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13 pages, 1596 KiB  
Article
Nitrogen Fixation, Carbohydrate Contents, and Bacterial Microbiota in Unelongated Stem of Manure Compost-Applied Rice at Panicle Initiation
by Zhalaga Ao, Miu Tsuchiya, Juan Xia, Chie Hayakawa, Yukitsugu Takahashi, Hideaki Hirai and Isamu Maeda
Microbiol. Res. 2024, 15(3), 1900-1912; https://doi.org/10.3390/microbiolres15030127 (registering DOI) - 15 Sep 2024
Viewed by 180
Abstract
In rice, symbiotic N2 fixation via nodule bacteroids does not take place naturally. Although N2 fixation by endophytic and associative diazotrophs has been reported in rice, the main organs and seasonal regulation for the N2 fixation have not been elucidated. [...] Read more.
In rice, symbiotic N2 fixation via nodule bacteroids does not take place naturally. Although N2 fixation by endophytic and associative diazotrophs has been reported in rice, the main organs and seasonal regulation for the N2 fixation have not been elucidated. In this study, seasonal changes in nitrogenase (acetylene reduction) activity and carbohydrate contents in elongated culm (EC), unelongated stem (US), and crown root (CR) were investigated in manure compost (MC)- and chemical fertilizer (CF)-applied rice. Nitrogenase activity increased after rooting (June) and reached the highest activity in US of MC-applied rice at panicle initiation (August). The sucrose content in EC continued to increase after rooting regardless of the applied materials, whereas the glucose content in US increased after rooting only in CF-applied rice, suggesting higher consumption of glucose in US of MC-applied rice. There were significant differences among bacterial microbiota in EC, US, and CR at panicle initiation. In addition, Clostridia class anaerobes were more abundant in US of MC-applied rice than in EC and CR of MC-applied rice. Such difference was not observed in US of CF-applied rice. These results suggest the suitability of US of MC-applied rice at panicle initiation as a site of N2 fixation under anaerobic conditions. Full article
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<p>The parts of rice cultured in a wetland rice field at Utsunomiya University, Moka, Japan, and collected at panicle initiation. The indicated parts were cut from the whole plant (<b>A</b>). The rice parts from chemical fertilizer (CF)-applied rice (<b>B</b>) and manure compost (MC)-applied rice (<b>C</b>) are shown. The centimeter scale is shown.</p>
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<p>Properties related to nitrogen fixation in elongated culm (EC), unelongated stem (US), and crown root (CR) of manure compost (MC)- and chemical fertilizer (CF)-applied rice at panicle initiation in 2022. Columns with error bars for nitrogenase activity (<b>A</b>), <span class="html-italic">nifH</span> copy number/16S rRNA gene copy number (<b>B</b>), sucrose content (<b>C</b>), and glucose content (<b>D</b>) indicate means ± SD (<span class="html-italic">n</span> = 3). Different lowercase letters show significant differences in multiple comparisons among different material-applied rice specimens in different parts (<span class="html-italic">p</span> &lt; 0.05). ND = not detected.</p>
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<p>The principal coordinate analysis (PCoA) of the bacterial microbiota in the parts of manure compost (MC)-applied and chemical fertilizer (CF)-applied rice at panicle initiation. Coordinate points for elongated culm (EC; green), unelongated stem (US; blue), and crown root (CR; red) of MC-applied (closed) and CF-applied (open) rice are shown. Drawings indicate 95% confidence ellipses for EC (green), US (blue), and CR (red) of MC-applied (solid) and CF-applied (dashed) rice.</p>
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<p>Class-level and species-level taxonomic profiles in elongated culm (EC), unelongated stem (US), and crown root (CR) of manure compost (MC)- and chemical fertilizer (CF)-applied rice at panicle initiation. A divided bar chart with error bars is composed of columns showing class-level relative abundance + SD (<span class="html-italic">n</span> = 3) (<b>A</b>). Columns with error bars indicate means of relative abundance for <span class="html-italic">Clostridiales</span> species ± SD (<span class="html-italic">n</span> = 3) (<b>B</b>). Different lowercase letters show significant difference in multiple comparisons among the relative taxonomic abundances in different rice parts of different material-applied rice (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Properties related to nitrogen fixation in elongated culm (EC), unelongated stem (US), and crown root (CR) of manure compost (MC)- and chemical fertilizer (CF)-applied rice at panicle initiation in 2023. Columns with error bars for nitrogenase activity (<b>A</b>), <span class="html-italic">nifH</span> copy number/16S rRNA gene copy number (<b>B</b>), sucrose content (<b>C</b>), and glucose content (<b>D</b>) indicate means ± SD (<span class="html-italic">n</span> = 5). Different lowercase letters show significant differences in multiple comparisons among different material-applied rice specimens in different parts (<span class="html-italic">p</span> &lt; 0.05). ND = not detected.</p>
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21 pages, 814 KiB  
Review
Impacts of Polyvinyl Alcohol and Chitosan-Modified Biochar on the Anaerobic Digestion of Sewage Sludge and Valuable Resource Recovery
by Ping Fa Chiang, Teng Ling Zhang, Ndungutse Jean Maurice, Mugabekazi Joie Claire, Bigirimana Gentil, Abdul Ghaffar Memon and Abdulmoseen Segun Giwa
Processes 2024, 12(9), 1987; https://doi.org/10.3390/pr12091987 - 14 Sep 2024
Viewed by 330
Abstract
The accumulation of organic dyes and heavy metals (HMs) in sewage sludge (SS) after wastewater treatment is a significant problem due to the non-degradable nature of these pollutants. Moreover, the simultaneous removal of HMs and dyes in the complex process of SS treatment, [...] Read more.
The accumulation of organic dyes and heavy metals (HMs) in sewage sludge (SS) after wastewater treatment is a significant problem due to the non-degradable nature of these pollutants. Moreover, the simultaneous removal of HMs and dyes in the complex process of SS treatment, such as anaerobic digestion (AD), has become attractive. HMs and dyes present in SS can have a detrimental effect on anaerobic digesters. These pollutants not only inhibit the production of methane, which is crucial for biogas generation, but also affect the stability of AD treatment, which can result in failure or inadequate performance of the AD process. This review highlights a novel method of removing HMs and dyes from the AD process of SS through the use of biochar modified with polyvinyl alcohol (PVA) and chitosan (CTS). The applications of conventional biochar have been limited due to poor adsorption capacity. However, modification using PVA/CTS composites enhances properties such as surface functional groups, adsorption capacity, porosity, surface area selectivity, and stability. Furthermore, this modified version can function as an additive in AD of SS treatment to boost biogas production, which is a viable source for heat generation or electricity supply. In addition, the digestates can be further processed through plasma pyrolysis for the removal of HMs and dyes bound to the modified biochar. Plasma pyrolysis generates two major products: syngas and slag. The syngas produced can then be used as a source of hydrogen, heat, and electricity, while the slag can potentially be reused as an AD additive or as a biofertilizer in the agricultural sector. Additionally, this study addresses the challenges associated with this integration and biochar modifications, and offers an outlook on understanding the interactions between the modified biochar properties, microbial dynamics, and the presence of micropollutants to ensure the economic viability and scalability of this technology. This comprehensive review provides insights into the potential of PVA/CTS-modified biochar as an effective additive in AD systems, offering a sustainable approach to SS treatment and valuable resource recovery. Full article
(This article belongs to the Special Issue Current Trends in Anaerobic Digestion Processes, 2nd Edition)
13 pages, 5151 KiB  
Article
Direct Conversion of Minimally Pretreated Corncob by Enzyme-Intensified Microbial Consortia
by Alei Geng, Nana Li, Anaiza Zayas-Garriga, Rongrong Xie, Daochen Zhu and Jianzhong Sun
Agriculture 2024, 14(9), 1610; https://doi.org/10.3390/agriculture14091610 - 14 Sep 2024
Viewed by 242
Abstract
The presence of diverse carbohydrate-active enzymes (CAZymes) is crucial for the direct bioconversion of lignocellulose. In this study, various anaerobic microbial consortia were employed for the degradation of 10 g/L of minimally pretreated corncob. The involvement of lactic acid bacteria (LAB) and a [...] Read more.
The presence of diverse carbohydrate-active enzymes (CAZymes) is crucial for the direct bioconversion of lignocellulose. In this study, various anaerobic microbial consortia were employed for the degradation of 10 g/L of minimally pretreated corncob. The involvement of lactic acid bacteria (LAB) and a CAZyme-rich bacterium (Bacteroides cellulosilyticus or Paenibacillus lautus) significantly enhanced the lactic acid production by Ruminiclostridium cellulolyticum from 0.74 to 2.67 g/L (p < 0.01), with a polysaccharide conversion of 67.6%. The supplement of a commercial cellulase cocktail, CTec 2, into the microbial consortia continuously promoted the lactic acid production to up to 3.35 g/L, with a polysaccharide conversion of 80.6%. Enzymatic assays, scanning electron microscopy, and Fourier transform infrared spectroscopy revealed the substantial functions of these CAZyme-rich consortia in partially increasing enzyme activities, altering the surface structure of biomass, and facilitating substrate decomposition. These results suggested that CAZyme-intensified consortia could significantly improve the levels of bioconversion of lignocellulose. Our work might shed new light on the construction of intensified microbial consortia for direct conversion of lignocellulose. Full article
(This article belongs to the Section Agricultural Technology)
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<p>Chemical productions on 10 g/L minimally pretreated corncob by a series of microbial consortia after 3 days of incubation. For the species composition of the consortia, refer to <a href="#agriculture-14-01610-t001" class="html-table">Table 1</a>. Control indicates the chemical composition after inoculation but without incubation. Values are the mean of three replicates. Error bars indicate standard deviation.</p>
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<p>Heatmap of the extracellular enzyme activities of different microbial consortia. <sup>a</sup> Crude enzymes from the culture grown on wheat bran and <sup>b</sup> crude enzymes from the culture grown on corncob. Data were normalized in a row. For the species composition of the consortia, refer to <a href="#agriculture-14-01610-t001" class="html-table">Table 1</a>. Values are the mean of three replicates.</p>
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<p>SEM of the corncob before and after treatment by a series of microbial consortia. (<b>A</b>) Blank control; (<b>B</b>) consortium A (<span class="html-italic">R. cellulolyticum</span> alone); (<b>C</b>) consortium C; (<b>D</b>) consortium D; (<b>E</b>) consortium E; and (<b>F</b>) consortium F. For the species composition of the consortia, refer to <a href="#agriculture-14-01610-t001" class="html-table">Table 1</a>. The images were taken at 2.0 kv, the magnification was set at 2000 times, and the scale bars in the insertion are 4 μm.</p>
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<p>FTIR analysis of the corncob before and after treatment by the microbial consortia. Letters on the graph refer to the treatment by the specific consortia. The assignment of the bands is shown in <a href="#app1-agriculture-14-01610" class="html-app">Table S2</a>.</p>
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<p>Microbial population structures of different consortia during various time spans. (<b>A</b>) At the start point; (<b>B</b>) after 1 day; (<b>C</b>) after 2 days; and (<b>D</b>) after 3 days of incubation (end of fermentation). Letters on the horizontal axis refer to the specific consortia, and different species are distinguished by color.</p>
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<p>Chemical productions on 10 g/L minimally pretreated corncob by enzyme-enhanced microbial consortia after 3 days of incubation. For the species composition of the consortia, refer to <a href="#agriculture-14-01610-t001" class="html-table">Table 1</a>. Values are the mean of three replicates. Error bars indicate standard deviation.</p>
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10 pages, 1955 KiB  
Brief Report
A Metagenomics Pipeline to Characterize Self-Collected Vaginal Microbiome Samples
by Krystal Thomas-White, Evann E. Hilt, Genevieve Olmschenk, Maryann Gong, Caleb D. Phillips, Courtney Jarvis, Nicholas Sanford, Jennifer White and Pita Navarro
Diagnostics 2024, 14(18), 2039; https://doi.org/10.3390/diagnostics14182039 - 13 Sep 2024
Viewed by 390
Abstract
Vaginitis is a widespread issue for women worldwide, yet current diagnostic tools are lacking. Bacterial vaginosis (BV) is the most prevalent type of vaginitis, found in 10–50% of reproductive-aged women. Current diagnostic methods for BV rely on clinical criteria, microscopy, or the detection [...] Read more.
Vaginitis is a widespread issue for women worldwide, yet current diagnostic tools are lacking. Bacterial vaginosis (BV) is the most prevalent type of vaginitis, found in 10–50% of reproductive-aged women. Current diagnostic methods for BV rely on clinical criteria, microscopy, or the detection of a few microbes by qPCR. However, many vaginal infections lack a single etiological agent and are characterized by changes in the vaginal microbiome community structure (e.g., BV is defined as a loss of protective lactobacilli resulting in an overgrowth of anaerobic bacteria). Shotgun metagenomic sequencing provides a comprehensive view of all the organisms present in the vaginal microbiome (VMB), allowing for a better understanding of all potential etiologies. Here, we describe a robust VMB metagenomics sequencing test with a sensitivity of 93.1%, a specificity of 90%, a negative predictive value of 93.4%, and a positive predictive value of 89.6% certified by Clinical Laboratory Improvement Amendments (CLIA), the College of American Pathologist (CAP), and the Clinical Laboratory Evaluation Program (CLEP). We sequenced over 7000 human vaginal samples with this pipeline and described general findings and comparisons to US census data. Full article
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<p>The Evvy VMB test workflow.</p>
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<p>Shotgun metagenomics resolution of mock community samples: (<b>A</b>) Of 1:10 dilution of mock community performed in the wet lab and (<b>B</b>) an in-silico subsampling of the initial mock community sample down to a 4-log decrease.</p>
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<p>Relative abundance graphs of vaginal samples. These profiles are example profiles containing the top 10 species detected by frequency and relative abundance (<a href="#diagnostics-14-02039-t002" class="html-table">Table 2</a>).</p>
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<p>Comparison of Evvy’s user data to the 2020 US census data. Percent of samples stratified by (<b>A</b>) age, or (<b>B</b>) self-reported race and ethnicity compared to reported US census data. * The follow race/ethnicity options were shortened for this figure: Black or African American, American Indian or Alaskan Native, Native Hawaiian or Pacific Islander.</p>
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21 pages, 7200 KiB  
Article
Thermophilic Hemicellulases Secreted by Microbial Consortia Selected from an Anaerobic Digester
by Luca Bombardi, Marco Orlando, Martina Aulitto and Salvatore Fusco
Int. J. Mol. Sci. 2024, 25(18), 9887; https://doi.org/10.3390/ijms25189887 - 13 Sep 2024
Viewed by 540
Abstract
The rise of agro-industrial activities over recent decades has exponentially increased lignocellulose biomasses (LCB) production. LCB serves as a cost-effective source for fermentable sugars and other renewable chemicals. This study explores the use of microbial consortia, particularly thermophilic consortia, for LCB deconstruction. Thermophiles [...] Read more.
The rise of agro-industrial activities over recent decades has exponentially increased lignocellulose biomasses (LCB) production. LCB serves as a cost-effective source for fermentable sugars and other renewable chemicals. This study explores the use of microbial consortia, particularly thermophilic consortia, for LCB deconstruction. Thermophiles produce stable enzymes that retain activity under industrial conditions, presenting a promising approach for LCB conversion. This research focused on two microbial consortia (i.e., microbiomes) that were analyzed for enzyme production using a cheap medium, i.e., a mixture of spent mushroom substrate (SMS) and digestate. The secreted xylanolytic enzymes were characterized in terms of temperature and pH optima, thermal stability, and hydrolysis products from LCB-derived polysaccharides. These enzymes showed optimal activity aligning with common biorefinery conditions and outperformed a formulated enzyme mixture in thermostability tests in the digestate. Phylogenetic and genomic analyses highlighted the genetic diversity and metabolic potential of these microbiomes. Bacillus licheniformis was identified as a key species, with two distinct strains contributing to enzyme production. The presence of specific glycoside hydrolases involved in the cellulose and hemicellulose degradation underscores these consortia’s capacity for efficient LCB conversion. These findings highlight the potential of thermophilic microbiomes, isolated from an industrial environment, as a robust source of robust enzymes, paving the way for more sustainable and cost-effective bioconversion processes in biofuel and biochemical production and other biotechnological applications. Full article
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<p>Biochemical characterization of xylanase activities in the secretome from the microbiome CMC-50. (<b>A</b>) Temperature optimum; (<b>B</b>) pH optimum; (<b>C</b>) thermostability; (<b>D</b>) storage stability.</p>
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<p>Biochemical characterization of xylanase activities in the secretome from the microbiome XYL-50. (<b>A</b>) Temperature optimum; (<b>B</b>) pH optimum; (<b>C</b>) thermostability; (<b>D</b>) storage stability.</p>
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<p>Native PAGE (<b>A</b>) and zymogram (<b>B</b>) analyses of three independently produced secretomes from the microbiome CMC-50.</p>
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<p>Native PAGE (<b>A</b>) and zymogram (<b>B</b>) analyses of three independently produced secretomes from the microbiome XYL-50.</p>
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<p>Time course of xylan degradation products obtained using the secretome of the consortium XYL-50. Each bar represents the concentration (grams/liter) of xylose (red), xylobiose (green), xylotriose (blue), and xylotetraose (grey) released after 0, 1, 2, 3, 4, 6, and 24 h of digestion.</p>
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<p>Arabinoxylan degradation profiles after digestions using the secretomes of the consortia CMC-50 (<b>A</b>) and XYL-50 (<b>B</b>). Each bar represents the concentration (g/L) of arabinose and xylose released after 0 and 24 h of digestion.</p>
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<p>Phylogenetic tree generated using Classify Microbes with GTDB-Tk. The taxonomic membership of each leaf is represented by a color key corresponding to the outer circle on the plot. The queries are highlighted in yellow, while proximal lineages identified by GTDB-Tk are depicted in lavender.</p>
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<p>Mauve progressive alignment of <span class="html-italic">Bacillus licheniformis</span> CMC-50 and XYL-50.</p>
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<p>Genome comparison among bins extracted from the microbiomes CMC-50 and XYL-50. (Panel <b>A</b>) short-chain fatty acid and alcohol conversions; (Panel <b>B</b>) methanogenesis and methanotrophy; (Panel <b>C</b>) other reductases; (Panel <b>D</b>) sulfur metabolism; and (Panel <b>E</b>) nitrogen metabolism. Numbers at the top indicate (1) <span class="html-italic">Bacillus licheniformis</span> CMC-50; (2) <span class="html-italic">Fictibacillus gelatini</span> XYL-50; (3) <span class="html-italic">Brevibacillus borstelensis</span> XYL-50 and (4) <span class="html-italic">Bacillus licheniformis</span> XYL-50.</p>
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<p>(<b>A</b>) List of operons with CAZymes involved in CMC (Cell operon), xylan/arabinoxylan (Xyl_1, Xyl_2 operons) and arabinan degradation (Arab operon) in the <span class="html-italic">Bacillus licheniformis</span> bins of CMC-50 and XYL-50 microbiomes. (<b>B</b>) Cellulose and xylan/arabinoxylan depolymerization routes proposed from the annotated CAZymes. Predictions of the CAZymes’ mode of action are reported in <a href="#ijms-25-09887-t001" class="html-table">Table 1</a>. “?” indicates that no enzyme predicted to hydrolyze such a bond was identified.</p>
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<p>Simulated xylobiose interaction with GH43_10. MD simulation frame with the higher binding affinity after MD refinement of the GH43_10 AF model docked with xylobiose (<b>A</b>): the model is visualized as a ball-and-stick sketch with interacting residues and xylobiose; carbon atoms of the protein model and xylobiose are differently colored, while oxygen, nitrogen, and polar hydrogens are shown in red, blue, and white; hydrogen bonds are marked as cyan dashed lines; non-covalent interactions of catalytic residues to reactive xylobiose atoms are marked as black dashed lines; a magenta circle is used to indicate the putative position of the water molecule required to be activated for the nucleophilic attack to C<sub>1</sub>, as shown in [<a href="#B40-ijms-25-09887" class="html-bibr">40</a>]. MD simulation statistics (<b>B</b>): different replicates are depicted with different colors. Only frames from 10 to 20 ns were used to generate the distance plots of the atoms involved in the catalysis (marked as black dashed lines in panel (<b>A</b>)); the average and the median values are depicted with a cross and a horizontal line, respectively. C<sub>1</sub>: carbon 1 atom of reducing-end sugar moiety; Ox: oxygen atom.</p>
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13 pages, 622 KiB  
Article
Comparative Study of Different Respiratory Muscle Training Methods: Effects on Cardiopulmonary Indices and Athletic Performance in Elite Short-Track Speedskaters
by Tomasz Kowalski, Andrzej Klusiewicz, Kinga Rębiś, Adrian Wilk and Michał Starczewski
Life 2024, 14(9), 1159; https://doi.org/10.3390/life14091159 - 13 Sep 2024
Viewed by 351
Abstract
Respiratory muscle training (RMT) improves endurance performance, balance, and ability to repeat high-intensity exercise bouts, providing a rationale to be applied in short-track speedskating. To establish a preferable RMT method for short-track speedskating, the influence of inspiratory pressure threshold loading (IPTL) and voluntary [...] Read more.
Respiratory muscle training (RMT) improves endurance performance, balance, and ability to repeat high-intensity exercise bouts, providing a rationale to be applied in short-track speedskating. To establish a preferable RMT method for short-track speedskating, the influence of inspiratory pressure threshold loading (IPTL) and voluntary isocapnic hyperpnoea (VIH) on cardiopulmonary indices and athletic performance was investigated. Sixteen elite short-track speedskaters completed 6 weeks of RMT based on IPTL or VIH. Wingate Anaerobic Tests (WAnTs), cardiopulmonary exercise tests (CPETs), spirometry assessments, and on-ice time trials were performed before and after RMT intervention. Repeated measures ANOVA was used to assess the differences between each method’s influence. No statistically significant (p > 0.05) differences between RMT methods were found in performance during the WAnT, CPET, or specific on-ice time trials. Spirometry measures were similar between both methods. Significant effects were found for the interaction between maximum breathing frequency during CPET (BFmax) and method (p = 0.009), as well as for the interaction between BFMax, method, and sex (p = 0.040). BFmax decreased for IPTL and increased for VIH. The interaction between method and sex revealed that BFmax increased only in males performing VIH. Our findings suggest that IPTL and VIH lead to analogous effects in the study participants, highlighting a negligible practical disparity in the impact of different RMT methods in elite short-track speedskaters. Full article
(This article belongs to the Special Issue Resistance Training Is Medicine)
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<p>Illustrative application of two RMT methods. (<b>A</b>) Presentation of inspiratory pressure threshold loading. (<b>B</b>) Presentation of voluntary isocapnic hyperpnoea.</p>
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14 pages, 1318 KiB  
Article
Anaerobic Digestion of Food Waste with the Addition of Biochar Derived from Microwave Catalytic Pyrolysis of Solid Digestate
by Sofia Lucero Saucedo and Anthony Lau
Sustainability 2024, 16(18), 7997; https://doi.org/10.3390/su16187997 - 13 Sep 2024
Viewed by 626
Abstract
This study explores the potential of biochar derived from microwave-assisted catalytic pyrolysis of solid digestate as an additive to enhance the stability and performance of the anaerobic digestion process. The focus was placed on the effects of biochar dosage, pyrolysis temperature, and pyrolysis [...] Read more.
This study explores the potential of biochar derived from microwave-assisted catalytic pyrolysis of solid digestate as an additive to enhance the stability and performance of the anaerobic digestion process. The focus was placed on the effects of biochar dosage, pyrolysis temperature, and pyrolysis catalyst on methane production. Biochemical methane potential (BMP) tests using synthetic food waste as the substrate revealed a dosage-dependent relationship with specific methane yield (SMY). At a low biochar dosage of 0.1 g/g total solids (TS), improvement in methane (CH4) production was marginal, whereas a high dosage of 0.6 g/g TS increased CH4 content by at least 10% and improved yield by 35–52%. ANOVA analysis indicated that biochar dosage level significantly influenced CH4 yield, while pyrolysis temperature (400 °C vs. 500 °C) and catalyst (20 wt% K3PO4 vs. 10 wt% K3PO4/10 wt% clinoptilolite) did not lead to significant differences in CH4 yield between the treatments. Correlation analysis results suggested that biochar’s most impactful properties on methane yield would be dosage-adjusted specific surface area (or total surface area per unit volume of substrate) and aromaticity index. The findings underscore the potential of solid-digestate-derived biochar as a beneficial additive for anaerobic digestion and hence the sustainability of food waste management system. Full article
(This article belongs to the Section Pollution Prevention, Mitigation and Sustainability)
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<p>Schematic diagram of experimental setup. Created with BioRender.com.</p>
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<p>Methane generation profiles for the experimental treatments: (<b>a</b>) with a low biochar dosage of 0.1 g/g TS; (<b>b</b>) with a high biochar dosage of 0.6 g/g TS as compared to the control.</p>
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<p>Comparison of specific methane yield (SMY) among the experimental treatments.</p>
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10 pages, 1400 KiB  
Article
Analysis of the Laboratory and In-Competition Characteristics of Adolescent Motocross (MX) Riders: An In Situ Case Study
by Ferenc Ihász, Zsolt Katona, Zoltán Alföldi, Attila Szabo, István Barthalos, Robert Podstawski, László Suszter, Kevin J. Finn and László Kerner
Appl. Sci. 2024, 14(18), 8232; https://doi.org/10.3390/app14188232 - 12 Sep 2024
Viewed by 344
Abstract
Motocross is one of the most popular high-speed motorcycle races, which takes place on a naturally closed track with significant challenges. This study aimed to characterize anthropometric, circulatory, and lower and upper limb muscle properties based on laboratory and race-recorded characteristics. Male competitors [...] Read more.
Motocross is one of the most popular high-speed motorcycle races, which takes place on a naturally closed track with significant challenges. This study aimed to characterize anthropometric, circulatory, and lower and upper limb muscle properties based on laboratory and race-recorded characteristics. Male competitors (n = 3) aged 14 years (MX) were included in the study. All three boys have national and international experience. Metabolic characteristics (ventilation, oxygen consumption, and carbon dioxide production) and heart rate (HR) were measured in the laboratory while HR and speed were measured while racing. No significant difference was found between the three competitors in terms of HR during the race. In terms of number of sprints (No. sprint) and maximum speed (Speedmax), we found the most successful runner (highest finisher) to have the most sprints and maximum speed. Comparing the metabolic characteristics, it was found that racing was between the respiratory breakpoint (VT1) and the respiratory compensation point (RCP), but, in several cases, crossed the anaerobic threshold. While motocross riders are exposed to extreme conditions and high physical demands, in motorsport, victories depend not only on the athlete’s physical abilities but also on several factors such as driving technique, mental effort, equipment efficiency and resistance, race strategy, and team competence. Full article
(This article belongs to the Section Applied Biosciences and Bioengineering)
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<p>Description of the racetrack structure: Satellite image of the motocross competition. Legend: Piliscsév, Motorsport Centrum (Hungary). Classification: national and international FIM-registered track. Track length: 1990 m; track width: 6–8 m; level difference: 30 m; number of jumps: 14; number of bends: 13; track surface: earthy clay, sandy in places. Temperature: 18–20 °C; soil quality: ideal, watered. Number of competitors: ~160, in different categories.</p>
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<p>Heart rate (Hr%) patterns of three riders in the first race. Legend: The heart rate pattern of rider (a) is shown by the yellow line, rider (b) is burgundy, and rider (c) is gray.</p>
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<p>Heart rate (Hr%) and speed patterns of one rider. Legend: The red line marked with the number one (1) shows the heart rate pattern, and the blue line with the number two (2) shows the speed; the blue peak with the number three (3) is the speed in the straight section of the track; the blue trough marked with the number four (4) is the reduced speed during the corner; and number five (5) shows the number of completed laps.</p>
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<p>The differences between the heart rate (%) and speed patterns of the three riders tested in the first and second race. Legend: Blue solid line showing rider (a)’s heart rate pattern, orange solid line showing rider (b)’s heart rate pattern, and gray line showing rider (c)’s heart rate pattern. The bottom two graphs show the speed pattern of the same riders.</p>
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13 pages, 4747 KiB  
Article
Effect of Monensin Supplementation in the Bovine Diet on the Composition and Anaerobic Digestion of Manure with and without Screening
by Brenda Kelly Viana Leite, Ana Carolina Amorim Orrico, Marco Antônio Previdelli Orrico Junior, Rusbel Raul Aspilcueta Borquis, Érika Cecília Pereira da Costa, Isabella da Silva Menezes, Juliana Dias de Oliveira and Isabelly Alencar Macena
Fermentation 2024, 10(9), 474; https://doi.org/10.3390/fermentation10090474 - 12 Sep 2024
Viewed by 292
Abstract
The incorporation of monensin into cattle diets can significantly alter the physicochemical properties of excreted manure, potentially affecting waste management and treatment systems given the persistence of substantial concentrations of ionophores in the effluent. This study assessed the impact of monensin on the [...] Read more.
The incorporation of monensin into cattle diets can significantly alter the physicochemical properties of excreted manure, potentially affecting waste management and treatment systems given the persistence of substantial concentrations of ionophores in the effluent. This study assessed the impact of monensin on the compositional characteristics of cattle manure and its implications for anaerobic digestion efficiency, with and without the separation of manure fractions across two hydraulic retention times (HRTs). Manure samples were collected from cattle fed with doses of monensin at 0, 1.8, 3.6, 5.4, and 7.2 mg per kg of dry matter intake. The HRTs investigated were 20 days (HRT20) and 30 days (HRT30). Increasing monensin inclusion in the diets resulted in a notable decrease in the quantities of total solids (TSs), volatile solids (VSs), and neutral detergent fiber (NDF) per animal per day, accompanied by an increase in lignin content and mass. Fraction separation during anaerobic digestion enhances the reduction of TSs, VSs, and NDF, thereby optimizing biogas and methane production potentials and elevating methane concentrations. The presence of monensin correlated with the reduced degradation of organic components during the anaerobic digestion process. To maximize the efficiency of the anaerobic digestion of manure from cattle diets supplemented with monensin, a 30-day HRT combined with fraction separation is recommended. This approach can enhance biogas yield and methane content, thereby improving the sustainability and efficacy of waste treatment processes. Full article
(This article belongs to the Special Issue Biogas and Biochemical Production from Anaerobic Digestion)
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<p>Masses (kg) of (<b>a</b>) total solids (TSs), (<b>b</b>) volatile solids (VSs), (<b>c</b>) neutral detergent fiber (NDF), and (<b>d</b>) lignin produced by cattle fed a diet supplemented with monensin. The shadow area corresponds to the confidence interval (95%).</p>
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<p>(<b>a</b>) Reduction of total solids (TSs, %) and (<b>b</b>) volatile solids (VSs, %) in the anaerobic digestion of waste produced by cattle fed a diet supplemented with monensin and a 20-day hydraulic retention time. SC: screened, US: unscreened. The shadow area corresponds to the confidence interval (95%).</p>
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<p>(<b>a</b>) Reduction of volatile solids (VSs, %) and (<b>b</b>) total solids (TSs, %) during the anaerobic digestion of waste produced by cattle supplemented with doses of monensin and a 30-day hydraulic retention time. SC: screened, US: unscreened. The shadow area corresponds to the confidence interval (95%).</p>
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<p>Reduction of neutral detergent fiber (NDF, %) in the (<b>a</b>) 20-day hydraulic retention time and (<b>b</b>) 30-day hydraulic retention time during the anaerobic digestion of waste produced by cattle supplemented with doses of monensin. SC: screened, US: unscreened. The shadow area corresponds to the confidence interval (95%).</p>
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<p>(<b>a</b>) Potential of biogas production (L/kg of TSs added); (<b>b</b>) methane concentration (%); (<b>c</b>) potential of methane production (L/kg of TSs added) during anaerobic digestion of substrates produced from cattle manure with monensin in the diet and a 20-day hydraulic retention time. SC: screened, US: unscreened. The shadow area corresponds to the confidence interval (95%).</p>
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<p>(<b>a</b>) Potential of biogas production (L/kg of TSs added); (<b>b</b>) methane concentration (%); (<b>c</b>) potential of methane production (L/kg of TSs added) during anaerobic digestion of substrates produced from cattle manure with monensin in the diet and a 30-day hydraulic retention time. SC: screened, US: unscreened. The shadow area corresponds to the confidence interval (95%).</p>
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27 pages, 3426 KiB  
Article
Combination of a Natural Bioactive Compound and High-Hydrostatic-Pressure Treatment to Increase the Shelf-Life and Quality of Pork Meat
by Tamás Csurka, Zoltán Fekete, Anna Visy, Karina Ilona Hidas, István Dalmadi, Ferenc Horváth, László Ferenc Friedrich and Gábor Jónás
Appl. Sci. 2024, 14(18), 8212; https://doi.org/10.3390/app14188212 - 12 Sep 2024
Viewed by 271
Abstract
Currently, there is a growing demand for ready-to-eat, prepared, high-quality, preservative-free products. However, the shelf-life of these products is often so short that a relatively high percentage of these products cannot be sold and end up as food waste. In this study, pork [...] Read more.
Currently, there is a growing demand for ready-to-eat, prepared, high-quality, preservative-free products. However, the shelf-life of these products is often so short that a relatively high percentage of these products cannot be sold and end up as food waste. In this study, pork loin with different marinades (paprika and herbs) was treated with different high hydrostatic pressures (0 MPa, 300 MPa, 450 MPa and 600 MPa) and a bioactive component (piperine) and then the quality of the meat was examined after 0, 4, 8, 12 and 14 days of cold storage. Changes were monitored using color, pH, texture and microbiological analyses. Both pressure, piperine enrichment, storage time and the interactions of different factors had a significant effect on the quality of the loin samples with different marinades. Due to the denaturation of myoglobin, meat slices were less red and lighter after HHP treatment. The addition of piperine reduced this lightness. The pH increased with increasing pressure and decreased with storage time. HHP treatment significantly increased meat hardness, with samples treated at 600 MPa being 19% harder than those treated at 450 MPa. Microbiological results indicated that HHP at 450 MPa and 600 MPa effectively reduced anaerobic total live plate counts, ensuring satisfactory sensory and microbiological quality throughout storage. Piperine fortification also resulted in a more favorable microbiological status during storage without any perceptible change in quality properties during storage. These findings underscore the effectiveness of HHP and piperine enrichment in enhancing the safety and quality of marinated meats. Full article
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<p>Anaerobic total live plate count [lg (CFU g<sup>−1</sup>)] results of paprika-marinated BA− meat sample groups.</p>
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<p>Anaerobic total live plate count [lg (CFU g<sup>−1</sup>)] results of paprika-marinated BA+ meat sample groups.</p>
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<p>Anaerobic total live plate count [lg (CFU g<sup>−1</sup>)] results of herb-marinated BA− meat sample groups.</p>
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<p>Anaerobic total live plate count [lg (CFU g<sup>−1</sup>)] results of herb-marinated BA+ meat sample groups.</p>
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12 pages, 2118 KiB  
Review
Akkermansia muciniphila as a Potential Guardian against Oral Health Diseases: A Narrative Review
by Molly H. Anderson, Karima Ait-Aissa, Amal M. Sahyoun, Ammaar H. Abidi and Modar Kassan
Nutrients 2024, 16(18), 3075; https://doi.org/10.3390/nu16183075 - 12 Sep 2024
Viewed by 473
Abstract
The oral microbiome is a diverse ecosystem containing a community of symbiotic, commensal, and pathogenic microorganisms. One key microorganism linked to periodontal disease (PD) is Porphyromonas gingivalis (P. gingivalis), a Gram-negative anaerobic bacterium known to have several virulence factors that trigger [...] Read more.
The oral microbiome is a diverse ecosystem containing a community of symbiotic, commensal, and pathogenic microorganisms. One key microorganism linked to periodontal disease (PD) is Porphyromonas gingivalis (P. gingivalis), a Gram-negative anaerobic bacterium known to have several virulence factors that trigger inflammation and immune evasion. On the other hand, Akkermansia muciniphila (A. muciniphila), a symbiotic bacterium, has been recently shown to play an important role in mitigating inflammation and reducing periodontal damage. In vivo and in vitro studies have shown that A. muciniphila decreases inflammatory mediators and improves immune responses, suggesting its role in mitigating PD and related inflammatory systemic conditions such as diabetes, hypertension, and obesity. This review discusses the anti-inflammatory effects of A. muciniphila, its impact on periodontal health, and its potential role in managing systemic diseases. The overall aim is to elucidate how this bacterium might help reduce inflammation, improve oral health, and influence broader health outcomes. Full article
(This article belongs to the Section Nutrition and Public Health)
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<p>Oral Bacterial Imbalance, Inflammation, and the Protective Role of <span class="html-italic">Akkermansia muciniphila</span>. Imbalance in oral bacteria leads to inflammation, which contributes to periodontal diseases and can impact systemic health. <span class="html-italic">Akkermansia muciniphila</span>, a gut bacterium with anti-inflammatory properties, can help alleviate these conditions by reducing inflammation and promoting oral and overall health. Red arrows indicate the potentiation or worsening of diseases due to inflammation, while green arrows signify the inhibition or alleviation of diseases by decreasing inflammation.</p>
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<p><span class="html-italic">Akkermansia. muciniphila</span> and inflammation. <span class="html-italic">A. muciniphila</span> by increasing acetate and propionate levels reduces pro-inflammatory cytokines (TNF-α, IL-6, IL-1α, IL-12) while simultaneously increasing the anti-inflammatory cytokine IL-10. These findings highlight the crucial role of <span class="html-italic">A. muciniphila</span> in mitigating inflammation and highlight its potential as a therapeutic agent. Symbol ↑ indicates upregulation/increase and symbol ↓ indicates downregulation/decrease.</p>
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<p><span class="html-italic">A. muciniphila</span> and Oral Health-induced inflammation and bone loss. <span class="html-italic">Porphyromonas gingivalis</span> (<span class="html-italic">P. gingivalis</span>) and <span class="html-italic">Fusobacterium nucleatum</span> (<span class="html-italic">F. nucleatum</span>) have detrimental effects on periodontal diseases (PD) by triggering inflammatory responses and bone destruction. <span class="html-italic">Akkermansia muciniphila</span> has been shown to reduce inflammation, mitigate bone destruction, and enhance anti-inflammatory responses in models of PD. These findings highlight the potential of <span class="html-italic">A. muciniphila</span> as a therapeutic agent in the management of PD-associated inflammation and bone loss. Red arrows indicate the potentiation or worsening of diseases due to inflammation and bone loss, while green arrows signify the inhibition or alleviation of diseases by decreasing inflammation and bone destruction. Symbol ↑ indicates increase.</p>
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<p><span class="html-italic">Akkermansia muciniphila</span> and Other Systemic Diseases Related to Oral Health. <span class="html-italic">Akkermansia muciniphila</span> (<span class="html-italic">A. muciniphila</span>) has emerged as a notable player in managing systemic conditions linked to inflammation, including periodontal disease (PD), obesity, type 2 diabetes (T2D), and hypertension (HTN). <span class="html-italic">A. muciniphila</span>’s influence on inflammatory pathways and metabolic parameters highlights its potential as a therapeutic agent for chronic diseases and emphasizes the importance of the gut microbiome in overall health. Green arrows signify the inhibition or alleviation of diseases by decreasing inflammation.</p>
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<p>Impact of Specific Foods on Metabolic Diseases-induced periodontal diseases and <span class="html-italic">A. muciniphila</span> Abundance. Certain foods that help manage metabolic diseases like hyperlipidemia, diabetes, and hypertension also improve oral health. These foods increase the abundance of <span class="html-italic">Akkermansia muciniphila</span> (<span class="html-italic">A. muciniphila</span>), which may enhance their positive effects on systemic diseases and oral health. This suggests that dietary strategies could effectively address chronic conditions by leveraging <span class="html-italic">A. muciniphila</span>’s role in the gut microbiome to improve overall health.</p>
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12 pages, 1521 KiB  
Article
Physiological Key Determinants of Elite Open-Water Swimmers
by Robin Pla, Yannis Raineteau, Xavier Barbier and Anaël Aubry
Physiologia 2024, 4(3), 305-316; https://doi.org/10.3390/physiologia4030018 - 11 Sep 2024
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Abstract
This study describes the relationships between different physiological measurements and combinations of critical velocity and performance in elite open-water swimmers. A total of 15 international male open-water swimmers performed a 5 × 200 m front crawl incremental test to estimate maximal aerobic speed [...] Read more.
This study describes the relationships between different physiological measurements and combinations of critical velocity and performance in elite open-water swimmers. A total of 15 international male open-water swimmers performed a 5 × 200 m front crawl incremental test to estimate maximal aerobic speed (MAS), maximal oxygen consumption (VO2max), the first lactate threshold (VLT1), the second lactate threshold (VLT2), the speed corresponding to 2 mmol/L (V2mmol/L) and to 4 mmol/L (V4mmol/L), and the lactate threshold (VDmax). A 10 km pool trial was also performed to assess swimming performance. Official competition performances in 200, 400, 800, and 1500 m events were collected and were also used to calculate critical velocity (CV) with various combinations of distances. Personal best performances in 1500 m events were 941.76 ± 20.28 s. For the 10 km trial, performance was strongly correlated to VLT1 and moderately to V2mmol/L (r = 0.73 and 0.67, respectively). For the 400, 800, and 1500 m events, strong correlations were observed for V2mmol/L and V4mmol/L. Moderate correlations with these events were also observed for VLT1 and for VDmax (400 and 800 m only). For long-distance swimmers, assessment with a fixed blood lactate value seems to be a good option to estimate swimming performance in distance events. In addition, 10 km performance seems to be more related to the aerobic threshold than the anaerobic threshold. Full article
(This article belongs to the Special Issue Exercise Physiology and Biochemistry: 2nd Edition)
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<p>Physiological variables from different mathematical methods based on the literature. LT1 = blood lactate value for first lactate threshold, LT2 = blood lactate value for second lactate threshold, D<sub>max</sub> = blood lactate value for D<sub>max</sub> method, V<sub>2mmol/L</sub> = speed corresponding to 2 mmol/L, V<sub>4mmol/L</sub> = speed corresponding to 4 mmol/L, V<sub>LT1</sub> = speed corresponding to first lactate threshold, V<sub>LT2</sub> = speed corresponding to second lactate threshold, and V<sub>Dmax</sub> = speed corresponding to D<sub>max</sub> point.</p>
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<p>Correlation matrix with Pearson’s product moment correlation between physiological capacities and performances. The r values are presented only when <span class="html-italic">p</span>-values were &lt;0.05. Green color indicates positive correlations, while red color indicates negative correlations. V<sub>10km</sub> = speed corresponding to the average velocity during the 10 km pool time trial, SB200 = season best time for 200 m freestyle, SB400 = season best time for 400 m freestyle, SB800 = season best time for 800 m freestyle, SB1500 = season best time for 1500 m freestyle, VO2<sub>max</sub> = maximal oxygen consumption, La<sub>peak</sub> = peak blood lactate concentration at the end of the incremental test, HR<sub>max</sub> = maximal heart rate, MAS = maximal aerobic speed, LT1 = blood lactate value for first lactate threshold, LT2 = blood lactate value for second lactate threshold, Dmax = blood lactate value for Dmax method, V<sub>2mmol/L</sub> = speed corresponding to 2 mmol/L, V4<sub>mmol/L</sub> = speed corresponding to 4 mmol/L, V<sub>LT1</sub> = speed corresponding to first lactate threshold, V<sub>LT2</sub> = speed corresponding to second lactate threshold, and V<sub>Dmax</sub> = speed corresponding to D<sub>max</sub> point.</p>
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<p>Correlation matrix with Pearson’s product moment correlation between critical velocities and performances. The r values are presented only when <span class="html-italic">p</span>-values were &lt;0.05. Green color indicates positive correlations, while red color indicates negative correlations. V<sub>10km</sub> = speed corresponding to the average velocity during the 10 km pool time trial, SB200 = season best time for 200 m freestyle, SB400 = season best time for 400 m freestyle, SB800 = season best time for 800 m freestyle, SB1500 = season best time for 1500 m freestyle, CV200/400 = CV from 200 and 400 m events, CV400/800 = CV from 400 and 800 m events, CV800/1500 = CV from 800 and 1500 m events, CV200/800 = CV from 200 and 800 m events, CV200/1500 = CV from 200 and 1500 events, CV200/400/800 = CV from 200, 400, and 800 m events, CV400/800/1500 = CV from 400, 800, and 1500 m events, and CV200/400/800/1500 = CV from 200, 400, 800, and 1500 m events.</p>
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<p>Correlation matrix with Pearson’s product moment correlation between physiological capacities and critical velocities. The r values are presented only when <span class="html-italic">p</span>-values were &lt;0.05. Green color indicates positive correlations, while red color indicates negative correlations. CV<sub>200/400</sub> = CV from 200 and 400 m events, CV<sub>400/800</sub> = CV from 400 and 800 m events, CV<sub>800/1500</sub> = CV from 800 and 1500 m events, CV<sub>200/800</sub> = CV from 200 and 800 m events, CV<sub>200/1500</sub> = CV from 200 and 1500 events, CV<sub>200/400/800</sub> = CV from 200, 400, and 800 m events, CV<sub>400/800/1500</sub> = CV from 400, 800, and 1500 m events, CV<sub>200/400/800/1500</sub> = CV from 200, 400, 800, and 1500 m events, VO2<sub>max</sub> = maximal oxygen consumption, La<sub>peak</sub> = peak blood lactate concentration at the end of the incremental test, HR<sub>max</sub> = maximal heart rate, MAS = maximal aerobic speed, LT1 = blood lactate value for first lactate threshold, LT2 = blood lactate value for second lactate threshold, Dmax = blood lactate value for Dmax method, V<sub>2mmol/L</sub> = speed corresponding to 2 mmol/L, V4<sub>mmol/L</sub> = speed corresponding to 4 mmol/L, V<sub>LT1</sub> = speed corresponding to first lactate threshold, V<sub>LT2</sub> = speed corresponding to second lactate threshold, and V<sub>Dmax</sub> = speed corresponding to D<sub>max</sub> point.</p>
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20 pages, 1993 KiB  
Article
A New Real-Time Simple Method to Measure the Endogenous Nitrate Reductase Activity (Nar) in Paracoccus denitrificans and Other Denitrifying Bacteria
by José J. García-Trejo, Sharon Rojas-Alcantar, Monserrat Alonso-Vargas, Raquel Ortega, Alejandro Benítez-Guzmán, Leticia Ramírez-Silva, Natalia Pavón, Claudia Peña-Segura, Ofelia Méndez-Romero, Salvador Uribe-Carvajal and Arturo Cadena-Ramírez
Int. J. Mol. Sci. 2024, 25(18), 9770; https://doi.org/10.3390/ijms25189770 - 10 Sep 2024
Viewed by 448
Abstract
The transmembrane nitrate reductase (Nar) is the first enzyme in the dissimilatory alternate anaerobic nitrate respiratory chain in denitrifying bacteria. To date, there has been no real-time method to determine its specific activity embedded in its native membrane; here, we describe such a [...] Read more.
The transmembrane nitrate reductase (Nar) is the first enzyme in the dissimilatory alternate anaerobic nitrate respiratory chain in denitrifying bacteria. To date, there has been no real-time method to determine its specific activity embedded in its native membrane; here, we describe such a new method, which is useful with the inside-out membranes of Paracoccus denitrificans and other denitrifying bacteria. This new method takes advantage of the native coupling of the endogenous NADH dehydrogenase or Complex I with the reduction of nitrate by Nar through the quinone pool of the inner membranes of P. denitrificans. This is achieved under previously reached anaerobic conditions. Inner controls confirming the specific Nar activity determined by this new method were made by the total inhibition of the Nar enzyme by sodium azide and cyanide, well-known Nar inhibitors. The estimation of the Michaelis–Menten affinity of Nar for NO3 using this so-called Nar-JJ assay gave a Km of 70.4 μM, similar to previously determined values. This new Nar-JJ assay is a suitable, low-cost, and reproducible method to determine in real-time the endogenous Nar activity not only in P. denitrificans, but in other denitrifying bacteria such as Brucella canis, and potentially in other entero-pathogenic bacteria. Full article
(This article belongs to the Special Issue Ion Pumps: Molecular Mechanisms, Structure, Physiology)
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<p>Principle of the Nar-JJ coupled assay method. The left side shows the Complex I structure (PDB_ID 4UQ8) assembled in the inner plasma membrane of SBP (sub-bacterial particles), shown as a partial arc (orange transparent bilayer) of a full SBP, simulating a fragment of a pseudo-spherical inverted inner plasma membrane. NADH works as the electron donor that is oxidized by Complex I to NAD<sup>+</sup> (+H<sup>+</sup>, see Equations (1)–(3)), and this reaction is coupled to proton transport across the membrane to the internal aqueous phase of the SBP (dashed arrows). Two electrons from NADH are transferred through the electron transfer prosthetic groups of Complex I to the quinone pool (Q<sub>10</sub>) to produce fully reduced quinol (QH<sub>2</sub>) (see dark thick arrow). The quinone pool works as an electron transfer intermediate between Complex I and the Nar enzyme. Two electrons are then transferred to Nar to reduce NO<sub>3</sub><sup>−</sup> to NO<sub>2</sub><sup>−</sup> with the formation of a water molecule, and scalar protons (see Equations (1)–(3)). This reaction is also coupled to the transfer of protons across the membrane to the inner aqueous space of SBP. No other respiratory complexes participate due to the <span class="html-italic">anaerobic</span> conditions and the inhibition of Complex II or succinate dehydrogenase by malonate. See text for further details.</p>
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<p>The time course of the real-time Nar-JJ assay carried out with SBP from <span class="html-italic">P. denitrificans</span>. The <span class="html-italic">anaerobic</span> reaction cells were totally filled with SBP from <span class="html-italic">P. denitrificans</span> included, and previously depleted of dissolved oxygen by 15 min pre-incubation of succinate respiration. Once oxygen was depleted, 20 mM of malonate was injected to inhibit further Complex II or succinate dehydrogenase activity. This ensures that all electrons used to reduce NO<sub>3</sub><sup>−</sup> by Nar come exclusively from NADH. The nearly zero basal initial slope starting at zero time was read after adding NADH and before the addition of NO<sub>3</sub><sup>−</sup>, and this slow NADH decay is used as basal activity to be subtracted from the coupled NADH/NO<sub>3</sub><sup>−</sup> oxidation-reduction carried out by Complex I and the Nar enzyme, after the addition of NO<sub>3</sub><sup>−</sup> (black thick arrow, bottom). The most linear portion of each trace is used to determine the specific Nar activity of SBP. Red and black traces show duplicates. A representative duplicate of at least 4 different assay determinations is shown.</p>
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<p>The effect of NaN<sub>3</sub> on the Nar-specific activity of SBP from <span class="html-italic">P. denitrificans</span>. The anaerobic reaction cells were prepared as described in <a href="#ijms-25-09770-f002" class="html-fig">Figure 2</a> and in the <a href="#sec3-ijms-25-09770" class="html-sec">Section 3</a>. In the black trace, the basal activity was determined from zero time on the first seconds of reaction before the first 50 s, when the addition of NO<sub>3</sub><sup>−</sup> was carried out as described before. The most linear segment of the negative slope was used to calculate the Nar-specific activity. The red trace shows the Nar activity determined with SBP from <span class="html-italic">P. denitrificans</span> but pre-incubated in the presence of 5 mM of NaN<sub>3</sub> after the addition of 20 mM of malonate. In this case, the nitrate (NO<sub>3</sub><sup>−</sup>) was added before time zero (t ≤ 0) from the very beginning. As shown, the basal activity of the control trace (black) was very similar to the red trace in the presence of 5 mM of NaN<sub>3</sub>. Therefore, the net Nar activity with 5 mM of NaN<sub>3</sub> was essentially zero.</p>
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<p>Determination of the Michaelis–Menten affinity (<span class="html-italic">K<sub>m</sub></span>) of the Nar enzyme for nitrate with the Nar-JJ assay. The CI/Nar coupled reactions were determined at the different concentrations of nitrate ([NO<sub>3</sub><sup>−</sup>]) shown, in <span class="html-italic">anaerobic</span> cells, as described in the <a href="#sec3-ijms-25-09770" class="html-sec">Section 3</a>, at 37 °C. Three independent determinations were averaged, and the standard error (±S.E.) bars are shown for each data plot. The data points were fitted by non-linear regression to the Michaelis–Menten equation in the Origin 7.0 Software and the kinetic values obtained are as follows: <span class="html-italic">V<sub>max</sub></span> = 83 ± 7 nmol/(min.•mg.pt.) and <span class="html-italic">K<sub>m</sub></span> = 70.4 ± 24.7 μM. This value is in the μM range, in concordance with previous determinations of the <span class="html-italic">K<sub>m</sub></span> for NO<sub>3</sub><sup>−</sup>, using natural substrates of the Nar enzyme. See text for further details.</p>
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<p>Determination of the Nar-specific activity in SBP from <span class="html-italic">Brucella canis</span>. The Nar-JJ assays were carried out with SBP prepared from <span class="html-italic">Brucella canis</span> grown under <span class="html-italic">aerobic</span> conditions as described for <span class="html-italic">P. denitrificans</span> in the <a href="#sec3-ijms-25-09770" class="html-sec">Section 3</a>. The red traces show a duplicate of the Nar-JJ assays carried out in <span class="html-italic">anaerobic</span> cells as described in the <a href="#sec3-ijms-25-09770" class="html-sec">Section 3</a>, starting the Nar reaction with 5 mM of NO<sub>3</sub><sup>−</sup> before the zero reading time (t ≤ 0). The black traces show the basal NADH oxidation in the absence of NO<sub>3</sub><sup>−</sup> during the full time course. The quick initial NADH decay in the first 50 s is due to residual oxygen, since these Nar assays were carried out in the absence of glucose oxidase and glucose, which scavenge residual oxygen and abolish these initial decay rates (see for instance <a href="#ijms-25-09770-f004" class="html-fig">Figure 4</a>). The negative slope used to calculate the specific Nar activity of SBP from <span class="html-italic">B. canis</span> (BcSBP) was estimated between the 100 and 300 s of reaction to ensure full linearity of the Nar rate calculations. As can be seen, in <span class="html-italic">B. canis</span>, the Nar-JJ assay gives a reproducible linear NADH decay (red traces) that can be used to calculate the specific BcNar activity after subtraction of the basal NADH decay in the absence of NO<sub>3</sub><sup>−</sup> (black traces). See text for further details.</p>
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<p>Respiratory rates and oxygen depletion in the Nar-JJ assays of “<span class="html-italic">aerobic</span>” and “<span class="html-italic">anaerobic</span>” SBP of <span class="html-italic">P. denitrificans</span>. One Oroboros 2 mL cell was filled with SBP isolated from <span class="html-italic">P. denitrificans</span> cells grown <span class="html-italic">aerobically</span> (red trace) and diluted into the same buffer to the same protein concentration used in the <span class="html-italic">anaerobic</span> spectrophotometric cells of the Nar-JJ assays, i.e., 0.15 mg/mL (see <a href="#sec3-ijms-25-09770" class="html-sec">Section 3</a>). The other Oroboros 2 mL cell was filled identically, but with SBP isolated from <span class="html-italic">P. denitrificans</span> cells grown <span class="html-italic">anaerobically</span> (black trace). The temperature was kept to 37 °C as in the Nar-JJ assays. After about 5 min of basal respiration without addition of respiratory substrates, 10 mM of succinate was injected, and the rate of oxygen consumption was estimated from the most linear segment of each oxygen decay trace. The red trace was repeated 3 times, obtaining a respiratory rate of 944.5 ± 79 natg O/(min.•mg. pt.), and the black trace was repeated 7 times, obtaining 173.1 ± 2.5 natg O/(min.•mg. pt.). The dissolved oxygen was totally consumed in about 3 min, before the 15 min of oxygen depletion by succinate respiration in the case of “<span class="html-italic">aerobic</span>” SBP, and in about 17 min before the 20 min of oxygen depletion used during the Nar-JJ assays with “<span class="html-italic">anaerobic</span>” SBP. In the latter experiments, there was no further addition of glucose oxidase + glucose as in the Nar-JJ assays since the oxygen was totally depleted in both Oroboros cells.</p>
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15 pages, 2827 KiB  
Article
Estimation of Lactate Thresholds, Aerobic Capacity and Recovery Rate from Muscle Oxygen Saturation in Highly Trained Speed Skaters and Healthy Untrained Individuals
by Kinga Rębiś, Andrzej Klusiewicz, Barbara Długołęcka, Paweł Różański, Karol Kowieski and Tomasz Kowalski
J. Clin. Med. 2024, 13(17), 5340; https://doi.org/10.3390/jcm13175340 - 9 Sep 2024
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Abstract
Objective: The main objective of this study was to compare lactate thresholds and aerobic capacity from a graded-intensity exercise test (GXT) for near-infrared spectroscopy measurements in healthy, untrained individuals and highly trained athletes. Methods: This study included 29 untrained students (13 females) and [...] Read more.
Objective: The main objective of this study was to compare lactate thresholds and aerobic capacity from a graded-intensity exercise test (GXT) for near-infrared spectroscopy measurements in healthy, untrained individuals and highly trained athletes. Methods: This study included 29 untrained students (13 females) and 27 highly trained speed skaters (13 females). A maximal effort GXT was performed on a cycloergometer. The lactate-based aerobic and anaerobic thresholds, and the corresponding thresholds for muscle oxygen saturation (SmO2), were determined. Results: The power values determined for all thresholds were significantly higher in female and male speed skaters compared to male and female college students. SmO2 at anaerobic thresholds was significantly lower in female speed skaters than in female students. Both female and male skaters showed greater changes in SmO2 after the GXT compared to students. The recovery did not significantly differ between groups within gender. There was a significant positive correlation in females between the rate of muscle reoxygenation and VO2max power (r = 0.610). In speed skaters, the rate of muscle reoxygenation was not significantly higher than students and correlated positively with VO2max (r = 0.449). Conclusions: The SmO2 at the exercise thresholds, during and after maximal exercise, depends on the training status of the individual. The participants with a higher physical fitness level showed greater decreases in ΔSmO2 at the AT level, as well as after maximal exercise. SmO2 corresponding to the well-established exercise thresholds may be applied to guide training prescription. The rate of muscle reoxygenation after a GXT was also dependent on the aerobic capacity of the participants. Full article
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<p>Location of the MOXY monitor on the vastus lateralis muscle belly.</p>
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<p>Method of evaluating the time for 50% recovery muscle oxygen saturation (SmO<sub>2</sub> recovery half-time, SmO<sub>2</sub>HTR) after the maximal graded exercise test (GXT) for one male student (based on individual data from the present study, compiled according to the guidelines from Nagasawa, 2013).</p>
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<p>Muscle oxygen saturation (SmO<sub>2</sub>) at the aerobic (2 mmol) and anaerobic thresholds (4 mmol, Dmax, LT), expressed as Δ (SmO<sub>2</sub> at Rest − SmO<sub>2</sub> at the aerobic or anaerobic threshold) in the groups of students and speed skaters. *—difference (<span class="html-italic">p</span> &lt; 0.05) between female students and female speed skaters.</p>
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<p>Muscle oxygen saturation (SmO<sub>2</sub>): at baseline, as exercise minimum values, differences between the baseline and minimum (Δ Baseline − Minimum) and the recovery maximum values during the maximal graded exercise test in the groups of students and speed skaters. *—difference (<span class="html-italic">p</span> &lt; 0.05) between female students and female speed skaters, or male students and male speed skaters.</p>
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<p>Time for the 50% recovery of muscle oxygen saturation (SmO<sub>2</sub>) in the groups of students and speed skaters.</p>
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<p>Values of SmO<sub>2</sub> recovery rate in the groups of students and speed skaters. *—difference (<span class="html-italic">p</span> &lt; 0.05) between female students and female speed skaters.</p>
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<p>Statistically significant correlation between the SmO<sub>2</sub> recovery rate and maximal aerobic power (MAP) in the combined groups of female students and speed skaters (n = 26).</p>
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<p>Statistically significant correlation between the SmO<sub>2</sub> recovery rate and VO<sub>2</sub>max in the combined groups of male students and speed skaters (n = 30).</p>
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Article
Exploring the Fermentation Products, Microbiology Communities, and Metabolites of Big-Bale Alfalfa Silage Prepared with/without Molasses and Lactobacillus rhamnosus
by Baiyila Wu, Tong Ren, Changqing Li, Songyan Wu, Xue Cao, Hua Mei, Tiemei Wu, Mei Yong, Manlin Wei and Chao Wang
Agriculture 2024, 14(9), 1560; https://doi.org/10.3390/agriculture14091560 - 9 Sep 2024
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Abstract
The influence of molasses (M) and Lactobacillus rhamnosus (LR) on fermentation products, microbial communities, and metabolites in big-bale alfalfa silage was investigated. Alfalfa (Medicago sativa L.) was harvested at the third growth stage during the flowering stage in the experimental field of [...] Read more.
The influence of molasses (M) and Lactobacillus rhamnosus (LR) on fermentation products, microbial communities, and metabolites in big-bale alfalfa silage was investigated. Alfalfa (Medicago sativa L.) was harvested at the third growth stage during the flowering stage in the experimental field of Linhui Grass Company from Tongliao City, Inner Mongolia. An alfalfa sample without additives was used as a control (C). M (20 g/kg) and LR (106 cfu/g) were added either alone or in combination. Alfalfa was fermented for 7, 14, and 56 d. Lactic acid content in the M, LR, and MLR groups increased, whereas the pH value and butyric acid, 2,3-butanediol, and ethanol contents decreased compared to those of C group after 7, 14, and 56 d of fermentation. A two-way analysis of variance (ANOVA) was performed to estimate the results. The LR group exhibited increased Lactobacillus abundance, whereas the M and MLR groups showed increased Weissella abundance compared to the C group. The relative contents of amino acids (tyrosine, isoleucine, threonine, arginine, valine, and citrulline) in the M and MLR groups were higher than those in the C group. During fermentation, the M, LR, and MLR groups showed decreased phenylalanine, isoleucine, and ferulic acid contents. Amino acids such as isoleucine and L-aspartic acid were positively correlated with Lactobacillus but negatively correlated with Weissella. In conclusion, combining high-throughput sequencing and liquid chromatography–mass spectrometry during anaerobic alfalfa fermentation can reveal new microbial community compositions and metabolite profiles, supporting the application of M, LR, and MLR as feed fermentation agents. Full article
(This article belongs to the Section Farm Animal Production)
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<p>(<b>A</b>) Venn diagram of taxonomic units (OTUs) before and after big-bale alfalfa fermentation. (<b>B</b>) Spatial distribution—principal component analysis of operational taxonomic unit before and after big-bale alfalfa fermentation. FM, fresh material; C, control group; M, molasses treatment group; LR, <span class="html-italic">L. rhamnosus</span> treatment group; MLR, molasses and <span class="html-italic">L. rhamnosus</span> treatment group; Numbers after C, M, LR, and MLR denote fermentation times. The numbers represent OTUs.</p>
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<p>Microbial community at the phylum level (<b>A</b>), genus level (<b>B</b>), and error bar plot (<b>C</b>,<b>D</b>) before and after big-bale alfalfa fermentation. FM, fresh material; C, control group; M, molasses treatment group; LR, <span class="html-italic">L. rhamnosus</span> treatment group; MLR, molasses and <span class="html-italic">L. rhamnosus</span> treatment group; Numbers after C, M, LR, and MLR denote fermentation times.The dot of different colors in the <a href="#agriculture-14-01560-f002" class="html-fig">Figure 2</a>C represent the <span class="html-italic">Lactobacillus</span> abundance. The dot of different colors in the <a href="#agriculture-14-01560-f002" class="html-fig">Figure 2</a>D represent the <span class="html-italic">Weissella</span> abundance.</p>
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<p>Metabolites in big-bale alfalfa silage. (<b>A</b>) Volcano plot analysis of C vs. LR. (<b>B</b>) Volcano plot analysis of C vs. M. (<b>C</b>) Volcano plot analysis of C vs. MLR. C, control group; M, molasses treatment group; LR, <span class="html-italic">L. rhamnosus</span> treatment group; MLR, molasses and <span class="html-italic">L. rhamnosus</span> treatment group.</p>
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<p>Heatmap of differentially accumulated metabolites in big-bale alfalfa silage. C, control group; M, molasses treatment group; LR, <span class="html-italic">L. rhamnosus</span> treatment group; MLR, molasses and <span class="html-italic">L. rhamnosus</span> treatment group; Numbers after C, M, LR, and MLR denote three repetitions.</p>
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<p>Mantel test analyses of microbial communities and metabolites in big-bale alfalfa silage.</p>
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