Research ArticleAntibacterial Activity of Honey against Methicillin-ResistantStaphylococcus aureus A Laboratory-Based Experimental Study

Mohammedaman Mama 1 Teklu Teshome2 and Jafer Detamo3

1Madda Walabu University Goba Referral Hospital Department of Medical Laboratory Science Goba Ethiopia2Arba Minch University College of Medicine and Health Sciences Department of Biomedical Sciences Arba Minch Ethiopia3Arba Minch University College of Medicine and Health Sciences Department of Pharmacy Arba Minch Ethiopia

Correspondence should be addressed to Mohammedaman Mama muheekoogmailcom

Received 25 December 2018 Accepted 20 March 2019 Published 3 April 2019

Academic Editor Joseph Falkinham

Copyright copy 2019 Mohammedaman Mama et al (is is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in anymedium provided the original work isproperly cited

Background Antimicrobial drug resistance is one of the serious issues this world is facing nowadays and increased cost ofsearching for effective antimicrobial agents and the decreased rate of new drug discovery have made the situation increasinglyworrisome Objective (e aim of this study is to determine in vitro antibacterial activity of honey against methicillin-resistantStaphylococcus aureus isolates from wound infection Methods An experimental study was conducted from May to November2017 Methicillin resistance was detected using cefoxitin (30 μg) and oxacillin (1 μg) antibiotic discs Different concentrations ofhoney (25ndash100 vv) were tested against each type of clinical isolates obtained from wound infection A preliminary sensitivitytest was done to all types of honey by using disk diffusion while minimum inhibitory concentration and minimum bactericidalconcentration were determined for the most potent honey by the broth dilution technique All statistical analysis was performedby using Statistical Package for the Social Sciences version 20 Results In this study 36 bacterial isolates were recovered from 50specimens showing an isolation rate of 72 (e predominant bacteria isolated from the infected wounds were Staphylococcusaureus (15 417) Among identified Staphylococcus aureus methicillin resistance accounts for 10 isolates (278) All isolatesshowed a high frequency of resistance to tetracycline Four collected honey varieties exhibited antibacterial activity while thestrongest inhibitory activity was demonstrated by honey-2 at 75 vv (e mean MIC and MBC of honey-2 ranged from 938 to375 vv Conclusions Tested honey has both a bacteriostatic and bactericidal activity Among the tested honey ldquohoney-2rdquo hadhigh antibacterial potency than others

1 Introduction

Inadequate wound management compounded with sec-ondary infection is still remaining a major public healthproblem in developing countries (erefore wound man-agement has remained as research focus so far Because ofincreased drug resistance the interest in using alternativetherapies and natural remedies in wound management hasrapidly increased [1 2]

Despite the enormous advance in health care madeduring the last half-century infectious diseases still accountfor 25 of mortality worldwide and 45 in low-incomecountries Among these pathogenic and antibiotic-resistantbacteria pose a very serious threat to public health which

makes them the main cause of mortality and morbidity inhospitals and community [3ndash7]

Anti-infective drugs (antimicrobial agents) are criticallyimportant in reducing the global burden of infectious dis-eases [8 9] (e occurrence of drug-resistant microorgan-isms diminished the development of antibiotics and fewpharmaceutical companies remain active in this area posinga big challenge in this world [10] Hence the failure of theseantibiotics has resulted for a man to search for more effectivesources of natural products from plants and others [11 12]

(e antibacterial activity of honey was first recognized in1892 however it has a limited use in modern medicine dueto lack of scientific support [13] Honey is the nectar col-lected from flowers by bees It contains 15 to 20 water

HindawiInternational Journal of MicrobiologyVolume 2019 Article ID 7686130 9 pageshttpsdoiorg10115520197686130

and 80 to 85 sugar (e remainder of the honey is madeup of proteins enzymes and nonessential amino acids[14ndash16] Several properties of honey like enzymes are re-sponsible for its bactericidal effect and wound healingGlucose oxidase which changes glucose to gluconolactoneand then to hydrogen peroxide is one of the enzymes foundin honey (e release of H2O2 is slow and continuous for aconstant antibacterial effect successfully eliminating mi-croorganisms but dilute enough not to damage host tissue[17ndash19]

Honey having acidic pH of 32ndash45 is used to inhibitmany pathogenic organisms and increased wound healingprocess through epithelization [17 20 21] Honey is also oneof the supersaturated solutions that inhibit bacterial growthprimarily due to this high osmolarity [22]

(e World Health Organization (WHO) has describedalternative medicines as a cheap way to achieve total healthcare coverage of the worldrsquos population and has encouragedthe rational use of plant-based alternative medicines bymember states [23] In Ethiopia about 80 of the totalpopulation relies on traditional remedies as a primary sourceof health care [24] Hence the purpose of this study was toevaluate antibacterial activity (bacteriostatic and bactericidaleffects) of honey against MRSA isolates from an infectedwound in Gamo Gofa Zone South Ethiopia so that theywould have been recommended as therapeutic agents afterpharmaceutical standardization and clinical trials

2 Materials and Methods

21 Study Design Area and Period An experimental studydesign was conducted at Arba Minch University MedicalMicrobiology and Parasitology Laboratory from May 2017to November 2017

22 Sample Size Determination and Sampling Technique(e sample size was determined using sample size de-termination for estimation of single population proportionformula Taking 97 prevalence of multiple drug resistance(MDR) isolates from previous study [25] 95 confidenceinterval (z 196) and 5 marginal error (d 005) theinitial sample size is

n zprop2( 1113857

2 lowastP(1minusP)

d2 (196)2 lowast 097lowast 003

(005)2asymp 45 (1)

Finally by considering a 10 ( asymp5 subjects) nonresponserate the final sample size was determined as n + 5 asymp 50

Regarding sampling technique all isolated MRSA wereincluded in the experiment

23 Wound Sampling Procedure Open and clinically in-fected wound swabs were aseptically obtained after thewound was cleansed with sterile normal saline and collectedby trained nurses (e specimen was collected on sterilecotton swab by rotating with sufficient pressure(e sampleswere transported to the laboratory after collection usingAmies transport media

24 Culture and Identification Swabs collected werestreaked on blood agar and mannitol salt agar (Oxoid) byusing a sterile inoculation loop (e plates were incubated at35ndash37degC for 24ndash48 hours Preliminary identification ofbacteria was based on colony characteristics of the organ-isms such as hemolysis on blood agar changes in physicalappearance in differential media and enzyme activities ofthe organisms Isolates were identified based on their gramreaction catalase and coagulase test results

25 Antibacterial Susceptibility Testing Susceptibility testingwas performed by KirbyndashBauer disk diffusion techniqueaccording to criteria set by Clinical Laboratory StandardInstitute (CLSI) 2016 (e inoculums were prepared andsuspended in sterile normal saline(e density of suspensionwas determined by comparison with opacity standard onMcFarland 05 barium sulphate solution (e test organismwas uniformly seeded over the MuellerndashHinton agar(Oxoid) surface and exposed to the concentration gradientof the antibiotic followed by incubation at 37degC for 16ndash18 hours Diameters of the zone of inhibition around thediscs were measured to the nearest millimeter using a rulerand classified as sensitive intermediate and resistantaccording to the standardized table supplied by CLSI 2016(e antibiotics tested were ciprofloxacin (5 μg) gentamicin(10 μg) tetracycline (30 μg) co-trimoxazole (25 μg) chlor-amphenicol (30 μg) amikacin (30 μg) clindamycin (10 μg)erythromycin (15 μg) and vancomycin (30 μg) (ese anti-microbials were selected based on the availability and pre-scription frequency of these drugs in the study area

26 Test Organisms All screened methicillin-resistant Saureus from the wound was used Methicillin-resistant Saureus was identified phenotypically based on its resistance tooxacillin (1 μg) and cefoxitin (30 μg) by the disc diffusionmethod performed on modified MullerndashHinton agar (OxoidBasingstoke UK) Based on the CLSI 2016 guideline the zoneof inhibition is interpreted and grouped into methicillin-sensitive and methicillin-resistant S aureus [18]

27 Honey Sample Four kinds of honey were harvestedfrom beekeepers of the Gamo Gofa zone using purposivesampling technique Honey was collected in sterile screwedcupsculture bottle Each honey sample was first filtered witha sterile meshgauze to remove debris and then streaked onblood agar plate to check sterility and stored at 2ndash8degC untilused

28 Preparation of Honey Solutions Hundred percent purehoney (100 vv) was obtained after filtered using sterilegauze To get 75 honey solutions (vv) 075ml of honeywas diluted in 025ml sterilized distilled water Further serialdilutions of 05ml of each and 025ml of honey and 075mlof sterile distilled water were added to obtain 50 and 25honey solutions (vv) respectively

2 International Journal of Microbiology

29 Susceptibility Testing ofHoney Susceptibility testing wasperformed by KirbyndashBauer disk diffusion techniqueaccording to criteria set by CLSI 2016 (e inoculums wereprepared by picking parts of similar test organisms with asterile wire loop and suspended in sterile normal saline (edensity of suspension was determined by comparison withopacity standard on McFarland 05 barium sulphate solu-tion A sterile swab was dipped into the suspension of theisolate squeezed free from excess fluid against the side of thetube and then spread over the agar plate (e test organismwas uniformly seeded over the MuellerndashHinton agar(Oxoid) surface and the plates were left on the bench for theexcess fluid being absorbed Using a sterile cork borer (6mmdiameter 4mm deep and about 2 cm apart) wells weremade in the agar medium Using a micropipette 50 microL ofhoney with the concentration of 75 50 and 25 wasadded to the wells in the plate (e plates were incubated at37degC for 24 h (e mean diameters of inhibition zones weremeasured in mm and the results were recorded A positivecontrol well was equally filled with vancomycin (30 microg)while sterile distilled water used as negative control (eexperiment was repeated 3times for each strain

210 Determination of Minimum Inhibitory ConcentrationMinimum inhibitory concentration (MIC) and minimumbactericidal concentration (MBC) of the antimicrobialagents were determined for each isolate by the tube dilutionmethod Briefly ten sterile test tubes were placed in the racklabeled each 1 through 8 Honey control tubes (HC) brothcontrol tube (BC) and growth control tube (GC) were usedas quality controls One milliliter of freshly prepared nu-trient broth was added to each tube sterilized and cooled(en one milliliter of undiluted honey solution 100 wasadded to test tube number 1 and HC with a sterile mi-cropipette and tips (en twofold serial dilution was per-formed by transferring 1ml undiluted honey into the secondtube with separate sterile micropipette and tips and vortexedfor homogenization After a through mixing 1ml wastransferred with another sterile micropipette from tube 2and tube 3(ese procedures continued until the eighth tubewith a dilution of 1 128 was reached and finally 1ml wastaken and discarded from tube 8 (e GC tube that receivedno honey and BC that received no bacterial inoculumsserved as growth control while the HC tube that received nobacterial inoculums served as a honey control Except for theHC tube each tube was inoculated with 1ml of the culture ofthe respective prepared organism (e whole procedureswere repeated for all the organisms tested to each of thehoney Tubes were then incubated at 37degC for 24 h andobserved by visual inspections for the presence and absenceof growth (turbidity) MIC was recorded as the lowestconcentration of honey that inhibited bacterial growth (novisible growth or turbidity)

211 Determination of Minimum Bactericidal ConcentrationTo determine the MBC incubated tubes showing no visiblesign of growthturbidity in MIC were subcultured ontosterile nutrient agar plates by the streak plate method and

incubated at 37degC for 24 h aerobically (e least concen-tration of honey that did not show growth of test organismswas considered as the MBC (en inoculated plates werescored as bactericidal if no growth bacteriostatic if there islight to moderate growth and no antibacterial activity ifthere is heavy growth

212 Data Quality Control Data quality was ensured atvarious activities of the study (preanalytical analytical andpostanalytical) by following prepared standard operatingprocedure (SOP) Culture media was prepared according tothe manufacturerrsquos instruction and the sterility was checkedby incubating representative of the batch at 35ndash37degC over-night and observing bacterial growth (ose batch of themedia showed growth was discarded A control strain of Saureus (ATCC-25923) was used to check the quality of themedia and potency of the antibiotics used for positivecontrols Pretesting of the questionnaires was done on about5 of the total respondents and the completed question-naires were checked and corrections were made on a dailybase

3 Results

31 Study Population and Patient Characteristics A total of50 samples collected from patients with clinical evidence ofwound infection (patients with complaints of dischargepain swelling foul smelling and chronic wound) from Mayto November 2017 (e subjects included 25 (50) malesand females respectively (e incidence of the wound washighest among the age group 16ndash30 years with 25 (50)followed by age groupge 15 years 11 (22) among the totalnumber of patients studied Housewives and students (15(30)) had the highest infection rate among the occupa-tional group followed by government employees (8 (16))(e sociodemographic characteristics of the participants aresummarized in (Table 1)

32 Prevalence of MRSA As stated in Figure 1 amongisolated S aureus (15 (417)) screened for methicillin re-sistance MRSA accounts 10 (667) while the remaining 5(333) were methicillin-sensitive S aureus (MSSA)

33 Antibiotic Susceptibility Pattern Methicillin-resistantclinical isolates were tested against selected 9 antibioticsthat prescribed in the area and recommended by the ClinicalLaboratory Standard Susceptibility of pathogens to testedantibiotics was varied Eighty percent of pathogens wereresistant to tetracycline followed by 40 co-trimoxazole and30 erythromycin However all isolates showed high sen-sitivity to vancomycin amikacin ciprofloxacin and gen-tamicin (Table 2)

34 Disk Diffusion Honey Sensitivity Test In this study fourdifferent varieties of honey labeled as honey-1 to -4 werecollected and tested for their antimicrobial potential fromArba Minch Province on ten methicillin -resistant S aureus

International Journal of Microbiology 3

Table 1 Wound infection and sociodemographic characteristics of the patients with wound infection at Arba Minch Hospital Arba MinchMayndashNovember 2017

Demographic characters Infected (no ()) Not infected (no ()) Total (no ())SexMale 17 (68) 8 25 (50)Female 16 (64) 9 25 (50)Total 33 (66) 17 (34) 50 (100)Age in yearsle15 5 6 1116ndash30 20 (80) 5 25 (50)31ndash44 4 4 845ndash59 4 2 6Total 33 (66) 17 (34) 50 (100)OccupationStudents 12 3 15 (30)Housewives 10 5 15 (30)Civil servants 6 2 8Self-employed 2 3 5Merchants 2 2 4Farmers 1 2 3Total 33 (66) 17 (34) 50 (100)Educational statusIlliterate 9 7 16 (32)Literate 24 (706) 10 34 (68)Total 33 (66) 17 (34) 50 (100)

MSSAMRSA

3330

6670

Figure 1 Prevalence of MRSA and MSSA among patients with infected wounds at Arba Minch Hospital Arba Minch MayndashNovember2017

Table 2 Antimicrobial susceptibility pattern of MSSA and MRSA from infected wound patients at Arba Minch Hospital Arba MinchSouth Ethiopia MayndashNovember 2017

Isolate Antimicrobial agents (no ())VA CLN ERY AK TET CHL CPR COT GEN

MSSA (n 5) S 5 (100) 5 (100) 4 (80) 5100 2 (30) 3 (60) 4 (80) 4 (80) 5 (100)R 0 0 1 (20) 0 3 (60) 2 (30) 1 (20) 1 (20) 0

MRSA (n 10) S 10 (100) 9 (90) 7 (70) 10100 2 (20) 8 (80) 9 (90) 6 (60) 9 (90)R 0 1 (10) 3 (30) 0 8 (80) 2 (20) 1 (10) 4 (40) 1 (10)

Total (n 15) S 15 (100) 14 (933) 11 (733) 15100 4 (267) 11 (733) 13 (867) 10 (667) 14 (933)R 0 1 (67) 4 (267) 0 11 (733) 4 (267) 2 (133) 5 (333) 1 (67)

VA vancomycin CLN clindamycin ERY erythromycin AK amikacin TET tetracycline CHL chloramphenicol CPR ciprooxacin COTco-trimoxazole GEN gentamicin

4 International Journal of Microbiology

(MRSA) clinical isolates All collected honey samples showan antibacterial effect at 100 vv while some of themshowed a bactericidal and bacteriostatic effect at 75 and 50vv In general zones of inhibition ranged from 0ndash39mmaccordingly honey-2 at 75 vv concentration showed thelargest average zone of inhibition and selected for furtherminimum inhibitory concentration and minimum bacteri-cidal concentration (Figures 2 and 3)

35 Determination of MIC (e MIC determination wasdone only for honey-2 because of the potent antibacterialeffect is shown on disk diffusion during the preliminarysensitivity assay (e mean MICs of the honey-2 sampleswith clinical isolates of MRSA are presented in Table 3 (emean MIC of MRSA isolates ranged from 938 to 375 vvwhile most of the isolates 6 (60) showedMIC at 1875 vvand the least 1 (10) at 375 vv

36 Determination of MBC As mentioned above the mostactive honey was further assayed to determine its minimuminhibitory concentration and minimum bactericidal con-centration against 10 MRSA isolates Consequently honey-2at 75 vv was selected (e MBC ranged from 938 to 375(Table 4 and Figure 4) Partial inhibition of 50 of the testMRSA was observed starting from 1875 vv and 100complete inhibition was observed at 375 vv of honeyHence the MBC value of 30 of tested microorganisms wasfound to be similar to the MIC value of tested organisms at1875 vv

4 Discussion

Wound infections have been a problem in the field ofmedicine for a long time and the problem complicated morerecently because of increased antimicrobial resistance (isis a problem too for public researchers clinicians and drugcompanies looking for effective drugs (erefore antimi-crobial resistance may increase complications and costsassociated with procedures and treatment that leads to acontinued search for new agents [26]

S aureus is a Gram-positive bacterium which is a majorpathogen implicated in skin infections such as impetigofuruncles boils sties pustules burns and woundsAntibiotic-resistant strains of S aureus are the major causeof infections especially in a hospital setting [27] Strains of Saureus that were fully sensitive to penicillin now developedresistance to methicillin and other latest ones resort anti-biotics too [28]

In this study a total of 50 patients suffering from woundinfections were included of which male and female eachaccounts 25 (50) respectively (e incidence of woundinfection was relatively more common in males (68) thanin females (64) (is is in agreement with studies per-formed in different parts of Ethiopia like Bahirdar [29]Addis Ababa [30] and Gondar [31] Nigeria [32 33] andIndia [34] (is slight differences might be explained by thefact that traditionally in this country mainly males are

involved in occupations such as farming constructionworks transportation and industry works where the likelyexposure to trauma is common

Staphylococcus aureus has been known to acquire re-sistance to most antibiotics including the penicillinase-resistant ones like methicillin A study carried out in USA[35] found an incidence of 206MRSA and 10 in [36] and217 in [37] which are more lower when compared withthat in the current study (667) in contrast to this higherincidences of 45 and 582 MRSA have been documentedby Eagye et al [38] and Keith et al [39] As well higherincidence of (634) was reported in China [40] which issimilar to the current study We found that all the MRSAstrains were (100) sensitive to vancomycin and amikacinfollowed by (90) gentamicin and ciprofloxacin re-spectively(is finding could have relevant clinical use in theantibiotic policy guidelines for hospitals

Honey antimicrobial properties will vary depending onthe type of honey geographical location and flower fromwhich the final product is derived [41] Hence the presentstudy aimed to test the antimicrobial activity of honeyagainst MRSA isolates from wound infection

Hence in the present study all tested honey samplesshow an antibacterial effect against clinical isolates duringdisk diffusion technique Especially it showed completeinhibitory effect with a clear zone of inhibition against thetested organism at 100 vv(e bactericidal concentrationsof honey against MRSA in our study were between 50 and100 (is concentration was higher than the findings ofother researchers [19 42 43]

Tested honey sample (honey-2) was found to have bothbacteriostatic and bactericidal properties with differentconcentrations ranging from 938ndash375 vv Growth re-tardation and complete inhibition on 70 of the test or-ganisms were observed at a concentration of 1875 vv ofhoney

(e MIC and MBC values in this study indicated thattested honey (honey-2) has potential bactericidal and bac-teriostatic activities against multidrug-resistant clinicalisolates of MRSA bacteria (is was similar to other studiesconducted elsewhere [42 44 45]

(e percentage by volume of honey to completelyprevent the growth of MRSA was in the range of 1875ndash375 vv In contrary to this a study conducted in Ethiopiahas shown that the percentage by volume of honey tocompletely prevent growth of S aureus to be 65 vv [42]which is lower concentration than our result Another studybyWillix has also found that the (vv) of Manuka honey tocompletely prevent growth for S aureus was 18 [46] (isdifference might be due to the difference in the species ofbees and the differences in the test methods used and testorganisms or might be due to the variation in antimicrobialactivities of honey in different geographical locations inpreparing their valuable honey (nectars and pollens)

Even though tested honey shows antibacterial effectmany studies have demonstrated that not all honey sampleshave the same degree of antibacterial activity (erefore thesensitivity of MRSA isolates cannot be compared using theresults from different studies as the honey used in the

International Journal of Microbiology 5

studies may have had widely diiexclering antimicrobial activ-ities centerefore the MIC and MBC values determined withtheMRSA strains in this study indicate that there is no much

diiexclerence in sensitivity (eiexclectiveness of honey to inhibitgrowth or to kill the bacteria) to honey Hence honey haspotential in the decontamination of wounds colonized

0

MRS

A-1

MRS

A-2

MRS

A-3

MRS

A-4

MRS

A-5

MRS

A-6

MRS

A-7

MRS

A-8

MRS

A-9

MRS

A-1

0

5

10

15

20

25

30

35

40

45

Hon

ey-2

zone

of i

nhib

ition

agai

nst i

sola

tes

25 vv50 vv75 vv

Figure 2 Susceptibility of MRSA to honey-2 at 25 vv 50 vv and 75 vv collected from Arba Minch Province Arba MinchMayndashNovember 2017

(a) (b)

(c) (d)

Figure 3 Disk diiexclusion susceptibility ofMRSA to honey-2 at 25 vv 50 vv 75 vv and 100 vv collected fromArbaMinch ProvinceArba Minch MayndashNovember 2017

6 International Journal of Microbiology

antibiotic-resistant strains of bacteria like MRSA (issupports the existing local traditional practice of usinghoney to treat wound infections [47 48]

5 Conclusions

Honey had antimicrobial properties (bacteriostatic andbactericidal activities) against MRSA organisms tested (eantibacterial potency of ldquohoney-2rdquo on the test organism ofMRSA isolates was highly effective with MIC and MBCranged from 938ndash375 vv

Abbreviations

ATCC American Type Culture CollectionBC Broth controlCLSI Clinical Laboratory Standard InstituteGC Growth controlHC Honey controlMBC Minimum bactericidal concentrationMDR Multiple drug resistanceMIC Minimum inhibitory concentrationMRSA Methicillin-resistant Staphylococcus aureus

Table 3 MIC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 164 1128 MIC ( vv)MRSA-1 minus minus minus + + + + + 1875MRSA-2 minus minus minus minus + + + + 938MRSA-3 minus minus minus + + + + + 1875MRSA-4 minus minus minus minus + + + + 938MRSA-5 minus minus minus minus + + + + 938MRSA-6 minus minus minus + + + + + 1875MRSA-7 minus minus minus + + + + + 1875MRSA-8 minus minus + + + + + + 375MRSA-9 minus minus minus + + + + + 1875MRSA-10 minus minus minus + + + + + 1875minus no growth (bactericidal) + growth

(a) (b)

Figure 4 Minimum bactericidal determination against MRSA isolates from Arba Minch Province Arba Minch MayndashNovember 2017

Table 4 MBC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 MBC ( vv)MRSA-1 minus minus + ++ +++ +++ 375MRSA-2 minus minus minus + ++ +++ 1875MRSA-3 minus minus + ++ +++ +++ 375MRSA-4 minus minus minus + ++ +++ 1875MRSA-5 minus minus minus + ++ +++ 1875MRSA-6 minus minus + ++ ++ +++ 375MRSA-7 minus minus minus + ++ +++ 1875MRSA-8 minus minus + ++ +++ +++ 375MRSA-9 minus minus + ++ +++ +++ 375MRSA-10 minus minus minus + ++ +++ 1875minus no growth (bactericidal) + light growth ++ moderate growth (bacteriostatic) +++ heavy growth (no antibacterial potential)

International Journal of Microbiology 7

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

29 Susceptibility Testing ofHoney Susceptibility testing wasperformed by KirbyndashBauer disk diffusion techniqueaccording to criteria set by CLSI 2016 (e inoculums wereprepared by picking parts of similar test organisms with asterile wire loop and suspended in sterile normal saline (edensity of suspension was determined by comparison withopacity standard on McFarland 05 barium sulphate solu-tion A sterile swab was dipped into the suspension of theisolate squeezed free from excess fluid against the side of thetube and then spread over the agar plate (e test organismwas uniformly seeded over the MuellerndashHinton agar(Oxoid) surface and the plates were left on the bench for theexcess fluid being absorbed Using a sterile cork borer (6mmdiameter 4mm deep and about 2 cm apart) wells weremade in the agar medium Using a micropipette 50 microL ofhoney with the concentration of 75 50 and 25 wasadded to the wells in the plate (e plates were incubated at37degC for 24 h (e mean diameters of inhibition zones weremeasured in mm and the results were recorded A positivecontrol well was equally filled with vancomycin (30 microg)while sterile distilled water used as negative control (eexperiment was repeated 3times for each strain

210 Determination of Minimum Inhibitory ConcentrationMinimum inhibitory concentration (MIC) and minimumbactericidal concentration (MBC) of the antimicrobialagents were determined for each isolate by the tube dilutionmethod Briefly ten sterile test tubes were placed in the racklabeled each 1 through 8 Honey control tubes (HC) brothcontrol tube (BC) and growth control tube (GC) were usedas quality controls One milliliter of freshly prepared nu-trient broth was added to each tube sterilized and cooled(en one milliliter of undiluted honey solution 100 wasadded to test tube number 1 and HC with a sterile mi-cropipette and tips (en twofold serial dilution was per-formed by transferring 1ml undiluted honey into the secondtube with separate sterile micropipette and tips and vortexedfor homogenization After a through mixing 1ml wastransferred with another sterile micropipette from tube 2and tube 3(ese procedures continued until the eighth tubewith a dilution of 1 128 was reached and finally 1ml wastaken and discarded from tube 8 (e GC tube that receivedno honey and BC that received no bacterial inoculumsserved as growth control while the HC tube that received nobacterial inoculums served as a honey control Except for theHC tube each tube was inoculated with 1ml of the culture ofthe respective prepared organism (e whole procedureswere repeated for all the organisms tested to each of thehoney Tubes were then incubated at 37degC for 24 h andobserved by visual inspections for the presence and absenceof growth (turbidity) MIC was recorded as the lowestconcentration of honey that inhibited bacterial growth (novisible growth or turbidity)

211 Determination of Minimum Bactericidal ConcentrationTo determine the MBC incubated tubes showing no visiblesign of growthturbidity in MIC were subcultured ontosterile nutrient agar plates by the streak plate method and

incubated at 37degC for 24 h aerobically (e least concen-tration of honey that did not show growth of test organismswas considered as the MBC (en inoculated plates werescored as bactericidal if no growth bacteriostatic if there islight to moderate growth and no antibacterial activity ifthere is heavy growth

212 Data Quality Control Data quality was ensured atvarious activities of the study (preanalytical analytical andpostanalytical) by following prepared standard operatingprocedure (SOP) Culture media was prepared according tothe manufacturerrsquos instruction and the sterility was checkedby incubating representative of the batch at 35ndash37degC over-night and observing bacterial growth (ose batch of themedia showed growth was discarded A control strain of Saureus (ATCC-25923) was used to check the quality of themedia and potency of the antibiotics used for positivecontrols Pretesting of the questionnaires was done on about5 of the total respondents and the completed question-naires were checked and corrections were made on a dailybase

3 Results

31 Study Population and Patient Characteristics A total of50 samples collected from patients with clinical evidence ofwound infection (patients with complaints of dischargepain swelling foul smelling and chronic wound) from Mayto November 2017 (e subjects included 25 (50) malesand females respectively (e incidence of the wound washighest among the age group 16ndash30 years with 25 (50)followed by age groupge 15 years 11 (22) among the totalnumber of patients studied Housewives and students (15(30)) had the highest infection rate among the occupa-tional group followed by government employees (8 (16))(e sociodemographic characteristics of the participants aresummarized in (Table 1)

32 Prevalence of MRSA As stated in Figure 1 amongisolated S aureus (15 (417)) screened for methicillin re-sistance MRSA accounts 10 (667) while the remaining 5(333) were methicillin-sensitive S aureus (MSSA)

33 Antibiotic Susceptibility Pattern Methicillin-resistantclinical isolates were tested against selected 9 antibioticsthat prescribed in the area and recommended by the ClinicalLaboratory Standard Susceptibility of pathogens to testedantibiotics was varied Eighty percent of pathogens wereresistant to tetracycline followed by 40 co-trimoxazole and30 erythromycin However all isolates showed high sen-sitivity to vancomycin amikacin ciprofloxacin and gen-tamicin (Table 2)

34 Disk Diffusion Honey Sensitivity Test In this study fourdifferent varieties of honey labeled as honey-1 to -4 werecollected and tested for their antimicrobial potential fromArba Minch Province on ten methicillin -resistant S aureus

International Journal of Microbiology 3

Table 1 Wound infection and sociodemographic characteristics of the patients with wound infection at Arba Minch Hospital Arba MinchMayndashNovember 2017

Demographic characters Infected (no ()) Not infected (no ()) Total (no ())SexMale 17 (68) 8 25 (50)Female 16 (64) 9 25 (50)Total 33 (66) 17 (34) 50 (100)Age in yearsle15 5 6 1116ndash30 20 (80) 5 25 (50)31ndash44 4 4 845ndash59 4 2 6Total 33 (66) 17 (34) 50 (100)OccupationStudents 12 3 15 (30)Housewives 10 5 15 (30)Civil servants 6 2 8Self-employed 2 3 5Merchants 2 2 4Farmers 1 2 3Total 33 (66) 17 (34) 50 (100)Educational statusIlliterate 9 7 16 (32)Literate 24 (706) 10 34 (68)Total 33 (66) 17 (34) 50 (100)

MSSAMRSA

3330

6670

Figure 1 Prevalence of MRSA and MSSA among patients with infected wounds at Arba Minch Hospital Arba Minch MayndashNovember2017

Table 2 Antimicrobial susceptibility pattern of MSSA and MRSA from infected wound patients at Arba Minch Hospital Arba MinchSouth Ethiopia MayndashNovember 2017

Isolate Antimicrobial agents (no ())VA CLN ERY AK TET CHL CPR COT GEN

MSSA (n 5) S 5 (100) 5 (100) 4 (80) 5100 2 (30) 3 (60) 4 (80) 4 (80) 5 (100)R 0 0 1 (20) 0 3 (60) 2 (30) 1 (20) 1 (20) 0

MRSA (n 10) S 10 (100) 9 (90) 7 (70) 10100 2 (20) 8 (80) 9 (90) 6 (60) 9 (90)R 0 1 (10) 3 (30) 0 8 (80) 2 (20) 1 (10) 4 (40) 1 (10)

Total (n 15) S 15 (100) 14 (933) 11 (733) 15100 4 (267) 11 (733) 13 (867) 10 (667) 14 (933)R 0 1 (67) 4 (267) 0 11 (733) 4 (267) 2 (133) 5 (333) 1 (67)

VA vancomycin CLN clindamycin ERY erythromycin AK amikacin TET tetracycline CHL chloramphenicol CPR ciprooxacin COTco-trimoxazole GEN gentamicin

4 International Journal of Microbiology

(MRSA) clinical isolates All collected honey samples showan antibacterial effect at 100 vv while some of themshowed a bactericidal and bacteriostatic effect at 75 and 50vv In general zones of inhibition ranged from 0ndash39mmaccordingly honey-2 at 75 vv concentration showed thelargest average zone of inhibition and selected for furtherminimum inhibitory concentration and minimum bacteri-cidal concentration (Figures 2 and 3)

35 Determination of MIC (e MIC determination wasdone only for honey-2 because of the potent antibacterialeffect is shown on disk diffusion during the preliminarysensitivity assay (e mean MICs of the honey-2 sampleswith clinical isolates of MRSA are presented in Table 3 (emean MIC of MRSA isolates ranged from 938 to 375 vvwhile most of the isolates 6 (60) showedMIC at 1875 vvand the least 1 (10) at 375 vv

36 Determination of MBC As mentioned above the mostactive honey was further assayed to determine its minimuminhibitory concentration and minimum bactericidal con-centration against 10 MRSA isolates Consequently honey-2at 75 vv was selected (e MBC ranged from 938 to 375(Table 4 and Figure 4) Partial inhibition of 50 of the testMRSA was observed starting from 1875 vv and 100complete inhibition was observed at 375 vv of honeyHence the MBC value of 30 of tested microorganisms wasfound to be similar to the MIC value of tested organisms at1875 vv

4 Discussion

Wound infections have been a problem in the field ofmedicine for a long time and the problem complicated morerecently because of increased antimicrobial resistance (isis a problem too for public researchers clinicians and drugcompanies looking for effective drugs (erefore antimi-crobial resistance may increase complications and costsassociated with procedures and treatment that leads to acontinued search for new agents [26]

S aureus is a Gram-positive bacterium which is a majorpathogen implicated in skin infections such as impetigofuruncles boils sties pustules burns and woundsAntibiotic-resistant strains of S aureus are the major causeof infections especially in a hospital setting [27] Strains of Saureus that were fully sensitive to penicillin now developedresistance to methicillin and other latest ones resort anti-biotics too [28]

In this study a total of 50 patients suffering from woundinfections were included of which male and female eachaccounts 25 (50) respectively (e incidence of woundinfection was relatively more common in males (68) thanin females (64) (is is in agreement with studies per-formed in different parts of Ethiopia like Bahirdar [29]Addis Ababa [30] and Gondar [31] Nigeria [32 33] andIndia [34] (is slight differences might be explained by thefact that traditionally in this country mainly males are

involved in occupations such as farming constructionworks transportation and industry works where the likelyexposure to trauma is common

Staphylococcus aureus has been known to acquire re-sistance to most antibiotics including the penicillinase-resistant ones like methicillin A study carried out in USA[35] found an incidence of 206MRSA and 10 in [36] and217 in [37] which are more lower when compared withthat in the current study (667) in contrast to this higherincidences of 45 and 582 MRSA have been documentedby Eagye et al [38] and Keith et al [39] As well higherincidence of (634) was reported in China [40] which issimilar to the current study We found that all the MRSAstrains were (100) sensitive to vancomycin and amikacinfollowed by (90) gentamicin and ciprofloxacin re-spectively(is finding could have relevant clinical use in theantibiotic policy guidelines for hospitals

Honey antimicrobial properties will vary depending onthe type of honey geographical location and flower fromwhich the final product is derived [41] Hence the presentstudy aimed to test the antimicrobial activity of honeyagainst MRSA isolates from wound infection

Hence in the present study all tested honey samplesshow an antibacterial effect against clinical isolates duringdisk diffusion technique Especially it showed completeinhibitory effect with a clear zone of inhibition against thetested organism at 100 vv(e bactericidal concentrationsof honey against MRSA in our study were between 50 and100 (is concentration was higher than the findings ofother researchers [19 42 43]

Tested honey sample (honey-2) was found to have bothbacteriostatic and bactericidal properties with differentconcentrations ranging from 938ndash375 vv Growth re-tardation and complete inhibition on 70 of the test or-ganisms were observed at a concentration of 1875 vv ofhoney

(e MIC and MBC values in this study indicated thattested honey (honey-2) has potential bactericidal and bac-teriostatic activities against multidrug-resistant clinicalisolates of MRSA bacteria (is was similar to other studiesconducted elsewhere [42 44 45]

(e percentage by volume of honey to completelyprevent the growth of MRSA was in the range of 1875ndash375 vv In contrary to this a study conducted in Ethiopiahas shown that the percentage by volume of honey tocompletely prevent growth of S aureus to be 65 vv [42]which is lower concentration than our result Another studybyWillix has also found that the (vv) of Manuka honey tocompletely prevent growth for S aureus was 18 [46] (isdifference might be due to the difference in the species ofbees and the differences in the test methods used and testorganisms or might be due to the variation in antimicrobialactivities of honey in different geographical locations inpreparing their valuable honey (nectars and pollens)

Even though tested honey shows antibacterial effectmany studies have demonstrated that not all honey sampleshave the same degree of antibacterial activity (erefore thesensitivity of MRSA isolates cannot be compared using theresults from different studies as the honey used in the

International Journal of Microbiology 5

studies may have had widely diiexclering antimicrobial activ-ities centerefore the MIC and MBC values determined withtheMRSA strains in this study indicate that there is no much

diiexclerence in sensitivity (eiexclectiveness of honey to inhibitgrowth or to kill the bacteria) to honey Hence honey haspotential in the decontamination of wounds colonized

0

MRS

A-1

MRS

A-2

MRS

A-3

MRS

A-4

MRS

A-5

MRS

A-6

MRS

A-7

MRS

A-8

MRS

A-9

MRS

A-1

0

5

10

15

20

25

30

35

40

45

Hon

ey-2

zone

of i

nhib

ition

agai

nst i

sola

tes

25 vv50 vv75 vv

Figure 2 Susceptibility of MRSA to honey-2 at 25 vv 50 vv and 75 vv collected from Arba Minch Province Arba MinchMayndashNovember 2017

(a) (b)

(c) (d)

Figure 3 Disk diiexclusion susceptibility ofMRSA to honey-2 at 25 vv 50 vv 75 vv and 100 vv collected fromArbaMinch ProvinceArba Minch MayndashNovember 2017

6 International Journal of Microbiology

antibiotic-resistant strains of bacteria like MRSA (issupports the existing local traditional practice of usinghoney to treat wound infections [47 48]

5 Conclusions

Honey had antimicrobial properties (bacteriostatic andbactericidal activities) against MRSA organisms tested (eantibacterial potency of ldquohoney-2rdquo on the test organism ofMRSA isolates was highly effective with MIC and MBCranged from 938ndash375 vv

Abbreviations

ATCC American Type Culture CollectionBC Broth controlCLSI Clinical Laboratory Standard InstituteGC Growth controlHC Honey controlMBC Minimum bactericidal concentrationMDR Multiple drug resistanceMIC Minimum inhibitory concentrationMRSA Methicillin-resistant Staphylococcus aureus

Table 3 MIC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 164 1128 MIC ( vv)MRSA-1 minus minus minus + + + + + 1875MRSA-2 minus minus minus minus + + + + 938MRSA-3 minus minus minus + + + + + 1875MRSA-4 minus minus minus minus + + + + 938MRSA-5 minus minus minus minus + + + + 938MRSA-6 minus minus minus + + + + + 1875MRSA-7 minus minus minus + + + + + 1875MRSA-8 minus minus + + + + + + 375MRSA-9 minus minus minus + + + + + 1875MRSA-10 minus minus minus + + + + + 1875minus no growth (bactericidal) + growth

(a) (b)

Figure 4 Minimum bactericidal determination against MRSA isolates from Arba Minch Province Arba Minch MayndashNovember 2017

Table 4 MBC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 MBC ( vv)MRSA-1 minus minus + ++ +++ +++ 375MRSA-2 minus minus minus + ++ +++ 1875MRSA-3 minus minus + ++ +++ +++ 375MRSA-4 minus minus minus + ++ +++ 1875MRSA-5 minus minus minus + ++ +++ 1875MRSA-6 minus minus + ++ ++ +++ 375MRSA-7 minus minus minus + ++ +++ 1875MRSA-8 minus minus + ++ +++ +++ 375MRSA-9 minus minus + ++ +++ +++ 375MRSA-10 minus minus minus + ++ +++ 1875minus no growth (bactericidal) + light growth ++ moderate growth (bacteriostatic) +++ heavy growth (no antibacterial potential)

International Journal of Microbiology 7

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

Table 1 Wound infection and sociodemographic characteristics of the patients with wound infection at Arba Minch Hospital Arba MinchMayndashNovember 2017

Demographic characters Infected (no ()) Not infected (no ()) Total (no ())SexMale 17 (68) 8 25 (50)Female 16 (64) 9 25 (50)Total 33 (66) 17 (34) 50 (100)Age in yearsle15 5 6 1116ndash30 20 (80) 5 25 (50)31ndash44 4 4 845ndash59 4 2 6Total 33 (66) 17 (34) 50 (100)OccupationStudents 12 3 15 (30)Housewives 10 5 15 (30)Civil servants 6 2 8Self-employed 2 3 5Merchants 2 2 4Farmers 1 2 3Total 33 (66) 17 (34) 50 (100)Educational statusIlliterate 9 7 16 (32)Literate 24 (706) 10 34 (68)Total 33 (66) 17 (34) 50 (100)

MSSAMRSA

3330

6670

Figure 1 Prevalence of MRSA and MSSA among patients with infected wounds at Arba Minch Hospital Arba Minch MayndashNovember2017

Table 2 Antimicrobial susceptibility pattern of MSSA and MRSA from infected wound patients at Arba Minch Hospital Arba MinchSouth Ethiopia MayndashNovember 2017

Isolate Antimicrobial agents (no ())VA CLN ERY AK TET CHL CPR COT GEN

MSSA (n 5) S 5 (100) 5 (100) 4 (80) 5100 2 (30) 3 (60) 4 (80) 4 (80) 5 (100)R 0 0 1 (20) 0 3 (60) 2 (30) 1 (20) 1 (20) 0

MRSA (n 10) S 10 (100) 9 (90) 7 (70) 10100 2 (20) 8 (80) 9 (90) 6 (60) 9 (90)R 0 1 (10) 3 (30) 0 8 (80) 2 (20) 1 (10) 4 (40) 1 (10)

Total (n 15) S 15 (100) 14 (933) 11 (733) 15100 4 (267) 11 (733) 13 (867) 10 (667) 14 (933)R 0 1 (67) 4 (267) 0 11 (733) 4 (267) 2 (133) 5 (333) 1 (67)

VA vancomycin CLN clindamycin ERY erythromycin AK amikacin TET tetracycline CHL chloramphenicol CPR ciprooxacin COTco-trimoxazole GEN gentamicin

4 International Journal of Microbiology

(MRSA) clinical isolates All collected honey samples showan antibacterial effect at 100 vv while some of themshowed a bactericidal and bacteriostatic effect at 75 and 50vv In general zones of inhibition ranged from 0ndash39mmaccordingly honey-2 at 75 vv concentration showed thelargest average zone of inhibition and selected for furtherminimum inhibitory concentration and minimum bacteri-cidal concentration (Figures 2 and 3)

35 Determination of MIC (e MIC determination wasdone only for honey-2 because of the potent antibacterialeffect is shown on disk diffusion during the preliminarysensitivity assay (e mean MICs of the honey-2 sampleswith clinical isolates of MRSA are presented in Table 3 (emean MIC of MRSA isolates ranged from 938 to 375 vvwhile most of the isolates 6 (60) showedMIC at 1875 vvand the least 1 (10) at 375 vv

36 Determination of MBC As mentioned above the mostactive honey was further assayed to determine its minimuminhibitory concentration and minimum bactericidal con-centration against 10 MRSA isolates Consequently honey-2at 75 vv was selected (e MBC ranged from 938 to 375(Table 4 and Figure 4) Partial inhibition of 50 of the testMRSA was observed starting from 1875 vv and 100complete inhibition was observed at 375 vv of honeyHence the MBC value of 30 of tested microorganisms wasfound to be similar to the MIC value of tested organisms at1875 vv

4 Discussion

Wound infections have been a problem in the field ofmedicine for a long time and the problem complicated morerecently because of increased antimicrobial resistance (isis a problem too for public researchers clinicians and drugcompanies looking for effective drugs (erefore antimi-crobial resistance may increase complications and costsassociated with procedures and treatment that leads to acontinued search for new agents [26]

S aureus is a Gram-positive bacterium which is a majorpathogen implicated in skin infections such as impetigofuruncles boils sties pustules burns and woundsAntibiotic-resistant strains of S aureus are the major causeof infections especially in a hospital setting [27] Strains of Saureus that were fully sensitive to penicillin now developedresistance to methicillin and other latest ones resort anti-biotics too [28]

In this study a total of 50 patients suffering from woundinfections were included of which male and female eachaccounts 25 (50) respectively (e incidence of woundinfection was relatively more common in males (68) thanin females (64) (is is in agreement with studies per-formed in different parts of Ethiopia like Bahirdar [29]Addis Ababa [30] and Gondar [31] Nigeria [32 33] andIndia [34] (is slight differences might be explained by thefact that traditionally in this country mainly males are

involved in occupations such as farming constructionworks transportation and industry works where the likelyexposure to trauma is common

Staphylococcus aureus has been known to acquire re-sistance to most antibiotics including the penicillinase-resistant ones like methicillin A study carried out in USA[35] found an incidence of 206MRSA and 10 in [36] and217 in [37] which are more lower when compared withthat in the current study (667) in contrast to this higherincidences of 45 and 582 MRSA have been documentedby Eagye et al [38] and Keith et al [39] As well higherincidence of (634) was reported in China [40] which issimilar to the current study We found that all the MRSAstrains were (100) sensitive to vancomycin and amikacinfollowed by (90) gentamicin and ciprofloxacin re-spectively(is finding could have relevant clinical use in theantibiotic policy guidelines for hospitals

Honey antimicrobial properties will vary depending onthe type of honey geographical location and flower fromwhich the final product is derived [41] Hence the presentstudy aimed to test the antimicrobial activity of honeyagainst MRSA isolates from wound infection

Hence in the present study all tested honey samplesshow an antibacterial effect against clinical isolates duringdisk diffusion technique Especially it showed completeinhibitory effect with a clear zone of inhibition against thetested organism at 100 vv(e bactericidal concentrationsof honey against MRSA in our study were between 50 and100 (is concentration was higher than the findings ofother researchers [19 42 43]

Tested honey sample (honey-2) was found to have bothbacteriostatic and bactericidal properties with differentconcentrations ranging from 938ndash375 vv Growth re-tardation and complete inhibition on 70 of the test or-ganisms were observed at a concentration of 1875 vv ofhoney

(e MIC and MBC values in this study indicated thattested honey (honey-2) has potential bactericidal and bac-teriostatic activities against multidrug-resistant clinicalisolates of MRSA bacteria (is was similar to other studiesconducted elsewhere [42 44 45]

(e percentage by volume of honey to completelyprevent the growth of MRSA was in the range of 1875ndash375 vv In contrary to this a study conducted in Ethiopiahas shown that the percentage by volume of honey tocompletely prevent growth of S aureus to be 65 vv [42]which is lower concentration than our result Another studybyWillix has also found that the (vv) of Manuka honey tocompletely prevent growth for S aureus was 18 [46] (isdifference might be due to the difference in the species ofbees and the differences in the test methods used and testorganisms or might be due to the variation in antimicrobialactivities of honey in different geographical locations inpreparing their valuable honey (nectars and pollens)

Even though tested honey shows antibacterial effectmany studies have demonstrated that not all honey sampleshave the same degree of antibacterial activity (erefore thesensitivity of MRSA isolates cannot be compared using theresults from different studies as the honey used in the

International Journal of Microbiology 5

studies may have had widely diiexclering antimicrobial activ-ities centerefore the MIC and MBC values determined withtheMRSA strains in this study indicate that there is no much

diiexclerence in sensitivity (eiexclectiveness of honey to inhibitgrowth or to kill the bacteria) to honey Hence honey haspotential in the decontamination of wounds colonized

0

MRS

A-1

MRS

A-2

MRS

A-3

MRS

A-4

MRS

A-5

MRS

A-6

MRS

A-7

MRS

A-8

MRS

A-9

MRS

A-1

0

5

10

15

20

25

30

35

40

45

Hon

ey-2

zone

of i

nhib

ition

agai

nst i

sola

tes

25 vv50 vv75 vv

Figure 2 Susceptibility of MRSA to honey-2 at 25 vv 50 vv and 75 vv collected from Arba Minch Province Arba MinchMayndashNovember 2017

(a) (b)

(c) (d)

Figure 3 Disk diiexclusion susceptibility ofMRSA to honey-2 at 25 vv 50 vv 75 vv and 100 vv collected fromArbaMinch ProvinceArba Minch MayndashNovember 2017

6 International Journal of Microbiology

antibiotic-resistant strains of bacteria like MRSA (issupports the existing local traditional practice of usinghoney to treat wound infections [47 48]

5 Conclusions

Honey had antimicrobial properties (bacteriostatic andbactericidal activities) against MRSA organisms tested (eantibacterial potency of ldquohoney-2rdquo on the test organism ofMRSA isolates was highly effective with MIC and MBCranged from 938ndash375 vv

Abbreviations

ATCC American Type Culture CollectionBC Broth controlCLSI Clinical Laboratory Standard InstituteGC Growth controlHC Honey controlMBC Minimum bactericidal concentrationMDR Multiple drug resistanceMIC Minimum inhibitory concentrationMRSA Methicillin-resistant Staphylococcus aureus

Table 3 MIC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 164 1128 MIC ( vv)MRSA-1 minus minus minus + + + + + 1875MRSA-2 minus minus minus minus + + + + 938MRSA-3 minus minus minus + + + + + 1875MRSA-4 minus minus minus minus + + + + 938MRSA-5 minus minus minus minus + + + + 938MRSA-6 minus minus minus + + + + + 1875MRSA-7 minus minus minus + + + + + 1875MRSA-8 minus minus + + + + + + 375MRSA-9 minus minus minus + + + + + 1875MRSA-10 minus minus minus + + + + + 1875minus no growth (bactericidal) + growth

(a) (b)

Figure 4 Minimum bactericidal determination against MRSA isolates from Arba Minch Province Arba Minch MayndashNovember 2017

Table 4 MBC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 MBC ( vv)MRSA-1 minus minus + ++ +++ +++ 375MRSA-2 minus minus minus + ++ +++ 1875MRSA-3 minus minus + ++ +++ +++ 375MRSA-4 minus minus minus + ++ +++ 1875MRSA-5 minus minus minus + ++ +++ 1875MRSA-6 minus minus + ++ ++ +++ 375MRSA-7 minus minus minus + ++ +++ 1875MRSA-8 minus minus + ++ +++ +++ 375MRSA-9 minus minus + ++ +++ +++ 375MRSA-10 minus minus minus + ++ +++ 1875minus no growth (bactericidal) + light growth ++ moderate growth (bacteriostatic) +++ heavy growth (no antibacterial potential)

International Journal of Microbiology 7

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

(MRSA) clinical isolates All collected honey samples showan antibacterial effect at 100 vv while some of themshowed a bactericidal and bacteriostatic effect at 75 and 50vv In general zones of inhibition ranged from 0ndash39mmaccordingly honey-2 at 75 vv concentration showed thelargest average zone of inhibition and selected for furtherminimum inhibitory concentration and minimum bacteri-cidal concentration (Figures 2 and 3)

35 Determination of MIC (e MIC determination wasdone only for honey-2 because of the potent antibacterialeffect is shown on disk diffusion during the preliminarysensitivity assay (e mean MICs of the honey-2 sampleswith clinical isolates of MRSA are presented in Table 3 (emean MIC of MRSA isolates ranged from 938 to 375 vvwhile most of the isolates 6 (60) showedMIC at 1875 vvand the least 1 (10) at 375 vv

36 Determination of MBC As mentioned above the mostactive honey was further assayed to determine its minimuminhibitory concentration and minimum bactericidal con-centration against 10 MRSA isolates Consequently honey-2at 75 vv was selected (e MBC ranged from 938 to 375(Table 4 and Figure 4) Partial inhibition of 50 of the testMRSA was observed starting from 1875 vv and 100complete inhibition was observed at 375 vv of honeyHence the MBC value of 30 of tested microorganisms wasfound to be similar to the MIC value of tested organisms at1875 vv

4 Discussion

Wound infections have been a problem in the field ofmedicine for a long time and the problem complicated morerecently because of increased antimicrobial resistance (isis a problem too for public researchers clinicians and drugcompanies looking for effective drugs (erefore antimi-crobial resistance may increase complications and costsassociated with procedures and treatment that leads to acontinued search for new agents [26]

S aureus is a Gram-positive bacterium which is a majorpathogen implicated in skin infections such as impetigofuruncles boils sties pustules burns and woundsAntibiotic-resistant strains of S aureus are the major causeof infections especially in a hospital setting [27] Strains of Saureus that were fully sensitive to penicillin now developedresistance to methicillin and other latest ones resort anti-biotics too [28]

In this study a total of 50 patients suffering from woundinfections were included of which male and female eachaccounts 25 (50) respectively (e incidence of woundinfection was relatively more common in males (68) thanin females (64) (is is in agreement with studies per-formed in different parts of Ethiopia like Bahirdar [29]Addis Ababa [30] and Gondar [31] Nigeria [32 33] andIndia [34] (is slight differences might be explained by thefact that traditionally in this country mainly males are

involved in occupations such as farming constructionworks transportation and industry works where the likelyexposure to trauma is common

Staphylococcus aureus has been known to acquire re-sistance to most antibiotics including the penicillinase-resistant ones like methicillin A study carried out in USA[35] found an incidence of 206MRSA and 10 in [36] and217 in [37] which are more lower when compared withthat in the current study (667) in contrast to this higherincidences of 45 and 582 MRSA have been documentedby Eagye et al [38] and Keith et al [39] As well higherincidence of (634) was reported in China [40] which issimilar to the current study We found that all the MRSAstrains were (100) sensitive to vancomycin and amikacinfollowed by (90) gentamicin and ciprofloxacin re-spectively(is finding could have relevant clinical use in theantibiotic policy guidelines for hospitals

Honey antimicrobial properties will vary depending onthe type of honey geographical location and flower fromwhich the final product is derived [41] Hence the presentstudy aimed to test the antimicrobial activity of honeyagainst MRSA isolates from wound infection

Hence in the present study all tested honey samplesshow an antibacterial effect against clinical isolates duringdisk diffusion technique Especially it showed completeinhibitory effect with a clear zone of inhibition against thetested organism at 100 vv(e bactericidal concentrationsof honey against MRSA in our study were between 50 and100 (is concentration was higher than the findings ofother researchers [19 42 43]

Tested honey sample (honey-2) was found to have bothbacteriostatic and bactericidal properties with differentconcentrations ranging from 938ndash375 vv Growth re-tardation and complete inhibition on 70 of the test or-ganisms were observed at a concentration of 1875 vv ofhoney

(e MIC and MBC values in this study indicated thattested honey (honey-2) has potential bactericidal and bac-teriostatic activities against multidrug-resistant clinicalisolates of MRSA bacteria (is was similar to other studiesconducted elsewhere [42 44 45]

(e percentage by volume of honey to completelyprevent the growth of MRSA was in the range of 1875ndash375 vv In contrary to this a study conducted in Ethiopiahas shown that the percentage by volume of honey tocompletely prevent growth of S aureus to be 65 vv [42]which is lower concentration than our result Another studybyWillix has also found that the (vv) of Manuka honey tocompletely prevent growth for S aureus was 18 [46] (isdifference might be due to the difference in the species ofbees and the differences in the test methods used and testorganisms or might be due to the variation in antimicrobialactivities of honey in different geographical locations inpreparing their valuable honey (nectars and pollens)

Even though tested honey shows antibacterial effectmany studies have demonstrated that not all honey sampleshave the same degree of antibacterial activity (erefore thesensitivity of MRSA isolates cannot be compared using theresults from different studies as the honey used in the

International Journal of Microbiology 5

studies may have had widely diiexclering antimicrobial activ-ities centerefore the MIC and MBC values determined withtheMRSA strains in this study indicate that there is no much

diiexclerence in sensitivity (eiexclectiveness of honey to inhibitgrowth or to kill the bacteria) to honey Hence honey haspotential in the decontamination of wounds colonized

0

MRS

A-1

MRS

A-2

MRS

A-3

MRS

A-4

MRS

A-5

MRS

A-6

MRS

A-7

MRS

A-8

MRS

A-9

MRS

A-1

0

5

10

15

20

25

30

35

40

45

Hon

ey-2

zone

of i

nhib

ition

agai

nst i

sola

tes

25 vv50 vv75 vv

Figure 2 Susceptibility of MRSA to honey-2 at 25 vv 50 vv and 75 vv collected from Arba Minch Province Arba MinchMayndashNovember 2017

(a) (b)

(c) (d)

Figure 3 Disk diiexclusion susceptibility ofMRSA to honey-2 at 25 vv 50 vv 75 vv and 100 vv collected fromArbaMinch ProvinceArba Minch MayndashNovember 2017

6 International Journal of Microbiology

antibiotic-resistant strains of bacteria like MRSA (issupports the existing local traditional practice of usinghoney to treat wound infections [47 48]

5 Conclusions

Honey had antimicrobial properties (bacteriostatic andbactericidal activities) against MRSA organisms tested (eantibacterial potency of ldquohoney-2rdquo on the test organism ofMRSA isolates was highly effective with MIC and MBCranged from 938ndash375 vv

Abbreviations

ATCC American Type Culture CollectionBC Broth controlCLSI Clinical Laboratory Standard InstituteGC Growth controlHC Honey controlMBC Minimum bactericidal concentrationMDR Multiple drug resistanceMIC Minimum inhibitory concentrationMRSA Methicillin-resistant Staphylococcus aureus

Table 3 MIC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 164 1128 MIC ( vv)MRSA-1 minus minus minus + + + + + 1875MRSA-2 minus minus minus minus + + + + 938MRSA-3 minus minus minus + + + + + 1875MRSA-4 minus minus minus minus + + + + 938MRSA-5 minus minus minus minus + + + + 938MRSA-6 minus minus minus + + + + + 1875MRSA-7 minus minus minus + + + + + 1875MRSA-8 minus minus + + + + + + 375MRSA-9 minus minus minus + + + + + 1875MRSA-10 minus minus minus + + + + + 1875minus no growth (bactericidal) + growth

(a) (b)

Figure 4 Minimum bactericidal determination against MRSA isolates from Arba Minch Province Arba Minch MayndashNovember 2017

Table 4 MBC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 MBC ( vv)MRSA-1 minus minus + ++ +++ +++ 375MRSA-2 minus minus minus + ++ +++ 1875MRSA-3 minus minus + ++ +++ +++ 375MRSA-4 minus minus minus + ++ +++ 1875MRSA-5 minus minus minus + ++ +++ 1875MRSA-6 minus minus + ++ ++ +++ 375MRSA-7 minus minus minus + ++ +++ 1875MRSA-8 minus minus + ++ +++ +++ 375MRSA-9 minus minus + ++ +++ +++ 375MRSA-10 minus minus minus + ++ +++ 1875minus no growth (bactericidal) + light growth ++ moderate growth (bacteriostatic) +++ heavy growth (no antibacterial potential)

International Journal of Microbiology 7

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

studies may have had widely diiexclering antimicrobial activ-ities centerefore the MIC and MBC values determined withtheMRSA strains in this study indicate that there is no much

diiexclerence in sensitivity (eiexclectiveness of honey to inhibitgrowth or to kill the bacteria) to honey Hence honey haspotential in the decontamination of wounds colonized

0

MRS

A-1

MRS

A-2

MRS

A-3

MRS

A-4

MRS

A-5

MRS

A-6

MRS

A-7

MRS

A-8

MRS

A-9

MRS

A-1

0

5

10

15

20

25

30

35

40

45

Hon

ey-2

zone

of i

nhib

ition

agai

nst i

sola

tes

25 vv50 vv75 vv

Figure 2 Susceptibility of MRSA to honey-2 at 25 vv 50 vv and 75 vv collected from Arba Minch Province Arba MinchMayndashNovember 2017

(a) (b)

(c) (d)

Figure 3 Disk diiexclusion susceptibility ofMRSA to honey-2 at 25 vv 50 vv 75 vv and 100 vv collected fromArbaMinch ProvinceArba Minch MayndashNovember 2017

6 International Journal of Microbiology

antibiotic-resistant strains of bacteria like MRSA (issupports the existing local traditional practice of usinghoney to treat wound infections [47 48]

5 Conclusions

Honey had antimicrobial properties (bacteriostatic andbactericidal activities) against MRSA organisms tested (eantibacterial potency of ldquohoney-2rdquo on the test organism ofMRSA isolates was highly effective with MIC and MBCranged from 938ndash375 vv

Abbreviations

ATCC American Type Culture CollectionBC Broth controlCLSI Clinical Laboratory Standard InstituteGC Growth controlHC Honey controlMBC Minimum bactericidal concentrationMDR Multiple drug resistanceMIC Minimum inhibitory concentrationMRSA Methicillin-resistant Staphylococcus aureus

Table 3 MIC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 164 1128 MIC ( vv)MRSA-1 minus minus minus + + + + + 1875MRSA-2 minus minus minus minus + + + + 938MRSA-3 minus minus minus + + + + + 1875MRSA-4 minus minus minus minus + + + + 938MRSA-5 minus minus minus minus + + + + 938MRSA-6 minus minus minus + + + + + 1875MRSA-7 minus minus minus + + + + + 1875MRSA-8 minus minus + + + + + + 375MRSA-9 minus minus minus + + + + + 1875MRSA-10 minus minus minus + + + + + 1875minus no growth (bactericidal) + growth

(a) (b)

Figure 4 Minimum bactericidal determination against MRSA isolates from Arba Minch Province Arba Minch MayndashNovember 2017

Table 4 MBC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 MBC ( vv)MRSA-1 minus minus + ++ +++ +++ 375MRSA-2 minus minus minus + ++ +++ 1875MRSA-3 minus minus + ++ +++ +++ 375MRSA-4 minus minus minus + ++ +++ 1875MRSA-5 minus minus minus + ++ +++ 1875MRSA-6 minus minus + ++ ++ +++ 375MRSA-7 minus minus minus + ++ +++ 1875MRSA-8 minus minus + ++ +++ +++ 375MRSA-9 minus minus + ++ +++ +++ 375MRSA-10 minus minus minus + ++ +++ 1875minus no growth (bactericidal) + light growth ++ moderate growth (bacteriostatic) +++ heavy growth (no antibacterial potential)

International Journal of Microbiology 7

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

antibiotic-resistant strains of bacteria like MRSA (issupports the existing local traditional practice of usinghoney to treat wound infections [47 48]

5 Conclusions

Honey had antimicrobial properties (bacteriostatic andbactericidal activities) against MRSA organisms tested (eantibacterial potency of ldquohoney-2rdquo on the test organism ofMRSA isolates was highly effective with MIC and MBCranged from 938ndash375 vv

Abbreviations

ATCC American Type Culture CollectionBC Broth controlCLSI Clinical Laboratory Standard InstituteGC Growth controlHC Honey controlMBC Minimum bactericidal concentrationMDR Multiple drug resistanceMIC Minimum inhibitory concentrationMRSA Methicillin-resistant Staphylococcus aureus

Table 3 MIC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 164 1128 MIC ( vv)MRSA-1 minus minus minus + + + + + 1875MRSA-2 minus minus minus minus + + + + 938MRSA-3 minus minus minus + + + + + 1875MRSA-4 minus minus minus minus + + + + 938MRSA-5 minus minus minus minus + + + + 938MRSA-6 minus minus minus + + + + + 1875MRSA-7 minus minus minus + + + + + 1875MRSA-8 minus minus + + + + + + 375MRSA-9 minus minus minus + + + + + 1875MRSA-10 minus minus minus + + + + + 1875minus no growth (bactericidal) + growth

(a) (b)

Figure 4 Minimum bactericidal determination against MRSA isolates from Arba Minch Province Arba Minch MayndashNovember 2017

Table 4 MBC ( vv) of honey-2 samples against MRSA isolates in Arba Minch Province Arba Minch MayndashNovember 2017

Honey dilutionTest bacteria Net (1) 12 14 18 116 132 MBC ( vv)MRSA-1 minus minus + ++ +++ +++ 375MRSA-2 minus minus minus + ++ +++ 1875MRSA-3 minus minus + ++ +++ +++ 375MRSA-4 minus minus minus + ++ +++ 1875MRSA-5 minus minus minus + ++ +++ 1875MRSA-6 minus minus + ++ ++ +++ 375MRSA-7 minus minus minus + ++ +++ 1875MRSA-8 minus minus + ++ +++ +++ 375MRSA-9 minus minus + ++ +++ +++ 375MRSA-10 minus minus minus + ++ +++ 1875minus no growth (bactericidal) + light growth ++ moderate growth (bacteriostatic) +++ heavy growth (no antibacterial potential)

International Journal of Microbiology 7

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

MSSA Methicillin-susceptible Staphylococcus aureusSOP Standard operating procedureWHO World Health Organization

Data Availability

(e data used to support the findings of this study are in-cluded within the article

Conflicts of Interest

(e authors declare that they have no conflicts of interest

Acknowledgments

Our heartfelt thanks go to ArbaMinch University College ofMedicine and Health Sciences for funding the researchVery special thanks are due to ethical review boards of ArbaMinch University for giving ethical clearance We are alsoindebted to the study participants and data collectors

References

[1] H Maghsoudi F Salehi M K Khosrowshahi M BaghaeiM Nasirzadeh and R Shams ldquoComparison between topicalhoney and mafenide acetate in treatment of burn woundsrdquoAnnals of Burns and Fire Disasters vol 24 no 3 pp 132ndash1372011

[2] M K Tan D S H Adli M A Tumiran M A Abdulla andK M Yusoff ldquo(e efficacy of gelam honey dressing towardsexcisional wound healingrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 8059326 pages 2012

[3] A Osho and O Bello ldquoAntimicrobial effect of honey pro-duced by on some common human pathogensrdquoAsian Journalof Experimental Biological Science vol 1 pp 875ndash880 2010

[4] J Davies ldquoInactivation of antibiotics and the dissemination ofresistance genesrdquo Science vol 264 no 5157 pp 375ndash3821994

[5] L Pieters and A J Vlietinck ldquoBioguided isolation of phar-macologically active plant components still a valuablestrategy for the finding of new lead compoundsrdquo Journal ofEthnopharmacology vol 100 no 1-2 pp 57ndash60 2005

[6] M J Balunas and A D Kinghorn ldquoDrug discovery frommedicinal plantsrdquo Life Sciences vol 78 no 5 pp 431ndash4412005

[7] P Kwakman L De Boer C Ruyter-Spira et al ldquoMedical-grade honey enriched with antimicrobial peptides has en-hanced activity against antibiotic-resistant pathogensrdquo Eu-ropean Journal of Clinical Microbiology amp Infectious Diseasesvol 30 no 2 pp 251ndash257 2011

[8] P Shears ldquoAntimicrobial resistance in the tropicsrdquo TropicalDoctor vol 30 no 2 pp 114ndash116 2000

[9] A Mulu B Tessema and F Derbie ldquoIn vitro assessment ofthe antimicrobial potential of honey on common humanpathogensrdquo Ethiopian Journal of Health Development vol 18no 2 pp 107ndash111 2005

[10] V Bansal B Medhi and P Pandhi ldquoHoneyndasha remedyrediscovered and its therapeutic utilityrdquo Kathmandu Uni-versity Medical Journal vol 3 no 3 pp 305ndash309 2005

[11] F Omoya and F Akharaiyi ldquoMixture of honey and gingerextract for antibacterial assessment on some clinical isolatesrdquo

International Journal of Pharmaceutical and Biomedical Re-search vol 2 no 1 pp 39ndash47 2011

[12] B Dixon ldquoBacteria canrsquot resist honeyrdquo Lancet InfectiousDiseases vol 3 no 2 p 116 2003

[13] D Mohapatra V (akur and S Brar ldquoAntibacterial efficacyof raw and processed honeyrdquo Biotechnology Research In-ternational vol 2011 Article ID 917505 6 pages 2011

[14] A Henriques S Jackson R Cooper and N Burton ldquoFreeradical production and quenching in honeys with woundhealing potentialrdquo Journal of Antimicrobial Chemotherapyvol 58 no 4 pp 773ndash777 2006

[15] A Sharp ldquoBeneficial effects of honey dressings in woundmanagementrdquoNursing Standard vol 24 no 7 pp 66ndash74 2009

[16] J White and L W Doner ldquoHoney composition and prop-ertiesrdquo Beekeeping in the United States Agriculture Handbookvol 335 pp 82ndash91 1980

[17] P B Olaitan O E Adeleke and I O Ola ldquoHoney a reservoirfor microorganisms and an inhibitory agent for microbesrdquoAfrican Health Sciences vol 7 no 3 pp 159ndash165 2007

[18] P Lusby A Coombes and J Wilkinson ldquoHoney a potentagent for wound healingrdquo Journal of WOCN vol 29 no 6pp 295ndash300 2002

[19] P C Molan ldquo(e antibacterial activity of honeyrdquo Bee Worldvol 73 no 1 pp 5ndash28 1992

[20] B Pieper ldquoHoney-based dressings and wound carerdquo Journalof Wound Ostomy and Continence Nursing vol 36 no 6p 589 2009

[21] R White and P Molan ldquoA summary of published clinicalresearch on honey in wound managementrdquo in Honey AModern Wound Management Product R White R Cooperand P Molan Eds Wounds UK Publications Aberdeen UK2005

[22] P C Molan ldquo(e role of honey in the management ofwoundsrdquo Journal of Wound Care vol 8 no 8 pp 415ndash4181999

[23] X Zhang General Guidelines for Methodologies on Researchand Evaluation of Traditional Medicine World Health Or-ganization Geneva Switzerland 2000

[24] K D Kassaye A Amberbir B Getachew and Y MussemaldquoA historical overview of traditional medicine practices andpolicy in Ethiopiardquo Ethiopian Journal of Health Developmentvol 20 no 2 pp 127ndash134 2007

[25] M Mama A Abdissa and T Sewunet ldquoAntimicrobial sus-ceptibility pattern of bacterial isolates from wound infectionand their sensitivity to alternative topical agents at JimmaUniversity Specialized Hospital South-West Ethiopiardquo An-nals of Clinical Microbiology and Antimicrobials vol 13 no 1p 14 2014

[26] A M Sule L O A (anni S O A Odu and O OlusanyaldquoBacterial pathogens associated with infected wounds in OgunState University Teaching Hospital Sagamu Nigeriardquo AfricanJournal of Clinical and Experimental Microbiology vol 3no 1 pp 13ndash16 2002

[27] A B Mudey N Kesharwani G A Mudey R C GoyalA K Dawale and V V Wagh ldquoHealth status and personalhygiene among food handlers working at food establishmentaround a rural teaching hospital in Wardha District ofMaharashtra Indiardquo Global Journal of Health Science vol 2no 2 p 198 2010

[28] D Naik and A Teclu ldquoA study on antimicrobial susceptibilitypattern in clinical isolates of Staphylococcus aureus in EritreardquoPan African Medical Journal vol 3 no 1 2010

[29] F Biadglegne B Abera A Alem and B Anagaw ldquoBacterialisolates from wound infection and their antimicrobial

8 International Journal of Microbiology

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

susceptibility pattern in Felege Hiwot Referral Hospital NorthWest Ethiopiardquo Ethiopian Journal of Health Sciences vol 19no 3 pp 173ndash178 2009

[30] K Tekie K Surgical wound infection in Tikur Anbessa Hospitalwith special emphasis on Pseudomonas aeruginosa 2008httpetdaaueduetdspacebitstream12345678926211KASSAYE20TEKIEpdf

[31] A Gelaw Isolation of bacterial pathogens from patients withpostoperative surgical site infections and possible sources ofinfections at University of Gondar Hospital Northwest Ethiopia2011 httpetdaaueduetdspacebitstream12345678921391ASCHALEW20GELAWpdf

[32] C N Ohalete R K Obi and M C EmeaKoroha ldquoBacteri-ology of different wound infection and their antimicrobialsusceptibility patterns in Imo state NigeriardquoWorld Journal ofPharmaceutical Sciences vol 13 no 3 pp 1155ndash1172 2012

[33] O E Amoran A O Sogebi and O M Fatugase ldquoRates andrisk factors associated with surgical site infections in a tertiarycare center in South-Western Nigeriardquo International Journalof Tropical Disease amp Health vol 3 no 1 pp 25ndash36 2014

[34] N Goswami A P Goswami C Tripathi H Trivedi andT Patel ldquoAntibiotic sensitivity profile of bacterial pathogensin postoperative wound infections at a tertiary care hospital inGujarat Indiardquo Journal of Pharmacology and Pharmaco-therapeutics vol 2 no 3 pp 158ndash164 2011

[35] J A Weigelt B A Lipsky Y P Tabak K G Derby M Kimand V Gupta ldquoSugical site infections causative pathogensand associated outcomesrdquo American Journal of InfectionControl vol 38 no 2 pp 112ndash120 2010

[36] A Aggarwal S Khanna U Arora and P Devi ldquoCorrelationof β-lactamase productionmethicillin resistance and phagepattern of Staphylococcus aureusrdquo Indian Journal of MedicalSciences vol 55 no 5 pp 253ndash256 2001

[37] H Kownhar E M Shankar R Vignesh R Sekar V Veluand U A Rao ldquoHigh isolation rate of Staphylococcus aureusfrom surgical site infections in an Indian hospitalrdquo Journal ofAntimicrobial Chemotherapy vol 61 no 3 pp 758ndash7602008

[38] K J Eagye A Kim S Laohavaleeson J L Kuti andD P Nicolau ldquoSurgical site infections does inadequate an-tibiotic therapy affect patient outcomesrdquo Surgical Infectionsvol 10 no 4 pp 323ndash331 2009

[39] S K Keith D J Anderson R Sloane et al ldquo(e effect ofsurgical site infection on older operative patientsrdquo Journal ofthe American Geriatrics Society vol 57 no 1 pp 46ndash54 2009

[40] S-H Wang Z-L Sun Y-J Guo et al ldquoMeticillin-resistantStaphylococcus aureus isolated from foot ulcers in diabeticpatients in a Chinese care hospital risk factors for infectionand prevalencerdquo Journal of Medical Microbiology vol 59no 10 pp 1219ndash1224 2010

[41] O Sherlock A Dolan R Athman et al ldquoComparison of theantimicrobial activity of ulmo honey from Chile and manukahoney against methicillin-resistant Staphylococcus aureusEscherichia coli and Pseudomonas aeruginosardquo BMC Com-plementary and Alternative Medicine vol 10 no 1 p 472010

[42] M Ahmed S Sahile and C Subramanian ldquoEvaluation ofantibacterial potential of honey against some common humanpathogens in North Gondar Zone of Ethiopiardquo InternationalJournal of Pure and Applied Zoology vol 2 no 4 pp 286ndash2952014

[43] P C Molan and B Julie ldquoUsing honey dressings the practicalconsiderationsrdquo Nursing Times vol 96 no 49 pp 36-372000

[44] K L Allen G Hutchinson and P C Molan ldquo(e potentialfor using honey to treat wounds infected with MRSA andVRErdquo in Proceedings of First World Healing Congress Mel-bourne Australia September 2000

[45] A Getaneh Y Belyhun F Moges et al ldquoIn vitro assessmentof the antimicrobial efect of Ethiopian multi-flora honey onmethicillin resistant Staphylococcus aureusrdquo InternationalJournal of Current Research and Academic Review vol 5no 11 pp 64ndash72 2013

[46] D J Willix P C Molan and C G Harfoot ldquoA comparison ofthe sensitivity of wound-infecting species of bacteria to theantibacterial activity of manuka honey and other honeyrdquoJournal of Applied Bacteriology vol 73 no 5 pp 388ndash3941992

[47] N A AL-Haj E Amghalia M N Shamsudin R AbdullahR Mohamed and Z Sekaw ldquoAntibacterial activity of honeyagainst methicillin-resistant Staphylococcus aureusrdquo ResearchJournal of Biological Sciences vol 4 no 8 pp 943ndash947 2009

[48] NM George and K F Cutting ldquoAntibacterial Honey in vitroactivity against clinical isolates of MRSA VRE and othermulti-resistant Gram-negative organisms including Pseudo-monas aeruginosardquo Wounds vol 19 pp 231ndash236 2007

International Journal of Microbiology 9

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