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All biological hydroxylation reactions are thought to derive the oxygen atom from one of three inorganic oxygen donors, O2, H2O2 or H2O. Here, we have identified the organic compound prephenate as the oxygen donor for the three hydroxylation steps of the O2-independent biosynthetic pathway of ubiquinone, a widely distributed lipid coenzyme. Prephenate is an intermediate in the aromatic amino acid pathway and genetic experiments showed that it is essential for ubiquinone biosynthesis in Escherichia coli under anaerobic conditions. Metabolic labeling experiments with 18O-shikimate, a precursor of prephenate, demonstrated the incorporation of 18O atoms into ubiquinone. The role of specific iron--sulfur enzymes belonging to the widespread U32 protein family is discussed. Prephenate-dependent hydroxylation reactions represent a unique biochemical strategy for adaptation to anaerobic environments. [ABSTRACT FROM AUTHOR]

Photogeneration of hydrogen in microalgae is thought to be among the mechanisms increasing their resilience to stresses including those caused by nutrient deprivation by re-routing the flow of electrons and reducing power in the cell. Metabolism of phosphorus (P), an essential nutrient, and its reserve forms such as polyphosphate (PolyP), is affected by and plays a role in the responses to diverse stresses, too. However, the potential interplay of the capability of photogeneration of hydrogen and turnover of phosphorus-rich inclusions in stressed microalgae cells so far escaped the attention of researchers. Here, we present a quantitative ultrastructural view of the turnover of P-rich inclusions in the model microalga Chlamydomonas reinhardtii strains, the parent strain CC-425 and its hydEF-1 mutant lacking hydrogenase activity as a function of sulfur and oxygen availability in the medium. In addition to the electron microscopy cell image analysis of the studied strains, we followed the elemental composition of the inclusions in different (sub) compartments of the cells obtained with energy dispersive X-ray spectroscopy. The stress caused by sulfur deprivation and subsequent transition of the microalgae culture to anaerobic conditions declined the size of phosphorus-containing inclusions but increased their number in the parent strain. Overall, the accumulation of the phosphorus-rich inclusions in hydEF-1 mutant was much lower than in the fully functional parent strain regardless of the cultivation conditions. We believe that impaired hydrogenase activity and correspondingly reduced sink of electrons and reducing power in the mutant strain indirectly affects the turnover of P and its reserves in the cell. These effects were manifested by the changes in the abundance, morphology, and elemental composition of the P-containing inclusions. We hypothesized that the sulfur-deprivation stress increased the initiation of the biosynthesis of PolyP chains, but their elongation and hence the formation of large PolyP-containing inclusions was hindered by anaerobiosis. [ABSTRACT FROM AUTHOR]

Molasses are a source of fermentable sugars, the substrate utilized for this study, as it is a subproduct from sugarcane processing for sucrose recovery. These sugars can be metabolized for bioH 2 production by bacterial strains, particularly strict-anaerobe Clostridium acetobutylicum (Clac) and its media preparation requires chemical of O 2 removal and N 2 flushing to promote anaerobiosis. Bacillus subtilis (Bsub) can deplete O 2 from culture media and promotes anaerobiosis, which is an aspect that would benefit from further research. Clac was grown in an anaerobic medium and produced 269 mL of hydrogen at 72 h. By co-culturing Clac and Bsub , 502 mL of bioH 2 were produced in a cysteine/N 2 flushed medium, while 203 mL of bioH 2 at 72 h were produced by aerobic co-culture. Additionally, in an untreated/aerobic conventional growth culture for Clac , no production of H 2 was detected, reinforcing the utility of Bsub for anaerobiosis generation. • Clostridium acetobutylicum consumes substrate approximately 6x faster than Bacillus subtilis. • Clostridium acetobutylicum produced 202 mL of bioH 2 in a culture with anaerobiosis assisted by Bacillus subtilis. • Bacillus subtilis CBB5555 can act as a biological O 2 depleter for anaerobic bioprocesses. [ABSTRACT FROM AUTHOR]

Anaeramoebae is a recently described phylum of anaerobic, marine amoebae, and amoeboflagellates belonging to the Metamonada supergroup. So far, six species have been described based on light microscopic morphology and sequences of the SSU rRNA gene. Here we present three new strains of Anaeramoeba with a description of their morphology, ultrastructure, and phylogenetic position based on the analysis of SSU rRNA gene sequences. Two of the strains represent a new species, Anaeramoeba pumila sp. nov., that has the smallest cells of all known Anaeramoeba species, and one that represents a species from the newly recognized Anaeramoeba flamelloides complex. Anaeramoebae are known to have a syntrophic relationship with prokaryotes. Our strains display two novel, remarkable types of symbioses, previously unknown from Anaeramoebae. [ABSTRACT FROM AUTHOR]

Green hydrogen, produced during microalgal photosynthesis, is regarded as one of the most promising sustainable energy sources. It utilizes sunlight and water, which are essentially unlimited, and its combustion results in only water as a waste product. In microalgal hydrogen energy production systems, the sensitivity of hydrogenase to O2 poses a significant challenge, limiting sustained photosynthetic H2 production in microalgae. Additionally, efficient photosynthetic H2 production in anaerobic microalgal cells is hindered by impaired electron source (photosystem II) and electron loss through the Calvin‐Benson cycle, cyclic electron transfer around photosystem I, and O2 photoreduction, which are identified as the other key challenges. Over the past eight decades, considerable progress has been made in addressing these challenges and regulating electron transfer to achieve sustainable and efficient photosynthetic H2 production in microalgae. In this review, we discuss a range of regulatory methods for achieving sustainable and efficient photosynthetic H2 production in microalgae. Emphasizing the significant progress made over the past eight decades, we also address current challenges and propose potential future solutions. [ABSTRACT FROM AUTHOR]

Phage therapy is a promising alternative in treating bacterial infections; however, understanding how bacteria and phages interact according to the environment is crucial. Although most phage therapy applications are intended for anaerobic environments (such as intestine or wounds), research on the effects of oxygen absence on the phage infection process is scarce . Here, we studied the effect of anoxic conditions in a Salmonella sp. – phage model, given the prevalence of this bacterial pathogen in anaerobic environments, and the potential of phage therapy for its control. Our results show that oxygen-deprived conditions affect the life cycle length of the phage and its capacity to control the bacterial population. Changes in host cell size associated with the FtsZ protein were observed, along with a modified emergence of resistance against the phage under different oxygen conditions. Interestingly, transcriptomic analyses revealed that phage infection in aerobic conditions triggers the expression of genes associated with nitrate reduction, an alternative pathway for anaerobic respiration. Meanwhile, uninfected anaerobic cultures displayed the upregulation of genes involved in a different anaerobic metabolic route comprising the transport of thiosulfate and sulfate. Altogether, our results provide insights into the importance of considering oxygen availability as an important factor for phage therapy when working with facultative anaerobic bacteria. IMPORTANCE Many parameters affect phage-bacteria interaction. Some of these parameters depend on the environment in which the bacteria are present. Anaerobiosis effect on phage infection in facultative anaerobic bacteria has not yet been studied. The absence of oxygen triggers metabolic changes in facultative bacteria and this affects phage infection and viral life cycle. Understanding how an anaerobic environment can alter the behavior of phages during infection is relevant for the phage therapy success. [ABSTRACT FROM AUTHOR]

Background: The endoplasmic reticulum (ER)-mitochondria membrane contact sites (MCS) are extensively studied in aerobic eukaryotes; however, little is known about MCS in anaerobes with reduced forms of mitochondria named hydrogenosomes. In several eukaryotic lineages, the direct physical tether between ER and the outer mitochondrial membrane is formed by ER-mitochondria encounter structure (ERMES). The complex consists of four core proteins (Mmm1, Mmm2, Mdm12, and Mdm10) which are involved in phospholipid trafficking. Here we investigated ERMES distribution in organisms bearing hydrogenosomes and employed Trichomonas vaginalis as a model to estimate ERMES cellular localization, structure, and function. Results: Homology searches revealed that Parabasalia-Anaeramoebae, anaerobic jakobids, and anaerobic fungi are lineages with hydrogenosomes that retain ERMES, while ERMES components were gradually lost in Fornicata, and are absent in Preaxostyla and Archamoebae. In T. vaginalis and other parabasalids, three ERMES components were found with the expansion of Mmm1. Immunofluorescence microscopy confirmed that Mmm1 localized in ER, while Mdm12 and Mmm2 were partially localized in hydrogenosomes. Pull-down assays and mass spectrometry of the ERMES components identified a parabasalid-specific Porin2 as a substitute for the Mdm10. ERMES modeling predicted a formation of a continuous hydrophobic tunnel of TvMmm1-TvMdm12-TvMmm2 that is anchored via Porin2 to the hydrogenosomal outer membrane. Phospholipid-ERMES docking and Mdm12-phospholipid dot-blot indicated that ERMES is involved in the transport of phosphatidylinositol phosphates. The absence of enzymes involved in hydrogenosomal phospholipid metabolism implies that ERMES is not involved in the exchange of substrates between ER and hydrogenosomes but in the unidirectional import of phospholipids into hydrogenosomal membranes. Conclusions: Our investigation demonstrated that ERMES mediates ER-hydrogenosome interactions in parabasalid T. vaginalis, while the complex was lost in several other lineages with hydrogenosomes. [ABSTRACT FROM AUTHOR]

The study of biodeterioration is an important issue to allow the best conservation and prevent the decay of cultural heritage and artworks. In Naples (Italy), a particular museum (Museodivino) preserves the miniature artworks representing Dante's Divine Comedy and Nativity scenes, executed with organic-based materials in walnut and clay shells. Since they showed putative signs of biodeterioration, the first aim of this study was to verify the presence of microbial colonization. A culture-dependent approach and molecular biology allowed us to isolate and identify the sole fungal strain Aspergillus NCCD (Nativity and Dante's Divine Comedy) belonging to the A. sydowii sub-clade. Based on this result, a sustainable and eco-friendly approach was applied to find a method to preserve the miniature artwork by contrasting the growth of the strain NCCD. Several essential oils used as a natural biocide were tested against Aspergillus strain NCCD belonging to the A. sydowii subclade to determine their potential antimicrobial activity. Results revealed that basil, cloves, fennel, and thyme essential oils exerted antifungal activity, although their effect depended also on the concentration used. Moreover, anoxic treatment and the control of the relative humidity were used in the presence of thyme, in vitro, and in vivo assays to define the impact on fungal growth. No fungal development was detected in vivo in the shells treated with thyme essential oil at high relative humidity after 60 days of incubation at 28 °C. These results highlighted that although relative humidity was the major factor affecting the development of the strain Aspergillus NDDC, the application of thyme in an anaerobic environment is essential in contrasting the fungal growth. Identifying the biodeterioration agent allowed us to plan an eco-friendly, non-destructive approach to be successfully used to guarantee the conditions suitable for conserving miniature artwork. [ABSTRACT FROM AUTHOR]

All-out exercise may not impair all central nervous system processes, such as those related to visual-motor abilities, and may actually prove stimulatory to such tasks allowing athletes and sports conditioning specialists to develop strategies to take advantage of/mitigate the effects of such exercise on athletic performance. Despite research indicating that visual-motor abilities play a critical role in athletic performance, research has primarily focused on the effect of all-out exercise on processes along the motor pathway, such as resultant force production or simple cognitive tasks. Such research has neglected to investigate the effect of all-out exercise on visual tasks. When investigations on visual tasks are forthcoming, they focus on prolonged aerobic exercise, which is not the primary metabolic pathway for all, or even the majority of sports. Sixty untrained males (experimental group; N = 30, control group; N = 30) completed a standardised six-item baseline sports vision test battery and one week later, the experimental participants returned to undertake a 30-second Wingate anaerobic test (30-WAnT) immediately followed by the same test battery. Significant (P < 0.05) improvements were found in accommodation facility, saccadic eye movement, speed of recognition, peripheral awareness and hand-eye coordination (P < 0.001 for all), but not visual memory (P = 0.242) following the 30-WAnT. Although the mechanisms underlying these improvements in visual task performance have not yet been studied, this study suggests that simple anaerobic all-out exercise does not cause central- or brain-based fatigue impairing the oculomotor system but may rather provide "excitability" of the underlying motor cortex, motoneurons and/or corticofugal connections utilised in visual task response. It appears that the sweeping improvements in visual task performance elucidate the need for an intense anaerobic warm-up when training visual skills and when visual skills form an integral part of athletic performance. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the nutritional changes in silages of four sorghum hybrids at different storage times. The design was a completely randomized in a 4 × 9 arrangement of plots with four replications. The four sorghum hybrids, Qualysilo, Chopper, Dominator, and Maxisilo, were allocated to the plots. In the subplots, sampling times were considered in fresh material and after 1, 3, 7, 14, 28, 56, 112, and 224 days of storage. The material was ensiled in experimental polyvinyl chloride silos and subsequently evaluated for chemical composition and fermentative profile over the nine different periods of the fermentation process. Crude protein was reduced by 1.21g/kg per day until the third day of fermentation. The levels of neutral detergent insoluble nitrogen and acid detergent insoluble nitrogen exceeded the upper limit of 311 g/kg of N. Of the studied silages, an NDF content of 734 g/kg was obtained in Maxisilo, which was higher than the other silages (average: 547 g/kg). Silage production can be achieved with the four materials; however, the Maxisilo sorghum provides silages of lower quality, with a DM content of less than 30%. The other materials fit into the nutritional profiles proposed by the literature, characterizing them as good quality silages. [ABSTRACT FROM AUTHOR]

Locomotion is a defining characteristic that can dictate many aspects of an organism's life history in the pursuit of maximizing fitness, including escaping predators, capturing prey, and transitioning between habitats. Exhaustive exercise can have negative consequences for both short-term and long-term energetics and life history trade-offs, influencing fish survival and reproduction. Studies of swimming performance and exhaustive exercise in fish are often conducted on individual species, but few multispecies analyses exist and even fewer in field settings. In fish, swimming performance and exercise have historically been studied in the laboratory using swim tunnels, but an increasing body of work in recreational fisheries science provides a novel way to examine swimming capacity and exhaustion. Using fight time, the time it takes for a hooked fish to be landed on rod and reel fishing gear, as an opportunistic proxy for fish exhaustion, a multispecies meta-analysis of data from studies on recreational fisheries was conducted to elucidate the factors that most influence capacity for exhaustive exercise. Data from 39 species of freshwater and marine fish were aggregated, and negative binomial mixed effects models as well as phylogenetic least squares regression were used to identify the factors that most influenced exhaustive exercise in the field. Fish total length, aspect ratio of the caudal fin, and body form were significant factors in explaining the capacity for exhaustive exercise. Large migratory fish with high aspect ratios were able to fight, and therefore exercise, the longest. These results illustrate that body form and physiology are both deeply intertwined to inform function across fish species and point to angling fight time as a useful approximation of fish swimming capabilities that can be further developed for understanding the limits of fish exercise physiology. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the effects of different proportions of cactus pear mucilage as a moistening additive on chemical composition, ammonia nitrogen, volatile fatty acids, microbial dynamics, fermentation losses, pH, and aerobic stability of corn grains in the form of silage throughout storage periods. The study was carried out in a completely randomized design with a factorial arrangement (5×2), with four replications. The factors corresponded to five forms of moistening for corn grain silage, 0, 5, 10, 20, and 40% of cactus pear mucilage (MA) associated or not with urea (U) in the level of 1.5% of DM. The lowest DM content was found in the silage moistened with 40% of cactus pear mucilage, and the greatest CP content was found in the same treatment but when urea was added. Lower contents of propionic and butyric acids were observed in the silages moistened with 5 and 10% of cactus pear without and with the addition of urea. Dry matter recovery was higher in the treatments with water and 10% of cactus pear. The use of urea affected the LAB population only at 15 d of silo opening, providing a bigger population (6.58 logs CFU g -1 ) when compared to silages without urea (6.27 logs CFU g -1 ). The lower concentrations of propionic and butyric acids observed in the silage moistened with 5 and 10% of cactus pear mucilage, regardless of the addition of urea. However, it is recommended to use 10% of cactus pear mucilage without the addition of urea, as it provides better indicators of good-quality silage. [ABSTRACT FROM AUTHOR]

BACKGROUND: The present study aimed to determine the optimal conditions for making wines by the carbonic maceration (CM) method. In this test, we attempted to identify the causes of the higher volatile acidity of some wines vinified by this method. Accordingly, we measured the development and speed of intracellular fermentation inside the whole grapes under different vinification conditions. RESULTS: An active fermentation of the must in the tanks produced by inoculation with active dry yeasts was more efficient for the process than the addition of exogenous carbon dioxide (CO2). In addition, in CM vinification, the moment of devatting had a great influence on the content of acetaldehyde and acetic acid in the whole grapes. CONCLUSION: Yeast inoculation in the tanks and control of the devatting time are technological tools that could play an important role in the characteristics of the wines produced by carbonic maceration, especially with respect to acetic acid content. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Vibrio cholerae can form biofilms in the aquatic environment and in the human intestine, facilitating the release of hyper-infectious aggregates. Due to the increasing antibiotic resistance, alternatives need to be found. One of these alternatives is antimicrobial peptides, including polymyxin B (PmB). In this study, we first investigated the resistance of V. cholerae O1 El Tor strain A1552 to various antimicrobials under aerobic and anaerobic conditions. An increased resistance to PmB is observed in anaerobiosis, with a 3-fold increase in the dose required for 50 % growth inhibition. We then studied the impact of the PmB on the formation and the degradation of V. cholerae biofilms to PmB. Our results show that PmB affects more efficiently biofilm formation under anaerobic conditions. On the other hand, preformed biofilms are susceptible to degradation by PmB at concentrations close to the minimal inhibitory concentration. At higher concentrations, we observe an opacification of the biofilm structures within 20 min post-treatment, suggesting a densification of the structure. This densification does not seem to result from the overexpression of matrix genes but rather from DNA release through massive cell lysis, likely forming a protective shield that limits the penetration of the PmB into the biofilm. [ABSTRACT FROM AUTHOR]

Mucor lusitanicus and some other members of the fungal order Mucorales display the phenomenon of morphological dimorphism. This means that these fungi aerobically produce filamentous hyphae, developing a coenocytic mycelium, but they grow in a multipolar yeast-like form under anaerobiosis. Revealing the molecular mechanism of the reversible yeast-hyphal transition can be interesting for both the biotechnological application and in the understanding of the pathomechanism of mucormycosis. In the present study, transcriptomic analyses were carried out after cultivating the fungus either aerobically or anaerobically revealing significant changes in gene expression under the two conditions. In total, 539 differentially expressed genes (FDR < 0.05, |log2FC| ≥ 3) were identified, including 190 upregulated and 349 downregulated transcripts. Within the metabolism-related genes, carbohydrate metabolism was proven to be especially affected. Anaerobiosis also affected the transcription of transporters: among the 14 up- and 42 downregulated transporters, several putative sugar transporters were detected. Moreover, a considerable number of transcripts related to amino acid transport and metabolism, lipid transport and metabolism, and energy production and conversion were proven to be downregulated when the culture had been transferred into an anaerobic atmosphere. [ABSTRACT FROM AUTHOR]

Background: Mitochondria and peroxisomes are the two organelles that are most affected during adaptation to microoxic or anoxic environments. Mitochondria are known to transform into anaerobic mitochondria, hydrogenosomes, mitosomes, and various transition stages in between, collectively called mitochondrion-related organelles (MROs), which vary in enzymatic capacity. Anaerobic peroxisomes were identified only recently, and their putatively most conserved function seems to be the metabolism of inositol. The group Archamoebae includes anaerobes bearing both anaerobic peroxisomes and MROs, specifically hydrogenosomes in free-living Mastigamoeba balamuthi and mitosomes in the human pathogen Entamoeba histolytica, while the organelles within the third lineage represented by Pelomyxa remain uncharacterized. Results: We generated high-quality genome and transcriptome drafts from Pelomyxa schiedti using single-cell omics. These data provided clear evidence for anaerobic derivates of mitochondria and peroxisomes in this species, and corresponding vesicles were tentatively identified in electron micrographs. In silico reconstructed MRO metabolism harbors respiratory complex II, electron-transferring flavoprotein, a partial TCA cycle running presumably in the reductive direction, pyruvate:ferredoxin oxidoreductase, [FeFe]-hydrogenases, a glycine cleavage system, a sulfate activation pathway, and an expanded set of NIF enzymes for iron-sulfur cluster assembly. When expressed in the heterologous system of yeast, some of these candidates localized into mitochondria, supporting their involvement in the MRO metabolism. The putative functions of P. schiedti peroxisomes could be pyridoxal 5′-phosphate biosynthesis, amino acid and carbohydrate metabolism, and hydrolase activities. Unexpectedly, out of 67 predicted peroxisomal enzymes, only four were also reported in M. balamuthi, namely peroxisomal processing peptidase, nudix hydrolase, inositol 2-dehydrogenase, and d-lactate dehydrogenase. Localizations in yeast corroborated peroxisomal functions of the latter two. Conclusions: This study revealed the presence and partially annotated the function of anaerobic derivates of mitochondria and peroxisomes in P. schiedti using single-cell genomics, localizations in yeast heterologous systems, and transmission electron microscopy. The MRO metabolism resembles that of M. balamuthi and most likely reflects the state in the common ancestor of Archamoebae. The peroxisomal metabolism is strikingly richer in P. schiedti. The presence of myo-inositol 2-dehydrogenase in the predicted peroxisomal proteome corroborates the situation in other Archamoebae, but future experimental evidence is needed to verify additional functions of this organelle. [ABSTRACT FROM AUTHOR]

Background: Mitochondria and peroxisomes are the two organelles that are most affected during adaptation to microoxic or anoxic environments. Mitochondria are known to transform into anaerobic mitochondria, hydrogenosomes, mitosomes, and various transition stages in between, collectively called mitochondrion-related organelles (MROs), which vary in enzymatic capacity. Anaerobic peroxisomes were identified only recently, and their putatively most conserved function seems to be the metabolism of inositol. The group Archamoebae includes anaerobes bearing both anaerobic peroxisomes and MROs, specifically hydrogenosomes in free-living Mastigamoeba balamuthi and mitosomes in the human pathogen Entamoeba histolytica, while the organelles within the third lineage represented by Pelomyxa remain uncharacterized. Results: We generated high-quality genome and transcriptome drafts from Pelomyxa schiedti using single-cell omics. These data provided clear evidence for anaerobic derivates of mitochondria and peroxisomes in this species, and corresponding vesicles were tentatively identified in electron micrographs. In silico reconstructed MRO metabolism harbors respiratory complex II, electron-transferring flavoprotein, a partial TCA cycle running presumably in the reductive direction, pyruvate:ferredoxin oxidoreductase, [FeFe]-hydrogenases, a glycine cleavage system, a sulfate activation pathway, and an expanded set of NIF enzymes for iron-sulfur cluster assembly. When expressed in the heterologous system of yeast, some of these candidates localized into mitochondria, supporting their involvement in the MRO metabolism. The putative functions of P. schiedti peroxisomes could be pyridoxal 5′-phosphate biosynthesis, amino acid and carbohydrate metabolism, and hydrolase activities. Unexpectedly, out of 67 predicted peroxisomal enzymes, only four were also reported in M. balamuthi, namely peroxisomal processing peptidase, nudix hydrolase, inositol 2-dehydrogenase, and d-lactate dehydrogenase. Localizations in yeast corroborated peroxisomal functions of the latter two. Conclusions: This study revealed the presence and partially annotated the function of anaerobic derivates of mitochondria and peroxisomes in P. schiedti using single-cell genomics, localizations in yeast heterologous systems, and transmission electron microscopy. The MRO metabolism resembles that of M. balamuthi and most likely reflects the state in the common ancestor of Archamoebae. The peroxisomal metabolism is strikingly richer in P. schiedti. The presence of myo-inositol 2-dehydrogenase in the predicted peroxisomal proteome corroborates the situation in other Archamoebae, but future experimental evidence is needed to verify additional functions of this organelle. [ABSTRACT FROM AUTHOR]

The waste generated by the mezcal and seafood restaurant industries has significant negative environmental impacts on soils, water, and air. However, these waste materials offer the potential for methane production through anaerobic digestion. This research focuses on evaluating a methodological strategy that involves using an enriched inoculum, co-digesting waste materials, and applying specific pretreatment (chemical, thermal, and adjusting the volumetric ratio of residues) to produce biogas with a high methane content, as assessed through the Biochemical Methane Potential (BMP) test. The results demonstrate that the enriched inoculum used in the test exhibited a pH of 8.18, an Alkalinity of 31 g CaCO3/L, and an alpha (α) ratio of 0.75. The BMP test was conducted over 29 days. In the most successful test, which involved pretreatment at a temperature of 50 °C, a residue ratio of 1 VSW (Volume of Seafood Waste)/2 VMV (Volume of Mezcal Vinasses), and a pH of 6.5, we achieved a methane production of 1420.51 ± 0.038 N mL CH4, with a biogas containing 81.5% methane. This process also yielded 175.37 N mL CH4 per gram of Volatile Solids (VS) and a degradation kinetic constant of 0.14 1/d. Through the methodological strategy used with the combination of an enriched and stabilized inoculum, the co-digestion of the waste for nutrient complementation, and the application of pretreatments, it was achieved that the different stages that constitute the anaerobic process allowed for the establishment of optimal conditions for the generation of biogas with a high methane content.Graphical Abstract: The waste generated by the mezcal and seafood restaurant industries has significant negative environmental impacts on soils, water, and air. However, these waste materials offer the potential for methane production through anaerobic digestion. This research focuses on evaluating a methodological strategy that involves using an enriched inoculum, co-digesting waste materials, and applying specific pretreatment (chemical, thermal, and adjusting the volumetric ratio of residues) to produce biogas with a high methane content, as assessed through the Biochemical Methane Potential (BMP) test. The results demonstrate that the enriched inoculum used in the test exhibited a pH of 8.18, an Alkalinity of 31 g CaCO3/L, and an alpha (α) ratio of 0.75. The BMP test was conducted over 29 days. In the most successful test, which involved pretreatment at a temperature of 50 °C, a residue ratio of 1 VSW (Volume of Seafood Waste)/2 VMV (Volume of Mezcal Vinasses), and a pH of 6.5, we achieved a methane production of 1420.51 ± 0.038 N mL CH4, with a biogas containing 81.5% methane. This process also yielded 175.37 N mL CH4 per gram of Volatile Solids (VS) and a degradation kinetic constant of 0.14 1/d. Through the methodological strategy used with the combination of an enriched and stabilized inoculum, the co-digestion of the waste for nutrient complementation, and the application of pretreatments, it was achieved that the different stages that constitute the anaerobic process allowed for the establishment of optimal conditions for the generation of biogas with a high methane content. [ABSTRACT FROM AUTHOR]

Introduction: This examination expects to decide the impact of the High-Intensity Interval Training technique utilizing Elevation Training Mask dependent on oxygen-consuming limit on the physiological capacities of futsal players. Methods: This research is an experimental study with a 2x2 factorial design. The conditional assumption test, followed by the Two way ANOVA test to propose a hypothesis or formulation. The problem in this study was with the help of the computerized SPSS application version 25. Twenty male futsal players (age 21.70 ± 0.80 years, height 165.75 ± 8.29 cm, weight 64.29 ± 4.98 kg, VO2max 45 ,3 ± 5.23 ml.kg-1.min-1). The period lasted six weeks, with each training group participating three days per week of equal volume and intensity. The post-test was conducted four days after the last training session. Results: No difference in the physiological abilities of futsal players based on the use of ETM and non-ETM. Male futsal players with high aerobic capacity using ETM and non-ETM are more effective than male futsal players using ETM and non-ETM. In addition, there is no interaction between groups. Conclusion: This study concluded that Futsal players with aerobic capacity were more effective than those with low aerobic capacity, both for aerobic capacity using ETM and without ETM compared to aerobic capacity using ETM and without ETM. However, the results may differ if there is an increase in the frequency of exercise using the ETM and an increase in the duration of the training period. [ABSTRACT FROM AUTHOR]

Background: Bacillus cereus is a notorious foodborne pathogen, which can grow under anoxic conditions. Anoxic growth is supported by endogenous redox metabolism, for which the thiol redox proteome serves as an interface. Here, we studied the cysteine (Cys) proteome dynamics of B. cereus ATCC 14579 cells grown under fermentative anoxic conditions. We used a quantitative thiol trapping method combined with proteomics profiling. Results: In total, we identified 153 reactive Cys residues in 117 proteins participating in various cellular processes and metabolic pathways, including translation, carbohydrate metabolism, and stress response. Of these reactive Cys, 72 were detected as reduced Cys. The B. cereus Cys proteome evolved during growth both in terms of the number of reduced Cys and the Cys-containing proteins identified, reflecting its growth-phase-dependence. Interestingly, the reduced status of the B. cereus thiol proteome increased during growth, concomitantly to the decrease of extracellular oxidoreduction potential. Conclusions: Taken together, our data show that the B. cereus Cys proteome during unstressed fermentative anaerobic growth is a dynamic entity and provide an important foundation for future redox proteomic studies in B. cereus and other organisms. [ABSTRACT FROM AUTHOR]

Sterols are essential components of the yeast membrane and their synthesis requires oxygen. Yet, Saccharomyces cerevisiae has developed the ability to take up sterols from the medium under anaerobiosis. Here we investigated sterol uptake efficiency and the expression of genes related to sterol import in Saccharomyces and non-Saccharomyces wine yeast species fermenting under anaerobic conditions. The sterol uptake efficiency of 39 strains was evaluated by flow cytometry (with 25-NBD Cholesterol, a fluorescent cholesterol probe introduced in the medium) and we found an important discrepancy between Saccharomyces and non- Saccharomyces wine yeast species that we correlated to a lower final cell population and a lower fermentation rate. A high uptake of sterol was observed in the various Saccharomyces strains. Spot tests performed on 13 of these strains confirmed the differences between Saccharomyces and non-Saccharomyces strains, suggesting that the presence of the sterol uptake transporters AUS1 and PDR11 could cause these discrepancies. Indeed, we could not find any homologue to these genes in the genome of Hanseniaspora uvarum , H. guillermondii, Lachancea thermotolerans, Torulaspora delbreueckii, Metschnikowia pulcherrima , or Starmarella bacillaris species. The specialization of sterol import function for post genome-duplication species may have favored growth under anaerobiosis. [ABSTRACT FROM AUTHOR]

Ancient microbes invented biochemical mechanisms and assembled core metabolic pathways on an anoxic Earth. Molecular oxygen appeared far later, forcing microbes to devise layers of defensive tactics that fend off the destructive actions of both reactive oxygen species (ROS) and oxygen itself. Recent work has pinpointed the enzymes that ROS attack, plus an array of clever protective strategies that abet the well known scavenging systems. Oxygen also directly damages the low-potential metal centers and radical-based mechanisms that optimize anaerobic metabolism; therefore, committed anaerobes have evolved customized tactics that defend these various enzymes from occasional oxygen exposure. Thus a more comprehensive, detailed, and surprising view of oxygen toxicity is coming into view. Reactive oxygen species (ROS) attack iron-dependent enzymes that contemporary microbes inherited from their anoxic ancestors. Microbes that adapted to live in oxic environments evolved layers of defenses against ROS, rather than discarding iron-centered metabolism. Molecular oxygen directly damages the radical species and low-potential metal centers that are critical for optimal anaerobic metabolism, and these injuries may be more important than ROS stress in constraining anaerobes to anoxic environments. Anaerobes periodically confront oxygen, and they have developed esoteric tactics to minimize injuries to some of their most vulnerable enzymes. [ABSTRACT FROM AUTHOR]

The yeast Lachancea thermotolerans is of significant biotechnological interest, and selected strains of this species have become commonly used starter cultures in wine fermentation. However, the impact of this species on wine is frequently limited by the rapid dominance of Saccharomyces cerevisiae strains which are better adapted to wine alcoholic fermentation conditions. Previous studies have shown that the major limiting factor for L. thermotolerans competitive performance in the wine ecosystem is oxygen availability, and not ethanol levels as had been previously suggested. Here we investigated the transcriptional response of L. thermotolerans to anaerobiosis in wine fermentation conditions. The data show that L. thermotolerans broadly redirects gene expression towards genes involved in central carbon metabolism, lipid metabolism, remodeling of the cell wall as well as autophagy. Furthermore, the induction of genes that are likely involved in the generation of lactate indicates a redirection of metabolic flux towards this metabolite. The data provide the first insight into the oxygen-dependent response of L. thermotolerans and suggest potential genetic targets to improve lactate production and/or anaerobic fermentation performance of this yeast. [ABSTRACT FROM AUTHOR]

The Gram-negative bacterium Vibrio cholerae is responsible for the severe diarrheal pandemic disease cholera, representing a major global public health concern. This pathogen transitions from aquatic reservoirs into epidemics in human populations, and has evolved numerous mechanisms to sense this transition in order to appropriately regulate its gene expression for infection. At the intersection of pathogen and host in the gastrointestinal tract lies the community of native gut microbes, the gut microbiome. It is increasingly clear that the diversity of species and biochemical activities within the gut microbiome represents a driver of infection outcome, through their ability to manipulate the signals used by V. cholerae to regulate virulence and fitness in vivo. A better mechanistic understanding of how commensal microbial action interacts with V. cholerae pathogenesis may lead to novel prophylactic and therapeutic interventions for cholera. Here, we review a subset of this burgeoning field of research. [ABSTRACT FROM AUTHOR]

Background: The bacterial genus Exiguobacterium includes several species that inhabit environments with a wide range of temperature, salinity, and pH. This is why the microorganisms from this genus are known generically as polyextremophiles. Several environmental isolates have been explored and characterized for enzyme production as well as for bioremediation purposes. In this line, toxic metal(loid) reduction by these microorganisms represents an approach to decontaminate soluble metal ions via their transformation into less toxic, insoluble derivatives. Microbial-mediated metal(loid) reduction frequently results in the synthesis of nanoscale structures—nanostructures (NS) —. Thus, microorganisms could be used as an ecofriendly way to get NS. Results: We analyzed the tolerance of Exiguobacterium acetylicum MF03, E. aurantiacum MF06, and E. profundum MF08 to Silver (I), gold (III), and tellurium (IV) compounds. Specifically, we explored the ability of cell-free extracts from these bacteria to reduce these toxicants and synthesize NS in vitro, both in the presence or absence of oxygen. All isolates exhibited higher tolerance to these toxicants in anaerobiosis. While in the absence of oxygen they showed high tellurite- and silver-reducing activity at pH 9.0, whereas AuCl4− which was reduced at pH 7.0 in both conditions. Given these results, cell-free extracts were used to synthesize NS containing silver, gold or tellurium, characterizing their size, morphology and chemical composition. Silver and tellurium NS exhibited smaller size under anaerobiosis and their morphology was circular (silver NS), starred (tellurium NS) or amorphous (gold NS). Conclusions: This nanostructure-synthesizing ability makes these isolates interesting candidates to get NS with biotechnological potential. [ABSTRACT FROM AUTHOR]

The microaerophilic human parasite Trichomonas vaginalis causes infections in the urogenital tract and is one of the most often sexually transmitted pathogens worldwide. Due to its anaerobic metabolism, it has to quickly remove intracellular oxygen in order to avoid deactivation of essential metabolic enzymes such as oxygen-sensitive pyruvate:ferredoxin oxidoreductase (PFOR). Two major enzyme activities which are responsible for the removal, i.e. reduction, of molecular oxygen have been identified in T. vaginalis flavin reductase, formerly designated NADPH oxidase, which indirectly reduces oxygen to hydrogen peroxide via flavin mononucleotide (FMN), and NADH oxidase which reduces oxygen to water. Flavin reductase has been identified and characterized at the gene level as well as enzymatically, but NADH oxidase has so far only been characterized enzymatically with enzyme isolated from T. vaginalis cell extracts. In this study, we identified NADH oxidase by mass spectrometry after isolation of the enzyme from gel bands positively staining for NADH oxidase activity. In strain C1 (ATCC 30001) which is known to lack NADH oxidase activity completely, the NADH oxidase gene has a deletion at position 1540 of the open reading frame leading to a frame shift and, as a consequence, to premature termination of the encoded polypeptide. [ABSTRACT FROM AUTHOR]

The adaptation of eukaryotic cells to anaerobic conditions is reflected by substantial changes to mitochondrial metabolism and functional reduction. Hydrogenosomes belong among the most modified mitochondrial derivative and generate molecular hydrogen concomitant with ATP synthesis. The reduction of mitochondria is frequently associated with loss of peroxisomes, which compartmentalize pathways that generate reactive oxygen species (ROS) and thus protect against cellular damage. The biogenesis and function of peroxisomes are tightly coupled with mitochondria. These organelles share fission machinery components, oxidative metabolism pathways, ROS scavenging activities, and some metabolites. The loss of peroxisomes in eukaryotes with reduced mitochondria is thus not unexpected. Surprisingly, we identified peroxisomes in the anaerobic, hydrogenosome-bearing protist Mastigamoeba balamuthi. We found a conserved set of peroxin (Pex) proteins that are required for protein import, peroxisomal growth, and division. Key membrane-associated Pexs (MbPex3, MbPex11, and MbPex14) were visualized in numerous vesicles distinct from hydrogenosomes, the endoplasmic reticulum (ER), and Golgi complex. Proteomic analysis of cellular fractions and prediction of peroxisomal targeting signals (PTS1/PTS2) identified 51 putative peroxisomal matrix proteins. Expression of selected proteins in Saccharomyces cerevisiae revealed specific targeting to peroxisomes. The matrix proteins identified included components of acyl-CoA and carbohydrate metabolism and pyrimidine and CoA biosynthesis, whereas no components related to either ß-oxidation or catalase were present. In conclusion, we identified a subclass of peroxisomes, named "anaerobic" peroxisomes that shift the current paradigm and turn attention to the reductive evolution of peroxisomes in anaerobic organisms. [ABSTRACT FROM AUTHOR]

The central venous minus arterial carbon dioxide pressure to arterial minus central venous oxygen content ratio (Pcv-aCO2/Ca-cvO2) has been proposed as a surrogate for respiratory quotient and an indicator of tissue oxygenation. Some small observational studies have found that a Pcv-aCO2/Ca-cvO2 > 1.4 was associated with hyperlactatemia, oxygen supply dependency, and increased mortality. Moreover, Pcv-aCO2/Ca-cvO2 has been incorporated into algorithms for tissue oxygenation evaluation and resuscitation. However, the evidence for these recommendations is quite limited and of low quality. The goal of this narrative review was to analyze the methodological bases, the pathophysiologic foundations, and the experimental and clinical evidence supporting the use of Pcv-aCO2/Ca-cvO2 as a surrogate for respiratory quotient. Physiologically, the increase in respiratory quotient secondary to critical reductions in oxygen transport is a life-threatening and dramatic event. Nevertheless, this event is easily noticeable and probably does not require further monitoring. Since the beginning of anaerobic metabolism is indicated by the sudden increase in respiratory quotient and the normal range of respiratory quotient is wide, the use of a defined cutoff of 1.4 for Pcv-aCO2/Ca-cvO2 is meaningless. Experimental studies have shown that Pcv-aCO2/Ca-cvO2 is more dependent on factors that modify the dissociation of carbon dioxide from hemoglobin than on respiratory quotient and that respiratory quotient and Pcv-aCO2/Ca-cvO2 may have distinct behaviors. Studies performed in critically ill patients have shown controversial results regarding the ability of Pcv-aCO2/Ca-cvO2 to predict outcome, hyperlactatemia, microvascular abnormalities, and oxygen supply dependency. A randomized controlled trial also showed that Pcv-aCO2/Ca-cvO2 is useless as a goal of resuscitation. Pcv-aCO2/Ca-cvO2 should be carefully interpreted in critically ill patients. [ABSTRACT FROM AUTHOR]

To address the major issue of postharvest losses (20-50%) of a productive crop like tomatoes, a novel aloevera gel based edible coating was developed as postharvest treatment. The objectives of this study were to develop and optimize aloevera gel based edible coating and to study the effect of storage on physicochemical parameters related to tomato quality and safety. For this purpose, an edible coating from aloevera gel was developed, analyzed and applied to tomatoes. There were five treatments (A0, A20, A40, A60, A80) varying in aloevera gel concentration from 0 to 80%. Samples having different percentages of aloevera gel were analyzed for their, physiochemical, textural and microstructural parameters during storage period of 30 days at refrigerated conditions with an interval of 10 days. Control treatment (A0) showed rapid deterioration with an estimated shelf life of 14 days as compared to A80 treatment with extended shelf life of 35 days. Percent weight loss, Size modification, decay percentage, color changes and decrease in firmness was higher for A0 (control) 20, 13, 92, 31.69 and 37%, respectively, whilst the minimum in A80 (Tomatoes coated with 80% aloevera gel) as 4, 0.4, 7.69, 19.73 and 11.46%, accordingly. A significant decrease in acidity value of control tomatoes was observed from 4.56±0.32 to 4.28±0.03 as compared to A80. Microstructural analysis showed that coating tomatoes with coating solution having 40% aloevera gel gave uniformity and continuity on the surface of tomatoes It was concluded that aloevera gel could be an excellent edible coating material and should be used commercially as a technologically viable postharvest preservation technique for fresh produces. [ABSTRACT FROM AUTHOR]

The chemical composition and the presence of microorganisms in liquid silage of peach (Prunus pérsica) were evaluated. A completely randomized design with a 2 * 2 factorial arrangement, four treatments with two percentages of inclusion of molasses and ammonium sulfate was used: T1 (10% molasses - 1% SAM), T2 (10% molasses - 3% SAM), T3 (20% molasses - 1% SAM) and T4 (20% molasses - 2% SAM), with three replications, which were stored between 17 and 19 ° C. The physical-chemical composition of the fruit was determined, taking pH values and microbiological growth, bacterial morphology was characterized on blood agar and MacConkey agar. The organoleptic evaluation of the final silage is established as GOOD according to fermentation characteristics. The protein value increased in each of the treatments as the pH value decreased, associated with high microbial growths, improving the true protein of the food. There was growth of whitish and smooth colonies in blood agar with a predominance of Gram + and Gram - bacillary forms. Pink color - smooth surface colonies grew on MacConkey Agar only on day 0. Liquid peach silage with the addition of SAM increases protein values; and the addition of molasses improves the fermentative parameters such as pH stability. [ABSTRACT FROM AUTHOR]

Animals vary tremendously in their capacities to survive anoxia, and the mechanisms responsible are poorly understood. Adult Drosophila melanogaster are rapidly paralyzed and survive up to 12 h of anoxia, whereas larvae vigorously attempt escape but then die if anoxia exceeds 2 h. Here we use nuclear magnetic resonance methods to compare the metabolome of larvae and adult D. melanogaster under normoxic conditions and after various anoxic durations up to 1 h. Glucose increased during anoxia in both larvae and adults, so anoxic death by carbohydrate limitation is unlikely for either stage. Lactate and alanine were the primary anaerobic end products in both adults and larvae. During the first 30 min of anoxia, larvae accumulated anaerobic end products (predominately lactate) at a higher rate, suggesting that larvae may experience greater initial acid-base disruption during anoxic exposures. Adult Drosophila did not possess higher levels of putative protective metabolites; however, these increased during anoxia in adults and decreased in larvae. Metabolites that decreased during anoxia in larvae included mannitol, xylitol, glycerol, betaine, serine, and tyrosine, perhaps due to use as fuels, antioxidants, or binding to denatured proteins. Adults showed significant increases in glycine, taurine, and the polyols glycerol, mannitol, and xylitol, suggesting that adults upregulate protective metabolites to prevent damage. Our results suggest that lower initial metabolic demand due to paralytic hypometabolism and capacities to upregulate protective metabolites may assist the better anoxia tolerance of adult Drosophila. [ABSTRACT FROM AUTHOR]

Bacterial pathogens such as Salmonella enterica serovar Typhimurium (S. Typhimurium) have to cope with fluctuating oxygen levels during infection within host gastrointestinal tracts. The global transcription factor FNR (fumarate nitrate reduction) plays a vital role in the adaptation of enteric bacteria to the low oxygen environment. Nevertheless, a comprehensive profile of the FNR regulon on the proteome level is still lacking in S. Typhimurium. Herein, we quantitatively profiled S. Typhimurium proteome of an fnr-deletion mutant during anaerobiosis in comparison to its parental strain. Notably, we found that FNR represses the expression of virulence genes of Salmonella pathogenicity island 1 (SPI-1) and negatively regulates propanediol utilization by directly binding to the promoter region of the pdu operon. Importantly, we provided evidence that S. Typhimurium lacking fnr exhibited increased antibiotics susceptibility and membrane permeability as well. Furthermore, genetic deletion of fnr leads to decreased bacterial survival in a Caenorhabditis elegans infection model, highlighting an important role of this regulator in mediating host-pathogen interactions. [ABSTRACT FROM AUTHOR]

Diving optimality models predict air breathers to routinely dive within aerobic limits, but predator avoidance dives may be an exception. Lengthening submergence times during a predation threat may enhance survival probability, and we therefore hypothesized that predator avoidance dives in juvenile estuarine crocodiles (Crocodylus porosus) would be partially anaerobically fueled. We also predicted that reliance on anaerobic metabolism would increase at elevated temperatures to offset the faster depletion of body oxygen stores. Crocodiles were maintained at 287 and 347C for 60 d and subsequently underwent simulated predator avoidance dive trials at two test temperatures (287 and 347C). Blood was sampled immediately on surfacing to measure plasma lactate concentrations relative to nondiving (control) values. Aerobic dive limits (cADL; min) were also calculated using known body mass and oxygen storage relationships and rates of diving oxygen consumption and compared with observed dive durations. Postdive plasma lactate levels were elevated beyond resting levels at both test temperatures, indicating that aerobic thresholds were surpassed during simulated predator avoidance dives. Similarly, ≥90% of dive durations exceeded cADLs at both test temperatures. Postdive plasma lactate concentrations were independent of water temperature and thermal acclimation treatment. Together, these findings suggest that reliance on anaerobiosis during simulated predator avoidance dives is important regardless of temperature. [ABSTRACT FROM AUTHOR]

We describe a collection of aquatic and wetland habitats in an inland landscape, and their occurrence within a terrestrial matrix, as a "freshwater ecosystem mosaic" (FEM). Aquatic and wetland habitats in any FEM can vary widely, from permanently ponded lakes, to ephemerally ponded wetlands, to groundwater‐fed springs, to flowing rivers and streams. The terrestrial matrix can also vary, including in its influence on flows of energy, materials, and organisms among ecosystems. Biota occurring in a specific region are adapted to the unique opportunities and challenges presented by spatial and temporal patterns of habitat types inherent to each FEM. To persist in any given landscape, most species move to recolonize habitats and maintain mixtures of genetic materials. Species also connect habitats through time if they possess needed morphological, physiological, or behavioral traits to persist in a habitat through periods of unfavorable environmental conditions. By examining key spatial and temporal patterns underlying FEMs, and species‐specific adaptations to these patterns, a better understanding of the structural and functional connectivity of a landscape can be obtained. Fully including aquatic, wetland, and terrestrial habitats in FEMs facilitates adoption of the next generation of individual‐based models that integrate the principles of population, community, and ecosystem ecology. Research Impact Statement: Fully including aquatic, wetland, and terrestrial habitats facilitates adoption of next‐generation, individual‐based, models that integrate principles of population, community, and ecosystem ecology. [ABSTRACT FROM AUTHOR]

BACKGROUND Lactic acid bacteria inocula have been developed over the years to improve the aerobic stability of silages. The aims of the study were to evaluate the effect of various conservation periods and the use of Lactobacillus hilgardii inoculum on aerobic stability, fermentative profile and microbial population of corn and sorghum silages. Trials were carried out on two corns and one sorghum crops. The crops were untreated or treated with L. buchneri (LB, application rate 300 000 cfu g−1 FM), L. hilgardii (LH, application rate 300 000 cfu g−1 FM) and a combination (LB + LH, application rate 150 000 cfu g−1 FM each). Silos were opened after 15, 30, 100 and 250 days of conservation, and the silages were analysed for fermentative profile, microbial count and aerobic stability. RESULTS: During conservation, the inocula influenced the fermentation profile. The use of LH increased the aerobic stability at 15 and 30 days in one out of three trials, while after 100 and 250 days, the presence of LB alone or with LH led to greater stability. In all the trials, the acetic acid content increased, the yeast count decreased and the aerobic stability increased during the conservation period. CONCLUSIONS: This study has shown that a long period in complete anaerobiosis reduced yeast count and improved aerobic stability in all silages. The addition of LB was confirmed to be a good option for increasing aerobic stability of silages, whereas the effect of LH alone or in combination with LB on aerobic stability was not consistent between trials. © 2018 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The six-component maintenance of lipid asymmetry (Mla) system is responsible for retrograde transport of phospholipids, ensuring the barrier function of the Gram-negative cell envelope. Located within the outer membrane, MlaA (VacJ) acts as a channel to shuttle phospholipids from the outer leaflet. We identified Neisseria gonorrhoeae MlaA (ngo2121) during high-throughput proteomic mining for potential therapeutic targets against this medically important human pathogen. Our follow-up phenotypic microarrays revealed that lack of MlaA results in a complex sensitivity phenome. Herein we focused on MlaA function in cell envelope biogenesis and pathogenesis. We demonstrate the existence of two MlaA classes among 21 bacterial species, characterized by the presence or lack of a lipoprotein signal peptide. Purified truncated N. gonorrhoeae MlaA elicited antibodies that cross-reacted with a panel of different Neisseria. Little is known about MlaA expression; we provide the first evidence that MlaA levels increase in stationary phase and under anaerobiosis but decrease during iron starvation. Lack of MlaA resulted in higher cell counts during conditions mimicking different host niches; however, it also significantly decreased colony size. Antimicrobial peptides such as polymyxin B exacerbated the size difference while human defensin was detrimental to mutant viability. Consistent with the proposed role of MlaA in vesicle biogenesis, the ΔmlaA mutant released 1.7-fold more membrane vesicles. Comparative proteomics of cell envelopes and native membrane vesicles derived from ΔmlaA and wild type bacteria revealed enrichment of TadA–which recodes proteins through mRNA editing–as well as increased levels of adhesins and virulence factors. MlaA-deficient gonococci significantly outcompeted (up to 16-fold) wild-type bacteria in the murine lower genital tract, suggesting the growth advantage or increased expression of virulence factors afforded by inactivation of mlaA is advantageous in vivo. Based on these results, we propose N. gonorrhoeae restricts MlaA levels to modulate cell envelope homeostasis and fine-tune virulence. [ABSTRACT FROM AUTHOR]

Osm1, a soluble fumarate reductase from Saccharomyces cerevisiae, is localized in both the mitochondria and the endoplasmic reticulum (ER). OSM1 genetically interacts with ERO1, which encodes an essential ER oxidoreductase for disulfide‐bond formation under anaerobic conditions. However, the detailed enzymatic mechanisms involved in this interaction and the cellular roles of Osm1 are not fully understood. In this study, monomeric and stable recombinant Osm1 was successfully prepared for structural study. During purification, it was realized that the majority of recombinant Osm1 expressed in Escherichia coli lacked the flavin adenosine dinucleotide (FAD) cofactor. However, exogenously introduced FAD could be incorporated into recombinant Osm1, generating stable and homogenous holo Osm1. Moreover, after removing a flexible fragment by limited proteolysis, holo Osm1 formed isotropic crystals that retained catalytic activity. X‐ray diffraction data were successfully collected from the Osm1 crystals to a resolution of 1.75 Å. Osm1, a soluble fumarate reductase from Saccharomyces cerevisiae, was purified and crystallized. The crystals were found to belong to space group P21, with unit‐cell parameters a = 43.88, b = 109.32, c = 49.93 Å, β = 112.25°. Crystals were obtained at 293 K and diffracted to a resolution of 1.75 Å. [ABSTRACT FROM AUTHOR]

Abstract Low yield and inhibition of hydrogenase by oxygen are the main limitations for hydrogen production by microalgae. Considering the role of electron flow in the metabolism for hydrogen production, a genome-scale metabolic model (named i MM627) was reconstructed for Auxenochlorella protothecoides. i MM627 was evaluated using experimental data for growth and flux distribution. Then, considering the well-known degeneracy of FBA solutions, a new method of finding effective reactions based on multiple optimal solutions was developed. At a constant growth rate, flux distributions for maximal and minimal hydrogen production under anaerobiosis and for maximal oxygen production were compared to identify target reactions for improvement of hydrogen production and for providing anaerobiosis. Existing researches on some reactions truly confirm the predicted changes by i MM627. The main proposed strategies for improvement of hydrogen production include changes in metabolism to provide NADH. Consumption of oxygen by mitochondrial respiration and energy dissipation was proposed to provide anaerobiosis. Graphical abstract Image 1 Highlights • The first genome-scale metabolic model was reconstructed for A. protothecoides. • The model was employed as a framework to study hydrogen and oxygen production. • A new method of finding effective reactions was developed by optimal solutions. • The method was applied to find strategies for improvement of hydrogen production. [ABSTRACT FROM AUTHOR]

We report the discovery of a new genus of heterolobosean flagellates, Dactylomonas gen. nov., with two species, D. venusta sp. nov. and D. crassa sp. nov. Phylogenetic analysis of the SSU rRNA gene showed that Dactylomonas is closely related to the amoeba Selenaion, the deepest‐branching lineage of Tetramitia. Dactylomonads possess two flagella, and ultrastructural studies revealed an unexpected organization of the flagellar apparatus, which resembled Pharyngomonada (the second lineage of Heterolobosea) instead of Tetramitia: basal bodies were orthogonal to each other and a putative root R1 was present in the mastigont. On the other hand, Dactylomonas displayed several features uncommon in Heterolobosea: a microtubular corset, a distinctive rostrum supported by the main part of the right microtubular root, a finger‐like projection on the proximal part of the recurrent flagellum, and absence of a ventral groove. In addition, Dactylomonas is anaerobic and seems to have lost mitochondrial cristae. Dactylomonas and Selenaion are accommodated in the family Selenaionidae fam. nov. and order Selenionida ord. nov. The taxonomy of Tetramitia is partially revised, and the family Neovahlkampfiidae fam. nov. is established. [ABSTRACT FROM AUTHOR]

This study analyses the induction and repression of nitrate reduction activity in a batch culture of Idiomarina strain cos21. On a change from aerobic to anaerobic respiration, the culture entered a stationary phase. The onset of this phase showed 3.75 fold increase in mRNA levels for the nitrate reductase enzyme. mRNA accumulated very rapidly during a short period, after which its overall concentration declined to reach a lower value. The level of nitrite reductase protein reached a maximum value at 36 h of growth when the oxygen concentration dropped below 10 µM. The data set provided here confer new insights into the understanding of the physiological response of Idiomarina strain cos21 to change in oxygen concentration allowing the bacterium to survive and adapt to a new environment by dissimilatory reduction of nitrate to nitrite, which serves to provide energy as the bacteria adapt to anaerobiosis. Main strategy used here is to induce, measure, and track the expression of microbial genes, while they grow in culture conditions to better mimic interaction in a natural environment. This study will help us with a better understanding of the nitrate reduction process in the oxygen minimum zone. [ABSTRACT FROM AUTHOR]

In Euglena gracilis, wax ester fermentation produces ATP during anaerobiosis. Here, we report that anaerobic wax ester production is suppressed when the mitochondrial electron transport chain complex I is inhibited by rotenone, whereas it is increased by the uncoupler carbonyl cyanide m‐chlorophenylhydrazone (CCCP). The ADP/ATP ratio in anaerobic cells is elevated by treatment with either rotenone or CCCP. Gene silencing experiments indicate that acyl‐CoA dehydrogenase, electron transfer flavoprotein (ETF), and rhodoquinone (RQ) participate in wax ester production. These results suggest that fatty acids are synthesized in mitochondria by the reversal of β‐oxidation, where trans‐2‐enoyl‐CoA is reduced mainly by acyl‐CoA dehydrogenase using the electrons provided by NADH via the electron transport chain complex I, RQ, and ETF, and that ATP production is highly supported by anaerobic respiration utilizing trans‐2‐enoyl‐CoA as a terminal electron acceptor. [ABSTRACT FROM AUTHOR]

Abstract In the production of fuel ethanol from sugarcane in Brazilian biorefineries, one way to achieve lower production costs is the fermentation of sugars into ethanol at high temperatures (˜48 °C), which could lead to a decrease in: contamination levels, use of antibiotics, cooling costs, water usage, use of sulphuric acid in cell recycling, and energy used in distillation. Since Saccharomyces cerevisiae , the yeast species used in these biorefineries, cannot grow at these temperatures, alternative organisms need to be employed and one candidate is Kluyveromyces marxianus. Here, 20 strains belonging to this species were screened for their capacity to thrive at high temperatures (37, 45 and 48 °C) and under conditions relevant in the fuel ethanol industry: low pH (2.5), anaerobiosis, and high ethanol concentrations (up to 10%). The ethanol producing capacity of some strains was also evaluated and the ethanol yield on sugar quantified. Strains K. marxianus NCYC 3396 and UFV-3 stood out as potential microorganisms to be plugged into this industry (metabolic, evolutionary and reverse engineering will be required), since they grew better than the remaining ones under most of the conditions tested and produced ethanol at 48 °C with similar yields (0.40 g ethanol.g glucose−1) to those displayed by S. cerevisiae at 37 °C. Highlights • Four K. marxianus strains have potential to thrive in the presence of stressors found in the fuel ethanol industry in Brazil. • K. marxianus NCYC 3396 and UFV-3 were capable of growing anaerobically in complex medium. • K. marxianus NCYC 3396 and UFV-3 yielded 0.40 g ethanol.g glucose−1 at 48 °C in a synthetic medium. • Cell viability was severely jeopardized when K. marxianus UFV-3 was tested in a miniaturized industrial system above 40 °C. [ABSTRACT FROM AUTHOR]

Next generation biofuels including longer-chain alcohols such as butanol are attractive as renewable, high-energy fuels. A barrier to microbial production of butanols is the increased toxicity compared to ethanol; however, the cellular targets and microbial defense mechanisms remain poorly understood, especially under anaerobic conditions used frequently in industry. Here we took a comparative approach to understand the response of Saccharomyces cerevisiae to 1-butanol, isobutanol, or ethanol, across three genetic backgrounds of varying tolerance in aerobic and anaerobic conditions. We find that strains have different growth properties and alcohol tolerances with and without oxygen availability, as well as unique and common responses to each of the three alcohols. Our results provide evidence for strain-byalcohol- by-oxygen interactions that moderate how cells respond to alcohol stress. [ABSTRACT FROM AUTHOR]

Ocean warming and acidification are two important environmental drivers affecting marine organisms. Organisms living at high latitudes might be especially threatened in near future, as current environmental changes are larger and occur faster. Therefore, we investigated the effect of hypercapnia on thermal tolerance and physiological performance of sub-Arctic Mytilus edulis from the White Sea. Mussels were exposed (2 weeks) to 390 µatm (control) and 1120 µatm CO2 (year 2100) before respiration rate (MO2), anaerobic metabolite (succinate) level, haemolymph acid-base status and intracellular pH (pHi) were determined during acute warming (10-28 °C, 3 °C over night). In normocapnic mussels, warming induced MO2 to rise exponentially until it levelled off beyond a breakpoint temperature of 20.5 °C. Concurrently, haemolymph PCO2 rose significantly > 19 °C followed by a decrease in PO2 indicating the pejus temperature (TP, onset of thermal limitation). Succinate started to accumulate at 28 °C under normocapnia defining the critical temperature (TC). pHi was maintained during warming until it dropped at 28 °C, in line with the concomitant transition to anaerobiosis. At acclimation temperature, CO2 had only a minor impact. During warming, MO2 was stimulated by CO2 resulting in an elevated breakpoint of 25.8 °C. Nevertheless, alterations in haemolymph gases (> 16 °C) and the concomitant changes of pHi and succinate level (25 °C) occurred at lower temperature under hypercapnia versus normocapnia indicating a downward shift of both thermal limits TP and TC by CO2. Compared to temperate conspecifics, sub-Arctic mussels showed an enhanced thermal sensitivity, exacerbated further by hypercapnia, indicating their potential vulnerability to environmental changes projected for 2100. [ABSTRACT FROM AUTHOR]

The African clawed frog, Xenopus laevis , can withstand extremely arid conditions through aestivation, resulting in dehydration and urea accumulation. Aestivating X. laevis reduce their metabolic rate, and rely on anaerobic glycolysis to meet reduced ATP demands. The present study investigated how severe dehydration affected the transcript levels, kinetic profile, and phosphorylation state of the key glycolytic enzyme pyruvate kinase (PK) in the liver and skeletal muscle of X. laevis . Compared to control frogs, severely dehydrated frogs showed an increase in the transcript abundance of both liver and muscle isoforms of PK. While the kinetics of muscle PK did not differ between dehydrated and control frogs, PK from the liver of dehydrated frogs had a lower K m for phosphoenolpyruvate (PEP) (38%), a lower K a for fructose-1,6-bisphosphate (F1,6P 2 ) (32%), and a greater activation of PK via F1,6P 2 (1.56-fold). PK from dehydrated frogs also had a lower phosphorylation-state (25%) in comparison to the enzyme from control frogs in the liver. Experimental manipulation of the phosphorylation-state of liver PK taken from control frogs by endogenous protein phosphatases resulted in decreased phosphorylation, and a similar kinetic profile as seen in dehydrated frogs. The physiological consequence of dehydration-induced PK modification appears to adjust PK function to remain active during a metabolically depressed state. This study provides evidence for the maintenance of PK activity through elevated mRNA levels and a dephosphorylation event which activates frog liver PK in the dehydrated state in order to facilitate the production of ATP via anaerobic glycolysis. [ABSTRACT FROM AUTHOR]

Hypoxia in the root zone activates several mechanisms to cope with the energetic imbalance that triggers hypoxia. Prunus is a hypoxia-sensitive species, and there is not information about the relationship between components of the root respiration and the hypoxia tolerance. We investigated whether adjusting root respiratory components plays a role in the hypoxia tolerance in Prunus rootstocks genotypes. Three Prunus rootstocks genotypes, ‘Mariana 2624’ (tolerant), ‘CAB6P’ (moderately sensitive) and ‘Mazzard F12/1’ (sensitive), were exposed to normoxia and hypoxia in a hydroponic system for 30 days. During the first 10 days of the experiment, the hypoxia-tolerant genotype under hypoxia exhibited a higher root relative growth rate, maintained net nitrogen uptake rate, and had lower protein turnover and electrolyte leakage, as well as increased root porosity (aerenchyma formation from 0.3 to 6.0 cm from the root tip at 30 days of experiment) compared to hypoxia-sensitive genotypes. ‘Mariana 2624’ roots exhibited tolerance to hypoxia by reducing maintenance respiration by 50% compared to normoxia, while growth respiration increased. Control of root respiratory components, early response of ion uptake favors root relative growth rate, and change of the root anatomy are associated with the ability to tolerate hypoxia by reducing maintenance costs. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • The chemical family distribution of VOC in pile was homogeneous during composting. • Concentrations of emitted VOC was not dependent on their concentrations in the pile. • Amount and chemical distribution of emitted VOC were driven by microbial activity. • Anaerobic conditions altered the chemical family distribution of emitted VOC. Abstract Composting operations taking place at municipal solid waste (MSW) treatment plants represent a source of volatile organic compounds (VOC) to the atmosphere. Understanding the variables governing the release of VOC at these facilities is crucial to assess potential health risks for site workers and local residents. In this work the changes in the VOC composition of a composting pile were monitored and compared to the VOC emmited from the same pile in order to understand the impact of composting operations on the release of VOC. More than one hundred VOC were indentified in the solid phase of the composting piles, which were dominated by terpenes (about 50% of the total amount of VOC) and in a lower quantity alcohols, volatile fatty acids and aromatic compounds. There was a reduction in the total concentration of VOC in the pile during composting, from 45 to 35 mg/kg, but the compostion and distribution of VOC families remained stable in the pile even in the mature compost. However, there was no correlation between the emitted VOC and their concentration in the composting pile. The VOC emission pattern was affected by the biological activity in the pile (measured by temperature, CO 2 evolution and the presence of CH 4 emissions). The highest VOC emissions were detected at early stages of the process, alongside with the generation of CH 4 in the pile, and then decreased sharply in the mature compost as a consequence of biodegradation and volatilisation. These results pointed to the importance of composting operation rather than the composition of the raw materials on the release of VOC in composting plants. [ABSTRACT FROM AUTHOR]

Various packaging technologies have been developed to meet continuously increasing consumer demand for comfortable and safe use, convenience of use, and fresh quality of packaged products. Porous and breathable films provide various advantages such as the minimization of anaerobiosis originating from the accumulation of CO 2 in packaging and the automatic release of steam in a microwave system. Non-porous breathable films are among the alternative candidates for packaging applications and they can respond to desired and undesired temperature fluctuations by reversibly changing the gas permeability depending on the temperature change. They can also prevent problems related to packaging integrity such as the intrusion of insects and microorganisms through perforations, undesired permeation of moisture, and loss of flavor in agricultural products and food. This review mainly addresses porous and non-porous breathable films that are responsive to stimuli like temperature changes. Further, the importance and possible packaging application areas of non-porous breathable films with temperature-dependent gas permeabilities are briefly identified. [ABSTRACT FROM AUTHOR]

Although the concept of ensiling large-round or large-square bales dates back to the late 1970s, many refinements have been made to both equipment and management since that time, resulting in much greater acceptance by small or mid-sized dairy or beef producers. This silage preservation technique is attractive to producers for several reasons, but the primary advantage is a reduced risk of weather damage to valuable forage crops compared with preservation as dry hay. Most core principles for making high-quality precisionchopped silages also apply to baled silages; among these, establishing and subsequently maintaining anaerobiosis are priorities. For baled silages, these priorities are critical, in part because recommended moisture concentrations (45 to 55%) are drier, and particle length is much longer. These factors act to restrict the rate and extent of silage fermentation, often resulting in less production of desirable fermentation acids and a greater (less acidic) final pH. Within this context, preservation of baled silages can be improved by applying polyethylene (PE) film wraps promptly, using an appropriate number of PE film layers (6 to 8), selecting a storage site free of sharp objects or other debris, and by monitoring wrapped bales closely for evidence of puncture, particularly by birds or vermin. Under certain conditions, such as those in which the bale moisture of highly buffered forages exceeds the recommended range, the heterogeneous nature of baled silages coupled with a restricted rate and extent of fermentation may increase susceptibility to clostridial activity compared with precision- chopped forages ensiled at comparable moisture concentrations. To date, research evaluating inoculants or other additives designed to improve the fermentation of challenging forages or aerobic stability has been limited, but should not be discontinued. Development of PE film embedded with an oxygen-limiting barrier has yielded positive results in some trials; however, most differences between these novel formulations and reputable commercial PE film have been related to decreases in yeast and mold counts at the surface layer. Related assessments of fermentation or nutritive value determined on a whole-bale basis have been less conclusive. Baled silages can be produced successfully by adhering to straightforward management principles; as such, this form of silage production is likely to remain popular for the foreseeable future. [ABSTRACT FROM AUTHOR]