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. � 2017, Malaysian Society for Parasitology. All rights reserved.

Acanthamoeba is a free-living protozoa which causes serious ocular problem. Acanthamoeba keratitis is becoming more prevalent amongst contact lens wearers and it can cause loss of vision and blindness if not treated properly. The objective of this research is to determine the effectiveness of gentamicin against six Acanthamoeba spp. isolates, of which three were clinical isolates (HS 6, HKL 95, HTH 73) and three environmental isolates (SMAL 7, SMAL 8, TTT 9). Cyst suspension from the chosen isolates were exposed to gentamicin. After 48 hours of incubation at temperature of 30�C and 37�C, each mixture was filtered and filtration membrane was put onto non-nutrient agar laid with Escherichia coli. The agar plates were incubated for three days at 30oC and 37oC and the plates were examined daily until day 14 to look for the presence of Acanthamoeba trophozoites under inverted microscope. The presence of trophozoites indicated the ineffectiveness of gentamicin. Gentamicin was found to be effective against Acanthamoeba cysts from all the test strains at both incubation temperatures. The minimum cysticidal concentration (MCC) mean value of gentamicin was 0.193 mg/mL at 30oC and 0.229 mg/mL at 37oC. So, we concluded that gentamicin has cysticidal potential towards Acanthamoeba.

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 from 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.

Microscopy technique is widely used in the detection of Blastocystis sp. infection. This study was performed to compare two microscopy-based techniques; in vitro cultivation of faecal specimens in Jones' medium (IVC) followed by Wheatley Trichrome staining and fixed-PVA faecal specimens (PVA) followed by Wheatley Trichrome staining with single-round polymerase chain reaction (PCR) as the reference technique. Examination was performed on 466 faecal samples obtained from Orang Asli community in Pahang, Malaysia. Detection rate of Blastocystis sp. was highest in IVC (35.6%) followed by PVA (20.0%). Single-round PCR detected 41.0% of the faecal specimens. The sensitivity and specificity of PVA and IVC compared to the reference technique were 75.3% (95% CI: 65.2-83.6) and 68.5% (CI: 63.7-73.3), and 88.6% (CI: 82.7-93.0) and 86.3% (CI: 81.9-90.0) respectively. The agreement between the reference technique and PVA showed fair statistically significant agreement by Cohen Kappa statistics of (K=0.318, p<0.001), meanwhile substantial statistically significant agreement was observed between PCR and IVC by Cohen Kappa (K=0.727, p<0.001). Therefore, in vitro cultivation in Jones' medium followed by Wheatley Trichrome staining of faecal specimens should be used as a screening technique in the detection Blastocystis sp. infections. � 2017 Oriental Scientific Publishing Company. Al rights reserved.

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 < 0.05). Analysis of the association of each Blastocystis subtype with socioeconomic characteristics showed the presence of other family members infected with Blastocystis ST3 and the use of stored river water for domestic activities were the significant risk factors for Blastocystis ST3 infections during both seasons. Untreated water supply and low monthly household income (less or equal to RM 500) were the other significant risk factors for Blastocystis ST3 infections during wet and dry season, respectively. The presence of other family members with Blastocystis ST1 and ST2 was the only significant risk factor associated with ST1 and ST2 infections during both seasons. We hypothesise that transmission of Blastocystis ST1, ST2 and ST3 occurred from person to person during both seasons. The waterborne transmission was also identified as a mode of transmission of Blastocystis ST3. Conclusion: The significant risk factors identified in this study were important in the dynamic transmission of Blastocystis infections during both seasons. Provision of treated water supply and health education are affirmative actions to be taken to control Blastocystis infections in this community. 2017 The Author(s).

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. � 2016 Noradilah et al.

Introduction: Acanthamoeba is an ubiquitous free-living protozoa which causes serious ocular problems. Acanthamoeba keratitis is becoming more prevalent amongst contact lens wearers. The disease can cause loss of vision and blindness if not treated properly. The objective of this research is to study the sensitivity of six Acanthamoeba spp. isolates, of which three were from the clinical isolates (HKL 95, HTH 40 and HS 6) and the remaining three from environmental isolates (TTT 9, TL 3 and SMAL 8) to antimicrobial agents. Methods: The antimicrobial agents chosen for this purpose were polyhexamethylene biguanide (PHMB) and Chlorhexidine. Serial dilutions were perfomed forpolyhexamemylene biguanide and Chlorhexidine. Cyst suspensions from the chosen isolates were exposed to PHMB and Chlorhexidine respectively. After 48 hours incubation time at 30�C, each mixture was filtered and filtration membrane was put onto non-nutrient agar laid with Escherichia coli. The agar plates were incubated for three days at 30�C and examined daily until day 14 to detect the presence of Acanthamoeba trophozoites under the inverted microscope. The presence of trophozoites indicated me ineffectiveness of the antimicrobial agents. Results: Both of the antimicrobial agents tested were found to be effective against Acanthamoeba cysts from all die test strains. Polyhexamethylene biguanide gave a minimum cysticidal concentration (MCC) mean value of 2.848 ug/mL while Chlorhexidine showed MCC mean value at a concentration of 3.988 ?g/mL. Contusion: It can be concluded that me Acanthamoeba cysts were sensitive to polyhexamethylene biguanide and Chlorhexidine.