ABSTRACT
Honey has been used for various purposes including treatment of
some diseases locally especially in wounds
and upper respiratory tract infections (URTIs). There is paucity of information on the
scientific basis for the use of honey in the treatment of upper respiratory
tract infections caused by bacteria. This study was carried out to determine
antibacterial activities of honey on upper respiratory organisms. The
antimicrobial activities of the honeys were assessed against Streptococcus
pyogen, Pseudomonas aeruginosa Klebsiella
pneumoniae and Streptococcus
pneumoniae using agar well
diffusion technique. Broth dilution method was used to determine the minimum
inhibitory concentration (MIC) of the honey against the bacteria. Screening
concentrations of neat honey and 80% honey inhibited Streptococcus pyogen and
Klebsiella pneumoniae. Pseudomonas aeruginosa was resistant to
all concentrations of honeys except that inhibited the organism at 100%
concentration. Orthodox antibiotics showed significantly higher mean zone of
inhibition in relation to honey (P < 0.001). The sensitive organisms were
inhibited in dose related manner. The mean MIC of the different honey was
generally at 31.25% against Staphylococcus aureus while concentration of
50% showed bactericidal effect on Streptococcus pyogen. Klebsiella
pneumoniae was inhibited at mean concentration of 31.5% and bactericidal
effect was observed against the organism at 40% mean concentration.
TABLE OF CONTENTS
CHAPTER
ONE
1.0 Introduction
1.1 Antibacterial
Activity of Honey
1.2
Aims and Objectives
1.3
Objectives
CHAPTER
TWO
2.0
LITERATURE
REVIEW
2.1
Causative Agents of Upper
Respiratory Infection
CHAPTER
THREE
3.0 MATERIALS AND METHODS
3.1 Sample
Collection
3.2
Test Microorganisms
3.3 Media Used
3.4
Sterilization
3.5 Microbiological
Analysis
3.6 Identification
Of Bacterial Isolates
3.6.1
Gram
Staining
3.7 Biochemical
Cultural Characteristics
3.7.1 Catalase Test
3.7.2 Coagulase
Test
3.7.3 Citrate
Test
3.7.4 Motility,
Indole, Urease Test (Miu).
3.7.5 Triple
Sugar Iron Test
3.7.6 Oxidase
Test
3.8
Antibacterial Activity Assay
3.9 Determination
Of Minimum Inhibitory Concentration (Mic)
CHAPTER
FOUR
4.0 RESULTS
CHAPTER
FIVE
5.0
DISCUSSION, CONCLUSION AND RECOMMENDATION
5.1 Discussion
5.2 Conclusion
5.3 Recommendation
References
LIST OF TABLES
Table 1: Shows
the Antibiotic Sensitivity test of
Bacteria isolated expressed by its Diameter of
inhibition zone.
Table 2: Minimum inhibitory
concentration of Honey against gram positive upper respiratory
organisms.
Table 3: Minimum
inhibitory concentration of Honey against gram negative upper respiratory
organisms.
CHAPTER ONE
1.0 INTRODUCTION
Honey
is one of the oldest traditional medicines considered as traditional remedy for
microbial infections. It is also recognized as an efficacious topical
antimicrobial agent in the treatment of burns and wounds (Brudzynski,2006).
This leads to the search for different types of honey with antibacterial
activity (Mullai and Menon,2007). The healing effect of honey could be due to
various physical and chemical properties (Snow and Manley-Harris, 2004). The
floral source of honey plays an important role on its biological properties
(Molan,2002).
Honey is being used in a few hospitals,
especially in the clinical treatment of ulcers, bedsores, burns, injuries and
surgical wounds. The antibacterial properties of honey may be particularly
useful against bacteria which have developed resistance to many antibiotics,
e.g. Staphylococcus aureus, which is a major cause of wound sepsis in
hospitals (Armstrong and Otis, 1995). Honey is thus an ideal topical wound
dressing agent in surgical infections, burns and wound infections (Betts and
Molan, 2002).The use of honey as a medicine has continued into the present-day
medicine. It has been shown that natural unheated honey has some broad-spectrum
antibacterial activity when tested against pathogenic bacteria, oral bacteria
as well as food spoilage bacteria (Bassom et al., 1994, Mundo et al.,
2004 and Lusby and Coombes 2005).
The antibacterial potency of honey has been
attributed to its strong osmotic effect, naturally low PH (Kwakman and Zaat,
2012), the ability to produced hydrogen peroxide which plays a key role in the
antimicrobial activity of honey (Kacaniova et al., 2011 and
Wahdam,1998)and phytochemical factors. Numerous reports and clinical studies
have demonstrated the antimicrobial activity of honey against a broad range of
microorganisms, including multi-antibiotic resistant strains. Others studies
demonstrated the antibacterial activity of honey against: Escherichia coli,
Campylobacter jejuni, Salmonella entercolitis, Shigella dysenteriae (Adebolu,
2005 and Voidaou et al., 2011), Mycobacterium (Asadi-Pooya et
al.,2003), Methicillin-resistant Staphylococcus aureus and
Vancomymin-resistant Enterococci (Cooper et al.,1999 and 2002 and
Al-waili et al.,2005),Common gastrointestinal pathogenic bacteria(Lin et
al.,2011),and the development of streptococcus pyogenes biofilms
(Maddocks et al., 2012). The antifungal activity of the honey,
especially anti-Candida activity (Irish et al., 2006, Koc et al.,
2008 and Ahmad et al., 2012).
Honey is composed of approximately 82.4%
total carbohydrates (38.5% fructose, 31.0% glucose and 12.9% from carbohydrates
consisting of maltose, sucrose and other sugars) (Khan et al., 2007; Vallianou
et al., 2014). The natural ingredients of honey show different activities
against various microorganisms.
It’s activity is likely to be dependent on
the grazing grounds and the weather conditions where the bees were raised, and
on the natural structure of the blossom nectar (Abd-El Aal et al., 2007). Honey has an increasing effect on the levels of
anti-oxidants, iron and rare elements in blood (Theunissen et al., 2001). Abd-El Aal et
al. (2007) showed that honey had a more pronounced inhibitory effect
(85.7%) on Gram negative bacteria (Pseudomonas
aeruginosa, Enterobacter spp., Klebsiella)
in comparison to commonly used antimicrobial agents. A 100% inhibition was
observed in the case of Gram positive methicillin-resistant Staphylococcus aureus in comparison to
the use of antibiotics alone. A synergistic effect was achieved upon the
application of honey together with the antimicrobial agents in both Gram
negative and positive bacteria. Al Somal et
al. (1994) reported the inhibitory effect of Manuka Honey on Helicobacter pylori growth. In addition,
it was documented that honey could completely heal severe injuries (Visavadia et al., 2008).
The use of honey as a drug for the treatment
of disease dates back to 2100-2000 BC. For instance, pale honey was described
by Aristotle (384-322 BC) as being ‘‘good for sore eyes and wounds” (Mandal and
Mandal, 2011; Vallianou et al.,
2014). The antimicrobial properties of honey have been well documented, and
honey has been used from ancient times as a method of accelerating wound
healing. Its potential to assist wound healing has been demonstrated repeatedly
(Molan, 1999; Vallianou et al.,
2014). A possible reason behind its activity relies on its ability to generate
hydrogen peroxide by the bee-derived enzyme glucose oxidase (Saleh et al.,
2011; Jing et al., 2014). Another
possibility is the composition of honey, which has more than 181 constituents
(Bogdanov and Martin, 2002; Gheldof et
al., 2002; Mandal and Mandal, 2011; Vallianou et al., 2014). Staphylococci
bacteria are Gram-positive cocci (Ryan and Ray, 2004). The genus Staphylococcus is composed of 33 species
(Bergey and Holt, 1994). Most staphylococci
constitute the normal flora of the skin and mucus membranes (Madigan, 2005).
Some are aerobic while others are anaerobic and can grow at high salt
concentrations, reaching up to 10% (Murray et
al., 2005). The most pathogenic species is S. aureus (Murray et al.,
2005). Some coagulase-negative staphylococci (CNS) strains, causative agents of
infection in immune compromised individuals, developed resistance to
antibiotics.
These bacteria colonize devices that are
implanted in the human body, such as nails, slides and industrial joints used
in bones, heart valves and catheters of various types, as well as in peritoneal
dialysis. It has been observed lately that there was an increase in the
prevalence and incidence of methicillin resistant CNS and S. aureus, making it
more challenging to treat such infections (Kloos and Bannerman, 1994).
Coagulase-negative staphylococci are considered one of the most prevalent
microorganisms that are involved in hospital acquired infections (Tunney et al., 1996). Honey has been used to
inhibit these bacteria as well as to prevent and treat skin and other
infections (French et al., 2005).
Honey
has been used as a medicine since ancient times in many cultures and is still
used in „folk medicine‟. The use of honey as a therapeutic substance has been
rediscovered by the medical profession in more recent times, and it is gaining
acceptance as an antibacterial agent for the treatment of ulcers and bed sores,
and other infections resulting from burns and wounds. In many of the cases in
the cited reports, honey was used on infections not responding to standard
effective in rapidly clearing up infection and promoting healing. Honey has
also been found to be effective in treating bacterial gastroentertis in infants
(Molan and Peter,2006).
1.1
ANTIBACTERIAL ACTIVITY OF HONEY
The use of honey as a traditional remedy for
microbial infections dates back to ancient times (Molan, 1992). Research has
been conducted on manuka (L. scoparium) honey which has been demonstrated to be
effective against several human pathogens, including Escherichia coli (E. coli), Enterobacter aerogenes, Salmonella typhimurium, S. aureus (Lusby et al.,2005). Laboratory studies have
revealed that the honey is effective against methicillin-resistant S. aureus (MRSA), B-haemolytic streptococci
and vancomycin resistant Enterococci (VRE) (Allen et al.,2000).
However, the newly identified honeys may have
advantages over or similarities with manuka honey due to enhanced antimicrobial
activity, local production (thus availability), and greater selectivity against
medically important organisms (Lusby et al.,2005). The coagulase negative staphylococci are very
similar to S. aureus in their
susceptibility to honey of similar antibacterial potency and more susceptible
than Pseudomonas aeruginosa (P. aeruginosa) and Enterococcus species(Cooper et al.,2000).
The
antibacterial activity of honey was first recognized in 1892, by van Ketel
(Dustmann, 1979). Honey is produced from many sources and its antimicrobial
activity varies greatly with origin and processing (Molan, 1992). Honey has
been used as a medicine in many cultures for a long time (Quinn et al., 1994). It has been rediscovered
by the medical profession and it is gaining acceptance as an antibacterial
treatment of topical infections resulting from burns and wounds (Abuharfeil et al., 1999). Numerous studies
demonstrate that honey possesses antimicrobial activity (Dustmann, 1979; Molan,
1992). More recently, honey has been reported to have an inhibitory effect to
around 60 species of bacteria including aerobes and anaerobes, gram-positives
and gram-negatives (Molan, 1992), it destroys and/or inhibits the growth of
some pathogenic vegetative micro-organisms (Chick and Shin, 2001). An
antifungal action has also been ob-served for some yeasts and species of Aspergillus and Penicillium (Quinn et al.,
1994), as well as all the common dermatophytes (Brady et al., 1997).
Honey
possesses inherent antimicrobial properties, some of which are due to high
osmotic pressure or low water activity, in which the low water activity of
honey is inhibitory to the growth of the majority of bacteria and to many
yeasts and moulds. When applied topically to wounds, osmosis would be expected
to draw water from the wound into the honey, helping to dry the infected tissue
and reduce bacterial growth. Even when diluted with water absorbed from wounds,
honeys would be likely to retain a water activity sufficiently low to inhibit
growth of most bacteria. Honey is mildly acidic, with a pH between 3.2 and 4.5,
gluconic acid is formed in honey when bees secrete the enzyme glucose oxidase,
which catalyses the oxidation of glucose to gluconic acid, the low pH alone is
inhibitory to many pathogenic bacteria and, in topical applications at least,
could be sufficient to exert an inhibitory effect (Molan, 1995). Hydrogen
peroxide, the end product of the glucose oxidase system and tetracycline
derivatives has the antibacterial properties against pathogens (Snowdon and
Cliver, 1996). Low concentrations of this known antiseptic are effective
against infectious bacteria and can play a role in the wound healing mechanism
(Molan, 2001) and in stimulation and proliferation of peripheral blood
lymphocytic and phagocytic activity (Tonks et
al., 2001). Other factors, such as low protein content, high carbon to
nitrogen ratio, low redox potential due to the high content of reducing sugars,
viscosity/anaerobic environment and other chemical agent or phytochemicals are
also likely to play some role in defining antibacterial activity of honey
(Honey, 2002). Furthermore, honey has
been employed to shorten the duration of diarrhea in patients with bactericidal
gastro-enteritis due to bacterial infection (Haffejee and Moosa, 1985).
However, honey has other important beneficial characteristics that are less
influenced by storage conditions (Cooper et
al., 2002).
1.2 AIMS AND OBJECTIVES
This
study aims to evaluate the antibacterial activity of honey against pathogenic
organisms causing upper respiratory tract infections.
1.3 OBJECTIVES
1. To
determine the antimicrobial effect of honey on upper respiratory organisms.
2.
To determine the minimum inhibitory concentration of honey .
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