Detection of Pathogenic Leptospira in Rats and Phylogenetic Analysis Using Outer Membrane Lipoprotein Lipl32 Gene at Two Major Public Markets

@#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.

Distribution of virulence genes and the molecular epidemiology of Streptococcus pyogenes clinical isolates by emm and multilocus sequence typing methods

@#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.

Molecular Detection of Cryptococcus from Formalin-fixed Paraffin-embedded Tissue Sample - A Noncultural Approach for Diagnosis of Primary Pulmonary Cryptococcosis

@#Despite pulmonary cryptococcosis showing good response to antifungal treatment, making accurate diagnosis in a timely manner remains a challenge. Tissue for fungal culture is less sensitive, nonspecific, and time-consuming to allow for prompt diagnosis. We herein report a case of 31-year-old woman with no known medical illness but presented with history of cough and haemoptysis for two months. The chest X-ray and bronchoscopy revealed left lung mass. While tuberculosis showed negative result and no growth observed in the tissue culture, the histopathological examination (HPE) finding was suggestive of fungal infection. The formalin-fixed paraffin-embedded (FFPE) tissue was sent for molecular testing, which revealed Cryptococcus neoformans. This report emphasises on the advantages and limitations of polymerase chain reaction (PCR) as an alternative method to confirm the diagnosis in cases of culture-negative fungal infection.

Suppurative BCG Lymphadenitis and the Importance of Microbiology Investigations – A Case Report

@#Suppurative BCG lymphadenitis can easily be overlooked, as it mimics other diseases such as tuberculous lymphadenitis. A case of a three-month old female infant who received the BCG vaccination at birth presented with isolated left axillary mass at two months of age. She was initially treated as lymph node abscess but was referred to the hospital due to the increasing size of the swelling. Needle aspiration was done and the microbiology analysis came out positive for acid-fast bacilli. She was planned for syrup isoniazid; however, the management team withheld treatment until they were certain of the identity of the bacteria. The bacteria was confirmed by the molecular method to be Mycobacterium bovis BCG strain. The case report highlights the importance of the microbiology investigations for appropriate management in this case.

A Fatal Case of Rhinocerebral Mucormycosis

@#Rhinocerebral mucormycosis is a potentially fatal and progressive angioinvasive fungal infection. It is classically described in patients with uncontrolled diabetes mellitus and hematological malignancies. This report describes a case of progressive rhinocerebral mucormycosis in a patient with poorly controlled diabetes who was on prolonged prednisolone therapy for autoimmune kidney disease. The patient, who was a female, presented to hospital with headache, orbital pain and nasal bridge swelling. Black eschar on nasal mucosae was present on admission. Later, she was started on intravenous fluconazole for the diagnosis of fungal sinusitis. Subsequently, she developed intracerebral haemorrhage complicated with transtentorial herniation. Diagnosis of rhinocerebral mucormycosis was later observed by a laboratory finding and the treatment was changed to intravenous amphotericin B. However, the patient succumbed to her illness on the 6th day of hospitalisation. This report discusses the risk factors associated with rhinocerebral mucormycosis as well as the underlying pathogenesis. This report will also highlight the importance of early diagnosis and appropriate treatment for mucormycosis to improve prognosis in patients.

False Positive Blood Culture From Automated Microbial Detection System in Severe Malaria – A Case Report

@#Automated microbial detection system (AMDs) are design to detect early growth of bacterial and fungal. We herein report a rare case of false positive blood culture by AMDs in Plasmodium falciparum infection. A 41-year-old previously healthy lady, with recent history of travelling to Lagos, Nigeria had presented to the casualty with history of fever and lethargy for three days. There was no malaria prophylaxis taken prior to the travelling history. Peripheral blood smear confirmed the presence of young trophozoite of Plasmodium falciparum with parasitemia of 7%. Concurrent blood culture sent was positive, however all subcultures were negative for any growth. She was treated with intravenous artesunate however succumbed to death on the day of admission due to severe falciparum infection complicated with multiorgan failure and shock. The aim of this report is to highlight, the circumstances that can trigger the false positive AMDs detection and the possible underlying mechanism.

Species distribution and clinical profiles of coagulasenegative staphylococci (CoNS) isolated from blood cultures among paediatric patients in Hospital Kuala Lumpur

@#Introduction: Coagulase-negative staphylococci (CoNS) is often considered as a culture contaminants but it can potentially be pathogenic to patients with risk factors. A combination of species identification and clinical criteria has been suggested in determining true CoNS bacteraemia. Objectives: To identify the species distribution, antibiotic susceptibility patterns and clinical profiles of CoNS isolated from blood cultures among paediatric patients in Hospital Kuala Lumpur (HKL). Methods: This study involved CoNS isolation from blood cultures of paediatric in-patients of the Paediatric Institute HKL. Isolates were identified to species level using Analytical Profile Index Staph identification strips and antimicrobial susceptibility pattern following Kirby-Bauer Disc Diffusion method. The clinical profiles of patients were obtained from their medical records. Results: Eleven CoNS species were identified from 148 isolates. Staphylococcus epidermidis was the most frequent species isolated (67.6%). The majority of the isolates showed resistance to penicillin (85.8%); while 70.3% were methicillin-resistant (MR) CoNS, which demonstrated a significant association with true infection (p=0.021). Predictors for significant CoNS infection included thrombocytopaenia, presence of predisposing factors, nosocomial infection, blood collected from peripheral vein, and CoNS isolated from two consecutive blood cultures. The most common predisposing factors for the isolation of CoNS were the presence of peripheral (54.1%) and central venous catheters (35.1%). Conclusion: CoNS can cause significant bloodstream infections. The isolation of CoNS from blood cultures should be carefully interpreted by considering the predictive factors. Local data regarding predictive factors of patients with culture-positive CoNS, species distribution and antimicrobial susceptibility pattern are useful to determine the significance of blood culture results and care management of patients

Serotype 15C Streptococcus pneumoniae with Third Cranial Nerve Palsy: Unusual Serotype and Presentation

@#We present a unique case of pneumococcal meningitis caused by serotype 15C, a non-vaccine serotype, which is long thought to be non-virulent. Our patient is a young lady with a known case of plaque psoriasis, presented with sudden onset of left oculomotor nerve palsy and severe headache two days prior to admission. The clinical features were initially mimicking of brain tumour and brain aneurysm. The diagnosis of pneumococcal meningitis was confirmed by the bacterial antigen test and genome detection using multiplex PCR from the CSF. The blood culture also grew Streptococcus pneumoniae. Serotyping was performed on the isolate using Neufeld's Quellung method and it was identified as serotype 15C. Psoriatic skin disease was identified as a potential source of this invasive infection.

The Prevalence of Soil-Transmitted Helminths Infection and Its Association with Anaemia Among Refugee School Children in The Klang Valley, Malaysia

@#Introduction: Soil-transmitted helminths (STHs) are considered one of the neglected parasitic diseases that cause major health problems and other related complications. The aim of this study was to determine the prevalence of STHs and anaemia among school children of little-known refugee communities in Malaysia. Methods: A cross-sectional study was conducted among 148 refugee school children in the Klang Valley, Malaysia, from 2017 to 2019. Data were collected using a questionnaire, in addition to haemoglobin testing while faecal samples were screened for STHs eggs using the formalin-ether concentration technique. Results: The result showed that the prevalence of STHs in this study was 37.2%. Trichuris trichiura infection was the most prevalent (46.4%) followed by Ascaris lumbricoides (44.9%) and hookworms (8.7%). About 37.8 % of the respondents were found to be anaemic. Binary logistic regression showed significant infection with STHs complicated with anaemia (AOR: 3.67, 95 % CI 1.595-8.5). Conclusion: The evidence from this study suggests proper and strategic interventions to reduce morbidity. An alternative and efficient system is needed to provide the schoolchildren with basic care for sanitation and health provision. It is recommended that voluntary clinics provided by non-government organisations focus on counselling and advice on personal hygiene of the children.

Candida isolates from pregnant women and their antifungal susceptibility in a Malaysian tertiary-care hospital

Objective: Pregnant women are susceptible to vaginal colonization and infection by yeast. The purpose of the study was to determine the prevalence of Candida spp in high vaginal swabs of pregnant women and their antifungal susceptibility. Methods: High vaginal swab samples received from Serdang Hospital, Selangor, Malaysia during 2011 initially had microscopic examination, Gram-staining and fungal culture. These were finally confirmed by growth in chromogenic medium [CHROMagarCandida; Difco BBL, USA] and commercial biochemical identification kit [API 20C AUX; bioMérieux, Lyon, France]. Antifungal susceptibility was performed by E-test method. Results: Out of 1163 specimens 200 [17.2%] candida spp were confirmed from high vaginal swabs of pregnant women. Candida albicans [83.5%] is the most common species detected followed by Candida glabrata [16%] and Candida famata [0.05%]. All C.albicans and C.famata isolates were susceptible to fluconazole while C.glabrata isolates were dose dependent susceptibility. First and second trimester, and diabetes were considered significant factors in patients for the vaginal candidiasis [p < 0.001]. Conclusions: In pregnant women, C. albicans was the frequently isolated yeast from high vaginal swabs. Routine screening and treatment are important of pregnant women regardless of symptoms

Prevalence of macrolide resistance and in vitro activities of six antimicrobial agents against clinical isolates of Streptococcus pneumoniae from a multi-center surveillance in Malaysia

The in vitro activities of 6 antimicrobial agents against clinical isolates of Streptococcus pneumoniae (pneumococci) were investigated and the erythromycin minimum inhibitory concentrations (MICs) were correlated with the two major macrolide resistance determinants, mef(A) and erm(B). MICs of commonly used antibiotics as well as the presence of macrolide resistance determinant genes in all isolates were tested. Seventy one pneumococcal isolates collected at Institute for Medical Research (IMR) were included in this study. Phenotypic characterization, MIC determination using E-test strips and polymerase chain reactions for antibiotic resistance determination were included. Among the isolates, 25 (35.2%) isolates were erythromycin susceptible, 3 (4.2%) were intermediate and 42 (60.6%) were resistant. Fifty three isolates (74.7%) were found with mef(A) alone, 15 (21.1%) isolates with erm(B) + mef(A) combination and 3 (4.2%) isolates with none of the two genes. The in vitro activity of penicillin, amoxicillin clavulanic acid, ceftriaxone and cefotaxime is superior to trimethoprim-sulfamethoxazole and erythromycin. In conclusion, pneumococcal isolates in this study were highly susceptible to penicillin with very low MICs. However, a very high prevalence rate of erythromycin resistance was observed. Erythromycin resistant S.pneumoniae isolates with both mef(A) and erm(B) showed very high MICs ≥256 μg/mL.