Versatile and efficient mammalian genome editing with Type I-C CRISPR System of Desulfovibrio vulgaris.

CRISPR-Cas tools for mammalian genome editing typically rely on single Cas9 or Cas12a proteins. While type I CRISPR systems in Class I may offer greater specificity and versatility, they are not well-developed for genome editing. Here, we present an alternative type I-C CRISPR system from Desulfovibrio vulgaris (Dvu) for efficient and precise genome editing in mammalian cells and animals. We optimized the Dvu type I-C editing complex to generate precise deletions at multiple loci in various cell lines and pig primary fibroblast cells using a paired PAM-in crRNA strategy. These edited pig cells can serve as donors for generating transgenic cloned piglets. The Dvu type I-C editor also enabled precise large fragment replacements with homology-directed repair. Additionally, we adapted the Dvu-Cascade effector for cytosine and adenine base editing, developing Dvu-CBE and Dvu-ABE systems. These systems efficiently induced C-to-T and A-to-G substitutions in human genes without double-strand breaks. Off-target analysis confirmed the high specificity of the Dvu type I-C editor. Our findings demonstrate the Dvu type I-C editor's potential for diverse mammalian genome editing applications, including deletions, fragment replacement, and base editing, with high efficiency and specificity for biomedicine and agriculture.

Low dose rifaximin combined with N-acetylcysteine is superior to rifaximin alone in a rat model of IBS-D: a randomized trial.

Rifaximin is FDA-approved for treatment of irritable bowel syndrome with diarrhea (IBS-D), but poor solubility may limit its efficacy against microbes in the mucus layer, e.g. Escherichia coli. Here we evaluate adding the mucolytic N-acetylcysteine (NAC) to improve rifaximin efficacy. In a resazurin checkerboard assay, combining rifaximin with NAC had significant synergistic effects in reducing E. coli levels. The optimal rifaximin + NAC combination was then tested in a validated rat model of IBS-D (induced by cytolethal distending toxin [CdtB] inoculation). Rats were inoculated with vehicle and treated with placebo (Control-PBS) or rifaximin + NAC (Control-Rif + NAC, safety), or inoculated with CdtB and treated with placebo (CdtB-PBS), rifaximin (CdtB-Rifaximin), or rifaximin + NAC (CdtB-Rif + NAC) for 10 days. CdtB-inoculated rats (CdtB-PBS) developed wide variability in stool consistency (P = 0.0014) vs. controls (Control-PBS). Stool variability normalized in rats treated with rifaximin + NAC (CdtB-Rif + NAC) but not rifaximin alone (CdtB-Rifaximin). Small bowel bacterial levels were elevated in CdtB-PBS rats but normalized in CdtB-Rif + NAC but not CdtB-Rifaximin rats. E. coli and Desulfovibrio spp levels (each associated with different IBS-D microtypes) were also elevated in CdtB-inoculated (CdtB-PBS) but normalized in CdtB-Rif + NAC rats. Cytokine levels normalized only in CdtB-Rif + NAC rats, in a manner predicted to be associated with reduced diarrhea driven by reduced E. coli. These findings suggest that combining rifaximin with NAC may improve the percentage of IBS-D patients responding to treatment.

Magnesium Oxide Reduces Anxiety-like Behavior in Mice by Inhibiting Sulfate-Reducing Bacteria.

The gut microbiota-brain axis allows for bidirectional communication between the microbes in our gastrointestinal (GI) tract and the central nervous system. Psychological stress has been known to disrupt the gut microbiome (dysbiosis) leading to anxiety-like behavior. Pathogens administered into the gut have been reported to cause anxiety. Whether commensal bacteria affect the gut-brain axis is not well understood. In this study, we examined the impact of a commensal sulfate-reducing bacteria (SRB) and its metabolite, hydrogen sulfide (H2S), on anxiety-like behavior. We found that mice gavaged with SRB had increased anxiety-like behavior as measured by the open field test. We also tested the effects of magnesium oxide (MgO) on SRB growth both in vitro and in vivo using a water avoidance stress (WAS) model. We found that MgO inhibited SRB growth and H2S production in a dose-dependent fashion. Mice that underwent psychological stress using the WAS model were observed to have an overgrowth (bloom) of SRB (Deferribacterota) and increased anxiety-like behavior. However, WAS-induced overgrowth of SRB and anxiety-like behavioral effects were attenuated in animals fed a MgO-enriched diet. These findings supported a potential MgO-reversible relationship between WAS-induced SRB blooms and anxiety-like behavior.

Desulfovibrio vulgaris caused gut inflammation and aggravated DSS-induced colitis in C57BL/6 mice model.

BACKGROUND: Sulfate-reducing bacteria (SRB) is a potential pathogen usually detected in patients with gastrointestinal diseases. Hydrogen sulfide (H2S), a metabolic byproduct of SRB, was considered the main causative agent that disrupted the morphology and function of gut epithelial cells. Associated study also showed that flagellin from Desulfovibrio vulgaris (DVF), the representative bacterium of the Desulfovibrio genus, could exacerbate colitis due to the interaction of DVF and LRRC19, leading to the secretion of pro-inflammatory cytokines. However, we still have limited understanding about the change of gut microbiota (GM) composition caused by overgrowth of SRB and its exacerbating effects on colitis. RESULTS: In this study, we transplanted D. vulgaris into the mice treated with or without DSS, and set a one-week recovery period to investigate the impact of D. vulgaris on the mice model. The outcomes showed that transplanted D. vulgaris into the normal mice could cause the gut inflammation, disrupt gut barrier and reduce the level of short-chain fatty acids (SCFAs). Moreover, D. vulgaris also significantly augmented DSS-induced colitis by exacerbating the damage of gut barrier and the secretion of inflammatory cytokines, for instance, IL-1ß, iNOS, and TNF-α. Furthermore, results also showed that D. vulgaris could markedly change GM composition, especially decrease the relative abundance of SCFAs-producing bacteria. Additionally, D. vulgaris significantly stimulated the growth of Akkermansia muciniphila probably via its metabolic byproduct, H2S, in vivo. CONCLUSIONS: Collectively, this study indicated that transplantation of D. vulgaris could cause gut inflammation and aggravate the colitis induced by DSS.

How Often Should Microbial Contamination Be Detected in Aircraft Fuel Systems? An Experimental Test of Aluminum Alloy Corrosion Induced by Sulfate-Reducing Bacteria.

Microbial contamination in aircraft fuel-containing systems poses significant threats to flight safety and operational integrity as a result of microbiologically influenced corrosion (MIC). Regular monitoring for microbial contamination in these fuel systems is essential for mitigating MIC risks. However, the frequency of monitoring remains a challenge due to the complex environmental conditions encountered in fuel systems. To investigate the impact of environmental variables such as water content, oxygen levels, and temperature on the MIC of aluminum alloy in aircraft fuel systems, orthogonal experiments with various combinations of these variables were conducted in the presence of sulfate-reducing bacteria. Among these variables, water content in the fuel oil demonstrated the most substantial influence on the corrosion rate of aluminum alloys, surpassing the effects of oxygen and temperature. Notably, the corrosion rate of aluminum alloys was the highest in an environment characterized by a 1:1 water/oil ratio, 0% oxygen, and a temperature of 35 °C. Within this challenging environment, conducive to accelerated corrosion, changes in the corrosion behavior of aluminum alloys over time were analyzed to identify the time point at which MIC intensified. Observations revealed a marked increase in the depth and width of corrosion pits, as well as in the corrosion weight-loss rate, starting from the 7th day. These findings offer valuable insights for determining the optimal frequency of microbial contamination detection in aircraft fuel systems.

Bacteremia caused by Desulfovibrio desulfuricans with the intestinal tract as the portal of entry: two case reports and a literature review.

BACKGROUND: Desulfovibrio desulfuricans (D. desulfuricans), a commensal anaerobic gram-negative rod endemic to the soil environment and human gastrointestinal tract, rarely causes bloodstream infections. We report two rare cases of bacteremia caused by D. desulfuricans in which the intestinal tract was the portal of entry. In addition, we summarize findings on D. desulfuricans. CASE PRESENTATION: Case 1: A 51-year-old man presented to the emergency department with the chief complaints of fever and right lower abdominal pain. He was admitted to the hospital with ascending colonic diverticulitis and received empirical antibacterial therapy with piperacillin/tazobactam. Blood culture revealed D. desulfuricans. The patient was discharged after 2 weeks of antimicrobial therapy. Case 2: A 95-year-old woman presented to our hospital with a chief complaint of fever. Owing to an elevated inflammatory response and pyuria, the patient was diagnosed with pyelonephritis and treated with ceftriaxone. Klebsiella pneumoniae was detected in her urine culture, while D. desulfuricans was detected in her blood culture. The patient was then treated with ampicillin/sulbactam for 14 days. The fecal occult blood test result was positive, suggesting a colonic mucosal lesion, such as a malignant tumor, may have been the portal of entry for D. desulfuricans bacteremia. Previous literature reviews indicate that D. desulfuricans bacteremia often results from liver or renal abscesses, intestinal lesions, among others, serving as the portal of entry. Although no specific underlying disease has been reported, it is more common in the older population. We encountered two cases of D. desulfuricans bacteremia and combined them with 15 cases from previous studies to explore the characteristics of the disease. The proportion of patients aged [Formula: see text]60 years was 73.7%; overall, 73.7% had gastrointestinal complications, and 63.2% had abdominal symptoms at the time of presentation. CONCLUSIONS: We encountered two rare cases of D. desulfurican bacteremia. This type of bacteremia is more common in elderly people over 60 years of age and is often associated with hepatobiliary and gastrointestinal diseases.

Sulfidation of Cd-Sch during the microbial sulfate reduction: Nanoscale redistribution of Cd.

Schwertmannite (Sch) is found in environments abundant in iron and sulfate. Microorganisms that utilize iron or sulfate can induce the phase transition of Schwertmannite, consequently leading to the redistribution of coexisting pollutants. However, the impact of the molar ratio of sulfate to iron (S/Fe) on the microbial-mediated transformation of Schwertmannite and its implications for the fate of cadmium (Cd) have not been elucidated. In this study, we examined how S/Fe influenced mineral transformation and the fate of Cd during microbial reduction of Cd-loaded Schwertmannite by Desulfovibrio vulgaris. Our findings revealed that an increase in the S/Fe ratio facilitated sulfate-reducing bacteria (SRB) in mitigating the toxicity of Cd, thereby expediting the generation of sulfide (S(-II)) and subsequently triggering mineral phase transformation. As the S/Fe ratio increased, the predominant minerals in the system transitioned from prismatic-cluster vivianite to rose-shaped mackinawite. The Cd phase and distribution underwent corresponding alterations. Cd primarily existed in its oxidizable state, with its distribution being directly linked not only to FeS content but also showing a robust correlation with phosphorus. The coexistence of vivianite and FeS minerals proved to be more favorable for Cd immobilization. These findings have significant implications for understanding the biogeochemistry of iron (oxyhydr)oxides and Cd fate in anaerobic environments.

Restricted intake of sulfur-containing amino acids reversed the hepatic injury induced by excess Desulfovibrio through gut-liver axis.

Diet is a key player in gut-liver axis. However, the effect of different dietary patterns on gut microbiota and liver functions remains unclear. Here, we used rodent standard chow and purified diet to mimic two common human dietary patterns: grain and plant-based diet and refined-food-based diet, respectively and explored their impacts on gut microbiota and liver. Gut microbiota experienced a great shift with notable increase in Desulfovibrio, gut bile acid (BA) levels elevated significantly, and liver inflammation was observed in mice fed with the purified diet. Liver inflammation and elevated gut BA levels also occurred in mice fed with the chow diet after receiving Desulfovibrio desulfuricans ATCC 29,577 (DSV). Restriction of sulfur-containing amino acids (SAAs) prevented liver injury mainly through higher hepatic antioxidant and detoxifying ability and reversed the elevated BA levels due to excess Desulfovibrio. Ex vivo fermentation of human fecal microbiota with primary BAs demonstrated that DSV enhanced production of secondary BAs. Higher concentration of both primary and secondary BAs were found in the gut of germ-free mice after receiving DSV. In conclusion, Restriction of SAAs in diet may become an effective dietary intervention to prevent liver injury associated with excess Desulfovibrio in the gut.

Structural and biochemical characterization of the M405S variant of Desulfovibrio vulgaris formate dehydrogenase.

Molybdenum- or tungsten-dependent formate dehydrogenases have emerged as significant catalysts for the chemical reduction of CO2 to formate, with biotechnological applications envisaged in climate-change mitigation. The role of Met405 in the active site of Desulfovibrio vulgaris formate dehydrogenase AB (DvFdhAB) has remained elusive. However, its proximity to the metal site and the conformational change that it undergoes between the resting and active forms suggests a functional role. In this work, the M405S variant was engineered, which allowed the active-site geometry in the absence of methionine Sδ interactions with the metal site to be revealed and the role of Met405 in catalysis to be probed. This variant displayed reduced activity in both formate oxidation and CO2 reduction, together with an increased sensitivity to oxygen inactivation.

Effects of intestinal Desulfovibrio bacteria on host health and its potential regulatory strategies: A review.

Increasing studies have focused on the relationship between Desulfovibrio bacteria (DSV) and host health in recent years. However, little is known about the mechanisms by which DSV affects host health and the strategies to accurately regulate DSV numbers. This review mainly presents the relationship between DSV and host health, potential modulatory strategies, and the potential mechanisms affecting host health. Evidence suggests that DSV can both promote host health and induce the occurrence and development of disease, and these effects are closely related to its metabolites (e.g., H2S and short-chain fatty acids) and biofilm. DSV abundance in the intestine is influenced by probiotics, prebiotics, diet, lifestyle, and drugs.

Desulfovibrio desulfuricans and its derived metabolites confer resistance to FOLFOX through METTL3.

BACKGROUND: Chemoresistance is a critical factor contributing to poor prognosis in clinical patients with cancer undergoing postoperative adjuvant chemotherapy. The role of gut microbiota in mediating resistance to tumour chemotherapy remains to be investigated. METHODS: Patients with CRC were categorised into clinical benefit responders (CBR) and no clinical benefit responders (NCB) based on chemotherapy efficacy. Differential bacterial analysis using 16S rRNA sequencing revealed Desulfovibrio as a distinct microbe between the two groups. Employing a syngeneic transplantation model, we assessed the effect of Desulfovibrio on chemotherapy by measuring tumour burden, weight, and Ki-67 expression. We further explored the mechanisms underlying the compromised chemotherapeutic efficacy of Desulfovibrio using metabolomics, western blotting, colony formation, and cell apoptosis assays. FINDINGS: In comparison, Desulfovibrio was more abundant in the NCB group. In vivo experiments revealed that Desulfovibrio colonisation in the gut weakened the efficacy of FOLFOX. Treatment with Desulfovibrio desulfuricans elevates serum S-adenosylmethionine (SAM) levels. Interestingly, SAM reduced the sensitivity of CRC cells to FOLFOX, thereby promoting the growth of CRC tumours. These experiments suggest that SAM promotes the growth and metastasis of CRC by driving the expression of methyltransferase-like 3 (METTL3). INTERPRETATION: A high abundance of Desulfovibrio in the intestines indicates poor therapeutic outcomes for postoperative neoadjuvant FOLFOX chemotherapy in CRC. Desulfovibrio drives the manifestation of METTL3 in CRC, promoting resistance to FOLFOX chemotherapy by increasing the concentration of SAM. FUNDING: This study is supported by Wuxi City Social Development Science and Technology Demonstration Project (N20201005).

Combining metabolic flux analysis with proteomics to shed light on the metabolic flexibility: the case of Desulfovibrio vulgaris Hildenborough.

Introduction: Desulfovibrio vulgaris Hildenborough is a gram-negative anaerobic bacterium belonging to the sulfate-reducing bacteria that exhibits highly versatile metabolism. By switching from one energy mode to another depending on nutrients availability in the environments" it plays a central role in shaping ecosystems. Despite intensive efforts to study D. vulgaris energy metabolism at the genomic, biochemical and ecological level, bioenergetics in this microorganism remain far from being fully understood. Alternatively, metabolic modeling is a powerful tool to understand bioenergetics. However, all the current models for D. vulgaris appeared to be not easily adaptable to various environmental conditions. Methods: To lift off these limitations, here we constructed a novel transparent and robust metabolic model to explain D. vulgaris bioenergetics by combining whole-cell proteomic analysis with modeling approaches (Flux Balance Analysis). Results: The iDvu71 model showed over 0.95 correlation with experimental data. Further simulations allowed a detailed description of D. vulgaris metabolism in various conditions of growth. Altogether, the simulations run in this study highlighted the sulfate-to-lactate consumption ratio as a pivotal factor in D. vulgaris energy metabolism. Discussion: In particular, the impact on the hydrogen/formate balance and biomass synthesis is discussed. Overall, this study provides a novel insight into D. vulgaris metabolic flexibility.

Biomineralization of Sulfate-Reducing Bacteria In Situ-Induced Preparation of Nano Fe2O3-Fe(Ni)S/C as High-Efficiency Oxygen Evolution Electrocatalyst.

Transition metal-based catalysts possess high catalytic activity for oxygen evolution reaction (OER). However, the preparation of high-performance OER electrocatalysts using simple strategies with a low cost still faces a major challenge. Herein, this work presents an innovative, in situ-induced preparation of the Fe2O3, FeS, and NiS nanoparticles, supported on carbon blacks (CBs) (denoted as Fe2O3-Fe(Ni)S/C) as a high-efficiency oxygen evolution electrocatalyst by employing biomineralization. Biomineralization, a simple synthesis strategy, demonstrates a huge advantage in controlling the size of the Fe2O3 and Fe(Ni)S nanoparticles, as well as achieving uniform nanoparticle distribution on carbon blacks. It is found that the electrocatalyst Fe2O3-Fe(Ni)S/C-200 shows a good OER electrocatalytic activity with a small loading capacity, and it has a small overpotential and Tafel slope in 1 m KOH solution with values of 264 mV and 42 mV dec-1, respectively, at a current density of 10 mA cm-2. Additionally, it presents good electrochemical stability for over 24 h. The remarkable and robust electrocatalytic performance of Fe2O3-Fe(Ni)S/C-200 is attributed to the synergistic effect of Fe2O3, FeS, and doped-Ni species as well as its distinct 3D spherical structure. This approach indicates the promising applications of biomineralization for the bio-preparation of functional materials and energy conversion.

Moringa oleifera leaf alleviates functional constipation via regulating the gut microbiota and the enteric nervous system in mice.

Moringa oleifera Lam. leaf is not only a new food resource in China, but also a traditional medicinal plant. It is commonly used in the folk to alleviate constipation, but its laxative mechanism is not fully understood. Hence we investigated it in loperamide-induced functional constipation (FC) mice. The results showed that MOAE significantly regulated not only gastrointestinal hormones and neurotransmitters in serum but also important gastrointestinal motility factors in the enteric nervous system (ENS)-interstitial cells of Cajal (ICCs)-smooth muscle cell (SMC) network. Meanwhile, MOAE attenuated intestinal inflammation, increased cecal short-chain fatty acid levels and colonic antimicrobial peptide expression, and improved the impaired intestinal barrier function in loperamide-induced FC mice. In addition, MOAE also increased fecal water content by inhibiting the mRNA expression of colonic aquaporins (Aqp3 and Aqp4) in FC mice. Interestingly and importantly, MOAE affected the intestinal microbiota by inhibiting some key "constipation-causing" microbiota, such as Bacteroidaceae, Clostridiaceae, Bacteroides, and Ruminococcus, and promoting the growth of other important "constipation-curing" microbiota, such as Butyricoccus, Tyzzerella, and Desulfovibrio. These important taxa are significantly associated with a variety of indicators of constipation. These findings suggest that MOAE can promote defecation through its rich chemical composition to modulate the ENS-ICCs-SMCs network and the gut microecosystem.

Descripción de un caso de sepsis abdominal por Desulfovibrio desulfuricans / Description of a case of abdominal sepsis due to Desulfovibrio desulfuricans

Resumen Desulfovibrio spp. son bacterias anaerobias estrictas, ubicuas en la naturaleza, quepueden formar parte del tracto gastrointestinal humano o animal, pero también son bacteriasambientales presentes en el suelo y en el agua. Pueden persistir de manera asintomática enel intestino o comportarse como patógenos oportunistas, asociados con bacteriemia primariae infecciones intraabdominales. El número de infecciones por Desulfovibrio spp. puede estarsubestimado debido a su lenta velocidad de crecimiento y a que muchos laboratorios no realizancultivos en anaerobiosis de manera rutinaria. Pruebas sencillas, como el examen de la movilidaden fresco y de la morfología celular en la coloración de Gram, sumadas a la presencia de SH2en agar SIM y a la observación de una fluorescencia roja a pH alcalino bajo luz UV, seríanindicativas de Desulfovibrio spp. Se describe el caso de una bacteriemia por Desulfovibriodesulfuricans en una mujer con cuadro clínico de sepsis abdominal por apendicitis gangrenosacon fallo multiorgánico.

Abstract Desulfovibrio spp. are strict anaerobes that are ubiquitous in nature. They can reside in the human or animal gastrointestinal tract and, as they are also environmental bacteria, may be present in soil and water. They can persist asymptomatically in the intestine or behave as opportunistic pathogens associated with primary bacteremia and intraabdominal infections. Several Desulfovibrio spp. infections may be underestimated due to their slow growth rate and because many laboratories do not routinely perform anaerobic cultures. Simple tests such as motility detection on a fresh subculture, Gram stain to confirm cell morphology, presence of H2S in SIM agar and production of a red fluorescence in alkaline pH under UV light would be indicative of Desulfovibrio spp.

Effects of cadmium sulfide nanoparticles on sulfate bioreduction and oxidative stress in Desulfovibrio desulfuricans.

Sulfate-containing wastewater has a serious threat to the environment and human health. Microbial technology has great potential for the treatment of sulfate-containing wastewater. It was found that nano-photocatalysts could be used as extracellular electron donors to promote the growth and metabolic activity of non-photosynthetic microorganisms. However, nano-photocatalysts could also induce oxidative stress and damage cells. Therefore, the interaction mechanism between photosynthetic nanocatalysts and non-photosynthetic microorganisms is crucial to determine the regulatory strategies for microbial wastewater treatment technologies. In this paper, the mechanism and regulation strategy of cadmium sulfide nanoparticles (CdS NPs) on the growth of sulfate-reducing bacteria and the sulfate reduction process were investigated. The results showed that the sulfate reduction efficiency could be increased by 6.4% through CdS NPs under light conditions. However, the growth of Desulfovibrio desulfuricans C09 was seriously inhibited by 55% due to the oxidative stress induced by CdS NPs on cells. The biomass and sulfate reduction efficiency could be enhanced by 6.8% and 5.9%, respectively, through external addition of humic acid (HA). At the same time, the mechanism of the CdS NPs strengthening the sulfate reduction process by sulfate bacteria was also studied which can provide important theoretical guidance and technical support for the development of microbial technology combined with extracellular electron transfer (EET) for the treatment of sulfate-containing wastewater.

Desulfovibrio vulgaris as a model microbe for the study of corrosion under sulfate-reducing conditions.

Corrosion of iron-containing metals under sulfate-reducing conditions is an economically important problem. Microbial strains now known as Desulfovibrio vulgaris served as the model microbes in many of the foundational studies that developed existing models for the corrosion of iron-containing metals under sulfate-reducing conditions. Proposed mechanisms for corrosion by D. vulgaris include: (1) H2 consumption to accelerate the oxidation of Fe0 coupled to the reduction of protons to H2; (2) production of sulfide that combines with ferrous iron to form iron sulfide coatings that promote H2 production; (3) moribund cells release hydrogenases that catalyze Fe0 oxidation with the production of H2; (4) direct electron transfer from Fe0 to cells; and (5) flavins serving as an electron shuttle for electron transfer between Fe0 and cells. The demonstrated possibility of conducting transcriptomic and proteomic analysis of cells growing on metal surfaces suggests that similar studies on D. vulgaris corrosion biofilms can aid in identifying proteins that play an important role in corrosion. Tools for making targeted gene deletions in D. vulgaris are available for functional genetic studies. These approaches, coupled with instrumentation for the detection of low concentrations of H2, and proven techniques for evaluating putative electron shuttle function, are expected to make it possible to determine which of the proposed mechanisms for D. vulgaris corrosion are most important.

Desulfovibrio vulgaris, a potent acetic acid-producing bacterium, increased by Astragalus polysaccharides to attenuate nonalcoholic fatty liver disease in mice / 中国药理学与毒理学杂志

OBJECTIVE Although the underlying mechanism is largely unknown, gut dysbiosis has emerged as a central initiator of obesity-related diseases including nonalcoholic fatty liver disease (NAFLD), type 2 diabetes and meta?bolic syndrome. The emerging evidence support the use of prebiotics like herb-derived polysaccharides for treating NAFLD by modulating gut microbiome. So, our study focused on the microbiota-dependent anti-NAFLD effect and the exact mechanisms of Astragalus polysaccharides (APS) extracted from Astragalus mongholicus Bunge in high-fat diet (HFD) fed mice. METHODS Co-housing experiment was used to assess the microbiota dependent anti-NAFLD effect of APS. Then, targeted metabolomics and metagenomics were adopted for determining short-chain fatty acids (SCFAs) and bacteria that were specifically enriched by APS. Further in vitro experiment was carried out to test the capacity of SCFAs-producing of identified bacterium. Finally, the anti-NAFLD efficacy of identified bacterium was tested in HFD fed mice. RESULTS Our results first demonstrated the anti-NAFLD effect of APS in HFD fed mice and the contribution of gut microbiota. Moreover, our results indicated that SCFAs, predominantly acetic acid were elevated in APS-supplemented mice and ex vivo experiment. Metagenomics revealed that D. vulgaris from Desulfovibrio genus was not only enriched by APS, but also a potent generator of acetic acid, which showed significant anti-NAFLD effects in HFD fed mice. In addition, D. vulgaris modulated the hepatic gene expression pattern of lipids metabolism, particularly suppressed hepatic fatty acid synthase (FASN) and CD36 protein expression. CONCLUSION APS enriched D. vulgaris is effective on attenuating hepatic steatosis possibly through producing acetic acid, and modulation on hepatic lipids metabolism in mice. Further studies are warranted to explore the long-term impacts of D. vulgaris on host metabolism and the underly?ing mechanism.

Análise de limas endodônticas submetidas a biocorrosão por bactérias redutoras de sulfato in vitro / Analysis of endodontic files submitted to biocorrosion by sulfate reducing bacteria in vitro

Objetivo: Avaliar as alterações químicas presentes na superfície metálica de limas endodônticas fraturadas em canais radiculares, in vitro, após a inoculação intrarradicular de culturas de BRS de três cepas microbianas, Desulfovibrio desulfuricans (uma cepa oral e outra ambiental) e Desulfovibrio fairfieldensis. Métodos: foram analisadas 5 limas kerr #90, sendo uma Lima Kerr nova, sem tratamento, e as outras 4 limas fraturadas dentro de canais radiculares in vitro, com posterior inoculação de Desulfovibrio desulfuricans, cepa oral e ambiental, e Desulfovibrio fairfieldensis e um grupo controle sem inoculação bacteriana, por 477 dias. Os grupos foram analisados no modo EDS (Espectrometria de Energia Dispersiva de Raios-x) do microscópio eletrônico de varredura (FEI-Inspect-S50). Resultados:A presença do S, Cl e O foram relacionados ao processo biocorrosivo, assim como a redução dos elementos de liga nesta área. Conclusão:As análises no modo EDS demonstraram biocorrosão ao longo da superfície metálica das limas quando empregado o biofármaco BACCOR, nas três diferentes cepas empregadas, indicada pela redução dos elementos formadores da liga metálica, Fe, Ni e Cr, com a associação da presença de elementos indicadores de biocorrosão como O, Cl e S. (AU)

Aim:To evaluate the chemical alterations present on the metallic surface of root canal fractured endodontic files in vitro after the intraradicular inoculation of BRS cultures of three microbial strains, Desulfovibrio desulfuricans (one oral and one environmental strain), and Desulfovibrio fairfieldensis. Methods: Five kerr #90 files were analyzed, one new untreated Kerr file and the other 4 files fractured within root canals in vitro, with a subsequent inoculation of Desulfovibrio desulfuricans (oral and environmental strains), and Desulfovibrio fairfieldensis, as well as a control group without bacterial inoculation for 477 days. The groups were analyzed using the scanning electron microscope (FEI-Inspect-S50) EDS (X-ray Dispersive Energy Spectrometry) mode. Results:The presence of S, Cl, and O were related to the biocorrosive process, as well as the reduction of alloying elements in this area. Conclusion: The EDS mode analysis showed biocorrosion along the metallic surface of the files when the BACCOR biopharmaceutical was used in the three different strains employed in this study, indicated by the reduction of the alloying elements ­ Fe, Ni, and Cr ­ with the association of the presence of indicator elements of biocorrosion, such as O, Cl, and S. (AU)

Anaerobiospirillum succiniciproducens y Desulfovibrio desulfuricans en 2 casos de bacteriemias insidiosas / Anaerobiospirillum succiniciproducens and Desulfovibrio desulfuricans in 2 cases of insidious bacteremia

Se presentan 2 casos de bacteriemias insidiosas por bacilos gram negativos anaerobios curvos, espiralados, móviles e infrecuentes en pacientes atendidos en un hospital de la ciudad de Buenos Aires. Estas bacteriemias, asociadas al aislamiento de Anaerobiospirillum y Desulfovibrio, fueron de origen poco claro y afectaron a pacientes inmunocomprometidos, con patologías simultáneas. Pruebas claves en la identificación del género Anaerobiospirillum fueron el estudio de la micromorfología, su carácter de anaerobio estricto, el resultado negativo en la prueba de catalasa, el patrón de discos de interés taxonómico, la fermentación de glucosa y la producción de β-N-acetilglucosaminidasa. El género Desulfovibrio se diferenció por el perfil presentado en las pruebas con discos, por ser asacarolítico, sin actividad de enzimas glucosídicas, y por producir desulfoviridina y H2S. Se alerta sobre la resistencia o sensibilidad intermedia de Anaerobiospirillum succiniciproducens (especie a la que correspondió el aislado de Anaerobiospirillum) a algunos de los antimicrobianos de primera línea frente a bacilos gram negativos anaerobios, como el metronidazol; fueron activas las combinaciones de aminopenicilinas con inhibidores de β-lactamasas y el imipenem. Desulfovibrio desulfuricans (especie a la que correspondió el aislado de Desulfovibrio) fue productora de β-lactamasas y resistente a las cefalosporinas; en cambio, fueron activos el metronidazol, el imipenem y la levofloxacina. La identificación confiable de estos microorganismos orienta hacia el mejor esquema terapéutico.

Two cases of insidious bacteremia by uncommon curve and spiral-shaped, motile anaerobic gram-negative rods are presented. Both of them were of an unclear origin and occurred in immunosuppressed patients with simultaneous diseases. The key tests for the identification of Anaerobiospirillum were its micromorphology, a strictly anaerobic condition, negative catalase activity, the special-potency disk profile, glucose fermentation, and β-NAG production. Desulfovibrio species was identified by all the above preliminary tests but with a different disk profile, as well as for being asaccharolytic and desulfoviridin and H2S producer. We here alert about the resistance or intermediate susceptibility of Anaerobiospirillum succiniciproducens against antimicrobial agents, such as metronidazole, one of the first-line drugs used for the treatment of anaerobic gram-negative infections. Aminopenicillins with β-lactamase-inhibitor combinations and imipenem were active for this agent. Desulfovibrio desulfuricans was β-lactamase producer and resistant to cephalosporins, while metronidazole, imipenem and levofloxacin were active. A reliable identification of these microorganisms is important for establishing the best therapeutic scheme.