Introduction: Limited studies have been documented on the presence of pathogenic Leptospira in public markets serving the community in sub-districts of Selangor. The aim of this study was to detect the presence of pathogenic Leptospira in rats using a gene encoding an outer membrane lipoprotein LipL32. Methods: Polymerase chain reaction (PCR) was performed using LipL32 primers on sixty kidney samples of rats trapped at two locations of study; Pasar Borong Selangor in Seri Kembangan and Pasar Basah Bandar Baru Bangi in Bangi. Results: Out of 60 samples analysed, 36.7% were positive for the presence of LipL32. All positive samples highly matched (>94%) nucleotide sequence for LipL32 of pathogenic Leptospira and related to the pathogens through phylogenetic analysis. Conclusion: The detection of LipL32 indicates the potential presence of pathogenic Leptospira species at public markets. Although only 60 rats were successfully trapped, the rats are mobile and might further transmit the pathogenic organisms to other areas.

Background: Streptococcus pyogenes has a variety of virulence factors and the predominant invasive strains differ according to specific emm types and geographical orientation. Although emm typing is commonly used as the gold standard method for the molecular characterisation, multilocus sequence typing (MLST) has become an important tool for comparing the genetic profiles globally. This study aimed to screen selected virulence genes from invasive and non-invasive clinical samples and to characterise the molecular epidemiology by emm typing and MLST methods. Materials and Methods: A total of 42 S. pyogenes isolates from invasive and non-invasive samples collected from two different tertiary hospitals were investigated for the distribution of virulence factors and their molecular epidemiology by emm and multilocus sequence typing methods. Detection of five virulence genes (speA, speB, speJ, ssa and sdaB) was performed using multiplex polymerase chain reaction (PCR) using the standard primers and established protocol. Phylogenetic tree branches were constructed from sequence analysis utilised by neighbour joining method generated from seven housekeeping genes using MEGA X software. Results: Multiplex PCR analysis revealed that sdaB/speF (78.6%) and speB (61.9%) were the predominant virulence genes. Regardless of the type of invasiveness, diverse distribution of emm types/subtypes was noted which comprised of 27 different emm types/subtypes. The predominant emm types/subtypes were emm63 and emm18 with each gene accounted for 11.8% whereas 12% for each gene was noted for emm28, emm97.4 and emm91. The MLST revealed that the main sequence type (ST) in invasive samples was ST402 (17.7%) while ST473 and ST318 (12% for each ST) were the major types in non-invasive samples. Out of 18 virulotypes, Virulotype A (five genes, 55.6%) and Virulotype B (two genes, 27.8%) were the major virulotypes found in this study. Phylogenetic analysis indicated the presence of seven different clusters of S. pyogenes. Interestingly, Cluster VI showed that selected emm/ST types such as emm71/ST318 (n=2), emm70.1/ST318 (n=1), emm44/ST31 (n=1) and emm18/ST442 (n=1) have clustered within a common group (Virulotype A) for both hospitals studied. Conclusion: The present study showed that group A streptococcci (GAS) are genetically diverse and possess virulence genes regardless of their invasiveness. Majority of the GAS exhibited no restricted pattern of virulotypes except for a few distinct clusters. Therefore, it can be concluded that virulotyping is partially useful for characterising a heterogeneous population of GAS in hospitals.

Aims: Acinetobacter baumannii has been identified as one of the six most pathogenic bacteria that is the cause of most hospital bacterial infections according to Infectious Disease Society of America (IDSA). These nosocomial pathogens are notorious worldwide due to its ability in causing lethal infections among immunocompromised patients and its resistance to many strong antibiotics. This study aims to compare the expressed proteins of two A. baumannii strain, ATCC 19606 and a pathogenic clinically isolated strain known as AB-13. Methodology and results: AB-13 clinically strain was isolated from the lower respiratory tract of a patient with pneumonia. In this study, the proteomic profile of both ATCC 19606 and AB-13 are produced using 2-dimensional gel electrophoresis. The total protein contents were extracted, quantified and separated using 2-DE with a pH range of 4-7 to acquire the proteomic profile for comparison. The final analytical gel was analysed using Delta2D software and among the 324 protein spots successfully resolved, 10 spots exhibited signs of differential expression with 7 spots found to be downregulated and 3 spots upregulated (p< 0.01). These differences could signify the evolution AB-13 has undergone as it acquires traits ultimately aiding in its survivability, antimicrobial resistance and pathogenicity within varied environments especially during infections. Conclusion, significance and impact of study These findings support the presence of variation in AB-13 from a proteomic perspective, highlighting the pathogen’s evolution improving survivability and pathogenicity, warranting in-depth exploration towards understanding A. baumannii virulence and pathogenicity.
Acinetobacter baumannii--pathogenicity ; Two-Dimensional Difference Gel Electrophoresis ; Proteomics

Introduction: Methicillin-Resistance Staphylococcus aureus (MRSA) is known as a major nosocomial pathogen in healthcare. However, it has now spread in the community known as community-acquired MRSA (CA-MRSA). Thus, the survival and pathogenicity of CA-MRSA isolates were assessed using in vivo peritonitis model with comparison to ATCC-MRSA. Two CA-MRSA isolates; CA-MRSA1 and CA-MRSA2 that were isolated from healthy population, were studied and compared. Methods: Mice were assigned into 4 groups and injected intraperitoneally with ATCC-MRSA, CA-MRSA1 or CA-MRSA2, respectively. Sterile Dulbecco’s Phosphate-Buffered Saline (DPBS) represents negative control. Mice were observed twice daily, 0-72 hours of post-infection. Any signs of distress were recorded for severity score and survival analyses. Mice were euthanised at 72 hours post-inoculation or by referring to the Peritonitis Severity Scoring (PSS) system. Organs of interest, peritoneal lavage and abscess were processed for bacterial counts. Tissue samples were analysed for histopathological scores. Results: All mice inoculated with MRSA showed clear signs of illness with peritonitis symptoms of p<0.001 and comparable PSS scores were recorded in all infected mice groups. Intraperitoneal injection of lethal dose of MRSA resulted in significant death of ATCC-MRSA (p<0.05) and CA-MRSA-infected mice (p<0.01), compared to the un-infected. Bacterial burden was significantly high in all samples harvested from mice challenged with CA-MRSA2 compared to ATCC-MRSA except in abscess and lung. Significant liver necrosis and spleen inflammation were observed in CA-MRSA1, and lung inflammation in ATCC-MRSA-infected mice. Conclusion: Nasal carriage CA-MRSA isolates from a healthy population has the potential to cause peritonitis with comparable severity as ATCC-MRSA.

The aim of this study was to study the genotype of a hospital collection of Group B Streptococcus (GBS) from invasive and non-invasive sites. Fifty-one pre-characterised human of GBS were re-identified and further analysed by multilocus sequence typing (MLST) in relation to previously published serotypes. Fifteen sequence types (ST) were found with ST1 being the most predominant. ST1 was also associated with majority of the invasive isolates. The genotypic distribution patterns of GBS in this study were largely in agreement with previous reports from other countries indicating the tendency of certain genotypes to prevail in human infection settings.

Abstract We describe our technique of single-bundle PCL reconstruction using a bony femoral press-fit system. 9 patients underwent PCL reconstruction using our pressfit system. The surgical procedure is described in detail. Post-operatively, 5 patients were available for assessment and review. Four patients gave a final Lysholm score of 92 – 100%. Three patients gave a Hospital for Special Surgery (HSS) score of excellent, one fair and one poor. Two patients gave a reduced Tegner activity score post-operatively while the other three gave a similar score post-operatively. Assessment using KT-1000 revealed four patients with a side-to-side difference of less than 3 mm (average side-to-side difference, 1.87 mm), while one patient exhibited a side-to-side difference of 5.8 mm. We believe that our technique enhances tunnel healing through usage of a bone-plug fixation and provides a cheap alternative for graft fixation on the femoral side in PCL reconstruction.

Aims: To characterize the genotypic distribution of mec complex, bla complex, methicillin-resistance level (cefoxitinMIC) and β-lactamase activity in carriage methicillin-resistant Staphylococcus species for a potential correlation. Methodology and results: Biochemical test, 30 µg cefoxitin diffusion disc test, cefoxitin E-test, mec and bla complexes distributions, Pbp2a and β-lactamase assays were conducted to characterize phenotypic and genotypic of MRSA and MRCoNS in our collection. Phylogenetic tree was constructed using MEGA6 software to trace the diversity of blaZ gene of MRSA and MRCoNS. Sixteen MRSA and nineteen MRCoNS were identified by biochemical tests followed by 30 µg cefoxitin antibiotic disc susceptibility test and mecA gene screening. Twenty nine isolates carry complete mecA genes (2.1 kb), incomplete mec regulator (negative or truncated) and positive Pbp2a assay for both MRSA and MRCoNS. Only MRCoNS SC177 isolate with cefoxitin MIC of 32 µg/mL carries complete mec complex. Thirty-one of thirty-five isolates carry complete bla complex (blaZ, blaRI, blaI) with 10 MRSA produce strong β-lactamase and cefoxitin MIC of ≥12 µg/mL. Only 4 MRCoNS with cefoxitin MIC of ≤8 µg/mL produce strong β-lactamase. The diversity of blaZ gene was demonstrated by phylogenetic analysis and unusual amino acid mutation at position 145 for MRSA SA60 isolate may compromise its β-lactamase activity with low cefoxitin MIC level (2 µg/mL). Conclusions, significance and impact of the study: Isolates that carry complete complete mecA gene were largely consistent with the expression of Pbp2a. Nevertheless, there is no clear correlation of mec regulator genes in relation to cefoxitin-MIC in both methicillin resistant (MR) Isolates that carry Staphylococcus species. On the other hand, various expression level of β-lactamase may correlate with cefoxitin-MIC level in MRSA as compared to MRCoNS.

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

Background: Several occupational diseases of multiple origins are encountered among abattoir workers. Presence of indicator microorganisms (coliforms) on hands of workers can be used a gauge for hygienic practices. Methods: A cross-sectional study was performed to assess the prevalence of E.coli and enterobacteriaceae among Halal abattoir workers in some government halal abattoirs of Malaysia. A total of one hundred and sixty-five hand swab samples were collected from workers of Halal abattoirs in Malaysia. The samples were subjected to microbiological analysis for characterisation and serotyping. Results: The results have shown that no Escherichia coli O157:H7 was isolated on the hands of abattoir workers before and after work. However, a total prevalence of 9.7% was recorded for all samples during work. For non-O157:H7, total prevalence of 33.3% during work and 13% after work were obtained. High prevalence was recorded in sample taken during work from Tampin, Jasin and Kemaman (100% each) while low prevalence where observed in Shah Alam, Banting and Ipoh (20% each). Conclusions: Based on the findings the hygienic practices of hand washing among the workers in few locations was found to be low especially after work.