of nectar for which it was produced (Manyi-Loh et

al., 2011). These differences may be as a result of the quantity and quality of materials such as

pollen, nectar etc. used in honey production (Manyi-Loh et al., 2011).

Types of Honey Based on Floral Source

Monofloral Honey: This honey is made primarily from the nectar of one type of flower.

Different monofloral honey has a distractive flavour and colour because of differences between

their principal nectar sources (Manyi-Loh et al., 2011).

Polyfloral Honey: This is honey got from the nectar of many types of flowers.

Blended Honey: This is a mixture of two more honey differing in floral sources, colour, flavour,

density or geographical origin (Manyi-Loh et al., 2011).

PROPERTIES OF HONEY

The properties of honey vary depending on water content, the type of flora used to produce it,

temperature and the proportion of the specific sugars it contains. Fresh honey is a supersaturated

liquid, containing more sugar than the water can typically dissolve at ambient temperatures. At

room temperature, honey is a super cooled liquid, in which the glucose will precipitate into solid

granules. This forms a semisolid solution of precipitated glucose crystals in a solution of fructose

and other ingredients (Alvarez-Suarez et al., 2019).

Phase Transitions

The melting point of crystallized honey is between 40 and 50 °C (104 and 122 °F), depending on

its composition. Below this temperature, honey can be either in a metastable state, meaning that

it will not crystallize until a seed crystal is added, or, more often, it is in a "labile" state, being

5
saturated with enough sugars to crystallize spontaneously. Therefore, larger but fewer crystals

tend to form at higher temperatures while smaller but more-numerous crystals usually form at

lower temperatures. Below 5 °C, the honey will not crystallize and thus, the original texture and

flavour can be preserved indefinitely (Dessie, 2021).

Viscosity

Since honey normally exists below its melting point, it is a super cooled liquid. The viscosity of

honey is affected greatly by both temperature and water content. The higher the water

percentage, the easier honey flows. Aside water content, the composition of honey also has little

effect on viscosity, with the exception of a few types. At 25 °C (77 °F), honey with 14% water

content generally has a viscosity around 400 poise, while a honey containing 20% water has a

viscosity around 20 poise. At very low temperatures, honey will not freeze solid (Dessie, 2021).

Electrical and Optical Properties

Because honey contains electrolytes, in the form of acids and minerals, it exhibits varying

degrees of electrical conductivity. Measurements of the electrical conductivity are used to

determine the quality of honey in terms of ash content (Lawal et al., 2019).

The effect honey has on light is useful for determining the type and quality. Variations in the

water content alters the refractive index of honey. Water content can easily be measured with a

refractometer. Typically, the refractive index for honey will range from 1.504 at 13% water

content to 1.474 at 25%. Honey also has an effect on polarized light, in that it will rotate the

polarization plane. The fructose will give a negative rotation, while the glucose will give a

positive one. The overall rotation can be used to measure the ratio of the mixture (Saba et al.,

2013).

Thermal Characteristics

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Like all sugar compounds, honey will caramelize if heated sufficiently, becoming darker in

colour, and eventually burn. However, honey contains fructose, which caramelizes at lower

temperatures than the glucose. The temperature at which caramelization begins varies, depending

on the composition, but is typically between 70 and 110 °C (158 and 230 °F). Honey also

contains acids, which act as catalysts, decreasing the caramelization temperature even more. Of

these acids, the amino acids, which occur in very small amounts, play an important role in the

darkening of honey. The amino acids form darkened compounds called melanoidins, during a

Maillard reaction (Nzeakor and Hamdi, 2020)

PARAMETERS RELATED TO THE ANTIMICROBIAL ACTIVITY OF HONEY

Low water activity

Water is a major constituent of living creatures and is found in various foods in the form of free

or bound molecules. In honey, water activity measures unbound water molecules and ranges

from 0.562 to 0.62, a concentration low enough to allow the growth of bacteria or other

microorganisms (Molan 2012).

High sugar content

Osmosis is induced as a result of the high sugar concentration in honey. Pure, undiluted honey

therefore inhibits the growth of bacteria due to its sugar content, which exerts osmotic pressure

on bacterial cells, causing water to flow out of the bacterial cells via osmosis. As a result, the

cells shrink due to dehydration and are unable to survive in the hypertonic sugar solution. Such

antibacterial potential will be lower when honey is mixed with bodily fluids at infection sites

(Molan 2012).

Acidity

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The optimal growth of most microorganisms occurs at neutral pH, ranging from 6.5 to 7.5. The

acidity of honey, between pH 3.2 and pH 4.5, is a very marked characteristic of its antibacterial

efficacy. This acidity is caused by the presence of certain important organic acids, especially

gluconic acid, found at a concentration of at ∼0.5% (w/v). Glycogenic acid is generated from

glucose oxidation by an endogenous glucose oxidase enzyme and is an extremely potent

antibacterial agent. In undiluted pure honey, the low pH may contribute to its antibacterial

potency, however, pH alone is not sufficient to inhibit the growth of many types of bacteria when

diluted in food or in other bodily liquids (Molan 2012).

Hydrogen peroxide

Hydrogen peroxide (H2O2) is a disinfectant and a strong oxidizing agent (Ali, 2014). It provides

honey with its antibacterial efficacy and is produced enzymatically (Figure 3). The enzyme

glucose oxidase is naturally present in an inactive state in honey due to the low pH conditions.

When honey is diluted, glucose oxidase is activated and acts on endogenous glucose to produce

H2O2. In fact, the maximum level of hydrogen peroxide can be achieved by diluting honey by

30%–50%, typically in the range of 5 to 100 μg H 2O2/g honey (equivalent to ∼0.146–2.93 mM)

(Brudzynski 2016)

Proteins

To date, there have been very few published papers on the protein content of honey (Chua et al.

2013). Honey has relatively tiny amounts of proteins, ranging from ~0.1%– 0.5%, with

molecular weights ranging from 20 to 80 kDa (Tewari and Irudayaraj 2014). Those proteins

include many enzymes involved in sugar metabolism, such as alpha and beta glucosidase,

glucose oxidase, and amylase (Kwakman et al. 2011).

8
Figure 2. Schematic diagram showing the parameters that contribute to the antimicrobial
potential of honey
Source: (Tseng et al. 2011).

ANTIBACTERIAL EFFECT OF HONEY

The antibacterial efficacy of honey was initially recognized in 1892; however, in modern

medicine, it is used only to a limited extent due to the absence of scientific support (Mohapatra

et al. 2011).

Honey exhibits antibacterial activity against numerous bacteria in different environments. The

natural components of honey have various activities against different microorganisms (Figure 3).

The antibacterial activity of honey is likely to depend on the pasture on which the bees were

raised, climatic conditions, as well as the natural composition of the flower nectar (Abd-El Aal

et al. 2017).

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Figure 3. Schematic diagram showing the range of antimicrobial activities of honey.
Source: (Mohapatra et al. 2011).

Honey has excellent antibacterial efficacy against MRSA and a variety of Pseudomonas, which

are often associated with wound and burn infections (Hazrati et al. 2010). Manuka (L.

scoparium) honey (Visavadia et al., 2018) showed efficacy against many pathogenic

microorganisms, including: Enterobacter erogen, S. aureus, Salmonella zyphimurium, and last,

but not least, Escherichia coli (E. coli) (Lusby et al. 2015). Many studies revealed that honey

was effective against MRSA, hemolytic streptococci, and vancomycin-resistant enterococci

(Figure 4) (Lusby et al. 2015). However, newly recognized local honey types may be as good as

or even better than Manuka honey based on their impressive antimicrobial activity.

Unlike Manuka honey, the activity of Almo honey is mainly due to its production of H2O2. A

25% solution (v/v) of Almo honey did not exhibit any antibacterial activity in the presence of the

catalase enzyme, which scavenges H2O2 activity. This is in contrast to Manuka honey, which

maintained its antibacterial potential under the same conditions (Sherlock et al. 2010).

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Sterilization by gamma irradiation does not interfere with Almo honey activity (Simon 2009).

The pH of honey is low enough to cause inhibition of certain bacterial pathogens, like S.

pyogenes (4.5), E. coli (4.3), P. aeruginosa (4.4), and Salmonella spp. (4.0) and, as such

therefore, acidic honey is an important antibacterial factor (Haniyeh et al. 2010).

Manuka honey can modulate bacterial size and shape, which affects the septal ring involved in

cell division (Lu et al. 2013). Henriques et al. (2010) used transmission electron microscopy to

observe the effects of Manuka honey on S. aureus cultures and showed that more septal cells

were found in the samples treated with Manuka honey in comparison to those treated with

artificial honey made from sugars and water that mimic in its composition the natural honey. A

study using phase contrast imaging found that Bacillus subtilis and S. aureus bacteria treated

with a sub-lethal dose of Manuka honey (4%, w/v) resulted in the appearance of smaller cells

containing condensed chromosomes (Tonks et al. 2013). Interestingly, both (Henriques et al.

2010) and (Lu et al. 2013) showed that the stress caused by honey resulted in up-regulation of

cell growth and affected bacterial cell division.

Figure 4. Schematic representation of the antimicrobial effects of honey.

Source: (Henriques et al. 2010).

Studies by (Al-Nahari et al. 2015, Almasaudi et al. 2017) evaluated the efficacy of different

types of Manuka honey (UMF 10, 16, and 20) in addition to two types of Saudi honey (Sidr and

11
Nigella sativa) against Gram-positive methicillin resistant and sensitive S. aureus as well as

Gramnegative imipenem-resistant and sensitive Pseudomonas aeruginosa. The results showed

that all bacterial strains were completely inhibited at honey concentrations between 10% and

50%, depending on the strain and the efficacy of the type of honey used. Manuka UMF-20 was

superior by far to all other types of honey tested and produced a bactericidal effect, whereas the

Saudi honeys were bacteriostatic (Al-Nahari et al. 2015, Almasaudi et al. 2017).

ANTIBIOTIC SYNERGISM WITH HONEY

Multidrug-resistant (MDR) bacteria are a global problem, and infections by resistant bacteria are

on the rise in various animal species, including humans (Szmolka and Nagy, 2013). For this

reason, there is a major effort to find alternative therapies with better efficacy and to reduce

antibiotic use and combat antibiotic resistance. Alternative therapies may be particularly useful

in mild infections. The antibacterial properties of honey have long been known, and honey

exhibits broad activity against MDR bacteria. For this reason, many studies have been carried out

to evaluate the efficacy of the application of honey with antimicrobial agents, with some positive

outcomes (Szmolka and Nagy, 2013).

The application of tetracycline with Manuka honey yielded better antimicrobial potential against

S. aureus and P. aeruginosa than was observed with either treatment alone. This finding suggests

that such a combination is a possible treatment strategy for wound healing (Jenkins and Cooper

2012). In a different study, the combination of rifampicin with sub inhibitory concentrations of

Medihoney reversed rifampicin resistance in clinical isolates of S. aureus, including MRSA

(Müller et al., 2013). Other data also support that the application of honey together with

antibiotics can modulate antibiotic resistance. For instance, (Jenkins and Cooper 2012) reported

12
that MRSA became susceptible to oxacillin upon the application of sub inhibitory concentrations

of honey.

The emergence of bacterial resistance to honey is very low because of variations in its

composition due to differences in: 1) nectar source; 2) climatic conditions; 3) duration of storage;

and 4) preservation conditions (Sherlock et al. 2010).

Moreover, synergistic effects were noted against biofilms using combinations of honey and

antibiotics. This was shown for the use of Manuka honey with vancomycin against S. aureus and

for the combination of Manuka honey with gentamicin against P. aeruginosa (Campeau and

Patel 2014). Additionally, a reported synergism between Portuguese honey and phage therapy

showed that 25% (w/v) honey caused synergism with phage and was also efficient in the

eradication of E. coli biofilms at a 50% (w/v) honey alone (Oliveira et al. 2017).

ANTIBACTERIAL ACTIVITIES OF HONEY AGAINST SOME BACTERIA SPECIES

Honey has been recognized for its antimicrobial properties for centuries, and numerous studies

have investigated its efficacy against various bacterial species. Here are some common bacteria

species against which honey has shown antibacterial activity:

Staphylococcus aureus (S. aureus):

S. aureus is a common bacterium responsible for various infections, including skin infections,

wound infections, and food poisoning. Honey, particularly Manuka honey, has been found to

exhibit potent antibacterial activity against S. aureus strains, including methicillin-resistant S.

aureus (MRSA) (Müller et al., 2013).

Escherichia coli (E. coli):

E. coli is a bacterium commonly found in the intestines of humans and animals. Certain strains of

E. coli can cause foodborne illnesses and urinary tract infections. Several studies have reported

13
the antibacterial activity of honey against E. coli, inhibiting its growth and reducing its viability

(Sherlock et al. 2010).

Pseudomonas aeruginosa:

P. aeruginosa is a pathogenic bacterium associated with various infections, particularly in

individuals with compromised immune systems or cystic fibrosis. Honey has been shown to

inhibit the growth of P. aeruginosa and reduce its virulence factors, making it a potential

therapeutic agent against Pseudomonas infections (Campeau and Patel, 2014).

Salmonella spp.:

Salmonella species are a common cause of foodborne illness, leading to symptoms such as

diarrhea, fever, and abdominal cramps. Studies have demonstrated the antibacterial effects of

honey against Salmonella spp., suggesting its potential as a natural antimicrobial agent in food

safety applications (Szmolka and Nagy, 2013).

Helicobacter pylori (H. pylori):

H. pylori is a bacterium associated with gastrointestinal disorders, including peptic ulcers and

gastritis. Some research suggests that certain types of honey may possess antibacterial activity

against H. pylori, potentially contributing to the management of H. pylori-related gastrointestinal

conditions (Carnwath et al., 2014).

Streptococcus spp.:

Streptococcus species are Gram-positive bacteria commonly found in the human respiratory tract

and oral cavity. Some strains can cause infections such as strep throat and dental caries. Honey

has been shown to exhibit inhibitory effects against various Streptococcus species, suggesting its

potential as a natural remedy for oral and respiratory infections (Campeau and Patel, 2014).

14
CONCLUSION

In conclusion, honey has demonstrated significant antibacterial activity against a range of

bacterial species, including clinically relevant pathogens such as Staphylococcus aureus,

Escherichia coli, Pseudomonas aeruginosa, Salmonella spp., Helicobacter pylori, and

Streptococcus spp. These findings suggest that honey holds promise as a natural antimicrobial

agent with potential applications in various fields, including medicine, food safety, and oral

hygiene. The antibacterial effects of honey are attributed to its complex composition, which

includes sugars, organic acids, enzymes, and phytochemicals with antimicrobial properties.

Additionally, honey's low pH, high osmolarity, and production of hydrogen peroxide contribute

to its antimicrobial activity.

Overall, the antibacterial activities of honey represent an exciting area of research with potential

implications for combating bacterial infections and promoting public health. Further exploration

of honey's antimicrobial properties may lead to the development of novel therapeutic strategies

and the utilization of honey as a complementary approach to conventional antibacterial agents.

RECOMMENDATIONS

i. Consider using honey types known for their potent antibacterial properties, such as

Manuka honey.

ii. Explore the use of honey-based dressings or topical applications for wound care,

particularly for the treatment of infected wounds, burns, and ulcers.

iii. Explore the use of honey as a natural preservative or antimicrobial agent in food products

to improve food safety and shelf life.

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iv. Ensure quality control measures are in place to maintain the consistency and efficacy of

honey-based products.

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