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Optimized conditions for phytoremediation of diesel by Scirpus grossus in horizontal subsurface flow constructed wetlands (HSFCWs) using response surface methodology

Al-Baldawi IA, Sheikh Abdullah SR, Abu Hasan H, Suja F, Anuar N, Mushrifah I

J Environ Manage, 2014 Jul 1;140:152-9.
PMID: 24762527 DOI: 10.1016/j.jenvman.2014.03.007

This study investigated the optimum conditions for total petroleum hydrocarbon (TPH) removal from diesel-contaminated water using phytoremediation treatment with Scirpus grossus. In addition, TPH removal from sand was adopted as a second response. The optimum conditions for maximum TPH removal were determined through a Box-Behnken Design. Three operational variables, i.e. diesel concentration (0.1, 0.175, 0.25% Vdiesel/Vwater), aeration rate (0, 1 and 2 L/min) and retention time (14, 43 and 72 days), were investigated by setting TPH removal and diesel concentration as the maximum, retention time within the given range, and aeration rate as the minimum. The optimum conditions were found to be a diesel concentration of 0.25% (Vdiesel/Vwater), a retention time of 63 days and no aeration with an estimated maximum TPH removal from water and sand of 76.3 and 56.5%, respectively. From a validation test of the optimum conditions, it was found that the maximum TPH removal from contaminated water and sand was 72.5 and 59%, respectively, which was a 5 and 4.4% deviation from the values given by the Box-Behnken Design, providing evidence that S. grossus is a Malaysian native plant that can be used to remediate wastewater containing hydrocarbons.
Fulltext Rapid detection of methicillin-resistant Staphylococcus aureus by a newly developed dry reagent-based polymerase chain reaction assay

Al-Talib H, Yean CY, Al-Khateeb A, Hasan H, Ravichandran M

J Microbiol Immunol Infect, 2014 Dec;47(6):484-90.
PMID: 23927820 DOI: 10.1016/j.jmii.2013.06.004

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.
Fulltext Methicillin-resistant Staphylococcus aureus nasal carriage among patients and healthcare workers in a hospital in Kelantan, Malaysia

Al-Talib H, Yean CY, Hasan H, Nik Zuraina NM, Ravichandran M

Pol J Microbiol, 2013;62(1):109-12.
PMID: 23829087

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.
Fulltext The prevalence of acute respiratory symptoms and role of protective measures among Malaysian hajj pilgrims

Deris ZZ, Hasan H, Sulaiman SA, Wahab MS, Naing NN, Othman NH

J Travel Med, 2010 Mar-Apr;17(2):82-8.
PMID: 20412173 DOI: 10.1111/j.1708-8305.2009.00384.x

BACKGROUND: Respiratory symptoms including cough, runny nose, sore throat, and fever are the most common clinical manifestations faced by hajj pilgrims in Mecca. The aim of the study was to determine the prevalence of respiratory symptoms among Malaysian hajj pilgrims and the effect of a few protective measures taken by hajj pilgrims to reduce respiratory symptoms.
METHODS: A cross-sectional study was conducted by distributing survey forms to Malaysian hajj pilgrims at transit center before flying back to Malaysia. The recruitment of respondents to the survey was on a voluntary basis.
RESULTS: A total of 387 survey forms were available for analysis. The mean age was 50.4 +/- 11.0 years. The common respiratory symptoms among Malaysian hajj pilgrims were: cough 91.5%, runny nose 79.3%, fever 59.2%, and sore throat 57.1%. The prevalence of hajj pilgrims with triad of cough, subjective fever, and sore throat were 40.1%. The symptoms lasted less than 2 weeks in the majority of cases. Only 3.6% did not suffer from any of these symptoms. Seventy-two percent of hajj pilgrims received influenza vaccination before departure and 72.9% wore facemasks. Influenza vaccination was not associated with any of respiratory symptoms but it was significantly associated with longer duration of sore throat. Wearing masks was significantly associated with sore throat and longer duration of sore throat and fever.
CONCLUSIONS: The prevalence of respiratory symptoms was high among Malaysian hajj pilgrims and the current protective measures seemed inadequate to reduce it. Beside standardization of the term used in hajj studies, more collaborative effort should be taken to reduce respiratory symptoms. The hajj authority should prepare for the challenge of pandemic influenza by providing more healthcare facilities and implementation of more strict measures to reduce the transmission of pandemic influenza strain among hajj pilgrims.
Comparative evaluation of five culture media with triplex PCR assay for detection of methicillin-resistant Staphylococcus aureus

Al-Talib H, Yean CY, Al-khateeb A, Singh KK, Hasan H, Al-Jashamy K, et al.

Curr Microbiol, 2010 Jul;61(1):1-6.
PMID: 20033170 DOI: 10.1007/s00284-009-9567-8

The emergence of methicillin-resistant Staphylococcus aureus (MRSA) is responsible for nosocomial and community-acquired infections. Hence, rapid and accurate laboratory diagnosis of MRSA is a vital constituent of control measures. The present study evaluated five different methods for the identification of MRSA. A total of 207 S. aureus clinical isolates that consisted of 89 MRSA and 118 methicillin-susceptible S. aureus (MSSA) strains confirmed by PCR were tested. MRSA strains were evaluated by five different methods: chromogenic MRSA agar (CMRSA), oxacillin resistance screening agar base (ORSAB), mannitol salt oxacillin agar (MSO), mannitol salt cefoxitin agar with two different concentrations of cefoxitin [4 microg/ml (MSC-4) and 6 microg/ml (MSC-6)]. The results of the different methods were compared to mecA PCR as the gold standard. MSC-6 showed only six false-positive MRSA in comparison with PCR. The sensitivities and specificities of MSC-6, MSC-4, MSO-4, ORSAB, and CMRSA were as follows: 98.9/94.9%, 100/83.1%, 89.9/87.3%, 97.8/96.6%, and 95.5/94.9%, respectively. In comparison with PCR, it was found that both MSC-6 and ORSAB were relatively the least expensive screening tests ($0.70 and $1.00, respectively). In conclusion, all methods were comparable, but MSC-6 was the least expensive medium for MRSA screening.
Fulltext Multidrug-resistant tuberculosis and risk factors associated with its development: a retrospective study

Elmi OS, Hasan H, Abdullah S, Mat Jeab MZ, Bin Alwi Z, Naing NN

J Infect Dev Ctries, 2015 Oct;9(10):1076-85.
PMID: 26517482 DOI: 10.3855/jidc.6162

INTRODUCTION: Multidrug-resistant tuberculosis (MDR-TB) has emerged as a major clinical public health threat and challenges the national TB control program in Malaysia. Data that elaborates on the risk factors associated with the development of MDR-TB is highly limited in this country. This study was aimed to determine the risk factors associated with the development of MDR-TB patients in peninsular Malaysia.
METHODOLOGY: This was a case control study; the data were collected from medical records of all the registered MDR-TB patients at five referral TB hospitals in peninsular Malaysia from January 2010 to April 2014. The 105 cases were all confirmed by a positive sputum culture of Mycobacterium tuberculosis for MDR-TB and extensively drug-resistant (XDR)-TB. As a comparison, a total of 209 non-MDR-TB cases were randomly selected as controls.
RESULTS: A total of 105 MDR-TB and 209 non MDR-TB patients were studied. The risk factors associated with MDR-TB within the multivariate analysis were previous tuberculosis treatment, HIV infection, being an immigrant, and high load of positive for acid-fast bacillus (AFB) smear.
CONCLUSIONS: The findings of this study revealed that patients who had received previous treatment for tuberculosis, were infected with HIV, were immigrants, and had a high burden of positive testing for AFB smear were more likely to have MDR-TB. An enhanced understanding of the risk factors associated with MDR-TB strains is imperative in the development of a national policy for public health interventions.
Comparison of the efficiencies of attached- versus suspended-growth SBR systems in the treatment of recycled paper mill wastewater

Muhamad MH, Sheikh Abdullah SR, Abu Hasan H, Abd Rahim RA

J Environ Manage, 2015 Nov 1;163:115-24.
PMID: 26311084 DOI: 10.1016/j.jenvman.2015.08.012

The complexity of residual toxic organics from biologically treated effluents of pulp and paper mills is a serious concern. To date, it has been difficult to choose the best treatment technique because each of the available options has advantages and drawbacks. In this study, two different treatment techniques using laboratory-scale aerobic sequencing batch reactors (SBRs) were tested with the same real recycled paper mill effluent to evaluate their treatment efficiencies. Two attached-growth SBRs using granular activated carbon (GAC) with and without additional biomass and a suspended-growth SBR were used in the treatment of real recycled paper mill effluent at a chemical oxygen demand (COD) level in the range of 800-1300 mg/L, a fixed hydraulic retention time of 24 h and a COD:N:P ratio of approximately 100:5:1. The efficiency of this biological treatment process was studied over a 300-day period. The six most important wastewater quality parameters, namely, chemical oxygen demand (COD), turbidity, ammonia (expressed as NH3-N), phosphorus (expressed as PO4(3)-P), colour, and suspended solids (SS), were measured to compare the different treatment techniques. It was determined that these processes were able to almost completely and simultaneously eliminate COD (99%) and turbidity (99%); the removals of NH3-N (90-100%), PO4(3)-P (66-78%), colour (63-91%), and SS (97-99%) were also sufficient. The overall performance results confirmed that an attached-growth SBR system using additional biomass on GAC is a promising configuration for wastewater treatment in terms of performance efficiency and process stability under fluctuations of organic load. Hence, this hybrid system is recommended for the treatment of pulp and paper mill effluents.
Burden of bacterial meningitis: a retrospective review on laboratory parameters and factors associated with death in meningitis, kelantan malaysia

Basri R, Zueter AR, Mohamed Z, Alam MK, Norsa'adah B, Hasan SA, et al.

Nagoya J Med Sci, 2015 Feb;77(1-2):59-68.
PMID: 25797971

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.
Fulltext Evaluation of female Aedes aegypti proteome via LC-ESI-MS/MS using two protein extraction methods

Shettima A, Ishak IH, Abdul Rais SH, Abu Hasan H, Othman N

PeerJ, 2021;9:e10863.
PMID: 33717682 DOI: 10.7717/peerj.10863

Background: Proteomic analyses have broadened the horizons of vector control measures by identifying proteins associated with different biological and physiological processes and give further insight into the mosquitoes' biology, mechanism of insecticide resistance and pathogens-mosquitoes interaction. Female Ae. aegypti ingests human blood to acquire the requisite nutrients to make eggs. During blood ingestion, female mosquitoes transmit different pathogens. Therefore, this study aimed to determine the best protein extraction method for mass spectrometry analysis which will allow a better proteome profiling for female mosquitoes.
Methods: In this present study, two protein extractions methods were performed to analyze female Ae. aegyti proteome, via TCA acetone precipitation extraction method and a commercial protein extraction reagent CytoBusterTM. Then, protein identification was performed by LC-ESI-MS/MS and followed by functional protein annotation analysis.
Results: The CytoBusterTM reagent gave the highest protein yield with a mean of 475.90 µg compared to TCA acetone precipitation extraction showed 283.15 µg mean of protein. LC-ESI-MS/MS identified 1,290 and 890 proteins from the CytoBusterTM reagent and TCA acetone precipitation, respectively. When comparing the protein class categories in both methods, there were three additional categories for proteins identified using CytoBusterTM reagent. The proteins were related to scaffold/adaptor protein (PC00226), protein binding activity modulator (PC00095) and intercellular signal molecule (PC00207). In conclusion, the CytoBusterTM protein extraction reagent showed a better performance for the extraction of proteins in term of the protein yield, proteome coverage and extraction speed.
Fatal necrotizing pneumonia caused by Panton-Valentine leukocidin-producing hospital-acquired Staphylococcus aureus: a case report

Al-Talib H, Hasan H, Yean CY, Al-Ashwal SM, Ravichandran M

Jpn J Infect Dis, 2011;64(1):58-60.
PMID: 21266757

Panton-Valentine leukocidin (PVL) is a cytotoxin which causes leukocyte destruction and tissue necrosis. Although it is produced by fewer than 5% of Staphylococcus aureus strains, PVL-producing S. aureus is emerging as a serious problem worldwide. There has been a marked increase in the incidence of necrotizing lung infections with a very high mortality associated with these strains. This report describes a fatal case of hospital-acquired necrotizing pneumonia caused by PVL-positive methicillin-susceptible S. aureus in a patient with a brain tumor.
Fulltext The Trend of Direct Medical Cost of Meliodiosis Patients in Kedah: A Retrospective Study from 2014 to 2017

Mardhiah K, Wan-Arfah N, Naing NN, Abu Hassan MR, Chan HK, Hasan H

Clinicoecon Outcomes Res, 2021;13:155-162.
PMID: 33732004 DOI: 10.2147/CEOR.S286283

Purpose: This study was conducted to determine the direct medical cost of treating melioidosis patients. The calculation was made according to the variables extracted from medical records.
Materials and Methods: Data collection was performed retrospectively on a total of 293 cases from Hospital Sultanah Bahiyah, Kedah, Malaysia. The data consisted of personal information, treatment history, and investigation findings, including blood results, USG abdomen results, and CT scan results. The site of culture and sensitivity were also obtained. The total direct medical cost was based on the antibiotics/treatments received by the patients, diagnostic test and investigations performed. The trend analysis used to see the pattern of costs from 2014 to 2017. All the costs were compared based on patients' status and duration of stay at the hospital using the independent t-test.
Results: The overall mean of direct medical cost for melioidosis amounted to US $233.61 (RM931.33). Overall, the finding confirms a huge reduction (44.7%) of direct medical cost from 2014 to 2017 (P = 0.001). From 2015 to 2016, there was a 19.1% reduction of direct medical cost (P>0.95), followed by a 38.8% reduction in costs from 2016 to 2017 (P = 0.019). In the case of the duration of stay, the mean of total direct medical cost among patients with ≥14 duration of stay was higher compared to those with <14 duration of stay (p < 0.001). There was no significant mean difference of direct medical cost between patients who were cured and died.
Conclusion: Despite the higher mortality of melioidosis cases compared to other infectious diseases, there is a limitation in the amount of published data on the management cost of melioidosis. The importance of cost in managing this disease should be underlined to perform a fully prepared management toward the disease.
Improved lovastatin production by inhibiting (+)-geodin biosynthesis in Aspergillus terreus

Hasan H, Abd Rahim MH, Campbell L, Carter D, Abbas A, Montoya A

N Biotechnol, 2019 Sep 25;52:19-24.
PMID: 30995533 DOI: 10.1016/j.nbt.2019.04.003

Lovastatin is widely prescribed to reduce elevated levels of cholesterol and prevent heart-related diseases. Cultivation of Aspergillus terreus (ATCC 20542) with carbohydrates or low-value feedstocks such as glycerol produces lovastatin as a secondary metabolite and (+)-geodin as a by-product. An A. terreus mutant strain was developed (gedCΔ) with a disrupted (+)-geodin biosynthesis pathway. The gedCΔ mutant was created by inserting the antibiotic marker hygromycin B (hyg) within the gedC gene that encodes emodin anthrone polyketide synthase (PKS), a primary gene responsible for initiating (+)-geodin biosynthesis. The effects of emodin anthrone PKS gene disruption on (+)-geodin and lovastatin biosynthesis and the production of the precursors acetyl-CoA and malonyl-CoA were investigated with cultures based on glycerol alone and in combination with lactose. The gedCΔ strain showed improved lovastatin production, particularly when cultivated on the glycerol-lactose mixture, increasing lovastatin production by 80% (113 mg/L) while simultaneously inhibiting (+)-geodin biosynthesis compared to the wild-type strain. This study thus shows that suppression of the (+)-geodin pathway increases lovastatin yield and demonstrates a practical approach of manipulating carbon flux by modulating enzyme activity.
Fulltext Thermostable Heptaplex PCR Assay for the Detection of Six Respiratory Bacterial Pathogens

Nik Zuraina NMN, Goni MD, Amalina KN, Hasan H, Mohamad S, Suraiya S

Diagnostics (Basel), 2021 Apr 22;11(5).
PMID: 33922299 DOI: 10.3390/diagnostics11050753

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.
Fulltext The association between pre-morbid conditions and respiratory tract manifestations amongst Malaysian Hajj pilgrims

Deris ZZ, Hasan H, Ab Wahab MS, Sulaiman SA, Naing NN, Othman NH

Trop Biomed, 2010 Aug;27(2):294-300.
PMID: 20962728 MyJurnal

In a very closed and overcrowding environment, influenza transmission during Hajj season is almost inevitable. The aim of this study was to determine the association between pre-morbid conditions and influenza-like illness (ILI) amongst Hajj pilgrims. A cross-sectional study was conducted amongst Malaysian Hajj pilgrims in year 2007. Survey forms were distributed at Madinatul-Hujjaj, Jeddah and Tabung Haji Clinic, Medina, Saudi Arabia where pilgrims stay on transit before returning to Malaysia. Allergic rhinitis was significantly associated with sore throat (p=0.047), longer duration of cough (p=0.017) and runny nose (p=0.016). Pilgrims who suffered from chronic obstructive pulmonary diseases (COPD) had significant association with longer duration of cough (p=0.041) and those with diabetes mellitus had significant association with longer duration of sore throat (p=0.048). Underlying asthma was significantly associated with severe influenza like illness requiring admission to hospital for further treatment of respiratory symptoms (p=0.016). Based on these findings, we suggest those with underlying asthma should be discouraged from participating in the hajj and they should seek early treatment if they develop respiratory symptoms.
Fulltext Evaluation of Total Female and Male Aedes aegypti Proteomes Reveals Significant Predictive Protein-Protein Interactions, Functional Ontologies, and Differentially Abundant Proteins

Shettima A, Joseph S, Ishak IH, Abdul Raiz SH, Abu Hasan H, Othman N

Insects, 2021 Aug 20;12(8).
PMID: 34442320 DOI: 10.3390/insects12080752

Aedes aegypti is a significant vector for many tropical and subtropical flavivirus diseases. Only the female mosquito transmits pathogens, while the male plays a vital role in mating and species continuity. This study explored the total proteomes of females and males based on the physiological and genetic differences of female and male mosquitoes. Protein extracts from mosquitoes were analysed using LC-ESI-MS/MS for protein identification, protein interaction network analysis, functional ontology enrichment, and differential protein abundance analyses. Protein identification revealed 422 and 682 proteins exclusive to males and females, respectively, with 608 common proteins found in both sexes. The most significant PPIs (<1.0 × 10-16) were for common proteins, followed by proteins exclusive to females (<1.0 × 10-16) and males (1.58 × 10-12). Significant functional enrichments were observed in the biological process, molecular function, and cellular component for the male and female proteins. The abundance of the proteins differed, with one protein showing an increase (elongation factor 1 α, EF1α) and two showing reductions (actin family) in females versus males. Overall, the study verified the total proteomes differences between male and female Ae. aegypti based on protein identification and interactions, functional ontologies, and differentially abundant proteins. Some of the identified proteins merit further investigation to elucidate their roles in blocking viral transmission.
Fulltext Combination of Molecular Networking and LC-MS/MS Profiling in Investigating the Interrelationships between the Antioxidant and Antimicrobial Properties of Curculigo latifolia

Mad Nasir N, Ezam Shah NS, Zainal NZ, Kassim NK, Faudzi SMM, Hasan H

Plants (Basel), 2021 Jul 21;10(8).
PMID: 34451533 DOI: 10.3390/plants10081488

Curculigo is a potent plant with a variety of traditional uses, such as anti-oxidant, anti-diabetic, anti-tumor, anti-bacterial, anti-cancer, anti-osteoporosis, and wound-healing. The comprehensive profiling of the Curculigolatifolia metabolome was carried out by generating a molecular network (MN) from Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) data to profile the methanol extract and correlating them with their antioxidant (2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), total phenolic contents (TPC), and β-carotene) and antimicrobial (disk-diffusion agar method, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC)) properties. The antioxidant capacity was observed to be significantly higher in the rhizome crude extract, with 18.10 ± 0.91 µg/mL DPPH activity, and a β-carotene bleaching result of 35.20%. For the antimicrobial activity, the leaf crude extract exhibited a strong Staphylococcus aureus and Salmonella choleraesuis (8-15 ± 3.0 mm) inhibition in the disk-diffusion agar. The leaf extract also exhibited maximum antibacterial activity against S. aureus (MIC = ±0.25 mg/mL, MBC = ±0.25 mg/mL) and S. choleraesuis (MIC = ±0.25 mg/mL, MBC = ±0.25 mg/mL). LC-MS/MS analysis and MN revealed norlignans and phenolic glycosides as major metabolites in the rhizome and leaf extracts of the negative mode (M - H)-. Fourteen known compounds were identified, and three unknown compounds were putatively identified in the rhizome extract, while ten known compounds and six unknown compounds were putatively identified in the leaf extract.
Screening of nematophagous-fungi from fresh faeces of grazing animals and soils

Farah Haziqah MT, Nur Hikmah AM, Mat Hasan H, Hamdan A, Nik Him NAII

Trop Biomed, 2019 Sep 01;36(3):687-693.
PMID: 33597490

An investigation was undertaken for screening and isolating nematophagous-fungi from the faecal samples of various grazing animals and soils in Malaysia. Total of 111 faeces and 50 soil samples were collected and the samples were cultured on 2% water agar plates. The growth of nematophagous-fungi was stimulated by sprinkling-baiting technique. The conidia of suspected nematophagous-fungi were inoculated on 2% water agar plates. All isolated were maintained on 2% cornmeal agar plates. Verticillium spp., Fusarium spp. and Arthrobotrys spp. were identified from the faecal and soil samples. 62.5% of the faecal samples and 100% of the soil samples were shown to be positive with nematophagous-fungi. This study highlights the present of nematophagous-fungi population in faecal and soil samples. Much study remains to be done to better understanding some fungi especially their mode of action and their predatory behaviour against parasitic nematodes.
Fulltext A Thermostabilized, One-Step PCR Assay for Simultaneous Detection of Klebsiella pneumoniae and Haemophilus influenzae

Khazani NA, Noor NZ, Yean Yean C, Hasan H, Suraiya S, Mohamad S

J Trop Med, 2017;2017:7210849.
PMID: 28386286 DOI: 10.1155/2017/7210849

Klebsiella pneumoniae and Haemophilus influenzae are 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 of K. pneumoniae and H. influenzae. The multiplex PCR assay was designed to amplify the php gene of K. pneumoniae (202 bp) and p6 gene of H. influenzae (582 bp). In addition, the specific primer to amplify glm gene of Helicobacter 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 the Q(10) method. 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 the Q10 accelerated stability was found to be 3.02 months. A cold-chain-free, PCR assay for easy, rapid, and simultaneous detection of K. pneumoniae and H. influenzae was successfully developed in this study.
Overexpression of acetyl-CoA carboxylase in Aspergillus terreus to increase lovastatin production

Hasan H, Abd Rahim MH, Campbell L, Carter D, Abbas A, Montoya A

N Biotechnol, 2018 Sep 25;44:64-71.
PMID: 29727712 DOI: 10.1016/j.nbt.2018.04.008

The present work describes the application of homologous recombination techniques in a wild-type Aspergillus terreus (ATCC 20542) strain to increase the flow of precursors towards the lovastatin biosynthesis pathway. A new strain was generated to overexpress acetyl-CoA carboxylase (ACCase) by replacing the native ACCase promoter with a strong constitutive PadhA promoter from Aspergillus nidulans. Glycerol and a mixture of lactose and glycerol were used independently as the carbon feedstock to determine the degree of response by the A. terreus strains towards the production of acetyl-CoA, and malonyl-CoA. The new strain increased the levels of malonyl-CoA and acetyl-CoA by 240% and 14%, respectively, compared to the wild-type strain. As a result, lovastatin production was increased by 40% and (+)-geodin was decreased by 31% using the new strain. This study shows for the first time that the metabolism of Aspergillus terreus can be manipulated to attain higher levels of precursors and valuable secondary metabolites.
Fulltext High frequency of Haemophilus influenzae associated with respiratory tract infections among Malaysian Hajj pilgrims

Nik Zuraina NMN, Sarimah A, Suharni M, Hasan H, Suraiya S

J Infect Public Health, 2018 08 07;11(6):878-883.
PMID: 30097415 DOI: 10.1016/j.jiph.2018.07.010

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.

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