BACKGROUND: Lactiplantibacillus plantarum HEAL9 and Lacticaseibacillus paracasei 8700:2 positively affect the fecal bacteriome in children with celiac disease autoimmunity after 6 months of supplementation. The aim of the present investigation was to study the effects of Lactiplantibacillus plantarum HEAL9 and Lacticaseibacillus paracasei 8700:2 on the single-cell parasitome, with a primary focus on Blastocystis. METHODS: Stool samples were collected from 78 Swedish children with celiac disease autoimmunity participating in a randomized, double-blind, placebo-controlled clinical trial to either receive a mixture of supplementation with L. plantarum HEAL9 and L. paracasei 8700:2 (n = 38) or placebo (n = 40). A total of 227 stool samples collected at baseline and after 3 and 6 months of intervention, respectively, were retrospectively analyzed for Blastocystis by quantitative real-time PCR and subtyped by massively parallel amplicon sequencing. Other single-cell parasites were detected by untargeted 18S rDNA amplicon sequencing and verified by real-time PCR. The relation between the parasites and the bacteriome community was characterized by using 16S rDNA profiling of the V3-V4 region. RESULTS: Three different single-cell protists were identified, of which the highest prevalence was found for Dientamoeba fragilis (23.1%, 18/78 children), followed by Blastocystis (15.4%, 12/78) and Entamoeba spp. (2.6%, 2/78). The quantity of the protists was stable over time and not affected by probiotic intervention (P = 0.14 for Blastocystis, P = 0.10 for D. fragilis). The positivity of the protists was associated with increased bacteriome diversity (measured by multiple indices, P < 0.03). Bacterial composition was influenced by the presence of the protists: positivity of Blastocystis was inversely associated with Akkermansia (at the levels of the genus as well as its family, order, class and phylum); P < 0.002), Faecalibacterium (P = 0.003) and Romboutsia (P = 0.029); positivity of D. fragilis was inversely associated with families Enterobacteriaceae (P = 0.016) and Coriobacteriaceae (P = 0.022) and genera Flavonifractor (P < 0.001), Faecalibacterium (P = 0.009), Lachnoclostridium (P = 0.029), Ruminococcus (P < 0.001) and Granulicatella (P = 0.018). CONCLUSIONS: The prevalence of single-cell protists is low in children with celiac disease autoimmunity. The colonization was stable regardless of the probiotic intervention and associated with increased diversity of the fecal bacteriome but inversely associated with some beneficial bacteria.
- MeSH
- autoimunita MeSH
- Bacteria MeSH
- Blastocystis * genetika MeSH
- celiakie * MeSH
- dítě MeSH
- dvojitá slepá metoda MeSH
- feces parazitologie MeSH
- Lacticaseibacillus MeSH
- Lactobacillus paracasei * MeSH
- lidé MeSH
- probiotika * terapeutické užití farmakologie MeSH
- retrospektivní studie MeSH
- ribozomální DNA MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- randomizované kontrolované studie MeSH
Blastocystis is the most prevalent microbial eukaryote in the human and animal gut, yet its role as commensal or parasite is still under debate. Blastocystis has clearly undergone evolutionary adaptation to the gut environment and possesses minimal cellular compartmentalization, reduced anaerobic mitochondria, no flagella, and no reported peroxisomes. To address this poorly understood evolutionary transition, we have taken a multi-disciplinary approach to characterize Proteromonas lacertae, the closest canonical stramenopile relative of Blastocystis. Genomic data reveal an abundance of unique genes in P. lacertae but also reductive evolution of the genomic complement in Blastocystis. Comparative genomic analysis sheds light on flagellar evolution, including 37 new candidate components implicated with mastigonemes, the stramenopile morphological hallmark. The P. lacertae membrane-trafficking system (MTS) complement is only slightly more canonical than that of Blastocystis, but notably, we identified that both organisms encode the complete enigmatic endocytic TSET complex, a first for the entire stramenopile lineage. Investigation also details the modulation of mitochondrial composition and metabolism in both P. lacertae and Blastocystis. Unexpectedly, we identify in P. lacertae the most reduced peroxisome-derived organelle reported to date, which leads us to speculate on a mechanism of constraint guiding the dynamics of peroxisome-mitochondrion reductive evolution on the path to anaerobiosis. Overall, these analyses provide a launching point to investigate organellar evolution and reveal in detail the evolutionary path that Blastocystis has taken from a canonical flagellated protist to the hyper-divergent and hyper-prevalent animal and human gut microbe.
- MeSH
- Blastocystis * genetika MeSH
- Eukaryota MeSH
- lidé MeSH
- mitochondrie genetika metabolismus MeSH
- organely metabolismus MeSH
- střevní mikroflóra * genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Výskyt střevních parazitů se v populaci, dle Informačního systému infekčních nemocí, snižuje. Záchyt je ovlivněn správností odběru stolice a metodou, kterou je vzorek vyšetřován. Na základě molekulárně biologické (PCR) diagnostiky vzorků stolic u 200 dětí jsme ukázali, že prevalence parazitóz je vyšší, než popisují dostupná data. Klinické symptomy a pozitivita výsledků v naší studii nekorelují. Eradikaci asymptomatických jedinců je třeba vždy zvážit, vzhledem k objevujícím se důkazům, že existuje nepřímá úměra mezi prevalencí střevních parazitóz a výskytem alergií, autoimunit, Crohnovy nemoci aj., stejně jako k nejasné patogenitě daných mikroorganismů.
The incidence of intestinal parasites in the population is decreasing according to the Information System of Infectious Diseases. Detection is affected by the accuracy of stool collection and the method by which the sample is examined. Based on PCR diagnostics of stool samples from 200 children, we showed that parasitosis prevalence is higher than the data described. Clinical symptoms and positivity in our study do not correlate. Eradication of asymptomatic individuals should always be considered given the emerging evidence that there is an indirect relationship between the prevalence of intestinal parasitosis and the incidence of allergies, autoimmunity, Crohn's disease, etc., as well as the unclear pathogenicity of some of the microorganisms.
- MeSH
- Blastocystis hominis izolace a purifikace MeSH
- blastocystóza diagnóza epidemiologie terapie MeSH
- dientamébóza diagnóza epidemiologie terapie MeSH
- dítě MeSH
- enterobióza diagnóza epidemiologie terapie MeSH
- Enterobius izolace a purifikace MeSH
- epidemiologické studie MeSH
- lidé MeSH
- mladiství MeSH
- parazitární nemoci střev * diagnóza epidemiologie terapie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
Species of Blastocystis Alexieff, 1911 are anaerobic intestinal protists found in humans and many kinds of animals that mainly cause diarrhea, abdominal pain and other clinical symptoms. At present, data on the prevalence and subtype diversity of species of Blastocystis in domestic rabbits are very limited. The purpose of this study was to characterise the infection rate and gene subtype distribution of Blastocystis sp. in domestic rabbits in Henan Province, Central China, and provide foundation for prevention and control of the disease caused by Blastocystis sp. in domestic rabbits. DNA was extracted from 286 fresh rabbit faecal samples collected from four areas of Henan Province, Central China. All DNA samples were screened using PCR and positive samples were sequenced to identify individual subtypes based on the small ribosomal subunit (SSU rRNA) gene. The overall infection rate of Blastocystis sp. in domestic rabbits in Henan Province was 15% (43/286). Three subtypes were identified, including ST1 (26/43, 60%), ST3 (5/43, 12%) and ST7 (12/43, 28%), all of which belonged to potentially zoonotic subtypes, ST1 was the dominant gene subtype. These results showed that infection with Blastocystis sp. was common in domestic rabbits in Henan Province, Central China, and was represented by zoonotic subtypes. Therefore, special attention should be paid to reduce the risk of transmission of Blastocystis sp. from domestic rabbits to humans.
Blastocystis is the most commonly found intestinal protist in the world. Accurate detection and differentiation of Blastocystis including its subtypes (arguably species) are essential to understand its epidemiology and role in human health. We compared (i) the sensitivity of conventional PCR (cPCR) and qPCR in a set of 288 DNA samples obtained from stool samples of gut-healthy individuals, and (ii) subtype diversity as detected by next-generation sequencing (NGS) versus Sanger sequencing. Real-time PCR resulted in more positive samples than cPCR, revealing high fecal load of Blastocystis based on the quantification curve in most samples. In subtype detection, NGS was largely in agreement with Sanger sequencing but showed higher sensitivity for mixed subtype colonization within one host. This fact together with use of the combination of qPCR and NGS and obtaining information on the fecal protist load will be beneficial for epidemiological and surveillance studies.
- MeSH
- Blastocystis * genetika MeSH
- blastocystóza * diagnóza epidemiologie MeSH
- feces MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- molekulární patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- Blastocystis * genetika MeSH
- Dientamoeba genetika MeSH
- feces MeSH
- lidé MeSH
- syndrom dráždivého tračníku * MeSH
- zmrazování MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- dopisy MeSH
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.
- MeSH
- asymptomatické infekce epidemiologie MeSH
- Blastocystis klasifikace genetika izolace a purifikace MeSH
- blastocystóza epidemiologie parazitologie MeSH
- dítě MeSH
- feces parazitologie MeSH
- genetická variace MeSH
- lidé MeSH
- mladiství MeSH
- prevalence MeSH
- protozoální DNA genetika MeSH
- ribozomální DNA genetika MeSH
- střevní mikroflóra genetika MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Ázerbájdžán MeSH
- Československo MeSH
- Jordánsko MeSH
- Nigérie MeSH
- Súdán MeSH
- Tanzanie MeSH
Blastocystis sp. is a common intestinal protist colonizing the human intestine the prevalence of which varies across non-industrialized and industrialized countries. Its role in the human gut ecosystem remains unclear due to persisting gaps in knowledge of epidemiology and factors affecting gut colonization. Here, we aimed to expand the knowledge of the epidemiology of Blastocystis sp. in the gut-healthy humans in one of the industrialized European countries, including the distribution of its subtypes, the correlation between its occurrence and several factors such as lifestyle, contact with animals, age, and sex. A total of 288 stool samples were obtained from asymptomatic individuals over the entire age-range and 136 samples from animals with which the volunteers were in frequent contact. All samples were examined in parallel by PCR and xenic in vitro culture. Blastocystis sp. was detected in samples from both human and non-human hosts. In humans, the overall prevalence was 24% and eight subtypes were found; in animals, the prevalence was 10%, and only five subtypes were detected. A higher incidence of Blastocystis sp. was observed in individuals (i) traveling outside Europe, (ii) in frequent contact with livestock, and (iii) over 50 years of age. We found no effect on gender on Blastocystis sp. colonization.Summary: This study provides data on the prevalence and diversity of the gut protist Blastocystis sp. and its subtypes in a gut-healthy human population with emphasis on several factors such as contact with animals, lifestyle, age, and gender.
- MeSH
- Blastocystis * genetika MeSH
- blastocystóza * epidemiologie MeSH
- ekosystém MeSH
- feces MeSH
- lidé MeSH
- prevalence MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Evropa MeSH
The establishment of the mitochondrion is seen as a transformational step in the origin of eukaryotes. With the mitochondrion came bioenergetic freedom to explore novel evolutionary space leading to the eukaryotic radiation known today. The tight integration of the bacterial endosymbiont with its archaeal host was accompanied by a massive endosymbiotic gene transfer resulting in a small mitochondrial genome which is just a ghost of the original incoming bacterial genome. This endosymbiotic gene transfer resulted in the loss of many genes, both from the bacterial symbiont as well the archaeal host. Loss of genes encoding redundant functions resulted in a replacement of the bulk of the host's metabolism for those originating from the endosymbiont. Glycolysis is one such metabolic pathway in which the original archaeal enzymes have been replaced by bacterial enzymes from the endosymbiont. Glycolysis is a major catabolic pathway that provides cellular energy from the breakdown of glucose. The glycolytic pathway of eukaryotes appears to be bacterial in origin, and in well-studied model eukaryotes it takes place in the cytosol. In contrast, here we demonstrate that the latter stages of glycolysis take place in the mitochondria of stramenopiles, a diverse and ecologically important lineage of eukaryotes. Although our work is based on a limited sample of stramenopiles, it leaves open the possibility that the mitochondrial targeting of glycolytic enzymes in stramenopiles might represent the ancestral state for eukaryotes.
- MeSH
- biologická evoluce MeSH
- Blastocystis cytologie enzymologie genetika metabolismus MeSH
- energetický metabolismus MeSH
- genom mitochondriální MeSH
- glykolýza * MeSH
- mitochondrie genetika metabolismus MeSH
- rozsivky cytologie enzymologie genetika metabolismus MeSH
- symbióza MeSH
- transformace genetická MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Blastocystis is a common inhabitant of the human gut, colonizing at least one billion people at a prevalence ranging from <10% to 100% in healthy human populations globally. The majority of carriers remain asymptomatic, suggesting that Blastocystis is largely a commensal, though Blastocystis has also been implicated in disease in some people. However, there are no in vivo model systems in which to experimentally test the impact of Blastocystis on mammalian hosts and the gut ecosystem and determine which factors underlie these variable clinical outcomes. We evaluated a rat model for sustaining of a human-derived Blastocystis ST1 and assess colonization success and longevity. Because of the broad host range of Blastocystis, we compared the rat with three other rodent species to establish the reproducibility of our method. Blastocystis was introduced by esophageal gavage and colonization success evaluated by Blastocystis culture. Culture was also used to determine that all animals were negative prior to colonization and negative controls remain Blastocystis-free. In this study, Blastocystis ST1 established in 100% of the outbred rats (Rattus norvegicus) and gerbils (Meriones unguiculatus) challenged. Rats were colonized asymptomatically for more than one year, but Blastocystis ST1 was not transmitted between rats. Mus musculus strain CD1 and Mastomys coucha were not susceptible to Blastocystis ST1. Thus, rats appear to be a suitable in vivo model for studies of Blastocystis ST1, as do gerbils though testing was less extensive. This work lays the foundation for experimental work on the role of Blastocystis in health and disease.
- MeSH
- Blastocystis růst a vývoj patogenita MeSH
- blastocystóza diagnóza parazitologie MeSH
- centrifugace - gradient hustoty MeSH
- feces parazitologie MeSH
- Gerbillinae MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- modely nemocí na zvířatech * MeSH
- Murinae MeSH
- myši MeSH
- náchylnost k nemoci MeSH
- organismy bez specifických patogenů MeSH
- potkani Wistar MeSH
- zdravotní stav MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- srovnávací studie MeSH
