ANTIMICROBIAL EFFECTS OF HONEY AND ITS SPECIFIC ACTIONS ON CELL WALLS, MEMBRANES AND ENZYMES OF SOME MICROBIAL PATHOGENS

ABSTRACT

Antimicrobial agents of plant origin have enormous therapeutic potentials. Honey, which is a product of plant, is a sugary substance produced by bee from the nectar of flower. It has been an age long antimicrobial therapy for wounds and burns. The aim of this study was to determine the antimicrobial effects of honey and its specific actions on cell walls, membranes and enzymes of the following organisms: Pseudomonas aeruginosa, Bacillus subtilis, Trichophyton verrucosum. Trichophyton equinum and Escherichia coli using agar well diffusion method. Pseudomonas aeruginosa, Bacillus subtilis, and Escherichia coli were tested at 100 %, 80 % and 60 % (v/v) honey concentration and Trichophyton verrucosum and Trichophyton equinum were tested at 100 % honey concentration only. Escherichia coli had the highest zone of inhibition of 29.0 mm, 27.0 mm and 19.0 mm at 100 %, 80 % and 60 % respectively followed by Bacillus subtilis, which had 15.0 mm, 1.0 mm and 8.0 mm zones of inhibition while Pseudomonas aeruginosa had 13.7 mm, 11.0 mm and 6.3 mm zones of inhibition as the least. At 100 %, T. verrucosum and T. equinum had zone of inhibition of 14.0 mm and 17.0 mm respectively. The minimum inhibitory concentrations recorded for Escherichia. coli, Bacillus subtilis and P. aeruginosa were 10 %, 80 % and 100 % (v/v) respectively. The minimum bactericidal concentrations for E. coli and B. subtilis were 20 % and 100 % (v/v) respectively. The effect of honey on bacterial isolates after incubation for one hour and two hours revealed that E. coli had 22.0 and 35.3 µg/ml protein leakage; Bacillus subtilis had 31.0 and 49.0 (µg/ml) while Pseudomonas. aeruginosa had 49.7 and 60.0 (µg/ml) respectively. The result of enzymatic inhibition showed that honey had activity against the cells treated compared to the control:  E. coli had 11.0 and 14.0 (mm); Bacillus. subtilis had 30.0 and 40.0 (mm) while Pseudomonas aeruginosa had 31.7 and 45.0 (mm) for the treated and untreated cells respectively. The result of this study showed that the honey had a broad spectrum antimicrobial activities and could be recommended for antibiotics alternative therapy.

CHAPTER ONE

1.0       INTRODUCTION

1.1       Background to the Study

Antimicrobial agents possess immense curative prospects and are very effective in controlling disease causing organisms (Mahato and Sharma, 2018). Prior to the discovery of microorganisms, it was generally believed that certain medicinal herbs had curative properties and, without a doubt, contained what we now refer to as antimicrobial principles (Mahato and Sharma, 2018). Common infectious diseases have been cured by the use of plants, and this form part of the customary management of different health challenges, some of these conventional medicines are still included. The rich sources of antimicrobial agents are medicinal plants (Jindal and Vashist, 2013).

Medicinal plants are rich sources of antimicrobial agents (Jindal and Vashist, 2013). According to World Health Organization (WHO), medicinal plants would be the best source to obtain a variety of drugs and 80 % of world population is dependent on traditional medicine and a major part of traditional therapies involves the use of plant extracts or their active constituents. Yet a scientific study to determine their antimicrobial active compounds is a comparatively new field. (Parmar and Rawat, 2012).

One of the earliest medicines used by traditionalist for both infectious and non-infectious diseases and was also recommended for management of burns and wounds was honey, a plant product according to the report of Olatunji et al. (2018).

Honey is a sugary and thick material formed by bees (Olatunji et al., 2018). It is made up of flower nectars (floral honey) and sweet plant deposits (non-floral honey), as well as an enzyme secreted by honeybees. These sugary substances are gathered by bees, who then improve them through their own materials before processing them in beehives (Manyi-Loh et al., 2011; Süerdem and Akyalçin, 2017).

Honey is a popular sweetener, non-toxic, non-irritant and a common household product (Usman et al., 2015). Ayurveda, an ancient Indian System of Health Care treats honey as food for health while recommending it as a medicine for some conditions using it externally as well as orally (Khandal et al., 2010). Ayurveda, which means science of long life, is believed to have originated over 6000 years ago and was designed to promote good health and long life rather than to fight disease and was practiced by physicians and surgeons (called Bheshaja or vaidya) but recently herbal medicine has attracted much attention as alternative medicines useful for treating or preventing life-style related disorders (Agyare et al., 2009).

Honey is a well-researched oldest medicine for a variety of pathogenic microbes, as well as an active antibacterial agent for burn injuries (Brudzynski, 2006). After topically applied to wounds, osmosis would be expected to draw water from the wound into the honey helping to dry the infected tissue and reduce bacteria growth (Al-Naama, 2009). Honey’s antibacterial properties have been demonstrated in various reports and clinical trials against a wide range of microorganisms, including multi-antibiotic drug resistant strains (Kumarasamy and Mahendran, 2015).

Laboratory studies and clinical trials have shown that honey is an effective broad- spectrum antimicrobial agent. Honey has been reported to have inhibitory effect on several bacteria including aerobes and anaerobes, Gram-positive and Gram-negative and is effective against methicillin resistant Staphylococcus aureus (MRSA), β-hemolytic Streptococci and vancomycin-resistant enterococci (VRE) (Allen et al., 2000; Kingsley, 2001).

The use of honey for wound infections treatment dated over 2000 years prior to bacterial discovery to be the cause of infections (Sushila et al., 2012). Olatunji et al. (2018) reported that the antimicrobial activity of honey was first recognized in 1892, and this was then accompanied by detailed studies to further substantiate this argument and to show factors leading to the activity of antimicrobials.

Natural medicinal products have been used for millennia in the treatment of multiple ailments (Manyi-Loh, et al., 2011). Although many have been superseded by conventional pharmaceutical approaches, there is currently, resurgence in interest in the use of honey and honey products by the populace. This choice of honey for therapeutic purpose is a branch of medicine called apitherapy (Ghosh and Playford, 2003).

Honey can be classified based on where the bee got resources for honey make up: Floral and non-floral honeys.  Floral honeys can either be unifloral or multifloral, depending whether the honey collected is from the nectar of the same flower or from nectar of flowers of various types (Manyi-Loh, et al., 2011). Non floral honey (honey dew) is made by bees that extract sugars from the living tissues of plants or fruits, and/or scavenge the excretions of insects (aphids) that tap the veins of higher plants (Subrahmanyam, 2007).

The antimicrobial effects of honey could be bacteriostatic or bactericidal depending on the concentration that is used. However, such activity has been attributed to certain factors like high osmolarity (low water activity), acidity (low pH), and hydrogen peroxide and non-peroxide components (Taormina et al., 2001; Al-Naama, 2009).

Among the possible therapeutic alternatives that are approved, non-toxic and with a wide range of antimicrobial spectrum of action, honey is considered. This may be a potential alternative or replacement for antimicrobial agents, but its use is constrained by certain factors.  Inadequate  knowledge  of  antimicrobial  properties  and  lack  of  adequate information have reduced the clinical applicability of honey (Malik et al., 2010; Mandal and Mandal, 2011).

In Nigeria, honey is accepted to be important in traditional treatment of respiratory ailments, skin infections, diarrhoea and other diseases as captured by Eleazu et al. (2013). There are numerous reports on the physico – chemical, antimicrobial, microbiological and medicinal properties of honey from many parts of the world, including North America, Europe, Asia, Australia, and South Africa (Gomes et al., 2010; Mandal and Mandal, 2011; Fahim et al., 2014).

Data on Nigerian honey is limited; however, some physical properties, antibacterial and chemical properties had been documented from honey in Nigeria (Adebiyi et al., 2004; Omafuvbe and Akanbi 2009; Anyanwu, 2012; Eleazu et al., 2013; Buba et al., 2013). Although, the antifugal study on Nigerian honey is minimal, nevertheless, separate study conducted by Akujobi and Njoku (2010), Anyanwu (2012), Eleazu et al. (2013) and Buba et al. (2013) indicated that Nigeria honey could kill fungi and also inhibit the growth of fungi

1.2       Statement of Research Problem

Attention had been drawn nowadays in serious search for antimicrobial compound all over the world owing to the failure of the already existing antibiotics (Ibrahim and Aliyu, 2015). With the irrational and massive use of antibiotics in underdeveloped and developing countries, resistance develops and spread beyond human imagination. As a result, the effectiveness of the antibiotics is reduced (Zakaria, 2015). Rural dwellers and disadvantaged people who are unable to have enough money to access good health care services relied on conventional medicines which they are familiar with for their treatment (Krishnan, 2018).

The World Health Organization (WHO) has assessed that up to 80 percent of individuals in the developing nations rely on local medicinal prescriptions because of their easy accessibility, wide affordability and cultural familiarity. Indeed, about 40 percent population of the world’s poor have no    good hospital, hence, they depend on local medicinal prescription (Krishnan, 2018).

Clinical acceptability of honey has been slowed down by curtailed knowledge of the antimicrobial activity and short accurate mechanisms for determine the type of action of honey and variations of honey (Malik et al., 2010). According to Elijah et al. (2015), the structural existence of honey may be linked to geographic origin, and the antibacterial function of floral sources may play a key role. In Nigeria, there is no comprehensive record that honey from all the States have antimicrobial activity and of course, Nigeria has different climatic zones

1.3       Justification for the Study

The antimicrobial information of traditional medicine is a panacea for battling antibiotic resistant microorganisms in the developing nation like Nigeria. Many traditional medicines had been hawked without antimicrobial authentication of which honey was part of. Moreover, the specific of action of any antimicrobial agents is very important as it gives specific information about its activity.

The antimicrobial activities of honey from different countries has been reported in recognition of the medicinal properties of honey (Olajuyigbe et al., 2017). In Nigeria, the therapeutic ability and  the antibacterial  activities  of various  honey samples  widely marketed for consumption need to be validated. Previous researched works on honey dwell on the antimicrobial activity

Therefore, for honey to be recommended as an alternative antimicrobial agent, like other conventional medicines, there is need for laboratory investigation on the specific action of honey on bacterial cell and its enzymes.

1.4       Aim and Objectives of the Study

The aim of this study was to determine the antimicrobial effects of honey and its specific actions on cell walls, membranes and enzymes of some microbial pathogens.

The specific objectives were to:

I          Assess the antimicrobial activity of honey.

II         Determine   the   minimum   inhibitory   concentration   (MIC)   and   minimum bactericidal concentration (MBC).

III        Determine the bioactive component of the honey.

IV       Determine the effects of honey on the cell walls, membranes and enzymes of the test organisms.

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