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14 pages, 5097 KiB  
Article
Pig and Cow Blood During Cold Storage in CPDA-1 Solution: Hematology and Fluid Behavior
by Ursula Windberger and Andreas Sparer
Biophysica 2025, 5(1), 3; https://doi.org/10.3390/biophysica5010003 - 21 Jan 2025
Viewed by 578
Abstract
Nature equipped red blood cells (RBCs) with diverse mechanical properties, which makes it possible to examine blood with different RBC properties (size, shape, aggregability, deformability). We investigated whether the shelf life of cow blood (stiff RBCs, low aggregability) is longer compared with pig [...] Read more.
Nature equipped red blood cells (RBCs) with diverse mechanical properties, which makes it possible to examine blood with different RBC properties (size, shape, aggregability, deformability). We investigated whether the shelf life of cow blood (stiff RBCs, low aggregability) is longer compared with pig blood (deformability/aggregability comparable to human) due to a delay in RBC clustering and decomposition. Blood was drawn from conscious pigs and cows in their familiar environment to reduce stress and stored 30 days at +7 °C. RBCs remained intact in cow samples whereas pig samples became hemolytic after day 20. White blood cells and platelets decreased with similar percentages in both species. Hematocrit (HCT) decreased due to RBC shrinking in bovine samples and due to RBC decay in porcine samples. Blood viscosity increased in both species although HCT decreased. In porcine samples, shear thinning decreased progressively, indicating a gradual loss of sample cohesion with storage. Yield stress and storage modulus decreased with hemolysis. In HCT-native cow samples, shear thinning, yield stress, and storage modulus showed high intraindividual variability, but the mean values did not change over the time course. In HCT-adjusted (38%) cow samples, solidification occurred after day 7, followed by a reduction in cohesion and shear thinning until the end of storage. Full article
(This article belongs to the Collection Feature Papers in Biophysics)
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Figure 1
<p>Change in the hemograms of porcine (n = 11) and bovine (n = 6) whole blood samples with storage time. (<b>a</b>,<b>b</b>): RBC count and Hb; (<b>c</b>,<b>d</b>): MCV and MCHC; (<b>e</b>,<b>f</b>) WBC and PLT count. Data present mean ± standard deviation of changes relative to baseline.</p>
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<p>Change in the rheological behavior and HCT of porcine (n = 11) and bovine (n = 6) whole blood samples with storage time. Rheological data present mean ± standard deviation of changes relative to baseline. (<b>a</b>,<b>b</b>): Shear viscosity at low and high shear rates. High shear rate viscosity increased continuously in all samples, but the low shear rate viscosity value peaked at day 14 in HCT-adjusted bovine samples, representing blood thickening; (<b>c</b>): shear thinning (η<sub>10</sub>/η<sub>1000</sub>) decreased in porcine and HCT-adjusted bovine samples towards the end of the observation period as an indicator of a deteriorating suspension but not in HCT-native bovine samples. The significant thickening of HCT-adjusted bovine samples is also reflected in the rise of shear thinning on day 14. Due to the species-specific difference, the HCT is displayed as absolute values. Technical problems prevented rheometry of HCT-native bovine samples on day 0. (<b>d</b>): Change in HCT in the form of absolute values.</p>
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<p>Blood smears of porcine and three selected bovine blood samples at the beginning and the end of storage. After 30 days of storage: pig 1: ghosts (red arrows) and cell debris; pig 2: crenated cells (green arrows); pig 3: ghosts. After 30 days of storage: cow 1 and 3: regular round shapes; cow 2: crenated cells. Scale bar: 20 μm.</p>
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<p>Yield points of bovine (n = 6, blue) and porcine (n = 11, green) whole blood obtained by amplitude sweep tests. (<b>a</b>,<b>b</b>): change in yield point with storage duration; the boxes represent median and interquartile range, asterisks show the mean value. (<b>c</b>,<b>d</b>): intraindividual variability of yield points during the time course; (<b>e</b>,<b>f</b>): yield point of fresh blood and aged blood on the 30th storage day: a quadratic regression curve interpolates the G′-values. The yield stress was obtained from the crossing point of the tangent that was drawn on the inflection point of this regression curve and crossed with a horizontal line through the first G′-values, which was extrapolated to the x-axis (method described in [<a href="#B12-biophysica-05-00003" class="html-bibr">12</a>]).</p>
Full article ">Figure 5
<p>(<b>a</b>,<b>b</b>): Frequency spectrum of G′ and G″ of cow (n = 6, blue) and pig (n = 11, green) whole blood at start and end of storage. Pig blood fluidified, as indicated by the decrease in G′, whereas the shear moduli of HCT-native bovine blood hardly altered. (<b>c</b>,<b>d</b>): The intraindividual variability of loss factor (G″/G′) during the time course. The inset in <a href="#biophysica-05-00003-f002" class="html-fig">Figure 2</a>c shows the HCT-adjusted bovine sample. (<b>e</b>,<b>f</b>): The change in loss factor with storage duration. In porcine samples, loss factor values increased beyond day 22 due to hemolysis. In HCT-native bovine samples, loss factor values showed large errors but did not change with storage time (except one outlier at +24 days, cow E). In HCT-adjusted bovine samples, the loss factor decreased transiently around day 14 and returned afterwards. Boxes represent median and interquartile range; asterisks show the mean value. Values below the torque limit of the rheometer (1 μNm) are deleted from the spectrum in <a href="#biophysica-05-00003-f005" class="html-fig">Figure 5</a>a.</p>
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<p>Blood smear from pig 1 (a different window of this smear is also shown in <a href="#biophysica-05-00003-f003" class="html-fig">Figure 3</a>) showing the clusters of cell debris and free hemoglobin.</p>
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18 pages, 319 KiB  
Article
Impact of Agroindustrial Waste Fermented with Bacteria and Yeasts and Their Effect on Productive, Hematological, and Microbiota Indicators in Guinea Pigs (Cavia porcellus)
by José E. Miranda-Yuquilema, Juan Taboada-Pico, Daniel Luna-Velasco, Mercy Cuenca-Condoy and Wilfrido Briñez
Fermentation 2025, 11(1), 10; https://doi.org/10.3390/fermentation11010010 - 2 Jan 2025
Viewed by 532
Abstract
In the last decade, the production of guinea pig meat in Andean countries has increased due to the growing number of consumers of this meat. Objective: To evaluate the effect of including different doses (0.50, 1.00, and 1.50 mL) of agro-industrial substrates (molasses [...] Read more.
In the last decade, the production of guinea pig meat in Andean countries has increased due to the growing number of consumers of this meat. Objective: To evaluate the effect of including different doses (0.50, 1.00, and 1.50 mL) of agro-industrial substrates (molasses distillery waste) fermented with lactic acid bacteria and yeasts on productive performance, hematological profile, relative weight changes in digestive tract organs, and changes in the intestinal microbiota in guinea pigs (Cavia porcellus). Materials: A total of 300 guinea pigs, Kuri breed, aged 20 days and weighing 330 g, were distributed into 10 groups of 30 animals each. Ctrl, Control. La, substrate fermented with Lactobacillus acidophilus (8.1 × 107 CFU/mL). Kf, substrate fermented with Kluyveromyces fragilis (7.4 × 106 CFU/mL). La + Kf, substrate fermented with bacteria and yeasts; the evaluated doses were 0.50, 1.00, and 1.50 mL/animal. The indicators evaluated in the study included weight gain, health, hematological profile, relative weight of digestive tract organs, and changes in the intestinal microbiota. Results: The parameters evaluated were toxicity, productive parameters, occurrence of diarrhea and mortality, and blood profile. The results showed a significant increase in the weight of the animals consuming probiotics, especially at higher doses. Additionally, an improvement in the intestinal microbiota was observed, with an increase in beneficial bacteria such as Lactobacillus and a decrease in pathogenic bacteria. Probiotics also influenced the hematological parameters and the weight of digestive tract organs, suggesting a positive effect on the overall health of the animals. Conclusions: Supplementation with probiotics proved to be a promising strategy for improving productive performance and intestinal health in guinea pigs. Supplementation with L. acidophilus and K. fragilis significantly enhances guinea pig growth and modulates the intestinal microbiota. The combination of strains and appropriate doses maximizes benefits. These results promise applications in animal production, requiring further studies to confirm their efficacy in other species and developmental stages. Full article
14 pages, 1902 KiB  
Article
Genomic Variants Associated with Haematological Parameters and T Lymphocyte Subpopulations in a Large White and Min Pig Intercross Population
by Naiqi Niu, Runze Zhao, Ming Tian, Wencheng Zong, Xinhua Hou, Xin Liu, Ligang Wang, Lixian Wang and Longchao Zhang
Animals 2024, 14(21), 3140; https://doi.org/10.3390/ani14213140 - 1 Nov 2024
Viewed by 908
Abstract
The breeding of disease-resistant pigs has consistently been a topic of significant interest and concern within the pig farming industry. The study of pig blood indicators has the potential to confer economic benefits upon the pig farming industry, whilst simultaneously providing valuable insights [...] Read more.
The breeding of disease-resistant pigs has consistently been a topic of significant interest and concern within the pig farming industry. The study of pig blood indicators has the potential to confer economic benefits upon the pig farming industry, whilst simultaneously providing valuable insights that can inform the study of human diseases. In this study, an F2 resource population of 489 individuals was generated through the intercrossing of Large White boars and Min pig sows. A total of 17 haematological parameters and T lymphocyte subpopulations were measured, including white blood cell count (WBC), lymphocyte count (LYM), lymphocyte count percentage (LYM%), monocyte count (MID), monocyte count percentage (MID%), neutrophilic granulocyte count (GRN), percentage of neutrophils (GRN%), mean platelet volume (MPV), platelet distribution width (PDW), platelet count (PLT), CD4+/CD8+, CD4+CD8+CD3+, CD4+CD8−CD3+, CD4−CD8+CD3+, CD4−CD8−CD3+, and CD3+. The Illumina PorcineSNP60 Genotyping BeadChip was obtained for all of the F2 animals. Subsequently, a genome-wide association study (GWAS) was conducted using the TASSEL 5.0 software to identify associated variants and candidate genes for the 17 traits. Significant association signals were identified for PCT and PLT on SSC7, with 1 and 11 significant SNP loci, respectively. A single nucleotide polymorphism (SNP) on SSC12 was identified as a significant predictor of the white blood cell (WBC) trait. Significant association signals were detected for the T lymphocyte subpopulations, namely CD4+/CD8+, CD4+CD8+CD3+, CD4+CD8−CD3+, and CD4−CD8+CD3+, with the majority of these signals observed on SSC7. The genes CLIC5, TRIM15, and SLC17A4 were identified as potential candidates for influencing CD4+/CD8+ and CD4−CD8+CD3+. A missense variant, c.2707 G>A, in the SLC17A4 gene has been demonstrated to be significantly associated with the CD4+/CD8+ and CD4-CD8+CD3+ traits. Three missense variants (c.425 A>C, c.500 C>T, and c.733 A>G) have been identified in the TRIM15 gene as being linked to the CD4+/CD8+ trait. Nevertheless, only c.425 A>C has been demonstrated to be significantly associated with CD4-CD8+CD3+. In the CLIC5 gene, one missense variant (c.957 T>C) has been identified as being associated with the CD4+/CD8+ and CD4-CD8+CD3+ traits. Additionally, significant association signals were observed for CD4+CD8+CD3+ and CD4+CD8−CD3+ on SSC2 and 5, respectively. Subsequently, a gene ontology (GO) enrichment analysis was conducted on all genes within the quantitative trait loci (QTL) intervals of platelet count, CD4+/CD8+, and CD4−CD8+CD3+. The MHC class II protein complex binding pathway was identified as the most significant pathway among the three immune traits. These results provide guidance for further research in the field of breeding disease-resistant pigs. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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<p>Identification of GWAS of the haematological parameters. (<b>A</b>) Manhattan plot displaying the GWAS results of plateletocrit (PCT). The blue horizontal line indicated the Bonferroni significance threshold (1.17 × 10<sup>−6</sup>). (<b>B</b>) Manhattan plot displaying the GWAS results of the platelet count (PLT). (<b>C</b>) Manhattan plot displaying the GWAS results of white blood cell count (WBC).</p>
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<p>Comparison of three genotypes of significant SNP for haematological traits. (<b>A</b>) The difference analysis of MARC0014928 for PCT in SSC7. (<b>B</b>) The difference analysis of MARC0014928 for PLT in SSC7. (<b>C</b>) The difference analysis of ASGA0098229 for WBC in SSC12. (* <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, **** <span class="html-italic">p</span> &lt; 0.0001, (<span class="html-italic">p</span> &gt; 0.05, ns)).</p>
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<p>Identification of GWAS of the T lymphocyte subpopulation. (<b>A</b>) Manhattan plot displaying the GWAS results of the CD4+/CD8+. The blue horizontal line indicated the Bonferroni significance threshold (1.17 × 10<sup>−6</sup>). (<b>B</b>) Manhattan plot displaying the GWAS results of the CD4+CD8+CD3+. (<b>C</b>) Manhattan plot displaying the GWAS results of the CD4+CD8−CD3+. (<b>D</b>) Manhattan plot displaying the GWAS results of the CD4−CD8+CD3+.</p>
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<p>Comparison of three genotypes of significant SNP in T lymphocyte subpopulation traits. (<b>A</b>) The difference analysis of ASGA0031860 for CD4+/CD8+ in SSC7. (<b>B</b>) The difference analysis of ALGA0017071 for CD4+CD8+CD3+ in SSC2. (<b>C</b>) The difference analysis of ASGA0032099 for CD4+CD8−CD3+ in SSC7. (<b>D</b>) The difference analysis of ASGA0031860 for CD4−CD8+CD3+ in SSC7. (* <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, **** <span class="html-italic">p</span> &lt; 0.0001, (<span class="html-italic">p</span> &gt; 0.05, ns)).</p>
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<p>Bubble chart of GO function enrichment analysis of genes in SSC7. (<b>A</b>) PLT of GO function enrichment analysis. (<b>B</b>) CD4+/CD8+ of GO function enrichment analysis. (<b>C</b>) CD4−CD8+CD3+ of GO function enrichment analysis.</p>
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16 pages, 3300 KiB  
Article
Experimental Infections of Pigs with Japanese Encephalitis Virus Genotype 4
by Paul M. Hick, Deborah S. Finlaison, Kate Parrish, Xingnian Gu, Philip Hayton, Tiffany O’Connor, Andrew Read, Jing Zhang, Zoe B. Spiers, Pedro Pinczowski, Angel L. Ngo and Peter D. Kirkland
Microorganisms 2024, 12(11), 2163; https://doi.org/10.3390/microorganisms12112163 - 26 Oct 2024
Viewed by 1131
Abstract
The emergence of Japanese encephalitis virus (JEV) in eastern Australia in 2022 caused extensive reproductive disease in pigs and is a threat to public health. Groups of weaned piglets were experimentally infected with the Australian outbreak strain of JEV (genotype 4). All pigs [...] Read more.
The emergence of Japanese encephalitis virus (JEV) in eastern Australia in 2022 caused extensive reproductive disease in pigs and is a threat to public health. Groups of weaned piglets were experimentally infected with the Australian outbreak strain of JEV (genotype 4). All pigs challenged at 5 weeks of age were infected after an intradermal injection of 1 × 105.5 (n = 4) or 1 × 104.5 TCID50/pig (n = 5). Intranasal instillation was less effective at this age, infecting 3/4 pigs with the same higher dose and 1/5 with the lower dose. Intradermal injection using 1 × 105.0 TCID50/pig also infected 9/9 pigs at 11 weeks of age. Infection in all cases was confirmed by qRT-PCR of blood samples, which identified a viremia peak at 3–4 days and detected JEV-specific antibodies as early as 5 days after the challenge. The detection of JEV in oral and nasal swabs and in saliva from chew ropes was less consistent. JEV was detected in the tonsils of 21/22 infected pigs and was isolated from the tonsils of 9/9 pigs sampled 19 days after the challenge at 11 weeks of age. The infected pigs showed no clinical signs other than pyrexia on Days 4–6. Histopathology consistent with JEV infection was evident in the nervous tissues of all but two pigs sampled 28 days after the challenge and was characterized by meningitis, encephalitis and gliosis throughout the brain. Serological studies showed extensive cross-reactivity between JEV and Murray Valley encephalitis virus using blocking ELISAs. However, the determination of limiting-dilution titres allowed for the identification of the infecting virus. This in vivo infection model will be useful in evaluating JEV vaccines and for comparative pathogenesis studies with other JEV genotypes. Full article
(This article belongs to the Special Issue Viral Infection in Animals)
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<p>Rectal temperature of pigs measured daily after JEV challenge. Data are the mean and standard deviation; significant differences between infected and uninfected pigs on each day are indicated (<span class="html-italic">p</span> &lt; 0.01) *. (<b>a</b>) Pigs challenged at 5 weeks of age were classified as infected or not infected based on the composite outcome of all laboratory analyses. There were 13 pigs that were infected by intradermal or intranasal challenge and 14 that were not infected (9 negative controls and 5 challenged by intranasal inoculation that showed no evidence of infection). (<b>b</b>) Pigs challenged at 11 weeks of age by intradermal injection were all infected (<span class="html-italic">n</span> = 9). Whilst there were no significant differences between infected and uninfected groups on any day, 3 of 9 infected pigs on Day 4 had pyrexia (temperature &gt; 40.0 °C).</p>
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<p>Histopathological lesions in the central nervous system of pigs infected with JEV at 5 weeks of age and sampled 10 days after challenge. (<b>a</b>) Brainstem section showing necrotising encephalitis. Focal extensive area of necrosis (malacia) adjacent to a lymphoplasmacytic perivascular cuffing, associated with moderate infiltration of macrophages, lymphocytes, plasma cells and a few neutrophils; HE, 20×. (<b>b</b>) Cerebrum section showing lymphoplasmacytic meningitis. Meninges are expanded due to moderate infiltration of macrophages, lymphocytes and plasma cells; HE, 20×. (<b>c</b>) Cervical spinal cord section showing lymphoplasmacytic myelitis. Mild to moderate focal infiltrate of lymphocytes, macrophages and plasma cells; HE, 40×. (<b>d</b>) Cerebrum section showing moderate lymphoplasmacytic encephalitis. Three vessels are cuffed by up to 3 layers of lymphocytes and plasma cells; HE, 10×.</p>
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<p>Histopathological lesions in tissues of pigs infected with JEV at 11 weeks of age and sampled 19 days after challenge. (<b>a</b>) Prostate section showing lymphoplasmacytic and neutrophilic prostatitis. Mild to moderate infiltrates of lymphocytes, plasma cells, macrophages and a few neutrophils surrounding prostatic glands. Glands are occasionally dilated and filled with eosinophilic fluid and a few necrotic cells. HE, 10×. (<b>b</b>) Kidney section showing lymphoplasmacytic interstitial nephritis. Mild to moderate infiltrates of lymphocytes, plasma cells and macrophages associated with mild fibrosis and surrounding tubules; HE, 10×.</p>
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19 pages, 3461 KiB  
Article
Monoclonal Antibody against Porcine LAG3 Inhibits Porcine Reproductive and Respiratory Syndrome Virus Infection
by Hui Wang, Xu Zheng, Danyang Zheng, Xiaoqian Wang, Zhiqian Zhao, Mi Zhao, Qiang Guo and Yang Mu
Vet. Sci. 2024, 11(10), 483; https://doi.org/10.3390/vetsci11100483 - 7 Oct 2024
Viewed by 1659
Abstract
Lymphocyte activation gene 3 (LAG3) is an inhibitory receptor and the interaction between fibrinogen-like protein 1 and LAG3 can inhibit the anti-tumor effect of T cells both in vivo and in vitro, which was regarded as a new immune evasion mechanism. Porcine reproductive [...] Read more.
Lymphocyte activation gene 3 (LAG3) is an inhibitory receptor and the interaction between fibrinogen-like protein 1 and LAG3 can inhibit the anti-tumor effect of T cells both in vivo and in vitro, which was regarded as a new immune evasion mechanism. Porcine reproductive and respiratory syndrome (PRRS), caused by PRRSV, is an infectious disease characterized by reproductive disorders in pregnant sows and gilts and respiratory problems in pigs of all ages, seriously impacting the pig industry worldwide. In this study, monoclonal antibodies (mAbs) against porcine LAG3 (pLAG3) were developed, and one mAb (1C2) showed good reactivity with pLAG3 on PHA-activated porcine peripheral blood lymphocytes. Epitope mapping showed the epitope recognized by mAb 1C2 was located at amino acid residues 214–435 of pLAG3. LAG3 expression in the tissues of PRRSV-infected pigs was detected, using mAb 1C2 as the primary antibody, and the results revealed that PRRSV infection caused a marked increase in LAG3 expression compared to the control group. Interference of LAG3 expression on PHA-activated lymphocytes promoted PRRSV replication in the co-culture system of monocyte-derived dendritic cells and lymphocytes, whereas overexpression of LAG3 or blocking of the LAG3 signal with mAb 1C2 inhibited PRRSV replication, indicating that PRRSV infection activates the LAG3-signaling pathway, suggesting that this pathway plays an important role in PRRSV pathogenesis. The results obtained lay the foundation for subsequent research on the role of LAG3 in PRRS and other diseases with persistent infection characteristics. Full article
(This article belongs to the Special Issue Emerging and Re-emerging Swine Viruses)
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<p>Amplification, expression, identification, and purification of recombinant <span class="html-italic">pLAG3</span>. (<b>A</b>) Amplified <span class="html-italic">pLAG3</span> (109–1305 nt) gene. M, DL8000 DNA marker; 1–3, <span class="html-italic">pLAG3</span> gene. (<b>B</b>) Identify <span class="html-italic">pLAG3</span> expression by SDS-PAGE and (<b>C</b>) Western blotting analysis. M, PageRuler Prestained Protein Ladder; 1, no-induced BL21-pET28a-pLAG3; 2, induced BL21-pET28a-pLAG3; 3, supernatant after lysis; 4, precipitate after lysis. (<b>D</b>) Purification of <span class="html-italic">pLAG3</span>. M, PageRuler Prestained Protein Ladder; 1, sample loaded; 2, effluent; 3, purified <span class="html-italic">pLAG3</span>.</p>
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<p>Characteristics of <span class="html-italic">pLAG3</span> mAbs. (<b>A</b>) Titers of mouse sera antibody against <span class="html-italic">pLAG3</span> detected by iELISA. Using anti-His mAb as a positive control, naive mouse serum as a negative control, mAb 1C2 or 3E11 as primary antibody, the reaction of mAbs 1C2 and 3E11 with <span class="html-italic">pLAG3</span> expressed by HEK-293T cells was determined by Western blotting (<b>B</b>) and indirect immunofluorescent assay (IFA) (<b>C</b>). (<b>D</b>) Titers of mAbs 1C2 and 3E11 in ascites detected by iELISA. (<b>E</b>) Isotype of mAbs 1C2 and 3E11 determined with an IsoStrip<sup>TM</sup> Mouse Monoclonal Antibody Isotyping Kit (Cat# 11493027001, Roche). (<b>F</b>) Saturation curves of mAbs 1C2 and 3E11 with <span class="html-italic">pLAG3</span>. Per-well plates were coated with 200 ng purified recombinant <span class="html-italic">pLAG3</span>; mAbs 1C2 and 3E11 diluted differently were used to perform iELISA, and mAb2-5G2 was used as a negative control. (<b>G</b>) Purified mAb 1C2 identified by SDS-PAGE.</p>
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<p>Identification of the antigen region recognized by mAb 1C2. (<b>A</b>) Schematic diagram of <span class="html-italic">pLAG3</span> fragments. (<b>B</b>) Procaryotic expression of truncated <span class="html-italic">pLAG3</span> and the reaction of mAb 1C2 with the truncated <span class="html-italic">pLAG3</span> identified by Western blotting. Porcine HEV 239 protein is a control protein expressed with the same vector and <span class="html-italic">E. coli</span> cells. (<b>C</b>) pLAG3-3 expressed by HEK-293T cells and the reaction of mAb 1C2 with pLAG3-3 identified by Western blotting.</p>
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<p>PRRSV infection promotes LAG3, SLA-DRα, and FGL1 expression in vivo. (<b>A</b>) PRRSV load, (<b>B</b>) <span class="html-italic">pLAG3</span>, (<b>D</b>) SLA-DRα, and (<b>F</b>) FGL1 gene expression were detected by RT-qPCR. The total RNA of lung, liver, spleen, and MLN samples from mock and PRRSV-infected pigs were extracted. Serial 10-fold dilutions of a recombinant plasmid containing 144 bp PRRSV ORF7 gene were introduced in each plate to detect inter-run variations, and the PRRSV load was shown as quantification cycle (Cq) value. Gene expressions were calculated with the 2<sup>−ΔΔCt</sup> method, and GAPDH was used as an internal control. The results are shown as mean ± SD from three independent experiments. NS means no significant difference, * <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, **** <span class="html-italic">p</span> &lt; 0.0001. (<b>C</b>) <span class="html-italic">pLAG3</span>, (<b>E</b>) SLA-DRα, and (<b>G</b>) FGL1 protein expressions in different tissues were detected by Western blotting, using homemade mAbs 1C2, 2E11D9, or 4G7 as the primary antibody.</p>
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<p>Induction of MoDCs in vitro. (<b>A</b>) Changes in cell morphology during MoDCs induction under a light microscope. (<b>B</b>) SLA-DRα expression was detected by Western blotting using homemade mAb 2E11D9. (<b>C</b>) PRRSV viral load was detected by RT-qPCR with 1 MOI PRRSV SD16 infection at 24 h. Serial 10-fold dilutions of a recombinant plasmid containing 144 bp PRRSV ORF7 gene were introduced in each plate to detect inter-run variations, and the PRRSV load was shown as quantification cycle (Cq) value. Gene expressions were calculated with the 2<sup>−ΔΔCt</sup> method, and GAPDH was used as an internal control. The results are shown as mean ± SD from three independent experiments. *** <span class="html-italic">p</span> &lt; 0.001. (<b>D</b>) PRRSV N protein expression was detected by Western blotting, using homemade mAbs 6D10 as the primary antibody at 24 hpi.</p>
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<p>Lymphocyte overexpression LAG3 promotes PRRSV replication in the co-culture system. (<b>A</b>) The expression levels of LAG3 expression at 24 and 48 hpt were detected by Western blotting. LAG3 mRNA expression (<b>B</b>) and PRRSV N gene copy number (<b>C</b>) in the co-culture system were quantified by RT-qPCR. The results are shown as mean ± SD from three independent experiments. NS means no significant difference, * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001. (<b>D</b>) LAG3 and PRRSV N protein expression in the co-culture system were detected by Western blotting. (<b>E</b>) The detection result of virus titer in the co-culture supernatant.</p>
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<p>PRRSV replication was inhibited when interference LAG3 expression in lymphocytes with siRNA. The interference effect of specific siRNA was detected by (<b>A</b>) RT-qPCR and (<b>B</b>) Western blotting. The results are shown as mean ± SD from three independent experiments. NS means no significant difference, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01. (<b>C</b>) LAG3 gene expression and (<b>D</b>) PRRSV N gene copy number in the co-culture system were analyzed by RT-qPCR. (<b>E</b>) LAG3 and PRRSV N protein expression in the co-culture system were analyzed by Western blotting. (<b>F</b>) The detection result of virus titer in the co-culture supernatant.</p>
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<p>mAb 1C2 blocking inhibited PRRSV replication in the co-culture system. (<b>A</b>) RT-qPCR detection results of PRRSV N gene copy number in the co-culture system. The results are shown as mean ± SD from three independent experiments. * <span class="html-italic">p</span> &lt; 0.05. (<b>B</b>) Western-blotting results of PRRSV N protein expression in the co-culture system. (<b>C</b>) The detection result of virus titer in the co-culture supernatant.</p>
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5 pages, 337 KiB  
Communication
The Influence of Specific Pathogen-Free and Conventional Environments on the Hematological Parameters of Pigs Bred for Xenotransplantation
by Won Kil Lee, Hwi-Cheul Lee, Seunghoon Lee, Haesun Lee, Sang Eun Kim, Minguk Lee, Jin-Gu No, Keon Bong Oh and Poongyeon Lee
Life 2024, 14(9), 1132; https://doi.org/10.3390/life14091132 - 8 Sep 2024
Viewed by 883
Abstract
Blood analysis plays a pivotal role in assessing the health of laboratory animals, including pigs. This study investigated the hematological profiles of transgenic pigs of the MGH breed for xenotransplantation, focusing on the effect of housing conditions on blood parameters. A cohort of [...] Read more.
Blood analysis plays a pivotal role in assessing the health of laboratory animals, including pigs. This study investigated the hematological profiles of transgenic pigs of the MGH breed for xenotransplantation, focusing on the effect of housing conditions on blood parameters. A cohort of pigs was longitudinally monitored from 6 to 18 months of age in both conventional and specific pathogen-free (SPF) environments. Red blood cells (RBCs), hemoglobin (HGB), and white blood cells (WBCs) were analyzed using standardized hematology analyzers. The results revealed that RBC and HGB levels were consistently higher in SPF-housed pigs. Notably, WBC counts were significantly lower in SPF-housed pigs, suggesting that reduced pathogen exposure under SPF conditions effectively diminished immune system activation. These findings raise a novel question as to whether distinct hematological parameters of specific and/or designated PF pigs would be advantages for the success of clinical xenotransplantation trials. Full article
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<p>Hematological profiles of GTKO/MCP transgenic pigs over time in different housing environments. Panels show levels of red blood cells (RBC, Panel (<b>A</b>)), hemoglobin (HGB, Panel (<b>B</b>)), and white blood cells (WBC, Panel (<b>C</b>)) at 6, 12, and 18 months in conventional (Con) and specific pathogen-free (SPF) facilities. The values are presented as mean ± SD, with sample sizes at each time point indicated as follows: at 6 months (Con: n = 58, SPF: n = 13), at 12 months (Con: n = 52, SPF: n = 7), and at 18 months (Con: 49, SPF: n = 5). These numbers reflect the volume of pigs analyzed in each housing condition at every specified age point. Asterisks indicate statistical significance compared to the conventional facility at the same time point (* <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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17 pages, 1961 KiB  
Article
Knockdown of Gonadotropin-Releasing Hormone II Receptor Impairs Ovulation Rate, Corpus Luteum Development, and Progesterone Production in Gilts
by Amy T. Desaulniers, Rebecca A. Cederberg, Clay A. Lents and Brett R. White
Animals 2024, 14(16), 2350; https://doi.org/10.3390/ani14162350 - 14 Aug 2024
Viewed by 1107
Abstract
Reproduction is classically controlled by gonadotropin-releasing hormone (GnRH-I) and its receptor (GnRHR-I) within the brain. In pigs, a second form (GnRH-II) and its specific receptor (GnRHR-II) are also produced, with greater abundance in peripheral vs. central reproductive tissues. The binding of GnRH-II to [...] Read more.
Reproduction is classically controlled by gonadotropin-releasing hormone (GnRH-I) and its receptor (GnRHR-I) within the brain. In pigs, a second form (GnRH-II) and its specific receptor (GnRHR-II) are also produced, with greater abundance in peripheral vs. central reproductive tissues. The binding of GnRH-II to GnRHR-II has been implicated in the autocrine/paracrine regulation of gonadal steroidogenesis rather than gonadotropin secretion. Blood samples were collected from transgenic gilts, with the ubiquitous knockdown of GnRHR-II (GnRHR-II KD; n = 8) and littermate controls (n = 7) at the onset of estrus (follicular) and 10 days later (luteal); serum concentrations of 16 steroid hormones were quantified by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Upon euthanasia, ovarian weight (OWT), ovulation rate (OR), and the weight of each excised Corpus luteum (CLWT) were recorded; HPLC-MS/MS was performed on CL homogenates. During the luteal phase, serum progesterone concentration was reduced by 18% in GnRHR-II KD versus control gilts (p = 0.0329). Age and weight at puberty, estrous cycle length, and OWT were similar between lines (p > 0.05). Interestingly, OR was reduced (p = 0.0123), and total CLWT tended to be reduced (p = 0.0958) in GnRHR-II KD compared with control females. Luteal cells in CL sections from GnRHR-II KD gilts were hypotrophic (p < 0.0001). Therefore, GnRH-II and its receptor may help regulate OR, CL development, and progesterone production in gilts. Full article
(This article belongs to the Special Issue Endocrinology of the Female Reproductive System)
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<p>Experimental design. The once daily detection of estrus began at approximately 170 d of age. Puberty was considered the first display of behavioral estrus. The detection of estrus continued for a total of five consecutive estrous cycles. At the onset of the third estrous cycle (d 0), blood was collected via jugular venipuncture (follicular sample) and 10 d later (luteal sample). Animals were euthanized and reproductive tissues were collected on approximately day 7 of the fifth estrous cycle.</p>
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<p>Body weights were not different between GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts over time. Body weight was recorded at birth, weaning, and during pre-pubertal development (40, 60, 80, 100, 125, 145 and 165 d of age). Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). Line, <span class="html-italic">p</span> = 0.3677; Age, <span class="html-italic">p</span> &lt; 0.0001; Line × Age, <span class="html-italic">p</span> = 0.7960.</p>
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<p>Age at puberty (<b>a</b>), weight at puberty (<b>b</b>), and estrous cycle length (<b>c</b>) in GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts. No line effects were detected (<span class="html-italic">p</span> &gt; 0.10). Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM).</p>
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<p>Concentrations of 11-deoxycortisol in blood serum samples from GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts during the follicular and luteal phases of the estrous cycle. There was no effect of phase or line by phase interaction (<span class="html-italic">p</span> &gt; 0.05). However, there was an overall effect (<span class="html-italic">p</span> = 0.0320) of line; GnRHR-II KD gilts had reduced circulating concentrations. Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Progestogen concentrations in blood serum samples from GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts during the follicular and luteal phase of the estrous cycle. Neither an effect of line (GnRHR-II KD versus control) nor a line by phase interaction was detected for 17α-hydroxyprogesterone, so these data are not reported. However, a phase effect (<span class="html-italic">p</span> = 0.0006) was detected for 17α-hydroxyprogesterone, with the concentration greater during the luteal phase (<b>a</b>). A line by phase interaction (<span class="html-italic">p</span> = 0.0341) was detected for progesterone; GnRHR-II KD gilts produced less progesterone during the luteal phase (<b>b</b>). Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). <sup>a,b,c</sup> Divergent letters differ significantly (<span class="html-italic">p</span> &lt; 0.05); * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Androgen concentrations during the follicular phase and luteal phase in blood serum samples from GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts. No effect of line (GnRHR-II KD versus control) nor line by phase interaction was detected for any androgen examined (<span class="html-italic">p</span> &gt; 0.05); therefore, these data are not reported. A phase effect (<span class="html-italic">p</span> &lt; 0.05) was detected for testosterone (<b>a</b>), androsterone (<b>b</b>) and androstenedione (<b>c</b>) with concentrations greater during the follicular phase. Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Estrogen concentrations during the follicular and luteal phase in blood serum samples from GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts. No effect of line (GnRHR-II KD versus control) nor line by phase interaction was detected for estrogens (<span class="html-italic">p</span> &gt; 0.05); therefore, these data are not reported. A phase effect (<span class="html-italic">p</span> &lt; 0.05) was detected for both 17β-estradiol (<b>a</b>) and estrone (<b>b</b>) with concentrations greater during the follicular phase compared with luteal phase. Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Ovarian characteristics in GnRHR-II KD (<span class="html-italic">n</span> = 8) and littermate control (<span class="html-italic">n</span> = 7) gilts. Paired ovary weight (<b>a</b>) was similar (<span class="html-italic">p</span> &gt; 0.10) between lines, but ovulation rate (number of <span class="html-italic">Corpora lutea</span>) (<b>b</b>) was reduced (<span class="html-italic">p</span> = 0.0123) in GnRHR-II KD gilts compared with littermate controls. Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p><span class="html-italic">Corpus luteum</span> (CL) metrics and GnRHR-II expression in CL samples from GnRHR-II KD and littermate control gilts. Average individual CL weight was greater (<span class="html-italic">p</span> &lt; 0.0001) in GnRHR-II KD (<span class="html-italic">n</span> = 4) versus control (<span class="html-italic">n</span> = 4) gilts (<b>a</b>). Total CL weight per ovary tended to be reduced (<span class="html-italic">p</span> = 0.0958) in GnRHR-II KD (<span class="html-italic">n</span> = 4) versus control (<span class="html-italic">n</span> = 4) gilts (<b>b</b>). Luteal cell area was reduced (<span class="html-italic">p</span> &lt; 0.0001) in GnRHR-II KD (<span class="html-italic">n</span> = 7) versus littermate control (<span class="html-italic">n</span> = 6) gilts (<b>c</b>). Expression of GnRHR-II tended to be reduced (<span class="html-italic">p</span> = 0.0774) by 21% in CL from GnRHR-II KD (<span class="html-italic">n</span> = 8) versus control (<span class="html-italic">n</span> = 7) gilts (<b>d</b>). Results are presented as least squares means (LSMEANS) ± the standard error of the mean (SEM). * <span class="html-italic">p</span> &lt; 0.05; † <span class="html-italic">p</span> &lt; 0.10.</p>
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9 pages, 240 KiB  
Article
Exploring Serum Copeptin and Hematological Profile: A Comparative Analysis after Intradermal versus Intramuscular Porcine Reproductive and Respiratory Syndrome Virus Vaccination in Piglets
by Georgios Maragkakis, Eleni G. Katsogiannou, Georgios I. Papakonstantinou, Laskarina-Maria Korou, Serafeim C. Chaintoutis, Panagiotis Konstantopoulos, Despoina N. Perrea, Georgios Christodoulopoulos, Labrini V. Athanasiou and Vasileios G. Papatsiros
Stresses 2024, 4(2), 358-366; https://doi.org/10.3390/stresses4020023 - 5 Jun 2024
Viewed by 1228
Abstract
This study aimed to investigate the impact of intradermal (ID) and intramuscular (IM) vaccination with a porcine reproductive and respiratory syndrome virus (PRRSV)-modified live vaccine (MLV) in piglets on serum copeptin levels and hematological profile. This study included 104 suckling piglets (2 weeks [...] Read more.
This study aimed to investigate the impact of intradermal (ID) and intramuscular (IM) vaccination with a porcine reproductive and respiratory syndrome virus (PRRSV)-modified live vaccine (MLV) in piglets on serum copeptin levels and hematological profile. This study included 104 suckling piglets (2 weeks of age) from a commercial farrow-to-finish pig farm suffering from positive unstable PRRSV status. Animals were assigned to four groups, with two replicates (13 piglets/group/replicate); group A: IM vaccination with a PRRSV MLV vaccine, group B: ID vaccination with the same vaccine, group C: ID of Diluvac Forte, and group D: IM of Diluvac Forte. Blood samples were collected from the same three pigs/group/replicate at 4, 7, and 10 weeks of age. Blood samples were used for the performance of the complete blood count, and they were also examined by PCR for PRRSV and by ELISA for copeptin. No significant differences in serum copeptin levels and the number of blood cell counts (packed cell volume—PCV, numbers of white blood cells—WBCs, and platelets number—PLTs) were noticed in the same group over time and among groups. In conclusion, it seems that the vaccination against PRRSV does not affect the levels of the released copeptin. Based on our results, the measurement of serum copeptin could not be proposed as a potential stress biomarker in pigs. Full article
(This article belongs to the Collection Feature Papers in Human and Animal Stresses)
8 pages, 1920 KiB  
Article
Characterization of Japanese Encephalitis Virus Isolated from Persistently Infected Mouse Embryo Cells
by Yume Kondo and Tomoyoshi Komiya
Trop. Med. Infect. Dis. 2024, 9(5), 117; https://doi.org/10.3390/tropicalmed9050117 - 16 May 2024
Viewed by 1374
Abstract
Japanese encephalitis virus (JEV) has a positive-sense single-stranded RNA genome and belongs to the genus Flavivirus of the family Flaviviridae. Persistent JEV infection was previously shown in pig blood cells, which act as a natural reservoir of this virus. We aimed to [...] Read more.
Japanese encephalitis virus (JEV) has a positive-sense single-stranded RNA genome and belongs to the genus Flavivirus of the family Flaviviridae. Persistent JEV infection was previously shown in pig blood cells, which act as a natural reservoir of this virus. We aimed to determine the pathogenicity factors involved in persistent JEV infection by analyzing the pathogenicity and genome sequences of a virus isolated from a persistent infection model. We established persistent JEV infections in cells by inoculating mouse fetus primary cell cultures with the Beijing-1 strain of JEV and then performing repeated infected cell passages, harvesting viruses after each passage while monitoring the plaque size over 100 generations. The virus growth rate was compared among Vero, C6/36, and Neuro-2a cells. The pathogenicity was examined in female ICR mice at several ages. Additionally, we determined the whole-genome sequences. The 134th Beijing-1-derived persistent virus (ME134) grew in Vero cells at a similar rate to the parent strain but did not grow well in C6/36 or Neuro-2a cells. No differences were observed in pathogenicity after intracerebral inoculation in mice of different ages, but the survival time was extended in older mice. Mutations in the persistent virus genomes were found across all regions but were mainly focused in the NS3, NS4b, and 3′NCR regions, with a 34-base-pair deletion found in the variable region. The short deletion in the 3′NCR region appeared to be responsible for the reduced pathogenicity and growth efficiency. Full article
(This article belongs to the Special Issue Japanese Encephalitis)
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<p>Plaque sizes of the persistent and parent viruses. The ME134 persistent strain virus (<b>A</b>) and the Beijing-1 strain virus (<b>B</b>) were titrated on Vero cells.</p>
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<p>Immunofluorescence assay of ME cells using anti-JEV NS1 antibody after one day of subculture. NS1 protein (<b>A</b>) and nuclei (<b>B</b>) were detected using the anti-JEV NS1 antibody and DAPI, respectively, in mouse embryo cells infected with ME virus.</p>
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<p>Virus growth rates for the persistent and parent virus strains. Vero cells (<b>A</b>), SK-N-SH cells (<b>B</b>), Neuro-2a cells (<b>C</b>), and C6/36 cells (<b>D</b>) were infected with the viruses at MOI = 0.01 and cultured for 6 days.</p>
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<p>Survival rate after infection with 10<sup>4</sup> or 10<sup>5</sup> of each virus in 4-week-old ICR mice (<b>A</b>) and 2-day-old ICR mice (<b>B</b>), respectively, inoculated via the i.c. route.</p>
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<p>Sequence analysis in the 3′ noncoding region. The SA-14-14-2 strain is a live attenuated vaccine strain, and the Ishikawa strain is a genotype I strain isolated in Japan.</p>
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14 pages, 1718 KiB  
Article
Preweaning Purified Fucoidan Drench: Effects on Growth, Immune Response, and Intestinal Morphology in Weaned Piglets
by Bonnie Homer, Reza Barekatain, Kiro R. Petrovski, Kate J. Plush, Corinna Dwan, Darryl N. D’Souza, Paul J. Verma, Roy N. Kirkwood and Bryony S. Tucker
Animals 2024, 14(10), 1472; https://doi.org/10.3390/ani14101472 - 15 May 2024
Cited by 1 | Viewed by 1165
Abstract
Weaning stress imposes considerable physiological challenges on piglets, often manifesting in intestinal disturbances, such as inflammation and compromised barrier function, ultimately affecting growth and health outcomes. While conventional interventions, including antimicrobials, have effectively mitigated these sequelae, concerns surrounding antimicrobial resistance necessitate the exploration [...] Read more.
Weaning stress imposes considerable physiological challenges on piglets, often manifesting in intestinal disturbances, such as inflammation and compromised barrier function, ultimately affecting growth and health outcomes. While conventional interventions, including antimicrobials, have effectively mitigated these sequelae, concerns surrounding antimicrobial resistance necessitate the exploration of alternatives. Fucoidan, derived from brown seaweed, offers promise due to its antioxidant and anti-inflammatory effects. Previous research has been limited to the in-feed supplementation of partially purified fucoidan extracted from brown seaweed. The focus of the present study is assessing the effect of a preweaning drench with highly purified (85%) fucoidan on piglet growth, immune response, and intestinal morphology post-weaning. Forty-eight male piglets at 17 ± 3 days of age (5.67 ± 0.16 kg) were assigned to a saline (control), fucoidan, or antimicrobial group, receiving treatment as a single 18 mL oral drench three days before weaning. Monitoring for seven days post-weaning included body weight measurements, blood sample collection for the inflammatory protein assay, and small intestine morphological analysis. The findings revealed that the preweaning fucoidan drench did not elicit adverse effects on piglets. However, neither fucoidan nor antimicrobial drenches significantly enhanced growth parameters, immune markers, or intestinal morphology compared to that of the control-treated piglets (p > 0.05). The lack of response may be attributed to the high health status of the experimental cohort and the limitation of a single dosage. Future research should consider a more challenging production setting to evaluate the viability and optimal application of fucoidan as an antimicrobial alternative in the pig industry. Full article
(This article belongs to the Special Issue Piglets Nutrition and Management)
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<p>The 28-day study timeline with observational and intervention phases. The experimental day represents the average age of the intervention phase animals within a range of ±3 days.</p>
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<p>Diagram illustrating the conventional farrowing crate layout where piglets were housed with their birth sows until weaning.</p>
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<p>Schematic representation of the weaner pen layout where piglets were housed in their treatment groups (<span class="html-italic">n</span> = 16 piglets/pen) from weaning until the end of the trial.</p>
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<p>Histology image (500 µm) depicting morphometric measurements for the villous height (straight line) and the crypt depth (oval) from a jejunal tissue section (haematoxylin and eosin stained) of a piglet euthanized eight days post-weaning.</p>
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13 pages, 260 KiB  
Article
The Effects of Microbial Additive Supplementation on Growth Performance, Blood Metabolites, Fecal Microflora, and Carcass Characteristics of Growing–Finishing Pigs
by Hyuk-Jun Lee, Bu-Gil Choi, Young-Ho Joo, Chang-Hyun Baeg, Ji-Yoon Kim, Dong-Hyeon Kim, Seong-Shin Lee and Sam-Churl Kim
Animals 2024, 14(9), 1268; https://doi.org/10.3390/ani14091268 - 23 Apr 2024
Viewed by 1685
Abstract
This study aimed to assess the effects of microbial additives that produce antimicrobial and digestive enzymes on the growth performance, blood metabolites, fecal microflora, and carcass characteristics of growing–finishing pigs. A total of 180 growing–finishing pigs (Landrace × Yorkshire × Duroc; mixed sex; [...] Read more.
This study aimed to assess the effects of microbial additives that produce antimicrobial and digestive enzymes on the growth performance, blood metabolites, fecal microflora, and carcass characteristics of growing–finishing pigs. A total of 180 growing–finishing pigs (Landrace × Yorkshire × Duroc; mixed sex; 14 weeks of age; 58.0 ± 1.00 kg) were then assigned to one of three groups with three repetitions (20 pigs) per treatment for 60 days of adaptation and 7 days of collection. Dietary treatments included 0, 0.5, and 1.0% microbial additives in the basal diet. For growth performance, no significant differences in the initial and final weights were observed among the dietary microbial additive treatments, except for the average daily feed intake, average daily gain, and feed efficiency. In terms of blood metabolites and fecal microflora, immunoglobulin G (IgG), blood urea nitrogen, blood glucose, and fecal lactic acid bacteria count increased linearly, and fecal E. coli counts decreased linearly with increasing levels of microbial additives but not growth hormones and Salmonella. Carcass quality grade was improved by the microbial additive. In addition, carcass characteristics were not influenced by dietary microbial additives. In conclusion, dietary supplementation with 1.0% microbial additive improved average daily gain, feed efficiency, IgG content, and fecal microflora in growing–finishing pigs. Full article
(This article belongs to the Special Issue Feed Additives in Pig Feeding: 2nd Edition)
14 pages, 8091 KiB  
Article
Integrated Microbiome and Serum Metabolome Analysis Reveals Molecular Regulatory Mechanisms of the Average Daily Weight Gain of Yorkshire Pigs
by Shan Jiang, Jinglei Si, Jiayuan Mo, Shuai Zhang, Kuirong Chen, Jiuyu Gao, Di Xu, Lijing Bai, Ganqiu Lan and Jing Liang
Animals 2024, 14(2), 278; https://doi.org/10.3390/ani14020278 - 16 Jan 2024
Viewed by 1868
Abstract
The average daily weight gain (ADG) is considered a crucial indicator for assessing growth rates in the swine industry. Therefore, investigating the gastrointestinal microbiota and serum metabolites influencing the ADG in pigs is pivotal for swine breed selection. This study involved the inclusion [...] Read more.
The average daily weight gain (ADG) is considered a crucial indicator for assessing growth rates in the swine industry. Therefore, investigating the gastrointestinal microbiota and serum metabolites influencing the ADG in pigs is pivotal for swine breed selection. This study involved the inclusion of 350 purebred Yorkshire pigs (age: 90 ± 2 days; body weight: 41.20 ± 4.60 kg). Concurrently, serum and fecal samples were collected during initial measurements of blood and serum indices. The pigs were categorized based on their ADG, with 27 male pigs divided into high-ADG (HADG) and low-ADG (LADG) groups based on their phenotype values. There were 12 pigs in LADG and 15 pigs in HADG. Feces and serum samples were collected on the 90th day. Microbiome and non-targeted metabolomics analyses were conducted using 16S rRNA sequencing and liquid chromatography-mass spectrometry (LC-MS). Pearson correlation, with Benjamini–Hochberg (BH) adjustment, was employed to assess the associations between these variables. The abundance of Lactobacillus and Prevotella in LADG was significantly higher than in HADG, while Erysipelothrix, Streptomyces, Dubosiella, Parolsenella, and Adlercreutzia in LADG were significantly lower than in HADG. The concentration of glutamine, etiocholanolone glucuronide, and retinoyl beta-glucuronide in LADG was significantly higher than in HADG, while arachidonic acid, allocholic acid, oleic acid, phenylalanine, and methyltestosterone in LADG were significantly lower than in HADG. The LactobacillusStreptomyces networks (Lactobacillus, Streptomyces, methyltestosterone, phenylalanine, oleic acid, arachidonic acid, glutamine, 3-ketosphingosine, L-octanoylcarnitine, camylofin, 4-guanidinobutyrate 3-methylcyclopentadecanone) were identified as the most influential at regulating swine weight gain. These findings suggest that the gastrointestinal tract regulates the daily weight gain of pigs through the network of Lactobacillus and Streptomyces. However, this study was limited to fecal and serum samples from growing and fattening boars. A comprehensive consideration of factors affecting the daily weight gain in pig production, including gender, parity, season, and breed, is warranted. Full article
(This article belongs to the Special Issue Molecular Mechanisms Affecting Important Traits of Pigs)
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<p>The α-diversity, β-diversity, and taxonomy between HADG and LADG. (<b>A</b>) The chao1 index. (<b>B</b>) The Shannon index. (<b>C</b>) The ACE index. (<b>D</b>) β-diversity. (<b>E</b>) The distribution of microbiota at the phylum level. (<b>F</b>) The distribution of microbiota at the genus level. If the small letters are the same, it indicates no significant difference (<span class="html-italic">p</span> &gt; 0.05).</p>
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<p>The Stamp analysis and BugBase analysis between HADG and LADG. (<b>A</b>) Microbiota with significantly different abundance at the genus level. (<b>B</b>) Contribution rate in the formation of biofilm. (<b>C</b>) Contribution rate in potentially pathogenic organisms. (<b>D</b>) Contribution rate in aerobics. NS: No Significance, *: <span class="html-italic">p</span> &lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>The PCA, OPLS-DA, and permutation score plots of serum samples collected from LADG and HADG based on LC-MS. (<b>A</b>) PCA score plot in the positive ion mode. (<b>B</b>) PCA score plot in the negative ion mode. (<b>C</b>) OPLS-DA score plot in the positive ion mode. (<b>D</b>) OPLS-DA score plot in the negative ion mode. (<b>E</b>) Permutation score plot in the positive ion mode. (<b>F</b>) Permutation score plot in the negative ion mode.</p>
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<p>The heatmap of differential metabolites between LADG and HADG. The labels “P” and “N” denote positive and negative ion modes, respectively.</p>
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<p>The KEGG pathways displayed enrichment in differential metabolites between LADG and HADG.</p>
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<p>The <span class="html-italic">co-occurrence</span> network of fecal microbiota and serum metabolites.</p>
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16 pages, 1904 KiB  
Article
Relation of Plasma Catecholamine Concentrations and Myocardial Mitochondrial Respiratory Activity in Anesthetized and Mechanically Ventilated, Cardiovascular Healthy Swine
by Nadja Abele, Franziska Münz, Fabian Zink, Michael Gröger, Andrea Hoffmann, Eva-Maria Wolfschmitt, Melanie Hogg, Enrico Calzia, Christiane Waller, Peter Radermacher and Tamara Merz
Int. J. Mol. Sci. 2023, 24(24), 17293; https://doi.org/10.3390/ijms242417293 - 9 Dec 2023
Cited by 1 | Viewed by 1357
Abstract
Chronic heart failure is associated with reduced myocardial β-adrenergic receptor expression and mitochondrial function. Since these data coincide with increased plasma catecholamine levels, we investigated the relation between myocardial β-receptor expression and mitochondrial respiratory activity under conditions of physiological catecholamine concentrations. This post [...] Read more.
Chronic heart failure is associated with reduced myocardial β-adrenergic receptor expression and mitochondrial function. Since these data coincide with increased plasma catecholamine levels, we investigated the relation between myocardial β-receptor expression and mitochondrial respiratory activity under conditions of physiological catecholamine concentrations. This post hoc analysis used material of a prospective randomized, controlled study on 12 sexually mature (age 20–24 weeks) Early Life Stress or control pigs (weaning at day 21 and 28–35 after birth, respectively) of either sex. Measurements in anesthetized, mechanically ventilated, and instrumented animals comprised serum catecholamine (liquid-chromatography/tandem-mass-spectrometry) and 8-isoprostane levels, whole blood superoxide anion concentrations (electron spin resonance), oxidative DNA strand breaks (tail moment in the “comet assay”), post mortem cardiac tissue mitochondrial respiration, and immunohistochemistry (β2-adrenoreceptor, mitochondrial respiration complex, and nitrotyrosine expression). Catecholamine concentrations were inversely related to myocardial mitochondrial respiratory activity and β2-adrenoceptor expression, whereas there was no relation to mitochondrial respiratory complex expression. Except for a significant, direct, non-linear relation between DNA damage and noradrenaline levels, catecholamine concentrations were unrelated to markers of oxidative stress. The present study suggests that physiological variations of the plasma catecholamine concentrations, e.g., due to physical and/or psychological stress, may affect cardiac β2-adrenoceptor expression and mitochondrial respiration. Full article
(This article belongs to the Special Issue New Cardiovascular Risk Factors)
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<p>Immunohistochemistry examples of the cardiac tissue protein expression of the (<b>a</b>) mitochondrial NADH dehydrogenase (ubiquinone oxidoreductase, complex I); (<b>b</b>) succinate dehydrogenase (complex II); (<b>c</b>) β<sub>2</sub>-adrenergic receptor, and (<b>d</b>) nitrotyrosine formation. Note that no nitrotyrosine staining could be detected, indicating that cardiac tissue oxidative and nitrosative stress was negligible if present at all. All pictures are displayed with a magnification of 10×.</p>
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<p>(<b>a</b>) Myocardial tissue maximum respiratory capacity in the coupled state (OxPhos) plotted as a function of the noradrenaline concentration; (<b>b</b>) myocardial tissue OxPhos plotted as a function of the adrenaline concentration; (<b>c</b>) myocardial tissue maximum electron transfer capacity in the uncoupled state (ETC) plotted as a function of the noradrenaline concentration, and (<b>d</b>) myocardial tissue ETC plotted as a function of the adrenaline concentration. Males are presented by closed squares, females by open circles, presence or absence of early life stress (ELS) is depicted by red and blue symbols, respectively. While OxPhos and ETC showed a significant, inverse linear correlation with the noradrenaline levels (r = −0.68, <span class="html-italic">p</span> = 0.015, and r = −0.76, <span class="html-italic">p</span> = 0.004, respectively), a significant, inverse non-linear correlation was present for ETC as a function of the adrenaline concentrations (r = −0.69, <span class="html-italic">p</span> = 0.013). The inverse non-linear relation of OxPhos plotted as a function of the adrenaline levels narrowly missed statistical significance (r = −0.57, <span class="html-italic">p</span> = 0.051).</p>
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<p>(<b>a</b>) Whole blood superoxide anion (O<sub>2</sub><sup>•−</sup>) concentrations; (<b>b</b>) whole blood tail moment in the comet assay, and (<b>c</b>) plasma isoprostane levels plotted as function of the noradrenaline concentration, and (<b>d</b>–<b>f</b>) the same parameters of oxidative stress plotted as a function of the adrenaline concentration. Males are presented by closed squares, females by open circles, presence or absence of ELS is depicted by red and blue symbols, respectively. Except for a significant, direct, non-linear correlation between the tail moment and the noradrenaline levels (r = 0.67, <span class="html-italic">p</span> = 0.017), no significant relation was detected.</p>
Full article ">Figure 4
<p>Results of the immunohistochemistry quantification of the myocardial tissue expression of the (<b>a</b>) mitochondrial respiratory complex I, (<b>b</b>) mitochondrial respiratory complex II, and (<b>c</b>) β<sub>2</sub>-adrenergic receptor plotted as a function of the noradrenaline; and (<b>d</b>–<b>f</b>) the same parameters plotted as a function of the adrenaline concentrations. Males are presented by closed squares, females by open circles, presence or absence of ELS is depicted by red and blue symbols, respectively. While there was no significant relation between the expression of the mitochondrial respiratory complexes and the catecholamine levels, the expression of the β<sub>2</sub>-adrenergic receptor showed a significant, inverse, linear correlation with both the noradrenaline (r = −0.59, <span class="html-italic">p</span> = 0.045) and adrenaline (r = −0.81, <span class="html-italic">p</span> = 0.001) concentrations.</p>
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16 pages, 294 KiB  
Article
Sustenance Trial to Analyze the Effects of Black Soldier Fly Larvae Meal on the Reproductive Efficiency of Sows and the Hematological Properties of Suckling and Weaning Piglets
by Kiyonori Kawasaki, Junliang Zhao, Natsu Takao, Masaki Sato, Takuma Ban, Kaoru Tamamaki, Masanori Kagami and Kiminobu Yano
Animals 2023, 13(21), 3410; https://doi.org/10.3390/ani13213410 - 3 Nov 2023
Cited by 2 | Viewed by 1680
Abstract
The escalating demand for meat, driven by global population growth, necessitates sustainable solutions for animal feed production. This study investigated the effects of substituting conventional protein resources in sow and piglet dietary regimens with black soldier fly (BSF; Hermetia illucens) meal on [...] Read more.
The escalating demand for meat, driven by global population growth, necessitates sustainable solutions for animal feed production. This study investigated the effects of substituting conventional protein resources in sow and piglet dietary regimens with black soldier fly (BSF; Hermetia illucens) meal on reproductive efficiency, blood profile, piglet growth, and intestinal tissue morphology. The results indicate that substituting animal-derived and soy proteins with BSF meal does not compromise sow reproductive performance. Although no notable disparities were observed in piglet growth, the feed conversion ratio from the 28- to 35-day age marks were lower in the BSF-fed groups. This suggests that the animal protein-BSF substitution rate may require optimization, potentially involving chitin removal from BSF meal to enhance digestibility. Minor variations in the hematological composition and properties in piglets, with elevated high-density lipoprotein cholesterol levels in the high BSF group at the 28-day mark, were potentially attributable to the unique fatty acid composition of BSF meal. Moreover, this study potentiates future exploration into the efficacy of complete animal protein substitution with BSF meals on piglet nutrition and physiology, particularly in fattening pigs. The practical implementation of BSF meals in animal feed production holds promise for enhancing the sustainability of the swine industry. Full article
(This article belongs to the Collection Insects as Animal Feed: A New Promising Sector)
18 pages, 385 KiB  
Review
Telomere Dynamics in Livestock
by Nan Zhang, Emilie C. Baker, Thomas H. Welsh and David G. Riley
Biology 2023, 12(11), 1389; https://doi.org/10.3390/biology12111389 - 31 Oct 2023
Cited by 2 | Viewed by 2737
Abstract
Telomeres are repeated sequences of nucleotides at the end of chromosomes. They deteriorate across mitotic divisions of a cell. In Homo sapiens this process of lifetime reduction has been shown to correspond with aspects of organismal aging and exposure to stress or other [...] Read more.
Telomeres are repeated sequences of nucleotides at the end of chromosomes. They deteriorate across mitotic divisions of a cell. In Homo sapiens this process of lifetime reduction has been shown to correspond with aspects of organismal aging and exposure to stress or other insults. The early impetus to characterize telomere dynamics in livestock related to the concern that aged donor DNA would result in earlier cell senescence and overall aging in cloned animals. Telomere length investigations in dairy cows included breed effects, estimates of additive genetic control (heritability 0.12 to 0.46), and effects of external stressors on telomere degradation across animal life. Evaluation of telomeres with respect to aging has also been conducted in pigs and horses, and there are fewer reports of telomere biology in beef cattle, sheep, and goats. There were minimal associations of telomere length with animal productivity measures. Most, but not all, work in livestock has documented an inverse relationship between peripheral blood cell telomere length and age; that is, a longer telomere length was associated with younger age. Because livestock longevity affects productivity and profitability, the role of tissue-specific telomere attrition in aging may present alternative improvement strategies for genetic improvement while also providing translational biomedical knowledge. Full article
(This article belongs to the Section Biotechnology)
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