A Comprehensive Evaluation of Colonic Mucosal Isolates of Sutterella wadsworthensis from Inflammatory Bowel Disease

Abstract

Inflammatory bowel disease (IBD) arises in genetically susceptible individuals as a result of an unidentified environmental trigger, possibly a hitherto unknown bacterial pathogen. Twenty-six clinical isolates of Sutterella wadsworthensis were obtained from 134 adults and 61 pediatric patients undergoing colonoscopy, of whom 69 and 29 respectively had IBD. S. wadsworthensis was initially more frequently isolated from IBD subjects, hence this comprehensive study was undertaken to elucidate its role in IBD. Utilizing these samples, a newly designed PCR was developed, to study the prevalence of this bacterium in adult patients with ulcerative colitis (UC). Sutterella wadsworthensis was detected in 83.8% of adult patients with UC as opposed to 86.1% of control subjects (p = 0.64). Selected strains from IBD cases and controls were studied to elicit morphological, proteomic, genotypic and pathogenic differences. This study reports Scanning Electron Microscopy (SEM) appearances and characteristic MALDI-TOF MS protein profiles of S. wadsworthensis for the very first time. SEM showed that the bacterium is pleomorphic, existing in predominantly two morphological forms, long rods and coccobacilli. No differences were noted in the MALDI-TOF mass spectrometry proteomic analysis. There was no distinct clustering of strains identified from cases and controls on sequence analysis. Cytokine response after monocyte challenge with strains from patients with IBD and controls did not yield any significant differences. Our studies indicate that S. wadsworthensis is unlikely to play a role in the pathogenesis of IBD. Strains from cases of IBD could not be distinguished from those identified from controls.

Mucosal Prevalence and Interactions with the Epithelium Indicate Commensalism of Sutterella spp.

“…and S. wadsworthensis from the intestinal biopsy samples was carried out using previously described quantitative real-time PCR (qPCR) assays (Suzuki and Giovannoni, 1996; Paliy et al, 2009; Mukhopadhya et al, 2011; Williams et al, 2012). The standard DNA for Sutterella genus assay was prepared from amplified 260-bp region between V6 and V8 of the 16S rRNA gene from S. wadsworthensis (ATCC 51579).…”

“…Primers were SuttFor (5′-CGC GAA AAA CCT TAC CTA GCC-3′) and SuttRev (5′-GAC GTG TGA GGC CCT AGC C-3′) (Williams et al, 2012). For S. wadsworthensis specific assay standard, a 555-bp long PCR-fragment of the 16S rRNA gene from S. wadsworthensis was amplified using forward primer SWF (5′-GAC GAA AAG GGA TGC GAT AA-3′) and reverse primer SWR (5′-CTG GCA TGT CAA GGC TAG GT-3′) (Mukhopadhya et al, 2011). The standard DNA for universal assay was produced by amplifying a 310-bp long fragment of the 16S rRNA gene from Prevotella melaninogenica using universal bacterial primers 27F-DegL (5′-AGR GTT YGA TYM TGG CTC AG-3′) (Paliy et al, 2009) and 338R (5′-TGC TGC CTC CCG TAG GAG T-3′) (Suzuki and Giovannoni, 1996).…”

“…have been suspected to play a part in the pathogenesis of inflammatory bowel disease (IBD; Mangin et al, 2004; Gophna et al, 2006; Lavelle et al, 2015), in other studies no difference in the prevalence of Sutterella spp. has been found between the IBD patients and the healthy subjects (Mukhopadhya et al, 2011; Hansen et al, 2013). Furthermore, Sutterella spp.…”

“…The lack of knowledge is rather surprising since Sutterella spp. have been reported to be highly prevalent and abundant in the intestinal mucosa of humans (Mukhopadhya et al, 2011; Cheng et al, 2013; Hansen et al, 2013). Firstly, S. wadsworthensis has been detected in 86 and 71% of adults and children, respectively (Mukhopadhya et al, 2011; Hansen et al, 2013).…”

275 The Role of Curcumin in Modulating Colonic Microbiota During Colitis and Colon Cancer Prevention

“…40,62 Analysis with the focus on the most abundant KEGG orthologs, which were affected by curcuminsupplementation in AOM/Il10 −/− mice showed reduced abundance of functions involved in transport, transcription factors, bacterial chemotaxis and bacterial motility proteins. 56,57,61,[63][64][65][66][67] Curcumin has been observed to inhibit bacterial motility and cytotoxicity of Vibrio vulnificus 68 , to arrest Helicobacter pylori growth during infections 69 , as well as inhibit biofilm formation in Pseudomonas aeruginosa 70 . This suggests that the protective effect of 0.5% curcumin diet shifts the microbial composition to be more stationary, which may contribute to a decrease in microbial invasiveness and a decrease in overall disease severity.…”