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Fulltext Assessing product adulteration of Eurycoma longifolia (Tongkat Ali) herbal medicinal product using DNA barcoding and HPLC analysis

Abubakar BM, Salleh FM, Shamsir Omar MS, Wagiran A

Pharm Biol, 2018 Dec;56(1):368-377.
PMID: 30058427 DOI: 10.1080/13880209.2018.1479869

CONTEXT: Eurycoma longifolia Jack (Simaroubaceae) commonly known as Tongkat Ali is one of the most important plants in Malaysia. The plant extracts (particularly roots) are widely used for the treatment of cough and fever besides having antimalarial, antidiabetic, anticancer and aphrodisiac activities.
OBJECTIVES: This study assesses the extent of adulteration of E. longifolia herbal medicinal products (HMPs) using DNA barcoding validated by HPLC analysis.
MATERIALS AND METHODS: Chloroplastic rbcL and nuclear ITS2 barcode regions were used in the present study. The sequences generated from E. longifolia HMPs were compared to sequences in the GenBank using MEGABLAST to verify their taxonomic identity. These results were verified by neighbor-joining tree analysis in which branches of unknown specimen are compared to the reference sequences established from this study and other retrieved from the GenBank. The HMPs were also analysed using HPLC analysis for the presence of eurycomanone bioactive marker.
RESULTS: Identification using DNA barcoding revealed that 37% of the tested HMPs were authentic while 27% were adulterated with the ITS2 barcode region proven to be the ideal marker. The validation of the authenticity using HPLC analysis showed a situation in which a species which was identified as authentic was found not to contain the expected chemical compound.
DISCUSSION AND CONCLUSIONS: DNA barcoding should be used as the first screening step for testing of HMPs raw materials. However, integration of DNA barcoding with HPLC analysis will help to provide detailed knowledge about the safety and efficacy of the HMPs.
Fulltext Soil-transmitted helminth infections and associated risk factors in three Orang Asli tribes in Peninsular Malaysia

Anuar TS, Salleh FM, Moktar N

Sci Rep, 2014;4:4101.
PMID: 24525479 DOI: 10.1038/srep04101

Currently, information on prevalence of soil-transmitted helminth (STH) infections among different tribes of Orang Asli (aboriginal) is scarce in Malaysia. The present study is a cross-sectional study aimed at determining the factors associated with the prevalence of STH infections among the Proto-Malay, Negrito and Senoi tribes. Faecal samples were collected from 500 participants and socioeconomic data was collected via pre-tested questionnaire. All samples were processed using formalin-ether sedimentation and Wheatley's trichrome staining. Trichuris trichiura (57%) was the most common STH seen among the participants, followed by Ascaris lumbricoides (23.8%) and hookworm (7.4%). Trichuriasis and ascariasis showed an age-dependency relationship; significantly higher rates were observed among Senois who aged <15 years. Likewise, Negritos also showed an age-dependency association with ascariasis affecting mainly the under 15 years old individuals. Multivariate logistic regression model indicated the following predictors of trichuriasis among these communities; being aged <15 years, consuming raw vegetables, belonging to a large household members (≥8) and earning low household income (
Fulltext Different clinical outcomes of Entamoeba histolytica in Malaysia: does genetic diversity exist?

Anuar TS, Al-Mekhlafi HM, Abdul Ghani MK, Azreen SN, Salleh FM, Ghazali N, et al.

Korean J Parasitol, 2013 Apr;51(2):231-6.
PMID: 23710093 DOI: 10.3347/kjp.2013.51.2.231

The present study was conducted to investigate the clinical outcomes of Entamoeba histolytica infection in symptomatic and asymptomatic Orang Asli (aborigine) communities in Malaysia. Examination was performed on 500 stool samples obtained from Orang Asli communities in 3 different states using formalin-ether concentration, trichrome staining, and single-round PCR techniques. Out of 500 stool samples, single infection of E. histolytica, Entamoeba dispar, and Entamoeba moshkovskii was identified in 3.2%, 13.4%, and 1%, respectively. In addition, 10 samples had mixed infections with E. histolytica and E. dispar. Six samples containing E. dispar were also positive for E. moshkovskii, and only 2 samples had E. histolytica in association with E. dispar and E. moshkovskii. Seventeen E. histolytica-positive samples were from symptomatic subjects, whereas the remaining 11 samples came from asymptomatic subjects. These findings suggest a predominant distribution of pathogenic potential of E. histolytica strains in this community. Therefore, further studies on genotyping of E. histolytica is required, to find out association between E. histolytica genotype and the outcome of the infection.
First molecular identification of Entamoeba moshkovskii in Malaysia

Anuar TS, Al-Mekhlafi HM, Ghani MK, Azreen SN, Salleh FM, Ghazali N, et al.

Parasitology, 2012 Oct;139(12):1521-5.
PMID: 22939193 DOI: 10.1017/S0031182012001485

Entamoeba moshkovskii and Entamoeba dispar are microscopically indistinguishable from the pathogenic species Entamoeba histolytica. Although sporadic cases of human infection with E. moshkovskii have been reported, the amoeba is still considered primarily as a free-living amoeba. A cross-sectional study was carried out among Orang Asli communities in 3 different states of Peninsular Malaysia. Fecal samples were examined by formalin-ether sedimentation and trichrome staining techniques and then single-round PCR assay was used to detect E. moshkovskii. Out of 500 fecal samples examined microscopically, 93 (18·6%) samples were positive for E. histolytica/E. dispar/E. moshkovskii complex cysts and/or trophozoites. PCR products were detected in 106 fecal samples. E. moshkovskii isolates were detected in 13 (12·3%) fecal samples. Of the 13 E. moshkovskii-positive samples, 5 were of single isolation of E. moshkovskii, 6 were also positive for E. dispar, and only 2 samples were positive for E. dispar and E. histolytica. Moreover, 3 E. moshkovskii-positive samples were collected from symptomatic individuals while the remaining 10 samples were from asymptomatic subjects. This is the first report on the identification of E. moshkovskii in Malaysia. Further studies are needed to confirm the pathogenicity of E. moshkovskii infection and determine the epidemiology among Orang Asli communities in Malaysia.
Fulltext Blastocystis infection in Malaysia: evidence of waterborne and human-to-human transmissions among the Proto-Malay, Negrito and Senoi tribes of Orang Asli

Anuar TS, Ghani MK, Azreen SN, Salleh FM, Moktar N

Parasit Vectors, 2013;6:40.
PMID: 23433099 DOI: 10.1186/1756-3305-6-40

Blastocystis has been described as the most common intestinal parasite in humans and has an increased impact on public health. However, the transmission of this parasite has not been conclusively determined.
Evaluation of formalin-ether sedimentation and trichrome staining techniques: its effectiveness in detecting Entamoeba histolytica/dispar/moshkovskii in stool samples

Anuar TS, Al-Mekhlafi HM, Abdul Ghani MK, Abu Bakar E, Azreen SN, Salleh FM, et al.

J Microbiol Methods, 2013 Mar;92(3):344-8.
PMID: 23361047 DOI: 10.1016/j.mimet.2013.01.010

This study was conducted to evaluate two routinely microscopic diagnostic methods in comparison with single-round PCR assay as the reference technique to detect Entamoeba histolytica/dispar/moshkovskii. Examination was performed on 500 stool samples obtained from Orang Asli communities in different states of Malaysia using formalin-ether sedimentation, trichrome staining and single-round PCR techniques. Ninety-three stool samples were detected E. histolytica/dispar/moshkovskii positive by routine microscopy, while single-round PCR detected 106 positive samples. Additional positives detected by PCR assay were eventually confirmed to be negative by both microscopic techniques. Detection rate of E. histolytica/dispar/moshkovskii was highest in combination techniques (18.6%), followed by trichrome staining (13.4%) and formalin-ether sedimentation (11.2%) techniques. Single-round PCR detected 21.2% of the stool samples. The sensitivity and specificity of formalin-ether sedimentation and trichrome staining techniques compared to the reference technique were 31.1% (95% CI: 29.0-36.0) and 94.2% (95% CI: 89.8-98.9), and 53.8% (95% CI: 46.0-76.2) and 97.5% (95% CI: 92.8-99.1), respectively. However, the sensitivity [59.4% (95% CI: 48.9-78.5)] of the method increased when both techniques were performed together, but the specificity decreased to 92.4% (95% CI: 81.0-98.0). The agreement between the reference technique, trichrome staining and combination techniques were statistically significant by Kappa statistics (trichrome staining: K = 0.592, p < 0.05; combination techniques: K = 0.543, p < 0.05). Hence, the combination technique is recommended to be used as a screening method in the diagnosis of E. histolytica/dispar/moshkovskii infections either for clinical or epidemiological study.
Fulltext Human giardiasis in Malaysia: correlation between the presence of clinical manifestation and Giardia intestinalis assemblage

Anuar TS, Moktar N, Salleh FM, Al-Mekhlafi HM

Southeast Asian J. Trop. Med. Public Health, 2015 Sep;46(5):835-43.
PMID: 26863854

Clinical manifestations of giardiasis vary from asymptomatic infection to chronic diarrhea. A total of 611 stool samples from Aboriginal participants residing in Jelebu, Gerik and Temerloh States, Malaysia, ages 2 to 74 years were screened for Giardia intestinalis using microscopic examination and sequence analysis of a fragment of nested-PCR amplified triosephosphate isomerase (tpi) gene. Demographic data was collected through a structured questionnaire. tpi was successfully amplified from 98/110 samples microscopically positive for G. intestinalis, with 62 and 36 belonging to assemblage A and B, respectively. There is a significant correlation between assemblage A and symptomatic infection only in participants of < 15 years of age. In the other age group, host factors may have more effects on the presence of clinical signs and symptoms than G. intestinalis assemblage types.
Fulltext Prevalence of Dientamoeba fragilis among an Orang Asli population in rural Malaysia

Anuar TS, Hasim L, Moktar N, Salleh FM, Al-Mekhlafi HM

Southeast Asian J. Trop. Med. Public Health, 2015 Sep;46(5):844-9.
PMID: 26863855

Dientamoeba fragilis is a trichomonad parasite that can infect the gastrointestinal tract of humans causing gastrointestinal disease. Little is known about its epidemiology. We evaluated the prevalence of D. fragilis by conducting a cross sectional study of an Orang Asli population in rural Malaysia. We examined stool samples from 150 participants for D. fragilis using Wheatley's trichrome stain and collected demographic data from each participant using a structured questionnaire. Five participants (3.3%) had D. fragilis in their stool; four of these were aged < 15 years; 3 were male and 2 were female. All participants with positive stool sample for D. fragilis were symptomatic; 3 had diarrhea and 2 had other gastrointestinal symptoms. D. fragilis is present in the study population. Further studies are needed to determine the virulence, pathogenicity and mode of transmission of D. fragilis in the study population.
Molecular epidemiology of amoebiasis in Malaysia: highlighting the different risk factors of Entamoeba histolytica and Entamoeba dispar infections among Orang Asli communities

Anuar TS, Al-Mekhlafi HM, Abdul Ghani MK, Abu Bakar E, Azreen SN, Salleh FM, et al.

Int J Parasitol, 2012 Dec;42(13-14):1165-75.
PMID: 23123168 DOI: 10.1016/j.ijpara.2012.10.003

Currently, species-specific information on Entamoeba infections is unavailable in Malaysia and is restricted worldwide due to the re-description of pathogenic Entamoeba histolytica and non-pathogenic Entamoeba dispar and Entamoeba moshkovskii. Therefore, this cross-sectional study was conducted to provide the first known documented data on the true prevalence of these three species in western Malaysia using a molecular method. Another aim of this study was to determine the association of potential risk factors associated with each Entamoeba sp. A total of 500 stool samples from three Orang Asli tribes were randomly collected. The overall prevalence of E. histolytica, E. dispar and E. moshkovskii determined by microscopy was 18.6% (93/500). Molecular analysis revealed that while most Entamoeba-positive individuals were infected with E. dispar (13.4%), followed by E. histolytica (3.2%) and E. moshkovskii (1.0%), the present findings show low prevalence rates of mixed infections with E. histolytica and E. dispar (2%), E. dispar and E. moshkovskii (1.2%) and association infections of E. histolytica, E. dispar and E. moshkovskii (0.4%). Logistical regression analysis indicates that the dynamics of the transmission of the three Entamoeba spp. was different. Of six statistically significant variables observed in the univariate analysis, three were retained as significant risk factors for E. histolytica infection in the logistical regression model. These factors were (i) not washing hands after playing with soil or gardening (Odds ratio (OR)=4.7; 95% confidence level (CI)=1.38, 16.14; P=0.013), (ii) indiscriminate defecation in the river or bush (OR=5.7; 95% CI=1.46, 21.95; P=0.012) and (iii) close contact with domestic animals (OR=5.4; 95% CI=1.36, 2.51; P=0.017). However, subjects with family members who were infected with E. histolytica/E. dispar/E. moshkovskii (OR=3.8; 95 CI=2.11, 6.86; P<0.001) and those who consumed raw vegetables (OR=1.8; 95% CI=1.01, 3.23; P=0.047) were more likely to be infected with E. dispar. On the other hand, no associated factor was identified with E. moshkovskii infection. Nevertheless, diarrhoea (P=0.002) and other gastroenteritis symptoms (P<0.001) were only associated with E. histolytica infection. The present study provides new insight into the distribution and risk factors of E. histolytica, E. dispar and E. moshkovskii infections among Orang Asli communities in Malaysia. Identifying the different risk factors of E. histolytica and E. dispar infections will help in the planning specific strategies in the control and prevention of each infection in the communities. Moreover, it emphasises the need for molecular methods to determine the species-specific prevalence of Entamoeba spp.
Giardiasis among different tribes of Orang Asli in Malaysia: highlighting the presence of other family members infected with Giardia intestinalis as a main risk factor

Anuar TS, Al-Mekhlafi HM, Ghani MK, Osman E, Yasin AM, Nordin A, et al.

Int J Parasitol, 2012 Aug;42(9):871-80.
PMID: 22846786 DOI: 10.1016/j.ijpara.2012.07.003

The flagellate protozoan parasite, Giardia intestinalis, is widely distributed throughout the world with a high prevalence in developing countries in the tropics and subtropics, including Malaysia. Approximately 200 million people are infected with the parasite globally, with 500,000 new cases reported annually. This cross-sectional study was conducted among three tribes of Orang Asli communities in Selangor, Perak and Pahang states of Malaysia. The main objective was to determine the prevalence of and risk factors for giardiasis. Stool samples were collected from 500 individuals aged between 2 and 74 years (males=219, females=281). The samples were examined with formalin-ether sedimentation and trichrome staining techniques. Socioeconomic data were collected through a pre-tested questionnaire. The overall prevalence of giardiasis was 20.0% with the highest prevalence in the Proto-Malays (33.3%) followed by Negritos (20.1%) and Senois (10.4%). The positive cases showed a decrease with increasing age and most of the positive cases were observed in individuals less than 24 years old. Males had significantly higher prevalence than females (χ(2)=5.283, P=0.022). Logistic regression analysis of the overall population studied and the Senoi tribe confirmed that being a child aged less than 15 years, being male, the consumption of raw vegetables and the presence of other family members infected with G. intestinalis were the main risk factors for giardiasis. The presence of other family members infected with G. intestinalis was the only risk factor highlighted in the Proto-Malay and Negrito tribes. Diarrhoea was significantly associated with giardiasis. However, the cause and effect relationship has yet to be determined. Thus, screening family members and treating the infected individuals are the main strategies that should be adopted by the public health authority in combating this infection in Orang Asli communities as well as health education regarding good personal and food hygiene practises.
Fulltext Molecular epidemiology of giardiasis among Orang Asli in Malaysia: application of the triosephosphate isomerase gene

Anuar TS, Azreen SN, Salleh FM, Moktar N

BMC Infect Dis, 2014 Feb 12;14:78.
PMID: 24520940 DOI: 10.1186/1471-2334-14-78

BACKGROUND: Giardia duodenalis is a flagellate parasite which has been considered the most common protozoa infecting human worldwide. Molecular characterization of G. duodenalis isolates have revealed the existence of eight groups (Assemblage A to H) which differ in their host distribution. Assemblages A and B are found in humans and in many other mammals.
METHODS: This cross-sectional study was conducted to identify assemblage's related risk factors of G. duodenalis among Orang Asli in Malaysia. Stool samples were collected from 611 individuals aged between 2 and 74 years old of whom 266 were males and 345 were females. Socioeconomic data were collected through a pre-tested questionnaire. All stool samples were processed with formalin-ether sedimentation and Wheatley's trichrome staining techniques for the primary identification of G. duodenalis. Molecular identification was carried out by the amplification of a triosephosphate isomerase gene using nested-PCR assay.
RESULTS: Sixty-two samples (10.2%) were identified as assemblage A and 36 (5.9%) were assemblage B. Risk analysis based on the detected assemblages using univariate and logistic regression analyses identified subjects who have close contact with household pets i.e. dogs and cats (OR = 2.60; 95% CI = 1.42, 4.78; P = 0.002) was found to be significant predictor for assemblage A. On the other hand, there were three significant risk factors caused by assemblage B: (i) children ≤15 years old (OR = 2.33; 95% CI = 1.11, 4.87; P = 0.025), (ii) consuming raw vegetables (OR = 2.82; 95% CI = 1.27, 6.26; P = 0.011) and (iii) the presence of other family members infected with giardiasis (OR = 6.31; 95% CI = 2.99, 13.31; P
Fulltext An international, stepped wedge, cluster-randomized trial investigating the 0/1-h algorithm in suspected acute coronary syndrome in Asia: the rational of the DROP-Asian ACS study

Inoue K, Chieh JTW, Yeh LC, Chiang SJ, Phrommintikul A, Suwanasom P, et al.

Trials, 2022 Dec 07;23(1):986.
PMID: 36476401 DOI: 10.1186/s13063-022-06907-4

BACKGROUND: More than half of the world's population lives in Asia. With current life expectancies in Asian countries, the burden of cardiovascular disease is increasing exponentially. Overcrowding in the emergency departments (ED) has become a public health problem. Since 2015, the European Society of Cardiology recommends the use of a 0/1-h algorithm based on high-sensitivity cardiac troponin (hs-cTn) for rapid triage of patients with suspected non-ST elevation acute coronary syndrome (NSTE-ACS). However, these algorithms are currently not recommended by Asian guidelines due to the lack of suitable data.
METHODS: The DROP-Asian ACS is a prospective, stepped wedge, cluster-randomized trial enrolling 4260 participants presenting with chest pain to the ED of 12 acute care hospitals in five Asian countries (UMIN; 000042461). Consecutive patients presenting with suspected acute coronary syndrome between July 2022 and Apr 2024 were included. Initially, all clusters will apply "usual care" according to local standard operating procedures including hs-cTnT but not the 0/1-h algorithm. The primary outcome is the incidence of major adverse cardiac events (MACE), the composite of all-cause death, myocardial infarction, unstable angina, or unplanned revascularization within 30 days. The difference in MACE (with one-sided 95% CI) was estimated to evaluate non-inferiority. The non-inferiority margin was prespecified at 1.5%. Secondary efficacy outcomes include costs for healthcare resources and duration of stay in ED.
CONCLUSIONS: This study provides important evidence concerning the safety and efficacy of the 0/1-h algorithm in Asian countries and may help to reduce congestion of the ED as well as medical costs.
Fulltext Correction: An international, stepped wedge, cluster-randomized trial investigating the 0/1-h algorithm in suspected acute coronary syndrome in Asia: the rational of the DROP-Asian ACS study

Inoue K, Chieh JTW, Yeh LC, Chiang SJ, Phrommintikul A, Suwanasom P, et al.

Trials, 2023 May 17;24(1):335.
PMID: 37198646 DOI: 10.1186/s13063-023-07342-9
Using wintergreen oil for mounting mosquito larvae: a safer alternative to xylene

Koay JB, Natasya NN, Nashithatul M, Ihsanuddin R, Salleh FM, Azil AH

Biotech Histochem, 2016;91(1):63-70.
PMID: 26528914 DOI: 10.3109/10520295.2015.1074286

Permanent mounting of fourth instar mosquito larvae is essential for identifying Aedes spp. This procedure requires extensive exposure to xylene, a clearing agent in the mounting process. We investigated wintergreen oil as a substitute for xylene. Five hundred larvae were mounted on slides to evaluate shrinkage or expansion of specimens after clearing using xylene or wintergreen oil. We examined the ventral brush and siphonal hair tufts for species identification and for preservation of morphological characteristics after clearing specimens in xylene or wintergreen oil. Shrinkage of the length of whole larvae and width of the head, thorax and abdomen after mounting was significantly greater after clearing with xylene than with wintergreen oil. The length of the comb scale nearest the ventral brush was similar for both clearing agents. The clarity of the specimens after mounting was improved by clearing with wintergreen oil, but the integrity of the ventral brush and siphonal hair tufts were similar for both clearing agents.
The complete mitochondrial genome data of the Common Rose butterfly, Pachliopta aristolochiae (Lepidoptera, Papilionoidea, Papilionidae) from Malaysia

Miga M, Jahari PNS, Vei Siang C, Kamarudin KR, Shamsir MS, Tokiman L, et al.

Data Brief, 2022 Feb;40:107740.
PMID: 35141362 DOI: 10.1016/j.dib.2021.107740

Here, we present the complete mitochondrial genome of Pachliopta aristolochiae, a Common Rose butterfly from Malaysia. The sequence was generated using Illumina NovaSeq 6000 sequencing platform. The mitogenome is 15,235bp long, consisting of 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and two D-loop regions. The total base composition was (81.6%), with A (39.3%), T (42.3%), C (11.0%) and G (7.3%). The gene order of the three tRNAs was trnM-trnI-trnQ, which differs from the ancestral insect gene order trnI-trnQ-trnM. Phylogenetic tree analysis revealed that the sequenced Pachliopta aristolochiae in this data is closely related to Losaria neptunus (NC 037868), with highly supported ML and BI analysis. The data presented in this work can provide useful resources for other researchers to study deeper into the phylogenetic relationships of Lepidoptera and the diversification of the Pachliopta species. Also, as one of the bioindicator species, this data can be used to assess environmental changes in the terrestrial and aquatic ecosystem via enviromental DNA approahes. The mitogenome of Pachliopta aristolochiae is available in GenBank under the accession number MZ781228.
Fulltext Characterization of the complete mitogenome data of Ischyja marapok (Lepidoptera: Noctuoidea: Erebidae) from Malaysia

Miga M, Jahari PNS, Parimannan S, Rajandas H, Abdul-Latiff MAB, Wei YJ, et al.

Data Brief, 2023 Jun;48:109253.
PMID: 37383759 DOI: 10.1016/j.dib.2023.109253

Ischyja marapok is a moth species from the genus Ischyja, a member of the Lepidoptera family, Erebidae. Due to their wide variation, this family constitutes the largest described species, however, the mitogenome dataset on the genus Ischyja is scarce. Hence, the mitochondrial genome dataset of Ischyja marapok from Malaysia was completely sequenced using the next-generation sequencing technology, Illumina NovaSeq 6000 and analyzed. The mitogenome has a sequence length of 15,421 bp, consisting of 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (rRNAs) and a control region. The mitogenome is A + T biased (80.6%), with the base composition of A (39.2%), T (41.4%), C (11.9%) and G (7.5%). Among the 13 PCGs, 12 were initiated by the standard ATN codon, except for COX1 which utilizes the CGA start codon. Two PCGs were terminated with an incomplete stop codon T, while others ended with a TAA codon. Phylogenetic tree analyses showed that the sequenced I. marapok resides within the Erebinae subfamily and is closely related to Ischyja manlia (MW664367) with high bootstrap support and posterior probabilities. This dataset presented the mitogenome data of I. marapok from Malaysia, which is valuable for further research of their phylogeny and the diversification of the Ischyja genus. Also, this dataset can be implemented and used as references to assess environmental changes in the terrestrial ecosystem via environmental DNA approaches. The mitogenome of I. marapok is available in GenBank under the accession number ON165249.
Severity of Malaria cases reported in urban and rural hospitals in Malaysia

Nimir AR, Isa NH, Eugene CB, Ghauth IM, Salleh FM, Rahman RA

Southeast Asian J. Trop. Med. Public Health, 2006 Sep;37(5):831-7.
PMID: 17333723

The age distribution, types of infection and clinical patterns of malaria were compared in patients admitted to an urban and a rural hospital. Analysis of the cases seen in urban setting characteristically indicated a relatively low transmission rate of the disease, whereas the mean inoculation rate in patients from the rural hospital was found to be at least twenty folds higher. Plasmodium vivax was the predominant causative species in the urban hospital (p = 0.01), infecting mostly adult (p = 0.001) males (p = 0.01). The geometric mean parasite count at 3432/microl among the urban patients was significantly higher than that in the rural patients at 1422/microl (p = 0.04). Coma and death were more common among the cases seen in the urban hospital (p = 0.003), while severe anemia was the significant complication in the rural setting. Overall, the provisional diagnosis of malaria was relatively low in the urban hospital (p = 0.02). The results from this study highlighted the need to define the extent of malaria in urban areas. This report attempts to identify the non-climatic determinants of the infection and, furthermore, to provide a more informed basis to describe the burden of the disease.
Fulltext Molecular epidemiology of blastocystosis in Malaysia: does seasonal variation play an important role in determining the distribution and risk factors of Blastocystis subtype infections in the Aboriginal community?

Noradilah SA, Moktar N, Anuar TS, Lee IL, Salleh FM, Manap SNAA, et al.

Parasit Vectors, 2017 Jul 31;10(1):360.
PMID: 28760145 DOI: 10.1186/s13071-017-2294-2

BACKGROUND: Alternating wet and dry seasons may play an important role in the acquisition and distribution of Blastocystis subtype infection in the tropics. This cross-sectional study was therefore conducted to provide the prevalence of Blastocystis and to determine the potential risk factors associated with each subtype during the wet and dry seasons in the Aboriginal community, Pahang, Malaysia.
METHODS: A total of 473 faecal samples were collected: 256 (54.1%) and 217 (45.9%) samples were obtained during the wet (October-November 2014) and the dry season (June 2015), respectively. All fresh faecal samples were subjected to molecular analysis for subtype and allele identification.
RESULTS: Of the 473 samples, 42.6% and 37.8% were positive for Blastocystis ST1, ST2, ST3 and ST4 during wet and dry seasons, respectively. Prevalence of Blastocystis ST1 was significantly higher during the wet season compared to the dry season (Z = 2.146, P
Occurrence of Blastocystis sp. in water catchments at Malay villages and Aboriginal settlement during wet and dry seasons in Peninsular Malaysia

Noradilah SA, Lee IL, Anuar TS, Salleh FM, Abdul Manap SN, Mohd Mohtar NS, et al.

PeerJ, 2016;4:e2541.
PMID: 27761331

In the tropics, there are too few studies on isolation of Blastocystis sp. subtypes from water sources; in addition, there is also an absence of reported studies on the occurrence of Blastocystis sp. subtypes in water during different seasons. Therefore, this study was aimed to determine the occurrence of Blastocystis sp. subtypes in river water and other water sources that drained aboriginal vicinity of highly endemic intestinal parasitic infections during wet and dry seasons. Water samples were collected from six sampling points of Sungai Krau (K1-K6) and a point at Sungai Lompat (K7) and other water sources around the aboriginal villages. The water samples were collected during both seasons, wet and dry seasons. Filtration of the water samples were carried out using a flatbed membrane filtration system. The extracted DNA from concentrated water sediment was subjected to single round polymerase chain reaction and positive PCR products were subjected to sequencing. All samples were also subjected to filtration and cultured on membrane lactose glucuronide agar for the detection of faecal coliforms. During wet season, Blastocystis sp. ST1, ST2 and ST3 were detected in river water samples. Blastocystis sp. ST3 occurrence was sustained in the river water samples during dry season. However Blastocystis sp. ST1 and ST2 were absent during dry season. Water samples collected from various water sources showed contaminations of Blastocystis sp. ST1, ST2, ST3 and ST4, during wet season and Blastocystis sp. ST1, ST3, ST8 and ST10 during dry season. Water collected from all river sampling points during both seasons showed growth of Escherichia coli and Enterobacter aerogenes, indicating faecal contamination. In this study, Blastocystis sp. ST3 is suggested as the most robust and resistant subtype able to survive in any adverse environmental condition. Restriction and control of human and animal faecal contaminations to the river and other water sources shall prevent the transmission of Blastocystis sp. to humans and animals in this aboriginal community.
Fulltext The prevalence of Enterobius vermicularis amongst primary school children in Pulau Ketam, Selangor--1988

Oothuman P, Rampal L, Omar B, Marwsi MA, Mohammod CG, Jeffery J, et al.

Med J Malaysia, 1989 Dec;44(4):312-6.
PMID: 2520040

A survey was conducted to investigate the prevalence of Enterobius vermicularis in 1352 Chinese children between the ages of 6-12 years from the three National Type Chinese Primary Schools in Pulau Ketam in 1988, using the scotch-tape technique on three successive days. The overall prevalence was high (56.88%). The prevalence in Sin Bin School was significantly lower (50.86%) compared to the other two schools (56.60% and 61.04%). Prevalence was significantly higher amongst the 6-7 years age group (61.61%-70.18%) compared to other age groups (47.9%-59.29%). There was no difference in the prevalence between the boys and girls being 56.15% and 57.55% respectively.

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