scholarly journals Continuous cultivation of the lithoautotrophic nitrate-reducing Fe(II)-oxidizing culture KS in a chemostat bioreactor

2021 ◽  
Author(s):  
Timm Bayer ◽  
Elizabeth Tomaszewski ◽  
Casey Bryce ◽  
Andreas Kappler ◽  
James Byrne

Laboratory-based studies on microbial Fe(II) oxidation are commonly performed over just a few weeks in small volumes with high substrate concentrations, resulting in geochemical gradients and volumetric effects caused by sampling. We used a chemostat to enable uninterrupted supply of medium, and investigated autotrophic growth of the nitrate-reducing Fe(II)-oxidizing culture KS for 24 days. We analysed Fe- and N-speciation, cell-mineral associations, and the identity of minerals. Results were compared to different batch systems (50 and 700 ml – static/shaken). The Fe(II) oxidation rate was highest in the chemostat with 7.57 mM Fe(II) d-1, while the extent was similar (averaged 92% of all Fe(II)). Short-range ordered Fe(III) phases, presumably ferrihydrite, precipitated and later goethite was detected in the chemostat. 1 mM solid phase Fe(II) remained in the chemostat, up to 15 µM of reactive nitrite was measured, and 42% of visualized cells were partially or completely mineral-encrusted, likely caused by abiotic oxidation of Fe(II) by nitrite. Despite (partial) encrustation, cells were still viable. Our results show that even with similar oxidation rates as in batch cultures, cultivating Fe(II)-oxidizing microorganisms under continuous conditions reveals mechanistic insights on the role of reactive intermediates for Fe(II) oxidation, mineral formation and cell-mineral interactions.

1997 ◽  
Vol 36 (10) ◽  
pp. 27-36 ◽  
Author(s):  
P. Mungkarndee ◽  
S. M. Rao Bhamidimarri ◽  
A. J. Mawson ◽  
R. Chong

Biodegradation of the mixed inhibitory substrates, 2,4-dichlorophenoxyacetic acid (2,4-D) and para-chloro-ortho-cresol (PCOC) was studied in aerobic batch cultures. Each substrate added beyond certain concentrations inhibited the degradation of the other. This mutual inhibition was found to be enhanced by 2,4-dichlorophenol (2,4-DCP) which is an intermediate metabolic product of 2,4-D. When 2,4-DCP accumulated to approximatelY 40 mg/l degradation of all compounds in the mixed 2,4-D and PCOC substrate system was completely inhibited. The degradation of 2,4-D and PCOC individually was also found to be inhibited by elevated concentrations of 2,4-DCP added externally, while PCOC inhibited the utilization of the intermediate.


Author(s):  
David L. Kirchman

Geomicrobiology, the marriage of geology and microbiology, is about the impact of microbes on Earth materials in terrestrial systems and sediments. Many geomicrobiological processes occur over long timescales. Even the slow growth and low activity of microbes, however, have big effects when added up over millennia. After reviewing the basics of bacteria–surface interactions, the chapter moves on to discussing biomineralization, which is the microbially mediated formation of solid minerals from soluble ions. The role of microbes can vary from merely providing passive surfaces for mineral formation, to active control of the entire precipitation process. The formation of carbonate-containing minerals by coccolithophorids and other marine organisms is especially important because of the role of these minerals in the carbon cycle. Iron minerals can be formed by chemolithoautotrophic bacteria, which gain a small amount of energy from iron oxidation. Similarly, manganese-rich minerals are formed during manganese oxidation, although how this reaction benefits microbes is unclear. These minerals and others give geologists and geomicrobiologists clues about early life on Earth. In addition to forming minerals, microbes help to dissolve them, a process called weathering. Microbes contribute to weathering and mineral dissolution through several mechanisms: production of protons (acidity) or hydroxides that dissolve minerals; production of ligands that chelate metals in minerals thereby breaking up the solid phase; and direct reduction of mineral-bound metals to more soluble forms. The chapter ends with some comments about the role of microbes in degrading oil and other fossil fuels.


Coatings ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 503
Author(s):  
Jaehyun Lee ◽  
Ehsan Esmaili ◽  
Giho Kang ◽  
Baekhoon Seong ◽  
Hosung Kang ◽  
...  

The dimple occurs by sudden pressure inversion at the droplet’s bottom interface when a droplet collides with the same liquid-phase or different solid-phase. The air film entrapped inside the dimple is a critical factor affecting the sequential dynamics after coalescence and causing defects like the pinhole. Meanwhile, in the coalescence dynamics of an electrified droplet, the droplet’s bottom interfaces change to a conical shape, and droplet contact the substrate directly without dimple formation. In this work, the mechanism for the dimple’s suppression (interfacial change to conical shape) was studied investigating the effect of electric pressure. The electric stress acting on a droplet interface shows the nonlinear electric pressure adding to the uniform droplet pressure. This electric stress locally deforms the droplet’s bottom interface to a conical shape and consequentially enables it to overcome the air pressure beneath the droplet. The electric pressure, calculated from numerical tracking for interface and electrostatic simulation, was at least 108 times bigger than the air pressure at the center of the coalescence. This work helps toward understanding the effect of electric stress on droplet coalescence and in the optimization of conditions in solution-based techniques like printing and coating.


2011 ◽  
Vol 14 (2) ◽  
pp. 12-17 ◽  
Author(s):  
Tatiana Vladimirovna Saprina ◽  
F E Lazarenko ◽  
T S Prokhorenko ◽  
N V Ryazantseva ◽  
Irina Nikolaevna Vorozhtsova

Aim. To elucidate the role of Th1/Th2 polarization of immune response in LADA patients in the realization of the clinical phenotype of the disease. Materials and methods. 70 patients aged 21-61 (mean 41.3?1.0 yr) with DM diagnosed based on WHO criteria (1999). Groups 1 and 2 included 13 DM1and 57 DM2 patients (34.6?7.2 and 43.6?7.6 yr respectively). 27 DM2 patients (41.2?1.6 yr) presumably had LADA (P. Zimmet's criteria).Serum anti-GAD65, ICA, and IAA antibodies along with C-peptide were measured in fasting sera and 120 min after GTT by solid phase immunoenzymeassays following manufacturer's instructions with the use of a photometer for Multiscan EX microplates (ThermoLabSystems, Finland) at405 nm (for GAG and ICA) and 450 nm (for IAA and C-peptide). GAD, IAA, and C-peptides levels were calculated automatically from calibrationcurves. Mononuclear leukocytes were isolated by centrifugation in the ficoll-verographin density gradient. The cells thus obtained were resuspendedin the complete nutritient medium reducing their concentration to 2.0x10^6/ml. Phytohemagglutinin (Difco, Germany) was added (10 mcg/1 ml) tothe samples to stimulate mononuclear leukocytes; cell suspensions were further incubated for 24 hr. Initial and PGA-induced levels of IL-2, 4, 10 insupernatants of cell cultures were measured by solid phase immunoassay at 450 nm. Results. At least one type of autoantibodies (GAD, ICA or IAA) was identified in 24.3% of all DM patients (17/70) and in 18% of the DM2 patients(10/57). The level of anti-GAD and ICA ABs and percentage of AB-positive patients were higher in the LADA group while that of anti-IAA ABs amongDM1 patients without LADA. Two AB types at a time were found in 17% (4/23) of the patients with autoimmune DM in the absence of significantdifference between LADA and DM1. Patients with LADA had a significantly lower basal C-peptide level than DM2 patients. The was a tendencytoward lower level of stimulated C-peptide secretion in LADA patients compared with DM2 ones. It suggests impairment of beta-cell secretory functionaffected by the autoimmune process. We observed enhanced basal production of IFN-y by blood mononuclear leukocytes in all DM patients in theabsence of significant difference between the groups. Mitogen-activated production in all CD patients was lower than normal without inter-groupdifferences. Patients with DM2 had the inverted type of IL-2 secretion unlike those with autoimmune diabetes. In both cases it was significantly differentfrom normal values. There was a tendency toward higher basal production of IL-4 by mononuclear leukocytes in LADA and DM2 comparedwith CD1 which reflects pathogenetic peculiarities of beta-cell function in LADA differing from those in DM1 and responsible for slower impairment ofbeta-cell function in this condition. Basal and PGA- induced production of IL-10 was higher in LADA and DM2 than in DM1. It suggests enhancedsuppressor activity of leukocytes that may protect beta-cells from autoimmune destruction and determines gradual development of clinical symptoms ofinsulin deficiency. In contrast, low production of IL-10 in DM1 gives evidence of polarization of the immune response. Conclusion. The loss of functional parenchyma and manifestation of insulin deficiency in LADA occur at a relatively low rate due to the peculiarcharacter of cytokine-mediated cell interactions. It suggests the necessity of an active and careful diagnostic strategy with the use of immunologicalmethods for examination of elder patients presenting with a variety of pathogenetic variants of DM.


2020 ◽  
Vol 11 (4) ◽  
pp. 5225-5228
Author(s):  
Deepshikha Verma ◽  
Pillai V N R ◽  
Giriraj Tailor

Protecting groups like Fmoc and coupling both steps are essential to monitoring the Fmoc SPPS (Solid Phase Peptide Synthesis) reaction completion. Reliable methods are used to detect the unreacted number of amino groups for monitoring these two essential reaction steps of coupling and cleavage. The ability to detect the complete coupling, incomplete coupling or failure of coupling we use many colour tests in the laboratory and based on this the Fmoc peptide chemistry allows the control of the completion of the Fmoc cleavage. The most important test used is the Kaiser test and highly recommended to monitor the coupling and cleavage steps. If the result of colour tests is positive after coupling, then the second coupling should be performed. Then again use the colour test to detect the level of coupling. If the result is still slightly positive, repeat coupling with the smaller modification of reagents such as used PyBOP instead of HOBT AND HOAT. These colour tests help in revealing the presence of unreacted amino-functional groups. Thus, we need to block these free N-terminal of amino- acids which help in avoiding the making of deletion of sequence.


2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Claudia Curci ◽  
Fabio Sallustio ◽  
Nada Chaoul ◽  
Angela Picerno ◽  
Gabriella Lauriero ◽  
...  

Abstract Background and Aims The IgA nephropathy (IgAN) is the most frequent primitive glomerulonephritis. In the last years, the role of mucosal immunity in IgAN, together with that of the gut microbiota in the activation of innate and adaptive immune cells, has gained importance. Particularly interesting is the role of the microbiota and intestinal immunity in IgAN. BAFF and APRIL can be produced by the intestinal epithelium, in response to signals triggered by TLRs once activated by the commensal bacteria present in the intestinal lumen, representing the link between microbiota and intestinal immunity. To date, even if hypothesized, this relationship in IgAN patients has not been investigated. Here, we studied the intestinal-renal axis connections analyzing levels of BAFF, April and intestinal-activated B cells in IgAN patients. Method Serum and fecal samples were collected from 44 IgAN patients, 22 non-IgA glomerulonephritides (controls) and 22 healthy subjects (HS) with similar clinical features. BAFF and APRIL serum levels were measured by ELISA assay. Metabolomic analysis of fecal microbiome was performed using Biochrom 30 series amino acid analyzer and gas-chromatography mass spectrometry/solid-phase microextraction (GC-MS/SPME) analysis. B cell subsets were investigated by FACS. Results IgAN patients had increased serum levels of BAFF cytokine compared to the control group of patients with non-IgA glomerulonephritis and compared with HS (p<0.0001and p=0.012, respectively). We found that serum BAFF levels positively correlated with the levels of 24h-proteinuria in IgAN patients (r2 = 0.2269, p <0.001). We correlated serum BAFF levels with fecal concentration of 5 different metabolites of 30 IgAN patients, which were previously investigated for the fecal microbiota. These organic compounds had been found at significantly higher levels in the feces of IgAN patients compared to HS. Serum BAFF levels positively correlated with the levels of fecal metabolites: 4-(1,1,3,3-tetramethylbutyl) phenol (r2 = 0.2882, p = 0.0027), p-tert-butyl-phenol (r2 = 0.386, p = 0.0003), methyl neopentyl phthalic acid (r2 = 0.3491, p =0.0007), hexadecyl ester benzoic acid (r2 = 0.2832, p =0.003) and furanone A (r2 = 0.1743, p = 0.024). Serum levels of APRIL were significantly increased in IgAN patients respect to control groups (4.49 ± 0.54 vs 2.27 ± 1 ng/ml, p=0.0014). We found a correlation between APRIL and serum creatinine (r2 = 0.159, p =0.04) and eGFR (r2 = 0.2395, p =0.0082), while no correlation was found between APRIL and fecal metabolite levels in IgAN patients. In addition, we found that subjects with IgAN have a significantly higher proportion of circulating Bregs, Memory B cells and IgA secreting-plasmablasts activated at the intestinal level (CCR9+INTB7+) compared to HS. Conclusion The results of our study showed for the first time an important correlation of serum levels of BAFF with intestinal microbiota in patients with IgAN, confirming the hypothesis of the pathogenic role of intestinal mucosal hyperresponsiveness in the IgAN patients. The intestinal-renal axis plays a crucial role in Berger's glomerulonephritis, whose complex pathogenesis may contribute several factors as genetics, pathogens and food.


2015 ◽  
Vol 75-76 ◽  
pp. 309-314
Author(s):  
M.E. Palumbo ◽  
R.G. Urso ◽  
Z. Kaňuchová ◽  
C. Scirè ◽  
M. Accolla ◽  
...  

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