[The study of microbiota in patients with bullous lesions of the oral mucosa]. / Izuchenie mikrobioty slizistoi obolochki rta u patsientov s bulleznymi porazheniyami.

THE AIM OF THE STUDY: The study by the method of tissue polymerase chain reaction of the species composition of the microbiota of lesions of the oral mucosa in patients with bullous lesions. MATERIAL AND METHODS: Biopsy specimens of the oral mucosa of 51 patients were studied by the polymerase chain reaction method, of which 14 patients with pemphigus vulgaris, 17 patients with pemphigoid bullosa, and 20 patients with the bullous form of ruber lichen planus. 4 types of microorganisms have been identified - Fusobacterium, Streptococcus pneumoniae, Candida albicans, Ureaplasma spp. and viruses - Human Papillomavirus 16, Epstein-Barr virus and Citomegalovirus. RESULTS: In the study of the microbiota of bullous lesions, associations of microorganisms and viruses were established in a significant number of cases. Associations of Str.pneumoniae and C. albicans were quite common in patients with pemphigus vulgaris in 26.3%, pemphigoid bullosa in 20.0%, and in patients with the bullous form of ruber lichen planus in 14.3% of cases. In patients with pemphigus vulgaris, the association of Str.pneumoniae, C. albicans and EBV was noted in 31.6% of cases. In patients with the bullous form of ruber lichen planus in a high percentage of cases (28.6%), the associations of Str. pneumoniae, EBV and CMV. CONCLUSION: Identification at earlier stages of management of patients with bullous lesions Str. pneumoniae, Candida albicans, and Fusobacterium associated with herpes viruses should be regarded as one of the triggering mechanisms of an autoimmune conflict, which subsequently causes a specific clinical picture of these diseases.

Porphyromonas spp., Fusobacterium spp., and Bacteroides spp. dominate microbiota in the course of macropod progressive periodontal disease.

Macropod progressive periodontal disease (MPPD) is a necrotizing, polymicrobial, inflammatory disease commonly diagnosed in captive macropods. MPPD is characterized by gingivitis associated with dental plaque formation, which progresses to periodontitis and then to osteomyelitis of the mandible or maxilla. However, the underlying microbial causes of this disease remain poorly understood. In this study, we collected 27 oral plaque samples and associated clinical records from 22 captive Macropodidae and Potoroidae individuals that were undergoing clinical examination at Adelaide and Monarto Zoos in South Australia (15 healthy, 7 gingivitis and 5 periodontitis-osteomyelitis samples). The V3-V4 region of the 16S ribosomal RNA gene was sequenced using an Illumina Miseq to explore links between MPPD and oral bacteria in these animals. Compositional differences were detected between the microbiota of periodontitis-osteomyelitis cases compared to healthy samples (p-value with Bonferroni correction < 0.01), as well as gingivitis cases compared to healthy samples (p-value with Bonferroni correction < 0.05) using Permutational Multivariate Analysis of Variance (PERMANOVA). An overabundance of Porphyromonas, Fusobacterium, and Bacteroides taxa was also identified in animals with MPPD compared to healthy individuals using linear discriminant analysis effect size (LEfSe; p = < 0.05). An increased abundance of Desulfomicrobium also was detected in MPPD samples (LEfSe; p < 0.05), which could potentially reflect differences in disease progression. This is the first microbiota analysis of MPPD in captive macropods, and these results support a polymicrobial pathogenesis of MPPD, suggesting that the microbial interactions underpinning MPPD may be more complex than previously documented.

The Sulfur Microbial Diet and Risk of Colorectal Cancer by Molecular Subtypes and Intratumoral Microbial Species in Adult Men.

INTRODUCTION: We recently described the sulfur microbial diet, a pattern of intake associated with increased gut sulfur-metabolizing bacteria and incidence of distal colorectal cancer (CRC). We assessed whether this risk differed by CRC molecular subtypes or presence of intratumoral microbes involved in CRC pathogenesis (Fusobacterium nucleatum and Bifidobacterium spp.). METHODS: We performed Cox proportional hazards modeling to examine the association between the sulfur microbial diet and incidence of overall and distal CRC by molecular and microbial subtype in the Health Professionals Follow-Up Study (1986-2012). RESULTS: We documented 1,264 incident CRC cases among 48,246 men, approximately 40% of whom had available tissue data. After accounting for multiple hypothesis testing, the relationship between the sulfur microbial diet and CRC incidence did not differ by subtype. However, there was a suggestion of an association by prostaglandin synthase 2 (PTGS2) status with a multivariable adjusted hazard ratio for highest vs lowest tertile of sulfur microbial diet scores of 1.31 (95% confidence interval: 0.99-1.74, Ptrend = 0.07, Pheterogeneity = 0.04) for PTGS2-high CRC. The association of the sulfur microbial diet with distal CRC seemed to differ by the presence of intratumoral Bifidobacterium spp. with an adjusted hazard ratio for highest vs lowest tertile of sulfur microbial diet scores of 1.65 (95% confidence interval: 1.14-2.39, Ptrend = 0.01, Pheterogeneity = 0.03) for Bifidobacterium-negative distal CRC. We observed no apparent heterogeneity by other tested molecular markers. DISCUSSION: Greater long-term adherence to the sulfur microbial diet could be associated with PTGS2-high and Bifidobacterium-negative distal CRC in men. Additional studies are needed to further characterize the role of gut microbial sulfur metabolism and CRC.

Integration of constraint-based modeling with fecal metabolomics reveals large deleterious effects of Fusobacterium spp. on community butyrate production.

Characterizing the metabolic functions of the gut microbiome in health and disease is pivotal for translating alterations in microbial composition into clinical insights. Two major analysis paradigms have been used to explore the metabolic functions of the microbiome but not systematically integrated with each other: statistical screening approaches, such as metabolome-microbiome association studies, and computational approaches, such as constraint-based metabolic modeling. To combine the strengths of the two analysis paradigms, we herein introduce a set of theoretical concepts allowing for the population statistical treatment of constraint-based microbial community models. To demonstrate the utility of the theoretical framework, we applied it to a public metagenomic dataset consisting of 365 colorectal cancer (CRC) cases and 251 healthy controls, shining a light on the metabolic role of Fusobacterium spp. in CRC. We found that (1) glutarate production capability was significantly enriched in CRC microbiomes and mechanistically linked to lysine fermentation in Fusobacterium spp., (2) acetate and butyrate production potentials were lowered in CRC, and (3) Fusobacterium spp. presence had large negative ecological effects on community butyrate production in CRC cases and healthy controls. Validating the model predictions against fecal metabolomics, the in silico frameworks correctly predicted in vivo species metabolite correlations with high accuracy. In conclusion, highlighting the value of combining statistical association studies with in silico modeling, this study provides insights into the metabolic role of Fusobacterium spp. in the gut, while providing a proof of concept for the validity of constraint-based microbial community modeling.

Comprehensive profiling of the gut microbiota in patients with chronic obstructive pulmonary disease of varying severity.

Chronic obstructive pulmonary disease (COPD) is a chronic respiratory disease that reduces lung and respiratory function, with a high mortality rate. Severe and acute deterioration of COPD can easily lead to respiratory failure, resulting in personal, social, and medical burden. Recent studies have shown a high correlation between the gut microbiota and lung inflammation. In this study, we investigated the relationship between gut microbiota and COPD severity. A total of 60 COPD patients with varying severity according to GOLD guidelines were enrolled in this study. DNA was extracted from patients' stool and 16S rRNA data analysis conducted using high-throughput sequencing followed by bioinformatics analysis. The richness of the gut microbiota was not associated with COPD severity. The gut microbiome is more similar in stage 1 and 2 COPD than stage 3+4 COPD. Fusobacterium and Aerococcus were more abundant in stage 3+4 COPD. Ruminococcaceae NK4A214 group and Lachnoclostridium were less abundant in stage 2-4, and Tyzzerella 4 and Dialister were less abundant in stage 1. However, the abundance of a Bacteroides was associated with blood eosinophils and lung function. This study suggests that no distinctive gut microbiota pattern is associated with the severity of COPD. The gut microbiome could affect COPD by gut inflammation shaping the host immune system.

Colonic Adenocarcinoma Presenting as Splenic Abscess Secondary to Suspected Microperforation.

Splenic abscesses are a rare infection that usually requires seeding from another primary source; however, direct contact of bacteria can occur with microperforation secondary to colon cancer leading to abscess formation. This occurrence is rare, and through literature review only 12 previous cases have been reported with associated bacteremia. Our patient is a 62-year-old female who presented with left upper quadrant pain with a history of tobacco and alcohol abuse that was febrile and hypoxic. Blood cultures were obtained that eventually grew Fusobacterium mortiferum. Computed tomography of the abdomen and the pelvis revealed 2 splenic abscesses that were cultured to grow Escherichia coli and ß-hemolytic Streptococcus group C. Colonoscopy was performed, which identified 2 masses that were biopsied, and histopathology confirmed well-differentiated adenocarcinoma with possible muscular invasion. The patient had no other identifiable risk factors for bacterial seeding from another primary source. We present the first reported case report of splenic abscess secondary to colonic adenocarcinoma suspected microperforation associated with Fusobacterium mortiferum bacteremia.

Glycan cross-feeding supports mutualism between Fusobacterium and the vaginal microbiota.

Women with bacterial vaginosis (BV), an imbalance of the vaginal microbiome, are more likely to be colonized by potential pathogens such as Fusobacterium nucleatum, a bacterium linked with intrauterine infection and preterm birth. However, the conditions and mechanisms supporting pathogen colonization during vaginal dysbiosis remain obscure. We demonstrate that sialidase activity, a diagnostic feature of BV, promoted F. nucleatum foraging and growth on mammalian sialoglycans, a nutrient resource that was otherwise inaccessible because of the lack of endogenous F. nucleatum sialidase. In mice with sialidase-producing vaginal microbiotas, mutant F. nucleatum unable to consume sialic acids was impaired in vaginal colonization. These experiments in mice also led to the discovery that F. nucleatum may also "give back" to the community by reinforcing sialidase activity, a biochemical feature of human dysbiosis. Using human vaginal bacterial communities, we show that F. nucleatum supported robust outgrowth of Gardnerella vaginalis, a major sialidase producer and one of the most abundant organisms in BV. These results illustrate that mutually beneficial relationships between vaginal bacteria support pathogen colonization and may help maintain features of dysbiosis. These findings challenge the simplistic dogma that the mere absence of "healthy" lactobacilli is the sole mechanism that creates a permissive environment for pathogens during vaginal dysbiosis. Given the ubiquity of F. nucleatum in the human mouth, these studies also suggest a possible mechanism underlying links between vaginal dysbiosis and oral sex.

An oral health optimized diet reduces the load of potential cariogenic and periodontal bacterial species in the supragingival oral plaque: A randomized controlled pilot study.

This study aimed to investigate the effects of an oral health optimized diet on the composition of the supragingival oral plaque in a randomized controlled trial. Participants of the standard diet group (n = 5) had a diet high in processed carbohydrates and did not change their dietary behavior during the observation. The healthy diet group (n = 9) had to change the diet after 2 weeks from a diet high in processed carbohydrates to a diet low in carbohydrates, rich in omega-3 fatty acids, rich in vitamins C and D, antioxidants and fiber for 4 weeks. Saliva and supragingival plaque samples were taken at the end of week two and eight of the observation period to investigate the composition of microbiota in saliva and supragingival plaque. Data were subjected to an exploratory analysis to identify significant differences. Statistically significant differences were only found in the healthy diet group between the baseline (week 2) and the final sample (week 8) for specific species in plaque and saliva samples. A reduction of the total counts of Streptococcus mitis group, Granulicatella adiacens, Actinomyces spp., and Fusobacterium spp. was found in plaque samples of the healthy diet group. In saliva samples of the healthy diet group, the total counts of Actinomyces spp. and Capnocytophaga spp. decreased. A diet low in carbohydrates, rich in omega-3 fatty acids, rich in vitamins C and D, and rich in fiber reduced Streptococcus mitis group, Granulicatella adiacens, Actinomyces spp., and Fusobacterium spp. in the supragingival plaque.

Southern Chinese populations harbour non-nucleatum Fusobacteria possessing homologues of the colorectal cancer-associated FadA virulence factor.

OBJECTIVE: Fusobacteria are not common nor relatively abundant in non-colorectal cancer (CRC) populations, however, we identified multiple Fusobacterium taxa nearly absent in western and rural populations to be comparatively more prevalent and relatively abundant in southern Chinese populations. We investigated whether these represented known or novel lineages in the Fusobacterium genus, and assessed their genomes for features implicated in development of cancer. METHODS: Prevalence and relative abundances of fusobacterial species were calculated from 3157 CRC and non-CRC gut metagenomes representing 16 populations from various biogeographies. Microbial genomes were assembled and compared with existing reference genomes to assess novel fusobacterial diversity. Phylogenetic distribution of virulence genes implicated in CRC was investigated. RESULTS: Irrespective of CRC disease status, southern Chinese populations harboured increased prevalence (maximum 39% vs 7%) and relative abundances (average 0.4% vs 0.04% of gut community) of multiple recognised and novel fusobacterial taxa phylogenetically distinct from Fusobacterium nucleatum. Genomes assembled from southern Chinese gut metagenomes increased existing fusobacterial diversity by 14.3%. Homologues of the FadA adhesin linked to CRC were consistently detected in several monophyletic lineages sister to and inclusive of F. varium and F. ulcerans, but not F. mortiferum. We also detected increased prevalence and relative abundances of F. varium in CRC compared with non-CRC cohorts, which together with distribution of FadA homologues supports a possible association with gut disease. CONCLUSION: The proportion of fusobacteria in guts of southern Chinese populations are higher compared with several western and rural populations in line with the notion of environment/biogeography driving human gut microbiome composition. Several non-nucleatum taxa possess FadA homologues and were enriched in CRC cohorts; whether this imposes a risk in developing CRC and other gut diseases deserves further investigation.

Postpartum Fusobacterium gonidiaformans bacteremia.

We present a case of a healthy 29 year-old female with an uneventful vaginal delivery who had transient, sudden onset of rigors and fever 36 hours postpartum. She was found to have Fusobacterium gonidiaformans bacteremia due to retained placental tissue. We report this organism as it is not well-described and rarely reported. It does bear some similarities to other Fusobacterium species that have been reported to cause septicemia in young otherwise healthy patients.

Detection of Fusobacterium in oral and head and neck cancer samples: A systematic review and meta-analysis.

AIMS: This systematic review aimed to analyse: a) the presence and the abundance of Fusobacterium; b) the Fusobacterium species most often found, and c) the most common methods used for their identification in oral/head and neck cancer samples. DESIGN: A protocol was registered on PROSPERO database. This review was conducted following PRISMA guidelines. Literature search was performed on five electronic biomedical databases, namely Pubmed, Scopus, Web of Science, Embase, and Cochrane from their start dates to 30 August 2018. Two reviewers independently assessed the eligibility for inclusion; extracted the data; and evaluated the risk of bias. RESULTS: From 118 unique abstract records, 88 full-text articles were assessed for eligibility. According to inclusion and exclusion criteria, 17 publications were included in this review. Meta-analysis showed an increased prevalence of 6 % (95 % CI, 3-9) of Fusobacterium in tumour lesions than in non-tumour lesions (Fusobacterium prevalence of 16 % in tumour lesions and of 10 % in non-tumour lesions), and a 2.93 higher chance of Fusobacterium being present in tumour lesions (95 % CI, 1.47-5.81). The most common detection methods were based on molecular evidence (64.70 %) (95 % CI, 37.7-84.7). F. nucleatum was the most prevalent species (47.06 %) (95 % CI, 23.5-72). CONCLUSION: In conclusion, Fusobacterium is present and in higher abundance in oral/head and neck cancer samples when compared to non-cancer samples, suggesting that Fusobacterium may contribute to oral/head and neck cancer development.

Microbiota profile is different for early and invasive colorectal cancer and is consistent throughout the colon.

BACKGROUND AND AIM: Microbiota have been associated with several diseases including colorectal cancer (CRC). This study aimed to evaluate the microbiota in early/invasive CRC utilizing stool and cytological brushes to determine differences in relative abundance (RA). METHODS: Colonoscopy patients referred for endoscopic submucosal dissection or previous to CRC surgery were prospectively enrolled. Stool was collected pre-bowel preparation; and brush samples were taken during colonoscopy (three regions). DNA extraction, 16S rRNA next generation sequencing, and biostatistics (qiime and stamp software packages) followed. Primary outcome was the difference in RA of the Fusobacterium genus between the groups. Secondary outcomes included analyses of other microbiota. RESULTS: Twenty-five patients were included, of which 14 had invasive cancer (≥ 1000 mm into the submucosa). The three major genera for invasive cancer were Bacterioides, Oribacterium, and Fusobacterium, whereas for early cancer were Oribacterium, Bacterioides, and Prevotella (decreasing order of RA). There was a significantly higher RA of Fusobacterium in the invasive cancer group (9.65% vs 0.95%, respectively, P < 0.001). The RA of all genera was similar throughout the colon. In addition to Fusobacterium, the genera Corynebacterium, Enterococcus, Neisseria, Porphyromonas, and Sclegelella showed statistically higher RA in the invasive cancer group. Conversely, the genera Oribacterium, Desulfovibrio, Clostridiales, and Lactobacillus showed lower RA in the invasive cancer group. CONCLUSIONS: The RA of Fusobacterium is higher with invasive CRC than in early CRC patients. In addition, five other bacteria genera were found to be increased, and four decreased in invasive CRC patients. The microbiota per patient was similar throughout the colon.

Measuring the subgingival microbiota in periodontitis patients: Comparison of the surface layer and the underlying layers.

Periodontitis is a major cause of tooth loss in adults that initially results from dental plaque. Subgingival plaque pathogenesis is affected by both community composition and plaque structures, although limited data are available concerning the latter. To bridge this knowledge gap, subgingival plaques were obtained using filter paper (the fourth layer) and curette (the first-third layers) sequentially and the phylogenetic differences between the first-third layers and the fourth layer were characterized by sequencing the V3-V4 regions of 16S rRNA. A total of 11 phyla, 148 genera, and 308 species were obtained by bioinformatic analysis, and no significant differences between the operational taxonomic unit numbers were observed for these groups. In both groups, the most abundant species were Porphyromonas gingivalis and Fusobacterium nucleatum. Actinomyces naeslundii, Streptococcus intermedius, and Prevotella intermedia possessed relatively high proportions in the first-third layers; while in the fourth layer, both traditional pathogens (Treponema denticola and Campylobacter rectus) and novel pathobionts (Eubacterium saphenum, Filifactor alocis, Treponema sp. HOT238) were prominent. Network analysis showed that either of them exhibited a scale-free property and was constructed by two negatively correlated components (the pathogen component and the nonpathogen component), while the synergy in the nonpathogen component was lower in the first-third layers than that in the fourth layer. After merging these two parts into a whole plaque group, the negative/positive correlation ratio increased. With potential connections, the first-third layers and the fourth layer showed characteristic key nodes in bacterial networks.

Recovery of aerobic and anaerobic bacteria from patients with acute appendicitis using blood culture bottles / Recuperación de bacterias aerobias y anaerobias de pacientes con apendicitis aguda mediante botellas de hemocultivo

Introduction: Acute appendicitis is the first cause of acute abdomen, however, there is a little information about the associated bacteria and its sensibility profile. Objetive: To identify and to determine the resistance pattern of aerobic and anaerobic bacteria isolated in periapendicular fluid cultures taken in patients with acute appendicitis and to establish the proportions of isolates according to the clinical phase. Materials and methods: A descriptive and prospective study was undertaken at the Hospital Universitario de San José (Bogotá, Colombia) of patients older than sixteen years of age, undergoing an open appendectomy. A sample of periappendiceal fluid was taken, which was deposited directly into aerobic and anaerobic blood culture bottles. Results: One hundred and fifty-four patients were included. The overall positivity of cultures was 87% (n=1344); 77% (n=118) for aerobes and 51% (n=79) for anaerobes. The proportion of positive cultures was lower in the uncomplicated appendicitis cases as compared to the complicated ones (80% (66/83) vs. 95%(67/71), p = 0.003). The microorganisms isolated most frequently were: Escherichia coli (53%) (n=84); Bacteroides spp. (25%) (n=25); Propionibacterium acnes (21%) (n=21); coagulase negative Staphylococci (17%) (n=27); Enterococcus spp. (11%) (n=15), and Fusobacterium spp. (11%) (n=11). The sensitivity of E. coli to ampicillin/sulbactam was 30%. The sensitivity of Bacteroides spp. to clindamycin and ampicillin/sulbactam was 91%. All anaerobe isolates were sensitive to piperacillin/tazobactam, ertapenem, meropenem and metronidazole. Conclusions: Intraoperative cultures in acute appendicits are relevant in order to determine the local epidemiological pattern and to establish prophylactic and therapeutic antibiotics for this pathology; direct inoculation in blood culture bottles allows a high recovery of microorganisms.

Introduccción. La apendicitis aguda es la primera causa de abdomen agudo; sin embargo, poco se conoce sobre las bacterias asociadas y su perfil de sensibilidad. Objetivo. Identificar y determinar el patrón de resistencia de las bacterias aerobias y anaerobias aisladas en cultivo de líquido periapendicular tomado de los pacientes con apendicitis aguda, y establecer la proporción de bacterias según la fase clínica. Materiales y métodos. Se llevó a cabo un estudio descriptivo y prospectivo en el Hospital Universitario de San José de Bogotá (Colombia), en pacientes mayores de 16 años sometidos a apendicectomía abierta. Se tomaron muestras de líquido periapendicular, las cuales se sembraron directamente en botellas de hemocultivos para aerobios y anaerobios. Resultados. Se incluyeron 154 pacientes. Del total de cultivos, el 87 % (n=134) fueron positivos: 77 % (n=118) para aerobios y 51 % (n=79) para anaerobios. La proporción de cultivos positivos fue inferior en los casos de apendicitis no complicada, en comparación con aquellos de apendicitis complicada (80 % (66/83) Vs. 95 % (67/71); p=0,003). Los microorganismos aislados con mayor frecuencia fueron: Escherichia coli (53 %) (n=84), Bacteroides sp. (25 %) (n=25), Propionibacterium acnes (21 %) (n=21), Staphylococci coagulasa negativo (17 %) (n=27), Enterococcus sp. (10 %) (n=15) y Fusobacterium sp. (11 %) (n=11). La sensibilidad de E. coli a la amplicilina sulbactam fue de 30 %. La sensibilidad de Bacteroides spp. a la clindamicina y la ampicilina sulbactam fue de 91 %. El 100 % de los anaerobios fueron sensibles a piperacilina tazobactam, ertapenem, meropenem y metronidazol. Conclusiones. Los cultivos intraoperatorios son pertinentes en la apendicitis para determinar el patrón epidemiológico local, y establecer los antibióticos profilácticos y terapéuticos para esta enfermedad. Su siembra directa en botellas de hemocultivo permite una gran recuperación de microorganismos.

The Role of the Gut Microbiota in Colorectal Cancer Causation.

Here, we reviewed emerging evidence on the role of the microbial community in colorectal carcinogenesis. A healthy gut microbiota promotes intestinal homeostasis and can exert anti-cancer effects; however, this microbiota also produces a variety of metabolites that are genotoxic and which can negatively influence epithelial cell behaviour. Disturbances in the normal microbial balance, known as dysbiosis, are frequently observed in colorectal cancer (CRC) patients. Microbial species linked to CRC include certain strains of Bacteroides fragilis, Escherichia coli, Streptococcus gallolyticus, Enterococcus faecalis and Fusobacterium nucleatum, amongst others. Whether these microbes are merely passive dwellers exploiting the tumour environment, or rather, active protagonists in the carcinogenic process is the subject of much research. The incidence of chemically-induced tumours in mice models varies, depending upon the presence or absence of these microorganisms, thus strongly suggesting influences on disease causation. Putative mechanistic explanations differentially link these strains to DNA damage, inflammation, aberrant cell behaviour and immune suppression. In the future, modulating the composition and metabolic activity of this microbial community may have a role in prevention and therapy.

Specific increase of Fusobacterium in the faecal microbiota of neonatal calves infected with Cryptosporidium parvum.

The faecal microbiota plays a critical role in host health, with alterations in the human faecal microbial composition associated with various conditions, particularly diarrhoeal diseases. However, little is known about microbial changes during cryptosporidiosis, one of the most important diarrhoeal diseases caused by protozoa in cattle. In this study, alterations in the faecal microbiota of neonatal calves as a result of Cryptosporidium parvum infection were investigated on a C. parvum-positive farm. Comparisons were made among groups of C. parvum-infected, rotavirus-infected, and the pathogen-negative calves. A specific increase in the abundance of Fusobacterium was observed in the faecal microbiota of C. parvum-infected animals. Diarrhoea severity increased in accordance with the abundance of C. parvum and Fusobacterium. Moreover, the specific increase of Fusobacterium appeared to be a universal feature of C. parvum infection, since neonatal calves from geographically separated areas showed the same result. These observations indicated that the growth of Fusobacterium may be an important aggravating factor of cryptosporidiosis.

Quantitative microbiome profiling disentangles inflammation- and bile duct obstruction-associated microbiota alterations across PSC/IBD diagnoses.

Recent work has highlighted the importance of confounder control in microbiome association studies1,2. For instance, multiple pathologies previously linked to gut ecosystem dysbiosis display concomitant changes in stool consistency3-6, a major covariate of microbiome variation2,7. In those cases, observed microbiota alterations could largely reflect variation in faecal water content. Moreover, stool moisture variation has been linked to fluctuations in faecal microbial load, inducing artefacts in relative abundance profile analyses8,9. Hence, the identification of associations between the gut microbiota and specific disease manifestations in pathologies with complex aetiologies requires a deconfounded, quantitative assessment of microbiome variation. Here, we revisit a disease association microbiome data set comprising 106 patients with primary sclerosing cholangitis (PSC) and/or inflammatory bowel disease10. Assessing quantitative taxon abundances9, we study microbiome alterations beyond symptomatic stool moisture variation. We observe an increased prevalence of a low cell count Bacteroides 2 enterotype across the pathologies studied, with microbial loads correlating inversely with intestinal and systemic inflammation markers. Quantitative analyses allow us to differentiate between taxa associated with either intestinal inflammation severity (Fusobacterium) or cholangitis/biliary obstruction (Enterococcus) among previously suggested PSC marker genera. We identify and validate a near-exclusion pattern between the inflammation-associated Fusobacterium and Veillonella genera, with Fusobacterium detection being restricted to Crohn's disease and patients with PSC-Crohn's disease. Overall, through absolute quantification and confounder control, we single out clear-cut microbiome markers associated with pathophysiological manifestations and disease diagnosis.