Advances in Bioscience and Bioengineering ISSN 2201-8336 Volume 1, Number 2, 2013, 44-53 The Effect of Hand Treatments on Staphylococcus Aureus: A Normal Flora of the Human Palms Ajayi, Adesola Adetutu; Aiyedun, Bankole Titilope; Olasehinde, Grace Iyabo Department of Biological Sciences, Covenant University, Ota, Ogun State, Nigeria Corresponding Author: Ajayi, Adesola Adetutu, Department of Biological Sciences, Covenant University, Ota, Ogun State, Nigeria Abstract. A total of forty samples of hand swabs obtained from female and male students palms at Covenant University, Ota were studied and examined for the presence of S. aureus. The samples were inoculated on Mannitol Salt agar (MSA) and incubated at 37 o C for 24h and 48h. Colonies which appeared yellow on MSA were subjected for further tests. Macroscopic examination indicated that the organism was Gram positive after the Grams staining technique and that they appeared in clusters and spherical (coccoid) in shape under the microscope. The isolates were characterized using several biochemical tests. The organism fermented Glucose, Lactose, and Sucrose with acid production without gas and Starch with gas production only. The isolates were susceptible to Ciprofloxacin, Gentamycin, Pefloxacin and Amoxycillin but resistant to Erythromycin, Ceftriazone and Cotrimoxazole antibiotics. The effect of hand treatments on the microbial flora of the palms were studied and washing of ha nds thoroughly with soap, water and using an alcohol based hand sanitizer can prevent the spread of staphylococcus skin infections. Keywords: Staphylococcal Infections, Normal flora of the skin, Staphylococcus aureus, Hand treatments Introduction Staphylococcus aureus is one of the most common causes of nosocomial and community-acquired infection. The normal flora of individuals is colonized by microorganisms especially bacteria (Falang et al., 2002). The skin is inhabited by a large number of microorganisms because of its exposure to the environment (Faden et al., 2007; Saxena et al., 2010). S.aureus had been identified as one of the common bacteria inhabiting the skin (Baker, 2010). The bacteria found on the skin have important interactions with host cells and they therefore serve Copyright 2013 the authors. 44
45 Advances in Bioscience and Bioengineering various functions (Ruimy et al., 2010). Staphylococci produces various enzymes such as protease, lipase and hyaluronidase which aids the infection process (David and Daum, 2010; Pickett et al., 2009). Staphylococcus aureus is one of the most common causes of nosocomial and community-acquired infection (Lyytikainen et al., 2005). Humans are a natural reservoir of S.aureus. Von Eiff et al. (2001) reported that S. aureus is found on the face and hands particularly in individuals who are nasal carriers. Gordon (1998) reported S. aureus as the causative agent for a variety of diseases and associated its success as a pathogen to its ability to cause such a wide range of infections to the presence of extensive virulence factors. The infection associated with S.aureus is largely due to its presence as a normal resident of the skin. They live harmlessly on many skin surfaces especially around the nose, mouth, genitals and anus but they cause an infection when any part of the skin is punctured or broken for any reason (Van Belkum et al., 2009).Although S. aureus is a normal inhabitant of the skin, mucous membranes, respiratory and gastrointestinal tracts, it can invade any organ or system to produce infection ranging from localized to invasive diseases (Bartlett and Hulten, 2010). Some of the localized diseases include furuncle, boils and impetigo which are community acquired while invasive infections include septicaemia, osteomyelitis, arthritis, pneumonia and endocarditis which are hospital acquired. This study therefore was carried out to isolate and characterize S. aureus associated with the palms of male and female students of Covenant University, Ota with a view of studying the effect of hand wash and sanitizers on S. aureus in relation to the sex or gender of the students. The antibiotic susceptibility test was carried out on the species of S. aureus isolated. Materials and Methods Collection of samples The samples employed for this research work were taken from twenty male and female students of Covenant University, Ota, Ogun State. Samples were collected from each student s palms using sterile swab sticks. The swabs containing the inoculums were then streaked on sterile Mannitol Salt agar and Nutrient agar in sterile petri dishes for the growth and isolation of different
Advances in Bioscience and Bioengineering 46 distinct colonies. The inoculated plates were then incubated at 37 o C for 24h. After incubation, the colonies appearing yellowish or with were isolated and subcultured onto nutrient agar slants for further characterization. Biochemical Tests The biochemical tests carried out were Grams test, catalase test, coagulase test, citrate utilization test, urease test, indole test, methyl red test, Voges proskauer test, starch hydrolysis and sugar fermentation test according to the methods described by Adejuwon et al. (2012). Antibiotic susceptibility test The bacterial sensitivity was carried out on pure cultures of the organisms. The disc diffusion method was used to determine the susceptibility and resistance of the organisms to the antimicrobial drugs. The inoculums from the nutrient agar slant was removed by means of sterile wire-loop streaked on the Mueller Hinton agar plates ensuring that the inoculum was spread over the surface of the medium using a sterile spreader. A Gram positive antibiotic multo disk were aseptically removed and placed at the centre of the plate. The forceps were used to press the discs firmly to the medium. The plate was incubated at 37 o C for 24h in an inverted position. The zones of inhibition formed around each antibiotic disc were measured after the incubation period. Use of Questionnaire A total of forty samples of hand swabs obtained from female and male students palms were studied for the presence of S. aureus. The isolates were characterized using several biochemical tests. The bacterial sensitivity tests were carried out on pure cultures of the organisms. The disc diffusion method was used to determine the susceptibility and resistance of the organisms to antibiotics. Results The swabs from students palms streaked on Mannitol Salt agar revealed colonies which fermented mannitol and appeared golden yellow. The cultural and
47 Advances in Bioscience and Bioengineering morphological characteristics (Table. 1) showed that the colonies were spherical in shape; they appeared in clusters and are non-spore forming, Gram positive organisms. The species of S. aureus isolated were characterized using Catalase test, Coagulase test, Citrate utilization test, Urease test, Indole production Test, Starch hydrolysis and Methyl red, Voges Proskauer test (Table. 2). The organism fermented Glucose, Lactose, and Sucrose with acid production but without gas. There was neither acid nor gas production with Starch (Table. 3). The isolates were susceptible to Ciprofloxacin, Gentamycin, Pefloxacin and Amoxycillin but resistant to Erythromycin, Ceftriazone and Cotrimoxazole.The sex distribution of the forty students examined revealed the differences in students affected by staphylococcus infections (Table. 5), staphylococcal illnesses (Table. 6) and the sex distribution based on the different hand treatments employed by students in hand washing at Covenant University, Ota (Table. 7). Table 1: Morphology and Cultural Characteristics of Staphylococcus aureus associated with students palm at Covenant University, Ota Features Colour Elevation Margin Surface Light Transmission Grams reaction Appearance Yellow Convex Entire Smooth and Shiny Opaque + (Purple colour), Spherical (Coccoid), in pairs, Short Chains or Grape-like Clusters
Advances in Bioscience and Bioengineering 48 Table 2: Biochemical Characteristics of Staphylococcus aureus associated with Students palms at Covenant University, Ota Biochemical Tests Specie 1 Specie 2 Citrate Utilization Test + + Coagulase Test + + Indole Production - - UreaseProduction + + Methyl Red Test + + Voges proskauer Test - - Starch Hydrolysis Test + + Catalase Test + + Keys + positive - negative Table 3: Sugar Fermentation Test on Staphylococcus aureus associated with Students palms at Covenant University, Ota Sugars Glucose Galactose Lactose Maltose Sucrose Starch Keys A Acid production Reaction A A A A A N/A N/A No Acid production
49 Advances in Bioscience and Bioengineering Table 4: Antibiotic resistance profile of the species of Staphylococcus aureus with zones of Inhibition Antibiotics Quantity Zone Diameter Zone Diameter (mm) Code (UG) (mm) Species 1 Species 2 AMX 25 14 (CI) 15 (CI) OFL 5 9.0 7.0 STR 10 3.0 3.0 CHL 30 4.0 5.5 CEF 30 2.5 3.0 GEN 10 21 (CI) 20(CI) PEF 5 16 (CI) 14 (CI) COT 25 2.0 3.0 CPX 10 20 (CI) 24 (CI) ERY 5 2.5 2.0 Keys AMX Amoxycillin OFL Ofloxacin STR Streptomycin CHL Chloramphenicol CEF Ceftriazone PEF - Pefloxacin COT Cotrimoxazole ERY - Erythromycin GEN -Gentamycin CPX Ciprofloxacin CI Complete Inhibition
Advances in Bioscience and Bioengineering 50 Table 5: Infections Sex Distribution of Students affected by Staphylococcal Infections Number of Frequency (%) Number of Frequency (%) Male Students Female Students Boils 11 55 5 25 Pimples 3 15 13 65 Stye 6 30 2 10 Total 20 100 20 100 Table 6: Sex Distribution of Students with Staphylococcus Illness Illnesses Number of Frequency (%) Number of Frequency (%) Male Students Female Students Common Cold 8 40 10 50 Flu 8 40 4 20 Diarrhea 4 20 6 30
51 Advances in Bioscience and Bioengineering Table 7: Sex Distribution of hand treatments employed by students for hand washing at Covenant University, Ota Hand Number of Frequency (%) Number of Frequency (%) Treatments Male Students Female Students Water only 8 40 2 10 Water and 4 20 4 20 Soap Water and 3 15 6 30 Antiseptic (Dettol) Water and 2 10 5 25 Hand Sanitizer Liquid Hand 3 15 3 15 wash and Water Discussion The results of this investigation revealed that the two species of Staphylococci investigated were coagulase positive and coagulase negative. The ability of S. aureus to produce coagulase have been taken as customarily an in-vitro reaction indicating its potential virulence since coagulase production is characteristic of the majority of Staphylococci that cause diseases (Bartlett and Hulten, 2010). The questionnaires administered to male and female students of Covenant University revealed that the percentage of students who use water as a cleaning agent usually have diarrhea. The survey also revealed that majority of the respondents was familiar with the infections caused by Staphylococci but lack adequate knowledge of the disease transmission and appropriate hand-washing techniques. This can be attributed to students knowledge, attitude and perception of the presence of S. aureus as part of the normal flora of the skin.
Advances in Bioscience and Bioengineering 52 Majority of the students are also not familiar with the causative agents of the diseases but attributed them to stress, malnutrition, skin types, adolescent period, diet, menstrual cycle, change of soap, cosmetics, cleansers and climatic conditions. Conclusion The importance of basic hand washing techniques cannot be overemphasized because of its effectiveness in preventing the transmission of Staphylococcus aureus. Washing of hands thoroughly with soap and water, using an alcohol based hand sanitizer, keeping cuts and scrapes clean, covering them with bandages until it is healed, avoiding contact with wounds, avoiding the sharing of personal items such as towels, razors, clothing or equipment. Cleaning surfaces where bare skin rubs such as exercise equipment, wrestling mats and surfaces in the home and showering with soap and water after working out or playing sports prevents the spread of Staphylococcus infections. REFERENCES [1] Falang, V., Grinnell., F., Gillchrest, B., Maddox, Y.T. and Moshell,A. (2002). Journal of Applied Bacteriology 21: 125-127 [2] Faden, H., Rose, R., Lesse, A., Hollands, C., Dryja, D. And Glick, P.L. (2007). Clinical and molecular characteristics of staphylococcal skin abcesses in children. Journal Pediatric 151 (6): 700-703 [3] Saxena, S., Thompson, P., Birger, R., Bottle, A., Spyridis, N. and Wong, I. (2010). Increasing skin infections and Staphylococcus aureus complications in children, England 1997-2006. Emergence Infectious Diseases 16 (3): 530-533 [4] Baker, C.J. (2010). Why has methicillin-resistant Staphylococcus aureus become such a successful pathogen? A pediatric perspective. Infectious Diseases Clinic Practical 18 (4): 231-234 [5] Ruimy, R., Angebault, C. and Djossou, F. (2010). Are host genetics the predominant determinant of persistent nasal Staphylococcus aureus carriage in humans? Journal Infectious Diseases 202 (6): 924 934
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