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Blastocystis is a stramenopile protist of controversial pathogenicity. The organism colonizes a broad range of vertebrate and invertebrate hosts. Blastocystis has also been found in the environment both in water and soil. Several routes of transmission have been proposed including human to human, animal to human, and via contaminated food and water. In recent years, the presence of Blastocystis in vegetables has started to be explored. However, most studies have focused on microscopic detection. Moreover, works of this type from Asia are barely available. Hence, the aim of this preliminary study was to examine the occurrence of Blastocystis in raw vegetables sold in markets in northern Thailand. Fresh produce (n = 20) commonly used in Thai cuisine (Lanna) was purchased from two street markets and screened for Blastocystis using qPCR. Blastocystis was detected in 45% of the samples with the dominant subtype being ST3. Produce growing underground, such as galangal, carrot, and beetroot, were positive for the organism suggesting soil or inadequately composted manure as the source of contamination. To our knowledge, our study is the first to perform subtyping of Blastocystis in vegetables. Our results hint toward fresh produce being a, as yet, not widely explored, transmission route of Blastocystis in the studied community. Looking forward, large-scale investigations on the prevalence of this and other organisms under the One Health umbrella should be undertaken. [ABSTRACT FROM AUTHOR]

Blastocystis is the most commonly found eukaryote in the gut of humans and other animals. This protist is extremely heterogeneous genetically and is classified into 28 subtypes (STs) based on the small subunit ribosomal RNA (SSU rRNA) gene. Numerous studies exist on prevalence of the organism, which usually focus on either humans or animals or the environment, while only a handful investigates all three sources simultaneously. Consequently, understanding of Blastocystis transmission dynamics remains inadequate. Our aim was to explore Blastocystis under the One Health perspective using a rural community in northern Thailand as our study area. We surveyed human, other animal and environmental samples using both morphological and molecular approaches. Prevalence rates of Blastocystis were 73% in human hosts (n = 45), 100% in non-human hosts (n = 44) and 91% in environmental samples (n = 35). Overall, ten subtypes were identified (ST1, ST2, ST3, ST4 ST5, ST6, ST7, ST10, ST23, and ST26), eight of which were detected in humans (ST1, ST2, ST3, ST4, ST5, ST7, ST10, and ST23), three in other animals (ST6, ST7, and ST23), while seven (ST1, ST3, ST6, ST7, ST10, ST23, and ST26) were found in the environment. In our investigation of transmission dynamics, we assessed various groupings both at the household and community level. Given the overall high prevalence rate, transmission amongst humans and between animals and humans are not as frequent as expected with only two subtypes being shared. This raises questions on the role of the environment on transmission of Blastocystis. Water and soil comprise the main reservoirs of the various subtypes in this community. Five subtypes are shared between humans and the environment, while three overlap between the latter and animal hosts. We propose soil as a novel route of transmission, which should be considered in future investigations. This study provides a thorough One Health perspective on Blastocystis. Using this type of approach advances our understanding on occurrence, diversity, ecology and transmission dynamics of this poorly understood, yet frequent gut resident. [ABSTRACT FROM AUTHOR]

Blastocystis inhabits the digestive tracts of a diverse range of hosts. Transmission patterns, including host specificity, and the clinical and public health significance of Blastocystis in humans remain poorly understood. This study aimed to investigate the distribution and genetic diversity of Blastocystis in herbivorous and carnivorous reptiles in Eastern Thailand. A total of 501 faecal samples were collected from 363 iguanas, 79 bearded dragons, 50 tortoises, and nine snakes in an animal breeding farm in Chonburi Province, Eastern Thailand. Detection and differentiation of Blastocystis was based on amplification, sequencing, and phylogenetic analysis of specific small subunit (SSU) ribosomal RNA genes from faecal DNA extracted from the samples. Altogether 101/501 samples (20 %) were polymerase chain reaction (PCR) and sequencing-positive for Blastocystis , 90 (89 %) of which were from iguanas; the remaining positive samples were from African spurred tortoise (n=6), Bearded dragon (n=3), Leopard tortoise (n=1), and Red-footed tortoise (n=1). Phylogenetic analysis revealed that most of the Blastocystis sequences from iguanas were largely similar, and they were distinct from those of the tortoises. Subtype 17 was found in the three bearded dragons and likely reflected Blastocystis from prey animals. This is the largest survey of Blastocystis in reptiles to date. Remarkable differences in Blastocystis colonization rates and genetic diversity were observed between iguanas and other reptile orders, and what was considered Blastocystis colonization was only observed in herbivorous reptiles. • Blastocystis was commonly found in herbivores. • ST17 as an incidental finding. • This study is possibly the largest survey of Blastocystis in reptiles. [ABSTRACT FROM AUTHOR]

Abstract Blastocystis is a unicellular protist most commonly detected in humans and a variety of animals. The predominant mode of its transmission is the fecal–oral route, but its zoonotic potential is not completely understood. The objective of this study was to determine the presence and genetic diversity of Blastocystis on pig farms in Nakhon Pathom Province, Central Thailand. A total of 154 human and 90 pig stool samples were collected and analyzed. Nested PCR detected Blastocystis in 35.55% of the pig samples and 6.49% of the human samples. Subtyping based on regions of the small-subunit ribosomal RNA (SSU rRNA) gene identified three Blastocystis subtypes in pigs and humans: ST1, ST3, and ST5. Blastocystis ST5 was the predominant subtype, followed by ST1 and then ST3. All the sequences from the Blastocystis -positive samples from both pigs and humans were closely related. This study reveals a possibility of low host specificity of Blastocystis STs (ST1, ST3 and ST5) on pig farms in Thailand. We tentatively suggest that close contact with or exposure to pig stools may be a significant source of Blastocystis detected in pig handlers. Further studies are required to confirm the zoonotic transmission of this organism in Thailand, because pigs may play an important role in the transmission of Blastocystis. Graphical abstract Unlabelled Image Highlights • High percentages of Blastocystis infection in pigs • Blastocystis ST5 was the predominant subtype in both humans and pigs. • A possibility of low host specificity of Blastocystis STs (ST1, ST3 and ST5) on pig farms in Thailand [ABSTRACT FROM AUTHOR]

Abstract Blastocystis is a common and broadly distributed microbial eukaryote inhabiting the gut of humans and other animals. The genetic diversity of Blastocystis is extremely high comprising no less than 17 subtypes in mammals and birds. Nonetheless, little is known about the prevalence and distribution of Blastocystis subtypes colonising humans in Thailand. Molecular surveys of Blastocystis remain extremely limited and usually focus on the central, urban part of the country. To address this knowledge gap, we collected stool samples from a population of Thai adults (n = 178) residing in Chiang Rai Province. The barcoding region of the small subunit ribosomal RNA was employed to screen for Blastocystis and identify the subtype. Forty-one stool samples (23%) were identified as Blastocystis positive. Six of the nine subtypes that colonise humans were detected with subtype (ST) three being the most common (68%), followed by ST1 (17%) and ST7 (7%). Comparison of subtype prevalence across Thailand using all publicly available sequences showed that subtype distribution differs among geographic regions in the country. ST1 was most commonly encountered in the central region of Thailand, while ST3 dominated in the more rural north and northeast regions. ST2 was absent in the northeast, while ST7 was not found in the center. Thus, this study shows that ST prevalence and distribution differs not only among countries, but also among geographic regions within a country. Potential explanations for these observations are discussed herewith. Graphical abstract Unlabelled Image Highlights • First study of Blastocystis prevalence and subtyping in Chiang Rai Province, Thailand. • Six of the nine subtypes known to colonise humans were found. • ST3 is the dominant ST followed by ST1 and ST7. • Distribution of Blastocystis subtypes among geographic regions of Thailand differs. [ABSTRACT FROM AUTHOR]

Blastocystis is the most common protist in the gut of humans and other animals having global distribution. Occasionally, this organism has also been reported in the environment. Transmission to humans occurs via the fecal-oral route, while water also comprises a transmission route. Blastocystis has been commonly found in rivers, lakes, and wells. Nonetheless, there is limited data about the prevalence and genetic diversity of Blastocystis in tap water. The main aim of this study was to examine the presence of Blastocystis subtypes in tap water (n =20) in a community in northern Thailand. Molecular characterization using the small subunit ribosomal RNA was used to screen for Blastocystis and identify the diversity of subtypes in samples. The overall prevalence was 30% with only subtype three (ST3) encountered in the tap water. These results indicate that tap water has a potential role in the transmission of this subtype in the studied community. Further investigations should focus on expanding sampling to include additional housing complexes and screening for Blastocystis in humans who are exposed to this water. • Blastocystis occurred in 30% of tap water samples. • Blastocystis subtype 3 is the only subtype identified. • Tap water has a potential role in the transmission of Blastocystis in this community. [ABSTRACT FROM AUTHOR]

Blastocystis is a protist of controversial pathogenicity inhabiting the gut of humans and other animals. Despite a century of intense study, understanding of the epidemiology of Blastocystis remains fragmentary. Here, we aimed to explore its prevalence, stability of colonisation and association with various factors in a rural elementary school in northern Thailand. One hundred and forty faecal samples were collected from 104 children at two time points (tp) 105 days apart. For tp2, samples were also obtained from 15 animals residing on campus and seven water locations. Prevalence in children was 67% at tp1 and 89% at tp2, 63% in chickens, 86% in pigs, and 57% in water. Ten STs were identified, two of which were shared between humans and animals, one between animals and water, and three between humans and water. Eighteen children (out of 36) carried the same ST over both time points, indicating stable colonisation. Presence of Blastocystis (or ST) was not associated with body mass index, ethnicity, birth delivery mode, or milk source as an infant. This study advances understanding of Blastocystis prevalence in an understudied age group, the role of the environment in transmission, and the ability of specific STs to stably colonise children.

Blastocystis sp. is a common zoonotic intestinal protozoa which has been classified into 17 subtypes (STs). A cross-sectional study was conducted to determine the prevalence and subtype distribution of Blastocystis in villagers living on the Thai-Myanmar border, where the risk of parasitic infection is high. A total of 207 stool samples were collected and DNA was extracted. PCR and sequencing using primers targeting small-subunit ribosomal RNA (SSU rRNA) gene were performed. The prevalence of Blastocystis infection was 37.2% (77/207). ST3 (19.8% ; 41 /207) was the predominant subtype, followed by ST1 (11.6%; 24/207), ST2 (5.3%; 11/207), and ST4 (0.5%; 1/207). A phylogenetic tree was reconstructed using the maximum likelihood (ML) method based on the Hasegawa-Kishino-Yano + G + I model. The percentage of bootstrapped trees in which the associated taxa clustered together was relatively high. Some sequences of Blastocystis positive samples (TK18, 39, 46, 71, and 90) were closely related to animals (pig and cattle) indicating zoonotic risks. Therefore, proper health education in parasitic prevention for the villagers should be promoted to improve their personal hygiene. Further longitudinal studies are required to monitor the prevalence of parasitic infections after providing health education and to investigate Blastocystis ST in animals living in these villages. [ABSTRACT FROM AUTHOR]

Blastocystis is one of the most common enteric protozoa that inhabits the intestinal tract of humans and different animals. Moreover, it has a worldwide geographic distribution. Its main mode of transmission is via the fecal-oral route. At present, 26 subtypes are widely distributed across both humans and animals. The current study aimed to determine the prevalence and subtype distribution of Blastocystis among school-aged children living on the Thai-Myanmar border, Ratchaburi province, Thailand. In total, 508 samples were collected from children at six schools. The prevalence of Blastocystis infection was amplified and sequenced in the 600 bp barcode region of the small-subunit ribosomal RNA (SSU rRNA). The overall prevalence of Blastocystis infection was 3.35% (17/508). ST3 (11/17) was the most predominant subtype, followed by ST1 (5/17) and ST2 (1/17). A phylogenetic tree was constructed based on the Tamura92+G+I model using the maximum-likelihood algorithm. Surprisingly, all sequences of the ST3-positive samples were closely correlated with the cattle-derived sequence. Meanwhile, all sequences of the Blastocystis ST1-positive samples were closely correlated with the human-derived sequence. Nevertheless, further studies should be conducted to validate the zoonotic transmission of Blastocystis . Based on our findings, personal hygiene and sanitation should be improved to promote better health in children in this area.

The present study is aimed to identify the prevalence of Blastocystis subtypes isolated from patients in a major hospital in northeastern Thailand. A total of 562 stool samples were examined by culture technique, and 56 Blastocystis-positive samples were analyzed further by the combination of restriction fragment length polymorphism (RFLP) followed by polymerase chain reaction with sequence-tagged site primers (PCR-STS). By RFLP profiles, Blastocystis genotypes were categorized into four groups: group A (12, 21.4 %), group B (32, 57.1 %), group C (10, 17.9 %), and group D (2, 3.6 %). By PCR-STS, only four subtypes were identified. All 12 (21.4 %) isolates in group A were identified as subtype 1. Similarly, all 32 (57.1 %) isolates in group B were subtype 3. In group C, 10 (17.9 %) samples were all subtype 7, and two samples (3.6 %) in group D were both subtype 6. Of 56 Blastocystis-positive patients, 31 (55.4 %) were asymptomatic and 22 (39.4 %) have gastrointestinal symptoms. No significant association was observed between the Blastocystis subtypes and the clinical features. Among the Blastocystis-positive patients, the most characteristic stool samples were loose (78.6 %) and soft (17.9 %). In conclusion, the most common Blastocystis spp. in northeastern Thailand was subtype 3 followed by subtype 1. Relatively minor subtypes, subtype 6 and subtype 7 which are considered as avian subtypes, were found for the first time in humans in Thailand. [ABSTRACT FROM AUTHOR]

The article focuses on a retrospective cohort study related to the incidence and risk factors of Blastocystis infection in an orphanage in Bangkok, Thailand. As stated, the incidence rate was 1.8/100 person-months and age, nutritional status and orphans living in the room where their childcare workers were infected, were the independent risk factors associated with Blastocystis infection. Also, the conclusions indicate universal precautions like regular hand washing and careful handling of fecally contaminated materials.

Blastocystis spp. is a cosmopolitan intestinal protozoa, infecting 30 - 60% population in the developing countries worldwide. Although often asymptomatic, chronic Blastocystis spp. infection is associated with irritable bowel syndrome. The parasite displays 17 genetically diverse subtypes (STs), exhibiting a wide host-range including human, ungulates, avian, and non-human primates. In areas where human and animal live closely together, this potentially results in zoonotic infection by Blastocystis spp. This study aims to identify Blastocystis spp. subtype distribution in human and domestic animals in an endemic area for intestinal parasitic infections at the Thai-Myanmar border. Human and animal stool samples collected from six subdistricts of Tak province, Thailand, were microscopically screened for the presence of Blastocystis spp. The Blastocystis -positive samples were subjected to 18s small-subunit rRNA-based PCR amplification of the Blastocystis spp. and chain-termination sequencing. Neighbour-joining analysis was performed on the sequences for evolutional study and phylogenetic tree construction using previously published strains as reference. Forty-four samples were positive by microscopy for Blastocystis spp. and 27 samples were successfully cultured. Eight isolates from human and three from domestic animals (goats and buffalo) were PCR amplified. Multiple alignment of the DNA sequences with the subtype reference sequences and phylogenetic analysis of the 18s small-subunit rRNAs revealed ST3, a human specific subtype, as the most prevalent (6/11). Other subtypes were ST1 (1/11), ST2 (1/11), and ST13 (1/11). Two samples yielded dubious results, although one sequence from buffalo sample showed high similarity to ST15, previously reported in camels and gibbons. Two sequences of the human samples identified as ST1 (specific in human and ungulates) and ST2 (specific to human and avian) showed close resemblance to those of pigs and ducks, indicating possible zoonotic transmission risks. We provided information on the Blastocystis spp. subtypes circulating in an endemic area at the Thai-Myanmar border. The high sequence identity of Blastocystis isolates between human and domestic animal samples warrants the possible zoonotic transmission of this parasite. Our results suggest the proper policy implementation regarding water-supply and animal waste-management to prevent the parasite transmission to human. [ABSTRACT FROM AUTHOR]

Currently, the detection of human infection with Blastocystis hominis is usually based on the examination under a light microscope of faecal samples, either directly, as 'simple smears', or after some form of concentration. Whether short-term, in-vitro cultivation would increase the sensitivity of such detection remains a matter of controversy. Over 900 fresh stool specimens, from soldiers in the Royal Thai Army, were each checked for the parasite using three methods: simple smears; formalin-ethyl-acetate concentration; and cultivation in Jones' medium. Although 334 of the samples were found to be culture-positive, the parasites were only detected in 142 of the simple smears, and faecal concentration led to an even lower sensitivity (64 positive samples). In-vitro cultivation does seem worthwhile in the detection of B. hominis carriage in field studies. [ABSTRACT FROM AUTHOR]

Blastocystis is an obligate anaerobic microbial eukaryote that frequently inhabits the gastrointestinal tract. Despite this prevalence, very little is known about the extent of its genetic diversity, pathogenicity, and interaction with the rest of the microbiome and its host. Although the organism is morphologically static, it has no less than 28 genetically distinct subtypes (STs). Reports on the pathogenicity of Blastocystis are conflicting. The association between Blastocystis and intestinal bacterial communities is being increasingly explored. Nonetheless, similar investigations extending to the metabolome are non-existent.Using established NMR metabolomics protocols in 149 faecal samples from individuals from South Korea (n = 38), Thailand (n = 44) and Turkey (n = 69), we have provided a snapshot of the core metabolic compounds present in human stools with (B+) and without (B−) Blastocystis. Samples included hosts with gastrointestinal symptoms and asymptomatics. A total of nine, 62 and 98 significant metabolites were associated with Blastocystis carriage in the South Korean, Thai and Turkish sample sets respectively, with a number of metabolites increased in colonised groups. The metabolic profiles of B+ and B− samples from all countries were distinct and grouped separately in the partial least squares-discriminant analysis (PLS-DA). Typical inflammation-related metabolites negatively associated with Blastocystis positive samples. This data will assist in directing future studies underlying the involvement of Blastocystis in physiological processes of both the gut microbiome and the host. Future studies using metabolome and microbiome data along with host physiology and immune responses information will contribute significantly towards elucidating the role of Blastocystis in health and disease. [ABSTRACT FROM AUTHOR]

Blastocystis sp., the most common intestinal protozoa, remains a public health problem among people in many countries, particularly in rural areas of developing countries. The infection usually reflects poor sanitation in communities by waterborne, zoonotic, and person-to-person transmission. Interestingly, at least 17 subtypes (STs) have been reported and are associated with a broad range of animal hosts, including humans. In this study, we reported potential evidence of zoonotic transmission of Blastocystis ST1 in rural communities of eastern Thailand where the overall prevalence of Blastocystis infection was 15.7%. Two major and three minor subtypes were found to be distributed unequally in this region. Of 5 STs, only ST1 was found to be associated with pig feces in an open farm system that produced organic fertilizer for agriculture uses in the community. This finding suggests that properly protective contact and standard production of organic fertilizer from pig feces by-products could be key factors for reducing the prevalence of Blastocystis infection and prevent Blastocystis reinfection among people in the community. IMPORTANCE Blastocystis subtype 1 (ST1) in rural communities of eastern Thailand. Two major and three minor subtypes were found to be unequally distributed in this region. Interestingly, only ST1 was found to be associated with pig feces in an open farm system that produced organic fertilizer for agriculture uses in the community. The finding makes significant contributions to genetic and molecular investigations of microbial topics of practical value and suggest that properly protective contact and standard production of organic fertilizer from pig feces by-products could be key factors for reducing the prevalence of Blastocystis subtype 1 (ST1) in rural communities of eastern Thailand. Two major and three minor subtypes were found to be unequally distributed in this region. Interestingly, only ST1 was found to be associated with pig feces in an open farm system that produced organic fertilizer for agriculture uses in the community. The finding makes significant contributions to genetic and molecular investigations of microbial topics of practical value and suggest that properly protective contact and standard production of organic fertilizer from pig feces by-products could be key factors for reducing the prevalence of Blastocystis infection and prevent Blastocystis reinfection among people in the community.

Diabetes mellitus (DM) is a major global public health problem with an increasing prevalence. DM increases the risk of infections caused by bacteria, fungi, viruses, and parasites. We examined the prevalence, subtypes, and risk factors of Blastocystis infection in patients with and without DM in central Thailand. Stool samples and questionnaires were obtained from 130 people in the DM group and 100 people in the non-DM group. Blastocystis infection was identified via a nested polymerase chain reaction and subtyped via sequencing of the partial small-subunit ribosomal RNA (SSU rRNA) gene. Analysis of potential risk factors was conducted via binary logistic regression. The overall prevalence of Blastocystis infection was 10.8%, including rates of 9% and 12.3% in the non-DM and DM groups, respectively. The most prevalent subtype was ST3, followed by ST1, and ST4. Factors that potentially increased the risk of Blastocystis infection include patients being >65 years old, the presence of DM, a DM duration of ≥10 years, a low level of education, and animal ownership. In conclusion, this is the first study of Blastocystis infection in DM, and a high prevalence was found among this population. Therefore, health education promoting sanitation and hygiene is necessary to reduce and prevent infection in the community.

Blastocystis spp. is one of the most common protozoa of humans and animals worldwide. The genetic diversity of Blastocystis spp. might be associated with a wide range of symptoms. However, the prevalence of each subtype is different in each country. Until now, there is no standard method for subtyping of Blastocystis spp. We developed a sequential restriction fragment length polymorphism (RFLP) analysis for the rapid differentiation of human Blastocystis subtypes. A large-scale study was also conducted to determine the subtype distribution of Blastocystis spp. in Thailand. Stool samples were collected from 1025 school-age students in four regions of Thailand. Blastocystis infections were identified by direct smear, formalin ethyl-acetate concentration technique (FECT), Boeck and Drbohlav's Locke-Egg-Serum (LES) medium culture, and polymerase chain reaction (PCR) of small-subunit ribosomal DNA (SSU rDNA). Subtypes of Blastocystis spp. were determined by RFLP. Phylogenetic tree of partial SSU rDNA sequences of Blastocystis spp. was constructed using the Maximum Likelihood (ML) method. Out of 1025 students, 416 (40.6%) were positive for Blastocystis spp. Using two steps of RFLP reactions, we could determine subtype one–three among these students. Subtype 3 was the most common subtype (58.72%) in Thai students, followed by subtype 1 (31.2%), and subtype 2 (10.1%). Blastocystis subtype 3 was the most prevalent in all regions of Thailand. The subtype distribution of Blastocystis spp. in Thailand was different from other countries. [ABSTRACT FROM AUTHOR]

Blastocystis sp. is a common zoonotic intestinal protozoa which has been classified into 17 subtypes (STs). A cross-sectional study was conducted to determine the prevalence and subtype distribution of Blastocystis in villagers living on the Thai-Myanmar border, where the risk of parasitic infection is high. A total of 207 stool samples were collected and DNA was extracted. PCR and sequencing using primers targeting small-subunit ribosomal RNA (SSU rRNA) gene were performed. The prevalence of Blastocystis infection was 37.2% (77/207). ST3 (19.8%; 41/207) was the predominant subtype, followed by ST1 (11.6%; 24/207), ST2 (5.3%; 11/207), and ST4 (0.5%; 1/207). A phylogenetic tree was reconstructed using the maximum likelihood (ML) method based on the Hasegawa-Kishino-Yano + G + I model. The percentage of bootstrapped trees in which the associated taxa clustered together was relatively high. Some sequences of Blastocystis positive samples (TK18, 39, 46, 71, and 90) were closely related to animals (pig and cattle) indicating zoonotic risks. Therefore, proper health education in parasitic prevention for the villagers should be promoted to improve their personal hygiene. Further longitudinal studies are required to monitor the prevalence of parasitic infections after providing health education and to investigate Blastocystis ST in animals living in these villages.