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Die isolasie Aochmdriv- f ~PIPPD~~PI' ,& & &l,&-.& is. ~ e r h a a l d a ~ ~ ~ 5 0 * i ~ & # l 7 y l l r b l ~ d d gehad. In k Ld M W m tebiadhradSrr*Ilrdb#i--dis- Mitochondr 'QtrsOer cDdk &B rirerb.dsrb fd& h @E). D;, * ~ ~ ~ @ t & u L 1 ~ ~ . ~ ~ 7 4 KE/lg hiusobio~lcdOarsr.OmIE/tLrcrccbd.o~ funksieenem(PguWQLrnooQt-
616 SAMJ VOL 80 7 DES 1991 V ~ ~ w d e r s o e k e eM J n de Kock* ~ and ~ L S van Niekerk daarvan het daamp gedui dat m I& die tienew geen -d& d bedYn hrssen die Lmtrde preparate van& die argief en die snitte wat na t l7 jaar v is nie. In gevalvulhM9(maagmetulhrs)waseg~erduidelilregevaxderdeordolhe ttxmwdgDieniewge&h&~oldersoel,het bogenoemde histdogiese resultate Etlilre ubadhlb strnldme km herken word. Origens toon da die beeM van mm~ale . . dq&mdd,behalarcingeval~nM&.lndieimm~ ondersoeke is die e r t met 'n reeks adiggame getoets rnaar~enlrelehartspierseIlehet~virvirnen~n getoets. ~dieligvaubogememdeondersoeke,kandie~~ekiriag gem& word dat die -0ses van P&C seeder twyfel 'n voortreflilre metode van natmusmm-lllollsferbewadqg k, wat die pataloogtotoitstekende~we~dogiesewdersoekeinstaat steL Bagewemde redtate is anielr aangesien hierdie bea ' etode op hierdie stadium, d.ws na feitlik 2 de- nog nie geevalneer is nie. CHARACTERIZATION OF BETA-LACI'AMASES FROM CLINICAL ISOLATES OF PSEUDOMONAS M M Theron* and M J de Koek Dept. of Medical Microbiology, UOFS, Bloemfontein 9300 Resistance in Pseudornonas to B-lactam antibiotics may be attributed to various mechanisms. one of which is the action Crude 5-ladamases were extracted from Pseudomonas isolates after cells were disrupted with a French Press. Semi quantitative substrate profiles for 16 &lactan antibiotics were determined by the van de Klundert procedure for both standard reference lactamases and Rlactamases from clinical isolates. The native MW of Blactamases were estimated by Polyacrylamide Gel Electro horesis (in the absence of SDS and denaturing conditions7 and an iodine- starch-penicillin test. The native MWs of crude enzymes were compared with their substrate profiles to detirrnine if any rklationship existed between MW and substrate ~rofile. Fourteen substrate profile soups were distindshed. Under the conditions-of thewassay the appareG native MW of the unknown B-lactamases varied between 19 000 and 400 000 when compared with polypeptide reference standards. This indicates that most B-lactamases were relatively large molecules or more likely aggregates which varied in both apparent MW and affinity for different B-lactams. No clear direct relationship exist between MW and substrate profiles. This may be due to the inability of the iodine-starch-penicillin test used to estimate MW in the polyacrylamide gel electrophoresis procedure to detect enzymes primarily active against cephalosporins so that the detected ennrmes did not fully reflect the &lactarn inactivating ,capability of t6e different isolates. Alternatively, the detection limits of the van de Klundert techaique rrhy be much lower than that of the iodine-starchpenicillin procedure, -resulting in the observed disparity between the two data sets. PHENOTYPIC CHARACTERIZATION AND CELLULAR FA'ITY ACID COMPOSITION OF ACINETOBACTER SPECIES FROM CLINICAL ISOLATES Department of Medical Microbiology, UOFS Bloemfontein 9300 A total of 181 Acinetobacter isolates from clinical sources were identified by biochemical, physical and cellular fatty acid-profile procedures. Fatty acid profiles were obtained by gas liquid chromatographic analysis (GLC). The biochemical tests were based on the API.20 NE system and additional tests. Physical tests comprised growth tests at 37, 41 and 44°C. Principal component analysis of fatty acid profiles revealed two main groups when clustered by a 2-D plot program. Each main group contained two subgroups. Acinetobacter baumannii and Acinetobacter calmaceticus are included in group A and Acinetobacter iohnsonii and hybridization group 9 of Bouvet and Grirnont in group B. The currently accepted physical and biochemical tests for delineation of species do not distinguish between fatty acid profile groups. There is no correlation between tlie ability af isolates to grow at 44°C (Bowet and Grimont) and their species identification by either the API 20 NE system or the microbial identification system (MIDI). based on cellular fatty acid composition. Biochemical and physical tests remain of limited value in identiwg strains at the species level and are of no value for epidemiological strain tracking. GLC proved to be a valuable tool both for species identification and strain tracking in the hospital environment. THE PLACE OF PENICILLIN THERAPY FOR GONOCOCCAL INFECTION H+I W1 P L Botha* B de R i d and M M Anthony* *Medical Microbiology University of the Orange Free state Wealth Department, Municipality Bloemfontein Background: The validity of penicillin therapy in the empirical treatment of purulent ure- discharge is questioned in the light of worldwide evidence that there is a disquieting decrease in the sensitivity of Neisseria gonorrhoeae to this antibiotic. Traditionally gonococci are exquisitely sensitive (MIC < 0,01 mg/l) to benzyl penicillin: Aim: An investigation into the current status of penicillin as first line treatment of gonococcal infection in Bloemfontein was undertaken to ensure effective therapy. Materials and Methods: Urine specimens were collected from male patients attending a municipal sexually transmitted disease clinic. Susceptibility of N. gonorrhoeae isolates to penicillin and a Chydroxy quinolone was determined by disc diffusion and minimal inhibitory concentration niethods, Results: Full penicillin sensitivity was observed in - 6% of isolates. Beta-lactamase activity was demonstrated in 14% and the MIC 90 was 4 mg/l. Marginal differences in enicillin sensitivity are not demonstrable by the disc d& sion test.
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