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Search Results (642)

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15 pages, 788 KiB  
Article
Transcriptomic and Epigenomic Responses to Cortisol-Mediated Stress in Rainbow Trout (Oncorhynchus mykiss) Skeletal Muscle
by Daniela Aravena-Canales, Valentina Valenzuela-Muñoz, Cristian Gallardo-Escarate, Alfredo Molina and Juan Antonio Valdés
Int. J. Mol. Sci. 2024, 25(14), 7586; https://doi.org/10.3390/ijms25147586 (registering DOI) - 10 Jul 2024
Viewed by 163
Abstract
The production and release of cortisol during stress responses are key regulators of growth in teleosts. Understanding the molecular responses to cortisol is crucial for the sustainable farming of rainbow trout (Oncorhynchus mykiss) and other salmonid species. While several studies have [...] Read more.
The production and release of cortisol during stress responses are key regulators of growth in teleosts. Understanding the molecular responses to cortisol is crucial for the sustainable farming of rainbow trout (Oncorhynchus mykiss) and other salmonid species. While several studies have explored the genomic and non-genomic impacts of cortisol on fish growth and skeletal muscle development, the long-term effects driven by epigenetic mechanisms, such as cortisol-induced DNA methylation, remain unexplored. In this study, we analyzed the transcriptome and genome-wide DNA methylation in the skeletal muscle of rainbow trout seven days after cortisol administration. We identified 550 differentially expressed genes (DEGs) by RNA-seq and 9059 differentially methylated genes (DMGs) via whole-genome bisulfite sequencing (WGBS) analysis. KEGG enrichment analysis showed that cortisol modulates the differential expression of genes associated with nucleotide metabolism, ECM-receptor interaction, and the regulation of actin cytoskeleton pathways. Similarly, cortisol induced the differential methylation of genes associated with focal adhesion, adrenergic signaling in cardiomyocytes, and Wnt signaling. Through integrative analyses, we determined that 126 genes showed a negative correlation between up-regulated expression and down-regulated methylation. KEGG enrichment analysis of these genes indicated participation in ECM-receptor interaction, regulation of actin cytoskeleton, and focal adhesion. Using RT-qPCR, we confirmed the differential expression of lamb3, itga6, limk2, itgb4, capn2, and thbs1. This study revealed for the first time the molecular responses of skeletal muscle to cortisol at the transcriptomic and whole-genome DNA methylation levels in rainbow trout. Full article
(This article belongs to the Special Issue Fish Nutrition Program and Epigenetic Regulation)
16 pages, 2337 KiB  
Article
Genetic Characterization of Rhizobium spp. Strains in an Organic Field Pea (Pisum sativum L.) Field in Lithuania
by Justina Kaziūnienė, Francesco Pini, Arman Shamshitov, Kristyna Razbadauskienė, Birutė Frercks, Audrius Gegeckas, Raimonda Mažylytė, Laura Lapinskienė and Skaidrė Supronienė
Plants 2024, 13(14), 1888; https://doi.org/10.3390/plants13141888 - 9 Jul 2024
Viewed by 258
Abstract
Biological nitrogen fixation in legume plants depends on the diversity of rhizobia present in the soil. Rhizobial strains exhibit specificity towards host plants and vary in their capacity to fix nitrogen. The increasing interest in rhizobia diversity has prompted studies of their phylogenetic [...] Read more.
Biological nitrogen fixation in legume plants depends on the diversity of rhizobia present in the soil. Rhizobial strains exhibit specificity towards host plants and vary in their capacity to fix nitrogen. The increasing interest in rhizobia diversity has prompted studies of their phylogenetic relations. Molecular identification of Rhizobium is quite complex, requiring multiple gene markers to be analysed to distinguish strains at the species level or to predict their host plant. In this research, 50 rhizobia isolates were obtained from the root nodules of five different Pisum sativum L. genotypes (“Bagoo”, “Respect”, “Astronaute”, “Lina DS”, and “Egle DS”). All genotypes were growing in the same field, where ecological farming practices were applied, and no commercial rhizobia inoculants were used. The influence of rhizobial isolates on pea root nodulation and dry biomass accumulation was determined. 16S rRNA gene, two housekeeping genes recA and atpD, and symbiotic gene nodC were analysed to characterize rhizobia population. The phylogenetic analysis of 16S rRNA gene sequences showed that 46 isolates were linked to Rhizobium leguminosarum; species complex 1 isolate was identified as Rhizobium nepotum, and the remaining 3 isolates belonged to Rahnella spp., Paenarthrobacter spp., and Peribacillus spp. genera. RecA and atpD gene analysis showed that the 46 isolates identified as R. leguminosarum clustered into three genospecies groups (B), (E) and (K). Isolates that had the highest influence on plant dry biomass accumulation clustered into the (B) group. NodC gene phylogenetic analysis clustered 46 R. leguminosarum isolates into 10 groups, and all isolates were assigned to the R. leguminosarum sv. viciae. Full article
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Figure 1

Figure 1
<p>(<b>A</b>). The average number of nodules on <span class="html-italic">P. sativum</span> roots after inoculation with different rhizobia isolates. Error bars indicate the standard deviations within biological replications at each treatment. Values marked with the same letter are not significantly different at <span class="html-italic">p</span> ≥ 0.05. (<b>B</b>). The averages of the plant dry biomass (columns) and number of nodules on <span class="html-italic">P. sativum</span> roots (ovals) after inoculation with different rhizobia isolate. Error bars indicate the standard deviations within biological replications at each treatment. Values marked with the same letter are not significantly different at <span class="html-italic">p</span> ≥ 0.05.</p>
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<p>Correlation between number of nodules and dry plant biomass, <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>The phylogenetic tree of 16S rRNA gene sequences of the <span class="html-italic">P. sativum</span> root nodule isolates (shown in bold), related strain types of <span class="html-italic">Rhizobium</span> bacteria and reference <span class="html-italic">Bradyrhizobium japonicum</span> LMG 6138; accession numbers indicated in brackets. <span class="html-italic">B. japonicum</span> LMG 6138 was used as an outgroup to root the tree. The evolutionary history was inferred by using the Maximum Likelihood algorithm with the Kimura 2-parameter model. There were a total of 642 positions in the final dataset.</p>
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<p>The phylogenetic tree based on the concatenated alignment of <span class="html-italic">recA</span> and <span class="html-italic">atpD</span> gene sequences of the <span class="html-italic">Rhizobium</span> spp. root nodule isolates (shown in bold) and related strain of <span class="html-italic">Rhizobium</span> bacteria. The evolutionary history was inferred by using the Maximum Likelihood algorithm with the Tamura-Nei model. There were a total of 904 positions in the final dataset. Letters in colored circles indicate the different <span class="html-italic">R. leguminosarum</span> genospecies.</p>
Full article ">Figure 5
<p>The phylogenetic tree of <span class="html-italic">nodC</span> gene sequences of the <span class="html-italic">R. leguminosarum</span> root nodule isolates. The phylogenetic tree is based on the alignment of the <span class="html-italic">nodC</span> gene sequences (from position 337 to 1015 for a total of 679 nucleotides). The tree is inferred using the maximum likelihood algorithm with the Tamura 3 parameter model. Bootstrap values (&gt;50%) are shown next to the branches. Sequences of the rhizobia strains used in this work are highlighted in bold for those strains showing identical <span class="html-italic">nodC</span> sequences; representative sequences for each group have been used.</p>
Full article ">
23 pages, 6045 KiB  
Article
Integrative Taxonomy of Metarhabditis Associated with Parasitic Otitis in Dairy Cattle
by Makoto Enoki Caracciolo, Beatriz Elise de Andrade-Silva, Victor Hugo Borba, Ander Castello-Branco, Hudson Andrade dos Santos, Alena Mayo Iñiguez and Eduardo José Lopes-Torres
Taxonomy 2024, 4(3), 464-486; https://doi.org/10.3390/taxonomy4030023 - 4 Jul 2024
Viewed by 326
Abstract
Parasitic otitis is an inflammatory process that can affect the external to internal cattle ear, causing discomfort in animals, impairing performance, and even leading to animal death. The infection was initially associated with nematodes of the Rhabditis genus in tropical and subtropical regions. [...] Read more.
Parasitic otitis is an inflammatory process that can affect the external to internal cattle ear, causing discomfort in animals, impairing performance, and even leading to animal death. The infection was initially associated with nematodes of the Rhabditis genus in tropical and subtropical regions. Currently, the nematode species described as associated with bovine otitis are Metarhabditis costai, Metarhabditis freitasi, and, more recently, M. blumi. It is worth highlighting that there is still a lack of robust information regarding the morphological details, ultrastructural aspects, and molecular biology data of these species. The Metarhabditis genus is composed of seven species and two more have recently been added. The objective of this study is to update the morphological data using advanced microscopy techniques to emphasize and clarify the main morphological differences between three species of Metarhabditis currently associated with parasitic otitis. Samples of inflammatory exudate were collected from four adult female Gir cattle (Bos taurus indicus) on a farm in Itabira, Minas Gerais state, Brazil. Specimens were analyzed using light microscopy and scanning electron microscopy. Two species, M. costai and M. freitasi, were morphologically identified, consistent with previous reports. Scanning electron microscopy revealed new structural characteristics of the nematode species involved in parasitic otitis compared with M. blumi obtained from the CGC Center. Significant differences were observed in the male posterior region, bursa, and tail. Molecular analysis was conducted to differentiate these three species, and it was observed that the species first associated with otitis formed distinct clusters compared to M. blumi. However, it is important to note that further studies are needed to genetically characterize species of the Metarhabditis genus. Full article
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Figure 1

Figure 1
<p>Male of <span class="html-italic">Metarhabditis costai</span> (<b>A</b>) Entire body, in lateral view; (<b>B</b>) Anterior end, showing the stoma; (<b>C</b>) Posterior end, in ventral view, showing the copulatory bursa.</p>
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<p>Light microscopy of a <span class="html-italic">Metarhabditis costai</span> male. (<b>A</b>) Entire body in ventral view; (<b>B</b>) Anterior end showing the stoma and the anterior segment of the pharynx (<b>C)</b> Posterior end in lateral view; (<b>D</b>) Posterior end in ventral view showing spicules, gubernaculum (*), genital papillae (g), and phasmids (p).</p>
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<p>Scanning electron microscopy of a <span class="html-italic">Metarhabditis costai</span> male. (<b>A</b>) Anterior end showing the oral opening, cephalic papillae (arrows), and amphids (a); (<b>B</b>) Posterior region in lateral view, showing the copulatory bursa, genital papillae (arrows), and spicules; (<b>C</b>) Copulatory bursa, in ventral view, showing the genital papillae distribution (arrows); (<b>D</b>) Detail of the spicule tips, in ventral view.</p>
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<p>Female <span class="html-italic">Metarhabditis freitasi</span>. (<b>A</b>) Entire body in lateral view; (<b>B</b>) Posterior region in lateral view; (<b>C</b>) Egg.</p>
Full article ">Figure 5
<p>Light microscopy of female <span class="html-italic">Metarhabditis freitasi</span>. (<b>A</b>) Entire body in lateral view, showing eggs and larvae; (<b>B</b>) Anterior end, showing the stoma; (<b>C</b>) Detail of the vulva and eggs with embryos at mid-body, in ventral view; (<b>D</b>) Posterior region, lateral view, showing the anus (a), phasmid (p), and with the hair-like tip tail.</p>
Full article ">Figure 6
<p>Scanning electron microscopy of <span class="html-italic">Metarhabditis freitasi</span> female. (<b>A</b>) Anterior end showing the cephalic papillae (arrows) and the amphids (a); (<b>B</b>) Entire body in lateral view; (<b>C</b>) Detail of the vulva in ventral view; (<b>D</b>) Posterior end, showing the anus (arrow) and the tail; (<b>E</b>) Detail of the anus; (<b>F</b>) Detail of the phasmid (p).</p>
Full article ">Figure 7
<p>Male <span class="html-italic">Metarhabditis freitasi</span>. (<b>A</b>) Entire male body, in lateral view; (<b>B</b>) Anterior end, showing the stoma; (<b>C</b>) Posterior end in ventral view.</p>
Full article ">Figure 8
<p>Light microscopy of <span class="html-italic">Metarhabditis freitasi</span> male. (<b>A</b>) Entire body, in ventral view; (<b>B</b>) Anterior end, showing the stoma and the anterior segment of the pharynx; (<b>C</b>) Anterior region showing the bulb and excretory pore opening (e); (<b>D</b>) Posterior region in lateral view; (<b>E</b>) Posterior end in ventral view, showing the copulatory bursa, spicules, and genital papillae; (<b>F</b>) Detailed view of the phasmid (arrow).</p>
Full article ">Figure 9
<p>Scanning electron microscopy of <span class="html-italic">Metarhabditis freitasi</span> male. (<b>A</b>) Anterior end showing the oral opening, cephalic papillae (arrows), and amphids (a); (<b>B</b>) Posterior end, in ventral view, showing the genital papillae (arrows) and spicules of the copulatory bursa; (<b>C</b>) Detailed view of the spicule tips; (<b>D</b>) Detailed view of the phasmid (p) and the tip of the tail in ventral view.</p>
Full article ">Figure 10
<p>Light microscopy of <span class="html-italic">Metarhabditis blumi</span> female. (<b>A</b>) Entire body in ventral view; (<b>B</b>) Anterior end showing the stoma and the anterior end of the pharynx; (<b>C</b>) Vulva (v) and egg (o) in lateral view; (<b>D</b>) Posterior end showing the anus (a); (<b>E</b>) Detail of the tail tip.</p>
Full article ">Figure 11
<p>Scanning electron microscopy of a <span class="html-italic">Metarhabditis blumi</span> female. (<b>A</b>) Anterior end showing the lips and the cephalic papillae (arrows), dorsal (d) and latero-ventral (v); (<b>B</b>) Detail of the vulva at the midbody; (<b>C</b>) Posterior end in ventral view, showing the anus and the phasmids (p); (<b>D</b>) Detail of the anus showing the cuticle elevation.</p>
Full article ">Figure 12
<p><span class="html-italic">Metarhabditis blumi</span>. (<b>A</b>) Anterior region of male; (<b>B</b>) Posterior end of female; (<b>C</b>) Posterior end of male.</p>
Full article ">Figure 13
<p>Light microscopy of a <span class="html-italic">Metarhabditis blumi</span> male. (<b>A</b>) Entire body in ventral view; (<b>B</b>) Anterior end showing the stoma; (<b>C</b>) Posterior end in latero-ventral view, showing the copulatory bursa, the genital papillae distribution (arrows), and the cuticular indentation (r); (<b>D</b>) Detail of the spicule and gubernaculum (arrow); (<b>E</b>) Detail of the cuticular indentation at the posterior end of the copulatory bursa.</p>
Full article ">Figure 14
<p>Scanning electron microscopy of a <span class="html-italic">Metarhabditis blumi</span> male. (<b>A</b>) Anterior end showing the dorsal (d) and latero-ventral (v) lips, cephalic papillae (arrows), and amphids (a); (<b>B</b>) Entire body in lateral view; (<b>C</b>) Posterior end showing the copulatory bursa and genital papillae (arrows). (<b>D</b>) Detail of the spicule; (<b>E</b>) Detail of the phasmids (p) in ventral view and the tip tail.</p>
Full article ">Figure 15
<p>Light microscopy of a <span class="html-italic">Metarhabditis</span> sp. male. (<b>A</b>) Entire body, in lateral view; (<b>B</b>) Posterior end, showing the stoma, (<b>C</b>) Ventral view of the posterior region; (<b>D</b>) Posterior end, in lateral view; (<b>E</b>) Detail of the genital papillae in lateral view; (<b>F</b>) Detail of the spicules and gubernaculum (*); (<b>G</b>) Distribution pattern of the genital papillae (arrows) in ventral view.</p>
Full article ">Figure 16
<p>Scanning electron microscopy of a <span class="html-italic">Metarhabditis</span> sp. male. (<b>A</b>) Anterior end in lateral view showing the dorsal (d) and latero-ventral (v) lips, amphid (a), and cephalic papillae (arrows); (<b>B</b>) Lateral line at the anterior region; (<b>C</b>–<b>E</b>) Sequence of images of the anterior, middle, and posterior regions of the lateral line; (<b>F</b>) Posterior region in laterodorsal view, showing the genital papillae (arrows), the tail, and the projected spicules; (<b>G</b>) Posterior end in ventral view, arrows highlighting the post-cloacal genital papillae; (<b>H</b>) Posterior end, in latero-ventral view, showing the cloacal opening; (<b>I</b>) Detail of the pair of genital papillae (arrows) at the cloacal opening; (<b>J</b>) Detail of the anterior face of the cloacal opening showing a tip-shaped cut-out (arrow).</p>
Full article ">Figure 17
<p>Phylogenetic relationships of <span class="html-italic">Metarhabditis</span> spp. Illustrated by a maximum likelihood phylogram based on the D2/D3 fragment of 28S rDNA. Support values at nodes: aLRT/ML-BP/BPP, respectively.</p>
Full article ">
29 pages, 14554 KiB  
Article
Dietary Chitosan Attenuates High-Fat Diet-Induced Oxidative Stress, Apoptosis, and Inflammation in Nile Tilapia (Oreochromis niloticus) through Regulation of Nrf2/Kaep1 and Bcl-2/Bax Pathways
by Aya G. Rashwan, Doaa H. Assar, Abdallah S. Salah, Xiaolu Liu, Ibrahim I. Al-Hawary, Mohammed H. Abu-Alghayth, Shimaa M. R. Salem, Karim Khalil, Nemany A. N. Hanafy, Alaa Abdelatty, Luyang Sun and Zizy I. Elbialy
Biology 2024, 13(7), 486; https://doi.org/10.3390/biology13070486 - 30 Jun 2024
Viewed by 411
Abstract
Fatty liver injury is a prevalent condition in most farmed fish, yet the molecular mechanisms underpinning this pathology remain largely elusive. A comprehensive feeding trial spanning eight weeks was conducted to discern the potential of dietary chitosan in mitigating the deleterious effects of [...] Read more.
Fatty liver injury is a prevalent condition in most farmed fish, yet the molecular mechanisms underpinning this pathology remain largely elusive. A comprehensive feeding trial spanning eight weeks was conducted to discern the potential of dietary chitosan in mitigating the deleterious effects of a high-fat diet (HFD) while concurrently exploring the underlying mechanism. Growth performance, haemato-biochemical capacity, antioxidant capacity, apoptotic/anti-apoptotic gene expression, inflammatory gene expression, and histopathological changes in the liver, kidney, and intestine were meticulously assessed in Nile tilapia. Six experimental diets were formulated with varying concentrations of chitosan. The first three groups were administered a diet comprising 6% fat with chitosan concentrations of 0%, 5%, and 10% and were designated as F6Ch0, F6Ch5, and F6Ch10, respectively. Conversely, the fourth, fifth, and sixth groups were fed a diet containing 12% fat with chitosan concentrations of 0%, 5%, and 10%, respectively, for 60 days and were termed F12Ch0, F12Ch5, and F12Ch10. The results showed that fish fed an HFD demonstrated enhanced growth rates and a significant accumulation of fat in the perivisceral tissue, accompanied by markedly elevated serum hepatic injury biomarkers and serum lipid levels, along with upregulation of pro-apoptotic and inflammatory markers. In stark contrast, the expression levels of nrf2, sod, gpx, and bcl-2 were notably decreased when compared with the control normal fat group. These observations were accompanied by marked diffuse hepatic steatosis, diffuse tubular damage, and shortened intestinal villi. Intriguingly, chitosan supplementation effectively mitigated the aforementioned findings and alleviated intestinal injury by upregulating the expression of tight junction-related genes. It could be concluded that dietary chitosan alleviates the adverse impacts of an HFD on the liver, kidney, and intestine by modulating the impaired antioxidant defense system, inflammation, and apoptosis through the variation in nrf2 and cox2 signaling pathways. Full article
(This article belongs to the Section Biochemistry and Molecular Biology)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Representative photomicrographs of kidneys from different treatment groups. (<b>A</b>) F6Ch0 shows normal tubular and glomerular structure with moderately vacuolar cytoplasm in addition to adequate interstitial cells. (<b>B</b>) F6Ch5 shows nearly normal histological structure with less vacuolated cytoplasm in addition to slight interstitial cell infiltration (thin arrow). (<b>C</b>) F6Ch10 shows focal tubular degeneration (thin arrow) with interstitial inflammatory aggregations (thick arrow). (<b>D</b>) F12Ch0 shows diffuse tubular damage represented by severe necrotic tubular epithelial cells (thin arrows), intraluminal esinophilic cellular cast (star), and interstitial fibrosis (thick arrow) admixed with lymphocytes and numerous RBCs (arrowhead). (<b>E</b>) F12Ch5 shows mild tubular necrosis (thin arrows). (<b>F</b>) F12Ch10 shows mostly normal tubular endothelial lining with less vacuolation (thin arrows) and little interstitial inflammatory cell infiltration (thick arrow). Scale bar = 50 μm.</p>
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<p>Representative photomicrographs of intestines from different treatment groups. (<b>A</b>) F6Ch0 shows a normal structure of intestinal villi with an intact mucous membrane. (<b>B</b>) F6Ch5 shows normal intestinal mucosa except for a few intestinal vacuoles (thin arrow). (<b>C</b>) F6Ch10 shows mild intestinal vacuolation (thin arrows). (<b>D</b>) F12Ch0 shows shortened intestinal villi with many vacuoles (thin arrow). (<b>E</b>) F123Ch5 shows minimal apical loss (thick arrow) and expanded intestinal mucosa. (<b>F</b>) F12Ch10 shows elongation of the intestinal villi, slight apical mucosal loss (thick arrow), and little intestinal vacuolation (thin arrow). Scale bar = 50 μm.</p>
Full article ">Figure 3
<p>Representative photomicrographs of livers from different treatment groups. (<b>A</b>) F6Ch0 shows normal hepatic architecture with a normal vacuolation appearance. (<b>B</b>) F6Ch5 shows mostly normal hepatocytes and normal lipid vacuoles (thin arrow) with a few slight degenerative changes (thick arrow). (<b>C</b>) F6Ch10 shows hydropic degeneration (thin arrow) with slight blood vessel dilatation. (<b>D</b>) F12Ch0 shows marked hepatocellular necrosis with a loss of most nuclei (thin arrow). (<b>E</b>) F12Ch5 shows relatively vacuolated cytoplasm with intact nuclei. (<b>F</b>) F12Ch10 shows normal hepatocytes with slight degenerative changes in some cells (thick arrow). Scale bar = 50 μm.</p>
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<p>Liver histological morphology in Nile tilapia fed with different experimental diets for 8 weeks. Liver tissue sections were stained with Masson trichrome stain. (<b>A</b>): F6Ch0; (<b>B</b>): F12Ch0 (accumulation of collagen fibers (CFs)); (<b>C</b>): F6Ch5; (<b>D</b>): F6Ch10; (<b>E</b>): F12Ch5 (mild accumulation of collagen fibers (CFs)); (<b>F</b>): F12Ch10. Scale bar = 200 μm.</p>
Full article ">Figure 5
<p>Histological liver sections of Nile tilapia fed with different experimental diets for 8 weeks. Tissue sections were stained with a periodic acid Schiff reagent (PAS) stain. (<b>A</b>): F6Ch0 (accumulated glycogen (AG)); (<b>B</b>): F12Ch0 (slight glycogen (SG)); (<b>C</b>): F6Ch5 (mild glycogen (MG)); (<b>D</b>): F6Ch10 (accumulated glycogen (AG)); (<b>E</b>): F12Ch5 (slight glycogen (SG)); (<b>F</b>): F12Ch10 (mild glycogen (MG)). Scale bar = 200 μm.</p>
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<p>Differential expression of different antioxidant genes in the liver and intestine of Nile tilapia groups fed on normal and high-fat diets with chitosan. (<b>A</b>,<b>B</b>) Nrf2: Nuclear factor erythroid 2-related factor 2, (<b>C</b>,<b>D</b>) Kaep1: Kelch-like ECH-associated protein 1, (<b>E</b>,<b>F</b>) GPx: Glutathione peroxidase, (<b>G</b>,<b>H</b>) SOD: Superoxide dismutase. Columns with different superscript letters in the same figure are significantly different (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Differential expression of apoptosis-related genes in the liver and tight junction-related genes in the intestine of Nile tilapia groups fed on normal and high-fat diets with chitosan. (<b>A</b>) Bax: Bcl-2 associated X-protein, (<b>B</b>) Bcl2: B-cell lymphoma 2, (<b>C</b>) Cyc1: Cytochrome c, (<b>D</b>) Casp3: Cysteine-aspartic acid protease3 in the liver, (<b>E</b>) Casp9: Cysteine-aspartic acid protease9, (<b>F</b>) cldn7: Claudin7, (<b>G</b>) cldn3c: Claudin3c, (<b>H</b>) tjp1: Zonula occludens-1, (<b>I</b>) tjp2: Zonula occludens-2, (<b>J</b>) casp3: Cysteine-aspartic acid protease3 in the intestine. Columns with different superscript letters in the same figure are significantly different (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Differential expression of immune-related genes in the liver and intestine of Nile tilapia groups fed on normal and high-fat diets with chitosan. (<b>A</b>,<b>B</b>) <span class="html-italic">cox2</span>: Cyclooxygenase 2 in the liver and intestine, (<b>C</b>,<b>D</b>) <span class="html-italic">tnf-a</span>: Tumor necrosis factor alpha, (<b>E</b>,<b>F</b>) <span class="html-italic">il-1β</span>: Interleukin-1beta, (<b>G</b>,<b>H</b>) <span class="html-italic">il-10</span>: Interleukin-10. Columns with different superscript letters in the same figure are significantly different (<span class="html-italic">p</span> ≤ 0.05).</p>
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24 pages, 1271 KiB  
Review
Base Characteristics, Preservation Methods, and Assessment of the Genetic Diversity of Autochthonous Breeds of Cattle, Sheep and Pigs in Serbia: A Review
by Radica Djedovic, Dragan Radojkovic, Dragan Stanojevic, Radomir Savic, Natasha Vukasinovic, Mladen Popovac, Vladan Bogdanovic, Cedomir Radovic, Marija Gogic, Nikolija Gligovic, Petar Stojic and Ivan Mitrovic
Animals 2024, 14(13), 1894; https://doi.org/10.3390/ani14131894 - 27 Jun 2024
Viewed by 614
Abstract
Preserving local autochthonous domestic animal populations and the products derived from them is a crucial aspect of managing human utilization of the biosphere. This management approach aims to ensure sustainable benefits for both present and future generations. The diversity of autochthonous domestic animal [...] Read more.
Preserving local autochthonous domestic animal populations and the products derived from them is a crucial aspect of managing human utilization of the biosphere. This management approach aims to ensure sustainable benefits for both present and future generations. The diversity of autochthonous domestic animal populations plays a vital role in the functionality and sustainability of the food production system. It encompasses both productive and non-productive aspects, contributing significantly to the overall health, nutrition, and food security of the landscape by providing a wide range of animal-derived food resources. Based on the data contained in the Draft Program of Rural Development, a significant presence of more than 44 autochthonous and local breeds of domestic animals has been noted in Serbia. In order to enable the sustainable preservation of local domestic animals, the competent Ministry of Agriculture of the Republic of Serbia has, through a number of projects, implemented models for the preservation of local breeds on farms (in situ), as well as provided technical assistance to small farms that keep animal collections. It also helps the local population to procure animals, conducts product quality research, and provides opportunities to integrate conservation programs through tourism. Given that molecular characterization is a key factor for the preservation of autochthonous breeds, in the Republic of Serbia, DNA markers are used for identification and to investigate the belonging to a specific breeds or strain. All the mentioned activities led to an immediate increase in the number of animals, which is especially true for the autochthonous breeds of cattle (Busha), sheep (Sjenicka, Svrljiska, and Vlach-vitohorn) and pigs (Mangalitsa, Moravka, and Resavka) that are discussed in this paper. In addition to the significant measures undertaken to preserve animal genetic resources (AnGR), it is necessary to continue to work primarily on ex situ conservation in order to prevent the loss of their gene pools. However, regardless of the evident effort that has been made to preserve autochthonous genetic resources in Serbia, we believe that there is still a lot of room for further improvement. This primarily refers to advanced technologies that have not been applied so far, mostly related to the identification of genomic regions associated with economic traits, resistance to diseases, and adaptability to emerging climate changes. In this way, the production capacity and functional characteristics of autochthonous species and breeds of domestic animals in Serbia will be improved. Full article
(This article belongs to the Section Animal System and Management)
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<p>Geographical locations of Serbian local breeds of cattle, sheep, and pigs.</p>
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<p>Ways of creating, preserving (in situ and ex situ), and obtaining offspring of high genetic value using different biotechnological methods for improving the traits of interest to breeders.</p>
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13 pages, 2456 KiB  
Article
Identification, Synthesis, and In Vitro Activities of Antimicrobial Peptide from African Catfish against the Extended-Spectrum Beta-Lactamase (ESBL)-Producing Escherichia coli
by Hedmon Okella, Steven Odongo, Didier Vertommen and Emmanuel Okello
Pharmaceutics 2024, 16(7), 850; https://doi.org/10.3390/pharmaceutics16070850 - 22 Jun 2024
Viewed by 914
Abstract
The global surge in multi-drug resistant bacteria, including extended-spectrum β-lactamase (ESBL)-producing Escherichia coli has led to a growing need for new antibacterial compounds. Despite being promising, the potential of fish-derived antimicrobial peptides (AMPs) in combating ESBL-producing E. coli is largely unexplored. In this [...] Read more.
The global surge in multi-drug resistant bacteria, including extended-spectrum β-lactamase (ESBL)-producing Escherichia coli has led to a growing need for new antibacterial compounds. Despite being promising, the potential of fish-derived antimicrobial peptides (AMPs) in combating ESBL-producing E. coli is largely unexplored. In this study, native African catfish antimicrobial peptides (NACAPs) were extracted from the skin mucus of farmed African catfish, Clarias gariepinus, using a combination of 10% acetic acid solvent hydrolysis, 5 kDa ultrafiltration, and C18 hydrophobic interaction chromatography. Peptides were then sequenced using Orbitrap Fusion Lumos Tribrid Mass Spectrometry. The identified peptides were screened for potential antibacterial activity using Random Forest and AdaBoost machine learning algorithms. The most promising peptide was chemically synthesized and evaluated in vitro for safety on rabbit red blood cells and activity against ESBL-producing E. coli (ATCC 35218) utilizing spot-on-lawn and broth dilution methods. Eight peptides ranging from 13 to 22 amino acids with molecular weights between 968.42 and 2434.11 Da were identified. Peptide NACAP-II was non-hemolytic to rabbit erythrocytes (p > 0.05) with a zone of inhibition (ZOI) of 22.7 ± 0.9 mm and a minimum inhibitory concentration (MIC) of 91.3 ± 1.2 μg/mL. The peptide is thus a candidate antibacterial compound with enormous potential applications in the pharmaceutical industry. However, further studies are still required to establish an upscale production strategy and optimize its activity and safety in vivo. Full article
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<p>NACAP-II peptide mass spectrum. The representative MS2 data were sourced out of +4 parent ion. A 433.257 m/z utilized EThcD fragmentation and Orbitrap detection with resolution at 30.000. Peptide, LANVLFRRNATTILQ from protein, W5UKU1 was identified using a series of ions (y-, b-, c-, and z-).</p>
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<p>Predicted peptide 3-D structures of NACAP-II together with respective Ramachandran validation plots. (<b>A</b>) NACAP-II modeled using I-TASSER in stick format. (<b>B</b>) NACAP-II Ramachandran plot modeled using I-TASSER. (<b>C</b>) NACAP-II modeled using PEP-FOLD. (<b>D</b>) NACAP-II Ramachandran plot modeled using PEP-FOLD. Red regions (A, B and L) represent locations of residues in most favored region; Yellow regions (a, b, and l) represent locations of residues in additional allowed regions; Citrine regions (~a, ~b, ~l and ~p) represents locations of residues in generously allowed regions; White regions represents locations of residues in disallowed regions. Most of the amino acid residues in the PEP-FOLD-modeled NACAP-II peptide were in the most favored region (84.6%), with no residue in the disallowed region.</p>
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<p>Chromatogram of the NACAP-II peptide in the 25 min run at 220 nm. Inertsil ODS-SP 4.6 mm × 250 mm reversed-phase high-performance liquid chromatography (RP-HPLC) column with the gradient from solvent A [0.065% Trifluoroacetic acid (TFA) in water] to solvent B (0.05% TFA in Acetonitrile) at a flow rate of 1.0 mL/min.</p>
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<p>Mass spectrum for peptide NACAP-II. The mass spectra for peptide NACAP-II were obtained using a 50% H<sub>2</sub>O/50% MeOH water dissolution method prior to a 0.1 µL injection. Electrospray Ionization (ESI) interface was utilized at +4.5 kV detector. Observed mass was 1730.01, while the theoretical mass was 1730.03.</p>
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<p>Zone of inhibition (ZOI) for NACAP-II peptide on ESBL-producing <span class="html-italic">E. coli</span>. NACAP-II peptide was active with a zone of inhibition of 22.7 ± 0.9 mm. The ESBL-producing <span class="html-italic">E. coli</span> was cultured on Mueller-Hinton Agar (MHA). The experiment was performed in triplicate, and data were expressed as the mean and SEM.</p>
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<p>T-1% Triton X-100. Percentage of hemolysis of rabbit red blood cells via synthesized antimicrobial peptide. The concentration of (1–100) µg/mL did not hemolyze the rabbit blood cells. The experiment was performed in triplicate, and mean values were reported.</p>
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18 pages, 17319 KiB  
Article
Genome-Wide Identification, Expression and Response to Estrogen of Vitellogenin Gene Family in Sichuan Bream (Sinibrama taeniatus)
by Zhe Zhao, Li Peng, Qiang Zhao and Zhijian Wang
Int. J. Mol. Sci. 2024, 25(12), 6739; https://doi.org/10.3390/ijms25126739 - 19 Jun 2024
Viewed by 455
Abstract
To enhance our understanding of teleost reproductive physiology, we identified six Sichuan bream (Sinibrama taeniatus) vitellogenin genes (vtg1-6) and characterized their sequence structures. We categorized them into type Ⅰ (vtg1,4,5 and 6 [...] Read more.
To enhance our understanding of teleost reproductive physiology, we identified six Sichuan bream (Sinibrama taeniatus) vitellogenin genes (vtg1-6) and characterized their sequence structures. We categorized them into type Ⅰ (vtg1,4,5 and 6), type Ⅱ (vtg2) and type Ⅲ (vtg3) based on differences in their subdomain structure. The promoter sequence of vtgs has multiple estrogen response elements, and their abundance appears to correlate with the responsiveness of vtg gene expression to estrogen. Gene expression analyses revealed that the vitellogenesis of Sichuan bream involves both heterosynthesis and autosynthesis pathways, with the dominant pathway originating from the liver. The drug treatment experiments revealed that 17β-estradiol (E2) tightly regulated the level of vtg mRNA in the liver. Feeding fish with a diet containing 100 μg/g E2 for three weeks significantly induced vtg gene expression and ovarian development, leading to an earlier onset of vitellogenesis. Additionally, it was observed that the initiation of vtg transcription required E2 binding to its receptor, a process primarily mediated by estrogen receptor alpha in Sichuan bream. The findings of this study provide novel insights into the molecular information of the vitellogenin gene family in teleosts, thereby contributing to the regulation of gonadal development in farmed fish. Full article
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<p>The gene structure of Sichuan bream vitellogenin. (<b>A</b>) A schematic representation of the genomic structure of <span class="html-italic">vtg1</span>–<span class="html-italic">6</span> (drawn to scale); (<b>B</b>) Promoter sequence analysis diagram. The phylogenetic tree based on <span class="html-italic">vtg</span> promoter sequences is shown on the left, and the corresponding distribution of predicted cis-acting elements is shown on the right.</p>
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<p>Phylogenetic tree based on Vtg polypeptide sequences from Sichuan bream and several other oviparous vertebrates. The right panel displays the YP domains of each Vtg type (not drawn to scale).</p>
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<p>The relative expression of Sichuan bream vitellogenin genes in various tissues.</p>
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<p>The relative expression of Sichuan bream vitellogenin genes at different developmental stages. The relative expression of Sichuan bream vitellogenin genes in the (<b>A</b>) liver and (<b>B</b>) ovary from ovarian development stage Ⅱ to stage Ⅳ. <sup>a, b, c</sup> Mean values with unlike letters were significantly different (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of letrozole (LTZ) and different concentrations of E<sub>2</sub> treatments on the relative expression of Sichuan bream vitellogenin genes in the liver. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; <sup>ns</sup> means values between the comparison groups were no significantly different (<span class="html-italic">p</span> &gt; 0.05).</p>
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<p>Effect of letrozole (LTZ) and different concentrations of E<sub>2</sub> treatments on the ovarian development of Sichuan bream. (<b>A</b>) Gonadosomatic index (GSI) among different groups. (<b>B</b>) Growth parameters among different groups. <sup>a, b, c</sup> Mean values with unlike letters were significantly different (<span class="html-italic">p</span> &lt; 0.05). Ovarian tissue sections corresponding to the (<b>C</b>) control group and under (<b>D</b>) letrozole, (<b>E</b>) E<sub>2</sub> (10 μg/g), (<b>F</b>) E<sub>2</sub> (50 μg/g), (<b>G</b>) E<sub>2</sub> (100 μg/g) and (<b>H</b>) E<sub>2</sub> (200 μg/g) treatments, respectively. Arrows mark cortical alveoli (CA) or yolk granules.</p>
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<p>Effects of SERMs on vitellogenesis of Sichuan bream. (<b>A</b>) The relative expression of the vitellogenin genes in the liver of Sichuan bream after ICI, PPT and DPN treatments; * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01. (<b>B</b>) Gonadosomatic index (GSI) among different groups. (<b>C</b>) Growth parameters among different groups. <sup>a, b</sup> Mean values with unlike letters were significantly different (<span class="html-italic">p</span> &lt; 0.05).</p>
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13 pages, 15265 KiB  
Article
Effects of Soil Microorganisms on Carbon Sequestration under Different Mixed Modification Models in Pinus massoniana L. Plantation
by Meng Chen, Congjun Yuan, Shuang He, Jin Chen, Jun Luo, Fangjun Ding and Guohua Yan
Forests 2024, 15(6), 1053; https://doi.org/10.3390/f15061053 - 18 Jun 2024
Viewed by 459
Abstract
In forests, microbial populations in the soil can directly influence the decomposition of carbon from surface plants, promoting carbon storage and stability. However, in sustainable forest management, it is still unclear how soil microorganisms under different plantation types affect organic carbon sequestration and [...] Read more.
In forests, microbial populations in the soil can directly influence the decomposition of carbon from surface plants, promoting carbon storage and stability. However, in sustainable forest management, it is still unclear how soil microorganisms under different plantation types affect organic carbon sequestration and whether the mechanisms of influence are the same. In this research, we focused on four mixed forests and pure Pinus massoniana-planted forest in the state-owned forest farm of Dushan County. Three replicated plots were set up for each model, and soil samples were collected from different layers (0–20 cm, 20–40 cm, and 40–60 cm), totaling 45 samples. We elucidated the effects of soil microorganisms on carbon sequestration under five mixed modification models of P. massoniana and further explored the mechanisms by which microbial functional communities regulate soil carbon sequestration under different mixed models through molecular sequencing and collinear network analysis. Variance analysis indicated that the soil organic carbon (SOC) of the same soil layer varied significantly, and there were also significant differences in the composition of soil bacterial and fungal microbial communities. Moreover, the bacterial community was more sensitive to changes in the vegetation environment, while the fungal community structure was more resistant to changes in the soil environment. Correlation analysis indicated that the diversity and composition of the bacterial community had more positive effects on soil organic carbon than those of the fungal community. Linear fitting and redundancy analysis (RDA) showed that particulate organic carbon (POC) in soil had the strongest correlation with SOC content. Soil microorganisms affected the storage and stability of soil carbon mainly by regulating the conversion of litter (carbon sources) into POC. The soil environment of different mixed models had different effects on soil carbon accumulation. Both correlation and collinearity network analyses indicated that soil microbial functional groups could enhance carbon storage by regulating readily oxidizable carbon (EOC) and POC content in mixed forest plantations. The results of our study provide a sound basis for replanting a reasonable forest model structure to improve forest carbon storage. Full article
(This article belongs to the Section Forest Soil)
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<p>Location of the test site of the state-owned forest farm in Dushan County.</p>
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<p>Soil carbon component content in different mixing models. A, B, and C indicate significant differences in different mixed planting patterns within the same soil layer, while a indicate significant differences in different soil layers within the same forest model. SOC: soil organic carbon; POC, particulate organic carbon; MBC, microbial biomass C; EOC, readily oxidizable organic carbon; DOC, dissolved organic C; CL, carbon lability.</p>
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<p>Correlation between soil carbon components and SOC in different models.</p>
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<p>Circos plot of soil fungal and bacterial communities in different mixed patterns; (<b>A</b>) relative abundance of phylum levels of fungal communities and (<b>B</b>) that of bacterial communities.</p>
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<p>Box plots of alpha diversity index in soil fungal and bacterial communities. lowercase a, b, c, d indicate significant differences in soil microbial alpha index under different mixed planting modes.</p>
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<p>A 3D plot of soil fungal and bacterial communities based on Bray–Curtis distances.</p>
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<p>Main explanatory modules of fungal and bacterial communities of soil SOC components. (<b>A</b>,<b>B</b>) Collinear network plot of main functional groups of soil microbial community; (<b>C</b>,<b>D</b>) modules of main microbial communities and physicochemical properties of soil and SOC components heatmap. (Note: The X axis is the environmental factor, and the Y axis is the species. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001). Percentage of predominant genera in the top 10 consortia abundance for each module (<b>E</b>). RDA analysis of microbial communities and soil carbon components; TN, total nitrogen; CL, carbon lability.</p>
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14 pages, 8039 KiB  
Article
Variation in the Content and Fluorescence Composition of Dissolved Organic Matter in Chinese Different-Term Rice–Crayfish Integrated Systems
by Ru Liu, Xin Huang, Sujuan Chen, Ying Shi, Lianghu Su, Rongting Ji, Saier Wang, Shentao Zhu, Qifeng Tang and Longjiang Zhang
Sustainability 2024, 16(12), 5139; https://doi.org/10.3390/su16125139 - 17 Jun 2024
Viewed by 345
Abstract
This study examines the fluorescence characteristics of dissolved organic matter (DOM) in soils from different periods of rice–crayfish integrated systems (RCISs) in China. Utilizing three-dimensional excitation–emission matrix (3D-EEM) fluorescence spectroscopy, the study investigated the hydrophobicity, molecular weight distributions, and fluorescence properties of DOM [...] Read more.
This study examines the fluorescence characteristics of dissolved organic matter (DOM) in soils from different periods of rice–crayfish integrated systems (RCISs) in China. Utilizing three-dimensional excitation–emission matrix (3D-EEM) fluorescence spectroscopy, the study investigated the hydrophobicity, molecular weight distributions, and fluorescence properties of DOM in 2-, 5-, and 7-year RCIS operations, with rice monoculture (RM) serving as a control. The findings indicate that in the initial 2 years of an RCIS, factors such as rice straw deposition, root exudates, and crayfish excretions increase dissolved organic carbon (DOC) release and alter DOM composition, increasing the humic acid content in the soil. As the system matures at 5 years, improvements in soil structure and microbial activity lead to the breakdown of high-molecular-weight humic substances and a rise in small-molecular-weight amino acids. By the 7-year mark, as the aquatic ecosystem stabilizes, there is an increase in humic substances and the humification index in the soil DOM. These variations in DOM properties are essential for understanding the effects of integrated farming systems on soil quality and sustainability. Full article
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<p>(<b>a</b>) Locations of the soil sampling sites (Google images of the Earth) in this study. RM: rice monoculture; RCIS-7: 7-year RCIS; RCIS-5: 5-year RCIS; RCIS-2: 2-year RCIS; Schematic of field structure of RCIS: (<b>b</b>) top view, (<b>c</b>) section view.</p>
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<p>The DOC concentrations of the DOM from of the different soil samples.</p>
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<p>The distribution of the hydrophobic/hydrophilic fractions of (<b>a</b>) TOC, (<b>b</b>) polysaccharides, (<b>c</b>) protein; (<b>d</b>) the molecular weight distribution of TOC in soil samples.</p>
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<p>EEM fluorescence spectra of DOM from the soil of RM and different-term RCIS (<b>a</b>) FI, (<b>b</b>) SFI.</p>
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<p>Changes in SUVA among RM and different-term RCIS.</p>
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<p>Changes in Spectral slopes, E2/E3 and SUVA<sub>250–280</sub> among RM and different-term RCIS.</p>
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<p>Four fluorescence DOM compositional indicators—FI<sub>370</sub>, HIX, A:T, β:α—were also calculated as described in the RM and RCIS.</p>
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<p>Stokes shift distributions of the different samples.</p>
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12 pages, 3249 KiB  
Article
The Pathogenic Mechanism of Enterocytozoon hepatopenaei in Litopenaeus vannamei
by Rongrong Ma, Bo Zhu, Jinbo Xiong and Jiong Chen
Microorganisms 2024, 12(6), 1208; https://doi.org/10.3390/microorganisms12061208 - 15 Jun 2024
Viewed by 617
Abstract
Enterocytozoon hepatopenaei (EHP) is a parasite in shrimp farming. EHP mainly parasitizes the hepatopancreas of shrimp, causing slow growth, which severely restricts the economic income of shrimp farmers. To explore the pathogenic mechanism of EHP, the host subcellular construction, molecular biological characteristics, and [...] Read more.
Enterocytozoon hepatopenaei (EHP) is a parasite in shrimp farming. EHP mainly parasitizes the hepatopancreas of shrimp, causing slow growth, which severely restricts the economic income of shrimp farmers. To explore the pathogenic mechanism of EHP, the host subcellular construction, molecular biological characteristics, and mitochondrial condition of Litopenaeus vannamei were identified using transmission electron microscopy (TEM), real-time qPCR, an enzyme assay, and flow cytometry. The results showed that EHP spores, approximately 1 μm in size, were located on the cytoplasm of the hepatopancreas. The number of mitochondria increased significantly, and mitochondria morphology showed a condensed state in the high-concentration EHP-infected shrimp by TEM observation. In addition, there were some changes in mitochondrial potential, but apoptosis was not significantly different in the infected shrimp. The qPCR results showed that the gene expression levels of hexokinase and pyruvate kinase related to energy metabolism were both upregulated in the diseased L. vannamei. Enzymatic activity showed hexokinase and lactate dehydrogenase were significantly increased in the shrimp infected with EHP, indicating EHP infection can increase the glycolysis process and decrease the oxidative phosphorylation process of L. vannamei. Previous transcriptomic data analysis results also support this conclusion. Full article
(This article belongs to the Special Issue Current Insights into Host–Parasite Interactions)
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<p>Shrimp with different concentrations of EHP infection. Note: A is the shrimp sample with a high concentration of EHP infection; B is the shrimp sample with a low concentration of EHP infection.</p>
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<p>Flow chart of glucose metabolism. Note: The text marked in red indicates the enzyme activity detected in this study.</p>
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<p>Histopathological analysis of hepatopancreatic tissues at different concentrations of EHP infection by transmission electron microscopy. Notes: (<b>A</b>,<b>B</b>) represent low-concentration EHP infection groups; (<b>C</b>,<b>D</b>) represent high-concentration EHP infection groups; NC means cell nucleus; red arrows represent EHP; white arrows represent mitochondria.</p>
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<p>HK and PK gene expression in the hepatopancreas of <span class="html-italic">L. vannamei</span>. Notes: HK represents the hexokinase gene; PK represents the pyruvate kinase gene; *** means <span class="html-italic">p</span> ≤ 0.001.</p>
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<p>Enzyme activity analysis in the infected shrimp in the first week. Notes: The control group represents the healthy group (orange ) and the experiment group represents the artificial infection group (blue); ** means <span class="html-italic">p</span> ≤ 0.01, *** means <span class="html-italic">p</span> ≤ 0.001; HK is hexokinase, PK is pyruvate kinase, SDH is succinate dehydrogenase, CCO is cytochrome c oxidase, and LDH is lactate dehydrogenase.</p>
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<p>Enzyme activity analysis in the infected shrimp in the second week. Notes: The relevant annotations are consistent with those in <a href="#microorganisms-12-01208-f005" class="html-fig">Figure 5</a>; ** means <span class="html-italic">p</span> ≤ 0.01, *** means <span class="html-italic">p</span> ≤ 0.001.</p>
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<p>DEGs involved in oxidative phosphorylation pathway in group C vs. group A. Notes: Red indicates upregulated DEGs; dark green represents downregulated DEGs.</p>
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<p>The analysis of cell apoptosis using flow cytometry.</p>
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12 pages, 1470 KiB  
Article
First Molecular Characterisation of Porcine Parvovirus 7 (PPV7) in Italy
by Silvia Dei Giudici, Lorena Mura, Piero Bonelli, Luca Ferretti, Salwa Hawko, Giulia Franzoni, Pier Paolo Angioi, Anna Ladu, Graziella Puggioni, Elisabetta Antuofermo, Maria Luisa Sanna, Giovanni Pietro Burrai and Annalisa Oggiano
Viruses 2024, 16(6), 932; https://doi.org/10.3390/v16060932 - 8 Jun 2024
Viewed by 469
Abstract
Porcine parvoviruses (PPVs) are among the most important agents of reproductive failure in swine worldwide. PPVs comprise eight genetically different species ascribed to four genera: Protoparvovirus (PPV1, PPV8), Tetraparvovirus (PPV2-3), Copiparvovirus (PPV4-6), and Chaphamaparvovirus (PPV7). In 2016, PPV7 was firstly detected in the [...] Read more.
Porcine parvoviruses (PPVs) are among the most important agents of reproductive failure in swine worldwide. PPVs comprise eight genetically different species ascribed to four genera: Protoparvovirus (PPV1, PPV8), Tetraparvovirus (PPV2-3), Copiparvovirus (PPV4-6), and Chaphamaparvovirus (PPV7). In 2016, PPV7 was firstly detected in the USA and afterwards in Europe, Asia, and South America. Recently, it was also identified in Italy in pig farms with reproductive failure. This study aimed to evaluate the circulation of PPV7 in domestic and wild pigs in Sardinia, Italy. In addition, its coinfection with Porcine Circovirus 2 (PCV2) and 3 (PCV3) was analysed, and PPV7 Italian strains were molecularly characterised. PPV7 was detected in domestic pigs and, for the first time, wild pigs in Italy. The PPV7 viral genome was detected in 20.59% of domestic and wild pig samples. PPV7 detection was significantly lower in domestic pigs, with higher PCV2/PCV3 co-infection rates observed in PPV7-positive than in PPV7-negative domestic pigs. Molecular characterisation of the NS1 gene showed a very high frequency of recombination that could presumably promote virus spreading. Full article
(This article belongs to the Special Issue State-of-the-Art Porcine Viruses Research in Europe)
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<p>A maximum likelihood phylogenetic tree inferred from datasets 2 (<b>a</b>), 3 (<b>b</b>), and 4 (<b>c</b>) composed of 61 PPV7 strains by the GTR + G + I model of nucleotide substitution. Isolates under study are indicated with different symbols. Pink triangles: wild boar; green circles: domestic pigs; blue diamonds: free-ranging pigs. Bootstrap values &lt; 70 are not shown. The scale bar indicates the number of substitutions per site.</p>
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12 pages, 925 KiB  
Review
Purpureocillium lilacinum as an Agent of Nematode Control and Plant Growth-Promoting Fungi
by Everlon Cid Rigobelo, Daniel Nicodemo, Olubukola Oluranti Babalola and Nicolas Desoignies
Agronomy 2024, 14(6), 1225; https://doi.org/10.3390/agronomy14061225 - 5 Jun 2024
Viewed by 545
Abstract
Plants support numerous microorganisms within their tissues and the rhizosphere, and these microorganisms, known as the microbiota, can influence plant growth and health. Up to 40% of a plant’s photosynthetic metabolism may be invested in the rhizosphere. The microbiota is considered an extra [...] Read more.
Plants support numerous microorganisms within their tissues and the rhizosphere, and these microorganisms, known as the microbiota, can influence plant growth and health. Up to 40% of a plant’s photosynthetic metabolism may be invested in the rhizosphere. The microbiota is considered an extra genome that can be modulated to meet plant needs. Researchers have identified a set of genes from these microorganisms, known as the microbiome, which can be manipulated to enhance plant growth and health, improve nutrient absorption, reduce the need for chemical fertilizers, increase resistance to pathogens and pests, and increase stress tolerance. In particular, fungi exhibit large genetic and metabolic diversity and are often used to promote plant growth. For example, the fungus Purpureocillium lilacinum has been employed primarily as a biocontrol agent to manage nematodes, but some studies have suggested that it may also promote plant growth by increasing the efficiency of the plant in absorbing nutrients from the soil and providing phytohormones to plants. Therefore, the current review aims to summarize the existing literature on the use of this fungus in agriculture as nematodes control, and discuss its potential as a plant growth-promoter. Full article
(This article belongs to the Section Horticultural and Floricultural Crops)
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<p>Isolation, identification, and characterization of the fungus <span class="html-italic">P. lilacinum</span> from the soil.</p>
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<p>Mode of application of <span class="html-italic">P. lilacinum</span> to crop production.</p>
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14 pages, 3805 KiB  
Article
Prevalence and Genetic Characterization of Porcine Respiratory Coronavirus in Korean Pig Farms
by Ju-Han Kim, Jonghyun Park, Dong-Kyu Lee, Won-Il Kim, Young S. Lyoo, Choi-Kyu Park and Hye-Ryung Kim
Animals 2024, 14(11), 1698; https://doi.org/10.3390/ani14111698 - 5 Jun 2024
Viewed by 458
Abstract
Porcine respiratory coronavirus (PRCV) is a member of the species Alphacoronavirus 1 within the genus Alphacoronavirus of the family Coronaviridae. A few studies have been conducted on the prevalence of PRCV since its first identification in 1997, but there have been no [...] Read more.
Porcine respiratory coronavirus (PRCV) is a member of the species Alphacoronavirus 1 within the genus Alphacoronavirus of the family Coronaviridae. A few studies have been conducted on the prevalence of PRCV since its first identification in 1997, but there have been no recent studies on the prevalence and genetic characterization of the virus in Korea. In this study, the seroprevalence of PRCV was determined in Korean pig farms using a commercially available TGEV/PRCV differential enzyme-linked immunosorbent assay kit. The farm-level seroprevalence of PRCV was determined to be 68.6% (48/70), similar to previous reports in Korea, suggesting that PRCV is still circulating in Korean pig herds nationwide. Among the 20 PRCV-seropositive farms tested in this study, PRCV RNAs were detected in 17 oral fluid samples (28.3%) from nine farms (45.0%), while TGEV RNAs were not detected in any sample. To investigate the genetic characteristics of Korean PRCV strains, genetic and phylogenetic analyses were conducted on PRCV spike gene sequences obtained in this study. The three Korean PRCV strains (KPRCV2401, KPRCV2402, and KPRCV2403) shared 98.5–100% homology with each other and 96.2–96.6% and 91.6–94.5% homology with European and American strains, respectively. A 224-amino acid deletion was found in the S gene of both Korean and European PRCVs but not in that of American PRCVs, suggesting a European origin for Korean PRCVs. Phylogenetic analysis showed that Korean PRCVs are more closely related to European PRCVs than American PRCVs but clustered apart from both, suggesting that Korean PRCV has evolved independently since its emergence in Korean PRCVs. The results of this study will help expand knowledge on the epidemiology and molecular biology of PRCV currently circulating in Korea. Full article
(This article belongs to the Special Issue Studies of Swine Coronavirus)
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<p>Regional distribution of seropositive pig farms for porcine respiratory coronavirus (PRCV) and/or transmissible gastroenteritis virus (TGEV) in Korea. Red circles (<span style="color:red">●</span>), blue circles (<span style="color:blue">●</span>), and green circles (<span style="color:#00B050">●</span>) indicate PRCV antibody-positive, TGEV antibody-positive, and both TGEV and PRCV antibody-positive farms, respectively. The abbreviations GW, GG, CB, CN, GB, GN, JB, JN, and JJ on the map correspond to Gangwon-do, Gyeonggi-do, Chungcheongbuk-do, Chungcheongnam-do, Gyeongsangbuk-do, Gyeongsangnam-do, Jeollabuk-do, Jeollanam-do, and Jeju-do, respectively.</p>
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<p>The pairwise identity matrix based on spike genes of 18 porcine respiratory coronavirus (PRCV) strains and a transmissible gastroenteritis virus (TGEV) reference strain. The matrix illustrates the similarity of nucleotide (nt) and amino acid (aa) sequences between strains based on pairwise comparisons with the spike (S) gene sequences of 18 PRCV strains and a TGEV reference strain. The percent similarities of nt and aa sequences between strains are shown in the lower and upper triangles of the matrix, respectively. The pairwise nt and aa identities of the 19 S gene sequences are color-coded based on the color key provided on the right side of the figure. The TGEV Purdue strain was used as an outgroup.</p>
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<p>Sequence alignment of the N-terminal region of spike genes of 18 porcine respiratory coronavirus (PRCV) strains and a transmissible gastroenteritis virus (TGEV) reference strain. The amino acid (aa) sequence of the TGEV Purdue reference strain is shown in the first line, followed by the sequences of 18 PRCV strains. Sequence numbers are based on the position of aa residues in the S protein sequence of the TGEV Purdue strain. Compared to the sequence of the TGEV Purdue strain, the changed aa residues in the PRCV strains are highlighted in gray. The start and end of deletion regions in the sequences of each PRCV strain are indicated with left-pointing and right-pointing arrows, respectively. The length of the aa deletion region for each PRCV strain is indicated in the middle of the corresponding sequence. The strain names of the three Korean PRCVs are written in bold font.</p>
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<p>Phylogenetic tree of global porcine respiratory coronavirus (PRCV) strains based on the complete spike gene sequences. Black circles indicate three Korean PRCV strains sequenced in this study. The PRCV strains were grouped into two clades: American and European clades. Detailed information about each strain is provided on the right side of the tree. The numbers in each branch represent bootstrap values greater than 50% based on 1000 replicates. Scale bars indicate nucleotide substitutions per site. The transmissible gastroenteritis virus Purdue strain was used as an outgroup.</p>
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13 pages, 7308 KiB  
Article
The Phylogenetic Characterization of Balantioides coli Isolated in the Pavlova Culture Medium Supplemented with Coconut Water and Animal Serum
by Camila Souza Carvalho Class, Laís Lisboa Corrêa, Fabiana Batalha Knackfuss, Maria Regina Reis Amendoeira, Francisco Ponce Gordo and Alynne da Silva Barbosa
Pathogens 2024, 13(6), 476; https://doi.org/10.3390/pathogens13060476 - 4 Jun 2024
Viewed by 323
Abstract
Balantioides coli is a ciliated protist that can cause dysentery in humans, pigs and nonhuman primates and may have the potential for zoonotic transmission. Its diagnosis is routinely performed through conventional parasitological techniques, and few studies have used culturing techniques to isolate it, [...] Read more.
Balantioides coli is a ciliated protist that can cause dysentery in humans, pigs and nonhuman primates and may have the potential for zoonotic transmission. Its diagnosis is routinely performed through conventional parasitological techniques, and few studies have used culturing techniques to isolate it, applying molecular tools for the characterization of this protozoan. Thus, the objective of this study was to confirm B. coli diagnosis using molecular tools and to characterize the genetic variants of this parasite isolated from pigs kept on family farms in Brazil using three different culture media that differed in the serum added. Fecal samples from pigs were inoculated in Pavlova medium plus coconut water (PC), fetal bovine serum (PB) and horse serum (PH). Of the 127 samples positive for forms compatible with the phylum Ciliophora, 31 were selected for isolation. The most successful medium for isolation was PB 19/31 (61.3%), followed by PH 18/31 (58.1%) and PC 11/31 (35.5%). Of the nucleotide sequences generated, 20 were classified as genetic variant type B0, two as A1 and 15 as A0. The results indicated that PC, despite having allowed the isolation of B. coli for a short period, was not an adequate medium for the maintenance of this parasite in vitro, therefore requiring improvement. Full article
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<p>Photos reflecting samples collected after 24 h (1st day), 48 h (2nd day), 72 h (3rd day), 96 h (4th day) and 120 h (5th day) of incubation in an oven for the isolation of ciliated protozoans in modified Pavlova plus coconut water (PC), modified Pavlova plus fetal bovine serum (PB) and modified Pavlova plus horse serum (PH). Photos (<b>A</b>–<b>E</b>) correspond to the isolates in the PC medium; photos (<b>F</b>–<b>J</b>) correspond to the isolates in the PB medium and photos (<b>K</b>–<b>O</b>) correspond to the isolates in the PH medium. Photos (<b>A</b>,<b>F</b>,<b>K</b>) show the 24 h incubation duration; photos (<b>B</b>,<b>G</b>,<b>L</b>) show 48 h; photos (<b>C</b>,<b>H</b>,<b>M</b>) show 72 h; photos (<b>D</b>,<b>I</b>,<b>N</b>) show 96 h and photos (<b>E</b>,<b>J</b>,<b>O</b>) show 120 h. Divisions are highlighted by the tips of the arrows. The photos were taken at 100× magnification; the scale bar represents 150 µm.</p>
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<p>A phylogenetic tree was constructed based on the alignment of 370 base pairs (bp) of the DNA fragment of the rRNA gene and the ITS1-5.8S rRNA-ITS2 fragment from ciliated protozoa using the maximum likelihood method with the TIM2+F+ evolutionary Model G4. The sequences from this study are highlighted in bold and separated by the genetic variant. Sequences of <span class="html-italic">Isotricha prostoma</span> and <span class="html-italic">Troglodytella abrassarti</span> were used as outgroups. The numbers associated with the branches refer to the bootstrap values for 1000 replications.</p>
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17 pages, 4317 KiB  
Article
Transcriptomic Analysis of Sodium-Silicate-Induced Resistance against Rhizoctonia solani AG-3 in Potato
by Yayan Feng, Jianjun Hao, Dongmei Zhang, Hongli Huo, Lele Li, Zhijun Xiu, Chunfang Yang and Xiaoyu Zhang
Agronomy 2024, 14(6), 1207; https://doi.org/10.3390/agronomy14061207 - 3 Jun 2024
Viewed by 214
Abstract
Stem canker and black scurf of potatoes, caused by Rhizoctonia solani, are economically important diseases. Although the field application of sodium silicate has been shown to improve potato’s resistance against R. solani, the underlying mechanism remains unclear. In this study, we [...] Read more.
Stem canker and black scurf of potatoes, caused by Rhizoctonia solani, are economically important diseases. Although the field application of sodium silicate has been shown to improve potato’s resistance against R. solani, the underlying mechanism remains unclear. In this study, we examined this resistance using transcriptomic analysis. Potato stems inoculated with R. solani were treated with sodium silicate, while a control group received no sodium silicate treatment. The plants were grown under natural environmental conditions at the farm of Inner Mongolia Agricultural University. Potato stems were sampled 4, 8, and 12 days after treatment. Total RNA was extracted using the TRIzol reagent and transformed into cDNA. The cDNA was sequenced, the reads were aligned, and the expression levels of genes were quantified and compared between the treated and control groups. A total of 1491 genes were identified as differentially expressed genes (DEGs). Furthermore, these DEGs were found to be involved in hydrolase activity, plant–pathogen interactions, hormone signal transduction, and the phenylpropanoid biosynthesis pathway. To confirm the up- and down-regulation of DEGs, quantitative real-time polymerase chain reaction (qRT-PCR) was performed on randomly selected genes. The results showed that the application of sodium silicate induces a complex defense network in potato plants involving physical barriers, innate immunity, phytohormone signaling, and various phenylpropanoid compounds to combat R. solani infection. This study provides valuable insights into the molecular mechanisms underlying sodium-silicate-induced resistance and its potential for reducing stem canker and black scurf in potato crops. Full article
(This article belongs to the Section Pest and Disease Management)
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<p>Identification of differentially expressed genes (DEGs) between sodium-silicate-treated and non-treated ‘Atlantic’ potato, analyzed using pairwise comparisons of eighteen transcriptomes: (<b>A</b>) The up- and down-regulated DEGs at 4 (SS4-CK4), 8 (SS8-CK8), and 12 (SS12-CK12) days post application (dpa). (<b>B</b>) Venn diagram displaying the distribution of the DEGs at different time points of sodium silicate treatments.</p>
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<p>A heat map of differentially expressed genes (DEGs) between sodium-silicate-treated and non-treated ‘Atlantic’ potato stems at each time point, including 4 (a4 and b4), 8 (a8 and b8), and 12 (a12 and b12) days post application (dpa).</p>
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<p>Gene ontology (GO) term distribution of differential genes for biological process (BP), molecular function (MF), and cellular component (CC) at 4 days post application (dpa) of sodium silicate.</p>
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<p>Gene ontology (GO) term distribution of differential genes for biological process (BP), molecular function (MF), and cellular component (CC) at 8 days post application (dpa) of sodium silicate.</p>
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<p>Gene ontology (GO) term distribution of differential genes for biological process (BP), molecular function (MF), and cellular component (CC) at 12 days post application (dpa) of sodium silicate.</p>
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<p>Differentially expressed genes (DEGs) involved in the plant hormone signal transduction pathway in response to sodium silicate treatment enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis.</p>
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<p>Differentially expressed genes (DEGs) involved in the plant–pathogen interaction pathway in potato with sodium silicate applied based on Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis.</p>
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<p>The relative expression level change of 15 selected genes from DEGs by quantitative real-time PCR. The left vertical coordinate is FPKM (fragments per kilobase of transcript per million mapped reads) of RNA-seq; the right vertical coordinate is the relative expression level of qRT-PCR.</p>
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