Nabilah Ismail

Molecular Characterization of Leptospira spp. in Environmental Samples from North-Eastern Malaysia Revealed a Pathogenic Strain, Leptospira alstonii

The presence of pathogenic Leptospira spp. in the environment poses threats to human health. The aim of this study was to detect and characterize Leptospira spp. from environmental samples. A total of 144 samples comprised of 72 soil and 72 water samples were collected from markets and recreational areas in a north-eastern state in Malaysia. Samples were cultured on Ellinghausen and McCullough modified by Johnson and Harris media. Leptospires were positive in 22.9% (n = 33) of the isolates. Based on partial sequences of 16S rRNA, a pathogenic leptospire, Leptospira alstonii (n = 1/33), was identified in 3% of the isolates followed by intermediate leptospire (L. wolffii, n = 1/33, and L. licerasiae, n = 7/33) and nonpathogenic leptospire, L. meyeri (n = 22/33) in 24.2% and 66.7%, respectively. This study demonstrates the presence of a clinically significant pathogenic L. alstonii in the environments which could pose health risks to the occupants and visitors.

The prevalence and preventive measures of the respiratory illness among Malaysian pilgrims in 2013 hajj season

Abstract Background. Respiratory illness continues to exert a burden on hajj pilgrims in Makkah. The purpose of this study is to determine the prevalence of respiratory illness and its associated factors among Malaysian hajj pilgrims in 2013 and to describe its preventive measures. Methods. A cross-sectional study was conducted in Makkah and Malaysia during the 2013 hajj season. A self-administered proforma on social demographics, previous experience of hajj or umrah, smoking habits, co-morbid illness and practices of preventive measures against respiratory illness were obtained. Results. A total of 468 proforma were analysed. The prevalence of the respiratory illness was 93.4% with a subset of 78.2% fulfilled the criteria for influenza-like illness (ILI). Most of them (77.8%) had a respiratory illness of <2 weeks duration. Approximately 61.8% were administered antibiotics but only 2.1% of them had been hospitalized. Most of them acquired the infection after a brief stay at Arafat (81.2%). Vaccination coverages for influenza virus and pneumococcal disease were quite high, 65.2% and 59.4%, respectively. For other preventive measures practices, only 31.8% of them practiced good hand hygiene, ∼82.9% of pilgrims used surgical face masks, N95 face masks, dry towels, wet towels or veils as their face masks. Nearly one-half of the respondents (44.4%) took vitamins as their food supplement. Malaysian hajj pilgrims with previous experience of hajj (OR 0.24; 95% CI 0.10–0.56) or umrah (OR 0.19; 95% CI 0.07–0.52) and those who have practiced good hand hygiene (OR 0.35; 95% CI 0.16–0.79) were found to be significantly associated with lower risk of having respiratory illness. Otherwise, pilgrims who had contact with those with respiratory illness (OR 2.61; 95% CI 1.12–6.09) was associated with higher risk. Conclusions. The prevalence of respiratory illness remains high among Malaysian hajj pilgrims despite having some practices of preventive measures. All preventive measures which include hand hygiene, wearing face masks and influenza vaccination must be practiced together as bundle of care to reduce respiratory illness effectively.

Ocular surface infections in northeastern state of malaysia: a 10-year review of bacterial isolates and antimicrobial susceptibility.

Objective: Ocular surface infections that include infections of conjunctiva, adnexa, and cornea have the potential risk of causing blindness within a given population. Empirical antibiotic therapy is usually initiated based on epidemiological data of common causative agents. Thus, the aims of this study were to determine the bacterial agents and their susceptibility patterns of isolates from ocular surface specimens in our hospital. Methods: This is a retrospective analysis and records of bacterial isolates from ocular surface specimens in Hospital Universiti Sains Malaysia from January 2001 to December 2010 were examined. Specimens were processed according to standard laboratory procedures. Antimicrobial susceptibility testing was conducted based on Clinical and Laboratory Standards Institute recommendations. Only single, nonrepetitive isolates were included in the analysis. Results: A total of 1,267 isolates were obtained during the study period, which comprised Staphylococcus aureus (n = 299, 23.6%), Pseudomonas aeruginosa (n = 194, 15.3%), Streptococcus pneumoniae (n = 108, 8.5%), Haemophilus influenzae (n = 100, 7.9%), Haemophilus parainfluenzae (n = 84, 6.6%), and Enterobacter spp. (n = 81, 6.4%). Fungi contributed to 4.4% of the total isolates. The antimicrobial susceptibility testing demonstrated that gram-positive bacteria were generally resistant to gentamicin (19%–57%), whereas gram-negative bacteria were resistant to chloramphenicol (27%–58%). Conclusions: Based on the above results, knowledge of the initial Gram stain findings is imperative before the commencement of empirical antibiotic therapy. Therefore, a simple Gram staining for all eye specimens is highly recommended.

Molecular detection of leptospirosis and melioidosis co-infection: A case report

Leptospirosis and melioidosis are important tropical infections caused by Leptospira and Burkholdheria pseudomallei, respectively. As both infections share similar clinical manifestations yet require different managements, complementary laboratory tests are crucial for the diagnosis. We describe a case of Leptospira and B. pseudomallei co-infection in a diabetic 40-year-old woman with history of visit to a freshwater camping site in northern Malaysia. To our knowledge, this is the first case of such double-infection, simultaneously demonstrated by molecular approach. This case highlights the possibility of leptospirosis and melioidosis co-infections and their underlying challenges in the rapid and accurate detection of the etiologic microorganism.

Microbiological aetiology of acute dacryocystitis in hospital Universiti Sains Malaysia, Kelantan Malaysia

Abstract Objective To determine the microbiological aetiology of acute dacryocystitis presented to the Hospital University Sains Malaysia, Kubang Kerian, Kelantan in 5 years duration from 2005 until 2010. Methods This is a retrospective analysis of patients who were clinically diagnosed as acute dacryocystitis from 2005 until 2010 to determine the regional microbiological pattern. The age, gender, predisposing factors, intravenous antibiotics and their microbiological results of discharge from punctal expression were collected. The laboratory procedures were in accordance with the Clinical and Laboratory Standards Institute guidelines. Results There were 23 patients admitted to the eye ward during study period. Females (n=17) outnumbered males (n=6). Majority of isolates were Gram-positive bacteria (n=10, 43.4%) followed by Gram-negative isolates (n=2, 12.9%). The most predominant isolates were Streptococcus pneumonia (S. pneumonia) (21.7%) followed by Staphylococcus epidermidis (S. epidermidis) (13.0%). Conclusions S. pneumoniae was the commonest gram positive organism identified in our study. 47.8% patients showed resistant to initial empirical treatment.

Leptospirosis seropositivity and its serovars among cattle in Northeastern Malaysia

Background: Leptospirosis is a zoonotic disease that infects human and livestock which causes economic losses to the farmers. It has been reported as one of the causes of reproductive failure in cattle and other ruminants, determining abortions, stillbirth, weak newborns, and decrease in their growth rate and milk production. Aim: The objectives of this study were to determine the leptospirosis seroprevalence and to identify the predominant infecting serovars among cattle. Materials and Methods: A cross-sectional study involving 420 cattle from six randomly selected districts in Kelantan was conducted. A serological test using the microscopic agglutination test was conducted in the Institute of Medical Research with a cutoff titer for seropositivity of ≥1:100. Results: The overall prevalence of leptospirosis seropositivity among cattle in this study was 81.7% (95% confidence interval: 63.5, 80.1). The most common reaction obtained with the sera tested was from the serovar Sarawak with 78.8%. Conclusion: A high seroprevalence of leptospiral antibodies was found among cattle in Northeastern Malaysia. These findings urge that more studies are required to determine the reasons for the high seroprevalence among the cattle along with its transmission and pathogenicity of the local serovar Sarawak.

Leptospirosis and Workplace Environmental Risk Factors among Cattle Farmers in Northeastern Malaysia

Background: Leptospirosis is an emerging zoonosis and its occurrence has been reported to be rising globally. The environment plays an important role in the survival of Leptospira and determines the risk of infection. Those who were exposed to and had contact with contaminated environment through their occupational, recreational and other activities can be infected with the organism. Objective: To determine the seroprevalence of leptospirosis among cattle farmers, prevalence of pathogenic Leptospira, and the workplace environmental risk factors for leptospirosis among cattle farmers in northeastern Malaysia. Methods: A cross-sectional study involving 120 cattle farmers was conducted. The participants answered an interviewer-guided questionnaire that consisted of sociodemographic and workplace environment characteristics questionnaire, before having their blood sample taken for microscopic agglutination test (MAT). Seropositivity was determined using a cut-off titer of ≥1:100. 248 environmental samples were also collected from the cattle farms for polymerase chain reaction (PCR). Results: The overall seroprevalence of leptospiral antibodies was 72.5% (95% CI 63.5% to 80.1%) and the prevalence of pathogenic Leptospira in the cattle farms environment was 12.1% (95% CI 8.4% to 17.0%). The independent factors associated with seropositivity of leptospirosis among cattle farmers were positive pathogenic Leptospira in the environment (Adj OR 5.90, 95% CI 1.34 to 26.01) and presence of garbage dumping in the farm (Adj OR 2.40, 95% CI 1.02 to 5.65). Conclusion: Preventing leptospirosis incidence among cattle farmers necessitates changes in work environment. Identifying modifiable factors may also contribute to the reduction of infection.

Anti-microbial Activity of Aqueous Quercus infectoria Gall Extract against Pathogenic Leptospira

Background Quercus infectoria gall extract is known to have broad spectrum anti-microbial activity in vitro. This study was conducted to determine the anti-microbial activity of Q. infectoria gall extract against pathogenic Leptospira and to evaluate the morphological changes of extract-treated cells using a scanning electron microscope (SEM). Methods A two-fold serial microdilution broth assay was used to determine the minimum inhibitory concentration (MIC) of aqueous Q. infectoria gall extract against the L. interrogans serovar Javanica and the L. interrogans serovar Icterohaemorrhagiae, at concentrations ranging from 4.00 mg/mL to 0.0078 mg/mL. The minimum bactericidal concentration (MBC) was determined by sub-culturing the broth from the microtiter plate wells that showed no apparent growth or turbidity to the freshly prepared Ellinghausen-McCullough-Johnson-Harris (EMJH) broth media, and it was subsequently observed under a dark field microscope following three weeks of incubation for purposes of growth detection. The cell morphology of both extract-treated and untreated L. interrogans serovar Icterohaemorhagiae was analysed using the SEM. Results The results of the broth microdilution assay demonstrate that the aqueous Q. infectoria gall extract possesses anti-microbial activity against both of the L. interrogans serovars with MIC values of 0.125 mg/mL. The MBC values for the L. interrogans serovar Javanica and the L. interrogans serovar Icterohaemorhagiae are 0.125 mg/mL and 0.250 mg/mL, respectively. The SEM micrograph shows changes in shape and size of the extract-treated cells (at 8× MIC) in comparison to the untreated cells. Conclusion The Q. infectoria gall extract displays anti-microbial inhibition and killing activity against the pathogenic Leptospira isolates, and thus has the potential for further exploration of its efficacy and use in the treatment of leptospirosis.

Development and validation of pan-Leptospira Taqman qPCR for the detection of Leptospira spp. in clinical specimens

BACKGROUND Early diagnosis of leptospirosis is important for ensuring better clinical management and achieving better outcomes. Currently, serological assays suffer from inconsistent performance and are less useful for early diagnosis of leptospirosis. As an alternative, qPCR is more sensitive, specific and able to detect the presence of leptospiral DNA during the acute phase of the infection. Meanwhile, most molecular assays do not detect the non-pathogenic group of Leptospira, even though these groups may also infect humans, although less frequently and less severely. METHODS A set of primers and probe targeting rrs genes of 22 Leptospira spp. were designed and evaluated on 31 Leptospira isolates, 41 other organisms and 65 clinical samples from suspected patients. RESULTS The developed assay was able to detect as low as 20 fg Leptospira DNA per reaction (equivalent to approximately 4 copies) and showed high specificity against the tested leptospiral strains. No cross amplification was observed with the other organisms. During the evaluation of the confirmed clinical specimens, the developed assay was able to correctly identify all positive samples (n = 10/10). One amplification was observed in a negative sample (n = 1/55). The sequencing of the PCR product of the discordant sample revealed that the sequences were similar to those of L. interrogans and L. kirschneri. CONCLUSION The findings suggest that the developed Taqman qPCR assay is sensitive, specific and has potential to be applied in a larger subsequent study.

The Sensitivity, Specificity and Accuracy of Warning Signs in Predicting Severe Dengue, the Severe Dengue Prevalence and Its Associated Factors

Objectives: To study Malaysian dengue clinical practice guideline (CPG) warning signs (WS) in predicting severe dengue (SD) and its associated factors among confirmed cases presented to a teaching hospital in north-eastern Malaysia in 2014. Methods: A cross-sectional study was performed in February 2015 using secondary data acquired from the hospital records. There were 2607 confirmed dengue cases presented to Hospital Universiti Sains Malaysia (HUSM) in 2014. Seven hundred patients were selected after proportionate stratified random sampling conducted according to the number of cases in 12 different months in 2014. Data were collected and analysed using SPSS version 22.0. Results: Severe dengue outcomes represented 4.9% of cases. The prevalence of any of WS in SD was 91.2%. The most common WSs prior to SD were persistent vomiting (55.9%), and abdominal pain/tenderness (52.9%). The most sensitive warning sign in detecting SD was abdominal pain (59%). Specificity of individual WS were generally good, especially of clinical fluid accumulation (99%), hepatomegaly (98%) and mucosal bleeding (93%). Factors associated with SD were persistent vomiting (Adjusted odds ratio (aOR)): 2.41), mucosal bleeding (aOR: 4.73) and haematocrit rise with rapid platelet drop (aOR: 2.74). Conclusion: A focus on sensitivity, specificity, predictive values and association of a number of particular WS should be emphasized in order to better predict severe dengue outcomes.