Molecular detection of porcine Enterocytozoon bieneusi infection in Peninsular Malaysia and epidemiological risk factors associated with potentially zoonotic genotypes.

Enterocytozoon bieneusi is an emerging opportunistic pathogen infecting humans, and both domestic and wild pigs are known to harbour zoonotic genotypes. There remains a paucity of information on the prevalence and epidemiology of this enteropathogen in Southeast Asia. The present study was undertaken to determine the molecular prevalence and risk factors associated with E. bieneusi infection among commercially farmed pigs in Malaysia. Faecal samples were collected from 450 pigs from 15 different farms and subjected to nested PCR amplification of the ribosomal internal transcribed spacer (ITS) gene of E. bieneusi. Phylogenetic analysis involved 28 nucleotide sequences of the ITS region of E. bieneusi. An interviewer-administered questionnaire provided information on the animal hosts, farm management systems and environmental factors and was statistically analysed to determine the risk factors for infection. The prevalence of E. bieneusi infection was relatively high (40.7%). The highest prevalence (51.3%) was recorded among the piglets, while the adults showed the lowest level of infection (31.3%). Multivariate analysis indicated that age of the pigs, distance of the farm from human settlement and farm management system were significant risk factors of infection. Three genotypes (EbpA, EbpC and Henan-III) detected among the pigs are potentially zoonotic. The high prevalence of E. bieneusi among locally reared pigs, the presence of zoonotic genotypes and the spatial distribution of pig farms and human settlements warrant further investigation on the possibility of zoonotic transmission.

Helminths and the microbiota: parts of the hygiene hypothesis.

In modern societies, diseases that are driven by dysregulated immune responses are increasing at an alarming pace, such as inflammatory bowel diseases and diabetes. There is an urgent need to understand these epidemiological trends, which are likely to be driven by the changing environment of the last few decades. There are complex interactions between human genetic factors and this changing environment that is leading to the increasing prevalence of metabolic and inflammatory diseases. Alterations to human gut bacterial communities (the microbiota) and lowered prevalence of helminth infections are potential environmental factors contributing to immune dysregulation. Helminths have co-evolved with the gut microbiota and their mammalian hosts. This three-way interaction is beginning to be characterized, and the knowledge gained may enable the design of new therapeutic strategies to treat metabolic and inflammatory diseases. However, these complex interactions need to be carefully investigated in the context of host genetic backgrounds to identify optimal treatment strategies. The complex nature of these interactions raises the possibility that only with highly personalized treatment, with knowledge of individual genetic and microbiota communities, will therapeutic interventions be successful for a majority of the individuals suffering from these complex diseases of immune dysregulation.

Mitochondrial DNA analyses reveal low genetic diversity in Culex quinquefasciatus from residential areas in Malaysia.

The present study explored the intraspecific genetic diversity, dispersal patterns and phylogeographic relationships of Culex quinquefasciatus Say (Diptera: Culicidae) in Malaysia using reference data available in GenBank in order to reveal this species' phylogenetic relationships. A statistical parsimony network of 70 taxa aligned as 624 characters of the cytochrome c oxidase subunit I (COI) gene and 685 characters of the cytochrome c oxidase subunit II (COII) gene revealed three haplotypes (A1-A3) and four haplotypes (B1-B4), respectively. The concatenated sequences of both COI and COII genes with a total of 1309 characters revealed seven haplotypes (AB1-AB7). Analysis using tcs indicated that haplotype AB1 was the common ancestor and the most widespread haplotype in Malaysia. The genetic distance based on concatenated sequences of both COI and COII genes ranged from 0.00076 to 0.00229. Sequence alignment of Cx. quinquefasciatus from Malaysia and other countries revealed four haplotypes (AA1-AA4) by the COI gene and nine haplotypes (BB1-BB9) by the COII gene. Phylogenetic analyses demonstrated that Malaysian Cx. quinquefasciatus share the same genetic lineage as East African and Asian Cx. quinquefasciatus. This study has inferred the genetic lineages, dispersal patterns and hypothetical ancestral genotypes of Cx. quinquefasciatus.

Spatial density of Aedes distribution in urban areas: a case study of breteau index in Kuala Lumpur, Malaysia.

BACKGROUND & OBJECTIVES: Dengue fever (DF) is a major vector-borne disease in Malaysia. The incidences of DF in Malaysia are caused by viruses transmitted through the bites of infected female Aedes albopictus and Ae. aegypti mosquitoes. This study aims to establish the spatial density of mosquito population or breteau index (BI) in the areas of Kuala Lumpur using geographic information system (GIS), remote sensing (RS) and spatial statistical tools. METHOD: The 2010 weekly report data of BI obtained from the Ministry of Health (MoH) and the 2010 monthly rainfall data obtained from Malaysia Meteorological Services Department were analyzed using RS and spatial statistical tools to show the spatial correlation of dengue in each zone in Kuala Lumpur. The Kernel density was implemented to identify the better dengue hotspot localities. RESULTS: RESULTS indicated a strong significant positive relationship between the number of localities with high BI and monthly rainfall (r = 0.64; p = 0.024). In addition, types of landuse did not appear to influence the mosquito's population (Built-up: r = 0.16, p = 0.118; Cleared area: r = - 0.107, p = 0.304; Vegetation dense: r = - 0.206, p = 0.046; Vegetation sparse: r = 0.023, p = 0.823; and Water: r = 0.246, p = 0.016). INTERPRETATION & CONCLUSION: In the present study, several hotspots identified will be beneficial to assist the local health authorities to reduce and eradicate mosquitoes in these areas. These results will provide valuable information through the application of advanced tools in combating Aedes mosquitoes.

First molecular characterization of Cryptosporidium in Yemen.

Cryptosporidium is a protozoan parasite of humans and animals and has a worldwide distribution. The parasite has a unique epidemiology in Middle Eastern countries where the IId subtype family of Cryptosporidium parvum dominates. However, there has been no information on Cryptosporidium species in Yemen. Thus, this study was conducted in Yemen to examine the distribution of Cryptosporidium species and subtype families. Fecal samples were collected from 335 patients who attended hospitals in Sana'a city. Cryptosporidium species were determined by PCR and sequence analysis of the 18 s rRNA gene. Cryptosporidium parvum and C. hominis subtypes were identified based on sequence analysis of the 60 kDa glycoprotein (gp60) gene. Out of 335 samples, 33 (9.9%) were positive for Cryptosporidium. Of them, 97% were identified as C. parvum whilst 1 case (3%) was caused by C. hominis. All 7 C. parvum isolates subtyped belonged to the IIaA15G2R1 subtype. The common occurrence of the zoonotic IIa subtype family of C. parvum highlights the potential occurrence of zoonotic transmission of cryptosporidiosis in Yemen. However, this postulation needs confirmation with future molecular epidemiological studies of cryptosporidiosis in both humans and animals in Yemen.

Poverty related diseases amongst Malaysia's low-income community: a review.

Poverty, as proven by several studies, is a driving force behind poor health and hygiene practices. This review attempts to outline common communicable and non-communicable diseases that disproportionately affect Malaysia's 2.91 million low-income households. The current study also looks into the government's housing and healthcare programmes for this demographic to improve their health and well-being. The initial examination yielded incredibly little research on this marginalised community, with event reporting typically generalised to the Malaysian community as a whole rather than analysing disease incidences based on household income, which would better reflect povertydriven diseases. As a result, there is an acute need for more accurate information on the epidemiology of diseases among the poor in order to address this public health issue and provide conclusions that can drive policy designs.

Higher efficacy of a single dosage albendazole and different soil-transmitted helminths re-infection profiles amongst indigenous Negritos from inland jungle versus those in resettlement at town peripheries.

Demarginalization through initiation of resettlement program since 1978 is an inevitable progress faced by the indigenous Orang Asli (OA) population in Peninsular Malaysia. As Malaysian huntergatherers, the Negrito has been exposed to various environmental-cultural variations. These changes may influence the pattern of soil-transmitted helminth (STH) infections, the common malady amongst OA. This study evaluated the deworming effects of single-dosage albendazole (400 mg) and STH-reinfection rate between Negritos who are still living in the inland jungle versus those living in resettlements at town peripheries (RPS). Stool samples from the consented participants were first examined using the direct faecal smear, formalin-ether sedimentation and Kato Katz techniques. Subsequently, stool collections were carried out in three time points following treatment (i.e., 21 days, 3 months and 6 months). In brief, a total number of 54 Negritos (inland: 24; RPS: 30) with a complete set of stool collection was included in this longitudinal study. This study revealed 72.2% cure rate against T. trichiura in the inland but only 15.0% in the RPS. Although the efficacy of albendazole against T. trichiura was ultimately low in the RPS, 62.6% egg reduction rate (ERR) (arithmetic mean) was noted (p = 0.001). For A. lumbricoides and hookworm, high cure rates were found in both communities (85.7-100.0%). Reinfection for T. trichiura was seen in less than 1 month with higher rate in the RPS (90.0%) as opposed to the inland (44.4%) at 21 days following treatment. This study found that the inland OA had better tolerability to single-dosage albendazole and experienced slower STH reinfection rates versus the RPS. Hence, the selection of albendazole dosage should be targeted and the use of single- dosage albendazole (biannually) would be more suitable for the inland OA. Conversely, we propose the use of 3-days albendazole regimens in the resettled RPS population.

Imported case ofLeishmania tropica cutaneous leishmaniasis in a 10-year-old child in Malaysia.

The present paper reported a first imported case of cutaneous leishmaniasis in a 10-year- old child who returned from Saudi Arabia to Malaysia. Six weeks after his travel to Malaysia, two erythematous dermal nodules were developed over his right cheek and chin. Occurrence of intracellular amastigote of Leishmania was observed through examination of skin biopsy with hematoxylin and eosin stain. Furthermore, molecular analysis of ribosomal internal transcribed spacer 1 (ITS1) of Leishmania spp. confirmed the child was infected with Leishmania tropica. The child was given oral fluconazole and he had a 80% recovery before he went back to Saudi Arabia.

In silico structural modeling and quality assessment of Plasmodium knowlesi apical membrane antigen 1 using comparative protein models.

Plasmodium knowlesi is the most common zoonotic parasite associated with human malaria infection in Malaysia. Apical membrane antigen 1 (AMA1) protein in the parasite plays a critical role in parasite invasion into host cells. To date, there is no complete three-dimensional ectodomain structure of P. knowlesi AMA1 (PkAMA1) protein. The knowledge of a protein structure is important to understand the protein molecular functions. Three in silico servers with respective structure prediction methods were used in this study, i.e., SWISS-MODEL for homology modeling and Phyre2 for protein threading, which are template-based modeling, while I-TASSER for template-free ab initio modeling. Two query sequences were used in the study, i.e., native ectodomain of PkAMA1 strain H protein designated as PkAMA1-H and a modified PkAMA1 (mPkAMA1) protein sequence in adaptation for Pichia pastoris expression. The quality of each model was assessed by ProSA-web, QMEAN and SAVES v6.0 (ERRAT, Verify3D and Ramachandran plot) servers. Generated models were then superimposed with two models of Plasmodium AMA1 deposited in Protein Data Bank (PDB), i.e., PkAMA1 (4UV6.B) and Plasmodium vivax AMA1 (PvAMA1, 1W81) protein structures for similarity assessment, quantified by root-meansquare deviation (RMSD) value. SWISS-MODEL, Phyre2 and I-TASSER server generated two, one and five models, respectively. All models are of good quality according to ProSA-web assessment. Based on the average values of model quality assessment and superimposition, the models that recorded highest values for most parameters were selected as best predicted models, i.e., model 2 for both PkAMA1-H and mPkAMA1 from SWISS-MODEL as well as model 1 of PkAMA1-H and model 3 of mPkAMA1 from I-TASSER. Template-based method is useful if known template is available, but template-free method is more suitable if there is no known available template. Generated models can be used as guidance in further protein study that requires protein structural data, i.e., protein-protein interaction study.

A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces: development and validation.

Soil-transmitted helminth (STH) infections, mainly caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms, are among the most common intestinal parasites that infect humans. The infections are widely distributed throughout tropical and subtropical countries, including Malaysia, particularly in underprivileged communities. Microscopic and culture techniques have been used as a gold standard for diagnostic techniques. However, these methods yield low sensitivity and specificity, laborious and time-consuming. Therefore, simple, rapid, and accurate alternative methods are needed for the simultaneous detection of STH infections. Although advanced technologies such as real-time multiplex PCR have been established, the use of this technique as a routine diagnostic is limited due to the high cost of the instrument. Therefore, a single-round multiplex conventional PCR assay for rapid detection of four STH species in the fecal sample was developed in this study. To perform the single-round multiplex PCR, each pair of species-specific primers was selected from target genes, including Ancylostoma duodenale (Internal Transcribed Spacer 2; accession No. AJ001594; 156 base pair), Necator americanus (ITS 2; accession No. AJ001599; 225 base pair), Ascaris lumbricoides (Internal Transcribed Spacer 1; accession No. AJ000895; 334 base pair) and Trichuris triciura (partial ITS 1, 5.8s rRNA and partial ITS 2; accession No. AM992981; 518 base pair). The results showed that the newly designed primers could detect the DNA of STH at low concentrations (0.001 ng/ µl) with no cross-amplification with other species. This assay enables the differentiation of single infections as well as mixed infections. It could be used as an alternative and is a convenient method for the detection of STHs, especially for the differentiation of N. americanus and A. duodenale.

Molecular identification of Cryptosporidium parvum from avian hosts.

Cryptosporidium species are protozoan parasites that infect humans and a wide variety of animals. This study was aimed at identifying Cryptosporidium species and genotypes isolated from avian hosts. A total of 90 samples from 37 different species of birds were collected throughout a 3-month period from April 2008 to June 2008 in the National Zoo of Kuala Lumpur, Malaysia. Prior to molecular characterization, all samples were screened for Cryptosporidium using a modified Ziehl-Neelsen staining technique. Subsequently samples were analysed with nested-PCR targeting the partial SSU rRNA gene. Amplicons were sequenced in both directions and used for phylogenetic analysis using Neighbour-Joining and Maximum Parsimony methods. Although 9 (10%) samples were positive for Cryptosporidium via microscopy, 8 (8.9%) produced amplicons using nested PCR. Phylogenetic trees identified all the isolates as Cryptosporidium parvum. Although C. parvum has not been reported to cause infection in birds, and the role of birds in this study was postulated mainly as mechanical transporters, these present findings highlight the significant public health risk posed by birds that harbour the zoonotic species of Cryptosporidium.

Intestinal parasitic infections among children in central Albania.

Although intestinal parasitic infections (IPI) among children remain a global issue, the current information on such infections in Albanian children is very limited. A cross-sectional study of the IPI in 321 children living in the Albanian counties of Tirana (152) and Elbasan (169) was therefore conducted in 2008, with a pre-tested standard questionnaire employed to gather the relevant personal and clinical data. Using formalin-ether concentration and permanent stains, stool samples were examined microscopically for the ova, cysts and oocysts of any parasites. The overall prevalence of IPI was 19% (61 of 321), with protozoan infections (11·5%) apparently more common than infections with soil-transmitted helminths (STH; 8·1%). Giardia duodenalis was the parasite most frequently detected (10·9%), followed by hookworm (5·6%), Ascaris lumbricoides (1·9%), Trichuris trichiura (0·6%), Cryptosporidium (0·3%) and Entamoeba histolytica/dispar (0·3%). The results of a univariate analysis indicated that the children from Tirana county were significantly more likely to be found infected with STH compared with the children from Elbasan county (12·5% v. 4·1%; P=0·006). Children sampled in the community were also more likely to be found STH-positive than the children sampled as they attended hospitals and health clinics (10·5% v. 6·0%) but this difference did not reach statistical significance. The children found STH-positive were five times more likely to be suffering from diarrhoea than the other children checked in clinical settings (P=0·004) and were also more likely to be suffering from abdominal pain (P=0·054) and/or diminished appetite (P=0·016).

Prevalence of bronchial asthma among orang asli in peninsular malaysia.

A survey was carried out to determine the prevalence of bronchial asthma and their contributing risk factors among Orang Asli subgroups living in Malaysia using IUATLD questionnaire and spirometry without being discriminatory towards age or gender. Of the 1171 distributed questionnaires, 716 (61.1%) comprising of 62.7% Semai Pahang, 51.3% Temiar, 74.2% Mah Meri, 65.6% Semai Perak, 53.6% Temuan, 53.8% Semelai, 61.1% Jakun and 67.4% Orang Kuala subgroups completed their questionnaire and were included in the data analysis. Participants comprised 549 (76.7%) children and 167 (23.3%) adults, age between 1 to 83 years old, 304 (42.5%) males and 412 (57.5%) females. The overall prevalence of bronchial asthma was 1.4% of which 1.5% was children, 1.3% adults, 1.0% male and 1.7% female, respectively. Of the 8 subgroups surveyed, 5 out of 10 confirmed asthma cases were Semai Pahang, followed by 3 cases among Mah Meri, and one case each among Temuan and Semai Perak subgroups, respectively. This study also demonstrated that the prevalence of self-reported and confirmed bronchial asthma tend to be higher among those who had close contact with pets, smoking individuals and among those who had a family history of asthma.

Epidemiological study of human intestinal parasites in Sarawak, East Malaysia: A review.

Intestinal parasitic infections are endemic in rural settings and may account for asymptomatic infections to various health complications. These infections are a cause of concern for communities of lower economic status, especially in developing countries. In Sarawak, indigenous populations residing in geographically inaccessible areas are socially and economically disadvantaged. Through close association with nature, these populations are prone to intestinal parasitism. Currently, scattered information has led to a continual state of neglect at each level of parasitic infection control. This urges for a review of their distribution and transmission based on previous reports to understand the pattern of the diseases in the state which can further address the improvement of mass controlling programs. A literature search was conducted to collect previous reports on human intestinal parasites in Sarawak, East Malaysia from PubMed (Medline), SCOPUS, ScienceDirect and Web of Science from January 2019 to March 2021. Extrapolating the current data in Sarawak which is still considered limited, further interdisciplinary strategies are demanded to give insights in the epidemiology and true prevalence of intestinal parasites in Sarawak. This review addresses for redirection of attitude towards intestinal parasitic infections where it should be given with ample attention by rural populations. In tandem to that, improvement of rural livelihood such as standard of living and sanitation in Sarawak should be accredited as part of the efforts to reduce the number of intestinal parasitic infections in the state. As a control measure, mass deworming should be reconsidered especially to the rural populations.

Serological survey of canine vector-borne diseases in two animal shelters in central Peninsular Malaysia.

Canine vector-borne diseases (CVBDs) are increasingly becoming a cause for global concern because of their high morbidity and mortality rates in dogs. However, information on their occurrence in Malaysia is still scanty. In this study, a total of 103 dog blood samples were collected from two animal shelters in central Peninsular Malaysia and tested for the antibodies against Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and the antigen of Dirofilaria immitis. Of the 103 tested dogs, 44.7% (46) were found to be seropositive for Ehrlichia spp., 30.1% (31) for Anaplasma spp. and 13.6% (14) for D. immitis. Co-infections of Anaplasma spp. + Ehrlichia spp. (18.5%, 19) were most prevalent, followed by Anaplasma spp. + D. immitis (1.9%; two) and D. immitis + Ehrlichia spp. (1.0%; one). Furthermore, three dogs (2.9%) were also found to have triple infection, testing seropositive for Ehrlichia spp., Anaplasma spp. and D. immitis. The dogs which were found to be seropositive with at least one pathogen were 66.7% (32/51) at shelter A, and 55.8% (29/52) at shelter B. Serological evidence showed that the exposure of major vector-borne diseases in dogs in shelters was relatively high in the surveyed areas. Routine detection and control of vector-borne diseases are of paramount importance for reducing the risk of CVBDs transmission in dogs and humans.

Bioinformatics characterization of Plasmodium knowlesi apical membrane antigen 1 (PkAMA1) for multi-epitope vaccine design.

Malaria caused by Plasmodium knowlesi species has become a public health concern, especially in Malaysia. Plasmodium knowlesi parasite which originates from the macaque species, infects human through the bite of the Anopheles mosquitoes. Research on malaria vaccine has been a continuous effort to eradicate the malaria infection, yet there is no vaccine against P. knowlesi malaria to date. Apical membrane antigen 1 (AMA1) is a unique surface protein of all apicomplexan parasites that plays a crucial role in parasite-host cell invasion and thus has been a long-standing malaria vaccine candidate. The selection of protective epitopes in silico has led to significant advances in the design of the vaccine. The present study aimed to employ bioinformatics tools to predict the potential immunogenic B- and T-cell epitopes in designing malaria vaccine targeting P. knowlesi AMA1 (PkAMA1). B-cell epitopes were predicted using four bioinformatics tools, i.e., BepiPred, ABCpred, BcePred, and IEDB servers whereas T-cell epitopes were predicted using two bioinformatics servers, i.e., NetMHCpan4.1 and NetMHCIIpan-4.0 targeting human major histocompatibility complex (MHC) class I and class II molecules, respectively. The antigenicity of the selected epitopes computed by both B- and T-cell predictors were further analyzed using the VaxiJen server. The results demonstrated that PkAMA1 protein encompasses multi antigenic regions that have the potential for the development of multi-epitope vaccine. Two B- and T-cell epitopes consensus regions, i.e., NSGIRIDLGEDAEVGNSKYRIPAGKCP (codons 28-54) and KTHAASFVIAEDQNTSY RHPAVYDEKNKT (codons 122-150) at domain I (DI) of PkAMA1 were reported. Advancement of bioinformatics in characterization of the target protein may facilitate vaccine development especially in vaccine design which is costly and cumbersome process. Thus, comprehensive B-cell and T-cell epitope prediction of PkAMA1 offers a promising pipeline for the development and design of multi-epitope vaccine against P. knowlesi.

Bioefficacy of mosquito mat vaporizers and associated metabolic detoxication mechanisms in Aedes aegypti (Linnaeus) in Selangor, Malaysia: A statewide assessment.

This study aims to examine the efficacy of mosquito mat vaporizers on Aedes aegypti and their associated metabolic detoxication mechanisms. For this purpose, Aedes aegypti (Linnaeus) was collected from nine districts in Selangor, Malaysia and tested with mosquito vaporizing mat bioassays. The same populations were also subjected to biochemical assays to investigate activities of detoxifying enzymes, namely non-specific esterase (EST), glutathione-S-transferase (GST) and mixed function oxidase (MFO). The efficacy of Ae. aegypti on the active ingredients tested in decreasing order were d- allethrin > dimefluthrin > prallethrin with PBO > prallethrin. The results further indicated significant enhancement mean levels of EST, GST and MFO in pyrethroid-resistant populations. The mortality rate of Ae. aegypti in response to pyrethroid active ingredients was associated with MFO activity, suggesting it is an important detoxification enzyme for the populations tested. In view of the presence of resistance against household insecticide products, pyrethroid efficacy on Ae. aegypti populations needs to be monitored closely to ensure the implementation of an effective vector control program in Malaysia.

Oral bacteria detection among children with cancer in a tertiary teaching hospital in Kuala Lumpur, Malaysia.

This study sought to determine the prevalence of pathogenic and non-pathogenic bacteria in the oral cavities of children with cancer. There were 68 paediatric patients with cancer who were included in this study. Oral swab samples from the dorsum of tongues and mouth floors of these patients were subjected to culture, staining, and molecular methods to detect the bacteria. The overall prevalence of gram-positive and gram-negative bacteria was 79.4% (54/68; 95% CI = 68.4 - 87.3) and 25% (17/68; 95% CI = 16.2 - 36.4), respectively. Streptococcus salivarius and Streptococcus parasanguinis were the predominant pathogenic grampositive bacteria, while Neisseria subflava and Neisseria perflava were the most common pathogenic gram-negative bacteria. The results revealed that the number of bacteria isolates recovered in patients receiving cancer treatment was higher (55.9%) than those who had not received treatment (16.2%). Therefore, more isolated pathogenic bacteria were observed post-therapy (54.4%). Pathogenic organisms can have significant implications on patient health. Awareness of the types of bacteria inhabiting the oral cavity is essential to predict and prevent dental problems, and their associated systemic complications. Findings on the diversity of oral microflora can also provide a better understanding of the aetiology of oral diseases in paediatric patients receiving cancer treatment.

Toxoplasma gondii infection among selected indigenous community in Sarawak, East Malaysia.

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis in humans. To date, little is known about T. gondii infection among the indigenous community, particularly in East Malaysia. This study was conducted to determine the status of T. gondii infection and to investigate associated risk factors among the indigenous community of Sarawak, East Malaysia. The sociodemographic data was obtained using a pretested questionnaire. A serological test was done to detect the presence of specific IgM and IgG antibodies against T. gondii in serum samples. A nested polymerase chain reaction (PCR) was used to determine acute infection among seropositive individuals. The overall seroprevalence of T. gondii infection was 50% (95% CI = 43.3 - 56.7). From this subset, 40.1%, 5.7%, and 4.2% were positive for anti-T. Gondii IgG antibodies, IgM, and both IgG and IgM, respectively. Four seropositive samples were amplified through PCR. None of the pregnant women tested positive for T. gondii infection based on the serological and PCR assays. A significant association was found between age, low monthly household income, unemployment, usage of untreated water and close contact with T. gondii seropositive cats. These results provide basic information on T. gondii infection and may be useful for policymakers to initiate prevention and control programs, especially amongst pregnant women and women of childbearing age in the indigenous community.

Molecular characterization of Giardia duodenalis isolated from Semai Pahang Orang Asli (Peninsular Malaysia aborigines).

This study was conducted to determine the genotypes of Giardia duodenalis isolated from human faecal samples at Pos Betau, Pahang, Malaysia. Faecal specimens were collected and examined for G. duodenalis cysts using Trichrome staining techniques. Molecular identification was carried out by the amplification of a region of the small subunit of the nuclear ribosomal RNA (SSU rRNA) gene using nested PCR and subsequent sequencing. The sequences from 15 isolates from G. duodenalis were subjected to phylogenetic analysis (including appropriate outgroups) using the neighbor-joining and maximum parsimony methods. The trees identified G. duodenalis assemblages A and B, with a predominance of assemblage B. The predominance of anthroponotic genotypes indicates the possibility of anthroponotic transmission of these protozoa in this Semai Pahang Orang Asli community.