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17 pages, 2969 KiB  
Article
Newcastle Disease Virus Expressing Cap Gene of Porcine Circovirus Type 2 Confers Protection in Mice and Induced Long-Lasting Neutralizing Antibodies in Pigs
by Sohini Dey, Rudhreswaran Murugasamy, Lukumoni Buragohain, Ajai Lawrence D’silva, Jayashree Sarma, Arpita Bharali, Saravanan Ramakrishnan, Mani Saminathan, Nagendra Nath Barman, Vikram N. Vakharia and Madhan Mohan Chellappa
Vaccines 2024, 12(11), 1285; https://doi.org/10.3390/vaccines12111285 (registering DOI) - 15 Nov 2024
Abstract
Background/Objectives: Porcine Circovirus 2 (PCV2) infection poses significant health and economic challenges to the global swine industry. The disease in pigs leads to lymphoid depletion, resulting in immunosuppression and increased susceptibility to co-infections with other bacterial and viral pathogens. This study evaluated [...] Read more.
Background/Objectives: Porcine Circovirus 2 (PCV2) infection poses significant health and economic challenges to the global swine industry. The disease in pigs leads to lymphoid depletion, resulting in immunosuppression and increased susceptibility to co-infections with other bacterial and viral pathogens. This study evaluated the efficacy of two novel recombinant Newcastle disease virus (NDV) strain R2B vectored vaccines that express the cap gene of PCV2 alone and along with the transmembrane and cytoplasmic tail (TMCT) domains of the NDV F gene. The efficacy of the vaccine candidates was studied in mouse and pig models. Methods: Six-week-old BALB/c mice were divided into five groups and immunized intramuscularly three times at 14-day intervals with various vaccine candidates, namely rNDV-R2B-PCVcap-TMCT, rNDV-R2B-PCVcap, and CircoFLEX commercial vaccine, along with controls. Following immunization and PCV2d virus challenge, multiple assays assessed the immune responses in animal trials. In the pig animal trial, pigs were divided into four groups: a control group (PBS), NDV-vectored PCVcap-TMCT group, NDV-vectored-PCVcap group, and CircoFLEX vaccine group. Pigs were immunized intramuscularly twice at 28-day intervals. Blood samples were collected at regular intervals over 70 days to evaluate the humoral and cell-mediated immune responses. Results: Both mice and pigs’ trials indicated that the NDV-vectored PCV2 cap-TMCT vaccine candidate elicited superior immune responses. In mice, the rNDV-R2B-PCVcap-TMCT group showed enhanced humoral and cellular immunity, increased PCV2-specific antibody levels, higher CD4+/CD8+ ratio, elevated IFN-γ and TNF-α levels, decreased IL-10 levels, reduced viral loads, and minimal histopathological changes. In pigs, the NDV-vectored PCVcap-TMCT group demonstrated better antibody responses, cytokine profiles (IFN-γ and IL-10), and higher levels of PCV2-specific neutralizing antibodies against the PCV2a, PCV2b and PCV2d genotypes when compared to other groups. Conclusions: These findings suggest NDV-vectored PCVcap-TMCT vaccine candidate, expressing the cap gene of PCV2 along with the TMCT domain, offers a promising alternative for protecting against PCV2 infection, potentially addressing the challenges posed by emerging PCV2 strains in the swine industry. Full article
(This article belongs to the Special Issue Strategies of Viral Vectors for Vaccine Development)
12 pages, 7492 KiB  
Article
Porcine Airway Organoid-Derived Well-Differentiated Epithelial Cultures as a Tool for the Characterization of Swine Influenza a Virus Strains
by Nora M. Gerhards, Manouk Vrieling, Romy Dresken, Sophie Nguyen-van Oort, Luca Bordes, Jerry M. Wells and Rik L. de Swart
Viruses 2024, 16(11), 1777; https://doi.org/10.3390/v16111777 - 15 Nov 2024
Viewed by 87
Abstract
Swine influenza A viruses (IAVsw) are important causes of disease in pigs but also constitute a public health risk. IAVsw strains show remarkable differences in pathogenicity. We aimed to generate airway organoids from the porcine lower respiratory tract and use these to establish [...] Read more.
Swine influenza A viruses (IAVsw) are important causes of disease in pigs but also constitute a public health risk. IAVsw strains show remarkable differences in pathogenicity. We aimed to generate airway organoids from the porcine lower respiratory tract and use these to establish well-differentiated airway epithelial cell (WD-AEC) cultures grown at an air–liquid interface (ALI) for in vitro screening of IAVsw strain virulence. Epithelial cells were isolated from bronchus tissue of juvenile pigs, and airway organoids were cultured in an extracellular matrix in a culture medium containing human growth factors. Single-cell suspensions of these 3D organoids were seeded on Transwell filters and differentiated at ALI to form a pseudostratified epithelium containing ciliated cells, mucus-producing cells and tight junctions. Inoculation with a low dose of IAVsw in a low volume inoculum resulted in virus replication without requiring the addition of trypsin, and was quantified by the detection of viral genome loads in apical washes. Interestingly, inoculation of an H3N2 strain known to cause severe disease in pigs induced a greater reduction in trans-epithelial resistance and more damage to tight junctions than H1N2 or H1N1 strains associated with mild disease in pigs. We conclude that the porcine WD-AEC model is useful in assessing the virulence of IAVsw strains. Full article
(This article belongs to the Special Issue Endemic and Emerging Swine Viruses 2024)
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Figure 1

Figure 1
<p>Establishment of AO-derived WD-AECs. (<b>A</b>) Schematic outline of the isolation of porcine bronchial epithelial cells from the primary bronchi which were subsequently grown as 3D organoids in an extracellular matrix before seeding in 2D on Transwell filters and culturing at air–liquid interface upon confluency. (<b>B</b>) Transverse histology sections over the course of 7 weeks showing the development of a pseudostratified ciliated respiratory epithelium. Hematoxylin and eosin stain, 40× objective. (<b>C</b>) Immunohistochemistry (IHC) staining of WD-AECs 7 weeks post-airlift and porcine bronchus epithelial cells. P63 staining to visualize basal cells, Muc5AC staining to visualize mucus (goblet cells) and acetylated tubulin staining to visualize cilia. WD-AEC resembles an <span class="html-italic">in vivo</span> bronchial epithelium in cellular composition and morphology, despite a reduced thickness. (<b>D</b>) Development of transepithelial electrical resistance (TEER) over the course of differentiation of WD-AECs. After an initial increase post-airlift, TEER values remained consistent. For B, C and D: Differentiation was followed for three separate donors. Representative data from one experiment are shown.</p>
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<p>Development of WD-AECs over time. Confocal laser scanning microscopy images showing the development of tight junctions (ZO-1), goblet cells (Muc5ac) and cilia (acetylated tubulin) over the course of 6 weeks after airlifting Transwell filters. Goblet cells were most abundant at 2–4 weeks, while cilia appeared after 3 weeks. Tight junctions were detected from week 0 onwards, but only started to show a regular pattern from week 2 onwards. Merged images show nuclei in blue, tight junctions in red, goblet cells in green and cilia in yellow. Scale bar represents 100 µm. 40× objective. Representative data from one experiment are shown.</p>
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<p>Infection of porcine WD-AECs with IAVsw. (<b>A</b>) Experimental design of IAVsw inoculation of porcine WD-AEC cultures. Three different IAVsw strains were used (H1N1, H1N2, H3N2) and applied apically at a dose of 10<sup>2</sup> or 10<sup>3</sup> TCID<sub>50</sub> in a volume of 10 µL. After 1 h incubation, Transwell filters were washed 4 times. An apical wash was collected at 16 h, 24 h and 48 h post-inoculation, and transepithelial electrical resistance (TEER) was measured at 0 hpi, 24 hpi and 48 hpi. At 48 hpi, filters were fixed and stained by hematoxylin and eosin (HE) and against IAV nucleoprotein (NP). (<b>B</b>) Viral RNA loads in apical washes. All viruses replicated in porcine WD-AECs. For H1N2, a dose–response curve was observed over all evaluated time points, which was only observed at the earlier time points for the other two viruses. (<b>C</b>) TEER values as a measure of epithelial integrity. TEER values declined for H3N2- and H1N1-inoculated WD-AECs over 48 h but remained constant in H1N2-inoculated WD-AECs. (<b>D</b>) Transverse histology sections stained by HE at 48 h post-infection. The epithelial cell layer remained intact for H1N2-inoculated WD-AECs, while there was substantial thinning of the cell layer in H3N2-inoculated WD-AECs. For H1N1, the higher dose seemed to cause more cell loss compared to the lower dose. 40× objective. B, C and D show representative results from one out of three experiments (pig04).</p>
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<p>IAV NP expression after 24 h post-infection with 10<sup>3</sup> TCID<sub>50</sub><b>.</b> Confocal microscopy images to visualize expression of IAV NP and epithelial composition. (<b>A</b>) H1N1-inoculated WD-AECs showed NP expression (green), continuous tight junctions (red), a thick layer of cilia (yellow) and predominantly intact nuclei (blue). (<b>B</b>) H3N2-inoculated filters showed NP expression while at the same time tight junctions were compromised, cilia expression was reduced and more fragmented nuclei were visible. (<b>C</b>) Angled side-view of WD-AEC shown in (<b>A</b>). (<b>D</b>) Angled side-view of WD-AEC shown in (<b>B</b>). Note that IAV NP staining (green) is visible within or above the apical side of WD-AEC cultures. All images: Scale bar represents 50 µm. 100× objective. Representative data from one experiment are shown.</p>
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19 pages, 7052 KiB  
Article
Insights into the Acute Stress of Glutaraldehyde Disinfectant on Short-Term Wet Anaerobic Digestion System of Pig Manure: Dose Response, Performance Variation, and Microbial Community Structure
by Yongming Wu, Fangfei Li, Liuxing Wu, Shifu He, Peiyu Liang, Lei Zhang, Zhijian Wu, Tao Zhang, Yajun Liu, Xiangmin Liu, Xueping Huang, Lin Zhu, Maolin Wang and Mi Deng
Water 2024, 16(22), 3279; https://doi.org/10.3390/w16223279 - 15 Nov 2024
Viewed by 165
Abstract
The outbreak of epidemics such as African swine fever has intensified the use of disinfectants in pig farms, resulting in an increasing residual concentration of disinfectants in environmental media; however, the high-frequency excessive use of disinfectants that damage pig farm manure anaerobic fermentation [...] Read more.
The outbreak of epidemics such as African swine fever has intensified the use of disinfectants in pig farms, resulting in an increasing residual concentration of disinfectants in environmental media; however, the high-frequency excessive use of disinfectants that damage pig farm manure anaerobic fermentation systems and their mechanisms has not attracted enough attention. Especially, the complex effects of residual disinfectants on anaerobic fermentation systems for pig manure remain poorly understood, thus impeding the application of disinfectants in practical anaerobic fermentation systems. Herein, we explored the effects of glutaraldehyde disinfectant on methane production, effluent physicochemical indices, and microbial communities in a fully automated methanogenic potential test system (AMPTSII). The results show that adding glutaraldehyde led to remarkable alterations in methane production, chemical oxygen demand (COD), volatile solids (VS), and polysaccharide and phosphorus concentrations. During the anaerobic process, the production of methane displayed a notable decrease of 5.0–98% in all glutaraldehyde treatments, and the trend was especially apparent for treatments containing high levels of glutaraldehyde. Comparisons of the effluent quality showed that in the presence of 0.002–0.04% glutaraldehyde, the COD and total phosphorus (TP) increased by 12–310% and 15–27%, respectively. Moreover, the addition of 0.01–0.08% glutaraldehyde decreased the ammonium (NH4+-N) concentration and VS degradation rate by 7.7–15% and 4.9–26.2%. Furthermore, microbiological analysis showed that the glutaraldehyde treatments had adverse effects on the microbial community. Notably, certain functional bacteria were restrained, as highlighted by the decreases in relative abundance and microbial diversity by 1.3–17% and 0.06–21%, respectively. This study provides a theoretical basis for the rational use of disinfectants in anaerobic fermentation systems. Full article
(This article belongs to the Special Issue The Control of Legacy and Emerging Pollutants in Soil and Water)
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Figure 1
<p>Fully automated methane potential test system (AMPTS II): (<b>a</b>) Anaerobic fermentation unit, (<b>b</b>) CO<sub>2</sub> adsorption unit, (<b>c</b>) Computing and data processing.</p>
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<p>Effects of glutaraldehyde on effluent properties during the anaerobic fermentation process: (<b>a</b>) Chemical oxygen demand (COD), (<b>b</b>) Total phosphorus (TP), (<b>c</b>) Ammonia nitrogen (NH<sub>4</sub><sup>+</sup>-N), (<b>d</b>) Total nitrogen (TN), (<b>e</b>) Polysaccharide (NH<sub>4</sub><sup>+</sup>-N), (<b>f</b>) pH value, (<b>g</b>) Volatile suspended solids (VS), (<b>h</b>) Total suspended solids (TS). *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>(<b>a</b>,<b>b</b>) Effects of glutaraldehyde on methane production in anaerobic fermentation systems. *, <span class="html-italic">p</span> &lt; 0.05; ***, <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effect of glutaraldehyde on microbial Alpha diversity index and dysbiosis index in anaerobic fermentation systems: (<b>a</b>) Chao Indices, (<b>b</b>) Ace Indices, (<b>c</b>) Shannon Indices, (<b>d</b>) Simpson Indices, (<b>e</b>) MDI index of pre-reaction, (<b>f</b>) MDI index of post-reaction. *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effects of glutaraldehyde on the microbial community structure of anaerobic sludge: (<b>a</b>) Phylum level, (<b>b</b>) genus level, (<b>c</b>) PCoA analysis, (<b>d</b>) Chloroflexi Pre-reaction changes, (<b>e</b>) <span class="html-italic">Syntrophomonas</span> Pre-reaction changes, and (<b>f</b>) <span class="html-italic">Bacteroides</span> Post-reaction changes. *, <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Correlations among main microbial species and key physicochemical indices: (<b>a</b>) redundancy analysis (RDA) and (<b>b</b>) Correlation of environmental factors. *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Functional and community analysis of related genes: (<b>A</b>,<b>B</b>) Gene functions of microflora at different stages, (<b>C</b>) Venn diagram for pre-reaction genus levels, (<b>D</b>) Venn diagram for post-reaction genus levels.</p>
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18 pages, 2090 KiB  
Article
From Structure to Strength: Analyzing the Impact of Sulfuric Acid on Pig Bone Demineralization Through FTIR, LIBS, and AAS
by Milica Marković, Miroslav Kuzmanović, Dragan Ranković, Danica Bajuk-Bogdanović, Aleksandra Šajić and Dušan Dimić
Int. J. Mol. Sci. 2024, 25(22), 12250; https://doi.org/10.3390/ijms252212250 - 14 Nov 2024
Viewed by 224
Abstract
The present research aimed to investigate the demineralizing effects of sulfuric acid on pig bone. Alterations in collagen and phosphate contents and changes in the elemental composition of the bone during the 14-day-long immersion in sulfuric acid solutions of different concentrations were estimated [...] Read more.
The present research aimed to investigate the demineralizing effects of sulfuric acid on pig bone. Alterations in collagen and phosphate contents and changes in the elemental composition of the bone during the 14-day-long immersion in sulfuric acid solutions of different concentrations were estimated using ATR-FTIR, LIBS, and AAS. FTIR spectra at amide I (1800–1600 cm−1) and phosphate ν13 (PO43−) (1300–900 cm−1) domains were scrutinized using the deconvolution method for monitoring changes in the protein secondary structure and mineral content. The results implicated sulfuric acid as a powerful demineralization agent and effective in targeting mineral components, such as hydroxyapatite, while leaving the collagen matrix relatively preserved with a complex secondary structure. Collagen maturity marker values gave valuable insights into the structural integrity of the bone. LIBS and AAS indicated changes in bone hardness; phosphorous-to-carbon ratio; and calcium, phosphorous, and magnesium content in the solutions left after the immersion period. The changes in the ratio of ionic-to-atomic calcium lines in the LIBS spectra indicated hardening of the bone, with increasing acid concentration and prolonged action, due to the deposition of calcium sulfate on the surface. The calcium concentration in the solutions decreased with increased acid concentration, while the change in phosphorus and magnesium concentrations was reversed. Full article
(This article belongs to the Special Issue Molecular Updates and Applications in Forensic Medicine)
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Graphical abstract

Graphical abstract
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<p>Pig shoulder bone after immersion in sulfuric acid.</p>
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<p>Comparative display of ATR–FTIR spectra of pig shoulder bone in sulfuric acid (0.01, 0.1, and 1.0 M) after immersion periods of 1 (<b>a</b>), 7 (<b>b</b>), and 14 (<b>c</b>) days.</p>
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<p>Comparative display of ATR–FTIR spectra of pig shoulder bone in sulfuric acid (0.01, 0.1, and 1.0 M) after immersion periods of 1 (<b>a</b>), 7 (<b>b</b>), and 14 (<b>c</b>) days.</p>
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<p>Collagen maturity marker values are calculated as the area ratio of the sub-bands at 1660 and 1690 cm<sup>−1</sup>.</p>
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<p>The results of the curve fitting analysis in the 1200–900 cm<sup>−1</sup> IR region of pig bone in 1.0 M sulfuric acid: immersion time, 1 (<b>a</b>), 7 (<b>b</b>), and 14 (<b>c</b>) days.</p>
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<p>LIBS spectra of pig shoulder bone: Ca II and Ca I lines (0.1 M, different days) (<b>a</b>) and P I and C I lines ((<b>b</b>), day 1, different concentrations).</p>
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<p>Intensity ratio of Ca II/Ca I lines (364.441/370.603 nm) (<b>a</b>) and P/C I lines (247.86/255.33 nm) (<b>b</b>) obtained from LIBS spectra.</p>
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<p>Concentration of calcium (<b>a</b>), phosphorous (<b>b</b>), and magnesium (<b>c</b>) in remaining solutions obtained by AAS.</p>
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13 pages, 7485 KiB  
Article
A Comparative Metabolomics Study of the Potential Marker Compounds in Feces from Different Hybrid Offspring of Huainan Pigs
by Yufu Li, Mingyang Jia, Junfeng Chen, Fujiu Liu, Qiaoling Ren, Xiangzhou Yan, Baosong Xing, Chuanying Pan and Jing Wang
Animals 2024, 14(22), 3282; https://doi.org/10.3390/ani14223282 - 14 Nov 2024
Viewed by 191
Abstract
As a notable native Chinese genetic population, the Huainan pig has an exceptional meat quality but a low percentage of lean meat and subpar genetic performance. To better exploit the superior genetic traits of the Huainan pig and address knowledge gaps regarding the [...] Read more.
As a notable native Chinese genetic population, the Huainan pig has an exceptional meat quality but a low percentage of lean meat and subpar genetic performance. To better exploit the superior genetic traits of the Huainan pig and address knowledge gaps regarding the optimization of its hybrid offspring, this study used Huainan pigs as the maternal line and bred them with Yorkshire, Landrace, and Berkshire sires. This approach produced three hybrid combinations: Yorkshire × Huainan (YH), Landrace × Huainan (LH), and Berkshire × Huainan (BH). The body size, fat ratio, and average backfat thickness of these hybrid progeny were evaluated under the same feeding management and nutritional circumstances. The results revealed that the average backfat thickness of YH was significantly lower than that of LH and BH. In order to better understand the causes of these variations, fecal samples were taken from three pigs in each group for metabolomic analysis. A total of 2291 metabolites were identified, including benzene derivatives (16.6%), amino acids and their metabolites (14.5%), and organic acids (13.4%), with pyruvaldehyde and norethindrone acetate elevated in YH compared to LH and BH. In addition, the three hybrid pig groups commonly exhibited differences in the “glycerophospholipid metabolism” pathway. This variation may also contribute to differences in their fat ratio and backfat thickness. Our findings provide a novel perspective on the role of hybrid vigor in advancing the genetic population of Huainan pigs, while also revealing the unique metabolic characteristics of the YH with regard to fat deposition. This study is expected to enhance the conservation and effective utilization of genetic resources within the Huainan pig population. Full article
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<p>The overall workflow of the hybrid experiment and metabolomics strategy.</p>
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<p>Comparative analysis of the three hybrid pig combinations in terms of weight (<b>A</b>), body height (<b>B</b>), body length (<b>C</b>) chest girth (<b>D</b>) fat ratio (<b>E</b>), and average backfat thickness (<b>F</b>). “*” means: <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Composition (<b>A</b>) and clustering analysis heatmap (<b>B</b>) of metabolites in feces from three hybrid pig combinations. Colors correspond to the distinct values achieved following relative content normalization (red denotes high levels and green denotes low levels).</p>
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<p>Investigation of DAMs across the three distinct hybrid pig combinations. Fecal metabolite profiling was performed using OPLS-DA models between (<b>A</b>) YH and LH, (<b>B</b>) YH and BH, and (<b>C</b>) LH and BH participants. Volcano graphs (<b>D</b>–<b>F</b>) showing the DAMs for the three groups. (<b>G</b>) Upset plots showing the overlapping and accession-specific DAMs.</p>
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<p>The KEGG enrichment plots show the metabolic pathways enriched with specific metabolites that are expressed differently between YH and LH (<b>A</b>), YH and BH (<b>B</b>), and LH and BH (<b>C</b>). The <span class="html-italic">x</span>-axis signifies the Rich Factor associated with each pathway, while the <span class="html-italic">y</span>-axis shows the names of the pathways arranged in order of their <span class="html-italic">p</span>-value. The color of the data points reflects the size of the <span class="html-italic">p</span>-value, where red shades suggest a higher level of enrichment. The magnitude of the data points corresponds to the quantity of metabolites that are differentially expressed and enriched in that particular pathway. (<b>D</b>) Upset plots showing the overlapping pathways.</p>
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23 pages, 2473 KiB  
Article
Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs
by Ester Sevillano, Irene Lafuente, Nuria Peña, Luis M. Cintas, Estefanía Muñoz-Atienza, Pablo E. Hernández and Juan Borrero
Int. J. Mol. Sci. 2024, 25(22), 12210; https://doi.org/10.3390/ijms252212210 - 14 Nov 2024
Viewed by 261
Abstract
Antimicrobial resistance (AMR) poses a significant challenge to animal production due to the widespread use of antibiotics. Therefore, there is an urgent need for alternative antimicrobial strategies to effectively manage bacterial infections, protect animal health, and reduce reliance on antibiotics. This study evaluated [...] Read more.
Antimicrobial resistance (AMR) poses a significant challenge to animal production due to the widespread use of antibiotics. Therefore, there is an urgent need for alternative antimicrobial strategies to effectively manage bacterial infections, protect animal health, and reduce reliance on antibiotics. This study evaluated the use of emerging approaches and procedures for the isolation, identification, and characterization of bacteriocin-producing bacteria and their bacteriocins, sourced from the gastrointestinal tract (GIT) of meat-producing pigs. Out of 2056 isolates screened against Gram-positive and Gram-negative indicator strains, 20 of the most active antimicrobial isolates were subjected to whole genome sequencing (WGS) for the prediction of coding DNA sequences (CDS) and the identification of bacteriocin gene clusters (BGC) and their functions. The use of an in vitro cell-free protein synthesis (IV-CFPS) protocol and the design of an IV-CFPS coupled to a split-intein mediated ligation (IV-CFPS/SIML) procedure made possible the evaluation of the production and antimicrobial activity of described and putatively novel bacteriocins. A colony MALDI-TOF MS procedure assisted in the identification of class I, II, and III lanthipeptides. MALDI-TOF MS and a targeted proteomics, combined with a massive peptide analysis (LC-MS/MS) approach, has proven valuable for the identification and biochemical characterization of previously described and novel bacteriocins encoded by the isolated bacteriocin-producing strains. Full article
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<p>Alignment of the bacteriocin gene clusters (BGC) from <span class="html-italic">E. coli</span> PG14 and <span class="html-italic">E. coli</span> PG15 isolates, which encode identical bacteriocins as performed using BAGELv.4.0. ORFs are represented by arrows and those with a predicted function are indicated by gene identity and/or color. Red squares highlight regions where gene rearrangements have been identified.</p>
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<p>(<b>a</b>). Bacteriocin gene cluster (BGC) and antimicrobial activity assessed by the spot-on-agar test (SOAT) of IV-CFPS-produced class II bacteriocins from <span class="html-italic">L. salivarius</span> P1CEA3, <span class="html-italic">L. salivarius</span> PG21, and <span class="html-italic">S. saprophyticus</span> P1CEA4 against different indicator strains. (<b>b</b>). BGC and antimicrobial activity evaluated by the spot-on-agar test (SOAT) of IV-CFPS/SIML-produced putative class I circular bacteriocins PL1 and PL2, encoded by <span class="html-italic">P. lentus</span> P8CEA5, against different indicator strains. ORFs are represented by arrows and those with a predicted function are indicated by gene identity and/or color.</p>
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<p>Colony MALDI-TOF MS analysis of samples from <span class="html-italic">L. salivarius</span> P1CEA3, <span class="html-italic">P. dendritiformis</span> P1CEA1, <span class="html-italic">P. lentus</span> P8CEA5, and <span class="html-italic">S. simulans</span> P8CEA5, which encode different class I, II, and III lanthipeptide bacteriocins. The BGC and amino acid sequences of the lanthipeptides are represented. ORFs are represented by arrows and those with a predicted function are indicated by gene identity and/or color.</p>
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<p>MALDI-TOF MS analysis of the RP-FPLC active fractions from the purified supernatants of <span class="html-italic">P. lentus</span> P8CEA5. (<b>a</b>). Sinapinic acid matrix, and (<b>b</b>). α-cyano-4-hydroxycinnamic acid matrix. Peptide peaks with the molecular masses as observed for the predicted lentucin S (thuricin 17 family bacteriocin), thermophilin A LP (TheA) and the circular bacteriocins PL1 and PL2.</p>
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11 pages, 803 KiB  
Article
Dry-Cured Ham, ‘Kraški Pršut’, from Heavy Pig Production—A Pilot Study Focusing on the Effect of Ham Weight and Salting
by Bojana Savić, Marjeta Čandek-Potokar and Martin Škrlep
Foods 2024, 13(22), 3620; https://doi.org/10.3390/foods13223620 - 13 Nov 2024
Viewed by 334
Abstract
A pilot study was conducted with the aim of adapting the processing of “Kraški pršut”, dry-cured ham, for thighs from heavy pigs. The focus was on the effect of ham weight and salting duration on the quality of dry-cured ham. From [...] Read more.
A pilot study was conducted with the aim of adapting the processing of “Kraški pršut”, dry-cured ham, for thighs from heavy pigs. The focus was on the effect of ham weight and salting duration on the quality of dry-cured ham. From a pool of thighs harvested from heavy pigs, a total of 32 green hams were selected (from 16 carcasses) based on weight (two classes; L—lighter, H—heavier) and we used left and right ham for either the standard or a shortened salting phase. Salting duration consisted of phase 1 (7 days for all hams) and phase 2 (7 or 14 days for L, 10 or 17 days for H, in the case of shortened and standard salting, respectively). Equivalent conditions for all hams were maintained during the remaining phases, with a total maturation period of 18 months. The analysis focused on chemical, physical and rheological properties, sensory attributes, and consumer perceptions. The H hams had lower processing losses, resulting in higher moisture and water activity, lower salt content in internal biceps femoris muscle, and a softer texture (instrumental and sensory) than L hams. The salting duration mainly affected weight losses in the salting phase and, consequently, salt content, which was lower in the shortened salting phase, while no effects were observed on texture. The sensory panel perceived weight’s effect on hardness, with L hams being perceived as harder, and salting’s effect on sourness, with hams submitted to longer salting perceived as sourer than H hams. Consumer testing indicated a general preference for softer and less salty hams. Overall, the results show that the applied reduction in salting duration was not substantial, and future trials should explore further optimization in terms of salting and resting phases. Full article
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<p>Experimental design.</p>
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<p>Consumer perceptions as affected by ham weight and salting duration—figures present the frequency (%) of perceived descriptors using the check all that apply (CATA) method. Significance: * = <span class="html-italic">p</span> &lt; 0.05; ** = <span class="html-italic">p</span> &lt; 0.01; *** = <span class="html-italic">p</span> &lt; 0.001.</p>
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18 pages, 2447 KiB  
Article
Effects of Acorns on Fatty Acid Composition and Lipid Metabolism in Adipose Tissue of Yuxi Black Pigs
by Zhe Sun, Dongyang Liu, Siyuan An, Xuejia Wu, Jinzhou Zhang and Zhiguo Miao
Animals 2024, 14(22), 3271; https://doi.org/10.3390/ani14223271 - 13 Nov 2024
Viewed by 246
Abstract
The objective of the present research was the examination of how varying proportions of an acorn diet affects the deposition of subcutaneous fat and the composition of fatty acids (FAs) in Yuxi black pigs. Ninety pigs (with a balanced sex ratio and a [...] Read more.
The objective of the present research was the examination of how varying proportions of an acorn diet affects the deposition of subcutaneous fat and the composition of fatty acids (FAs) in Yuxi black pigs. Ninety pigs (with a balanced sex ratio and a similar weight 99.60 ± 2.32 kg) were stochastically assigned to the control group (CON) and the dietary acorn experimental groups (AEG). The CON was fed basal diets and the AEG1, AEG2, AEG3, and AEG4 groups were provided with dietary regimens comprising twenty, thirty, forty, and fifty per cent acorns, respectively. Each group consisted of six pigs, with three replicates. The breeding cycle was four months. The results demonstrated that, in comparison with the CON group, the lean meat rate was significantly increased in all test groups (p < 0.05), while in backfat thickness, loin eye area, carcass weight and slaughter rate was no significant difference (p > 0.05). The serum TC/HDL (total cholesterol divided by high-density lipoprotein-cholesterol) and TG/HDL (Triglyceride divided by high-density lipoprotein-cholesterol) levels in the AEG1 and AEG2 groups were significantly lower than the CON group (p < 0.05). There was no significant effect on the composition of FAs (p > 0.05). The number of fat cells in subcutaneous back fat and subcutaneous abdominal fat was significantly increased, and the area of fat cells was decreased (p < 0.05). Furthermore, the levels of ATGL and HSL expression in the subcutaneous back fat, as well as ACC, FAS, ATGL, PPARγ, and HSL expression in the subcutaneous abdominal fat, were significantly increased in the AEG2 group compared to the CON group (p < 0.05). Additionally, the expression of ACC, FAS, FABP4, PPARγ, C/EBPα, and FAS/HSL in the subcutaneous back fat, as well as FABP4, C/EBPα, and FAS/HSL in the subcutaneous abdominal fat, were significantly lower in the AEG2 group compared to the CON group (p < 0.05). In conclusion, it has been found that a 30% acorn diet can inhibit subcutaneous fat deposition and enhance the nutritional value of pork and the health of Yuxi black pigs. Full article
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<p>Subcutaneous adipose tissue morphology HE (Hematoxylin-Eosin staining) × 20.</p>
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<p>Lipid metabolism genes expression in subcutaneous back fat. The vertical bar represents the standard errors. The presence of lower-case letters indicate a statistically significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Lipid metabolism genes expression in subcutaneous abdominal fat. The vertical bar represents the standard errors. The presence of lower-case letters indicate a statistically significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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18 pages, 1689 KiB  
Article
Effects of Flavonoids in Fructus Aurantii Immaturus on Carcass Traits, Meat Quality and Antioxidant Capacity in Finishing Pigs
by Zekun Yang, Qiuping Guo, Xiangfeng Kong, Yixing Li and Fengna Li
Antioxidants 2024, 13(11), 1385; https://doi.org/10.3390/antiox13111385 - 13 Nov 2024
Viewed by 426
Abstract
This experiment aimed to explore the effects of flavonoids in Fructus Aurantii Immaturus (FFAI) on carcass traits, meat quality, and the antioxidant capacity of finishing pigs. The results indicated that the addition of an appropriate amount of FFAI into their diet could significantly [...] Read more.
This experiment aimed to explore the effects of flavonoids in Fructus Aurantii Immaturus (FFAI) on carcass traits, meat quality, and the antioxidant capacity of finishing pigs. The results indicated that the addition of an appropriate amount of FFAI into their diet could significantly reduce the backfat thickness and perirenal fat percentage of finishing pigs, as well as the drip loss, water-holding capacity, shear force, and the levels of lactate, glucose-6-phosphate, glucose, ATP, phosphofructokinase, and pyruvate in the longissimus dorsi (LD) muscle. It also elevated the levels of flavor amino acids such as glutamate, serine, and threonine, and enriched the composition of flavor substances, including benzene and octanal, which significantly contributed to the enhancement of pork flavor. Furthermore, it enhanced the expression levels of MyHC I and MyHC IIa. In summary, the appropriate addition of FFAI to the diet could improve the carcass traits, meat quality, and antioxidant capacity of finishing pigs. The optimal level of FFAI supplementation is 0.12%. Full article
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<p>Analysis of flavor substances in LD muscle. (<b>A</b>) Electronic nose PCA diagram; (<b>B</b>) radar diagram of odor response values; (<b>C</b>) Cluster heat map of differential flavor substances in LD muscle. Values are the mean ± SEM (<span class="html-italic">n</span> = 8), Significance was expressed as ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Effects of dietary FFAI supplementation on the antioxidant capacity in serum and LD muscle (<b>A</b>), glycolytic potential (<b>B</b>), and muscle fiber types in LD muscle (<b>C</b>) of finishing pigs. CAT, catalase; T-AOC, total antioxidant capacity; GSH-Px, glutathione peroxidase; SOD, superoxide Dismutase; MDA, malondialdehyde, ATP, Adenosine Triphosphate; PFK, Phosphofructokinase; LDH, Lactate Dehydrogenase. Different groups with different lower case letters above the bar graphs indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05). Values are the mean ± SEM (<span class="html-italic">n</span> = 8).</p>
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<p>Correlation analysis between antioxidant capacity, meat quality, glycolysis, and muscle fiber type in finishing pigs based on the Mantel-test at 45min; the redness values of the LD muscle at 45 min; the redness values of the LD muscle at 24 h; LDH, Lactate Dehydrogenase; PFK, Phosphofructokinase. Interaction of antioxidant capacity with meat quality, glycolysis, and muscle fiber type in finishing pigs. Significance was expressed as * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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16 pages, 1281 KiB  
Review
Host Innate and Adaptive Immunity Against African Swine Fever Virus Infection
by Tianqi Zhang, Zixun Lu, Jia Liu, Yang Tao, Youhui Si, Jing Ye, Shengbo Cao and Bibo Zhu
Vaccines 2024, 12(11), 1278; https://doi.org/10.3390/vaccines12111278 - 13 Nov 2024
Viewed by 352
Abstract
Africa swine fever virus (ASFV) is the causative agent of African swine fever (ASF), a highly contagious hemorrhagic disease that can result in up to 100% lethality in both wild and domestic swine, regardless of breed or age. The ongoing ASF pandemic poses [...] Read more.
Africa swine fever virus (ASFV) is the causative agent of African swine fever (ASF), a highly contagious hemorrhagic disease that can result in up to 100% lethality in both wild and domestic swine, regardless of breed or age. The ongoing ASF pandemic poses significant threats to the pork industry and food security, with serious implications for the sanitary and socioeconomic system. Due to the limited understanding of ASFV pathogenesis and immune protection mechanisms, there are currently no safe and effective vaccines or specific treatments available, complicating efforts for prevention and control. This review summarizes the current understanding of the intricate interplay between ASFV and the host immune system, encompassing both innate and adaptive immune responses to ASFV infection, as well as insights into ASFV pathogenesis and immunosuppression. We aim to provide comprehensive information to support fundamental research on ASFV, highlighting existing gaps and suggesting future research directions. This work may serve as a theoretical foundation for the rational design of protective vaccines against this devastating viral disease. Full article
(This article belongs to the Special Issue Adaptive and Innate Response to Viral Disease)
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<p>Innate immune responses against ASFV infection. Upon invading its primary target cells, macrophages, ASFV is recognized by both cell surface and intracellular pattern recognition receptors. This recognition triggers a cascade of signals transmitted through various signaling pathways, activating transcription factors such as NF-κB and IRF3, which regulate the production of IFNs and inflammatory cytokines. When IFNs bind to their receptor complex, they induce multiple downstream signaling pathways, leading to a range of biological effects. The classical STAT1/STAT2 signaling complex interacts with ISRE elements in gene promoters, resulting in the induction of numerous interferon-stimulated genes (ISGs). Additionally, STAT1 homodimers can signal inflammatory responses. The NLRP3 signaling pathway is also activated, leading to pyroptosis in infected cells. During the transcription stage of ASFV infection, ASFV proteins (highlighted in red) inhibit the production of both IFNs and inflammatory cytokines. Notably, different ASFV strains exhibit varying effects on the antigen presentation function of macrophages. Created with BioRender.com.</p>
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<p>Adaptive immune responses during ASFV infection. ASFV has a significant impact on the adaptive immune response, including both humoral and cellular immunity, effectively evading immune clearance and inhibiting immune memory. B and T lymphocytes are the primary cells involved in these responses. B cells, particularly plasma cells, produce virus-specific antibodies—both neutralizing and non-neutralizing—that can neutralize antigens and trigger complement-dependent cytotoxicity as well as antibody-dependent cellular cytotoxicity. Among T cells, CD8+ T cells, including cytotoxic T lymphocytes, and CD4+ CD8+ double-positive (DP) T cells, play crucial roles. CD4+ T cells function as helper T cells, assisting in antigen presentation by antigen-presenting cells. However, ASFV infection leads to a reduction in the overall population of B cells and a decrease in CD4+ and CD8+ T cells. The proportions of DP T cells and CD4+ and CD8+ T cells can vary depending on the virulence of the ASFV strain, with the role of DP T cells during ASFV infection remaining poorly understood. Notably, pigs have a high frequency of circulating γδ T cells, which have been shown to present ASFV antigens to specific T cells in ASFV-immune pigs. However, the functional properties of these γδ T cells warrant further investigation.Created with BioRender.com.</p>
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18 pages, 462 KiB  
Article
Effect of Dietary Xylanase Inclusion on Growth Performance, Nutrient Digestibility, and Digesta Viscosity of Weaned Pigs Fed Wheat–Soybean Meal-Based Diets
by Gabriela M. Galli, Alejandra Forero Salamanca, Keith Haydon, Crystal L. Levesque and Jorge Y. Perez-Palencia
Animals 2024, 14(22), 3255; https://doi.org/10.3390/ani14223255 - 13 Nov 2024
Viewed by 265
Abstract
(1) Background: This study aimed to evaluate the effects of dietary xylanase addition on growth performance, nutrient digestibility, volatile fatty acids, and digesta viscosity at different digestive sites in weaned pigs fed wheat–soybean meal-based diets with reduced metabolizable energy. (2) Methods: A total [...] Read more.
(1) Background: This study aimed to evaluate the effects of dietary xylanase addition on growth performance, nutrient digestibility, volatile fatty acids, and digesta viscosity at different digestive sites in weaned pigs fed wheat–soybean meal-based diets with reduced metabolizable energy. (2) Methods: A total of 312 weaned pigs (5.1 ± 0.9 kg, 20 ± 2 days of age) were assigned to one of six dietary treatments. The experimental diets were formulated in a three-phase nursery feeding program: phase 1 (d0–d7), phase 2 (d8–d21), and phase 3 (d22–d42). The experimental diets consisted of a wheat–soybean meal-based diet formulated to meet pig requirements (positive control, PC); the PC diet with a reduction of 100 kcal of metabolizable energy (ME) (negative control, NC); and the NC diet with either 900, 1800, 3600, or 7200 units of xylanase. Feed disappearance and body weight were measured at d7, 14, 21, and 42 in the nursery phase. The pen fecal score was assessed daily from d0 to d14 and three times a week from d15 to d28. On d21–d24 of the experiment (12 pigs per day), one pig per pen was selected for sample collection: ileal, cecal, and mid-colon digesta for viscosity and ileal digesta, feces for nutrient digestibility, and feces and cecal digesta for the measurement of volatile fatty acid. (3) Results: The addition of xylanase to the NC diets did not improve pig growth performance (body weight, feed conversion ratio, and average daily gain; p > 0.10) during the entire nursery phase. In Week 2 and Week 3, pigs fed xylanase had a lower (χ2 < 0.05) incidence of fecal scores 3 and 4 (diarrhea) than the PC and NC diets. In addition, the apparent total tract digestibility of neutral detergent fiber and acid detergent fiber increased linearly (p < 0.1) in response to xylanase addition. Xylanase addition (900 to 7200 U) decreased digesta viscosity in the colon compared to the PC and NC diets. Furthermore, xylanase addition resulted in a lower (p < 0.05) concentration of acetic, propionic, butyric, valeric, and total volatile fatty acid in cecal samples compared to PC. The addition of xylanase resulted in greater acetic and valeric acid concentrations in cecal samples compared to the NC group (p < 0.10). (4) Conclusions: Xylanase addition can improve nutrient digestibility, particularly at the total tract level, and reduce viscosity in the hindgut, which could be related to decreasing the occurrence of looseness. However, its impact on growth performance was minimal in wheat–soybean meal-based diets with a reduction of 100 kcal of ME. Full article
(This article belongs to the Special Issue Feed Ingredients and Additives for Swine and Poultry)
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<p>Effects of dietary xylanase addition on fecal scores of weaned pigs fed wheat–soybean meal-based diets <sup>1</sup>. <sup>1</sup> Positive control (PC): a wheat/soybean meal-based diet formulated to meet pig requirements (NRC, 2012) except for energy (−50 kcal ME from the requirement to avoid over formulation due to ingredient variation). Negative control (NC): PC diet with reduction of 100 kcal of ME. Xylanase was included at 30, 60, 120, and 240 g/ton in the negative control diets to create 4 additional dietary treatments consisting of xylanase addition at 900, 1800, 3600, and 7200 U/kg feed, respectively (CJ Bio America INC, 2001 Butterfield Road, Suite 720, Downers Grove, IL 60515, USA).</p>
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14 pages, 2457 KiB  
Article
Micro-CT Analysis of Pore Structure in Upland Red Soil Under Different Long-Term Fertilization Regimes
by Huan Fang, Na Zhang, Zhenghong Yu, Dongchu Li, Xinhua Peng and Hu Zhou
Agronomy 2024, 14(11), 2668; https://doi.org/10.3390/agronomy14112668 - 13 Nov 2024
Viewed by 210
Abstract
This study hypothesized that long-term fertilization alters the pore structure and aggregate stability in upland red soil. A long-term fertilization experiment in Qiyang, Hunan, was conducted with three treatments: no fertilization (CK), nitrogen–phosphorus–potassium fertilization (NPK), and nitrogen–phosphorus–potassium combined with pig manure (NPKOM). X-ray [...] Read more.
This study hypothesized that long-term fertilization alters the pore structure and aggregate stability in upland red soil. A long-term fertilization experiment in Qiyang, Hunan, was conducted with three treatments: no fertilization (CK), nitrogen–phosphorus–potassium fertilization (NPK), and nitrogen–phosphorus–potassium combined with pig manure (NPKOM). X-ray computed tomography (CT) scanning technology was used to assess three-dimensional pore structures at both the soil column (50 mm diameter and 50 mm height) and aggregate scales (diameter 3–5 mm), alongside the evaluation of the soil’s physical and chemical properties. Results showed that the soil organic carbon content (SOC) increased by 44.8% in NPK and 112.5% in NPKOM compared to CK. NPKOM improved the aggregate stability by 51.6%, whereas NPK had no significant effect. At the soil column scale, NPK increased the total porosity by 13.7% but reduced larger pores (>0.06 mm), whereas NPKOM decreased the total porosity by 7.8% and increased larger pores. At the aggregate scale, NPKOM increased the porosity for pores >0.098 mm by 7.6 times compared to CK and 9.5 times compared to NPK. In conclusion, long-term NPKOM significantly enhances the SOC and aggregate stability and promotes larger pore formation, unlike NPK, which mainly increases SOC but does not improve the soil structure. Full article
(This article belongs to the Section Soil and Plant Nutrition)
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<p>The location of the experimental site.</p>
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<p>The 2D and 3D characteristics of pores of columns from upland red soils under different fertilizations (27 mm × 27 mm × 27 mm).</p>
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<p>Pore size distribution (<b>a</b>), cumulative porosity (<b>b</b>), Euler number (<b>c</b>) and surface density (<b>d</b>) of upland red soil columns under different fertilizations.</p>
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<p>The 2D characteristics of pores of soil aggregates from upland red soils under different fertilizations.</p>
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<p>Pore size distribution (<b>a</b>), cumulative porosity (<b>b</b>), Euler number (<b>c</b>) and surface density (<b>d</b>) of upland red soil aggregates under different fertilizations.</p>
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<p>Pearson’s correlation coefficients between soil characteristics and soil pore parameters. EN is Euler number and SD is surface density.</p>
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<p>Schematic representation of the arrangement of aggregates and pore distribution within aggregates under different fertilization treatments. (<b>a</b>): the arrangement of aggregates under CK and NPK treatments; (<b>b</b>): the arrangement of aggregates under NPKOM treatments; (<b>c</b>): pore distribution within aggregates under CK and NPK treatments; (<b>d</b>): pore distribution within aggregates under NPKOM treatments.</p>
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31 pages, 63775 KiB  
Article
Microscopic Analysis and Evaluation of Thermal Elevation and Wear of Drills for Implant Site Preparation: An In Vitro Study
by Lucia Memè, Fabrizio Bambini, Tommaso Pizzolante, Martina Principi, Francesco Sampalmieri and Stefano Mummolo
Materials 2024, 17(22), 5524; https://doi.org/10.3390/ma17225524 - 12 Nov 2024
Viewed by 325
Abstract
Drilling for implant site preparation generates heat, which can cause bone necrosis if temperatures exceed 47 °C for over a minute. Factors influencing heat include drill size, speed, pressure, irrigation, and tool wear. Frequent drill replacement is essential, as wear from repeated use [...] Read more.
Drilling for implant site preparation generates heat, which can cause bone necrosis if temperatures exceed 47 °C for over a minute. Factors influencing heat include drill size, speed, pressure, irrigation, and tool wear. Frequent drill replacement is essential, as wear from repeated use and sterilization affects performance. This study compared three pilot drills with similar designs from different manufacturers, testing each on pig ribs for 15 perforations after 15 sterilization cycles. Researchers measured temperature increase, drilling time, and surface wear. Results showed that drill no. 1 generated more heat than drills no. 2 and no. 3, though none reached critical temperatures. Drill no. 2 took the longest to reach the desired depth and displayed the most deformation. Findings highlight the importance of adhering to the recommended operational limits, suggesting that drills should be replaced after 15 cycles to ensure efficacy and patient safety. Full article
(This article belongs to the Special Issue Advanced Materials for Oral Applications)
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<p>Drill no. 1 is an autoclavable stainless steel drill with a diameter of 2.2 mm. It operates at a maximum speed of 800 rpm with irrigation.</p>
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<p>Drill no. 2 is a cylindrical cutter with a diameter of 2 mm, made of carbide-coated stainless steel. Recommended cutting speed: 900–1100 rpm.</p>
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<p>Drill no. 3 is constructed from stainless surgical steel and has a diameter of 2 mm.</p>
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<p>Photos of pilot drill no. 1 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>,<b>c</b>) images of the drill in the middle, (<b>d</b>) image of the drill at the end part.</p>
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<p>Photos of pilot drill no. 1 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>,<b>c</b>) images of the drill in the middle, (<b>d</b>) image of the drill at the end part.</p>
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<p>Photos of pilot drill no. 2 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>,<b>c</b>) images of the drill in the middle, (<b>d</b>) image of the drill at the end part.</p>
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<p>Photos of pilot drill no. 2 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>,<b>c</b>) images of the drill in the middle, (<b>d</b>) image of the drill at the end part.</p>
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<p>Photos of pilot drill no. 3 obtained under the microscope before use. Photos of pilot drill no. 2 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>–<b>d</b>) images of the drill in the middle, (<b>e</b>) image of the drill at the end part.</p>
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<p>Photos of pilot drill no. 3 obtained under the microscope before use. Photos of pilot drill no. 2 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>–<b>d</b>) images of the drill in the middle, (<b>e</b>) image of the drill at the end part.</p>
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<p>Photos of pilot drill no. 3 obtained under the microscope before use. Photos of pilot drill no. 2 obtained under the microscope before use. (<b>a</b>) Image of the drill at tip level, (<b>b</b>–<b>d</b>) images of the drill in the middle, (<b>e</b>) image of the drill at the end part.</p>
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<p>(<b>a</b>–<b>c</b>) Images obtained from the STL file of pilot drill no. 1 pre-use.</p>
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<p>(<b>a</b>–<b>c</b>) Images obtained from the STL file of pilot drill no. 2 pre-use.</p>
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<p>(<b>a</b>–<b>c</b>) Images obtained from the STL file of pilot drill no. 3 pre-use.</p>
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<p>The measurements taken on pilot drills no. 3 (<b>a</b>), no. 2 (<b>b</b>), and no. 1 (<b>c</b>).</p>
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<p>(<b>a</b>–<b>c</b>) Set of the study.</p>
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<p>The initial cutter processed all samples at 1000 rpm, while pilot cutters were tested at 800 rpm for 15 passes with irrigation to a depth of 15 mm.</p>
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<p>A lance cutter drilled 15 holes on each rib, spaced ≥3 mm apart, totaling 45 holes across three ribs. Each sample used a different pilot drill.</p>
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<p>The measurements taken on pilot drills no. 1 (<b>a</b>), no. 2 (<b>b</b>), and no. 3 (<b>c</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 1 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>), and at the end part (<b>d</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 1 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>), and at the end part (<b>d</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 2 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>,<b>d</b>), and at the end part (<b>e</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 2 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>,<b>d</b>), and at the end part (<b>e</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 2 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>,<b>d</b>), and at the end part (<b>e</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 3 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>,<b>d</b>), and at the end part (<b>e</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 3 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>,<b>d</b>), and at the end part (<b>e</b>).</p>
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<p>Photos obtained under the microscope of pilot drill no. 3 (<b>a</b>) after use and sterilization cycles at tip level (<b>b</b>), in the middle (<b>c</b>,<b>d</b>), and at the end part (<b>e</b>).</p>
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<p>(<b>a</b>–<b>c</b>) Images obtained from the STL file of pilot drill no. 1 after use and sterilization cycles.</p>
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<p>(<b>a</b>–<b>c</b>) Images obtained from the STL file of pilot drill no. 2 after use and sterilization cycles.</p>
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<p>(<b>a</b>–<b>c</b>) Images obtained from the STL file of pilot drill no. 3 after use and sterilization cycles.</p>
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<p>Surface comparison reveals a maximum thickness increase of +0.10 mm and a minimum thickness reduction of −0.16 mm, with an average deformation of −0.04 mm across the entire surface analyzed.</p>
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<p>In blue, areas with increased wear are highlighted.</p>
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<p>Surface comparison shows a maximum thickness increase of +0.02 mm, a minimum reduction of −0.15 mm, and an average deformation of −0.05 mm across the analyzed surface.</p>
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<p>In blue, areas with increased wear are highlighted.</p>
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<p>Surface comparison reveals a maximum thickness increase of +0.07 mm, a minimum reduction of −0.15 mm, and an average deformation of −0.03 mm across the analyzed surface.</p>
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<p>In blue, areas with increased wear are highlighted.</p>
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13 pages, 1212 KiB  
Article
Evaluation of the Effects of Electrical and Carbon Dioxide Stunning Methods on Quality Attributes of Pork Meat
by Gábor Jónás, Anna Visy, István Dalmadi, Koppány Majzinger, Nóra Krzyzewsky, Annamária Barkó, József Surányi and László Friedrich
Appl. Sci. 2024, 14(22), 10410; https://doi.org/10.3390/app142210410 - 12 Nov 2024
Viewed by 360
Abstract
In our experiment, the effect of electrical and CO2 stunning on pork meat quality attributes was studied. A total of thirty PIC337 female pigs were allocated to two equal groups which were stunned electrically (50 Hz, 210 V, 2.6 A, 15 s) [...] Read more.
In our experiment, the effect of electrical and CO2 stunning on pork meat quality attributes was studied. A total of thirty PIC337 female pigs were allocated to two equal groups which were stunned electrically (50 Hz, 210 V, 2.6 A, 15 s) or by CO2 (85 V% CO2, 15 V% O2, 90 s) and slaughtered at a commercial slaughterhouse. For 24 h post mortem, the carcass m. gluteus medius (GM) was used to evaluate meat quality attributes such as pH, color, hardness, water holding capacity (WHC), and total pigment content, and meat classification was also carried out. The CO2 stunning resulted in muscle with significantly lower pH (p < 0.01) and poorer water holding capacity (p < 0.05) compared to electrical stunning. The GM of pigs stunned electrically showed significantly increased lightness (L*) and redness (a*) (p < 0.05) compared to CO2-stunned. The time course of development of rigor mortis was similar for both stunning methods. Pale, soft, exudative (PSE) or dark, firm, dry (DFD) meat defects were not observed. Based on total pigment content, stunning methods did not significantly affect the amount of removable blood. Full article
(This article belongs to the Section Food Science and Technology)
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<p>Post mortem pH changes in CO<sub>2</sub>- and electrically stunned pork m. gluteus medius. Data points present the averages and error bars show the standard errors.</p>
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<p>The change in pH as a function of temperature and the spline model predictions for the pH of CO<sub>2</sub>- and electrically stunned pork m. gluteus medius. The solid lines show the ‘ideal’ and ‘cold shortening’ windows.</p>
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<p>The CO<sub>2</sub>- and electrically stunned pork gluteus medius post mortem color changes in lightness (<b>A</b>), redness (<b>B</b>), and yellowness (<b>C</b>). Data points present the averages and error bars show the standard errors.</p>
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<p>The CO<sub>2</sub>- and electrically stunned pork gluteus medius post mortem color changes in lightness (<b>A</b>), redness (<b>B</b>), and yellowness (<b>C</b>). Data points present the averages and error bars show the standard errors.</p>
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<p>Pork m. gluteus medius color difference (ΔE*) between CO<sub>2</sub> and electrical stunning post mortem.</p>
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<p>Changes in pork m. gluteus medius penetration force post mortem in the case of CO<sub>2</sub> and electrical stunning. Data points present the averages and error bars show the standard errors.</p>
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12 pages, 5145 KiB  
Article
Swill and Pig Manure Substrates Differentially Affected Transcriptome and Metabolome of the Black Soldier Fly Larvae
by Bin Zhang, Rencan Yang, Shichun He, Sifan Dai, Qingquan Hu, Xinrong Li, Hongren Su, Jingyi Shi, Zhiyong Zhao and Dongwang Wu
Int. J. Mol. Sci. 2024, 25(22), 12147; https://doi.org/10.3390/ijms252212147 - 12 Nov 2024
Viewed by 271
Abstract
Black soldier fly larvae (Hermetia illucens) (BSFL) are insect larvae with significant ecological and economic value. This study aims to investigate whether swill and manure had any effects on the transcriptome and metabolome of BSFL. Through high-throughput transcriptome sequencing, we found [...] Read more.
Black soldier fly larvae (Hermetia illucens) (BSFL) are insect larvae with significant ecological and economic value. This study aims to investigate whether swill and manure had any effects on the transcriptome and metabolome of BSFL. Through high-throughput transcriptome sequencing, we found that larvae fed with swill exhibited higher levels of gene expression, especially with the upregulation of genes related to energy metabolism, amino acid metabolism, and redox reactions. Metabolomics analysis showed a significant increase in energy metabolism-related metabolites, such as organic acids and amino acids, in the swill-fed larvae. In contrast, gene expression and metabolic characteristics in the pig manure-fed group indicated a higher stress response, with relevant genes and metabolites (such as short-chain fatty acids and antioxidants) showing significant upregulation. GO and KEGG enrichment analyses further supported these results, suggesting that swill promotes larval growth and metabolism, whereas pig manure induces the activation of stress response mechanisms. These findings offer clear molecular and physiological insights into the optimization of substrate selection for enhancing the performance of BSFL in waste management. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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<p>(<b>A</b>) Two-dimensional Principal Component Analysis (PCA) plot. (<b>B</b>) Differential gene clustering heatmap. Note: Each column represents a sample and each row represents a gene. Red indicates relatively high expression of genes, while blue indicates relatively low expression of genes. The dendrogram above represents the clustering of the samples, where the closer two branches are, the more similar the expression patterns of the differential genes in those samples. The dendrogram on the left represents the clustering of the genes, where the closer two gene branches are, the more similar their expression levels. (<b>C</b>) Volcano plot of differentially expressed gene distribution. Note: The horizontal axis represents the fold change in gene expression between different experimental groups or samples, and the vertical axis represents the statistical significance of gene expression changes. Each dot represents a gene, with gray dots indicating non-significantly different genes, red dots indicating significantly upregulated genes, and blue dots indicating significantly downregulated genes.</p>
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<p>(<b>A</b>) Bar chart of GO enrichment for differentially expressed genes. The horizontal axis represents −log10 of the adjusted <span class="html-italic">p</span>-value, with entries arranged in ascending order of <span class="html-italic">p</span>-value. The three main categories of GO (green for biological processes, blue for molecular functions, and red for cellular components) are represented by different colored bars, while the vertical axis describes specific functions. (<b>B</b>) Bubble chart of GO enrichment for differentially expressed genes. The horizontal axis represents −log10 of the adjusted <span class="html-italic">p</span>-value, with entries arranged in ascending order of <span class="html-italic">p</span>-value. The three main categories of GO are shown from top to bottom: biological processes (BPs), molecular functions (MFs), and cellular components (CCs), while the vertical axis provides descriptions of specific functions. (<b>C</b>) Bar chart of KEGG enrichment for differentially expressed genes. The horizontal axis represents −log10 of the adjusted <span class="html-italic">p</span>-value, while the vertical axis displays pathway names. The size of the rich factor is indicated by the color of the bars, and the number of differentially expressed genes within each pathway is shown on the right side of the bars. (<b>D</b>) Bubble chart of KEGG enrichment for differentially expressed genes. The horizontal axis represents −log10 of the adjusted <span class="html-italic">p</span>-value, while the vertical axis shows pathway names. The size of the rich factor is indicated by the color of the bubbles, with larger values indicated by colors closer to red. The size of the bubbles represents the number of differentially expressed genes in each pathway.</p>
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<p>(<b>A</b>) The proportion of identified metabolites in each chemical classification. Note: The different colored blocks in the figure represent various chemical classification categories, and the percentage indicates the proportion of metabolites within that chemical classification relative to the total number of identified metabolites. Metabolites without a defined chemical classification are categorized as undefined. (<b>B</b>) OPLS-DA score plot for positive ion mode. Note: The <span class="html-italic">x</span>-axis [<a href="#B1-ijms-25-12147" class="html-bibr">1</a>] represents principal component 1, while the <span class="html-italic">y</span>-axis [<a href="#B2-ijms-25-12147" class="html-bibr">2</a>] represents principal component 2. Points of the same color represent biological replicates within the same group, and their distribution reflects the differences between and within groups. (<b>C</b>) Analysis of the fold change in expression of significantly different metabolites in positive ion mode. Note: The <span class="html-italic">x</span>-axis indicates the log2 FC values of the differential metabolites, which represent the fold change in these metabolites on a log2 scale. The <span class="html-italic">y</span>-axis represents the significantly different metabolites. Red indicates upregulated differential metabolites, while green indicates downregulated differential metabolites.</p>
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<p>(<b>A</b>) Chord diagram in positive ion mode. Note: The inner circle links represent significant differential metabolites, while the arcs on the outer circle indicate the categories of these metabolites. The colored lines represent correlations within each metabolite category, with lines matching the color of their respective subclasses. Dark gray lines represent correlations between different categories of metabolites. (<b>B</b>) Differential abundance score plot for all differential metabolic pathways. Note: The Y-axis represents the names of the differential pathways, while the X-axis shows the differential abundance score (DA score). The DA score reflects the overall change in all metabolites within a metabolic pathway. A score of 1 indicates an upregulation trend for all identified metabolites in that pathway, while 1 indicates a downregulation trend. The length of the line segment represents the absolute value of the DA score, and the size of the endpoint circles indicates the number of metabolites within that pathway—larger circles signify more metabolites. The color intensity of the lines and circles corresponds to the DA score value, with deeper red indicating a stronger upregulation trend and deeper blue indicating a stronger downregulation trend.</p>
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<p>Swill and pig manure substrates significantly influenced the transcriptome and metabolome of black soldier fly larvae, with the figure showing specific samples before and after treatment at each stage.</p>
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