Blastocystis has been increasingly reported in water bodies. However, lack of studies to determine the presence of Blastocystis in water used by the aborigines in Malaysia has led to the birth of this research. This study was therefore aimed to determine the occurrence of Blastocystis in water samples in aboriginal settlements in Pahang, Malaysia. Water samples collected from seven sampling points of two rivers and other water sources in the villages were subjected to filtration and cultivation followed by trichrome staining. The trichrome stained slides were observed microscopically under 1000X magnification for the presence of Blastocystis. River samples were also measured for physicochemical parameters. From this study, 42.9% of the river water and 6.25% of other water samples were positive for Blastocystis. All river samples showed presence of Escherichia coli and Enterobacter aerogenes, indicating faecal contamination. Statistical analysis showed Blastocystis occurrence in the river were significantly correlated conductivity, turbidity, chemical oxygen demand (COD), total dissolved solid (TDS), concentration of sulfate and faecal coliforms. The river water used by the aborigines is a probable source for Blastocystis transmission in this community. Therefore, protection of the river from organic material and faecal contaminations are highly required in order to control the contamination by Blastocystis.

Microscopy-based technique has been widely used in the detection of Blastocystis sp. This study was conducted to compare the techniques used for screening of Blastocystis sp., namely in vitro cultivation of stool specimens in Jones' medium (rvc) followed by Wheatley Trichrome staining and direct examination of stool samples preserved with polyvinyl alcohol (PvA) followed by Wheatley Trichrome staining with single-round polymerase chain reaction (PcR) as the reference technique. The study was performed on 466 stool samples obtained from the aboriginal community in Pahang, Malaysia. rvc showed higher detection rate of Blastocystis sp. (35.6%) than PVA (20.0%). Single-round PCR detected Blastocystis sp. in 41.0% of the stool specimens. The sensitivity and specificity of PVA and rvc in comparison to the reference technique were 75.3% (95% ct: 65 2-83 .6) and 68.5% (CI: 63 .7-73 3) and 88.6% (ct: 82.7-93.0) and 863% (CI: 81.9-90.0), respectively. The agreement between the reference technique and PVA showed statistically significant fair agreement by Cohen Kappa statistics of (K=0318, p<0.001), meanwhile statistically significant substantial agreement was observed between PCR and tvc by Cohen Kappa (K=0.727, p<0.001). Therefore, in vitro cultivation in Jones' medium followed by Wheatley Trichrome staining of stool specimens should be used as a screening technique in the detection of Blastocystis sp. infections.

Objective: To determine the prevalence and risk factors of Giardia, (G.) lamblia, infections among the aboriginal community during the wet and dry seasons. Methods: A total of 473 stool samples flout the aborigines in Temerloh, Pahang, Malaysia were collected during wet (n=256) and dry seasons (n=217). Smear of all the PVA-preserved stool samples were subjected to Trichrome staining and microscopic examination under 1 000 x magnification (Nikon eclipse E100) for the detection of G. lamblia. Positivity was recorded based on the presence of G. lamblia in trophozoite and/or cyst forms. Results: The prevalence of giardiasis was 12.10% and 8.29% during the wet and dry season, respectively. Age of less or equal to 15 years old and presence of other family members with G. lamblia infection were found to be the significant risk factors to acquire G. lamblia infections during both seasons. Untreated water supply was the significant risk factor of giardiasis during the dry season. This study highlighted the possibility of anthroponotic transmission of G. lamblia during both seasons and waterborne transmission during the dry season in the aboriginal community. Conclusions: This study suggests that seasonal variation plays an important role in the prevalence and risk factor of G. lamblia, infection in the aboriginal community. Therefore, close contact with Giardia-infected family members and water-related activities or usage of untreated water must be avoided to reduce the burden of G. lamblia infection in this community.

In endemic areas of intestinal parasitic infections, prevalence of Blastocystis sp in animals has not been clearly elucidated. This is the first study of the distribution of Blastocystis sp subtypes in animals reared by Orang Ash population in Pahang, Malaysia during a wet and dry season. Fecal samples of dogs, chickens, goats, ducks, swans, birds and cows were collected and subjected to PCR amplification and sequencing of Blastocystis small subunit rDNA. Of 127 fecal samples collected during the wet season, 9% were positive for Blastocystis sp, with Blastocystis sp ST3 being predominant (16%) followed by ST1 (4%), ST7 (3%), ST4 (2%), ST10 (2%), ST6 (1%), and ST9 (1%). Of 146 fecal samples collected during the dry season 37% were positive, with Blastocystis sp ST3 being predominant (10%) followed by ST1 (8%), ST7 (6%), ST4 (5%), ST8 (3%), ST2 (1%), ST6 (1%), ST9 (1%), and ST10 (1%). High prevalence of Blastocystis sp was observed in dogs and chickens which carried a diverse range of subtypes especially during the dry season. Dogs and chickens might comprise a part of the transmission dynamics of the infection in the population. Health education related to awareness of hygienic practice and disposal of animals waste should be regularly provided and monitored to prevent the transmission of Blastocystis sp infection in this population.

In the tropics, there are too few studies on isolation of Biastocystis 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 stud)/ 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 ()the: 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 resistant subtype able to survive in any adverse environmental condition. robust and Restriction and control of human and animal faecal contaminations to the river and other water