Displaying publications 1 - 20 of 29 in total

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  1. 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 (
  2. Shahrul Anuar T, M Al-Mekhlafi H, Md Salleh F, Moktar N
    PLoS One, 2013;8(8):e71870.
    PMID: 24014078 DOI: 10.1371/journal.pone.0071870
    Studies on microsporidial infection mostly focus on immunodeficiency or immunosuppressive individuals. Therefore, this cross-sectional study describes the prevalence and risk factors of microsporidiosis among asymptomatic individuals in Malaysia.
  3. Salleh FM, Anuar TS, Yasin AM, Moktar N
    J Microbiol Methods, 2012 Oct;91(1):174-8.
    PMID: 22986100 DOI: 10.1016/j.mimet.2012.08.004
    Permanent staining of faecal smears by Wheatley's trichrome technique has been used by many scientists for the detection of parasites in the past and it was found to be highly sensitive. This study was conducted to evaluate the use of Wintergreen oil in comparison with xylene in Wheatley's trichrome staining technique, as the reference technique. In a blind comparison study, 500 collected faecal samples from aboriginal communities were examined. Wintergreen oil was found to be more superior than xylene as a clearing agent in the Wheatley's trichrome staining of polyvinyl alcohol-fixed faecal smears for the identification of intestinal protozoa. Elimination of toxic, carcinogenic, and fire hazards makes Wintergreen oil the preferred choice in routine parasitology examinations.
  4. Moktar NM, Yusof HM, Yahaya NH, Muhamad R, Das S
    Clin Ter, 2010;161(1):25-8.
    PMID: 20393674
    AIMS: The mRNA level for interleukin-6 (IL-6) is an important marker of osteoarthritis (OA). The present study aimed to investigate the level of IL-6 mRNA in the cartilage of OA knee while comparing it to the normal cartilage obtained from the same patient.
    MATERIALS AND METHODS: A total of 21 patients who underwent total knee replacement were recruited for this study. Sectioning of the destructive cartilage was performed in the medial part of the proximal tibiofemoral cartilage. The unaffected lateral part of the knee in the same patient, served as a control. The mRNA level for IL-6 was assessed using LightCycler 2.0 quantitative real-time polymerase chain reaction (qRT-PCR). actin mRNA was used as an endogenous control.
    RESULTS: Twelve out of 21 patients (57.1%) exhibited up regulation of IL-6 mRNA in the OA cartilage as compared to the normal cartilage. The rest of the patients (42.9%) showed down regulation of IL-6 mRNA. The statistical analysis showed there was insignificant level of IL-6 mRNA in the OA (1.91 +/- 0.45) as compared to the normal cartilage (1.13 +/- 0.44) (p > 0.05). The inter-individual variation in the level of IL-6 mRNA in the cartilage of idiopathic knee was in accordance with previous findings.
    CONCLUSIONS: These observations suggest IL-6 could also act as a catabolic agent in some patients or its expression might be influenced by other cytokines.
    Study site: Pusat Perubatan Universiti Kebangsaan Malaysia (PPUKM), Kuala Lumpur, Malaysia
  5. Anuar TS, Moktar N, Salleh FM, Al-Mekhlafi HM
    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.
  6. Ashikin A, Al-Mekhlafi HM, Moktar N, Anuar TS
    Parasitol Int, 2017 Apr;66(2):163-165.
    PMID: 28115231 DOI: 10.1016/j.parint.2017.01.013
    Most studies of opportunistic infections focus on immunocompromised patients. However, there is a lack of information on microsporidiosis in healthy people (immunocompetent) worldwide. This study aimed to detect and identify microsporidia species in immunocompetent Orang Asli living in Pahang, Malaysia. Orang Asli is a collective term for a group of indigenous people that usually reside in the interior regions of Peninsular Malaysia. They comprise about 0.7% of the total population in Malaysia and 76% of them lived below the poverty line i.e., poor housing conditions with the lack of access to safe drinking water and adequate sanitation, contaminated environment, high illiteracy rate and unhygienic practices by these people. Stool samples were collected from 209 Orang Asli and analyzed for detecting the presence of Enterocytozoon bieneusi and Encephalitozoon intestinalis by polymerase chain reaction assay targeting small subunit ribosomal RNA gene. E. bieneusi was detected in 8 individuals (3.83%). This infection was commonly found in males than females (5.2% vs. 2.7%). All infected Orang Asli were adults, with a mean age of 44years. Diarrhea and other gastrointestinal symptoms were reported in one case (12.5%) among individuals infected with this species. These findings clearly show that exposure to E. bieneusi may actually be common than reported. The accurate detection and identification of microsporidian species by molecular technique will improve therapy, clinical manifestations and prognosis of this infection, as no antiparasitic therapy has been approved for E. bieneusi. It is hoped that these findings will allow the formulation of better health management and disease prevention advisories, and improvement in the standards of health in similar communities.
  7. Mohammad NA, Al-Mekhlafi HM, Moktar N, Anuar TS
    Asian Pac J Trop Med, 2017 May;10(5):491-497.
    PMID: 28647187 DOI: 10.1016/j.apjtm.2017.05.001
    OBJECTIVES: To determine the prevalence and risk factors of Blastocystis among underprivileged communities living in rural Malaysia.

    METHODS: This cross-sectional study was conducted among 253 participants aged between 1 and 85 years. Stool samples were examined using Wheatley's trichrome stain after in-vitro cultivation in Jones' medium to detect the presence of Blastocystis. Information pertaining to the demography, socioeconomic and environment were collected using pre-validated questionnaires.

    RESULTS: The total prevalence of Blastocystis infection was 40.7%. The multiple logistic regression analysis revealed that age ≥15 years (OR = 2.72; 95% CI = 1.47-5.04) and presence of infected family members (OR = 8.56; 95% CI = 4.47-16.38) were the significant risk factors associated with blastocystosis in these communities.

    CONCLUSIONS: Blastocystosis is revealed through this study to be still prevalent among Orang Asli communities in rural Malaysia. The two main approaches that should be implemented by the public health authority in battling this infection would be the screening of other family members and giving treatment to the infected individuals. Moreover, it is imperative for health education on good personal and food hygiene practices are provided in order to reduce the morbidity and transmission of Blastocystis infection among the Orang Asli in their communities meaningfully.

  8. 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 

  9. Salleh FM, Moktar N, Yasin AM, Al-Mekhlafi HM, Anuar TS
    J Microbiol Methods, 2014 Nov;106:143-145.
    PMID: 25193442 DOI: 10.1016/j.mimet.2014.08.019
    To improve the stool concentration procedure, we modified different steps of the standard formalin-ether concentration technique and evaluated these modifications by examining stool samples collected in the field. Seven samples were found positive by the modified formalin-ether concentration technique (M-FECT). Therefore, the M-FECT procedure provides enhanced detection of Cryptosporidium oocysts.
  10. Salleh FM, Al-Mekhlafi AM, Nordin A, Yasin 'M, Al-Mekhlafi HM, Moktar N
    Diagn Microbiol Infect Dis, 2011 Jan;69(1):82-5.
    PMID: 21146718 DOI: 10.1016/j.diagmicrobio.2010.08.028
    This study was conducted to evaluate the modification of the usual Gram-chromotrope staining technique developed in-house known as Gram-chromotrope Kinyoun (GCK) in comparison with the Weber Modified Trichrome (WMT) staining technique; as the reference technique. Two hundred and ninety fecal specimens received by the Microbiology Diagnostic Laboratory of Hospital Universiti Kebangsaan Malaysia were examined for the presence of microsporidial spores. The sensitivity and specificity of GCK compared to the reference technique were 98% and 98.3%, respectively. The positive and negative predictive values were 92.5% and 99.6%, respectively. The agreement between the reference technique and the GCK staining technique was statistically significant by Kappa statistics (K = 0.941, P < 0.001). It is concluded that the GCK staining technique has high sensitivity and specificity in the detection of microsporidial spores in fecal specimens. Hence, it is recommended to be used in the diagnosis of intestinal microsporidiosis.
  11. Salam A, Zainuddin Z, Latiff AA, Ng SP, Soelaiman IN, Mohamad N, et al.
    Ann Acad Med Singap, 2008 Sep;37(9):814-6.
    PMID: 18989506
  12. 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.
  13. Anuar TS, Hasim L, Moktar N, Salleh FM, Al-Mekhlafi HM
    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.
  14. Anuar TS, Bakar NH, Al-Mekhlafi HM, Moktar N, Osman E
    PMID: 27405127
    The epidemiology and environmental factors affecting transmission of human microsporidiosis are poorly understood. We conducted the present study to determine the prevalence and risk factors associated with asymptomatic intestinal microsporidiosis among aboriginal school children in the Kuala Krau District, Pahang State, Malaysia. We collected stool samples from 255 school children and examined the samples using Gram-chromotrope Kinyoun stain. We also collected demographic, socioeconomic, environmental and personal hygiene information using a pre-tested questionnaire. Sixty-nine of the children was positive for microsporidia: 72.5% and 27.5% were low (1+) and moderate (2+) excretions of microsporidia spores, respectively. Univariate and multivariate analyses showed being aged 10 years (p = 0.026), using an unsafe water supply as a source for drinking water (p = 0.044) and having close contact with domestic animals (p = 0.031) were all significantly associated with microsporidial infection among study subjects. Our findings suggest asymptomatic intestinal microsporidiosis is common in the study population, more than previously reported. In the study population, control measures need to be implemented, such as good personal hygiene, proper sanitation and safe drinking water supply.
  15. Mohammad NA, Mastuki MF, Al-Mekhlafi HM, Moktar N, Anuar TS
    Iran J Parasitol, 2018;13(1):127-136.
    PMID: 29963095
    BACKGROUND: This study evaluated the performance of routine permanent stain and cultivation method in comparison with polymerase chain reaction assay as the reference technique to detect Blastocystis sp.

    METHODS: A cross-sectional study was conducted among aboriginal populations that reside in Pahang, Peninsular Malaysia in Feb to Mar 2015. A total of 359 stool samples were examined using Wheatley's trichrome stain, in-vitro cultivation in Jones' medium and PCR assay. Positive amplicons were subjected to sequencing and phylogenetic analysis.

    RESULTS: Fifty-six (15.6%) samples were detected positive with Blastocystis sp. by Wheatley's trichrome stain and 73 (20.3%) by in-vitro culture, while PCR assay detected 71 (19.8%) positive samples. Detection rate of Blastocystis sp. was highest in combination of microscopic techniques (27.9%). The sensitivity and specificity of Wheatley's trichrome staining and in-vitro culture techniques compared to PCR assay were 49.3% (95% CI: 37.2-61.4) and 92.7% (95% CI: 89.1-95.4) and 39.4% (95% CI: 28.0-51.8) and 84.4% (95% CI: 79.7-88.4), respectively. However, the sensitivity [60.6% (95% CI: 48.3-71.9)] of the method increased when both microscopic techniques were performed together. False negative results produced by microscopic techniques were associated with subtype 3. The agreement between Wheatley's trichrome stain, in-vitro culture and combination of microscopic techniques with PCR assay were statistically significant by Kappa statistics (Wheatley's trichrome stain: K = 0.456, P<0.001; in-vitro culture: K = 0.236, P<0.001 and combination techniques: K = 0.353, P<0.001).

    CONCLUSION: The combination of microscopic technique is highly recommended to be used as a screening method for the diagnosis of Blastocystis infection either for clinical or epidemiological study to ensure better and accurate diagnosis.

  16. 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.
  17. 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.
  18. 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.
  19. 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.
  20. Ngah NF, Moktar N, Isa NH, Selvara S, Yusof MS, Sani HA, et al.
    Asia Pac J Clin Nutr, 2002;11(2):88-91.
    PMID: 12074186
    This study determined the prevalence of ocular manifestation of vitamin A deficiency in Orang Asli (Aborigine) children. Night blindness was found in 16.0% of the children, conjunctiva xerosis in 57.3%, Bitot's spot in 2.8%, corneal xerosis in 0.5% and corneal scars in 5.6%. These findings show that history of night blindness had sensitivity, specificity and predictive value (positive) of 47.2, 98.1 and 96.2%, respectively, compared with the standard diagnosis procedure using luxometer readings.
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