The increasing prevalence of penicillin-resistant Streptococuus pneumoniae urges for fast and accurate susceptibility testing methods. This study evaluated the comparability of three commonly used techniques; disk diffusion, E-test and agar dilution, to detect penicillin susceptibility in clinical isolates of S. pneumoniae. Fifty pneumococcal isolates, obtained from patients at the University of Malaya Medical Centre, were selected to include both penicillin-susceptible strains and those that had decreased susceptibility (resistant and intermediate) to penicillin. The minimum inhibitory concentration (MIC) values of penicillin to serve as the reference was determined by the agar dilution method in which, based on the MIC breakpoints recommended by the National Committee for Clinical Laboratory Standards (NCCLS), 27 strains had decreased susceptibility to penicillin with 17 strains resistant and 10 intermediate. Comparing to the agar dilution method, oxacillin disk diffusion test detected all strains with decreased penicillin susceptibility as such while E-test showed a close agreement of susceptibility (92%) of the isolates to penicillin. This confirmed that oxacillin is a good screening test for S. pneumoniae isolates with decreased susceptibility to penicillin while E-test is very reliable for rapid and accurate detection of penicillin susceptibility.
Penicillin ; Streptococcus pneumoniae ; Dilution ; Upper case ee ; Clinical

This study was to compare the replication capacity of pneumococcal isolates (serotypes 1, 7F, 19F and 23F) with their adherence pattern to monolayer cells (A549). For standardization purposes, all isolates showed a normal growth curve in both bacteriological (THB + 0.5% yeast extract with and without 2% FBS) and cell culture media (RPMI + 2% FBS). In the former media, a shorter lag phase was observed for isolate serotypes 1 and 7F in presence of serum while in the later; growth yield was lower for all isolates with stationary phase approaching OD600 of 0.01 as compared to 1.0 in bacteriological media. In the replicative analysis at different growth phases of the isolates in cell culture media, growth capacity at 3 h post-incubation was frequently twice as that at 1 h, and that at early-log phase was frequently higher than that at mid-log phase at both post-incubation times. Adherence was frequently the least at early-log phase although the isolates were in the most active state of replication to increase the number of pneumococcal cells to adhere. At mid- and late-log phases, pneumococcal adherence was frequently higher although the replication was reduced. This study marks the potential correlation between pneumococcal growth fitness and adherence capacity whereby the later may not be superior during the early growth phase.

beta-Lactamases have been identified as the major cause of antimicrobial resistance to beta-lactam antibiotics in Escherichia coli. The activities of ampicillin-sulbactam and amoxicillin-clavulanate as well as a range of beta-lactam antibiotics were studied with 87 clinical E. coli isolates from patients of the University Malaya Medical Center using the disc diffusion technique. Susceptible, intermediate and resistant categories were established based on the diameter of zones of inhibition set by the National Committee for Clinical Laboratory Standards (NCCLS). The isolates were then classified into 6 phenotypes according to the criteria stated in the methodology: S (susceptible to all beta-lactams); TL (resistant to aminopenicillins; amoxicillin-clavulanate susceptible and susceptible or intermediate to ampicillin-sulbactam); TI (resistant to aminopenicillins and ampicillin-sulbactam; susceptible to amoxicilin-clavulanate); TH-IRT (resistant to aminopenicillins; intermediate or resistant to amoxicillin-clavulanate; resistant to ampicillin-sulbactam); ESBL (resistant to aminopenicillins and oxyimino cephalosporins; positive results with the double-disc diffusion test); and CP (resistant to aminopenicillins, beta-lactam-beta-lactamase inhibitor combinations, oxyimino cephalosporins and cephamycins). Results showed that the TL phenotype was the commonest (40.2% of the isolates) followed by S (31%), TH-IRT (16.1%), ESBL and CP (3.4% each) and TI (2.3%). One isolate showed both ESBL and CP phenotypes while two isolates were classified as inconclusive. Representatives from each phenotype were further analysed for the presence of beta-lactamases which revealed a predominance of TEM and SHV enzyme producers. PCR-SSCP analysis of the SHV gene from all the ESBL and CP isolates revealed the predominance of SHV 5-type enzyme which was concurrent with our previous studies.
Phenotype ; Extended-spectrum beta lactamase ; sultamicillin ; Escherichia coli ; Ceruloplasmin

Aims: The aim of this research is to explore the presence of multidrug-resistance (MDR) Acinetobacter baumannii strains isolated from hospitalized patients in a tertiary-care center, Subang Jaya, Selangor, Malaysia and to compare their genotypic and phenotypic characteristics. Methodology and results: Clonal relationships were determined by multilocus sequence typing (MLST) and biofilm forming ability was evaluated by using 2, 3 - bis (2 - methoxy - 4 - nitro - 5-sulfophenyl) - 5 - [(phenylamino) carbonyl] - 2H-tetrazolium hydroxide (XTT) reduction assay in microplates and Congo red agar method (CRA). Four virulence genes coding for A. baumannii pilus usher-chaperone assembly protein, csuE gene; outer membrane protein, ompA gene; biofilm poly-β-1, 6-Nacetylglucosamine (PNAG) synthesis protein, pgaA gene; and acinetobactin-mediated iron acquisition protein, bauA gene were searched for in a collection of strains. Antimicrobial resistance against 11 antibiotics were studied by broth microdilution method. Seventeen A. baumannii clinical strains were isolated and MLST showed that the strains belonged to 5 distinct sequence types (STs), namely, ST-6, ST-265, ST-324, ST-325 and ST-432. Fiftythree percent of the strains were resistant to 4 or more antibiotics. Twelve strains produced biofilm and out of them, 4 were strong biofilm producer, besides, these strong biofilm producers were MDR strains and belongs to ST-6. In addition, all strains were ompA positive, biofilm producing strains were csuE and pgaA positive and only strong biofilm producing strains were bauA positive. Conclusion, significance and impact study: Our study demonstrates that the ST-6 strains in Malaysia could represent MDR, capable of forming strong biofilm and possess csuE, ompA, pgaA and bauA genes, virulence characteristics that probably help the bacteria to persist and cause infection.
Acinetobacter baumannii