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Objectives: Understanding disease transmission is a fundamental challenge in ecology. We used transmission potential networks to investigate whether a gastrointestinal protozoan (Blastocystis spp.) is spread through social, environmental, and/or zoonotic pathways in rural northeast Madagascar., Materials and Methods: We obtained survey data, household GPS coordinates, and fecal samples from 804 participants. Surveys inquired about social contacts, agricultural activity, and sociodemographic characteristics. Fecal samples were screened for Blastocystis using DNA metabarcoding. We also tested 133 domesticated animals for Blastocystis. We used network autocorrelation models and permutation tests (network k-test) to determine whether networks reflecting different transmission pathways predicted infection., Results: We identified six distinct Blastocystis subtypes among study participants and their domesticated animals. Among the 804 human participants, 74% (n = 598) were positive for at least one Blastocystis subtype. Close proximity to infected households was the most informative predictor of infection with any subtype (model averaged OR [95% CI]: 1.56 [1.33-1.82]), and spending free time with infected participants was not an informative predictor of infection (model averaged OR [95% CI]: 0.95 [0.82-1.10]). No human participant was infected with the same subtype as the domesticated animals they owned., Discussion: Our findings suggest that Blastocystis is most likely spread through environmental pathways within villages, rather than through social or animal contact. The most likely mechanisms involve fecal contamination of the environment by infected individuals or shared food and water sources. These findings shed new light on human-pathogen ecology and mechanisms for reducing disease transmission in rural, low-income settings., (© 2024 Wiley Periodicals LLC.)

Blastocystis is a common intestinal protist in humans and animals. Currently, Blastocystis infection in bar-headed geese is unknown. To understand the prevalence and distribution of Blastocystis subtypes in bar-headed geese, fecal samples were collected from 358 bar-headed geese in three regions. The total infection rate of Blastocystis in bar-headed in China was 5.9% (21/358), with 5.0% (6/120) in Aba (Ngawa) Tibetan and Qiang Autonomous Prefect, Sichuan province, 11.8% (14/119) in Maqu county, Gansu province, and 0.8% (1/119) in Caohai, Wei ning county, Guizhou province. The differences in prevalence rates by region were statistically significant. Sequences analyses showed that one known subtype (ST7, n = 16) and one potentially novel ST (n = 5) in bar-headed geese were detected in this study. This is the first report on the prevalence and subtype distribution of Blastocystis in bar-headed geese, which will improve our understanding of the epidemiology and public health implications of Blastocystis infection in wild migratory birds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Blastocystis is one of the most common zoonotic intestinal protozoa with global distribution and can cause gastrointestinal syndrome mainly characterized by diarrhea. School children are the main susceptible population. No epidemiological data on Blastocystis among school children in Hainan, the only tropical island province in China. Between March 2021 and June 2023, 1973 fecal samples were collected from school children across three regions in Hainan province. Blastocystis was examined by amplifying the small subunit ribosomal RNA (SSU rRNA) gene via polymerase chain reaction (PCR), and subtypes were identified through DNA sequencing and phylogenetic analysis. The overall prevalence of Blastocystis was 7.3 % (144/1973). The differences in infection rates across different regions, nationalities, and educational stages are statistically significant (P < 0.001). Five subtypes were identified, of which ST3 was the dominant subtype (60.4 %; 87/144), followed by ST1 (27.8 %; 40/144), ST7 (10.4 %; 15/144), ST6 (0.7 %; 1/144), and ST2 (0.7 %; 1/144). 42 known sequences and 15 novel sequences were identified including eight new variations of the ST1 (ST1-16∼ST1-23) with similarities ranging from 98.3 % to 99.78 % and seven new variations of the ST7 (ST7-7∼ST7-13) with similarities ranging from 97.7 % to 99.79 % by intra-subtype genetic polymorphisms analysis. The results evaluate the public health risks of Blastocystis among school children in Hainan and the sources of infection were discussed, providing important basic data for the effective prevention and control of intestinal parasitic diseases in Hainan., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Migratory birds play an important role in the cross-regional transmission of zoonotic pathogens. Assessing the presence of zoonotic pathogens carried by migratory birds is critical for disease control. However, information about Blastocystis infection in the migratory birds is very limited. Thus, we conducted this study with the aim to explore the occurrence, prevalence and subtyping of Blastocystis in four breeds of migratory birds in northeastern China. From October 2022 to April 2023, a total of 427 fresh fecal samples were obtained from four breeds of migratory birds in five nature reserves in northeastern China, and screened for Blastocystis by PCR amplification. Twenty-one (4.92 %) of the studied samples were confirmed Blastocystis-positive, and two known zoonotic subtypes ST6 and ST7 were founded, with ST7 being the major subtype. Until now, we firstly reported the infection status and subtyping of Blastocystis in the migratory Greater White-Fronted Goose, White Stork, Oriental White Stork and Bean Goose in China. More importantly, these findings present further data on the genetic diversity and transmission routes of Blastocystis and further arouse public health concerns about this organism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: Blastocystis, a widely distributed zoonotic protozoan infecting both humans and numerous animals, remains poorly understood with its potential medical and veterinary significance. This study examined the molecular occurrence and genetic variation of Blastocystis in children and calves in Bangladesh to explore cross-species transmission and disease burden., Methods: In total, 998 DNA samples were investigated, comprising 299 stool DNA from children and 699 fecal DNA from calves, using polymerase chain reaction and sequencing of the small subunit ribosomal RNA (SSU rRNA) gene., Results: This study detected Blastocystis in 5.35% of the children and 14.74% of the calves. While slight variations in occurrence rates were observed across different study variables, none were statistically significant. The occurrence was highest among children under 5 years and calves aged 1-3 months. Regarding breed, the Holstein Friesian cross and the Jersey cross exhibited higher rates of infection. Conversely, occurrences were lower among children and calves in Gazipur district. The remaining parameters displayed nearly equivalent percentages of Blastocystis. The subtypes identified in children included ST1, ST3, and ST4, with ST1 comprising 50% of them. ST3 and ST4 were also found in calves, alongside ST10 (55.34%) being the most prevalent. Other subtypes found in calves were ST14, ST21, and ST24-ST26., Conclusions: This study found that Blastocystis is more common in calves than in children in Bangladesh, with genetic diversity of nine subtypes. The common occurrence of identical variants of two subtypes in both populations suggests potential zoonotic transmission, highlighting the necessity for further molecular investigations and comprehensive measures within the One Health framework to mitigate public health risks., (© 2024. The Author(s).)

Blastocystis sp. and Dientamoeba fragilis are intestinal protists, which are common worldwide, but the pathogenic role of these organisms in gastrointestinal diseases is still controversial. This study aimed to investigate the frequency of Blastocystis sp. and D. fragilis in stool samples from adult patients with celiac disease (CD) by using conventional and molecular methods. A total of 75 patients with CD and 75 healthy individuals were included in this study. Fresh stool specimens collected from each individual were analyzed by conventional and molecular methods. The overall prevalence of Blastocystis sp. and D. fragilis was 41.3% (31/75) and 24% (18/75) in patients with CD, and 46.7% (35/75) and 13.3% (10/75) in healthy controls, respectively. There was no statistically significant difference in the prevalence of Blastocystis sp. and D. fragilis between CD patients and healthy individuals. Blastocystis sp. subtypes were identified in 20 CD and 16 control patients and the overall subtype distribution was observed as ST1 13.9%, ST2 30.6%, and ST3 55.6%. The prevalence of Blastocystis sp. and D. fragilis in adults with CD is similar to the prevalence of protozoa in healthy adults. In this study, the most prevalent Blastocystis subtype was ST3 and the most frequent allele was a34 in both CD patients and healthy individuals. No significant difference was found between the two groups in terms of the detection rates of Blastocystis sp. and D. fragilis, and it is thought that both protists may be colonisers of the intestinal microbiome., Competing Interests: Declaration of competing interest Not applicable., (Copyright © 2024. Published by Elsevier B.V.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: Recent studies determined that the amoeboid form of Blastocystis acts as a factor in stimulating the host's immune responses and ultimately results in urticaria and other skin disorders. The present study was conducted in order to determine the prevalence of Blastocystis in people referred to Bushehr city health centers and the relationship of this parasite with urticaria., Methods: Fecal samples were collected from 180 males and females referred to Bushehr health centers and a questionnaire containing demographic information was completed for each person. Samples were examined by preparing direct smear (wet mount) and then formalin-detergent sedimentation techniques. Data were analyzed using SPSS 22.0 software and chi-square test., Results: The results showed that 11.1% of cases infected with Blastocystis and 55% of patients with Blastocystis had various gastrointestinal symptoms. Statistical analysis showed that there was no significant relationship between infection with some demographic factors such as sex, age, literacy level and residence, but this was significant with some clinical symptoms such as itching and urticaria., Conclusion: Despite the existence of conflicting information and many ambiguities about the Blastocystis , this emerging pathogen is very important in terms of causing allergic and skin disorders in sufferers, therefore, it is necessary that patients with urticaria be evaluated for Blastocystis along with other diagnostic procedures and physicians should request a test before any medical intervention. Thus, diagnosis and treatment of these people can play an important role in improving the health of society.

Domestic dogs and cats can serve as a source of environmental contamination with Toxocara spp. and Blastocystis spp., and this represents a neglected public and veterinary health problem. We assessed the microscopic and molecular prevalence of these species in a locality in Algeria and identified the associated risk factors. The faeces of 225 dogs and 78 cats were collected in Mitidja between March and July 2022. The samples were analysed by coproscopy and by polymerase chain reaction (PCR) targeting the Internal Transcribed Spacer 2 (ITS2) and Small Subunit Ribosomal (SSU-RNA) of T. canis and Blastocystis spp. respectively. The overall microscopic prevalence of Toxocara spp. in dogs and cats was 9.78 ± 1.98% and 12.82 ± 7.42%, respectively. The rate of Blastocystis spp. was 15.11 ± 2.39% and 15.38 ± 4.08% in dogs and cats, respectively while the molecular prevalence of T. canis in dogs was 4.89 ± 1.44% and in cats 1.28 ± 1.27%; the prevalence of Blastocystis spp. was 41.78 ± 3.29% and 34.62 ± 5.39% in dogs and cats, respectively. Phylogenetic and phylogeographic analyses identified the presence of the H1 subtype of T. canis in dogs, and the ST1 subtype of Blastocystis in dogs and cats. Dogs with clinical signs were more likely to be infected with T. canis (OR 6.039, P < 0.05) than healthy dogs. This study demonstrates that dogs and cats are carriers of Toxocara spp. and Blastocystis spp. and are therefore a source of environmental contamination. Veterinarians and human health professionals should work together to implement control strategies as part of a "One Health" approach to improving animal health and reducing the risk of transmission to humans., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction: The multifactorial etiology of gastroenteritis emphasizes the need for different laboratory methods to identify or exclude infectious agents and evaluate the severity of diarrheal disease., Objective: To diagnose the infectious etiology in diarrheic children and to evaluate some fecal markers associated with intestinal integrity., Materials and Methods: The study group comprised 45 children with diarrheal disease, tested for enteropathogens and malabsorption markers, and 76 children whose feces were used for fat evaluation by the traditional and acid steatocrit tests., Results: We observed acute diarrhea in 80% of the children and persistent diarrhea in 20%. Of the diarrheic individuals analyzed, 40% were positive for enteropathogens, with rotavirus (13.3%) and Giardia duodenalis (11.1%) the most frequently diagnosed. Among the infected patients, occult blood was more evident in those carrying pathogenic bacteria (40%) and enteroviruses (40%), while steatorrhea was observed in infections by the protozoa G. duodenalis (35.7%). Children with diarrhea excreted significantly more lipids in feces than non-diarrheic children, as determined by the traditional (p<0.0003) and acid steatocrit (p<0.0001) methods. Moreover, the acid steatocrit method detected 16.7% more fecal fat than the traditional method., Conclusions: Childhood diarrhea can lead to increasingly severe nutrient deficiencies. Steatorrhea is the hallmark of malabsorption, and a stool test, such as the acid steatocrit, can be routinely used as a laboratory tool for the semi-quantitative evaluation of fat malabsorption in diarrheic children.

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., 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.

Background: Blastocystis is a human gut symbiont of yet undefined clinical significance. In a set of faecal samples collected from asymptomatic children of six distant populations, we first assessed the community profiles of protist 18S rDNA and then characterized Blastocystis subtypes and tested Blastocystis association with the faecal bacteriome community., Methods: Stool samples were collected from 244 children and young persons (mean age 11.3 years, interquartile range 8.1-13.7) of six countries (Azerbaijan 51 subjects, Czechia 52, Jordan 40, Nigeria 27, Sudan 59 and Tanzania 15). The subjects showed no symptoms of infection. Amplicon profiling of the 18S rDNA was used for verification that Blastocystis was the most frequent protist, whereas specific real-time PCR showed its prevalence and quantity, and massive parallel amplicon sequencing defined the Blastocystis subtypes. The relation between Blastocystis and the stool bacteriome community was characterized using 16S rDNA profiling., Results: Blastocystis was detected by specific PCR in 36% (88/244) stool samples and was the most often observed faecal protist. Children from Czechia and Jordan had significantly lower prevalence than children from the remaining countries. The most frequent subtype was ST3 (49%, 40/81 sequenced samples), followed by ST1 (36%) and ST2 (25%). Co-infection with two different subtypes was noted in 12% samples. The faecal bacteriome had higher richness in Blastocystis-positive samples, and Blastocystis was associated with significantly different community composition regardless of the country (p < 0.001 in constrained redundancy analysis). Several taxa differed with Blastocystis positivity or quantity: two genera of Ruminococcaceae were more abundant, while Bifidobacterium, Veillonella, Lactobacillus and several other genera were undrerrepresented., Conclusions: Asymptomatic children frequently carry Blastocystis, and co-infection with multiple distinct subtypes is not exceptional. Prevalence and quantity of the organism clearly differ among populations. Blastocystis is linked to both faecal bacteriome diversity and its composition., (© 2021. The Author(s).)

Introduction: A lack of prospective and longitudinal data on pre- and post-travel carriage of Blastocystis spp. complicates interpretation of a positive test post-travel. Therefore we studied dynamics of Blastocystis carriage in a cohort of Dutch travellers., Methods: From the prospective, multicentre COMBAT study among 2001 Dutch travellers, a subset of 491 travellers was selected based on travel destination to 7 subregions (70 or 71 travellers each). Faecal samples taken directly before and after travel were screened for Blastocystis with qPCR, followed, when positive, by sequence analysis to determine subtypes., Results: After exclusion of 12 samples with missing samples or inhibited qPCR-reactions, stool samples of 479 travellers were analysed. Before travel, 174 of them (36.3%) carried Blastocystis and in most of these, the same subtype was persistently carried. However, in 48/174 of those travellers (27.6%; CI95 20.8-36.6%) no Blastocystis or a different subtype was detected in the post-travel sample, indicating loss of Blastocystis during travel. Only 26 (5.4%; CI95 3.7%-8.0%) of all travellers acquired Blastocystis, including two individuals that were already positive for Blastocystis before travel but acquired a different subtype during travel., Discussion: This study shows that Blastocystis carriage in travellers is highly dynamic. The observed acquisition and loss of Blastocystis could either be travel-related or reflect the natural course of Blastocystis carriage. We demonstrate that the majority of Blastocystis detected in post-travel samples were already carried before travel., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Polymerase chain reaction (PCR) is an effective technique for diagnosis of Blastocystis infection. Notably, DNA isolation procedure is extremely critical for the PCR step. In the present study, a recently described extraction procedure, named as the "sand method" was modified and adapted for isolation of Blastocystis DNA. To evaluate its efficacy, the current method and QIAamp DNA Stool Mini Kit (Qiagen) were applied to fresh human stool samples. Our results indicated that, the mean DNA concentrations obtained by the sand method and the commercial kit were 48 and 55 ng/μl, respectively. Also, no DNA inhibitors were detected in two methods. The sand method was capable of detecting 16 parasites per 50 mg feces. DNA samples extracted by both methods were subjected to PCR. Blastocystis spp. were detected in 11 (31.4%) of 35 samples, and perfect agreement (κ: 1.000) was found between the PCR-sand method and PCR-commercial kit method. The samples that were detected positive by PCR-sand method were successfully sequenced, and Blastocystis subtypes (STs) were identified as ST3, ST2 and ST1. In conclusion, the present study indicates that the sand method provides a simple, rapid and inexpensive procedure for reliable extraction of Blastocystis DNA from stool samples., (Copyright © 2018 Elsevier Inc. All rights reserved.)

There are inconsistent findings about the role of Blastocystis infection in irritable bowel syndrome (IBS). The present study was aimed to determine the frequency of Blastocystis and their subtypes (ST) in patients with IBS. A total of 122 patients with IBS and 122 healthy individuals referred to the medical laboratory centers in Ahvaz (southwest of Iran) participated in the study. The frequency of Blastocystis was determined. Blastocystis genomic DNA was extracted from positive feces, and PCR was performed using seven primer pairs targeting the SSU rDNA gene. Blastocystis was detected in 19.67% of patients with IBS and 17.2% of individuals without IBS. The difference between two groups was not statistically significant (P=0.3). Among the five subtypes of Blastocystis, ST3 was more common in patients with IBS and control group. However, there were no significant differences between two groups in terms of subtypes of Blastocystis (P=0.6). It seems, the role of Blastocystis in the etiology of IBS should be further investigated. Furthermore, a model of study should be designed to investigate the role of host factors in severity of parasitic disease., (Copyright © 2017. Published by Elsevier B.V.)

Blastocystis spp. is the most common enteric protist found in human feces. The pathogenic role of Blastocystis remains controversial and it has been suggested that the symptomatology of Blastocystis is associated with its subtypes (ST). However, only few studies have investigated the relationship between the symptomatology and subtypes of Blastocystis in children. This study aimed to investigate the prevalence of Blastocystis in children aged 3 to 13years with or without gastrointestinal complaints and determine the distribution of the subtypes of Blastocystis. A total of 303 stool samples obtained from symptomatic (n=84) and asymptomatic (n=219) children were included in the study. The presence of Blastocystis was investigated using native-lugol examination, trichrome staining and real-time PCR method. Using the real-time PCR method, 115 samples were found positive for Blastocystis. Subtyping was successfully performed on 46 samples using sequenced-tagged site (STS) primers and PCR. The remaining 69 samples could not be subtyped. The most frequently detected subtype was ST3 (43.4%) followed by ST1 (26.1%), ST4 (10.9%) and ST2 (8.7%). The mixed subtypes were identified in five samples (10.9%) as; ST1+ST3 (n=3), ST1+ST2 (n=1) and ST2+ST3 (n=1). None of the samples had ST5, ST6 or ST7. No statistically significant difference was found between the symptomatic and asymptomatic groups in terms of the Blastocystis positivity and the distribution of subtypes (p>0.05). To our knowledge, this is the first study to investigate the subtype distribution of Blastocystis in children in Turkey and the results are in agreement with the related data available in Turkey., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Background: In the last decades, the enormous influx of immigrants to industrialized countries has led to outbreaks of parasitic diseases, with enteric infections being amongst the most frequently encountered. In its strategy to control such infection, Qatar has established the Pre-Employment Certificate (PEC) program which requires medical inspection before arrival in Qatar and which is mandatory for immigrant workers travelling to the country. To assess the reliability of the PEC, we conducted a survey of intestinal parasites, based on examination of stool samples provided by immigrant workers (n = 2,486) recently arrived in Qatar., Results: Overall prevalence of helminths was 7.0% and that of protozoa was 11.7%. Prevalence of combined helminths was highest among the western Asians and the highest prevalence of combined protozoan parasites was among workers from North to Saharan Africa. Analysis of temporal changes showed an increasing trend of protozoan infections over the investigated 3 years. A major contribution to this temporal change in prevalence came from Blastocystis hominis as well as from other protozoan species: Giardia duodenalis and Endolimax nana. Analysis of the temporal trend in species richness of the protozoan species showed a significant increase in the mean number of species harboured per subject across this period., Conclusion: The increase of protozoan infections over recent years raises some concerns. It suggests that screening protocols for applicants for visas/work permits needs to be revised giving more careful attention to the intestinal protozoan infections that potential immigrants may harbor.

Blastocystis is estimated to be one of the most common parasites of the intestinal tract of humans, comprising multiple subtypes (ST). Meanwhile, the distribution of Blastocystis ST in many communities and countries remains unknown. In the present work, we aimed to identify the prevalence of Blastocystis and the ST distribution in human stool samples collected from healthy expatriates from different geographical regions and residing in Sharjah, United Arabian Emirates (UAE). A total of 133 samples were screened and subtyped using partial small subunit ribosomal RNA gene sequencing. Fifty-nine (44.4%) samples were identified as positive. Among these, 39 were successfully sequenced and subtyped. The ST distribution was as follows: ST3, 58.9% (23/39); ST1, 28.2% (11/39); and ST2, 7.6% (3/39). No correlation between geographic origin and infection (χ(2)=11.006; P=0.528) nor gender and infection (χ(2)=1.264; P=0.261) was observed. The data were compared with those available for other Middle Eastern and North African neighboring countries. This study is the first to provide data concerning the prevalence of Blastocystis and the frequency of various STs in the UAE, confirming the absence of ST4 and the commonness of ST1, ST2, and ST3 in this geographical region., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The performance of a multiplex real-time PCR for the detection of Blastocystis, Dientamoeba fragilis, Giardia lamblia, Cryptosporidium species and Entamoeba species in faecal samples was evaluated in an observational prospective study. Paediatric patients (0-18 years) presenting with gastrointestinal symptoms and suspected of having enteroparasitic disease were included. A questionnaire on gastrointestinal symptoms and the chosen treatment was completed at the start of the study and after 6 weeks. Of 163 paediatric patients (mean age, 7.8 years), 114 (70%) had a PCR-positive faecal sample. D. fragilis was detected most frequently, in 101 patients, followed by Blastocystis in 49. In faecal samples of 47 patients, more than one protozoan was detected, mainly the combination of D. fragilis and Blastocystis. Reported gastrointestinal symptoms were abdominal pain (78%), nausea (30%), and altered bowel habits (28%). Eighty-nine of the PCR-positive patients were treated with antibiotics. A significant reduction in abdominal pain was observed both in treated and in untreated patients. This study demonstrated that multiplex real-time PCR detects a high percentage of intestinal protozoa in paediatric patients with gastrointestinal symptoms. However, interpretation and determination of the clinical relevance of a positive PCR result in this population are still difficult., (© 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.)

Background: Blastocystis has been associated with various symptoms of the gastrointestinal tract. We aimed to investigate the prevalence of Blastocystis in children with celiac disease (CeD) or functional abdominal pain (FAP) and to evaluate its subtypes (STs) with respect to demographic, socioeconomic and epidemiological factors. Methods: Overall, 161 fecal samples were collected from healthy children and patients with FAP or CeD in Hitit University Erol Olçok Research and Training Hospital, Corum, Turkey between 2016-2018. Samples were examined using both native-Lugol (NL) and trichrome-stained (TS) smears, and further analyses by PCR and Sanger sequencing were performed. A standard questionnaire was applied to obtain demographic, socioeconomic, epidemiological data. Results: Blastocystis was found in 10.6% of the total study population. Neither bacteria nor any other parasites were found, except for one Giardia (0.6%) in the CeD group. The presence/absence of the parasite was not found to be associated with demographic, socioeconomic and epidemiological factors. Blastocysis was detected in 11.5% (6/52) of the CeD, 7.7% (4/52) of the FAP, and 12.3% (7/57) of the healthy group. Diagnostic methods were similar in terms of Blastocystis detection (P= 0.671), and there was fair agreement between the NL, TS and PCR (Fleiss' Kappa=0.847, P=0.001). ST2 (42.8%) and ST3 (35.7%) were the predominant STs followed by ST1 (21.4%). Conclusion: We observed no difference between study groups in terms of Blastocystis prevalence. ST1, ST2 and ST3 subtypes were detected. Blastocystis prevalence and STs were not related to any of the demographic, socioeconomic and epidemiological factors. [ABSTRACT FROM AUTHOR]

Purpose: Blastocystis is a common enteric human parasite of non-conclusive pathogenicity which may be determined by subtype (ST) variation. Colorectal cancer (CRC) is considered one of the primary causes of cancer mortality. Blastocystis ST7 has been shown to reduce beneficial intestinal microbiota and may exacerbate CRC. This study assessed the possible association between Blastocystis STs and CRC in comparison to non-cancer patients. Material and Methods: A total of 200 fecal samples were obtained from CRC (100) and non-CRC (100) individuals attending Beni-Suef University Hospital, Egypt. Blastocystis was searched for in all samples using microscopy and culturing. Positive subculture samples were genetically sequenced and subtyped using conventional polymerase chain reaction (PCR). Blastocystis STs were determined by sequencing and a phylogenetic tree was created. Related patient characteristics and tumor stages were analyzed for association with presence of Blastocystis. Results: Blastocystis was identified in 52% and 42% of CRC and non-cancer individuals, respectively. ST1, 2, and 3 were isolated from both cancer and non-cancer individuals; however, for the first time, ST7 was only isolated from CRC stool samples with significant association. Associated patient characteristics were evaluated as predictors. Conclusion: Blastocystosis is highly prevalent in CRC patients, predominantly in the latest CRC grades and stages. To the best of our knowledge, this is the first study to report the identification of Blastocystis ST7 in CRC patients. To determine whether certain STs of Blastocystis are associated with CRC would require further research, including the role played by gut microbiota. [ABSTRACT FROM AUTHOR]

The role played by Blastocystis in humans has been a subject of discussion due to its intestinal effects and modifications in the intestinal microbiota. We aimed to analyze the relationship between Blastocystis subtypes ST1-4 and 7, the Firmicutes to Bacteroidetes ratio (F/B ratio) of fecal microbiota, and chronic stress in university students. This study had a cross-sectional design with a sample of 202 students. We analyzed fecal and hair samples, and stress inventories were applied to the students. The results showed a frequency of Blastocystis-colonized students of 52.97%. Regarding fecal microbiota, a median RAU of 0.801 for Firmicutes and 0.82 of Bacteroidetes were obtained, with an F/B ratio of 0.83. A low F/B ratio (66.04%) was more frequent in Blastocystis-colonized students, whereas a high F/B ratio (68.09%) (p = < 0.0001) was found in the Blastocystis-non-colonized. Only Blastocystis ST3 did not significantly correlate with a low F/B ratio (p = 0.290). The ST4 was associated with lower values of cortisol (p = 0.030), psychological stress (p = 0.040), and lower frequency of constipation (p = 0.010). Only two students with the ST1 had abdominal pain (p = 0.007). Our results suggest that colonization by Blastocystis subtypes can modify the intestinal microbiota due to a decreased ratio between the two most representative phyla (F/B). Also, the results of this study show that ST4 colonization is related to a lower level of chronic stress. [ABSTRACT FROM AUTHOR]

Background: Intestinal parasites (IPs) are widely distributed worldwide and are one of the major contributors to gastrointestinal disease. Their prevalence is associated with poor access to water, sanitation and hygiene (WASH). The objective of this study was to identify the prevalence of IPs, including soil-transmitted helminths (STH), and their relation to socioeconomic characteristics, as well as a first approach to molecularly characterize the types of Giardia intestinalis, Blastocystis spp. and Entamoeba histolytica present in an indigenous community from Puerto Iguazú, Misiones, Argentina. Methods: A cross-sectional study was conducted in the rural settlement of Fortin Mbororé between January and March 2018. Socioeconomic variables, household characteristics, and stool and blood samples were collected. Standard coprological techniques were used to analyze stool samples, and a complete hemogram was performed on the blood samples. Giardia intestinalis microscopy-positive samples were genetically typed by the β-giardin (bg) gene. Molecular identification of Blastocystis spp. subtypes and E. histolytica were carried out by amplification and sequencing of a partial fragment of the small subunit ribosomal RNA gene (SSU rDNA). Results: The overall prevalence of IPs was 92.7%, with 72.0% specifically for hookworm. IPs were significantly more prevalent in preschool- and school-age children (P < 0.05). No formal education (P = 0.035), the presence of unimproved floors (P = 0.001) and overcrowding (P = 0.005) were significantly associated with IP infection. Hookworm was associated with anemia (P = 0.019). Molecular characterization revealed the presence of E. histolytica sub-assemblages AII (12.5%), AIII (87.5%) and BIV (100%); one case of sub-assemblage D for G. intestinalis; and the presence of subtypes ST1 (14.8%), ST2 (14.8%) and ST3 (70.4%) of Blastocystis spp. Conclusions: Protozoans detected in this study are transmitted mainly through water contaminated with fecal matter, evidencing the need to improve the quality of water and sanitation for the inhabitants of Fortín Mbororé. Molecular characterization showed that domestic animals can be implicated in the zoonotic transmission of G. intestinalis and Blastocystis spp. to humans. A hyperendemic area for STH was found, with hookworm prevalence greater than 50%. Therefore, improvements in WASH as well as mass deworming programs need to be implemented in this area to control and decrease the prevalence of IPs in general and STH in particular. [ABSTRACT FROM AUTHOR]

Blastocystis is the most frequently isolated protozoan from human stool. Its role in human health is still debated, and a high prevalence was reported in irritable bowel syndrome (IBS) subjects, suggesting a potential link with microbiota. In the present study, we aimed to investigate prokaryotic and eukaryotic microbiota in both IBS-C (constipated) and healthy individuals. We recruited 35 IBS-C patients and 23 healthy subjects, from which 12 and 11 carried Blastocystis , respectively. We performed 16S and 18S rRNA high-throughput sequencing on feces. Whereas we did not observe differences between infected and non-infected controls, several phyla were significantly modified in IBS-C patients according to the presence of Blastocystis. Tenericutes phylum and Ruminococcaceae family were especially increased in Blastocystis carriers. Furthermore, colonization with Blastocystis was associated with discrete changes in the microbial eukaryome, particularly among the Fungi taxa. Depending on the group of patients considered, the mycobiota changes do not go in the same direction and seem more deleterious in the IBS-C group. These results encourage further in vivo and in vitro investigations concerning the role of Blastocystis in the gut environment. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to evaluate the results of examination for intestinal parasites in fecal samples sent to our laboratory by obtaining from patients applied to the hospital because of various complaints. Methods: 12 239 stool samples were examined by one or several methods such as direct wet mount using 1:5 dilution of Lugol's iodine solution, sedimentation technique using fecal concentration tubes, Cryptosporidium/Giardia immunofluorescence (Crypto/Giardia Cel, Cellabs, Brookvale, NSW, Australia), ELISA for detecting Entamoeba histolytica adhesin antigen (E. histolytica II™, TechLab, Blacksburg, VA, USA), multiplex polymerase chain reaction for E. histolytica, Giardia intestinalis, Cryptosporidium spp. and Dientamoeba fragilis (Multiplexed Diagnostics Gastrointestinal Parasites, 5Plex, AusDiagnostics, Mascot, NSW, Australia) between January 2015 and July 2018. Results: In 683 (5.58%) of fecal samples, parasites were detected by any method. Number of patients with parasitic infection were higher in summer (n=208), followed by autumn (n=165), winter (n=163) and spring (n=147), respectively. Conclusions: It is thought that parasitic infections involving the gastrointestinal tract are still important for public health; direct microscopy alone is not sufficient for the diagnosis, and the diagnostic possibility can be increased by supporting with immunological and molecular methods. It should be kept in mind that bacterial and viral agents as well as parasitic infectious agents should be investigated in patients presenting with gastrointestinal symptoms especially in summer. [ABSTRACT FROM AUTHOR]

Blastocystis is a parasite with a worldwide distribution and a varying prevalence in different countries. The pleomorphic nature of the protozoon and the lack of understanding a possible pathogenesis have led to confusion regarding its clinical significance. The aim of the study was to shed light on clinical characteristics of pediatric patients in Swiss children with a positive stool sample for Blastocystis, in order to provide recommendations for a practical approach for the clinician to know whom, when, and how to test. This is a retrospective study of pediatric patients, whose stool has been tested positive for Blastocystis in the last 10 years in northern Switzerland. A total of 4047 stool samples, belonging to 1887 different patients, were analyzed; 240 stool samples (of 160 patients) were tested positive for Blastocystis. On average, 2.2 (CI 1.98-2.35) stool samples per patient were analyzed, of which 1.48 (CI 1.36-1.61) were positive for Blastocystis. In 63% abdominal pain was the leading symptom, while in 17.5% it was an accidental finding without symptoms. There was a high significance in correlation of abdominal pain and chronicity (p < 0.0001) but none in diarrhea (p = 0.082) nor nausea/vomiting or other symptoms and chronicity. Followed by Entamoeba coli (8%), 26.3% of the patients with Blastocystis had a co-infection with another parasite, mostly Endolimax nana (13%).Conclusion: Carriage of Blastocystis is common; therefore, only children/teenagers at risk for a symptomatic Blastocystis infection should be tested. There is a good correlation between Blastocystis and chronic abdominal pain. Children with abdominal symptoms persisting over 4 weeks should have two different stool samples analyzed. No screening after travels/immigration is recommended.What is Known:• Blastocystis has a worldwide distribution.• The clinical significance is unclear.What is New:• Based on retrospective data, we recommend to only test children/teenagers with chronic abdominal pain for Blastocystis.• Two different stool samples should be examined by microscopy; serological investigations are not warranted. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to determine the molecular characterization and subtype distribution of Blastocystis sp. isolated from cancer children and adolescents in Shiraz, Fars province, southwestern Iran. Methods: Overall, 200 fecal samples obtained from cancer children and adolescents under 18 years old (107 males and 93 females) and checked by microscopy, culture, and molecular methods (PCR). Possible etiological factors and clinical characteristics of Blastocystis infection were also evaluated and compared between Blastocystis infected and non-infected patients. Results: Thirteen of 200 (6.5%) stool samples were positive for Blastocystis by microscopy. While 21 of 200 (10.5%) were positive by culture, and 24 of 200 (12%) were positive by PCR. Out of 24 positive samples tested by PCR and sequencing, ST3 was reported as the most common subtype (nine samples, 37.5%), followed by ST2 (eight samples, 33.3%), ST1 (five samples, 20.9%), and ST7 (two samples, 8.3%). The prevalence of Blastocystis infection in males was significantly higher than females (p = 0.024). Also, Blastocystis was more prevalent in patients who had received at least eight chemotherapy cycles than fewer (p = 0.002). However, no associations were found between Blastocystis-positive rate and age, residence, type of cancers, or contact with animals. Also, there was no significant difference between frequency of Blastocystis subtypes in symptomatic and asymptomatic cancer patients. Conclusions: Various controlled epidemiologic and topographic studies need to confirm or reject these possible associations with Blastocystis infection. The data from this study are an invaluable addition to the growing body of research studies on Blastocystis infection in cancer patients. [ABSTRACT FROM AUTHOR]

Blastocystis is one of the most commonly detected genera of protozoan parasites in the human intestines as well as the intestines of many other species such as pigs in several geographical regions worldwide. However, no studies have examined Blastocystis in pigs in Korea. In this study, PCR and nucleotide sequencing were performed to evaluate the genetic diversity and zoonotic potential of Blastocystis using pig fecal samples. We obtained 646 stool samples from groups of piglets, weaners, growers, finishers, and sows in Korea. A total of 390 Blastocystis-positive samples were identified, and the infection rate was 60.4%. The infection rates were significantly related to age and region. The 4 subtypes (STs) of Blastocystis confirmed by phylogenetic analysis were ST1, ST2, ST3, and ST5, indicating the high genetic diversity of Blastocystis in Korean pigs. ST5 was highly distributed in Korean pigs among detected STs in this study. Some sequences were closely related to those of Blastocystis isolated from humans. This is the first study of Blastocystis in pigs in Korea. Based on the results, Blastocystis is prevalent in Korean pigs. Although a small number of samples were obtained in some areas, the clinical development of Blastocystis infection in pigs and potential for human transmission should be further examined. [ABSTRACT FROM AUTHOR]

• Full-length 18S rRNA sequences were extracted from RNA-derived metagenomics datasets. • No primer bias hampers the analysis of data generated using unbiased metagenomics. • Ten different parasite species/subtypes (ST) were detected in 37 datasets. • Blastocystis ST 15 was discovered for the first known time in swine faeces. • Metagenomics has the potential to be used as a generic pathogen detection tool. Efficient and reliable identification of emerging pathogens is crucial for the design and implementation of timely and proportionate control strategies. This is difficult if the pathogen is so far unknown or only distantly related with known pathogens. Diagnostic metagenomics – an undirected, broad and sensitive method for the efficient identification of pathogens – was frequently used for virus and bacteria detection, but seldom applied to parasite identification. Here, metagenomics datasets prepared from swine faeces using an unbiased sample processing approach with RNA serving as starting material were re-analysed with respect to parasite detection. The taxonomic identification tool RIEMS, used for initial detection, provided basic hints on potential pathogens contained in the datasets. The suspected parasites/intestinal protists (Blastocystis, Entamoeba, Iodamoeba, Neobalantidium, Tetratrichomonas) were verified using subsequently applied reference mapping analyses on the base of rRNA sequences. Nearly full-length gene sequences could be extracted from the RNA-derived datasets. In the case of Blastocystis , subtyping was possible with subtype (ST)15 discovered for the first known time in swine faeces. Using RIEMS, some of the suspected candidates turned out to be false-positives caused by the poor status of sequences in publicly available databases. Altogether, 11 different species/STs of parasites/intestinal protists were detected in 34 out of 41 datasets extracted from metagenomics data. The approach operates without any primer bias that typically hampers the analysis of amplicon-based approaches, and allows the detection and taxonomic classification including subtyping of protist and metazoan endobionts (parasites, commensals or mutualists) based on an abundant biomarker, the 18S rRNA. The generic nature of the approach also allows evaluation of interdependencies that induce mutualistic or pathogenic effects that are often not clear for many intestinal protists and perhaps other parasites. Thus, metagenomics has the potential for generic pathogen identification beyond the characterisation of viruses and bacteria when starting from RNA instead of DNA. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • First epidemiological survey of Blastocystis sp. in domestic cattle in Lebanon. • Identification of the parasite with a high prevalence in livestock cattle. • Confirmation of ST10 and ST14 as cattle-adapted STs. • High infection rate of Blastocystis sp. in dairy farmers and hospitalized patients. • Negligible role played by cattle as zoonotic reservoirs for the parasite. Abstract Blastocystis sp. is frequently identified in a wide range of animal hosts, including bovids. Because of its burden and zoonotic potential, this parasite has been sought in domestic cattle from various countries, since this livestock may also represent a possible reservoir of human infection. However, epidemiological data regarding the prevalence and ST distribution of Blastocystis sp. in this animal group is lacking in Lebanon. Therefore, faecal samples were collected from a total of 254 dairy cattle raised on 55 farms located in the North Lebanon region and screened for the presence of the parasite by quantitative real-time PCR. The overall prevalence of Blastocystis sp. was shown to reach 63.4% in cattle livestock. Sequence analysis of positive samples indicated the presence of seven STs, with predominance of ST10 (44.0%) and ST14 (36.8%) and lower proportions of ST2 (8.0%), ST1 (7.2%), ST5 (2.4%), ST3 and ST7 (0.8% each). This survey was the first conducted worldwide reporting ST2 and ST7 in domestic cattle and confirmed that ST10 and ST14 represent cattle-adapted STs in view of their high prevalence. Faecal samples from in-contact dairy farmers and patients hospitalised in the same Lebanese governorate who reported no contact with cattle livestock were also analysed for the presence of Blastocystis sp. The same three STs were identified in both human cohorts, with predominance of ST3, followed either by ST1 or ST2 depending of the group. No other STs, including ST10 or ST14, have been reported. Moreover, even though ST1, ST2 and ST3 were found to be common to dairy cattle and farmers cohorts, only one ST3 isolate showed 100% sequence identity between both hosts. Consequently, the presence and low prevalence of ST1, ST2, ST3, ST5 and ST7 identified herein in domestic cattle, most of which exhibit low host specificity, could be derived from occasional direct exposure to faecal material from human and non-human hosts or by ingestion of contaminated drinking water or food in the enclosure of the farms. Together with the absence of ST10 and ST14 in the human population, these data suggest that cattle play a negligible role as zoonotic reservoirs of Blastocystis sp. [ABSTRACT FROM AUTHOR]

Objectives: To determine the prevalence and risk factors associated with intestinal parasites in the population of San Juan Cosala, Jalisco, Mexico. Methods: A total of 277 samples from 104 participants were analysed using direct smear, flotation, formaldehyde/ethyl acetate, and modified Kinyoun's acid-fast stain methods. The Graham method was applied only for samples from children under 12 years of age for the diagnosis of Enterobius vermicularis. Results: The prevalence of parasite infections in the study population was 77.9% including: Entamoeba histolytica/E. dispar/E. moshkovskii/E. bangladeshi (37.5%), Giardia intestinalis (11.5%); commensals: Endolimax nana (44.2%), Entamoeba coli (27.9%), Chilomastix mesnili (6.7%) and Iodamoeba bütschlii, (2.9%); emerging intestinal protozoans: Blastocystis spp. (49%), Cryptosporidium spp. (7.7%) and Cyclospora cayetanensis (2.9%); and helminths: Enterobius vermicularis (18.3%) and Ascaris lumbricoides (5.8%). The results also showed that 58.64% of the studied population presented polyparasitism. A significant association was found between protozoan infections and housewives, and houses that were not built with concrete ceilings, brick walls and cement floors (p < 0.05). Conclusion: Polyparasitism was observed in over half the study population. The most prevalent parasite was Blastocystis spp, whilst the prevalence of helminths was less than that of protozoans. The risk factors for infection to intestinal parasites were being a housewife and not having solid brick, cement and concrete materials for house construction. [ABSTRACT FROM AUTHOR]