Sébastien Lacroix

Preliminary experiments were performed to assess the use of intestinal Bacteroides spp. as indicators of fecal contamination of water. Viable counts of Bacteroides fragilis, an anaerobic bacterium, declined more rapidly than those of Escherichia coli and Streptococcus faecalis. However, a fluorescent antiserum prepared against B. fragilis successfully detected high proportions (18 to greater than 50%) of B. fragilis cells suspended for 8 days in aerobic water in dialysis bags at the ambient temperature. These percentages were higher than the percent viable recoveries of the two indicator bacteria used for comparison. Thus, the fluorescent antiserum test for B. fragilis might serve as a useful indicator of fecal contamination of water. An advantage of this approach over coliform analysis is the rapidity at which the test can be performed.

The gut microbiota is a unique ecosystem of microorganisms interacting with the host through several biochemical mechanisms. The endocannabinoidome (eCBome), a complex signaling system including the endocannabinoid system, approximately 50 receptors and metabolic enzymes, and more than 20 lipid mediators with important physiopathologic functions, modulates gastrointestinal tract function and may mediate host cell-microbe communications there. Germ-free (GF) mice, which lack an intestinal microbiome and so differ drastically from conventionally raised (CR) mice, offer a unique opportunity to explore the eCBome in a microbe-free model and in the presence of a reintroduced functional gut microbiome through fecal microbiota transplant (FMT). We aimed to gain direct evidence for a link between the microbiome and eCBome systems by investigating eCBome alterations in the gut in GF mice before and after FMT. Basal eCBome gene expression and lipid profiles were measured in various segments of the intestine of GF and CR mice at juvenile and adult ages using targeted quantitative PCR transcriptomics and LC-MS/MS lipidomics. GF mice exhibited age-dependent modifications in intestinal eCBome gene expression and lipid mediator levels. FMT from CR donor mice to age-matched GF male mice reversed several of these alterations, particularly in the ileum and jejunum, after only 1 week, demonstrating that the gut microbiome directly impacts the host eCBome and providing a cause-effect relationship between the presence or absence of intestinal microbes and eCBome signaling. These results open the way to new studies investigating the mechanisms through which intestinal microorganisms exploit eCBome signaling to exert some of their physiopathologic functions.

ANITA™Mox is a Veolia process using moving-bed biofilm reactor (MBBR) technology tested and validated in full-scale for energy- and cost-effective autotrophic N-removal from sidestream effluent using anammox (ANaerobic AMMonium OXidation) bacteria. In order to increase the ANITA™Mox process performances under different operating conditions (e.g. mainstream and sidestream application), substrate transport and accessibility inside the biofilm must be enhanced. In this work, (i) two laboratory scale biofilm ANITA™Mox reactors were operated using different configurations (IFAS - integrated fixed-film activated sludge - and MBBR) and (ii) the distribution of the anammox (AnAOB) and ammonia-oxidizing bacteria (AOB) in the suspended sludge and the biofilm was characterized using molecular tools (qPCR). This study showed that in IFAS configuration, the ANITA™Mox process achieved very high N-removal rate (up to 8 gN/m².d), which was three to four times higher than that achieved in the pure MBBR mode. The high concentration of suspended solids (mixed liquor suspended solids (MLSS)) in the bulk obtained within the IFAS mode induces a very efficient bacterial distribution between the AOB and AnAOB population. AnAOB activity mainly occurs in the biofilm (96% of total AnAOB in the reactor), whereas nitritation by AOB mostly takes place in the suspended phase (93% of total AOB). This spatial distribution observed in the IFAS reactor results from a natural selection due to more easily substrate accessibility for AOB in the bulk (NH4(+), O2) creating higher nitrite concentration in the bulk liquid compare to pure MBBR mode. The efficient control of MLSS level in the IFAS reactor is a key parameter to enhance the nitrite production by AOB and increase the substrate availability in the AnAOB-enriched biofilm leading to higher N-removal rate. These promising results obtained at laboratory scale have been further confirmed in on-going full-scale IFAS ANITA™Mox trials opening new roads for the widespread application of a very compact and robust ANITA™Mox process for sidestream but also mainstream cost-effective N-removal.

When a high titre inhibitor develops in a patient with haemophilia, attempts are made to eradicate it through immune tolerance induction therapy (ITI) involving the frequent and regular administration of factor, usually for months to years. ITI is successful in only two thirds of patients prompting investigators to explore alternate regimens to use in haemophiliacs failing conventional ITI. Rituximab is an anti-CD20 monoclonal antibody, which has shown promise in the treatment of B-cell-mediated disorders. We developed a protocol for the use of rituximab in haemophilia A (HA) patients failing conventional ITI or in those haemophiliacs where the likelihood of success of conventional ITI is poor. Patients receive 375 mg m(-2) of intravenous rituximab weekly for 4 weeks followed by monthly (up to 5 months) until inhibitor disappearance and establishment of normal FVIII pharmacokinetics (recovery and half-life). Patients are concurrently placed on recombinant FVIII (100 U kg(-1) day(-1)). We have placed five haemophiliacs (four children with severe HA, and one adult with mild HA) on this protocol. In three patients (two with severe HA and one with mild HA) inhibitors disappeared although in neither severe haemophiliac did FVIII pharmacokinetics completely normalize. The fourth patient had a significant drop in inhibitor titres although not a complete disappearance of the inhibitor. All four of these patients ceased bleeding following rituximab. The fifth patient had no response to rituximab. This non-responding patient was not placed on concurrent FVIII. Our five cases suggest that rituximab may hold promise in the eradication of inhibitors. Prospective randomized studies are required to determine the value of this agent in inhibitor management.