Gut microbiota plays a dual role in chronic kidney disease (CKD) and is closely linked to production of uremic toxins. Strategies of reducing uremic toxins by targeting gut microbiota are emerging. It is known that Chinese medicine rhubarb enema can reduce uremic toxins and improve renal function. However, it remains unknown which ingredient or mechanism mediates its effect. Here we utilized a rat CKD model of 5/6 nephrectomy to evaluate the effect of emodin, a main ingredient of rhubarb, on gut microbiota and uremic toxins in CKD. Emodin was administered via colonic irrigation at 5ml (1mg/day) for four weeks. We found that emodin via colonic irrigation (ECI) altered levels of two important uremic toxins, urea and indoxyl sulfate (IS), and changed gut microbiota in rats with CKD. ECI remarkably reduced urea and IS and improved renal function. Pyrosequencing and Real-Time qPCR analyses revealed that ECI resumed the microbial balance from an abnormal status in CKD. We also demonstrated that ten genera were positively correlated with Urea while four genera exhibited the negative correlation. Moreover, three genera were positively correlated with IS. Therefore, emodin altered the gut microbiota structure. It reduced the number of harmful bacteria, such as Clostridium spp. that is positively correlated with both urea and IS, but augmented the number of beneficial bacteria, including Lactobacillus spp. that is negatively correlated with urea. Thus, changes in gut microbiota induced by emodin via colonic irrigation are closely associated with reduction in uremic toxins and mitigation of renal injury.

Rapid and accurate detection and identification of microbial pathogens causing urinary tract infections allow prompt and specific treatment. We optimized specimen processing to maximize the limit of detection (LOD) by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and evaluated the capacity of combination of MALDI-TOF MS and urine analysis (UA) for direct detection and identification of bacterial pathogens from urine samples. The optimal volumes of processed urine, formic acid/acetonitrile, and supernatant spotted onto the target plate were 15 ml, 3 l, and 3 l, respectively, yielding a LOD of 1.0 ϫ 10 5 CFU/ml. Among a total of 1,167 urine specimens collected from three hospital centers, 612 (52.4%) and 351 (30.1%) were, respectively, positive by UA and urine culture. Compared with a reference method comprised of urine culture and 16S rRNA gene sequencing, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of MALDI-TOF MS alone and MALDI-TOF MS coupled with UA were 86.6% versus 93.4% ( 2 ϭ 8.93; P Ͻ 0.01), 91.5% versus 96.3% ( 2 ϭ 7.06; P Ͻ 0.01), 81.5% versus 96.4% ( 2 ϭ 37.32; P Ͻ 0.01), and 94.1% versus 93.1% ( 2 ϭ 0.40; P Ͼ 0.05), respectively. No significant performance differences were revealed among the three sites, while specificity and NPV of MALDI-TOF MS for males were significantly higher than those for females (specificity, 94.3% versus 77.3%, 2 ϭ 44.90, P Ͻ 0.01; NPV, 95.5% versus 86.1%, 2 ϭ 18.85, P Ͻ 0.01). Our results indicated that the optimization of specimen processing significantly enhanced analytical sensitivity and that the combination of UA and MALDI-TOF MS provided an accurate and rapid detection and identification of bacterial pathogens directly from urine.KEYWORDS identification, urine, pathogen, MALDI-TOF MS, urine analysis U rinary tract infections (UTIs) are among the most common bacterial infections seen in women. The initial treatment of UTI is mostly empirical, and immediately initiating therapy avoids serious complications and shortens the time of patient discomfort (1, 2). Diagnosis of UTI is currently based on the following three criteria: (i) clinical symptoms, (ii) detection of signs of infection in the urine, and (iii) detection and

Five bacterial strains (SYSU YG23T, SYSU 10HL1970T, 10HP82-10, 10HL1938, 10HP457) isolated from water reservoirs of cooling systems were characterized using a polyphasic taxonomic approach. The isolates were Gram-stain-negative, strictly aerobic and non-motile. Growth was enhanced in the presence of l-cysteine. The major fatty acids (>5 %) for the five strains were C10 : 0, C16 : 0, C16 : 0 3-OH, C18 : 0 3-OH and C18 : 1ω9c. Ubiquinone-8 was detected as the respiratory quinone while the polar lipid profile consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, three unidentified phospholipids, two unidentified aminophospholipids and three unidentified glycolipids. The strains shared 16S rRNA gene sequence similarities of 99.0-99.2 % with Francisella guangzhouensis 08HL01032T but less than 95.2 % with other members of the family Francisellaceae. The phylogenetic dendrogram based on 16S rRNA gene sequences showed that these strains form a separate cluster along with Francisella guangzhouensis. This cluster was also confirmed from multilocus-sequence typing based on sequences of the mdhA, rpoB and sdhA genes. Matrix-assisted laser desorption ionization time-of-flight MS analyses of the strains along with closely and distantly related Francisella strains also showed a distinct cluster for these strains. Based on the findings from the polyphasic taxonomy studies, the strains were considered to represent two novel species of a new genus for which the names Allofrancisella inopinata gen. nov., sp. nov. (type strain SYSU YG23T=KCTC 42968T=DSM 101834T) and Allofrancisella frigidaquae sp. nov. (type strain SYSU 10HL1970T=KCTC 42969T=DSM 101835T) are proposed. In addition, Francisella guangzhouensisQu et al. 2013 is proposed to be transferred to this new genus as Allofrancisella guangzhouensis comb. nov.

N-3-(Oxododecanoyl)-L-homoserine lactone (C12) is a small bacterial signaling molecule secreted by Pseudomonas aeruginosa (PA), which activates mammalian cells through TLR4-independent mechanisms. C12 acts as an immunosuppressant and it has been shown to modulate murine bone marrow-derived dendritic cell-mediated T-helper 2 (Th2) cell polarizations in vitro. In the present study, we initially examined the impact of C12 on the maturation of human monocyte-derived dendritic cells

AFrancisella-like bacterium, designated strain SYSU HZH-2, was isolated from a water sample collected from Haizhu Lake, Guangzhou, China. The bacterium was fastidious, and required an exogenous source of l-cysteine for its growth on artificial media. Cells were Gram-stain-negative, coccobacilli, non-motile and non-spore-forming. The strain shared highest 16S rRNA gene sequence similarities with Cysteiniphilum litorale SYSU D3-2 (94.6 % identity), Fangia hongkongensis UST040201-002 (93.2 %) and Caedibacter taeniospiralis 51 (91.6 %). This strain possessed ubiquinone-8 as the respiratory quinone; diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol as the known polar lipids, and anteiso-C15 : 0 and C18 : 0 as the major fatty acids (>10 % of total fatty acids). The dendrograms based on 16S rRNA gene sequence analysis showed that it formed a separate cluster along with Cysteiniphilum litorale SYSU D3-2, Caedibactertaeiniospiralis 51 and Fangia hongkongensis UST040201-002. Based on the 16S rRNA gene sequence identity and differences in other phenotypic characteristics, the strain is considered to represent a novel species of a novel genus, for which the name Fastidiosibacter lacustris gen. nov., sp. nov. is proposed. The type strain of the type species Fastidiosibacter lacustris is SYSU HZH-2 (=NBRC 112274 = CGMCC 1.15950). Additionally, the new taxon along with the genera Caedibacter, Cysteiniphilum and Fangia (family unassigned) were distinctly separated from the related families Francisellaceae, Piscirickettsiaceae and Thiotrichaeae in the phylogenetic trees. Therefore, we proposed a new family Fastidiosibacteraceae fam. nov. within the order Thiotrichales to accommodate these four genera.