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Establishing a protocol for water sample processing for the detection of Blastocystis sp

Lee IL, Tan TC, Govind SK

Exp Parasitol, 2019 Mar;198:105-110.
PMID: 30695704 DOI: 10.1016/j.exppara.2019.01.007

This study was aimed at establishing a protocol for water sample processing for the detection of Blastocystis sp. using distilled water spiked with Blastocystis sp. cysts. The study established a protocol involving eight technical aspects, namely, storage temperature, storage duration, minimum water sample volume, optimum relative centrifugal force, centrifugation duration, minimum number of cyst for inoculation in Jones' medium and turn-around-time for the detection of vacuolar forms of Blastocystis sp. Results showed a minimum of 1.0 L water sample should be collected and processed on the same day. Otherwise, it should be stored at 4 °C and processed within 3 days. Water sample should be centrifuged at 1400×g for 10 min. For the isolation of Blastocystis sp. cysts, parasite pellet could be layered on top of Ficoll-Paque™ PLUS, centrifuged at 1400×g for 20 min and washed twice using 0.9% saline with centrifugation at 1400×g for 10 min. A minimum of 1 × 105 cysts could then be inoculated in Jones' medium supplement with 10% horse serum, incubated at 37 °C and examined for any presence of vacuolar forms of Blastocystis sp. after 3 days of inoculation. A protocol for water sample processing for the detection of Blastocystis sp. has successfully been established. The protocol was validated using 106 various water samples. This protocol will be very useful in determining the extent of Blastocystis sp. contamination in water sources in order to identify the seriousness of contamination.
Single dose treatment with Tiberal of Giardia lamblia infection in children

Iyngkaran N, Lee IL, Robinson MJ

Scand. J. Infect. Dis., 1978;10(3):243-6.
PMID: 362519

A new metronidazole derivative, Tiberal (Ro-07-0207, Roche Laboratories), was evaluated in 22 children with Giardia lamblia infection. Seven patients received an oral dose of 1 g twice daily for one day; the remaining 15 patients received a single dose of 50 mg/kg. Parasitological cure was noted in all 22 patients. Significant side effects were observed only in those children who received the drug at the higher dosage regime. The present study also confirms the findings of other authors that a mucosal imprint method is more reliable than examination of stools, duodenal juice or jejunal biopsy material for the detection of G. lamblia infection.
Fulltext Role of Seasonal Variation on The Prevalence and Risk Factors of Trichuriasis Among the Aboriginal Community in Malaysia

Noradilah SA, Anuar TS, Lee IL

Helminthologia, 2022 Mar;59(1):55-63.
PMID: 35601764 DOI: 10.2478/helm-2022-0004

Trichuris trichiura is a soil-transmitted helminth prevalent in developing countries with poor, inadequate sanitation and unsafe water sources. In Malaysia, the prevalence of trichuriasis is relatively high among the aboriginal community due to poverty and poor sanitation. However, there are few studies to determine the seasonal variation on the prevalence and risk factors to acquire Trichuris trichiura infection in Malaysia. The present study found higher Trichuris trichiura infection during the dry season (63.6%; 138/217) in comparison to the wet season (55.5%; 142/256). Low household income, low education level and practice of open defecation were significant risk factors to acquire Trichuris trichiura infection during the wet season. Usage of untreated water supplies for daily activities was a risk factor to acquire trichuriasis during the dry season. This study highlighted that poverty and poor sanitation practices as well as drinking untreated water put the aborigines at high risk to acquire trichuriasis. Therefore, health education, improved sanitation and provision of treated water supply are recommended for the prevention and control of Trichuris trichiura infections in the aboriginal community.
Fulltext The Coexistence of Blastocystis spp. in Humans, Animals and Environmental Sources from 2010-2021 in Asia

Rauff-Adedotun AA, Meor Termizi FH, Shaari N, Lee IL

Biology (Basel), 2021 Sep 30;10(10).
PMID: 34681089 DOI: 10.3390/biology10100990

Blastocystis spp. are controversial unicellular protists that inhabit the gastrointestinal tract of humans and a wide range of animals worldwide. This review provides an overview of the prevalence and distribution of Blastocystis spp. and their subtypes throughout Asia. Research articles reporting on the presence of Blastocystis spp. in locations within Asia, between 1 January 2010, and 10 May 2021, were obtained from Scopus, PubMed, and Google Scholar. In 427 articles, the prevalence of Blastocystis spp. in 31 countries within the last decade was revealed. Isolates were found in humans, various mammals, birds, reptiles, insects, water sources, vegetables, and ambient air. Prevalence of Blastocystis spp. varied widely across host categories. Subtypes identified throughout Asia were STs 1-14, and ST18-22 (novel subtypes). ST1, ST2, ST3, ST4 were the most frequently isolated in humans; ST5 in pigs; ST10 and ST14 in goats, sheep, and cattle; and ST6 and ST7 in chickens. ST1 and ST3 were most common in water samples. ST1, ST2, ST3, ST4, ST5 and ST6 were shared by humans, animals, and water sources. There is a growing interest in the study of Blastocystis spp. and their subtypes in Asia. Due to the isolation of Blastocystis spp. from biotic and abiotic sources in Asia, the application of the One Health (OH) approach to the study of Blastocystis spp. is proposed for improved perception of this organism.
Predominance of Blastocystis sp. subtype 4 in rural communities, Nepal

Lee IL, Tan TC, Tan PC, Nanthiney DR, Biraj MK, Surendra KM, et al.

Parasitol Res, 2012 Apr;110(4):1553-62.
PMID: 22076050 DOI: 10.1007/s00436-011-2665-0

Blastocystis sp. is a common intestinal parasite. To date, there have been sporadic and scanty studies on Blastocystis sp. carried out in rural communities in Nepal. We surveyed the prevalence of Blastocystis sp. and its possible associated risk factors, and reported the predominant Blastocystis sp. subtype in two rural communities, Bolde Phediche and Bahunipati, in Nepal. Human faecal samples were collected from 241 participants, cultured using in vitro cultivation and examined for Blastocystis sp. The presence of Blastocystis sp. in faecal samples was further confirmed by polymerase chain reaction (PCR) and subsequently genotyped using subtype-specific sequence tagged site (STS) primers. There were 26.1% (63/241) of the participants that were infected by Blastocystis sp. We detected 84.1% (53/63) of Blastocystis sp. subtype 4 infections in these rural communities. The unusually high prevalence of Blastocystis sp. subtype 4 can be attributed to the rearing of family-owned animals in barns built close to their houses. Eighty one percent (51/63) of the Blastocystis sp. infected participants drank not boiled or unfiltered water. The present study revealed that Blastocystis sp. could pose a health concern to the communities and travellers to the hilly area in Nepal. Infection may be transmitted through human-to-human, zoonotic and waterborne transmissions. We provide recommendations to ensure good public health practices.
Prevalence, potential risk factors and genetic diversity of Blastocystis in ruminant livestock animals from Penang, Malaysia

Rauff-Adedotun AA, Lee IL, Abd Talib N, Shaari N, Yahaya ZS, Meor Termizi FH

Parasitol Res, 2023 Sep;122(9):2193-2205.
PMID: 37462745 DOI: 10.1007/s00436-023-07920-5

Blastocystis is a unicellular, anaerobic protist inhabiting the intestinal tract of diverse animal hosts, including human. Information regarding Blastocystis in small ruminants, namely goats and sheep, is limited globally; thus, this study was carried out to investigate the distribution and determinants of Blastocystis in ruminant livestock animals from Penang, Malaysia. Fecal samples from 127 cattle, 149 goats, and 100 sheep were examined for Blastocystis by in vitro cultivation using modified Jones' medium, while DNA barcoding was used for subtyping. Overall, 23.1% (87/376) of animals screened were positive for Blastocystis sp. The prevalence of infection was significantly higher in goats than in cattle and sheep, while the female gender, semi-intensive farming system, and the Northeast Penang Island district were identified as potential risk factors for Blastocystis infection. Blastocystis sp. ST5, ST14, and ST25 were identified in cattle; ST5, ST10, ST13, and ST14 in goats; and ST4, ST5, ST14, and ST15 in sheep. ST5 and ST14 were found to be the most abundant and widespread subtypes in the study area. To the best of our knowledge, this is the first report of ST4 from sheep and ST13 from goats, thus serving as an update to the host range of Blastocystis sp. ST4 and ST13. The isolation of ST4 and ST5 in this study suggests that ruminant livestock animals could serve as reservoirs of human infection.
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 Gut microbiome of helminth-infected indigenous Malaysians is context dependent

Tee MZ, Er YX, Easton AV, Yap NJ, Lee IL, Devlin J, et al.

Microbiome, 2022 Dec 07;10(1):214.
PMID: 36476263 DOI: 10.1186/s40168-022-01385-x

BACKGROUND: While microbiomes in industrialized societies are well characterized, indigenous populations with traditional lifestyles have microbiomes that are more akin to those of ancient humans. However, metagenomic data in these populations remains scarce, and the association with soil-transmitted helminth infection status is unclear. Here, we sequenced 650 metagenomes of indigenous Malaysians from five villages with different prevalence of helminth infections.
RESULTS: Individuals from villages with higher prevalences of helminth infections have more unmapped reads and greater microbial diversity. Microbial community diversity and composition were most strongly associated with different villages and the effects of helminth infection status on the microbiome varies by village. Longitudinal changes in the microbiome in response to albendazole anthelmintic treatment were observed in both helminth infected and uninfected individuals. Inference of bacterial population replication rates from origin of replication analysis identified specific replicating taxa associated with helminth infection.
CONCLUSIONS: Our results indicate that helminth effects on the microbiota were highly dependent on context, and effects of albendazole on the microbiota can be confounding for the interpretation of deworming studies. Furthermore, a substantial quantity of the microbiome remains unannotated, and this large dataset from an indigenous population associated with helminth infections is a valuable resource for future studies. Video Abstract.