Anaerobic Lactate Production Is Associated With Decreased Microcirculatory Blood Flow and Decreased Mitochondrial Respiration Following Cardiovascular Surgery With Cardiopulmonary Bypass.

OBJECTIVES: Quantify the relationship between perioperative anaerobic lactate production, microcirculatory blood flow, and mitochondrial respiration in patients after cardiovascular surgery with cardiopulmonary bypass. DESIGN: Serial measurements of lactate-pyruvate ratio (LPR), microcirculatory blood flow, plasma tricarboxylic acid cycle cycle intermediates, and mitochondrial respiration were compared between patients with a normal peak lactate (≤ 2 mmol/L) and a high peak lactate (≥ 4 mmol/L) in the first 6 hours after surgery. Regression analysis was performed to quantify the relationship between clinically relevant hemodynamic variables, lactate, LPR, and microcirculatory blood flow. SETTING: This was a single-center, prospective observational study conducted in an academic cardiovascular ICU. PATIENTS: One hundred thirty-two patients undergoing elective cardiovascular surgery with cardiopulmonary bypass. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients with a high postoperative lactate were found to have a higher LPR compared with patients with a normal postoperative lactate (14.4 ± 2.5 vs. 11.7 ± 3.4; p = 0.005). Linear regression analysis found a significant, negative relationship between LPR and microcirculatory flow index ( r = -0.225; ß = -0.037; p = 0.001 and proportion of perfused vessels: r = -0.17; ß = -0.468; p = 0.009). There was not a significant relationship between absolute plasma lactate and microcirculation variables. Last, mitochondrial complex I and complex II oxidative phosphorylation were reduced in patients with high postoperative lactate levels compared with patients with normal lactate (22.6 ± 6.2 vs. 14.5 ± 7.4 pmol O 2 /s/10 6 cells; p = 0.002). CONCLUSIONS: Increased anaerobic lactate production, estimated by LPR, has a negative relationship with microcirculatory blood flow after cardiovascular surgery. This relationship does not persist when measuring lactate alone. In addition, decreased mitochondrial respiration is associated with increased lactate after cardiovascular surgery. These findings suggest that high lactate levels after cardiovascular surgery, even in the setting of normal hemodynamics, are not simply a type B phenomenon as previously suggested.

Supercritical Fluid Extraction of Fucoxanthin from the Diatom Phaeodactylum tricornutum and Biogas Production through Anaerobic Digestion.

Phaeodactylum tricornutum is the marine diatom best known for high-value compounds that are useful in aquaculture and food area. In this study, fucoxanthin was first extracted from the diatom using supercritical fluid extraction (SFE) and then using the extracted diatom-like substrate to produce bioenergy through anaerobic digestion (AD) processes. Factors such as temperature (30 °C and 50 °C), pressure (20, 30, and 40 MPa), and ethanol (co-solvent concentration from 10% to 50% v/v) were optimized for improving the yield, purity, and recovery of fucoxanthin extracted using SFE. The highest yield (24.41% w/w) was obtained at 30 MPa, 30 °C, and 30% ethanol but the highest fucoxanthin purity and recovery (85.03mg/g extract and 66.60% w/w, respectively) were obtained at 30 MPa, 30 °C, and 40%ethanol. Furthermore, ethanol as a factor had the most significant effect on the overall process of SFE. Subsequently, P.tricornutum biomass and SFE-extracted diatom were used as substrates for biogas production through AD. The effect of fucoxanthin was studied on the yield of AD, which resulted in 77.15 ± 3.85 LSTP CH4/kg volatile solids (VS) and 56.66 ± 1.90 LSTP CH4/kg VS for the whole diatom and the extracted P.tricornutum, respectively. Therefore, P.tricornutuman can be considered a potential source of fucoxanthin and methane and both productions will contribute to the sustainability of the algae-biorefinery processes.

Complex networks analysis reinforces centrality hematological role on aerobic-anaerobic performances of the Brazilian Paralympic endurance team after altitude training.

This study investigated the 30-days altitude training (2500 m, LHTH-live and training high) on hematological responses and aerobic-anaerobic performances parameters of high-level Paralympic athletes. Aerobic capacity was assessed by 3000 m run, and anaerobic variables (velocity, force and mechanical power) by a maximal 30-s semi-tethered running test (AO30). These assessments were carried out at low altitude before (PRE) and after LHTH (5-6 and 15-16 days, POST1 and POST2, respectively). During LHTH, hematological analyzes were performed on days 1, 12, 20 and 30. After LHTH, aerobic performance decreased 1.7% in POST1, but showed an amazing increase in POST2 (15.4 s reduction in the 3000 m test, 2.8%). Regarding anaerobic parameters, athletes showed a reduction in velocity, force and power in POST1, but velocity and power returned to their initial conditions in POST2. In addition, all participants had higher hemoglobin (Hb) values at the end of LHTH (30 days), but at POST2 these results were close to those of PRE. The centrality metrics obtained by complex networks (pondered degree, pagerank and betweenness) in the PRE and POST2 scenarios highlighted hemoglobin, hematocrit (Hct) and minimum force, velocity and power, suggesting these variables on the way to increasing endurance performance. The Jaccard's distance metrics showed dissimilarity between the PRE and POST2 graphs, and Hb and Hct as more prominent nodes for all centrality metrics. These results indicate that adaptive process from LHTH was highlighted by the complex networks, which can help understanding the better aerobic performance at low altitude after 16 days in Paralympic athletes.

Energetics contribution during no-gi Brazilian jiu jitsu sparring and its association with regional body composition.

We used measurements of metabolic perturbation obtained after sparring to estimate energetics contribution during no-gi Brazilian jiu-jitsu. Ten advanced grapplers performed two six-minute sparring bouts separated by 24 hours. Kinetics of recovery rate of oxygen uptake was modelled and post-combat-sparring blood-lactate concentration measured to estimate oxygen equivalents for phospholytic and glycolytic components of anaerobic energetics, respectively. Linear regression was used to estimate end-combat-sparring rate of oxygen uptake. Regional and whole-body composition were assessed using dual X-ray absorptiometry with associations between these measurements and energy turnover explored using Pearson's correlation coefficient (significance, P < 0.05). Estimated oxygen equivalents for phospholytic and glycolytic contributions to anaerobic metabolism were 16.9 ± 8.4 (~28%) and 44.6 ± 13.5 (~72%) mL∙kg-1, respectively. Estimated end-exercise rate of oxygen uptake was 44.2 ± 7.0 mL∙kg-1∙min-1. Trunk lean mass was positively correlated with both total anaerobic and glycolytic-specific energetics (total, R = 0.645, p = 0.044; glycolytic, R = 0.692, p = 0.027) and negatively correlated with end-exercise rate of oxygen uptake (R = -0.650, p = 0.042). There were no correlations for any measurement of body composition and phospholytic-specific energetics. Six minutes of no-gi Brazilian jiu-jitsu sparring involves high relative contribution from the glycolytic component to total anaerobic energy provision and the link between this energetics profile and trunk lean mass is consistent with the predominance of ground-based combat that is unique for this combat sport. Training programs for Brazilian jiu-jitsu practitioners should be designed with consideration given to these specific energetics characteristics.

Conspicuous Smooth and White Egg-Shaped Sulfur Structures on a Deep-Sea Hydrothermal Vent Formed by Sulfide-Oxidizing Bacteria.

Conspicuous egg-shaped, white, and smooth structures were observed at a hydrothermal vent site in the Guaymas Basin, Gulf of California. The gelatinous structures decomposed within hours after sampling. Scanning electron microscopy (SEM) and light microscopy showed that the structure consisted of filaments of less than 0.1 µm thickness, similar to those observed for "Candidatus Arcobacter sulfidicus." SEM-energy-dispersive X-ray spectroscopy (EDS) showed that the filaments were sulfur rich. According to 16S rRNA gene amplicon and fluorescence in situ hybridization (FISH) analyses, Arcobacter, a sulfide oxidizer that is known to produce filamentous elemental sulfur, was among the dominant species in the structure and was likely responsible for its formation. Arcobacter normally produces woolly snowflake like structures in opposed gradients of sulfide and oxygen. In the laboratory, we observed sulfide consumption in the anoxic zone of the structure, suggesting an anaerobic conversion. The sulfide oxidation and decomposition of the structure in the laboratory may be explained by dissolution of the sulfur filaments by reaction with sulfide under formation of polysulfides. IMPORTANCE At the deep-sea Guaymas Basin hydrothermal vent system, sulfide-rich hydrothermal fluids mix with oxygenated seawater, thereby providing a habitat for microbial sulfur oxidation. Microbial sulfur oxidation in the deep sea involves a variety of organisms and processes and can result in the excretion of elemental sulfur. Here, we report on conspicuous white and smooth gelatinous structures found on hot vents. These strange egg-shaped structures were often observed on previous occasions in the Guaymas Basin, but their composition and formation process were unknown. Our data suggest that the notable and highly ephemeral structure was likely formed by the well-known sulfide-oxidizing Arcobacter. While normally Arcobacter produces loose flocs or woolly layers, here smooth gel-like structures were found.

Unveiling the role of EVs in anaerobic parasitic protozoa.

The anaerobic or microaerophilic protozoan parasites such as the enteric human pathogens Entamoeba histolytica, Giardia intestinalis, Cryptosporidium parvum, Blastocystis hominis and urogenital tract parasites Trichomonas vaginalis are able to survival in an environment with oxygen deprivation. Despite living in hostile environments these pathogens adopted different strategies to survive within the hosts. Among them, the release of extracellular vesicles (EVs) has become an active endeavor in the study of pathogenesis for these parasites. EVs are heterogenous, membrane-limited structures that have played important roles in cellular communication, transferring information through cargo and modulating the immune system of the host. In this review, we described several aspects of the recently characterized EVs of the anaerobic protozoa, including their role in adhesion, modulation of the immune response and omics analysis to understand the potential of these EVs in the pathogenesis of these diseases caused by anaerobic parasites.

Phages in Anaerobic Systems.

Since the discovery of phages in 1915, these viruses have been studied mostly in aerobic systems, or without considering the availability of oxygen as a variable that may affect the interaction between the virus and its host. However, with such great abundance of anaerobic environments on the planet, the effect that a lack of oxygen can have on the phage-bacteria relationship is an important consideration. There are few studies on obligate anaerobes that investigate the role of anoxia in causing infection. In the case of facultative anaerobes, it is a well-known fact that their shifting from an aerobic environment to an anaerobic one involves metabolic changes in the bacteria. As the phage infection process depends on the metabolic state of the host bacteria, these changes are also expected to affect the phage infection cycle. This review summarizes the available information on phages active on facultative and obligate anaerobes and discusses how anaerobiosis can be an important parameter in phage infection, especially among facultative anaerobes.

Effects of Beta-Alanine Supplementation on Physical Performance in Aerobic-Anaerobic Transition Zones: A Systematic Review and Meta-Analysis.

Beta-alanine supplementation (BA) has a positive impact on physical performance. However, evidence showing a benefit of this amino acid in aerobic-anaerobic transition zones is scarce and the results controversial. The aim of this systematic review and meta-analysis is to analyze the effects of BA supplementation on physical performance in aerobic-anaerobic transition zones. At the same time, the effect of different dosages and durations of BA supplementation were identified. The search was designed in accordance with the PRISMA® guidelines for systematic reviews and meta-analyses and performed in Web of Science (WOS), Scopus, SPORTDiscus, PubMed, and MEDLINE between 2010 and 2020. The methodological quality and risk of bias were evaluated with the Cochrane Collaboration tool. The main variables were the Time Trial Test (TTT) and Time to Exhaustion (TTE) tests, the latter separated into the Limited Time Test (LTT) and Limited Distance Test (LDT). The analysis was carried out with a pooled standardized mean difference (SMD) through Hedges' g test (95% CI). Nineteen studies were included in the systematic review and meta-analysis, revealing a small effect for time in the TTT (SMD, -0.36; 95% CI, -0.87-0.16; I2 = 59%; p = 0.010), a small effect for LTT (SMD, 0.25; 95% CI, -0.01-0.51; I2 = 0%; p = 0.53), and a large effect for LDT (SMD, 4.27; 95% CI, -0.25-8.79; I2 = 94%; p = 0.00001). BA supplementation showed small effects on physical performance in aerobic-anaerobic transition zones. Evidence on acute supplementation is scarce (one study); therefore, exploration of acute supplementation with different dosages and formats on physical performance in aerobic-anaerobic transition zones is needed.

Occurrence of Aspergillus niger strains on a polychrome cotton painting and their elimination by anoxic treatment.

This study aimed to isolate and identify the population of filamentous fungi colonizing a cotton painting, whose conservation status was compromised and showed signs of biodeterioration due to dirt accumulation and microbial metabolism. In addition, microbiological techniques such as cultivation-dependent approach and molecular biology were used to identify microbial populations and to eliminate their metabolic action. For this, the nondestructive anoxic atmosphere technique was used, in which the microbial metabolism was affected by the absence of oxygen. Prior to exposure to an anoxic atmosphere, only one fungal species, Aspergillus niger, was identified at 12 points sampled in the obverse and reverse of the artwork; no fungal species persisted as a result of anoxic treatment. These results showed that exposure to anoxic conditions was effective for the total elimination of isolated fungal strains as well as their spores. In conclusion, this study proved the unprecedented effectiveness of a nondestructive technique for artwork on textile colonized by black fungi species. Thus, this interdisciplinary work involving conservation, microbiology, and chemistry presents a tool to eliminate microorganisms, while maintaining the integrity of artwork and safety of the restorer, that can be applied prior to artwork restoration.

Cycling Performance Enhancement After Drop Jumps May Be Attributed to Postactivation Potentiation and Increased Anaerobic Capacity.

de Poli, RAB, Boullosa, DA, Malta, ES, Behm, D, Lopes, VHF, Barbieri, FA, and Zagatto, AM. Cycling performance enhancement after drop jumps may be attributed to postactivation potentiation and increased anaerobic capacity. J Strength Cond Res 34(9): 2465-2475, 2020-The study aimed to investigate the effects of drop jumps (DJs) on supramaximal cycling performance, anaerobic capacity (AC), electromyography, and fatigue. Thirty-eight recreational cyclists participated into 3 independent studies. In study 1 (n = 14), neuromuscular fatigue was assessed with the twitch interpolation technique. In study 2 (n = 16), the AC and metabolic contributions were measured with the maximal accumulated oxygen deficit method and the sum of the glycolytic and phosphagen pathways. In study 3 (n = 8), postactivation potentiation (PAP) induced by repeated DJs was evaluated. The DJ protocol was effective for significantly improving cycling performance by +9.8 and +7.4% in studies 1 and 2, respectively (p ≤ 0.05). No differences were observed in electromyography between conditions (p = 0.70); however, the force evoked by a doublet at low (10 Hz) and high frequencies (100 Hz) declined for control (-16.4 and -23.9%) and DJ protocols (-18.6 and -26.9%) (p < 0.01). Force decline was greater in the DJ condition (p < 0.03). Anaerobic capacity and glycolytic pathway contributions were +7.7 and +9.1% higher after DJ protocol (p = 0.01). Peak force during maximal voluntary contraction (+5.6%) and doublet evoked force at 100 Hz (+5.0%) were higher after DJs. The DJ protocol induced PAP, improved supramaximal cycling performance, and increased AC despite higher peripheral fatigue.

Anaerobic Degradation of Non-Methane Alkanes by "Candidatus Methanoliparia" in Hydrocarbon Seeps of the Gulf of Mexico.

Crude oil and gases in the seabed provide an important energy source for subsurface microorganisms. We investigated the role of archaea in the anaerobic degradation of non-methane alkanes in deep-sea oil seeps from the Gulf of Mexico. We identified microscopically the ethane and short-chain alkane oxidizers "Candidatus Argoarchaeum" and "Candidatus Syntrophoarchaeum" forming consortia with bacteria. Moreover, we found that the sediments contain large numbers of cells from the archaeal clade "Candidatus Methanoliparia," which was previously proposed to perform methanogenic alkane degradation. "Ca. Methanoliparia" occurred abundantly as single cells attached to oil droplets in sediments without apparent bacterial or archaeal partners. Metagenome-assembled genomes of "Ca. Methanoliparia" encode a complete methanogenesis pathway including a canonical methyl-coenzyme M reductase (MCR) but also a highly divergent MCR related to those of alkane-degrading archaea and pathways for the oxidation of long-chain alkyl units. Its metabolic genomic potential and its global detection in hydrocarbon reservoirs suggest that "Ca. Methanoliparia" is an important methanogenic alkane degrader in subsurface environments, producing methane by alkane disproportionation as a single organism.IMPORTANCE Oil-rich sediments from the Gulf of Mexico were found to contain diverse alkane-degrading groups of archaea. The symbiotic, consortium-forming "Candidatus Argoarchaeum" and "Candidatus Syntrophoarchaeum" are likely responsible for the degradation of ethane and short-chain alkanes, with the help of sulfate-reducing bacteria. "Ca. Methanoliparia" occurs as single cells associated with oil droplets. These archaea encode two phylogenetically different methyl-coenzyme M reductases that may allow this organism to thrive as a methanogen on a substrate of long-chain alkanes. Based on a library survey, we show that "Ca. Methanoliparia" is frequently detected in oil reservoirs and may be a key agent in the transformation of long-chain alkanes to methane. Our findings provide evidence for the important and diverse roles of archaea in alkane-rich marine habitats and support the notion of a significant functional versatility of the methyl coenzyme M reductase.

Differences between genders in anaerobic capacity during a supramaximal effort

Aim: The present study aimed to verify if there is a difference between genders in anaerobic capacity estimated by energetic equivalents of glycolytic and phosphagen pathways (AC[La-]+EPOCfast). Methods: In this way, 8 men and 8 women (physical education students) were subjected to the following sequence of tests: session 1) graded exercise test to measure the maximal oxygen consumption (VÖ½ O2max) and intensity associated with VÖ½ O2max (iVÖ½ O2max); sessions 2 to 3) familiarization with supramaximal effort at 115% of iVÖ½ O2max; session 4) supramaximal effort at 115% of iVÖ½ O2max to measure AC[La-]+EPOCfast. Results: The AC[La-]+EPOCfast was lower in women compared to men when expressed in absolute and relative values (-38.11%; p=0.01 and -25.71%; p=0.03, respectively). A non-significant difference was observed in performance in the supramaximal effort (-12.08%; p=0.15), besides which, a likely negative inference was observed when comparing women to men. In addition, energetic equivalents of the glycolytic pathway (e[La- ]) were also lower in women when expressed in relative and absolute values (-47.01%; p=0.001 and -36.71%; p=0.001, respectively), however no statistical difference was found for energetic equivalents of the phosphagen pathway (ePCr) (p>0.05). Conclusion: The AC[La-]+EPOCfast is lower in women compared to men, mainly due to differences in the glycolytic pathway.(AU)

Anaerobic capacity estimated by the sum of both oxygen equivalents from the glycolytic and phosphagen pathways is dependent on exercise mode: Running versus cycling.

The purpose of this study was to verify whether the exercise modality (i.e., running and cycling) alters the magnitude of "anaerobic" capacity estimated by a single supramaximal effort (AC[La]+EPOCfast). Fourteen healthy men (age: 26±9 years) underwent a maximum incremental test and a supramaximal effort to exhaustion at 115% of the intensity associated with maximal oxygen uptake to determine the AC[La]+EPOCfast (i.e., the sum of both oxygen equivalents from the glycolytic and phosphagen pathways), performed on both a treadmill and cycle ergometer. The maximal oxygen uptake during running was higher (p = 0.001; large effect size) vs. cycling (49.2±3.8 mL·kg-1·min-1 vs. 44.7±5.7 mL·kg-1·min-1, respectively). Contrarily, the oxygen equivalent from the glycolytic metabolism was not different between exercise modalities (p = 0.133; small effect size; running = 2.27±0.51 L and cycling = 2.33±0.49 L). Furthermore, the "anaerobic" capacity was likely meaningfully (3.9±0.6 L and 54.1±6.0 mL·kg-1) and very likely meaningfully greater in running than cycling (3.6±0.7 L and 49.2±6.1 mL·kg). Additionally, the contribution of the phosphagen metabolism was higher (p = 0.001; large effect size) for running compared to cycling (1.6±0.3 L vs.1.3±0.3 L respectively). Therefore, the "anaerobic" capacity estimated by the sum of both oxygen equivalents from the glycolytic and phosphagen pathways during a supramaximal effort is influenced by exercise modality and is able to identify the difference in phosphagen metabolic contribution, based on the methodological conditions of this study.

The Ammonium Channel NOD26 is the Evolutionary Innovation that Drives the Emergence, Consolidation, and Dissemination of Nitrogen-Fixing Symbiosis in Angiosperms.

Increasing evidence indicates that N-fixing symbiosis has evolved several times in the N-fixing clade of angiosperms and that this evolution is driven by a single evolutionary innovation. However, the genetics of this ancestral predisposition to N-fixing symbiosis remains unclear. A natural candidate for such molecular innovation is the ammonium channel NOD26, the main protein component of the symbiosome membrane, which facilitates the plant uptake of the nitrogen fixed by symbiotic bacteria. Here, in concordance with the emergence of N-fixing symbiosis in angiosperms but not in ancestral plants, phylogenetic analysis showed that NOD26 belongs to an angiosperm-exclusive subgroup of aquaporins. Integrated genomic, phylogenetic, and gene expression analyses supported NOD26 occurrence in the N-fixing clade, the increase in the NOD26 copy number by block and tandem duplications in legumes, and the low-copy number or even the loss of NOD26 in non-legume species of the N-fixing clade, which correlated with the possibility to lose N-fixing symbiosis in legume and non-legume lineages. Metabolic reconstructions showed that retention of NOD26 in N-fixing precursor could represent an adaptive mechanism to bypass energy crisis during anaerobic stress by ammonium detoxification. Finally, we discuss the potential use of NOD26 to transfer N-fixation to non-N-fixing crops as cereals.

Paenibacillus helianthi sp. nov., a nitrogen fixing species isolated from the rhizosphere of Helianthus annuus L.

Three facultatively anaerobic endospore-forming bacteria were isolated from the rhizosphere of sunflowers grown in fields of Rio Grande do Sul State, Brazil. The designated type strain P26ET was previously identified as a sunflower growth promoting bacterium and is able to fix nitrogen and to excrete ammonia. According to analyses of 16S rRNA gene sequences, P26ET presented similarity values above 98.8% in relation to Paenibacillus azotifigens NF2-4-5T, Paenibacillus graminis RSA19T, Paenibacillus jilunlii Be17T, Paenibacillus salinicaeni LAM0A28T, and Paenibacillus sonchi X19-5T. Phylogenetic reconstructions based on 16S rRNA gene and core proteome data showed that the strains P26ET, P3E and P32E form a distinct clade, which did not include any type strain of the currently described Paenibacillus species. Also, genomic comparisons using average nucleotide identity (ANI), Orthologous ANI and in silico DNA-DNA hybridization revealed similarity ranges below the recommended thresholds when the three isolates from sunflower were compared to their close relatives. The DNA G + C content of strain P26ET was determined to be 49.4 mol%. The major cellular fatty acids are anteiso-C15:0 and iso-C15:0, representing about 58 and 14% of the total fatty acids in P26ET, respectively. Based on different taxonomic genomic metrics, phylogeny, and phenotypic data, we propose that strain P26ET (= DSM 102269 = BR10509) represents a novel species within the genus Paenibacillus, for which the name Paenibacillus helianthi sp. nov. is proposed.

Graphene oxide triggers mass transfer limitations on the methanogenic activity of an anaerobic consortium with a particulate substrate.

Graphene oxide (GO) is an emerging nanomaterial widely used in many manufacturing applications, which is frequently discharged in many industrial effluents eventually reaching biological wastewater treatment systems (WWTS). Anaerobic WWTS are promising technologies for renewable energy production through biogas generation; however, the effects of GO on anaerobic digestion are poorly understood. Thus, it is of paramount relevance to generate more knowledge on these issues to prevent that anaerobic WWTS lose their effectiveness for the removal of pollutants and for biogas production. The aim of this work was to assess the effects of GO on the methanogenic activity of an anaerobic consortium using a particulate biopolymer (starch) and a readily fermentable soluble substrate (glucose) as electron donors. The obtained results revealed that the methanogenic activity of the anaerobic consortium supplemented with starch decreased up to 23-fold in the presence of GO compared to the control incubated in the absence of GO. In contrast, we observed a modest improvement on methane production (>10% compared to the control lacking GO) using 5 mg of GO L-1 in glucose-amended incubations. The decrease in the methanogenic activity is mainly explained by wrapping of starch granules by GO, which caused mass transfer limitation during the incubation. It is suggested that wrapping is driven by electrostatic interactions between negatively charged oxygenated groups in GO and positively charged hydroxyl groups in starch. These results imply that GO could seriously hamper the removal of particulate organic matter, such as starch, as well as methane production in anaerobic WWTS.

Anaerobic Metabolism in T4 Acanthamoeba Genotype.

Members of the genus Acanthamoeba are of the most common protozoa that has been isolated from a variety of environment and affect immunocompromised individuals, causing granulomatous amoebic encephalitis and skin lesions. Acanthamoeba, in immunocompetent patients, may cause a keratitis related to corneal microtrauma. These free-living amoebas easily adapt to the host environment and wield metabolic pathways such as the energetic and respiratory ones in order to maintain viability for long periods. The energetic metabolism of cysts and trophozoites remains mostly unknown. There are a few reports on the energetic metabolism of these organisms as they are mitochondriate eukaryotes and some studies under aerobic conditions showing that Acanthamoeba hydrolyzes glucose into pyruvate via glycolysis. The aim of this study was to detect the energetic metabolic pathways with emphasis on anaerobic metabolism in trophozoites of three isolates of Acanthamoeba sp belonging to the T4 genotype. Two samples were collected in the environment and one was a clinical sample. The evaluation of these microorganisms proceeded as follows: rupture of trophozoites (7.5 × 103 parasites/ml) and biochemical analysis with high performance liquid chromatography and spectrophotometry. The anaerobic glycolysis was identified through the detection of glucose, pyruvate, and lactate. The protein catabolism was identified through the detection of fumarate, urea, and creatinine. The fatty acid oxidation was identified through the detection of acetate, beta-hydroxybutyrate, and propionate. The detected substances are the result of the consumption of energy reserves such as glycogen and lipids. The anaerobic glycolysis and protein catabolism pathways were observed in all three isolates: one clinical and two environmental. This study represents the first report of energetic pathways used by trophozoites from different isolates of the T4 genotype Acanthamoeba.

Characterization of a Marine Microbial Community Used for Enhanced Sulfate Reduction and Copper Precipitation in a Two-Step Process.

Marine microorganisms that are obtained from hydrothermal vent sediments present a great metabolic potential for applications in environmental biotechnology. However, the work done regarding their applications in engineered systems is still scarce. Hence, in this work, the sulfate reduction process carried out by a marine microbial community in an upflow anaerobic sludge blanket (UASB) reactor was investigated for 190 days under sequential batch mode. The effects of 1000 to 5500 mg L-1 of SO4-2 and the chemical oxygen demand (COD)/SO4-2 ratio were studied along with a kinetic characterization with lactate as the electron donor. Also, the feasibility of using the sulfide produced in the UASB for copper precipitation in a second column was studied under continuous mode. The system presented here is an alternative to sulfidogenesis, particularly when it is necessary to avoid toxicity to sulfide and competition with methanogens. The bioreactor performed better with relatively low concentrations of sulfate (up to 1100 mg L-1) and COD/SO4-2 ratios between 1.4 and 3.6. Under the continuous regime, the biogenic sulfide was sufficient to precipitate copper at a removal rate of 234 mg L-1 day-1. Finally, the identification of the microorganisms in the sludge was carried out; some genera of microorganisms identified were Desulfitobacterium and Clostridium.

Expanding the regulatory network that controls nitrogen fixation in Sinorhizobium meliloti: elucidating the role of the two-component system hFixL-FxkR.

In Sinorhizobium meliloti, nitrogen fixation is regulated in response to oxygen concentration through the FixL-FixJ two-component system (TCS). Besides this conserved TCS, the field isolate SM11 also encodes the hFixL-FxkR TCS, which is responsible for the microoxic response in Rhizobium etli. Through genetic and physiological assays, we evaluated the role of the hFixL-FxkR TCS in S. meliloti SM11. Our results revealed that this regulatory system activates the expression of a fixKf orthologue (fixKa), in response to low oxygen concentration. Null mutations in either hFixL or FxkR promote upregulation of fixK1, a direct target of FixJ. Furthermore, the absence of this TCS translates into higher nitrogen fixation values as well as higher expression of fixN1 in nodules. Individual mutations in each of the fixK-like regulators encoded in the S. meliloti SM11 genome do not completely restrict fixN1 or fixN2 expression, pointing towards redundancy among these regulators. Both copies of fixN are necessary to achieve optimal levels of nitrogen fixation. This work provides evidence that the hFixL-FxkR TCS is activated in response to low oxygen concentration in S. meliloti SM11 and that it negatively regulates the expression of fixK1, fixN1 and nitrogen fixation.