2019 V0L.- 2 ISSU.- 1. INTERNATIONAL JOURNAL OF APPLIED SCIENCE AND RESEARCH ISSN IS :

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1 A STUDY OF BACTERIAL ISOLATES AMONG HEALTHY SUBJECTS IN UNIVERSITY ENVIRONMENTS: A CASE STUDY OF ADEKUNLE AJASIN UNIVERSITY, AKUNGBA-AKOKO, ONDO STATE Emmanuel Olumuyiwa Onifade,, Grace Adeoji, Stephen Olaide Aremu, 3 Department of Microbiology, Faculty of Science, Adekunle Ajain Univerity, P.M.B 00, Akungba- Akoko, Ondo State, Nigeria Department of Biological Science, Federal Univerity of Agriculture, P.M.B 373, Makurdi, Benue State, Nigeria Siberian State Medical Univerity, Mokovky Tract, Tomk, Ruian Federation Abtract Bacteria among other microorganim viz fungi, virue, protozoan and even ome metazoan are ubiquitou and a well urvive in different environment. Depite the fact that ome microorganim are beneficial while other are not, yet the diverity in habitat occupied by bacteria make it eaier for them to caue infection epecially whenever the environment i conducive a in the cae of opportunitic infection. The tudy wa undertaken to identify the bacteria aociated with hand palm and armpit of healthy ubject and alo to evaluate the antibiotic enitivity pattern of the bacteria. The tudy population conited of even male and four female healthy tudent of Adekunle Ajain Univerity Akungba-Akoko, Ondo State Nigeria between the age of eighteen (8) and thirty (30) year old. All the bacteria encountered were cultured on tryptone oya agar and eoin methylene blue agar, while only Gram-poitive cocci were cultured on mannitol alt agar. The bacterial iolate were identified baed on cultural, morphological and biochemical characteritic. Out of the ixteen bacteria obtained, 4(5%) were Staphylococcu aureu, while Streptococcu pecie and Staphylococcu pecie were 3(8.75%) each. Other bacteria identified were (.5%) of each Corynebacterium pecie and Klebiella pecie. In addition, Clotridium pecie and Bacillu pecie contituted (6.5%) each. Antibiotic uceptibility pattern of the bacteria were determined by dic diffuion method. All the bacterial iolate teted howed multiple antibiotic reitance. Keyword: Hand palm, armpit, bacterial iolate, reitance pattern, uceptibility pattern, Gram poitive, Gram negative, antibiotic reitance. Introduction Microorganim which include bacteria, fungi, virue, protozoan and ome metazoan are found in a wide of habitat. Although many microorganim are beneficial and neceary for well of human being. So, microbial activitie may caue undeirable coequence uch a food poilage and dieae, Precott et al. (008). Human and animal have abundant normal flora which i the mixture of microorganim regularly found at any anatomical ite that uually do not produce dieae which can be reident. Variou clae of organim uch a bacteria, virue, fungi,and protozoa caue dieae in man, Salle (00). The importance of microorganim in dieae wa not immediately obviou to people and it took many year for cientit to etablih the connection beetwen them and illnee, Precott et al.(008).under normal circumtance, microbe do not thrive, but the cratching of the urface of the kin provide oppourtunitie for microbe to infect the tiue.for any aetiological agent to urvive and pread, there mut be an environmentally favoured place to reide, Murray (990).Very few of the numerou microorganim to which human are expoed to produce dieae and the ability Copyright IJASR All right reerved Page

2 to initiate dieae i determined by an organim virulence and hot factor, Murray (990). The effect of infectiou dieae range from mild to evere or deadly depending on the phyiology of the hot and pathogen involved, Precott et al.(008). Invetigation from the previou tudie on microbiology on kin reveal Staphylococcu epidermidi a the dominant bacterial pecie found on hand. Other regular reident are Staphylococcu homini and other coagulae-negative taphylococci, followed by coryneform bacteria uch a propionibacteria, corynebacteria, dermabacteria, and micrococci, Hay (993); Lee et al. (994). Furthermore, armpit i uually accompanied by body odour a a reult of the normal functional of apocrine gland, one of the two type of gland that produce weat which reulting to the body odour, Alkerman et al. (005). Hence, the temporary reident kin flora often include noocomial bacteria and fungi, Hay, (993); Noble (993). Freh apocrine weat i odourle but it develop it characteritic mell when weat i broken by the hair and kin cell in the armpit and eaten by bacteria. It i actually the bacteria that make the mell by releaing a chemical called 3-methyl-- hexenoid acid, Alkerman et al. (005). The microorganim that are contantlypreent on body urface are commenal, thee pecie are life-long member of the body normal microbial community, but are not found everywhere.there are many area of the human body that remain axenic, and in the abence of dieae, are never colonized by microorganim. Reident microbiota typically colonize the urface of the kin, mucou membrane, digetive tract, upper repiratory ytem and dital portion of the urinogenital ytem; where they play major role in maintaining health and normal function, Tami and Port (009). The organim are adapted to the non-invaive mode of life defined by the limitation of the environment. If forcefully removed from the retriction of that environment and introduced into the bloodtream or tiue, thee organim may become pathogenic, Mandell et al. (005). Tranient microflora may attempt to colonize theome area of the body a do reident microbiota, Tami and Port (009). Tranient microorganim are alo extremely important to undertand. Thee include food borne micro organim and even oil borne micro organim that make their way into the human digetive tract and, depending upon the characteritic of the pecific organim involved, either ubtly or dramatically influence the overall health of the human ytem, Alkerman et al. (005). Infectiou dieae, alo known a communicable dieae, contagiou dieae or tranmiible dieae comprie clinically evident illne reulting from the infection, preence and growth of pathogenic biological agent in an individual hot organim, Abad et al. (994). A common but mot irritating body odour i armpit odour; it i o embarraing; therefore hand hygiene cannot be overemphaized ince it ha been conidered to be the mot important tool in noocomial infection control, Alkerman et al., (005); Nytrom (994). Colonization of health care worker' hand with Staphylococcu aureu ha been decribed to range between 0.5 and 78.3%. Up to 4,000,000 cell can be found per hand. The colonization rate with Staphylococcu aureu wa higher among doctor (36%) than among nure (8%), a wa the bacterial denity of Staphylococcu aureu on the hand ( and 5%, repectively, with more than,000 CFU per hand). The carrier rate may be up to 8% if the health care worker contact patient with an atopic dermatiti which i colonized by Staphylococcu aureu, William et al.(999); Methicilin-reitant Staphylococcu aureu (MRSA) ha been iolated from the hand of up to 6.9% of health care worker. Vancomycin reitance enterococcu (VRE) can be found on the hand of up to 4% of health care worker. However, every peron ha weat gland which reponible for perpiration which eventually allow bacteria to thrive at the armpit, Alkerman et al. (005). Copyright IJASR All right reerved Page

3 Hand carriage of pathogen uch a Staphylococcu aureu, methicilin reitance Staphylococcu aureu or Staphylococcu epidermidi ha repeatedly been aociated with different type of Noocomial Infection (NI). Hence, boil (or furuncle) i a kin dieae caued by inflammation of hair follicle, reulting in localized accumulation of pu and dead tiue, Alkerman et al. (005). The analyi of outbreak revealed that dermatiti on the hand of health care worker wa a rik factor for colonization or for inadequate hand hygiene, reulting in variou type of Noocomial Infection. The hand and glove of 44 health care worker were ampled after care of vancomycin reitance enterococcu poitive patient. Glove were vancomycin reitance enterococcu poitive for 7 of 44 healthcare worker, and hand were poitive for 5 of 44, even though they had worn glove, Tenorio et al. (00). One health care worker wa even vancomycin reitance enterococcu poitive on the hand although the culture from the glove wa negative; Staphylococcu aureu i the mot common gram-poitive bacterium cauing Ni, Tenorio et al. (00). It frequency among all pathogen in NI varie between.% and 7.%, Wagner et al. (997); Vo et al., (994). Methicillin reitance Staphylococcu aureu (MRSA) i increaing worldwide, Wagner et al. (997); Vo et al., (994); Sartor et al. (995). Staphylococcu aureu can urvive on hand for at leat 50 minute, wherea, about 90% of men and 70% of women have the bacteria that produce mulder odour in their armpit, Alkerman et al. (005). In addition, Staphylococcu i a genu of Gram-poitive, non-motile, pherical bacteria occuring in grape-like cluter. They belong to family of Staphylococcaceae, and are facultatively anaerobic, Catalae-poitive, Oxidae-negative, ferment glucoe and have teichoic acid in their cell wall, Precott et al. (008).Two common pecie of Staphylococcu that live in aociation with human are Staphylococcu epidermidi which live normally on the kin and mucou membrane and Staphylococcu aureu, which may occur normally at variou locale, but in a particular on the naal membrane, Kenneth (008). Neverthele, vancomycin reitance enterococcu urvive on hand or glove for up to 60 min. On inanimate urface, Staphylococcu aureu and methicillin reitance Staphylococcu aureu may urvive for 7 month, with wild train urviving longer than laboratory train. Vancomycin reitance enterococcu may urvive on urface for 4 month. The long urvival on urface, together with the relatively hort urvival on hand, ugget that contaminated urface may well be the ource of tranient colonization depite negative hand culture, Reboli et al. (989). Staphylococcu aureu i one of the major reitant pathogen; found on the mucou membrane and the human kin of around a third of the population, it i extremely adaptable to antibiotic preure. Methicillin wa then the antibiotic of choice, but ha ince been replaced by oxacillin due to ignificant kidney toxicity. CA-MRSA (Community-acquired MRSA) ha now emerged a an epidemic that i reponible for rapidly progreive, fatal dieae including necrotizing pneumonia, evere epi and necrotizing faciiti. Methicillin-reitant Staphylococcu aureu (MRSA) i the mot frequently identified antimicrobial drug-reitant pathogen in US hopital. The epidemiology of infection caued by MRSA i rapidly changing. Outbreak of community-aociated (CA)-MRSA infection have been reported in correctional facilitie, among athletic team, among military recruit, in newborn nurerie, and among men who have ex with men, Bartzoka et al. (983). Meanwhile, with regard to the mechanim of Antibiotic Reitance Staphylococci, reearch by Zukaite et al. (000); Albert and Bruce (008) reveal that hopital train of Staphylococcu aureu are uually reitant to a variety of different antibiotic. So, a few train are reitant to all clinically ueful antibiotic except vancomycin, and vancomycin-reitant train are increaingly-reported. Methicillin reitance i widepread and mot methicillin-reitant train are alo multiply reitant. Phagocytoi i the major mechanim for combating taphylococcal infection, Zukaite et al. (000); Albert and Bruce (008). Hence, the treatment of hopital acquired infection i often caued by Copyright IJASR All right reerved Page 3

4 antibiotic methicilin reitant train (MRSA) and can only be treated with vancomycin or an alternative. However, many of the community acquired (CA) Staphylococcal infection are now methicillin reitant, (Chopra et al., 005; Darren et al., 006). The control of Healthcare Aociated Infection (HCAI) repreent a major challenge to hopital and healthcare provider. Tranmiion of pathogen on the hand of healthcare worker i the mot common caue of cro infection, Damani (997); hygienic hand wah hould be performed in order to remove or detroy tranient microorganim and to ubtantially reduce reident micro-organim during time when urgical procedure are performed and before all aeptic procedure on the ward. Hence, what hould be ued for performing a hygienic hand wah i an approved antieptic detergent uch a 4 % Chlorhexidine gluconate or 7.5% Povidone iodine) (Laron, 995). Armpit odour i caued by the bacteria which reidence at the armpit the apocrine ecrete vicou fluid which i kind of milky from the hair follicle while the eccrine gland ecrete water fluid. The bacteria thrive more in the weat under armpit leading to the odour. Other caue of the odour can be due to the digetive product which accumulated in the body, Alkerman et al. (005). If you weat profuely try a olution of hydrogen peroxide (H O) 3% in concentration for wahing underarm; ue good deodorant and antiperpirant. Becaue, deodorant make the kin to be acidic and it diallow bacteria to thrive while anti-perpirant block the weat gland to produce weat kin floraare uually non-pathogenic are either commenal or mutualitic, Alkerman et al. (005). Thu, the effect of extracellular product obtained from culture upernatant of Staphylococcu aureu train product to a freh medium timulated growth already after hour of incubation, with an approximately two-fold increae in cell denity a compared to an unupplemented medium, probably by promoting an initiation of growth accompanied by a reduction of the initial lag phae, Bukharin et al. (000). Fibrinolyin i an enzyme derived from plama of bovine origin or extracted from culture of certain bacteria. It i ued locally only and excluively together with the enzyme deoxyribonucleae (extracted from bovine pancrea). Fibrinolyin and deoxyribonucleae both act a lytic enzyme. The combination i available a ointment containing BU (Biological Unit) fibrinolyin and 666 BU deoxyribonucleae per gram, Birk et al. (983). More alo, by catalyzing the hydrolyi of hyaluronan, a contituent of the intertitial barrier, hyaluronidae lower the vicoity of hyaluronan, thereby increaing tiue permeability. Other tudie ugget no contribution, or effect independent of enzyme activity, Coka et al. (00).Coagulae i an extracellular protein which bind to prothrombin in the hot to form a complex called taphylothrombin. The proteae activity characteritic of thrombin i activated in the complex, reulting in the converion of fibrinogen to fibrin. Coagulae i a traditional marker for identifying Staphylococcu aureu in the clinical microbiology laboratory, Darren et al. (006). With regard to taphylokinae;many train of Staphylococcu aureu expre a plaminogen activator called taphylokinae. Thi factor lye fibrin. The genetic determinant i aociated with lyogenic bacteriophage. A complex formed between taphylokinae and plaminogen activate plamin-like proteolytic activity which caue diolution of fibrin clot, Chopra et al. (005). Furthermore, Staphylococcu aureu can expre a toxin that pecifically act on polymorphonuclear leukocyte. Phagocytoi i an important defene againt taphylococcal infection o leukocidin hould be a virulence factor, Starr and Engleberg (006). Exotoxin in Staphylococcu aureu can expre everal different type of protein toxin which are probably reponible for ymptom during infection. Thoe which damage the membrane of cell were dicued above under invaion or lye erythrocyte, cauing hemolyi, but it i unlikely that haemolyi i a relevant determinant of virulence in vivo. Leukocidin caue membrane damage to leukocyte, Albert (008). Copyright IJASR All right reerved Page 4

5 Streptococcu i a genu of Gram-poitive, nonmotile, pherical bacteria occuring in chain. Mot pecie are aprophyte and ome are pathogenic. Many pathogenic pecie are haemolytic.they are chemoheterotrophic, meophilic, nonporing, Gram-poitive cocci, which are facultatively anaerobic and Catalae-negative, Precott et al.(008).like Staphylococu, there are many pecie of Streptococcu found normally in the human body, many are part of the kin and upper repiratory tract of human, Elmer (006). The family of Corynebacteriaceae, and the group contain aerobic and facultative anaerobe, catalae-poitive, traight to lightly curved rod, often with tapperedvend, Precott et al. (008). A genu of Gram-poitive, motly aerobic, nonmotile, rod like bacteria that frequetly bear club-haped welling. The genu contain the pecie Corynebacterium diphtheriae and non-diphtherial corynebacterium, Coyle et al. (990). Corynebacteria are non-porulating, Pleomorphic bacillio. They are chemoorganotrophic and exhibit a fermentation metabolim under certain condition; and are fatidiou organim growing on enrichment medium, Kenneth (008). Hand with artificial fingernail harbour gram-negative bacteria more often than thoe without. Health care worker have three opportunitie for the pot contamination treatment of hand viz: the ocial hand wah, the hygienic and, the hygienic hand diinfection which normally conit of the application of an alcohol-baed hand rub into dry hand without water, Hedderwick et al. (000). About 5% of all patient admitted to hopital develop a noocomial infection. Contaminated hand of health care peronnel are reported a a major route for the pread of noocomial infection Aitken et al. (00); Alfurayh et al. (000). Notwithtanding, another bacteria i Klebiella pecie which are gram negative rod haped bacteria which exhibit mucoid growth, large polyaccharide capule, and lack of motility, and they uually give poitive tet for lyine decarboxylae and citrate. Klebiella wa named after the German bacteriologit Edwin Kleb (834 93). Multiple-reitant Klebiella pneumoniae have been killed in vivo via intra peritoneal, intravenou or intranaal adminitration of phage in laboratory tet. While thi treatment ha been available for ome time, a greater danger of bacterial reitance exit to phage than to antibiotic, Cogen et al. (008). Reitance to phage may caue a bloom in the number of the microbe in environment a well a among human (if not obligate pathogenic). Thi i why phage therapy i only ued in conjunction with antibiotic, to upplement their activity intead of replacing it altogether, Berrie (007). Nowaday, many people may think bacteria are aociated with unhealthy ubject only which i not at all. Thi reearch wa carried out to examine bacteria aociated with hand palm and armpit of healthy ubject, and to determine the in-vitro antibiotic enitivity pattern of the bacteria encountered from the hand palm and armpit of the healthy ubject from Adekunle Ajain univerity Akungba-Akoko, Ondo State, Nigeria. Material and Method Study Area The tudy area comprie the town known a Akungba Akoko in Ondo State, Nigeria where a tate government owned univerity i ited. Study Population Thi tudy wa executed among a few undergraduate from the cital del of higher learning in Akungba Akoko. So, the ample were collected from twelve healthy tudent of Adekunle Ajain Univerity Akungba-Akoko, Ondo State, Nigeria. Seven hand palm wab and five armpit wab at Akungba-Akoko. The individual whoe ample were collected age range between 8 and 30 year old. Material Ued The material ued in thi tudy include petri-dihe, wire loop, terile wab tick, buren burner, autoclave, microcope, lide, oil immerion, McCartney bottle, tet tube, yringe and needle a well Copyright IJASR All right reerved Page 5

6 a antibiotic enitive dic which comprie augumentin (Aug), tetracycline (Trt), cloxacillin (Cxc), chloramphenicol (Chl), treptomycin (Str), cotrimoxazole (Cot), erythromycin (Ery) and gentamicin (Gen). Media and Reagent Nutrient agar, tryptone oya agar, eoin methylene, Mueller-Hinton agar, blood agar, mannitol alt agar, crytal violet, afranin, hydrogen peroxide, ethanol and aline water. Sterilization Technique Glaware uch a petri-dihe were wahed with oap and rined thoroughly with clean water and were allowed to air-dry for few minute. They were arranged in the caniter and put in hot air oven for two hour at 70 0 C. The media were terilized uing the autoclave at O C for 5 minute. Collection of Sample The ample were collected uing terile wab tick. Different wab tick were ued to gently rub the palm of the hand and armpit of the healthy ubject. The ample were collected within the hortet pace of time and tranported to the laboratory for further microbiological analyi. Media Preparation The media ued for the iolation of organim were nutrient agar, eoine methylene blue, mannitol alt agar, Mueller-Hinton agar, and all which were prepared following tandard method. Method of iolation With the aid of a terile loop, a mall or loopful portion of pure culture wa taken from incubated tryptone oya agar medium and treaked again on the plate containing mannitol alt agar, eoin methylene blue. Thee plate of media were then incubated aerobically at 37 C for 4hour or more until growth wa oberved. After incubation, the cultural and morphological characteritic of ditinct, well iolated colonie were tudied. Thee included the hape, ize, elevation, edge, opacity, urface and colour repreentative and well eparated colonie obtained from ample were picked, heat fixed and gram tain. Stock culture of pure iolate were labelled accordingly and tored. Method of identification Preliminary characterization of bacteria iolate wa baed on gram tain, morphology and culture. Furthermore, characterization wa carried out with variou biochemical tet. Thee tet included catalae, coagulae, indole production, citrate utilization, tarch hydrolyi and growth characteritic on MSA and EMB agar, and blood agar haemolyi. All media were prepared according to the manufacturer pecification and terilized unle where tated otherwie at C for 5minute. Gram Staining Technique A terile wire loop wa ued to place a drop of terile water on a greae-free lide. The loop wa flamed and allow to cool, it wa ued to pick an inoculum from the plate and placed on the lide. The inoculum wa gentily emuified with the water forming mear. Thi wa then heat fixed by paing it over the flame.the mear werethen flooded with crytal violet and left to tand for 60 econd and were rined with water. Lugol Iodine olution wa poured on the lide and allowed to tay for another 60 econd and later rined with water. The mear were then decoulorized with 95% ethanol which wawahed off immediately. The mear were counter tained with Safranin olution for 30 econd. The lide finally rined with water and then allow ed to air dry. After air drying, the lide were then viewed under the micocope uing oil immerion (X00objective len).organim that retain the purple colour of crytal violet are the Gram-poitive organim while thoe that retain red or Copyright IJASR All right reerved Page 6

7 pick colour of franing are Gram-negative organim. Other morphological characteritic were alo examined and recorded a hort or long rod, cocci in cluter or in chain. Catalae Tet A terile wire loop wa ued to tranfer one drop of hydrogen peroxide on greae-free lide. A mall portion of the inoculum of 4 hour old culture wa picked and placed on the lide with hydrogen peroxide. The lide wa oberved for effervecence (i.e. production of bubble). The reult wa recorded for each iolate a either Catalae-poitive or Catalae-negative depending on the reaction hown. Poitive reult how the converion of hydrogen peroxide to oxygen and water. Coagulae Tet The tet wa performed with 4 hour old culture. A loopful of normal aline wa placed on a clean lide and a mall amount of the culture wa emuified in the normal aline to get a homogenou upenion. A drop of human plama wa added and mixed for 5 econd. A poitive reaction wa oberved by the formation of eaily viible white clump; while organim without formation of white clump, i a negative for the reaction. Haemolyi Tet All the iolate were treaked on the blood agar and they howed different growth characteritic on the agar. Thi tet wa done to how different kind of haemolyi which include alpha (α), beta (β) and gamma (γ) haemolyi. Some of the iolate teted lyed the red blood cell completely (βhaemolyi), ome lyed it partially (α-haemolyi) other iolate did not lye the red blood cell all (γhaemolyi). Growth Characteritic on Mannitol Salt Agar Mannitol alt agar plate were treaked with the appropriate organim and inoculated at 37 C for 4 hour, after which the plate were examined for growthand fermentation of mannitol. Apparent growth of the organim indicated alt tolerance. Fermentation of mannitol wa indicated by a change in thecolour of the medium background of each colony from red to yellow. Carbohydrate tet Phenol red (0.0g), odium chloride (.0g) and fermentable ugar (.0g) were weighed into a conical flak containing 00ml of nutrient broth. The mixture wa wirled to enure that all the component in it diolved. 5ml of the preparation wa dipened into tet tube containing inverted Durham tube. The tube were covered with cotton wool and Aluminum foil and were terilized uing autoclave at temperature of o C for 5 minute. The ugar ued include glucoe, lactoe and dextroe. In addition, all tet tube were inoculated with repective tet organim aeptically and incubated at 37 o C for 3-5day depending on how fat the organim can utilize the ugar. Change in the colour of indicator (phenol red) from red to yellow indicate the utilization of the ugar (that i poitive tet), and if the ga i detected in the Durham tube, it ignifie the organim produce ga. Control tet wa done for each ugar by not inoculating any tet organim in the ugar preparation. Antibiotic Senitivity Tet Senitivity teting were carried out uing comparative method and baed on Gram taining reaction. The bacterial iolate were ubjected to an antimicrobial teting to determine their enitivity pattern. Preparation of Inoculum Culture of iolated organim on nutrient agar lant were ued to prepare the inoculum uing a terile wire loop. Approximately one iolated colony wa tranffered into 0ml of teriled ditilled water Copyright IJASR All right reerved Page 7

8 cointaining in the tet tube and then mixed properly. The liquid now erve a ource of inoculum containing 06CFU/ml of bacterial upention. Inoculating of Plating Medium A terile cotton woolo wab tick wa dipped into the properly mixed inoculum and exce fluid wa removed by rotating the wab againt the inide wall of the urface of the prepared Mueller-Hinton Agar plate which wa contituted according to manufacturer pecification. The pread wa done in four different plane by rotating the plane at 80 C each time. Thi wa toenure that every part of the plate wa inoculated. Placement of Dic A pair of forcep wa flammed until red hot and cooled and it wa then ued to pick the antibiotic dic and it wa a well ued to place the dic on the inoculated plate. The dic were preed down on the agar o a to be in complete contact with agar. The plate were incubated at 37 C for 8 hour. Reult Identification of iolate wa baed on their cultural, morphological and characteritic. A total of ixteen iolate were cultured from all the eleven ample examined for the incidence of bacteria from hand palm and armpit of healthy individual. Out of the 6 iolate cultured 0(6.5%) were gram poitive cocci while 4(5%) were gram poitive rod and (.5%) were gram negative rod. 4(40%) of the 0 gram poitive cocci were mooth, convex, opaque, yellow with entire edge on tryptone oya agar while (0%) wa opaque, circular, convex white porcelain with entire edge and (0%) were opaque, raied, mooth, white porcelain with entire edge wherea 3(30%) were opaque, raied, mooth, white porcelain with entire edge on tryptone oya agar. Moreover, (50%) of the four gram poitive rod were opaque, raied, irregular with entire edge while (5%) wa tranlucent, raied, creamy with lobate edge and (5%) wa opaque, raied, irregular, curled edge and creamy. However, one (50%) of the two gram negative rod were wa tranparent, raied, irregular with entire edge while the other one (50%) wa tranlucent, raied, irregular, creamy with curled edge. In addition, the growth of the iolate encountered in thi tudy wa oberved on both mannitol alt agar (MSA) and eoin methylene blue (EMB) agar. Hence, the gram tained reaction howed that 7(70%) of the 0 Gram-poitive cocci appeared a cocci in cluter while the remaining 3(30%) appeared a cocci in hort chain. Wherea, two (50%) of all the four Gram-poitive rod appeared a long rod align along their axi in parallel in a manner omewhat imilar to fence pot, one (5%) appeared a hort rod and fat and one (5%) appeared a long chain of cell aligned along axi. Furthermore, all the two gram negative rod in thi tudy were gram tained a long rod. Alo, the two iolate cultured howed large, mucoid, brownih growth on eoin methylene blue. The biochemical reaction of the bacterial iolate encountered in thi tudy were alo tudied. Reult of carbohydrate fermentation tet (Table ) how that out of the thirteen iolate teted, 0(76.9%) reduce glucoe to acid without ga production and only (5.38%) iolate reduce glucoe to acid with ga production, while 9(69.3%) and (.3%) of the iolate teted from lactoe and dextroe repectively. However, only three (8.5%) of the 6 iolate were catalae negative while the ret thirteen (8.5%) iolate were catalae poitive. 4(5%) were coagulae poitive while the (75%) iolate Copyright IJASR All right reerved Page 8

9 were coagulae negative. Out of 3 iolate examined for haemolyi in blood agar, 4(30.77%) were alpha () haemolytic, 8(6.54%) were beta (β) haemolytic and (γ) wa gamma haemolytic. Growth and fermentation of mannitol alt agar (MSA) in nine of the iolate were alo tudied. All the nine iolate teted howed tolerance by growing on the agar while 4(44.44%) of the iolate teted fermented mannitol in the agar. Meanwhile, only two (.5%) of all ixteen iolate were able to grow on eoin methylene blue (EMB) agar while the remaining (87.5%) iolate were not able to grow on the agar. Baed on thee characteritic comparable to thoe enumerated in Bergey manual (8th edition) and manual of Clinical Microbiology (5th edition), 7(43.75%) were identified a Staphylococci of which 4(57.4%) iolate were Staphylococcu aureu and 3(4.86%) were identified a Staphylococcu pecie. Other bacteria identified were 3(8.75%) treptococcu pecie, (.5%) Corynebacterium pecie, (6.5%) Clotridium pecie, (6.5%) Bacillu pecie, (.5%) and Klebiella pecie (Table ). Table : Profile of Bacterial iolate in relation to ource of Collection Source Bacterial iolate Number and prevalent rate (n= 6) Hand palm Staphylococcu aureu (.5%) Armpit Staphylococcu aureu (.5%) Armpit Staphylococcu pecie 3 (8.75%) Armpit Corynebacterium pecie (.5%) Hand palm Bacillu pecie (6.5%) Hand palm Clotridium pecie (6.5%) Hand palm Streptococcu pecie (.5%) Armpit Streptococcu pecie (6.5%) Hand palm Klebiella pecie (.5%) Total 6 (00) Table : Incidence of Gram poitive and Gram negative rod from Hand palm and Armpit Bacterial iolate Number and prevalent rate ( n=6) Corynebacterium pecie (33.33%) Bacillu pecie (6.67%) Clotridium pecie (6.67%) Klebiella pecie (33.33%) Total 6 (00) Table 3: Incidence of bacteria from Hand palm and Armpit of Healthy Individual In Relation to Age Age (year) Number of Individual Frequency of bacteria 8- (8.8%) 4 (5%) -5 6 (54.55%) 7 (43.75%) 6-30 (7.7%) 5 (3.5%) Total (00) 6 (00) Copyright IJASR All right reerved Page 9

10 Table 4: Frequency of bacteria cultured from Hand palm and Armpit in relation to Age and Sex Age Sex Frequency of bacteria Male Female 8- (4.9%) (5%) 4 (5%) -5 4(57.4%) (50%) 7 (43.75%) 6-30 (8.57%) (5%) 5 (3.5%) Total 7 (00) 4 (00) 6 (00) Table 5: Ditribution of bacteria in relation to age, ex and ource of collection Age Sex Source of collection Prevalent rate (%) 8- M Hand palm (.5%) 8- F Armpit (6.5%) 8- M Hand palm (6.5%) -5 M Hand palm 4(5%) -5 M Armpit (6.5%) -5 F Hand palm (6.5%) -5 F Armpit (6.5%) 6-30 M Hand palm (.5%) 6-30 F Hand palm (.5%) 6-30 M Armpit (6.5%) Total 6 (00) M Male F- Female Table 6: Antibiotic reitance of the bacterial iolated from hand palm and armpit Iolate code Identification Antibiotic to which iolate were reitance HP a Streptococcu pecie Aug, Str, Tet, Cot, Chl, Cxc, Ery, Gen HP b Klebiella pecie Aug, HP Klebiella pecie Nt HP 3 Clotridium pecie Aug,Tet, Cxc HP 4a Staphylococcu aureu Aug, Cxc, Cot HP 4b Streptococcu pecie Aug, Cxc HP 5 Bacillu pecie Aug HP 6 Staphylococcu aureu Aug, Str, Tet, Cot, Chl, Cxc, Ery, Gen AP a Staphylococcu pecie Aug, Cxc, Chl,Cot, Ery Copyright IJASR All right reerved Page 0

11 AP b Streptococcu pecie Aug, Cot, Ery AP Corynebacterium pecie Aug, Cxc, Cot AP 3a Corynebacterium pecie Aug, Cxc, Cot AP 3b Staphylococcu pecie Nt AP 4a Staphylococcu aureu Aug, Cxc, Cot, Ery AP 4b Staphylococcu pecie Aug AP 5 Staphylococcu aureu Aug, Cxc, Cot, Ery HP Hand Palm AP - Armpit augumentin (Aug), tetracycline (Trt), cloxacillin (Cxc), chloramphenicol (Chl), treptomycin (Str), cotrimoxazole (Cot), erythromycin (Ery), gentamicin (Gen) Nt Not teted Table 7: pattern of reitance of bacterial iolate againt antibiotic employed Bacte rial iolate Numbe r of iolate teted Aug Gen Cxc Chl Cot Tet Str Ery Staph ylococ cu aureu Strept ococc u pecie Staph ylococ cu pecie Coryn ebacte rium pecie Clotri dium pecie Bacill u pecie (00) 3 (00) (00) (00) (00) (00) (5) (33.33) 0(0) 0(0) 0(0) 4 (00) 3 (00) (50) (00 ) (00) (5) 4(00) (5) 3(75) Nt (33.33) (33.33) (33.33) (33.33) (66.6 7) (50) (50) 0(0) 0(0) (50) 0(0) (00) 0(0) 0(0) 0(0) 0(0) 0(0) (00) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) Copyright IJASR All right reerved Page

12 Klebi ella pecie Total 4 (00) 0(0) Nt Nt 0(0) 0(0) Nt Nt Augumentin (Aug), tetracycline (Trt), cloxacillin (Cxc), chloramphenicol (Chl), treptomycin (Str), cotrimoxazole (Cot), erythromycin (Ery), gentamicin (Gen), Nt Not teted Table 8: The profile of multiple antibiotic reitance pattern of iolate Bacte rial iolate Number of iolate teted Staph ylococ cu aureu Strept ococc u pecie Staph ylococ cu pecie Coryn ebacte rium pecie Clotri dium pecie Bacill u pecie Copyright IJASR All right reerved Page

13 Klebi ella pecie Information relating to the ample analyed A total of eleven individual, coniting of 7(63.64%) male and 4(36.36%) female between age 8 and 30 year old were examined during thi tudy (Table 3 and 4). Alo, the ditribution microbe aociated with the ite under conideration wa later on dicovered to be either normal flora uch a Staphylococci or contaminant Klebiella pecie from the environment uch a lower part of urinary tract. Depite the fact that Staphylococci are native or normal flora of the body they alo erve a opportunitic pathogen. Hence, from thi tudy, the incidence of Staphylococcu aureu 4(5%) were found to be the highet, followed by that of coagulae-negative Staphylococcu pecie and Streptococcu peciewhich were (8.75%) each a hown in table. Moreover, there the incidence of Corynebacterium and Klebiella pecie contituted (.5%) each, while Clotridium pecieand Bacillu pecie are accounted (6.5%) each the lowet incident rate (Table ). In addition, the prevalence of bacteria among individual between age and 5 year contituted the highet which wa 43.75% while among individual between age 8 to and 6 to 30 year contituted 5% and 3.5% repectively (Table 3). Therefore, among the ample analyed individual between age 8-, 4(5%) were cultured from a male (50%) and a female (50%). For age range -5, 7(43.75%) iolate were cultured from 6 individual of which 4(66.67%) were male and (33.33%) were female. For age range 6-30 year, 5(3.5%) from 3 individual, (66.67%) male and (33.33%) female (Table 3 and 4). Nonethele, incidence of bacteria i higher in male than female epecially among individual between age -5 year old (Table 4). In contrat, the incidence i lower among individual between age8-. Gram poitive cocci were found be the mot common organim recovered, then gram poitive rod followed by gram negative rod a hown in table 3. Antibiotic reitance pattern of the bacterial iolate A total of fourteen iolate were teted with variou antibiotic enumerated in the material and method. Out of thee fourteen iolate two were reitance to one antibiotic, while five and two iolate were reitance to eight antibiotic repectively. Alo another two were reitance to four antibiotic while only one iolate wa reitance to five of the antibiotic employed (Table 7). The pattern of reitance of each iolate againt the antibiotic teted were evaluated and are hown in table 8. The incidence of Staphylococcu aureu to augmentin, cloxacillin and cotrimoxazole wa highet involving all the four iolate teted. Thi wa followed by reitance to erythromycin occurring in three of the four iolate. Only one Staphylococcu aureu wa reitance to eight of the antibiotic which include augmentin, gentamicin, chloramphenicol, tetracycline, treptomycin, cotrimoxazole, erythromycin and cloxacillin. Furthermore, all the three iolate of Streptococcu pecie teted were reitance to augmentin and cloxacillin while only one of the three iolate wa reitance gentamicin, chloramphenicol, tetracycline, treptomycin and cotrimoxazole. Wherea only two of the three iolate were reitance to erythromycin. With regard to corynebacterium pecie, all the two iolate augmentin, cloxacillin and cotrimoxazole. In contrat, all the two iolate were enitive to gentamicin, chloramphenicol, tetracycline, treptomycin and erythromycin a hown in table 6 and 7. Copyright IJASR All right reerved Page 3

14 Meanwhile, the Clotridium pecieencountered in thi tudy wa reitance to augmentin, cloxacillin and tetracycline while the Bacillu pecie encountered how reitance to only augmentin and enitive to gentamicin, chloramphenicol, tetracycline, treptomycin, cotrimoxazole, erythromycin and cloxacillin. With regard to Klebiella pecie, all the iolate teted were reitance to augmentin and amoxicillin. Hence, the two iolate of Klebiella pecie were enitive to gentamicin, ofloxacin, nalidixic acid, cotrimoxazole, nitrofuratoin and tetracycline (Table 7 and 8). The profile of multiple antibiotic reitance pattern The profile of multiple antibiotic reitance of the iolate teted wa tudied. Out of the fourteen iolate evaluated two iolate were reitance to one antibiotic, one Staphylococcu pecie and one iolate of Bacillu pecie. Alo, two iolate were reitance to two antibiotic the iolate were Streptococcu pecie and Klebiella pecie. In addition, one iolate of Staphylococcu aureu and Streptococcu pecie howed multi-reitance to eight of the antibiotic employed. Five iolate of which two were Corynebacterium pecie and one Staphylococcu aureu, Streptococcu pecieand Clotridium pecie howed multi-reitance to three of the antibiotic employed. However, the only iolate that howed multi-reitance to four of the antibiotic were two of the four Staphylococcu aureu. While of one iolate of the Streptococcu pecie howed multi-reitant to five of the antibiotic. Dicuion The microbiology of hand palm and armpit i an important apect of the area that need pecial attention in medical microbiology becaue of the function of hand palm and armpit in preading of infectiou agent and becaue of the odour produce under the arm a a reult of the bacteria that the two ite of the body in under conideration. In human, bacteria are the mot common and numerou component of the kin flora; even though, human kin i a remarkable organ, the larget but often taken for granted, Neter et al. (007). At hand palm and armpit, a total of 6 iolate were recovered from individual a hown in table 3. The reult from thi tudy howed that ome dieae and pathogen i highly pecific and may be due to any everal caual agent at the ite of the body. Therefore, dieae doe not neceary follow expoure to a given caual agent. In fact, factor including the degree of reitance of the hot and virulence of the pathogen. Thi finding i in agreement with the report by Singleton Paul and Neter and other (Singleton, 997; Neter et al., 007). However, microorganim which colonize a given habitat affect each other in variou way becaue they will have to compete with each other for nutrient, oxygen, and pace. According to the report by Singleton, thoe that cannot urvive will have to be eliminated from uch habitat, a it can be een from thi tudy where mot of the iolate 0 (6.5%) were gram poitive cocci, Singleton, (997). Further invetigation revealed that weat from the armpit remain terile when ecreted but the activitie of the native flora of the part of the body break the apocrine and thereby reponible for the unpleaant odour. A part of the bacteria encountered in thi tudy, Corynebacterium pecie (.5%) and other native flora uch a Staphylococcu aureu (5%) that are conitently found on a pecific area of the body uch armpit have made ite of the body ource of body odour a a reult of the releae of chemical called 3-methyl--hexenoid acid by the native microbe, Alkerman et al. (005). Moreover, according to finding in thi tudy, not many are aware of the caue of armpit odour of which the main caue i the culprit bacteria which reide in the weat on the kin. Thi finding i alo in agreement with the report by Alkerman and other which revealed that ome individual ha odour all over their body, Alkerman et al. (005). However, infection can be directed toward the Grampoitive cocci ten (6.5%) and mot common two (.5%) Gram-poitive rod iolate encountered from hand palm and armpit repectively. Copyright IJASR All right reerved Page 4

15 In addition, incidence of Staphylococcu aureu, Streptococcu pecieand Staphylococcu pecie were the predominant organim iolated from the two ite of the body contituting (5%), (8.75%) and (8.75%) repectively (Table ). Thi obervation confirm the finding by Precott et al (008) that Staphylococci are pathogen that inhabit the membrane of kin and feeding on the weat and dead kin. However, chance of infection by hand palm i high a a reult of the gram poitive rod iolate from the ite of the body, ince Bacillu pecie and Clotridium pecie contituting (6.5%) each. Thi mut have being due to infection by air, water or oil contamination by the two iolate. Alo, Klebiella pecie(.5%) iolated from hand palm mut have a well be the reult of contamination from the environment; becaue Klebiella pecie i one of the native flora of the lower part of the intetinal tract. The reult of the age ditribution of the individual examined in thi tudy how that 54.55% of the participant were around age -5 year with 7(43.75%) bacterial incidence from 4(66.67%) male and (33.33%) female (Table 5). Thi invetigation reveal that both men and women have thee bacteria on their armpit and produce mulder odour thi i motly common among young adult. The reult obtained how the incidence of bacterial iolate to be higher in male (66.64%) than female (36.36%). The reult i not apparently clear but could be aociated with the nature of the ampling populated examined. The antibiotic enitivity teting on bacterial iolate cultured from the body ite viz: hand palm and armpit among the individual between age eighteen and thirty year old wa carried out to provide uggetion for practicing clinician on the antibiotic to which thee organim were enitive. Although, reitance of infectiou microorganim of hand and armpit againt antibiotic are regularly reported, it i difficult to compare reult ince variation in methodology may contribute to ome extent to thee difference. Therefore, it follow that the clinician who manage infection of the hand palm and armpit hould be acutely aware of the changing antibiotic enitivity pattern of the bacteria that caue uch infection. In addition, the in-vitro antibiotic enitivity teting revealed that the Staphylococcu aureu train had coniderable reitance to many of the antibiotic employed. Reitance to augmentin (00%), cloxacillin (00%) and cotrimoxazole (00%) were particularly triking. With regard to Klebiella pecie which happened to be the only Gram-negative bacteria from thi tudy,all the two iolate encountered how reitance to augmentin (00%) a well a amoxicillin, hence thi imilar finding in which all Klebiella pecie were reitant to amoxicillin ha been reported by Monica (004). With regard to Bacillu pecieonly one iolate cultured wa reitance to only augmentin. In cae of Corynebacterium pecie, the in-vitro antibiotic enitivity teting howed that all the two iolate from thi tudy were (00%) multi-reitance to augmentin, cloxacillin and cotrimoxazole. Meanwhile, the reult obtained how that only one the Staphylococcu aureu and Streptococcu pecie teted howed reitance to eight of the antibiotic employed. Wherea, Clotridium peciefrom how 00% multi-reitance to augmentin and tetracycline. Thi finding i alo in agreement with the report by Monica Chebrough that miue of drug againt infectiou organim can lead to multi-reitant of mot the antibiotic employed againt the infectiou agent, Monica (004). In general, from thi tudy, it i apparent that many of the organim iolated were reitant to many of the antibiotic employed with a coniderable number being reitant to a many a three, four and even five antibiotic. The high incidence of multiple antibiotic reitance in thee organim indicate an alarming cae of multi-reitance and i of concern, becaue the information obtained Copyright IJASR All right reerved Page 5

16 from the report by Monica that mot of the antimicrobial reitance which i now making it difficult to treat ome infectiou agent i due to the extenive ue and miue of drug. Therefore, the abue of antibiotic could not be implicated a being reponible for the high incidence of antibiotic reitance encountered in thi tudy. Concluion Thi tudy how that bacterial iolate particularly, gram poitive cocci and gram poitive rod were predominant on the kin. A a matter of fact, Staphylococcu aureu were predominant pathogen aociated with hand palm and armpit part of the body while Streptococcu pecie and Staphylococcu pecie were found to be predominant at the ame rate. Thi make it eaier for Staphylococcu to invade wound epi whenever there i cut in the body becaue Staphylococcu i normal or native flora of the body but become pathogenic under certain condition a opportunitic infection uch a in the cae it infection on wound epi. Infection of hand palm and armpit i preventable by taken into conideration the genera uch a Clotridium which may bring about infection from food a it can be een in the cae of Clotridium perfrigen or oil a in the cae Clotridium tetani. Thu, the infection from thee two genera i preventable becaue of the aetiologic factor uch a poor hand and armpit hygiene which caue food infection and body odour repectively. Therefore, education and awarene of people on keeping proper hygienic condition through proper hand wahing technique with oup or detergent and daily wahing body with oup will therefore ameliorate bacterial infection by hand and body odour from the armpit. Antibiotic uceptibility pattern of the bacterial iolate from the tudy indicate the preence of highly reitant pecie of bacteria which ugget the reliance on pecific uceptibility pattern of bacteria for treatment. Some of the antibiotic uch a gentamicin, chloramphenicol and tetracycline can till be precribed with high ucce rate, but it i important that reearch in the field on antibiotic uceptibility hould be continued becaue microorganim may vary in different ecoytem. Contribution to knowledge Thi tudy reveal that bacteria were prevalent in the two ite of the body under conideration. The invetigation alo unveiled a high incidence of bacteria among people between the age of twenty-two and twenty-five year, thi could be due to the level of their hygiene. Beide, the reult of the invitro uceptibility teting on commonly employed antibiotic uch a gentamicin, treptomycin, tetracycline and erythromycin may not be effective for in treatment of hand and armpit infection. Therefore, information on reitance pattern of iolate encountered in the tudy to commonly available antibiotic employed will ait the clinician in making improvement in management of hand and armpit hygiene. REFERENCES. Alkerman, A. B., Almut, B., Bruce and Geoffrey, J. G. (005). Hitology Diagnoi of a. Inflammation Skin Algorithm Method Baed Pattern Analyi. ISBN Akiyama, K., Hiramatu, N., Igari, J., Koyama, T. and Oguri, T. (00).Jpn J Antia. biot.55(): Baron, S. (996). Medical Microbiology. 4th edition. Galveton (TX): Univerity of Texa a. Medical Branch at Galveton. NBK8035PMID: Beck-Sague, C., Giuliano, D. and Villarino, E. (994). Infectiou dieae and death among a. nuring home reident: reult of urveillance in 3 nuring home. Infect Control Hop Epidemiol. 5:494. Copyright IJASR All right reerved Page 6

17 5. Bergeron, M.G. (995). Treatment of pyelonephriti in adult. Med Clin North Amer. 79: Berrie, C. (007). Carbapenem-Reitant Klebiella pneumoniae Outbreak in an Iraeli a. Hopital. Medical New. 7. Bingen, E. (994). Application of molecular method to epidemiologic Invetigation of a. noocomial infection in a pediatric hopital. Infect Control HopEpidemiol Brun-Buion C. and Legrand, P. (994). Cantopical and non aborbable antimicrobial a. prevent cro tranmiion of reitant train in ICU? Infect Control HopEpdemiol.5: Cogen, A.L., Nizet, V., Gallo, R.L. (008). Skin microbiota: a ource of dieae or defence? a. Br. J. Dermatol.58(3): Cohan, F. M., Coenye, T., Keil, E. J., and Stanckenbrandt, E. (005). Re-evaluating a. proakaryote pecie. Nature Rev. Microbiol3: Duncan, W. C., Knox, J. M. and McBride, M. E. (975). Phyiological and environmental a. control of Gram negative bacteria on kin. Brit. J. Dermatol. 93: Foxman, B., Palin, K.and Zhang, L. (995). Bacterial virulence characteritic of a. Echerichia coli iolate from firt-time urinary tract infection. Infect Di. 7: Freter, R., Botney, M. and Brickner, J. (983). Survival and implantation of Echerichiacoli a. in the intetinal tract.infect Immun. 39: Granato, P. A.(003). Pathogenic and Indigenou Microorganim of human. In. Manual a. of Clinical Microbiology, 8th ed., ASM Pre. 5. Johnon, J. R. and Stamm, W.E. (989). Urinary tract infection in women: diagnoi and a. treatment. Ann Intern Med. : Kathleen,P. T. (005). Foundation in Microbiology. 5th ed. McGraw-Hill, Inc., a. Avenue of the America, New York, NY Kenneth Todar. (008). A textbook of Bacteriology. Madion-Wiconin TheGood, the Bad, a. and The Deadly Scince Magazinevol. 304; Pg Kuciec-Tepe, N., Bejuk, D. and Kouta, D. (006).Acta Med Croatica. 60(4): Lledo, W., Hernandez, M. and Lopez, E. (009). Guidance for Control of Infection with a. Carbapenem-Reitant or Carbapenemae-Producing Enterobacteriaceae in Acute Care Facilitie.58(0); Lockhart, S.R., Abramon, M.A., Beekmann, S.E., Gallagher, G., Riedel, S., Diekema, D.J., a. Quinn, J.P. and Doern, G.V. (007). J.ClinMicrobiol.45(0): Monica, C. (004). Antimicrobial enitivity teting. Ditinct Laboratory Practice in Tropical a. Countrie part (Cambridge low price edition). ISBN Murray, B. E. (990). The life and time of Enterococcu. Clinical Microbiology. Rev. a. 3: Nina, C. (009). &A Literature Review of the Practical Application of Bacteriophage a. Reearch&, 84p. 4. Peter, M. H. and Annie, M. J. (009). The changing epidemiology of reitance J. Antimicrob a. chemother. 64. Vo. 4. Pp Podchun, R. and Ullman, U. (998). &Klebiella pp. a Noocomial Pathogen: Epidea. miology, Taxonomy, Typing Method, and Pathogenicity Factor&. Clinical Microbiology Review (4): Precott, L. M., Harley, J. P. and Klein, D. A. (008). Genetic and molecular biology. a. General Microbiology (7th edition). McGraw-Hill, New York; pp Precott, L. M., Harley, J. P. and Klein, D. A. (008). Normal microbiota of the human body: a. General Microbiology, 7th edition. McGraw-Hill Companie Inc. New York; pp Rotter, M. L. and Koller, W. (990). Surgical hand diinfection: Effect of equential ue of Copyright IJASR All right reerved Page 7