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ISOLATION, IDENT1FICATION AND SENSITIVITY T 83THW OP 3E3S FLORA FR(M KENYATTA B03PITAL 3LAPP \NB . K! BY , ,.ui ftj.B-iIL/i ( h i 3) A DISSERTATION SUBMITTED IN PARTIAL FULFILMENT FOR THE AWARD OF THE BACHALOE OF PHARMACY DEGREE DEPARTMENT 0? PHARMACY FACULTY OF MEDICINE UNIVERSITY OF NAIROBI NAIROBI, KENYA. T3 JUNA4L 19 80 university of NAIROBI Library ■III. 0392015 4 /jy (ii) ISOLATION f IDENTIFICATION AND SENSITIVITY TH .Jf4» OF 3KIN FLORA FROM KENT ATT A HOSPITAL STAFF AND STUDENTS BY UNDER THE SUPERVISION OF MR. A.L. PALEKAR FACULTY OF MEDICINE DEPARTMENT OF PHARMACY UNIVERSITY OF NAIROBI (iii) ti ri.JP A g- A .A.if This piece of work is affectionately dedicated to: My Parents Mr. & Mrs. Salim. My Twin 3i3ter Jamnia and also to my very special friends rialixaa* Salim and Amin. Praise be to Allah for giving me strength and determination to start and finish this project in time. I wish to say special thanks to: Who supervised me during Hr. A.L. Palekar the entire period this project # • »•****# ft * * * Sr & *.' * A * 4 «• « * * * was carried out. I am particularly grateful to him for reading through the 3cript and quidlng me in any alterations and corrections that had to be made. Technical Staff of the - My appreciation particularly to Department of Phnroacy th038 ln tha Aerobiology/ phamaceutics section for then continous assistance without 'whose help the experimental 'work would not have be on finished in time. Special thanks for her tireless Miss Mwarl assistance and kindness. Department of Pharmacy - From whom I collected the akin staff and students microorganism. Mias hunice Iiuthoni - For her fantastic, flawless typing. (v) JLJm-JS-J... JL-Z -3 BAGS SUMMARY.... ..................................1 INTRODUO?10N.................................. 2 EXPERIMENTAL................. ........... 16 R E S U L T S .......................................21 DISCUSSION.................................... 29 CONCLUSION....................................50 REFERENCES 31 1 3 U H M A S .T The purpose of this research project was to isolate ;Acroorganisras from skin surface and identify them using various microbiological techniques. There after their sensitivity was checked against commonly used chemotherapeutic agents. The skin flora specimens were collected from Kenyatta National hospital (Department of Pharmacy), staff and students. As such results obtained refer to this locality. Hence variations may occur if the research was carried elsewhere. Cultures from specific parts of the body (Pore-head, arm, and leg) were done on agar plates. Different colonies obtained were identified and their sensitivity checked. Prom the results obtained, it was found that:(i) The microorganisms were gram-positive (ii) A majority of these were coagulase positive (iii) Some were found to be catalase positive while others were catalase negative. (iv) The microorganisms were found to be most sensitive to Gentamycin and least sensitive to 3ulphafurazole. 2 2 i li 1 a o o v a t 1 o s ..Bacteria have the following general structure. ' srnowM i t %:9 b© j.mta©#p®ia in salso aa4 Io '.. p- ••;* n>raxnhaeh described two- speciesi afxgn -•4 jM t «*tj& alv,:% fuIianeXXe in. 1930 introduced the alau-siideation o f staph .based on differences. in : ^■ ' •? . ,. .; 7... V _ i, ... .. . •. composed of (i) Proteins (ii) Lipids (ill) Carbohydrates and (xv) nucleic acids. ■ H-.j , la.^eriftL.g.eiA Chemi.stiy , ^TM::}0$Xt XV B QV-r< Under all conditions of growth, protein is a raj or component of the bacterial cell. However, the existance of certain classes of conjugated proteins such as phosphoproteins and glycoproteins have not been established in bacteria. Lipoprotein exists in cell Membrane of the bacteria {Hcquillen I960), lipids have been extracted from bacteria. Simple In the main, these are fatty acids, although a few neutral compounds Lave been obtained as well. Carbohydrates occur as intermidiates in biochemical reactions, as intracellular storage material, as a portion of nucleic acid, as part of the cell wall and as extracellular capsules. DUA of bacteria is similar in composition to that of higher animals and plants. Bacterial RHA contains the eonvectional components of ribose, adenine quanin, cytosine and uracil together with minor amounts of unusual bases. 4 However in terms of human disease, the most important genetic changes are those leading to drug resistance. •*#*»9**>**N!»~***m‘ a-.JI s-'.vwv? s --^ «MK*)Rac 10 PBYJIOAI, “ias *oih;t; * i c .;l ,v . Staph are among the most resistant of the non-sparing, bacteria and cati survive many adverse n m r n rental conditions enroute from host to host they highly' resis tant to light, etreiaes of temperature and drying. They are roarkedly resistant to phenols and many other Dcidin te leucocytes 'WQ disinfectants. Cellular Antigent3: »>peeies-specific polysaccharide A and 3 are two cell wall antigens which determine the immunological specificities of staph, aureus and staph. Alfeus respectively. A - polysaccharide from staph aureus has been shown to be composed of reactive residues of N-acetyl glucosamine i.e. Teichoic acid. B - polysaccharide on the other hand was found to be a polyglycerol-phospho Teichoic acid. Product Coagulase Activity Other properties dnsyrne that clots blood plasma Staphylokinase Snsyme that degrades fibrin clot Haemolysin3 All are haemolytic (Toxins) Harrow haemolytic Disrupts mammalian (Alphan) spectrum cells, Is Jertnonec to ti zing, - • lethal Product (Beta) B Activity Other proptertiea Phospholipase, Narrow haenolytic spectrum (Qamiaa) & Broad iiaemolytic Uytotoxic to (delta) spectrum mammalia cells Disrupts mammalian cells Leukocidin Kills leucocytes Two -(interacting proteins and heat labile Eaterotoxin Hyalvaronidase Exfoliation of produced by phage Infant skin group II organisms Enzyme that degrades hyaluronic acid Lipase Enzyme that degrades lipids Proteinase Enzyme that degrades proteins Penicillinase Enzyme that splits the 3-lactam ring . PATHOGiafSCITY Mo3t normal human beings carry large numbers of staphylococci and related organisms both in the nose and on the skin. She relatively non-pathogenic opportunistic staph, epidermidis is almost always found among the normal flora of the skin and mucous membranes of the G.I.X. and respiratory tract. Whereas the pathogenic staph, aureus is a transient, temporary member of the microb'al flora. Sven though staph aureus is often carried by healthy individuals, under certain circumstances it causes severe infection and can kill its host. Because of its frequent presence on the body 3urface3, it is in a position to invade whenever defenses are slightly impaired. Consequently it is the mo3t common cause of both traumatic, and surgical wound infection and superficial 3kin infections. Hature of lesiona: ?he hallmark of staph, diseases is aupp: ioation. Once virulent staph, gain a foothold in deeper tissues if the body, their multiplication causes necrosis and eventual abscess formation. Ituch of the localised tissue damage that results ia irreversible and therefore leads to permanent scarring. Only in unusual severe infections do the organisms break through the localising barrier of the lesions and invade the lymphatics and blood stream. Mechanism of pathogenisitv: Many of the toxins and other substances produced by staph, its pathogenisity. ureusprobably contribute to The precise contributions of these are not known but vary from one infection to another. The outer layer of the cell wall of staph, aureus contains a substance called protein A. demonstrated to be antiphagocytic. This has been It has the unique ability to interact with, the Fc region of lg G, thereby interfering with opsarization involving cytophilic antibodies. Specific-phage receptor-sites are present on the surface of staph, aureus cells. Of the Fom main groups identifiable - by phage typing, strains of group I and III are most often responsible for hospital infection with antibiotic resistant staph. Most strains produce a leukocidin, which apparently acts on the agtoplasmic membrane of many cell types including human neutrophiles and macrophages causing lysis of the cells. X-taxins of virulent staph, is highly toxic for human macrophages (but not neutrophiles), epithelial cells. causes necrosis of skin. It Fnterotoxi.n, is the major factor in the pathogenesis of staph food poisoning. CraOTHXRAPY Chloraraphenical Chemistry Chloramphenicol is a crystalline, neutral and stable compound with the following structure: - % It is highly soluble in alcohol and poorly soluble in water. However, its succinate ester i3 highly soluble in water and ia hydrolysed in tissues with the liberation of free chloramphenicol. Antimicrobial action Chloramphenicol i3 a potent inhibitor of protein synthesis and has little effect on other metabolic functions. It acts on the 50 > units of ribosomes and interferes markedly with the i corporation of amind acids into newly formed peptide. Chloramphenicol is bacteriastatic for many bacteria. action is reversible with removal of the drug. Its Host gram positive bacteria are inhibited by chloramphenicol in concentration 1 - 1 0 % / ml and gram negative bacteria in cone. 0.2-5mg/ial. Resistance In most bacterial species, large population of chloramphenicol susceptible cells contain occasional resistant mutants. These mutants are usually only 2-4 times more resistant than the parent populations. Consequently they emerge slowly in treating the individual. The precise mechanism of chloramphenicol resistance in bacteria mutants has not been established. However, it may be associated with the presence of bacteria ensymes which acetylate the drug clinical uses* Chloramphenicol is a possible drug of choice in ly a few types of infections (1) Symptomatic salmonella infection 9 (2) Haemophilus influenza Meningitis (3) Occasional bacterimia caused by gram negative bacteria Adverse Reaction - (1) Gastroenteritis (2) Bone-marrow disturbanc e3 (3) Toxicity in new b o m s Chemistry: Free tetrcyclinea are crystalline, amphotecic substances of low solubility. They aro available as their hydrochloride which are more soluble giving acidic solutions. Tetrcyclines combine firmly with divalent metal roins, and this chelation can interfer with its absorption and therefore its activity. Antibiotic activity: Tetracyclines are bacteriastatic for many gran and gran negative bacteria. ositive They are effective inhibitors of phosphorylation of protein synthesis; they appear to inhibit the binding of Aminocrcyl -t KNA to 30s units of ribosomes. The basis of their selective action on different organisms is not understood. Resistance: In vitro resistance to tetracycline develop slowly in a atep-wive manner. Bacteria resistance to one tetracyoline usually show resistance to others in the group. retraoycline reelstance organisms are likely to be acquired frora other individual in a population where tetracycline has been widely used. Clinical use. Tetracyclines are the moot typical "Iroud Spectrum’’ antibiotics. Tfcey are effective against a v riety of microorganisms and for this reason, are often U 3 e d indiscriminately. They may be the drug of choice in cholera mycoplastica pheumonia. They are Useful in mixed bacterial infection related to the respiratory tract especially sinusitis and bronchitis, have been used in skin infection particularly ache. verse reactions: (a) G.I.T. disturbances (b) Liver, kidney local tissue toxicity (o) ihotosensitization ■ Chemistry; Streptomycin is a triacidic base consisting of streptidine, streptose and N-methy1-1-glucosamine. / Seve.al salt3 have been prepared from streptomycin, the sulphate being the most widely used. It i3 a white substance with a bitter taste, quite soluble in water and insoluble in alcohol. - 11 - Antimicrobial ..ctivity: A number of mechanisms have been postulated to account for the antimicrobial activity of streptomycin. Experimentally, it can inhibit (1 ) an oxalo-acetate-pynivate condensation • ... reaction carried out b y some organisms;(2 ) it can alter the permiability of the microbacteria cell membrane; (3 ) and it can cause inhibition of p o l y p e p t i d e synthesis and misreading of the genetic m e 3 3 age. This latter mechanism is currently believed to form the basis of the antimicrobial action of aminoglycosides in vivo. Streptomycin attaches to a protein of the 30s ribosome unit and inhibit its function in protein 3yntheis. Resistance: : ; Streptomycin resistance occur when a mutation alters or eliminates the binding site for the drug on the ribosome. Clinical U s e ; 9 Used in Tuberculosis, in systemic and urinary tract infections, enterococcal infection (aa a combined treatment. Adverse effects: Jtreptomycin cause allergy, vertigo and loss of balance. In large doae 3 , it exerts anephrotoxic ' effect. m m m i Chemistry: Kanamycin lias the following formula ^ 3.8 ^ 3 6^ 4 ° H ,in(1 consists of 3 -U-glucosamine, 6 - 0 -glucosamine linked to doxystreptamine. Kanamycin sulphate is a white powder which is readily soluble in water and quite stable at 20°C. op U A fry Antimicrobial a tivity: Kanamycin is bacteriaeidal in concentrations 1-lCtag/ial for many gram positive and gram negative bacteria particularly in alkaline medium. Proteins are often susceptible but p s e u d o m o n a s and streptococci are quite resistant. Kanamycin have sane antimicrobial action as streptomycin. Resistance; dtrains of staph and K.coli may develop i*esistance to kanamycin. She situation with regard to staph varies widely from region and does not seem to be related solely to the frequency with which the drug is used. Clinical a b d i c a t i o n : Kanamycin i3 a drug effective against a wide range of gran negative bacteria and has some use in the treatment of Tuberculosis. Its clinical application i3 confined to those patient3 who have serious infections due to gram negative organisms and in certain cases of T.3. iide Effects: Kanamycin cause Hypersensitivity, neuroto xicity (cranial nerve damage and curare-like effect) and iaphrotoxicity. ■J-KITAKYCIH: Gentamycitt 13 soluble in water and resulting solutions are stable weeks. Antimicrobial ac ti vit.y: Gentamycin i3 bacteriaeidal at concentrations of 0.5” 5fflg/ml. The activity is enhanced at alkaline pH. Hentamycin Gulphate lQxag/ml inhibit most strains of staph, and calif arms in vitro, fhe pacterieidial action i3 probably due to inhibition of protein synthesis, perhaps by causing a \ misreading of the M R M message, at the 30s unit of ribosome, ftaaiatance; 'Gentamycin resistance bacteria appear to alter ribosome protein. Microorganisms made resistant to gentamycin are usually cross-resistant to Kanamyciakeomycin, but tne reverse i3 not the case. Some bacteria inactivate gentamycin by phosphorylation, and this property can be transmitted by resistant transfer factors (RTF) from one bacteria species to another. Clinical a m i l e t i o n : The main indication for the use of gentamycin is the .urinary tract infection due to sensitive organisms; meningitis or septicaemia caused by gram negative organism can be successfully treated. bide effects: The chief toxic effect of Gentamycin is on the eight cranial nervd. Both vestibular and auditory branches may be affected, but the former is more susceptible. Urticaria and transient elevation of serum transaminases has been described. 3ULF0N G .IDE * fiMLdtr.y: These are generally white, odourless, bitter tasting crystalline powders which are much soluble in alkaline than acidic pH 14 Moat sulphonamide can be prepared aa sodium aalta which are moderately soluble and these are used for intravenous administration. Mlt ini era bia.1 activity: The action i3 sulp- o .amide is bacteriatatic and is neversible with the removal of the drug or in the presence of excess of Para-amino-benzoic acid. Susceptible microorganisms require extracellular para-amino-benzoic acid to form folic acid, an essential step in the production of purines. Sulphonaaide can enter into the reaction in place of PABA, compete for t e enzyme involved, and form non function analogues of folic acid. A3 a result, further growth of the microorganism is prevented. Resistance: Animal cells are unable to synthesis folic acid but depend on exogeneous sources and for this reason are not susceptible to sulphonamide action. Other cells which p m co a large excess of PABA are resistant to sulphonamide; and still others may actually destroy sulphonamide. Sulphonamide resistant mutants occur in most susceptible bacteria populations and tend to emmerge under suitable selected pressure The wide-spread therapeutic use of sulphonamide against gonorrhoea has resulted in the establishment of sulphonaciide resistant strains. Most recently, sulphonamide resistant meningococci have appeared. It should be specifically mentioned that rickettsiae are not only inhibited by sulphonaaide but are actually stimulated in their growth. Clinical U s e : The number of antibacterial agents is now so great that sulphonamide are first choice in lew situation. Kiningococcil infection being the clearest indicator for their U3e. This includes those forms of diseases with acute and chronic septicaemia, whether meningitis is present or not. In epidemics these drugs may be used prophylactically. In circumstances where the choice of agent is limited, usually because of cost, 3ulphonamide continue to have an important role, a3 they possess the great advantage of cheapness and ease of administration. ’ . - 16 EXPMIilEH? Materials: X. (1) Nutrient Agar, (oxoid Ltd.) - l.Og. Yeast Extract (oxoid L20) - 2•Og . Peptone (oxoid L37) - 5.0g. Naci - 5.0g. Agar No.3 (oxvoid L13) - 15.Og. Lab. leraco powder (oxoid L 29) pH (7.4) Dio tilled water to lOOOnil. Method of Preparation:14.Og of Nutrient agar was weighed and boiled in 500ml. distilled water with stirring to dissolve. then autoclaved at 121°c for 15 minutes. The solution was After which it was removed covered and kept in the regrigerator until required. (2) Gram stain - composed of: (i) Methyl violet - solution (ii) Iodine solution - 2 g KI and Ig Iodine. (iii) Safranin (iv) 0.5g in 100ml water - 0.25g in 100ml water Alcohol 955® Preparations: (i) Me thjrl violet solution: 0.25i of methylviolet was dissolved in 50ml of distilled water, poured into reagent bottles and stored in a cool place. (ii) Potassium iodide Solution: lg of Potassium Iodide and 0.5g of Iodine were mixed and dissolved in Idol, of distilled water. Volume was made to 50ml. with distilled water and packed in reagent bottles. ^ Oaframin Solution; 0.25g of safranin was ground in a morto'r with 5.0ml. of 95$ ethanol and volume made to 50m l « with distrilled water, and packed in reagent bottles. A p p a ra tu s : (1 ) Glass - Petri dishes and Reagent bottles. (2 ) Cotton wool and applicator sticks. (3) (4) Beaters and 3tirring rods. ■ :w .4.4 >iHs9 *■*&. ~ Bunsen burner. (5) Flasks and flask brushes. (6 ) Microscope and Mcro-slides. (7) Autoclave. (8 ) Oven. (9) Refrigerator. -r«3F*d- Preparation of Cotton Arabs: These were prepared from ootton sticks. ol and applicator Were then double wrapped in seta of threes and autoclaved at 115°C for 30 minutes. After the allocated times were removed and stored in a cupboard. - d-S - The glasa-petri-dishes were washed with detergent, rinsed with distilled water and dried in a hot-air-oven. These were then wrapped in autoclaving paper packets and sterilized in an autoclave at 121°G for 15 min. 'These can be catergorised into 3 sections: (i) isolation of the microorganisms and colony counting (ii) Identification. (iii) Sensitivity testing. (1) Isolation The microorganisms under test, being skin flora were collected from throe sites of body surface i.e. (i ) Pore-head (ii) The arm (iii) The leg. Using sterile cotton-swabs soaked in normal saline, swabs were made from the three parts of the body. Inoculated onto sterile agen plates which have been marked with three crossing lines. The three sections were designated with the following letters H for head A for Arm L for Leg The above plates were then incubated for 46 hours in an incubator at 37 °C. 10 19 (1) -Jo lon.v Count i n **: , Alter incubation, discrete colonies were* obtained. These were counted using "surface colony count*1 technique. (2) Identxxication: This was done under the following aubheadin ;ss- (a) (a) Gram-staining and Microscopy (b) Goagulaae test (c) Oatalase Test Gran-staining and Mi erascony Smears of different colonies from the three sections of the ager plate were aade outo microscope-slides. and then flame fixed. These were air dried After which the following staining technique was done:Slides were stained with Methyl violet solution for 20 seconds. Then washed off with tap-water and stained with Iodine solution for one minute. The iodine solution was washed off with 95h alcohol, leaving the alcohol on the slide for a few seconds. was done wit; Counter staining 3afranin and left for 20 seconds The gran-stained slides containing smears of different colonies were identified under an oilimiaer3ion lens. 10 m Slide-coagulese test was done Culture from one colony was emulsified in a drop of water on a slide. This bacterial suspension was then st*rjpe4 with a drop of plasma. Then observed under a mi roscope for the presen t or absence of coagulation. Catalase testingt A drop of 30j& hydrogen peroxide was placed on a slide smeared with microorganisms, tvA&ition of gas or effervescence was recorded* After the above identification tests, subcutures of the different colonies were done in nutrient broth. Then incubated at 37°C for 4d hours in an incubator. ■ Sensitivity testing: Standardized commercial r.ast-rings KGL 2/2 were used. These contained known amounts of antimicrobial agents. Outo sterile agar plates seeded with the microorganisms were placed the mast-rings. a 37 2 far 24 hours. surround the rings. The plates were then incubated at Upon incubation clear oonee wehe seen to Zones of inhibition ware measured in — j* 21 - - uQ. OF' GJLJNIto fHcAD) CJAGULASE TEST P la te Ha 1 Q Y 8 y S 0 Sf m MM* 13 1 IS IQ ii + + * .... ■ 23 2 4 6 6 16 B C w + - - + - - * - - - - - J i -W. *•: rM + - :.v 3 1 2 2 10 20 * 4* j ' ; 4 1 1 10 3 30 16 1* S a 'i . S' 22 ■ 3' *4-'* e 5 4 .. • ■ 6 2 ■ + 4* | + «*» - + + + - - | 4* + + - - + • - - 4- s- - a 0 - - ■- 1 • 30 • .. . 4 .. 2 . * e 2 ■ 8 9 10 3fi i? 3 10 «■» 20 10 1 20 c 4* 4 10 o - + • - 12 - mm 10 20 D □ a ~ ~ 2 2 10 20 + + + - - 1 2 5 + a +• - - 13 3 14 30 IS b X - 10 20 + + a - - 16 - •m- - 10 ? a a ◦ - - 1? - mm * 4 10 0 a 0 - - 10 - - - 20 30 D a a - - 22 aufcaEB of q o l o n iis Plate No. Y 1 2 s' *? - Iflps;.) iuagulase te s t 0 R C W Y 0 R 10 3 10 J2 0 + + •f - a 2 3 2 + + - - 3 5 o o 0 - - 6 3 0 s rj o - 3 + “ t+ o ~ J - f - 4 1 5 a* - [ - j s 13 + • 6 9 14 - ? 10 + 1 2 14 |l f 14 f 3 i r 4- + : + - a 20 - - 9 ' 20 + 0 0 - - 9 3 1 1 IS S 4- + l + - mm 10 3 2 * 6 10 0 + \ + — mm 12 ^ - - 20 j 30 o o a - 2 1 8 18 o + -t- - 13 14 20 4 2 3 j 6 + + + - - 13 - 4 - 9 10 o ■+ o - - 16 4 4 - 4 •20 J_____ o + a - - 1? 4 1 - 5 IS •f. a ~ | p 23 hu^oer uf polonies Plate I No Y 1 R 0 ir 5 ( lpg) cqagul&*~ tvqt ; C w 10 $ Y 0 n C ff + + + - W — " 2 22 ' 3 - 1 3 ; 4 5 £ 4 5 ; ■> 7 ia + 0 0 • 10 20 o + + - *“2 10 e + + + *• - ' id IE + 0 - - 20 + + 4- «*► - 7 * 10 + + ! + " 9 ro + + ] + mm: ' 10 10 + + 4* - 9 a ♦ o - $0 o 0 0 is 0 + o 18 10 + •f L9 r < o 30 i a 0 a .0 C 3 ° 0 >* O f - 6 , H ■n -- . --*U-■ 6 ? 1 1Q 8 10 9 ■*W 10 * 2 12 - «•» mm 0 - 2 3 ‘ 13 - 13 - IS 3 i "T“ * 6 2 “s ' 20 '10 18 2 2 - a 16 IS e _ IQ mm 7 1? 1 - - - - - i------Key : + indicates positiva coagulase test negative a absence af organisms 'V > GRAM - PIAIR ( dhteSn :<•*«a***; Plats Ho T Y 0 0 H f.. 1 ... 0 *w ....... 1 + + +‘ + 4- 2 + + + + + 3 + ~ 4-r j+ + 4 + + + + + 5 + + 4* 4- 6 + + + + + + 1+ 1+ 8 4" + 9 + 10 0 0 0 4- 5* 12 o 0 j0 + : 13 +c> + + + + 14 + 0 + + + 4* + o + + 16 0 0 o + + 17 o 0 0 4*, + 18 otd. 0 0 0 + + 7 + + I 15 + J._____ 0 + Y I+ [♦ + 1+ 26 ■JILi,-. - .iTAlli ( L Jl>) T7 ------ ~ Y ^ 0 j ' ^ > 1 V. Plate No 1 ' V) a c W + + 4 + >*• 2 + 0 0 4- + 3 00 + 4 + + 4 4 4* 4 5 4 + 0 6 4- 4 + + + + 4 + 4- 4- + + 4- .. ' 7 + + + 1. 8 + + 4- 9 + + + 10 O 4* 0 12 o 0 13 0 4- 0 4- 4- 14 4* 4 + + 4- 4- 15 4 + 0 + 4- 16 0 o 0 + + 17 0 0 0 4 4- KBY 4- + 4- 4- — -4 — X + indicates positive £ram stain ” negative *----- * o " no organisms % V o J) L* o 3v_> » cy "j H fX7'-' u '2 < c. -> <? -> ^ s S > _? * * J 2 £ | &* ? 3 o> 3 ■-<: ^ 2 ^ & - ti I ! <s-r t a </) X d I V 4r £_§ £ % j££.| d * ^ * -> * 4 I ( ^ I ! x I r<“ id d -ci o< ^ < ^ 0 ^ i C vTl r* O H \/i O if) G 15 o cj v/) o VJ r o'* • '1 n </\ '■— I ! ^ £ * x - T >- X 3 >- i-5 < o> o £ O <: V <■ -P c a<— <> £ £3 ^ S- t j 5 £ v' o O , t i dT ^ i i l > ^ < r i H v i n~ to* d r- </\ ‘Yi o <v? in cS o c< 29 ji ;ou>.;ion Aim con jlu~>ion Results obtained from Gram-stain, and coagulase tests complied with the ones found for staphyloccoci. particular staph, aureus as these were positive. Prom sensitivity In found to be eoagulaae testing results, it was found that the microorganisms wore most sensitive to Gentamyein and least sensitive to sulphafurazole. However, because many chemotherapeutic agent3 were used, the results can be represented as followss- GEIfTAMYCIN most- sftasitive KANAMYC1M STREPTOMYCIN order of CHLORAMPHENICOL decreasing AMPICILLIN sensitivity TETRACYCLIN CO—TRIMAZOLE SULPHAFURAZOLE (least sensitive) The above results refer only to specimens collected in Kenyatta National Hospital. Resistance to the commonly used antibacterial agents like Ampicillin, tetracycline and chloramphenicol could be due t o : (i) Development of a resistant staphylococcal strain. (ii5 Degradation of the antibacterial agents by the microorganisms. p (iii) People from whom the specimens were collected, had been in contact with.these antibacterial agents, therefore the microorganisms might have acquired resistance. Although Uentamycin has been found to be more effective against staph aureus; it should not be taken as the drug of choice. This i3 because it has a drastic 3ide effect; it affects the vestibular branch of the 8th cranial nerve. . However, auditory damage is rare. Hence, other medications should be used as first line of treatment in staphylococci skin infection. These include using hot water for cleaning body surfaces (i.e. face, a m etc.) The heat or steam will cause the staphylococci infected pores to rupture and therefore degrade the organisms. Use of soaps and antiseptics is also of importance a3 some of these are bactericidal. Antibiotics should be given as the last line of medication. 3a R 3 P E R 3 n 0 E J 1. Bailey W.R. in "Diagnostic Microbiology", 2nd Edn. P IS, 22, 31 (1966). 2. Churchill L. in "Medical Microbiology", 13th Edn, p. 236 - 255 Vol.1 (1978). 3. Collins C.H. & Lyne P.M., "Microbiological Methods 4th Edn., p. 1 0 9 . t*. 4. Oruickshank R. in "Medical Microbiology", 12th Edn., pg. 5. 236 - 255 (1973). Davis, Dulbecco, Eisen, G .rf. in "Microbiology" P 21, 122 301 (1970). 6. Ernest J., Joseph L.M.; Edward A.A. in "Medical Microbiology" p 161 (1970). 7. Frederick H.M; Ernest J; Alan G. in "Medical Pharmacology" p 467, 475 494 (197©). 8. Hillas 3. in "Antibiotics in clinical practice", p 20, 20 61* 86 (1972). 9. Jawetz E . , in "Review of Medical Microbiology" 13th Edn., p 122. (1978). 10. Lawrence P.G., Prancia 0. "Antibiotic and Chemotherapy" p 70, £8 115 147 (1971). 11. Meynell M. in "Theory and Practice in Experimental Bacteriology", p 35, 125, (1930). 12. Myruik P.W. in "Fundamentals of Medical Bacteriology and Mycology" p 153 (1974). a '’"'ersity o f l IX*A »v *--**