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Abstract：
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.
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.
摘要：
背景：植物乳杆菌HEAL9和副干酪乳杆菌8700:2在补充6个月后对乳糜泻自身免疫儿童的粪便细菌组有积极影响。本研究的目的是研究植物乳杆菌HEAL9和副干酪乳杆菌8700：2对单细胞寄生虫的影响，主要集中在胚泡。
方法：收集了78名患有乳糜泻自身免疫的瑞典儿童的粪便样本，双盲,安慰剂对照临床试验，接受植物乳杆菌HEAL9和副干酪乳杆菌8700:2（n=38）或安慰剂（n=40）的混合物补充。在基线和干预3个月和6个月后，共收集了227个粪便样本，分别,通过定量实时PCR和大规模平行扩增子测序对囊胚进行回顾性分析。通过非靶向18SrDNA扩增子测序检测其他单细胞寄生虫，并通过实时PCR进行验证。通过使用V3-V4区域的16SrDNA谱分析来表征寄生虫与细菌群落之间的关系。
结果：确定了三种不同的单细胞原生生物，其中脆弱的Dientamoeba患病率最高(23.1%，18/78儿童），其次是胚泡（15.4%，12/78）和Entamoebaspp。（2.6%，2/78).随着时间的推移，原生生物的数量是稳定的，并且不受益生菌干预的影响（对于囊胚，P=0.14，D.fragilis的P=0.10)。原生生物的积极性与细菌组多样性的增加有关(通过多个指数衡量，P<0.03)。细菌组成受原生生物的影响：囊胚的阳性与Akkermansia呈负相关（在属及其家族的水平上，订单,类和门)；P<0.002)，粪杆菌（P=0.003）和Romboutsia（P=0.029）；D.fragilis的阳性与肠杆菌科（P=0.016）和科氏杆菌科（P=0.022）和Flavonifractor属（P<0.001）呈负相关，粪杆菌(P=0.009),衣原体（P=0.029），Ruminococus(P<0.001)和颗粒菌(P=0.018)。
结论：在患有乳糜泻自身免疫的儿童中，单细胞原生生物的患病率较低。无论益生菌干预如何，定植都是稳定的，并且与粪便细菌组的多样性增加有关，但与一些有益细菌成反比。
方法：收集了78名患有乳糜泻自身免疫的瑞典儿童的粪便样本，双盲,安慰剂对照临床试验，接受植物乳杆菌HEAL9和副干酪乳杆菌8700:2（n=38）或安慰剂（n=40）的混合物补充。在基线和干预3个月和6个月后，共收集了227个粪便样本，分别,通过定量实时PCR和大规模平行扩增子测序对囊胚进行回顾性分析。通过非靶向18SrDNA扩增子测序检测其他单细胞寄生虫，并通过实时PCR进行验证。通过使用V3-V4区域的16SrDNA谱分析来表征寄生虫与细菌群落之间的关系。
结果：确定了三种不同的单细胞原生生物，其中脆弱的Dientamoeba患病率最高(23.1%，18/78儿童），其次是胚泡（15.4%，12/78）和Entamoebaspp。（2.6%，2/78).随着时间的推移，原生生物的数量是稳定的，并且不受益生菌干预的影响（对于囊胚，P=0.14，D.fragilis的P=0.10)。原生生物的积极性与细菌组多样性的增加有关(通过多个指数衡量，P<0.03)。细菌组成受原生生物的影响：囊胚的阳性与Akkermansia呈负相关（在属及其家族的水平上，订单,类和门)；P<0.002)，粪杆菌（P=0.003）和Romboutsia（P=0.029）；D.fragilis的阳性与肠杆菌科（P=0.016）和科氏杆菌科（P=0.022）和Flavonifractor属（P<0.001）呈负相关，粪杆菌(P=0.009),衣原体（P=0.029），Ruminococus(P<0.001)和颗粒菌(P=0.018)。
结论：在患有乳糜泻自身免疫的儿童中，单细胞原生生物的患病率较低。无论益生菌干预如何，定植都是稳定的，并且与粪便细菌组的多样性增加有关，但与一些有益细菌成反比。
