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The aim of this study was to determine the effects of two types of coumarin derivatives, namely, 3-acetylcoumarin (AC) and coumarin thiosemicarbazone (CT) on surface characteristics such as the roughness, hardness, and morphology of resin-modified glass-ionomer cement (RMGIC). The release of coumarin from the fabricated RMGIC was also investigated. AC and CT at 1.0% (w/w) concentration were added into 0.3 g of RMGIC powder and mixed with 0.1 g of polyacrylic acid. The fabricated RMGIC-AC and RMGIC-CT were evaluated for surface characteristics such as roughness, hardness and topography. The coumarin release of AC and CT from RMGIC was also determined. The RMGIC-CT demonstrated a decreased roughness value among the materials. The surface roughness exhibited by the RMGIC was statistically higher (p < 0.05) than that of fabricated materials. The RMGIC-AC was observed to have the highest hardness value compared to the RMGIC-CT and RMGIC, and this value was significantly higher (p < 0.05). The surface morphologies of the RMGIC-AC and RMGIC-CT showed a number of pores and irregular surfaces. Meanwhile, the surface roughness value of the RMGIC was statistically higher (p < 0.05) than both RMGIC-AC and RMGIC-CT. In conclusion, the large-sized AC particles significantly increased the surface hardness of the fabricated RMGIC. Conversely, the particle size of both coumarins were not influenced the surface roughness value and the coumarin release profile of fabricated RMGICs.

Hajj pilgrimage is a nobligation for every Muslim that must be carried out at least once in their lifetime. Every year, more than 3 million pilgrims from over 180 countries travel to worship within a confined area of twelve kilometres in Makkah, Kingdom of Saudi Arabia (KSA). The host country and participating countries need to institute effective measures for a safer and healthier Hajj pilgrimage. In the immediate Hajj season during the COVID-19 pandemic, the number of pilgrims need to be reduced to the optimum number that can be managed individual countries’ Hajj missions. Beside proof COVID-19 clearance before travelling and completed Covid-19 vaccination, the stay in KSA should be shortened for Hajj ritual only. The participating countries responsible to thoroughly select individuals with low risk of severe infections, provide health education program, offer all compulsory and recommended vaccines and allocate adequate healthcare/isolation centres in KSA. The participating countries are also liable to prepare adequate quarantine places for hajj pilgrims when returning home. Individual pilgrims are responsible to prevent respiratory pathogens from spreading. Prior to departure for Hajj, they need to undergo a thorough medical examination, received all compulsory and recommended vaccines and follow health education program. During Hajj, they need to follow instructions by Hajj authorities, practicing proper hygiene, wearing a face mask and avoid crowded places. They also need to report to countries’ Hajj mission healthcare if having any fever or respiratory syndromes. With adequate measures prior, during and post-Hajj, we hope health security risk during the next Hajj seasons will be minimized. Bangladesh Journal of Medical Science Vol.20(4) 2021 p.732-740

The flood disaster in Kelantan in 2014 had resulted in substantial health implications including increased cases of communicable diseases. There was a lack of community preparedness including customized health educations in the prevention and control of flood-related communicable diseases in the affected areas. The research was aimed to evaluate the effectiveness of community-based health education modules on flood-related communicable diseases among communities in Kelantan. Health education modules focusing on major food-related diseases were developed. A non-randomized community-controlled trial using the modules were conducted. Outcomes were assessed on knowledge, attitude and preventive practice scores to flood-related communicable diseases using a pre-validated questionnaire. Independent t test was used to compare mean scores between the intervention community (Tumpat) and the control community (Bachok) at 1-month post intervention. One-way independent ANOVA test was done to compare score differences at baseline (pre), post 1-month and post 2-month from repeated surveys among random samples within the intervention community. There were significant improvements in all knowledge components from 9.4% to 52.6% with 10% increment in attitude scores toward preventing behaviours on flood-related communicable diseases. When compared against the control community at one-month post-intervention, there were significantly higher knowledge on types of diseases, symptoms and risk factors as well as practice scores of drinking safe water and protective habits. This research demonstrated that community-based health education is effective in improving relevant knowledge, attitude and preventive practices among affected communities as part of their preparedness toward communicable diseases related to flood.

Bacterial respiratory tract infections (RTIs) are prone to be associated with serious health problems during the annual Hajj pilgrimage and are a public health concern due to the potential of pathogens transmission across continents. This study aimed to perform a diagnostic screening of intended bacteria associated with RTIs among Malaysian Hajj pilgrims by using a newly developed PCR assay. Expectorated sputum specimens (

This study was aimed to develop and evaluate the efficacy of a health education program for increasing knowledge, changing attitudes, and promoting prevention practices to reduce the incidence of RTIs among Malaysian Hajj pilgrims. A quasi-experimental study was done among 130 Hajj pilgrims attending Hajj orientation course organized by a private Hajj tour companies. Hajj pilgrims assigned to intervention group (n = 65) and control group (n = 65). Data were collected baseline and after return from Hajj using a validated questionnaire. Mixed design repeated measure ANOVA was used to analyse the effect of group, time, and group-time interaction on the dependent variables. There was a significant improvement in knowledge, attitude and practice scores in the intervention group compared to the control group. Based on the findings of the interaction of time and group, there was a significant statistical difference for post intervention for knowledge (p = 0.005), attitude (p = 0.041), and practice (p = 0.002). The newly-established health education intervention smartphone application was effective in improving KAP toward RTIs prevention among pilgrims.

Background: Overcrowding during the annual Hajj pilgrimage has been known to increase the risk of infectious diseases transmission. Despite the high prevalence of respiratory illness among Malaysian Hajj pilgrims, knowledge about the etiologic pathogens is yet very limited. Thus, this study aimed to determine the spectrum of bacterial respiratory pathogens among the Hajj pilgrims returning to Malaysia in year 2016. Methods: Expectorated sputum specimens were collected from the Hajj pilgrims with symptomatic respiratory tract infections (RTIs). Subsequently, the bacterial pathogens were identified using the standard bacteriological culture method and Vitek II system. Results: This study indicated that 255 (87.33%) out of 292 cultured sputa were positive with at least one potential pathogenic bacteria. Out of 345 total bacterial isolates, 60% (n = 207) were Haemophilus influenzae, which was associated with both single bacterium infection (132/173, 76.3%) and multiple bacterial infections (75/82, 91.5%). The other bacterial isolates included; Klebsiella pneumoniae (n = 37, 10.7%), Moraxella catarrhalis (n = 27, 7.8%), Haemophilus parainfluenzae (n = 25, 7.2%), Streptococcus group G (n = 18, 5.2%), Klebsiella spesies (n = 16, 4.6%), Streptococcus pneumoniae (n = 11, 3.2%) and few other organisms. Conclusion: High frequency of H. influenzae was isolated from Malaysian Hajj pilgrims, especially those with respiratory symptoms. Further study should evaluate the actual pathogenicity of the organism and the interactions between the respiratory microbiota towards developing effective prevention strategies of RTIs among the local pilgrims. Keywords: Haemophilus influenzae, Respiratory tract infections, Hajj, Infectious diseases

A thermostabilized, multiplex polymerase chain reaction (mPCR) assay was developed in this study for the detection of six respiratory bacterial pathogens. Specific primers were designed for an internal amplification control (IAC) and six target sequences from Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Mycobacterium tuberculosis, and Haemophilus influenzae. The resultant seven-band positive amplification control (PAC) of this heptaplex PCR assay corresponded to 105 base pairs (bp) of IAC, 202 bp of K. pneumoniae, 293 bp of S. aureus, 349 bp of S. pneumoniae, 444 bp of P. aeruginosa, 505 bp of M. tuberculosis, and 582 bp of H. influenzae. Results found that 6% (w/v) of the stabilizer was optimum to preserve the functional conformation of Taq DNA polymerase enzyme. This assay was stable at ambient temperature for at least 6 months. The sensitivity and specificity of this assay were both 100% when testing on the intended target organisms (n = 119) and non-intended species (n = 57). The mPCR assay developed in this study enabled accurate, rapid, and simple detection of six respiratory bacteria.

Reduced susceptibility in Burkholderia pseudomallei during carbapenem therapy may lead to treatment failure. We isolated a clinical strain that had developed reduced susceptibility to carbapenems while on treatment. After reviewing the patient's clinical notes, the initial isolate (BUPS01/14) was exposed to carbapenem in vitro to mimic the clinical scenario. The stability of susceptibility of the carbapenem-exposed strain (BUPS01/14R) was examined by serial subculture in antibiotic-free broth. Biochemical and morphological comparison was performed by the VITEK® system and electron microscopy. MICs increased 32-fold following carbapenem exposure and became stable in the antibiotic-free environment. On electron microscopic examination, the BUPS01/14R cells were smoother and less wrinkled compared to BUPS01/14 cells. This report highlights a potential anti-melioidosis treatment failure due to the emergence of resistance while on carbapenem monotherapy. Further study of this strain is necessary to understand the mechanism of resistance at a molecular level.

Background This study was aimed at demonstrating the impact of a health education intervention on reducing the incidence of influenza-like illnesses symptoms among Malaysian's Hajj pilgrims. MethodsThis study utilizes a quasi-experimental study in the evaluation of the impact of the intervention. Participants were recruited during Hajj orientation courses organized by private Hajj companies. Participants from two separate companies were recruited into an intervention group and control group respectively. The intervention group received a Health Belief Model (HBM) based smartphone health education intervention application while the control group received a normal Hajj guide intervention smartphone application before departure to Hajj. Data on the incidence of influenza-like illnesses (ILI) were collected from participants from two Hajj companies before and after returning from Hajj. Data analysis was conducted using SPSS with descriptive analysis, and analytical tests were conducted at 5% significance level. ResultsA total of 102 pilgrims completed the study in both intervention and control groups. The incidence of the symptoms of influenza-like illnesses (ILI) was statistically significant between the intervention and control groups (p = 0.049). The change in the level of knowledge and attitude among the intervention was statistically significant (p= 0.001, p = 0.029). ConclusionIn conclusion, the health education intervention has an impact on reducing the incidence of symptoms among Hajj pilgrims.

BackgroundRespiratory tract infections are one of the common infection associated with Hajj pilgrimage that is of great public health and global concern. This study is aimed at determining the factor structure of the knowledge, attitude, and practice questionnaire for the prevention of respiratory tract infections during Hajj by confirmatory factor analysis (CFA).MethodsA multistage cluster sampling method was conducted on Malaysian Umrah pilgrims during the weekly Umrah orientation course. A total of 200 Umrah pilgrims participated in the study. The knowledge, attitude and practice (KAP) questionnaire was distributed to pilgrims at the beginning of the orientation and retrieved immediately at the end of the orientation. Data analysis was done using R version 3.5.0 after data entry into SPSS 24. The robust maximum likelihood was used for the estimation due to the multivariate normality assumption violation. A two-factor model was tested for measurement model validity and construct validity for each of the attitude and practice domains.ResultsCFA of a 25-item in total, the two-factor model yielded adequate goodness-of-fit values. The measurement model also showed good convergent and discriminant validity after model re-specification. A two-factor model was tested for measurement model validity and construct validity for each of the attitude and practice domains. The result also showed a statistically significant value (p 2(df) values of 76.8 (43) and 121 (76) for attitude and practice domains, respectively.ConclusionThe KAP questionnaire was proven to have a valid measurement model and reliable constructs. It was deemed suitable for use to measure the KAP of Hajj and Umrah pilgrims towards the prevention for all respiratory tract infections.

Introduction: Influenza-like illnesses (ILI) were defined as a triad of cough, sore throat and fever. The aim of this study is to determine the changes in knowledge, perceptions and practices towards preventive measures of succumbing to ILI before and after hajj among Malaysian hajj pilgrims. Methodology: An openlabeled randomized control trial study performed on five hundred Malaysian hajj pilgrims in the year 2013. After validated pre-hajj questionnaires completed, 250 subjects were given health education, benzalkonium -based handrub and demonstration of the handrub usage together with pamphlets and labelled as an intervention group. Another 250 subjects from another flight batch were given the same questionnaires without health education with unmedicated hand lotion as a control group. The knowledge, perception, practices and compliances to the handrub practice in both groups were compared with the pre-hajj results. Results: No significant difference between pre and post hajj knowledge and practice among hajj pilgrims in intervention group. Pre hajj perception score [ M (IQR) = 30 (5.00), Z= -2.441, p < 0.05 ] is much higher compared to post hajj perception [ M (IQR) = 29 (6.00), Z = -2.441, p < O.O5 ]. No significant difference between pre and post hajj knowledge, perception and practices among hajj pilgrims in control group. Hand hygiene compliance was not improved with provision of health education and free handrub supply. Conclusion: No significant changes in knowledge, perceptions and practices of Malaysian hajj pilgrims pre and post-hajj in both groups. Provision of handrub did not improved hajj pilgrims compliance.

The prevalence of respiratory illness has continued to surge among Hajj pilgrims from different countries despite having some practices of preventive measures. Respiratory illnesses during Hajj could be due to many reasons and many factors that promote disease spread. These factors include overcrowding, cigarette smoking, and direct contact with infectious agents particularly viruses promote the spread of respiratory infections. However, due to the longer duration of the pilgrimage, there are high chances of pilgrims contracting various respiratory illnesses due to exposure to respiratory pathogens. Hajj pilgrims' knowledge, attitudes, and practices toward respiratory tract infections are used as the determinant of the effectiveness of the health education interventions. Knowledge and application of basic hygiene principles, use of face masks, following cough etiquettes, engaging in social distancing, and engaging in other measures are highly important. In this paper, we reviewed the various effective intervention strategies implemented to help prevent respiratory tract infections during Hajj.

Background Hajj pilgrimage faces numerous challenges including a high prevalence of respiratory tract infection as well as its prevention strategies. The aim of this study was to develop and validate a questionnaire to evaluate knowledge, attitude and practice (KAP) towards respiratory tract infections (RTIs) prevention among Malaysian Hajj pilgrims. Methods This study was conducted among Malaysian Umrah pilgrims in Malaysia from Kuala Lumpur and Kelantan. The questionnaire then underwent a series of validation process that included content, face validity and exploratory part. Item response theory (IRT) analysis was utilized for the validation of the knowledge domain. The attitude and practice were validated using the exploratory factor analysis (EFA). Results The validation process resulted in a questionnaire that comprised of four main sections: demography, knowledge, attitude, and practice. Following IRT analysis of the knowledge domain, all items analyzed were within the acceptable range of difficulty and discrimination. The Kaiser-Meyer-Olkin measure of sampling adequacy (KMO) was 0.72 and 0.84 for attitude and practice domain respectively and Bartlett’s test of Sphericity for both domains were highly significant (P 0.3). The Cronbach’s alpha for reliability of the knowledge, attitude and practice domains all showed acceptable values of > 0.6 (0.92, 0.77 and 0.85). Conclusion The findings of this validation and reliability study showed that the developed questionnaire had a satisfactory psychometric property for measuring KAP of Malaysian Hajj pilgrims.

Respiratory tract infection (RTI) is a major public health challenge during the Muslim pilgrimage to Makkah. This study aims to evaluate the knowledge, attitude, and practice of Malaysian Hajj and Umrah pilgrims towards the prevention of RTIs in 2018 and determine correlations among three domains. A cross-sectional study was conducted among 225 Umrah and Hajj pilgrims. Knowledge, attitude, and practice (KAP) towards RTI prevention was assessed by using a validated self-administered questionnaire among pilgrims attending a weekly orientation course organized by private Hajj/Umrah companies. Out of 225 participants, 65.9% of respondents were female with the mean (SD) age of 46.74 (13.38) years. The interquartile range (IQR) score for knowledge is 18.0 (6.0), the mean scores of attitude and practice are 32.65 (4.72) and 25.30 (4.9). respectively. Significant and negative linear correlations between knowledge and practice (r = &minus, 0.232, p &lt, 0.001), and attitude and practice (r = 0.134, p = 0.045) were observed. Results from the current study showed good knowledge of RTIs among Malaysian pilgrims. However, a poor attitude was reflected in their preventive practice behaviors. This will further help in the prevention and management of RTIs during Hajj and Umrah. Therefore, an extensive educational health campaign should be provided to pilgrims to create awareness.

Although doxycycline is active against Burkholderia pseudomallei and has been used in the eradication stage of melioidosis therapy, it is not regularly used during the initial intensive phase. In order to assess its potential use in intensive phase therapy, we investigated in vitro pharmacodynamic activity of doxycycline and β-lactams alone and in combination against four Malaysian strains of B. pseudomallei. Using a checkerboard assay, the combinations of doxycycline and imipenem, doxycycline and ceftazidime, and doxycycline and amoxicillin–clavulanate tested against four strains showed indifferent effects with summation fractional inhibitory concentration values ranging from 0.62 to 2.12. Time-kill experiments also indicated that the combinations of doxycycline/β-lactam antibiotics against four tested strains did not fulfil synergy criteria, in which all combinations showed indifferent effects with − 1.36 to 1.26-log CFU/mL compared to the most active monotherapy regimen in each combination. No re-growth of bacteria was detected after the early killing in doxycycline/β-lactam combination regimens compared to β-lactam monotherapy regimens, in which 9 out of 10 were associated with re-growth of bacteria. As no synergistic activity was observed, this in vitro study showed that doxycycline offers no additional benefit to be used in combination with β-lactams in the intensive phase of therapy.

This study aimed to assess the uptake of recommended vaccines and to identify the factors associated with the vaccines' uptake among Malaysian Hajj and Umrah pilgrims. A cross-sectional survey among Malaysian Hajj and Umrah pilgrims in 2018. The uptake of the recommended vaccines was surveyed through an anonymous self-administered questionnaire to pilgrims attending a pre-departure Hajj/Umrah orientation course. Descriptive statistics were used for elaborating the demographic characteristics and vaccines uptake of the respondents. Multiple logistic regression was used for predicting the factors associated with the vaccines' uptake. A total of 1,274 pilgrims participated in the study with a mean age (standard deviation) of 42.42 (15.6). A total of 833 (65.4%) participants were females and 232 of the participants (18.2%) had at least more than one chronic disease. The uptake of influenza and pneumococcal vaccines were 28.6% (364/1,274) and 25.4% (324/1,274), respectively. Among the 527 pilgrims who were “at increased risk” of infections, 168 (31.9%) and 184 (34.9%) received influenza and pneumococcal vaccines, respectively. Gender, marital status and occupation were the common predictors associated with vaccines uptake. The vaccination uptake among Malaysian Hajj and Umrah pilgrims is low and declining from previous years. Educating the pilgrims toward vaccine uptake is essential and exploring the barriers for vaccination.

Serratia marcescens previously was thought to be a low pathogenic bacterium but has recently being recognized as a nosocomial pathogen. In May 2007, patients with nosocomial septicemia caused by S. marcescens were identified in a neonatal intensive care unit (NICU) at a tertiary-teaching hospital. The objective of this report is to highlight the rare source of the S. marcescens outbreak. Here, we describe the investigations and outcomes of this outbreak. Case subject was defined as any neonate receiving medical care at the NICU, from one week before the index patient became symptomatic and onwards, with clinical signs and symptoms of sepsis, with or without a positive blood culture, which yielded S. marcescens. Chart reviews were conducted for case subjects. Various samples from the NICU were sent for sterility testing including medications, solutions, used and freshly prepared total parenteral nutrition (TPN) from the NICU and the pharmacy unit. The specimens were processed and positive isolates were identified as S. marcescens using standard microbiological methods. S. marcescens isolates were subjected to pulsed field gel electrophoresis for molecular typing and source identification. S. marcescens were isolated from five neonates, used TPN and freshly prepared TPN of different batches in pharmacy. None of the environmental samples and medications grew S. marcescens. Analysis of macrorestriction patterns of genomic DNA by pulsed field gel electrophoresis demonstrated indistinguishable patterns. This outbreak was related to bacterial contamination of TPN. This report points at a rare source of S. marcescens outbreak. However, targeted investigations and immediate active interventions enabled the managing team to control the outbreak in the NICU.

Antibacterial activity of zinc oxide nanoparticles (ZnO-NPs) has received significant interest worldwide particularly by the implementation of nanotechnology to synthesize particles in the nanometer region. Many microorganisms exist in the range from hundreds of nanometers to tens of micrometers. ZnO-NPs exhibit attractive antibacterial properties due to increased specific surface area as the reduced particle size leading to enhanced particle surface reactivity. ZnO is a bio-safe material that possesses photo-oxidizing and photocatalysis impacts on chemical and biological species. This review covered ZnO-NPs antibacterial activity including testing methods, impact of UV illumination, ZnO particle properties (size, concentration, morphology, and defects), particle surface modification, and minimum inhibitory concentration. Particular emphasize was given to bactericidal and bacteriostatic mechanisms with focus on generation of reactive oxygen species (ROS) including hydrogen peroxide (H2O2), OH− (hydroxyl radicals), and O2 −2 (peroxide). ROS has been a major factor for several mechanisms including cell wall damage due to ZnO-localized interaction, enhanced membrane permeability, internalization of NPs due to loss of proton motive force and uptake of toxic dissolved zinc ions. These have led to mitochondria weakness, intracellular outflow, and release in gene expression of oxidative stress which caused eventual cell growth inhibition and cell death. In some cases, enhanced antibacterial activity can be attributed to surface defects on ZnO abrasive surface texture. One functional application of the ZnO antibacterial bioactivity was discussed in food packaging industry where ZnO-NPs are used as an antibacterial agent toward foodborne diseases. Proper incorporation of ZnO-NPs into packaging materials can cause interaction with foodborne pathogens, thereby releasing NPs onto food surface where they come in contact with bad bacteria and cause the bacterial death and/or inhibition.

The structural morphology and in vitro toxicity of nano- and micro-sized ZnO structures were investigated. All the ZnO samples were characterized to determine their morphologies, particle sizes, structures and optical bandgaps. Transmission electron microscopy and field-emission scanning electron microscopy results revealed that the morphologies of nano-sized ZnO-N1 and ZnO-N2 samples consisted of spherical and irregularly-shaped particles, respectively. The corresponding particle sizes were 20–40 nm and 50–80 nm, respectively. The morphologies of micro-sized ZnO-M1 and ZnO-M2 samples were found to be mostly rod-structures and plate-structures, respectively. Rod diameters were 40–100 nm whereas plate widths were 50–150 nm. ZnO-N1 had the highest toxicity towards the cells, causing the cell viability to be less than 70% for all concentration ranges. This phenomenon was due to the small particle size and release of zinc ions. Micro-sized ZnO-M1 and ZnO-M2 samples had toxicity limits of 0.3 mM at most.

To describe the clinical characteristics and the risk factors associated with mortality in patients with meningitis. This is a retrospective review of patients diagnosed to have meningitis with positive culture of the cerebrospinal fluid (CSF) specimen. All cases aged 19 > years who were admitted to Hospital USM between January 2004 and December 2011 were included in the study. The CSF results database were obtained from the Department of Medical Microbiology and Parasitology, Hospital USM, Kelantan. A checklist was used to record the clinical characteristics. A total of 125 cases met the inclusion criteria. The age of patients ranged between newborn and 19 years old (Mean±SD, 74.5±80.6 months). The majority of them were males (65.6%). Fever was the most common presentation (73.6%) followed by poor oral intake (48.0%), seizure (36.0%) and headache (24.8%). The mortality rate was 31.2%. Coagulase negative staphylococcus was the most frequent pathogens isolated (21.6%), followed by Acinetobacter spp. (17.6%), Staphylococcus aureus (13.6%), Streptococcus spp. (11.2%) and Klebsiella pneumoniae (6.4%). There were significant association of in-hospital death with age (p=0.020) and conscious level (p=0.001). Infectious meningitis is a big health concern, especially among children. We found that coagulase negative staphylococcus, Acinetobacter species, S. aureus, Streptococcus spp and K. pneumoniae were prevalent in our hospital. These microorganisms were hospital associated pathogens. The 31% mortality linked to hospital acquired meningitis specifies the need for focused physician attention especially among younger aged patients.

Background/Purpose Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen responsible for significant numbers of nosocomial and community-acquired infections worldwide. Molecular diagnosis for MRSA nasal carriers is increasingly important for rapid detection and screening of MRSA colonization because the conventional methods are time consuming and labor intensive. However, conventional polymerase chain reaction (PCR) tests still require cold-chain storage as well as trained personnel, which makes them unsuitable for rapid high-throughput analysis. The aim of this study was to develop a thermostabilized PCR assay for MRSA in a ready-to-use form that requires no cold chain. Methods The thermostabilized PCR assay detects the following targets simultaneously: (1) 16S rRNA of the Staphylococcus genus; (2) fem A gene specific for S. aureus ; (3) mec A gene conferring methicillin resistance; and (4) luk S gene, which encodes the virulent toxin. The thermostabilized PCR incorporates an internal amplification control that helps to verify the presence of PCR inhibitors in samples. PCR reagents and specific primers were lyophilized into a pellet form with an enzyme stabilizer. Results The PCR was validated with 235 nasal swabs specimens and was found to be 100% sensitive and specific. The stability of the thermostabilized PCR was evaluated using the Q 10 method and it was found to be stable for approximately 6 months at 24°C. The limit of detection of thermostabilized PCR assay was determined by probit regression (95% confidence interval) was 10 6 colony forming units at the bacterial cell level and 10 ng of DNA at the genomic DNA level, which is comparable with conventional PCR methods. Conclusion A rapid thermostabilized PCR assay that requires minimal pipetting steps and is cold chain-free was developed for detecting MRSA nasal carriers.

Omar Salad Elmi, Ph.D., Mat Zuki Bin Mat Jeab, M.Med., Sarimah Binti Abdullah, M.Med., Habsah Hasan, M.Med., Bin Alwi Zilafil, Ph.D., and Nyi Nyi Naing, M.Med., Unit of Biostatistics and Research Methodology, School of Medical Sciences, Universiti Sains Malaysia, Respiratory Unit, Department of Medicine, Hospital Raja Perempuan Zainab II Kota Bharu, Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Department of Pediatrics, School of Medical Sciences, Universiti Sains Malaysia, Kelantan, Malaysia C A s e R e p o R T

Although doxycycline is active against Burkholderia pseudomallei and has been used in the eradication stage of melioidosis therapy, it is not regularly used during the initial intensive phase. In order to assess its potential use in intensive phase therapy, we investigated in vitro pharmacodynamic activity of doxycycline and β-lactams alone and in combination against four Malaysian strains of B. pseudomallei. Using a checkerboard assay, the combinations of doxycycline and imipenem, doxycycline and ceftazidime, and doxycycline and amoxicillin–clavulanate tested against four strains showed indifferent effects with summation fractional inhibitory concentration values ranging from 0.62 to 2.12. Time-kill experiments also indicated that the combinations of doxycycline/β-lactam antibiotics against four tested strains did not fulfil synergy criteria, in which all combinations showed indifferent effects with − 1.36 to 1.26-log CFU/mL compared to the most active monotherapy regimen in each combination. No re-growth of bacteria was detected after the early killing in doxycycline/β-lactam combination regimens compared to β-lactam monotherapy regimens, in which 9 out of 10 were associated with re-growth of bacteria. As no synergistic activity was observed, this in vitro study showed that doxycycline offers no additional benefit to be used in combination with β-lactams in the intensive phase of therapy.

Klebsiella pneumoniaeandHaemophilus influenzaeare two common pathogens associated with respiratory tract infections. The identification of these pathogens using conventional molecular diagnostic tests requires trained personnel, cold-chain transportation, and storage-dependance, which does not render them user-friendly. The aim of this study was to develop a thermostabilized, cold-chain-free, one-step multiplex PCR for simultaneous detection ofK. pneumoniaeandH. influenzae.The multiplex PCR assay was designed to amplify thephpgene ofK. pneumoniae(202 bp) andp6gene ofH. influenzae(582 bp). In addition, the specific primer to amplifyglmgene ofHelicobacter pylori(105 bp) was included as an internal amplification control. Subsequently, the designed primers and all PCR reagents were thermostabilized by lyophilization. The stability of the thermostabilized PCR was evaluated using theQ10method. The sensitivity and specificity of performances for thermostabilized PCR were evaluated using 127 clinical isolates and were found to be 100% sensitive and specific. The thermostabilized PCR mix was found to be stable for 30 days and theQ10 accelerated stability was found to be 3.02 months. A cold-chain-free, PCR assay for easy, rapid, and simultaneous detection ofK. pneumoniaeandH. influenzaewas successfully developed in this study.

Ultraviolet-A (UVA) radiation is present in sunlight and have been associated with various types of human skin cancers. In this study, rubber-grade zinc oxide (ZnO) powder was used as the targeted materials to study its UVA photoresponse as well as its antibacterial function. ZnO powder was synthesized using French process. The morphological structures of the samples were investigated using field emission scanning electron microscopy and transmission electron microscopy. The dominant morphology of the sample was micro/nanoplate. The optical bandgap of the ZnO sample is 3.19 eV based on the UV-Visible measurement. Current-voltage measurement was conducted to study the effect of UVA (390 nm) illumination on the photoconductivity of the ZnO pellet. Photoconductivity was observed to increase significantly under UVA exposure due to light absorption on the surface of ZnO to raise the electrons across the bandgap. The current response of the UVA-induced also revealed the small persistent photoconductivity after the UVA light was turned off. Besides, higher voltage bias would lead to higher current flow under the same intensity of UVA exposure. According to the antibacterial test towards Staphylococcus aureus, the percentage inhibition of the bacterial after 24 h incubation increase when the concentration of ZnO suspension increases. The UVA illumination had improved the inhibition of the bacterial growth. This is due to the excitation of ZnO and increasing of free charge carriers in the solution, leading to potential distortion to the membrane surface of the bacteria. ZnO powder performed high absorption of UVA and they are not only can be used to block the UVA sunlight, but also have higher antibacterial capability under UVA excitation.

The antibacterial responses of zinc oxide (ZnO) structures against Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus pyogenes were investigated. Two ZnO powder samples, one with rod-like (ZnO-1) and the other with plate-like (ZnO-2) structures, were characterized for their morphological, structural, and optical properties. The rods were 30–120 nm in diameter, and the plates were 40–100 nm thick. XRD results revealed the wurtzite crystallinity of ZnO with average crystallite sizes of 33.72 (ZnO-1) and 39.25 (ZnO-2) nm. ZnO-2 possessed a relatively higher green photoluminescence than that of ZnO-1, suggesting a relatively higher amount of oxygen vacancies in ZnO-2 structures. Optical density measurements showed that both ZnO samples inhibited the growth of S. aureus, P. aeruginosa, and S. pyogenes by 29–98% after 24 h of treatment. The most dramatic growth inhibition was observed in S. pyogenes with 96% and 98% inhibition for ZnO-1 and ZnO-2, respectively, leading to a probable bactericidal phenomenon. The toxicological effect on S. pyogenes was probably due to the absence of catalase, making the bacteria vulnerable to the harmful reactive oxygen species (ROS) released by ZnO. ZnO-1 induced higher inhibition toward S. aureus and P. aeruginosa than that of ZnO-2 because of the smaller particle size of rod structures compared to plate and slab structures. The adhesion of ZnO particles on the membrane of bacteria could be the underlying cause of zinc toxicity effect towards the bacteria. ZnO-1 possessed larger surface area and provided higher amount of zinc atom, thereby inducing higher level of toxicity toward the bacteria. Two possible mechanisms were proposed to explain the inhibition of bacteria, namely, ROS toxicity toward cellular constituents and interaction of zinc with bacteria membrane through adhesion of ZnO particle. Several ZnO morphological-antibacterial correlations were presented in this work.

The effects of surface modification of zinc oxide (ZnO) powder and UVA illumination on the powder towards Escherichia coli and Staphylococcus aureus were investigated. FESEM-EDS results showed that oxygen annealing increased the O:Zn ratio on the surface of ZnO-rod and ZnO-plate samples. Surface conductances of ZnO-rod and ZnO-plate pellets were reduced from 1.05 nS to 0.15 nS and 1.34 nS to 0.23 nS, respectively. Meanwhile, UVA illumination on the surface of the ZnO-rod and ZnO-plate samples was found to improve surface conductance to 7.08 nS and 6.51 nS, respectively, due to the release of charge carrier. Photoluminescence results revealed that oxygen annealing halved the UV emission intensity and green emission intensity, presumably caused by oxygen absorption in the ZnO lattice. The antibacterial results showed that oxygen-treated ZnO exhibited slightly higher growth inhibition on E. coli and S. aureus compared with unannealed ZnO. UVA illumination on ZnO causes the greatest inhibition toward E. coli and S. aureus. Under the UVA excitation, the inhibition of E. coli increased by 18% (ZnO-rod) and 13% (ZnO-plate) while the inhibition of S. aureus increased by 22% (ZnO-rod) and 21% (ZnO-plate). Release of reactive oxygen species were proposed in antibacterial mechanisms, which were aided by surface modification and UVA photoactivation. The reactive oxygen species disrupted the DNA and protein synthesis of the bacterial cell, causing bacteriostatic effects toward E. coli and S. aureus.

ZnO with two different morphologies were used to study the inhibition of Streptococcus sobrinus and Streptococcus mutans which are closely associated with tooth cavity. Rod-like shaped ZnO-A and plate-like shaped ZnO-B were produced using a zinc boiling furnace. The nanopowders were characterized using energy filtered transmission electron microscopy (EFTEM), X-ray diffraction (XRD), photoluminescence (PL) spectroscopy, Raman spectroscopy and dynamic light scattering (DLS) to confirm the properties of the ZnO polycrystalline wurtzite structures. XRD results show that the calculated crystallite sizes of ZnO-A and ZnO-B were 36.6 and 39.4nm, respectively, whereas DLS revealed particle size distributions of 21.82nm (ZnO-A) and 52.21nm (ZnO-B). PL spectra showed ion vacancy defects related to green and red luminescence for both ZnO particles. These defects evolved during the generation of reactive oxygen species which contributed to the antibacterial activity. Antibacterial activity was investigated using microdilution technique towards S. sobrinus and S. mutans at different nanopowder concentrations. Results showed that ZnO-A exhibited higher inhibition on both bacteria compared with ZnO-B. Moreover, S. mutans was more sensitive compared with S. sobrinus because of its higher inhibition rate.

Introduction: Universal hepatitis B vaccination to all newborns have been implemented in Malaysia, since 1989, with nationwide coverage of 96.26%. In this study, we aimed to assess the prevalence of anti-HBs and HBsAg among selected undergraduate students who presumably had completed hepatitis B vaccination during their early childhoods. Methods: Results of hepatitis B screening were obtained retrospectively from 239 newly enrolled undergraduate students at Health Campus, UniversitiSains Malaysia, in 2018. Serum samples were previously tested for the presence of anti-HBs and HBsAg using chemiluminescence immunoassay. Epidemiological data, anti-HBs and HBsAg were analysed using a descriptive statistical method. Results: Anti-HBs was undetected in 82.2 % ( n = 194/236) of the tested students. None of these students had detectable HBsAg. Conclusion: This study showed anti-HBs level weaned after almost two decades of vaccination. Therefore, an alternative new testing algorithm such as the administration booster dose of hepatitis B vaccination can be considered in order to ensure that the new students are protected against potential exposure to hepatitis B during their clinical practices. International Journal of Human and Health Sciences Vol. 03 No. 04 October’19 Page : 245-248

Staphylococcus aureus nasal carriage is a common source of nosocomial infection and colonization. The aim of the present study was to assess the burden of methicillin-resistant S. aureus nasal carriage, its association with factors of interest including its genetic relationships. The prevalence of S. aureus nasal carriage was found to be 28.7%. This study showed that patients with a history of previous antibiotic intake, nasogastric tube, and longer hospitalization had a significantly high risk of being MRSA nasal carriers. The genetic relationship of all 34 nasal MRSA isolates revealed four major clusters of isolates, and there was a relationship between MRSA isolated from inpatients and healthcare workers.