Malaria and Its Prevention: Socio-acceptability in the Application of Insecticides-treated Bed Nets Among Household Heads in the Rural Village Community of Mazabuka, Zambia

@#Introduction: Malaria poses immerse public health challenges in the world as it is still causing significant morbidity and mortality especially in endemic regions such as Zambia. One of the effective tools in preventing malaria is the application of insecticide-treated bed nets (ITNs). This study aims to explored the socio-acceptability of malaria and ITNs among rural village community of Mazabuka, Zambia. Methods: A semi-structured questionnaire was administered to 177 respondents via two different approached methods; focus group discussions and one-to-one interviews to assess the level of KAP of the respondents about malaria and ITNs. Results: Most of the respondents (98.7%) possessed ITNs that were provided free of charge by the Ministry of Health Zambia. Overall, the knowledge and attitude of respondents on malaria and ITNs were at moderate levels of 68.1% and 71.8%, respectively. In contrast, the level of practice was poor with only 36.2% of positive response. Although 92.1% of the respondents answered correctly on the association between malaria and mosquito bites, myths and misconceptions were still common as some of them still attributed malaria to drinking dirty waters (32.8%), bad weather (15.8%), witchcraft (3.4%), and bathing dirty water (19.8%). The practice was significantly associated with knowledge (p=0.003), but not attitude (p=0.230). Logistic regression analysis revealed that respondents with high knowledge level and tertiary education were more likely to use ITNs correctly (OR=2.957; OR=21.739, respectively). Conclusion: The present study showed that the knowledge gaps were remained among the villagers as misconceptions and their believe of myths were still exist.

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.

A Prevalence Study of Pharyngitis and Its Associated Factors Among Adults With a Sore Throat in Three Primary Clinics in Selangor, Malaysia

@#Introduction: Since pharyngitis in adults is one of the most common infectious diseases seen in general practitioner consultations in Malaysia, data on pharyngitis among adults concerning to its prevalence, socio-demographic, risk factors and clinical manifestations is very much lacking. This study aims to determine the prevalence of pharyngitis among adults in Sepang, Selangor, Malaysia from 2016 to 2017 and its associated demographic and risk factors. Methods: We conducted a cross-sectional study on 215 adult patients with a sore throat as the main symptom and who did not receive any antibiotic treatment within two weeks at three Malaysian primary care clinics. The researchers assessed the participants’ clinical manifestations and collected throat swabs for culture to determine the presence of group A streptococcus (GAS). Data on demographic characteristics, clinical manifestation and throat swab culture results were analyzed using chi-square test and multivariate logistic regression. Results: Pharyngitis was diagnosed in 130/215 (65%) adults with a sore throat. Only six isolates (2.8%) were identified as GAS. The overall mean age ± S.D was 36.43 ± 15.7. The majority of the participants were in the age group of 18-28 years. There were 42.3% males and 57.7% females; most participants were Malay 62.8%, followed by 30.2% Indian, 5.1% Chinese, and 1.9% other ethnicities. The most common symptom among the participants was cough 196 (91.2%), followed by rhinorrhea 161 (74.8%), tonsillar swelling or exudates 68 (31.6%), inflamed or reddish of pharynx 62 (28.8%), swollen anterior cervical lymph nodes 50 (23.3%), and fever ≥37.5°C 28 (13.0%). Conclusion: Besides, there was no significant association between pharyngitis and the demographic variables; the current findings emphasized that inflamed or reddish pharynx, tonsillar swelling or exudates were among the factors associated with pharyngitis.

In Vitro Assessment of Biofilm Formation by Streptococcus pyogenes Isolates From Invasive and Non-invasive Samples With Diverse emm Type Profiles

@#Introduction: Biofilm is one of the important virulence factors that is responsible for the severity and progression of the Streptococcus pyogenes diseases. M-protein is involved in the irreversible attachment of S. pyogenes to surfaces during biofilm development. This study aims to determine the propensity of S. pyogenes to form biofilms and the molecular epidemiology of S. pyogenes isolates by emm typing. Methods: We screened 45 S. pyogenes isolates for the biofilm formation by Congo red agar (CRA) and quantified the biofilms by crystal violet microtiter-plate methods (CVMtP). The emm typing of all isolates was performed by conventional PCR with established primers according to the CDC protocol. Results: Majorities of S. pyogenes were isolated from non-invasive, 27 (60.0%) than invasive sources, 18 (40.0%). Regardless of invasiveness, 40 (88.9%) S. pyogenes isolates formed black colonies on CRA, while 43 (95.6%) of the isolates demonstrated various degrees of biofilm formation by CVMtP method. A total of 30 different emm types and subtypes were identified. No new emm types/subtypes were detected. The predominant emm types/subtypes were emm1, emm63, emm18.21, emm91, and emm97.4 which each gene accounted for 7.0%. All emm types/subtypes of S. pyogenes produced biofilms by CVMtP method except emm17.2 and emm57 which were isolated from non-invasive sources. Conclusions: Biofilm-producing S. pyogenes strains of various sources are genetically diverse and biofilm phenotypes are inherent to individual characteristic rather than specific emm type. Nonetheless, higher propensity of GAS to form biofilms warrants better management strategies to avoid treatment failures in the future.

Susceptibility Profiles of Bacterial Pathogens Causing Community-Acquired Urinary Tract Infections

@#Introduction: Uncontrolled empirical treatment of urinary tract infections (UTIs) has negative aspect on predicting the emergence of antimicrobial resistance and knowledge of those patterns has become extremely important from time to time. Therefore, the aim of the present study was to check the prevalence and resistance patterns of uropathogens in the community acquired UTIs. Methods: A total of 7132 urine samples were combined from male 3131 (43.9%) and female 4001 (56.1%) outpatients suspected of having UTIs, respectively over a three-year period and cultured on routine culture media. The bacteria have been identified using basic biochemical tests, and sensitivity to various antibiotics was determined by the method of disk diffusion. Results: Of 7132 urine samples 797 (11.2%) yielded significant uropathogens. Among the bacterial species, Escherichia coli was the major causative agent of UTIs for both gender (63.7%), followed by Klebsiella spp (20.8%), Enterococcus faecalis (5.3%), Pseudomonas spp (4.1%), Proteus spp (3.1%), Enterobacter spp (1.5%), Candida albicans (0.6%), Staphylococcus saprophyticus (0.5%), Providencia spp (0.1%) and Staphylococcus aureus (0.1%). The antibacterial sensitivity testing for E. coli, to commonly used antibiotics were showed variable resistant as follows: Ampicilln (78%), Amoxicillin (71%), trimethoprim sulfamethoxazole (42%), Amox/clav. (14%) gentamicin (20%), nitrofurantoin (11%), nalidixic acid (22%), ciprofloxacin (20%), Imipenem (16%),Ceftazidim (26%),Cefotaxim (25%),Ceftriaxon (21%),Cefuroxim (33%). Conclusions: The findings showed that antimicrobial resistance patterns of uropathogens in variable, and continuous monitoring of resistance patterns by using of antibiotic susceptibility testing in the laboratory is the most appropriate to treat UTIs rather than the choice of UTIs empirical treatment.

A Case of Neurocysticercosis in Immigrant Worker Presented with Seizures

@#Neurocysticercosis is an infection of the central nervous system (CNS) by larval stage of Taenia solium (pork tapeworms) called cysticerci. Humans may acquire the infection through ingestion of T. solium embryonated eggs after consuming contaminated undercooked food or water. Being a multiracial country where pig farming is limited and consumption of raw or undercooked food is an uncommon practice, the prevalence of neurocysticercosis in Malaysia is presumably low. However, through immigration and international travel, cases of neurocysticercosis may go underreported. Due to unfamiliarity with the disease, cases of neurocysticercosis also may be missed in Malaysia. Therefore, thorough history taking and physical examinations with high index of suspicion are required by the clinicians to warrant proper investigations in order to meet the diagnosis of neurocysticercosis. Patients diagnosed with neurocysticercosis should be treated cautiously with anthelmintic, to avoid unwanted overwhelming immunological response that can lead to unfavourable outcomes. This is a case report of a foreign worker who presented with first onset of focal seizure associated with intermittent headache.

Fibronectin-binding Protein F1 (prtF1) Gene Is Highly Distributed in Tetracycline-resistant, Erythromycin-sensitive Streptococcus pyogenes Isolates

@#Introduction: The internalization process of group A streptococci (GAS) into human cells is one of the crucial steps in the pathogenesis of GAS infections, which could also affect their susceptibility responses toward several antibiotics. Currently, data on the distribution of internalization-associated genes and susceptibility patterns are still lacking in Malaysia. This study investigated the distribution of fibronectin-binding protein F1 (prtF1) and streptococcal pyrogenic exotoxin B (speB) genes in GAS isolates with their susceptibility profiles and source of samples. Methods: We used 43 GAS isolates from our previous stock culture and performed antibiotic susceptibility testing by Kirby-Bauer disk diffusion method and interpreted the results according to the established guidelines. We detected virulence (prtF1 and speB) and resistance (ermA, ermB, mefA, tetM and lnuA) genes by PCR method using established primers and protocols. Results: High resistance rates were observed against doxycycline (58.1%) and clindamycin (16.3%). In comparison, 100.0% and 46.5% of GAS isolates carried speB and prtF1 genes, respectively. tetM and lnuA genes were detected in all respective resistant isolates (100% for each). No macrolide resistance genes were detected. Interestingly, prtF1 gene was highly distributed in doxycycline-resistant than doxycycline-sensitive isolates (60.0% versus 27.8%). Conclusions: High resistance rate of GAS toward doxycycline in our study may potentially reflect the uncontrol dissemination of tetM gene among our isolates. The presence of prtF1 gene among this strain would enhance its ability to evade the intracellular action of antibiotics, which may affect the management of GAS diseases. Thus, close monitoring of GAS by molecular methods is required in the future.

Plasmodium ovale Malaria and Dengue Co-infection in a Glucose-6-Phosphate Dehydrogenase (G6PD) Deficiency Patient : A Case Study

@#Malaria and dengue are among the most important public health threats in Malaysia. These two-arthropod borne diseases have overlapping mosquito biotopes and clinical manifestations, and co-infections have been associated with increased severity notably on the haematological abnormalities. Dengue caused by four dengue virus (DENV) serotypes has been highly endemic in Malaysia. However, malaria due to Plasmodium ovale (P. ovale) has been rarely reported among Malaysian population. Nonetheless, climate change and increased influx of international travellers and migrants have shifted the parasite boundaries to non-endemic countries. Thus, diagnosis and management of imported malarial infections should rely on the geographical knowledge on the origin of potential Plasmodium species, prompt laboratory testing and public health intervention. Moreover, it would be difficult to clinically differentiate dengue fever (DF) with a potential relapse or partially treated case of P. ovale, and there is absolutely no transmission of this Plasmodium species in our country. Hence, we believed that this case deserved to be reported.

Exposure to Microbial Contaminants in Metalworking Fluids (MWF) and the Fractional Exhaled Nitric Oxide (FeNO) Levels among Machining Industry Workers

Abstract@#Introduction: Water based Metalworking fluids (MWF) are commonly used in machining industries and are excellent media for microorganism growth. The study aimed at determining the relationship between the airway inflammation as indicated by fractional exhaled nitric oxide (FeNO) with the microbial contaminants of MWF in aerosol and bulk sample as well as the workers’ reported respiratory health symptoms. Methods: This cross sectional study was carried out on 138 machining workers. Their FeNO were measured using NIOX-MINO instrumentation. The microbial assessments of bacteria and fungus were carried out on the MWF bulk samples and the aerosol using a sampler DUO SAS SUPER 360TM. Results: Findings showed significant difference in the FeNO levels in workers from various job sections (p=0.01). Significant relationships found between high FeNO levels with their closeness to the machines (p=0.03), high number of machines in the workplaces (p=0.02), high environmental bacteria colonies (p=0.04), longer employment years (p<0.001) and more frequent cough reported (p=0.03). Conclusion: Risk factors in the workplace which contributed to higher airway inflammation include their short distance and high number of machines, high environmental bacteria colonies in aerosol and bulk samples as well as longer employment years. Exposures to MWF had also resulted in significantly increased coughing among the respondents.

Antifungal and antibiofilm activity of Persian shallot (Allium stipitatum Regel.) against clinically significant Candida spp

@#Candida species are the most common cause of fungal infections that range from non-life-threatening mucocutaneous illness to life-threatening invasive processes that may involve virtually any organ. Such a broad range of infections requires an equally broad range of therapeutic approach. Persian shallot (Allium stipitatum Regel.) is a medicinal plant that has been widely used in tradition Persian medicine for various ailments. Allium stipitatum is also used in modern medicine and has been reported to have a range of health benefits including antibiotic (antifungal) properties. The present study assessed the in vitro anticandidal and antibiofilm potential of hexane (ASHE) and dichloromethane (ASDE) extracts of Allium stipitatum (Persian shallot) against planktonic and biofilm forms of 5 medically important Candida spp. Antifungal activity was assessed by disk diffusion, minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC) and time-kill assay. The antibiofilm activity of ASHE and ASDE against reference strain C. albicans ATCC 14053 was determined by XTT [2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide] reduction assay. The zone of inhibition ranged from 22 to 40 mm, while the MICs ranged from 8 to 32 μg mL-1. The MFCs of ASHE and ASDE were in the range of 16 to 32 μg mL-1 each respectively. Time-kill kinetics showed that both extracts were strongly fungicidal against planktonic cultures of C. albicans with ~ 1.45 log reduction in CFU at 4 h post-treatment (hpt). In addition, both ASHE and ASDE were shown to inhibit preformed C. albicans biofilms in a concentration-dependent manner. The results demonstrated that ASHE and ASDE were broad-spectrum in action, and could be developed as a promising alternative to synthetic antifungals in controlling infections due to Candida spp. of clinical significance.

Vancomycin-resistant Enterococcus faecium of multi locus sequence type 18 in Malaysia

Vancomycin-resistant Enterococcus faecium (VREF) in human infections mostly belong to the high-risk, epidemic, clonal complex-17 (CC17) group. Treatment limitation and high conjugation frequency makes it dominant in hospitals worldwide. We investigated positive cultures by Pulse-field gel electrophoresis (PFGE), multi locus sequence typing (MLST). DNA of two strains (A2 and C) appeared to be clonally related by PFGE. Three strains were of ST 18 type (A1, B and C) and strain A2 is of a new ST 596. This ST 18 type strain found in our study is crucial and is believed to be the first in Malaysia.

Characterization and antimicrobial activities of two Streptomyces isolates from soil in the periphery of Universiti Putra Malaysia

This study was to assess the identification and antimicrobial activities of two actinomycete isolates. The two isolates designated as B8 and C2, were isolated from a patch of soil in the peripheral area of Universiti Putra Malaysia by streaking on starch casein agar after standard serial dilution procedures. Their antimicrobial activities were first evaluated against eight clinical laboratory strains namely Bacillus sp., Enterococcus sp., Escherichia coli, Klebsiella sp., Pseudomonas sp., Salmonella sp., Staphylococcus aureus, and Staphylococcus epidermidis by perpendicular streak method on Mueller Hinton and Tryptic Soy agar. In both media, a broad-spectrum antibacterial activity was observed for both isolates, with B8 against all the test bacteria and C2 against five of them (Bacillus sp., E. coli, Pseudomonas sp., S. aureus and S. epidermidis). Re-assessment against E. coli ATCC 25922 and S. aureus ATCC 25923 strains by similar method showed antibacterial activities by isolate B8 against both ATTC strains while C2 only against S. aureus ATCC 25923. Streptomyces griseus ATCC 10137 was included in the later experiment and showed antibacterial activity against both ATCC strains. Subsequently, the two isolates were identified by PCR/sequencing techniques and phylogenetic analysis to be Streptomyces species (>93% homology based on 16S rRNA and rpoB genes). Characterization on cultural characteristic and viable count at different temperatures (37ºC and 28ºC), on different microbiological media (AIA, ISP-2, MHA, NA, PDA and TSA), were performed. More morphological features were observed on ISP-2 for both isolates. A higher growth yield was also observed at 28ºC in all media but in comparing that between the two isolates, isolate B8 outnumbered C2 at all experimental conditions. The observed variation in cultural traits and growth yield indicate unique properties between the two antibiotic-producing isolates

In vitro Antiplasmodial Activity and Cytotoxicity of Ten Plants Used as Traditional Medicine in Malaysia / Jurnal Sains Kesihatan Malaysia

Dichloromethane and methanolic extracts of each plant were tested for their antiplasmodial activity on chloroquineresistant strain of Plasmodium falciparum (FCB strain), based on lactate dehydrogenase activity. Cytotoxicity was assessed with the MTT test on MRC-5 human diploid embryonic lung cells. Most extracts of ten selected plants used in Malay traditional medicine in Malaysia had activity in vitro. This supports continued investigations of traditional medicine in the search for new antimalarial agent. The compounds responsible for the observed antiplasmodial effects are under investigation.