Medical jobs have been originating in the direction of infective diseases due to the widespread development of extremely immune micro-organism. This microbic opposition is attributed to the huge usage of antibiotics.
The outgrowth of drug opposition prompted the involvement of the research workers in the usage of readily available alternate medicative therapy to command the infection in order to forestall the over usage of antibiotics.
Recently, there are reappraisals and researches conducted on the usage of honey. Its usage in advancing lesion healing has long been known since antediluvian times. In fact, it is mentioned in one of the poetries of Holy Qur’an imputing curative values to it.
Nowadays, there are several probes conducted sing its antibacterial activity. However, at present, there has been no local survey yet in measuring if there is an antibacterial consequence of the locally available organic flowered honey in Toril, Davao City on the common pathogens in ague and chronic bacterial sinusitis. An in-vitro probe of the authority of honey is thought to be a necessary measure before its application in the intervention sinonasal infections.
Review of Related Literature: “ What is already known about the subject? ”
Honey
Honey is prepared by bees from works nectars, from works secernments and from eliminations of works sucking insects ( “ honeydew ” ) . The Food Standards Code defines it as “ the nectar and cloying exudates of workss gathered, modified and stored by the honey bee ” . The British formulary ( 1993 ) defines purified honey as being “ obtained by purification of the honey from the comb of the bee, Apis mellifera L, and other species of Apis ( Office of Complementary Medicine, 1998 ) .
Honey is a rich beginning of saccharides chiefly fructose and glucose. The chemical composing of honey varies depending on the works beginning, season and production methods. Therefore the colour, concentration and compounds vary depending on the flowered beginnings. Other compounds which can be found in honey include proteins and acids such as gluconic acid ( C6H11O7, besides known as 2,3,4,5,6-pentahydroxyhexanoic Acid ) , Minerals and Anti-Oxidants such as H peroxide ( H202 ) and Vitamins ( B6 and B12 ) ( BD Yates et Al, 1996 ) .
Honey has a low pH and a low wet content, which is normally on norm about 17 per centum. The gluconic acid in honey is produced when bees secrete glucose oxidase, while treating the nectar, this give honey a low pH. There are many assortments of honey from around the universe which come in three chief types which are liquid, whipped and comb honey ( Office of Complementary Medicine, 1998 ) .
Honey has long been known for its medicinal belongingss since antediluvian times ( Dustmann, 1979 ) . Its healing intents are mentioned in the Bible, the Koran, and the Torah. Its medicative application was known but nil much on its antibacterial activity.
The antibacterial action of honey was recognized for the first clip in 1892 by Van Ketel. It has been assumed that this is due wholly to the osmotic consequence of its high sugar content ( Molan, 2001 ) . Recently, its antibacterial belongingss have been extensively explored. The mechanism of antibacterial activity of honey is attributed to its following natural belongingss: low pH, H peroxide, and low H2O activity. A low pH is repressive to most bacteriums since largely they live in environments around pH 7. In consequence it causes the protein to denature. Hydrogen peroxide is an oxidising agent which has the ability to damage cells and allows the formation of other free groups. Although the sum of H2O2 nowadays in honey is really low it is still effectual as an antimicrobic agent. Its sum varies depending on the type of honey. The low H2O activity of the honey inhibits bacteriums through the consequence of osmosis which causes plasmolysis of the cell which finally cause cell decease ( El-Sabbahy, 2009 ) .
Other mostly uncharacterized substances are besides present in some honeys which are thought to hold antibacterial effects. Some compounds that have been identified include syringic acid, methyl syringate, 3,4,5-trimethoxybenzoic acid, 2-hydroxy-3-phenylpropionic acid and the flavonoid pinocembrin ( honey scientific study ) . Manuka honey from New Zealand has been found to hold significant degrees of non-peroxide antibacterial activity and unidentified phytochemical constituent ( Peter Charles Molan-Honey as a topical agent for ) . It contains acidic and phenolic substances, the most dominant being 2-hydroxy-3-phenylpropionic acid as demonstrated in gas chromatography by Tan et Al ( 1989 ) . Australian honeys were identified besides of holding volatile substances which contribute to honey ‘s antibacterial activity ( Molan, 2001 ) .
Recently there are researches that have shown that some honeys have an immunologic activity. Honey stimulates the proliferation of peripheral blood B-lymphocytes and T-lymphocytes in cell civilization ( Abuharfeil et. al. , 1999 ) . It besides stimulates monocytes in cell civilization to let go of cytokines, tumour mortification factor-alpha, interleukin-1 and IL-6 ( Tonks et. al. , 2001 ) .
A survey conducted by Sam El-Sabbahy ( 2009 ) on the antibacterial effects of different honeys on different bacterial species showed that all honeys exhibited antibacterial activity which increased in increasing concentrations. Honeies used were Manuka honey, forest honey, organic flowered honey, Royale honey, and Turkish trefoil honey. It showed besides that different honeys act otherwise on the same micro-organism because the composing of each honey is different. Furthermore, it revealed that the same honey Acts of the Apostless otherwise on different micro-organisms.
Robson et Al ( 2007 ) concluded in their survey that the usage of honey in wound direction following ENT surgery is effectual. It enhances rapid healing of the lesion.
Another survey by Talal et Al ( 2008 ) found that honey is more effectual in killing both planktonic and biofilm-grown signifiers of bacteriums compared with the rate of bacteriacide by antibiotics normally used against bacteriums.
Research Question: “ What is non yet known about the subject? ”
The usage of honey in speed uping lesion healing and its antibacterial consequence have been proven and investigated exhaustively. However, small is known as to its antimicrobic consequence in instances of caput and cervix infective diseases peculiarly in sinonasal country.
General inquiry:
The survey aims to find the in-vitro antibacterial consequence of two available organic flowered honey in Toril, Davao City on the common infective bacteriums in ague and chronic sinusitis.
Sub inquiries:
1. Does the locally available organic flowered honey possess an antibacterial consequence o the common infective bacteriums in ague and chronic sinusitis?
2. Which among the two types of organic flowered honey has a better antibacterial belongings against the common pathogens in ague and chronic sinusitis.
3. What degree of dilution of honey does it still possess its antibacterial authority to the common pathogens in ague and chronic sinusitis?
4. What common infective bacteriums in ague and chronic sinusitis are susceptible to honey?
Significance of the Study: “ What will the health care be if the replies to the research inquiry will be known? ”
Because of the outgrowth of antibiotic- opposition bacteria the involvement in research is now geared towards the usage of other intervention options such as alternate medicative therapies that are effectual, readily available and low-cost. This survey will look into and research the in-vitro antibacterial consequence of locally produced organic flowered honey on the common bacterial pathogens in ague and chronic sinusitis.
Aims: “ What will the survey make? ”
General:
To find if the locally available honey in Toril, Davao City has an antibacterial consequence on the common bacterial pathogens in ague and chronic sinusitis.
Specific:
1. To find whether the available two different types of organic flowered honey has an antibacterial belongings.
2. To compare which of the two different organic flowered honey has a better antibacterial activity.
3. To find up to what degree of dilution of honey will it still have an antibacterial consequence
4. To find what common pathogens in ague and chronic sinusitis are susceptible to honey.
Methodology
Study Design: Experimental Survey
Puting: Southern Philippines Medical Center
Materials:
Honey samples
Two different samples of honey ( Type A and B ) of organic type coming from different flowered beginnings will be used in the survey. These will be obtained from a apiarist in Toril, Davao City.
Bacterial species
There are five different species of bacterium which will be used in the survey. These are the common bacterial pathogens doing ague and chronic rhinosinusitis. The pure civilizations of these bacteriums will be obtained from the research lab of Southern Philippines Medical Center. The five bacterial species are: Strep. Pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Staph. aureus, and Bacteroides spp.
Methods
The undermentioned processs are adapted from the old survey by EL Sabbahy ( 2009 ) .
Beginnings of Isolates
The civilizations of the five different species will be used to individually inoculate the five different food agar plates utilizing an inoculant cringle to streak the bacteriums onto each home base
Culture Incubation and Storage Condition
The five different bacterial species which will be used in the survey will be incubated at room temperatures for up to 48 hour. The bacterial civilizations will so be stored at 4EsC.
Detection of Antibacterial Activity
Culture readying
Five cosmopolitan bottles incorporating 9 ml each of alimentary stock will be inoculated individually with the bacterial species utilizing a cringle. The alimentary stock solutions will so be incubated at 37 EsC for up to 48 hours.
Honey readying
There will be five different concentrations which will be prepared from each of the two types of organic flowered honey. The honey sample will be diluted with unfertile H2O to 0 % ( negative control ) , 25 % , 50 % , 75 % , and 100 % .
10 milliliter from each dilution from the two types of organic flowered honey will be poured into five separate flasks and will be covered with foil. These were stored at room temperature out of direct sunshine.
Disc diffusion check
Five sets of five Nutrient Agar home base will be set ( will be done in triplicates ) , each agar home base in every set will be incubated individually with the bacterial species, by pipetting 100 uml of each bacteriums straight onto the agar surface o each home base of every set. Using the spread home base technique, the bacterium samples will so be spread across the surface utilizing a glass spreader.
The home bases will be left to dry for 15 proceedingss, while unfertile absorbent phonograph record will be placed into each honey flask. The absorptive phonograph record will be left in honey for 10 proceedingss to absorb the honey. An absorptive phonograph record from each honey will be placed on every agar home base in each set.
The home bases inoculated by the five bacterial species will so be incubated at 37EsC for up to 48 hours.
Each experiment will be done in triplicate to hold an accurate consequence.
Detection of antibacterial activity and minimal inhibitory concentration
After 48 hours of incubation, the bacterial growing suppression will be determined by the ocular verification of a zone of suppression, which is a clear country environing the absorbent phonograph record. There is positive suppression ( + ) if there will be a seeable clear pealing around the absorbent phonograph record, and there is negative suppression ( – ) if there will be no clear ring. The sum of suppression will be recorded by mensurating the diameter of the zone of suppression, in millimetres ( millimeter ) , this will be measured utilizing a swayer. The measuring will include the diameter of the absorbent phonograph record.
Datas Analysis
There will be three statistical methods that will be used in the analysis of the consequences. These include mean ( x ) , Standard Deviation ( SD ) and the Percentage of Coefficient of Variation ( % CV ) .
The mean will be used to find the mean suppression of each honey at each concentration of suppression. The Percentage Coefficient of Variation will be used to find the sum of fluctuation of each consequence.
To find the sum of antibacterial activity for each honey and to bring forth a more precise Minimum Inhibitory Concentration measurement dose response curve will be generated by plotting the zone of inhibition- diameter squared ( mm square ) against the degree of concentration. This will be carried out on each type of honey which will be used I this probe for each of the species of bacteriums used.
Datas Analysis Plan
The dummy tabular arraies below will be used as a usher for informations analysis.
Table 1.Bacterial Inhibition at 100 % concentration of honey
Strep pneumoniae
H. grippe
Moraxella catarrhalis
Staph. Aureus
Bacteroides spp
Honey A
R1
R2
R3
Honey B
R1
R2
R3
( + ) positive suppression ( – ) No seeable suppression
Table 2. Disc Diffusion Assay for the minimal Inhibitory Concentration on each bacterial pathogen. The Average ( ten ) . Standard Deviation ( SD ) and Percentage Coefficient of Variation ( % CV ) .
