Pig Spleen Research Articles

Deoxynivalenol Induces Local Inflammation and Lesions in Tissues at Doses Recommended by the EU.

The mycotoxin deoxynivalenol (DON) is frequently present in cereals at low levels, resulting in its occurrence in food and feed. DON has been proven to alter the immune response and induce inflammation in all species, with pigs exhibiting heightened sensitivity and exposure. However, no study has yet evaluated the effects of exposure to DON at the recommended levels in pig feed. In two separate trials, piglets were subjected to control feed or feed contaminated with a low level of purified DON (0.83 mg/kg feed in trial 1 and 0.85 mg/kg feed in trial 2) for either three weeks (trial 1) or two weeks (trial 2). Additionally, a group of animals exposed to 2.85 mg/kg feed of DON was included as a positive control in Trial 1. The impact of DON on porcine tissues (intestine, liver, and spleen) was evaluated through histological and qPCR analyses of immune-related genes. Additionally, biochemical analyses and acute-phase proteins were examined in plasma samples. Lesions were identified in the intestine (jejunum and ileum), the liver, and the spleen of pigs receiving diets contaminated with low and high concentrations of DON. The low level of DON also resulted in impaired expression of genes associated with intestinal barrier integrity, intestinal immune responses, and liver function. In conclusion, the results of the two trials demonstrate the impact of DON exposure even at doses below the recommended level of 0.9 mg/kg feed set by the European Union. This suggests that the current recommended level should be reconsidered to ensure the optimal health and well-being of pigs.

The Comparative Full-Length Genome Characterization of African Swine Fever Virus Detected in Thailand.

African swine fever virus (ASFV) has been responsible for the globally devastating epidemics in wild and domesticated pigs. Of the 24 identified ASFV genotypes, genotype II is the primary cause for the pandemic occurring in Europe and Asia since its emergence in Georgia in 2007. The current study aimed to characterize the full-length genomic pattern of the ASFV strain from Thailand, TH1_22/CR (Accession No. PP915735), which was then compared with genomic diversity across other Asian isolates using Georgia 2007/1 (Accession No. FR682468) as the reference. Viral DNA was isolated from the pig spleen sample following library preparation and paired-end sequencing using the MiSeq Illumina platform. The sequenced TH1_22/CR isolate spanned 189,395 nucleotides encoding 193 open reading frames (ORFs), exhibiting maximum nucleotide similarity (99.99%) with Georgian (Georgia 2007/1) and Chinese (Wuhan 2019-1 and China HLJ) isolates. Based on phylogenetic analysis, the TH1_22/CR isolate (Accession No. PP915735) was characterized as genotype II, serogroup 8, and IGR-II due to the presence of three tandem repeat sequences (TRSs). Genetic variations including SNPs and single and polynucleotide indels were identified in TH1_22/CR in agreement with other Asian isolates. For comprehensive analysis, the genome was divided into four regions (I-IV) based on gene location. Overall, the TH1_22/CR isolate demonstrated eight SNPs and indels in its genome. Two unique SNPs were reported in the coding regions of the TH1_22/CR isolate, out of which, a C-591-T substitution was seen in MGF 360-4L and a C-297-T was found in A238L, and four unique SNPs were reported in non-coding regions (NCRs). Furthermore, a 29 bp deletion was observed in the IGR between MGF 110-13La and MGF 110-13Lb, as well as 52 bp deletion in the ASFV G ACD 00350 gene. This comparative analysis establishes the foundational information for future studies on the diversity and phylogeography of this regionally significant genetic sub-group of ASFV.

Reference Intervals of Mass Coefficients and Absolute Weights of Mini Pigs’ Organs

Introduction. Measurement of weights and calculation of mass coefficients of the laboratory animals’ internal organs are the integral stages of toxicological research. However, the absence of the correct intra-laboratory reference intervals corresponding to the normal values established for the animal population of a research laboratory could hinder the analysis of the mass coefficient changes in the experiment. For example, the data on weights and mass coefficients of mini pigs’ organs is being an issue due to its insufficient availability in the open sources. At the same time, in the literature there is a lot of such kind of data referring to the industrial pig breeds. Due to the wide use of the mini pigs in the pre-clinical research and because in the pig species taxonomy they are considered to be a weight category representing the pigs with reduced mass of the body with similar morphology of the organs and body systems, we have set an objective to determine and calculate the reference intervals of mass coefficients of internal organs relative to the mass of the body and brain, as well as the absolute weights of mini pigs’ internal organs and compare the results with the data on mini and industrial pig breeds in the literature sources.Materials and Methods. The data on 29 males and 29 females of the Vietnamese lop-bellied mini pig breed aged 6 months and older, who used to be in control groups of the various experiments held at JSC “SPA “House of Pharmacy” (Leningrad region) in the period from January 2018 to March 2024, was used to calculate the reference intervals. The statistical analysis was peRussian Federationormed using Microsoft Excel and Reference Value Advisor v2.1 software in compliance with the CLSI guidelines. Calculation of the reference intervals for all parameters was carried out by a robust method, the compliance with the normal distribution law was determined by Anderson–Darling test, in case of abnormal distribution of data, calculation of the reference interval was carried out with the Box–Cox transformation.Results. The data on the absolute weights of organs and their coefficients relative to the body and brain masses were presented in the form of the lower and upper reference limits and confidence intervals for each of the limit. The reference intervals of masses of organs across all parameters in males and females had similar limits, the most comparable values were found for the heart and brain parameters. The calculated interval of the heart absolute weight in males was 28.3–253 g, that of the brain was 46.5–93.7 g; in females these intervals were 42.9–279 g and 55.5–81.8 g respectively. Upon comparing the obtained data with the literature sources, it was found that the majority of the absolute weights of all measured mini pigs’ organs had fitted in the limits of the reference values, whereas the masses of the lungs and liver were at the lower limit or below the calculated intervals. The values of the absolute weights of most of the organs of the industrial pig breeds were higher than that of mini pigs, which was due to the difference in the body size and mass of the studied animal breeds. When comparing the mass coefficients of organs in relation to the body mass, it was found that the coefficients of most organs of the industrial breeds and mini pigs, on the contrary, were within the calculated reference intervals or slightly exceeded them.Discussion and conclusion. The calculated reference intervals of the mass coefficients of some organs in relation to the body mass, as well as the absolute weights of the spleen in mini pigs are comparable with the data available in literature for the industrial meat pig breeds. At the same time, the data available in the literature sources on the absolute weights of organs and their mass coefficients relative to the body mass of mini pigs, in most cases, have fitted into the calculated intervals. To increase the prognostic value of the obtained reference intervals, the analysis of a larger size sample is necessary.

A dicoumarol-graphene oxide quantum dot polymer inhibits porcine reproductive and respiratory syndrome virus through the JAK-STAT signaling pathway.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes substantial economic losses in the global swine industry. The current vaccine options offer limited protection against PRRSV transmission, and there are no effective commercial antivirals available. Therefore, there is an urgent need to develop new antiviral strategies that slow global PRRSV transmission. In this study, we synthesized a dicoumarol-graphene oxide quantum dot (DIC-GQD) polymer with excellent biocompatibility. This polymer was synthesized via an electrostatic adsorption method using the natural drug DIC and GQDs as raw materials. Our findings demonstrated that DIC exhibits high anti-PRRSV activity by inhibiting the PRRSV replication stage. The transcriptome sequencing analysis revealed that DIC treatment stimulates genes associated with the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signalling pathway. In porcine alveolar macrophages (PAMs), DIC-GQDs induce TYK2, JAK1, STAT1, and STAT2 phosphorylation, leading to the upregulation of JAK1, STAT1, STAT2, interferon-β (IFN-β) and interferon-stimulated genes (ISGs). Animal challenge experiments further confirmed that DIC-GQDs effectively alleviated clinical symptoms and pathological reactions in the lungs, spleen, and lymph nodes of PRRSV-infected pigs. These findings suggest that DIC-GQDs significantly inhibits PRRSV proliferation by activating the JAK/STAT signalling pathway. Therefore, DIC-GQDs hold promise as an alternative treatment for PRRSV infection.

Anatomic and Surgical Aspects of Total Pancreatectomy in Swine

IntroductionThe swine is valuable model for preclinical research and surgical technique training. From the induction of Diabetes through total pancreatectomy, these animals can be used in many research studies, such as islet transplantation. In view of the lack of elucidation in literature, the purpose of this work is to describe anatomical aspects, surgical technique of total pancreatectomy, intra- and postoperative complications and autopsy results Material and MethodFive male pigs, hybrids (Large White/Landrace), 20-35 kg, submitted to total pancreatectomy with duodenum, bile duct and spleen preservation. Postoperatively, daily clinical assessment and capillary blood glucose collection were performed. At the end of the 30-day limit or in the occurrence of serious clinical complications, euthanasia and autopsy were performed ResultsThe average duration of surgery was 128 minutes, without intraoperative deaths or anesthesia induction failures. Median survival was 6.6 days. Postoperative complications were weight loss (3), emesis (2), constipation (2), abdominal distension (2), diarrhea (1) and loss of appetite (1). All animals were euthanized due to serious complications. Two animals presented surgical complications (duodenal necrosis with gastroparesis and internal hernia with intestinal necrosis). The other three animals presented serious clinical complications related to exocrine pancreatic insufficiency due to deficiency of pancreatic enzymes. Glycemic values above 200 mg/dL were found on the first postoperative day and above 300 mg/dL on the seventh day in all animals Discussion and ConclusionsIn this experiment, a model of total pancreatectomy with duodenum, spleen and bile duct preservation in pigs was established. All animals had diabetes, however animals without postoperative complications were euthanized due to serious complications related to pancreas exocrine insufficiency.

Gross Morphometry of Spleen in Postnatal Development Period of Indian Domestic Pig (Sus scrofa domesticus)

The present postnatal developmental study was conducted on 18 healthy pigs (Sus Scrofa domesticus) irrespective of sex. The size, shape, colour, weight, volume, length, width, and thickness were studied. Gross features revealed that the spleen in the pig was an elongated flat lymphoid organ located on the left side of the abdominal cavity along the greater curvature of the stomach in close relation to it and the colon. The colour of the spleen changed from reddish brown in the early stages to dark brown with the advancement of age. The pig's spleen presented two surfaces, two borders and two extremities, of which the parietal surface was convex and related to the left lateral wall of the abdomen. The visceral surface was divided by a longitudinal ridge into the anterior area related to the stomach and the posterior area to the colon. Hilus was located on the ridge. All gross parameters increased with the advancement of age in this study.

Single-cell profiling of African swine fever virus disease in the pig spleen reveals viral and host dynamics

African swine fever, one of the major viral diseases of swine, poses an imminent threat to the global pig industry. The high-efficient replication of the causative agent African swine fever virus (ASFV) in various organs in pigs greatly contributes to the disease. However, how ASFV manipulates the cell population to drive high-efficient replication of the virus in vivo remains unclear. Here, we found that the spleen reveals the most severe pathological manifestation with the highest viral loads among various organs in pigs during ASFV infection. By using single-cell-RNA-sequencing technology and multiple methods, we determined that macrophages and monocytes are the major cell types infected by ASFV in the spleen, showing high viral-load heterogeneity. A rare subpopulation of immature monocytes represents the major population infected at late infection stage. ASFV causes massive death of macrophages, but shifts its infection into these monocytes which significantly arise after the infection. The apoptosis, interferon response, and antigen-presentation capacity are inhibited in these monocytes which benefits prolonged infection of ASFV in vivo. Until now, the role of immature monocytes as an important target by ASFV has been overlooked due to that they do not express classical monocyte marker CD14. The present study indicates that the shift of viral infection from macrophages to the immature monocytes is critical for maintaining prolonged ASFV infection in vivo. This study sheds light on ASFV tropism, replication, and infection dynamics, and elicited immune response, which may instruct future research on antiviral strategies.

Natural reversion promotes LPS elongation in an attenuated Coxiella burnetii strain

Lipopolysaccharide (LPS) phase variation is a critical aspect of virulence in many Gram-negative bacteria. It is of particular importance to Coxiella burnetii, the biothreat pathogen that causes Q fever, as in vitro propagation of this organism leads to LPS truncation, which is associated with an attenuated and exempted from select agent status (Nine Mile II, NMII). Here, we demonstrate that NMII was recovered from the spleens of infected guinea pigs. Moreover, these strains exhibit a previously unrecognized form of elongated LPS and display increased virulence in comparison with the initial NMII strain. The reversion of a 3-bp mutation in the gene cbu0533 directly leads to LPS elongation. To address potential safety concerns, we introduce a modified NMII strain unable to produce elongated LPS.

Production and characterization of anti-porcine CXCL10 monoclonal antibodies

Research on C-X-C motif chemokine ligand 10 (CXCL10) has been widely reported for humans and select animal species, yet immune reagents are limited for pig chemokines. Our goal is to provide veterinary immunologists and the biomedical community with new commercial immune reagents and standardized assays. Recombinant porcine CXCL10 (rPoCXCL10) protein was produced by yeast expression and used to generate a panel of α CXCL10 monoclonal antibodies (mAbs). All mAbs were assessed for cross-inhibition and reactivity to orthologous yeast expressed CXCL10 proteins. Characterization of a panel of nine α PoCXCL10 mAbs identified six distinct antigenic determinants. A sensitive quantitative sandwich ELISA was developed with anti-PoCXCL10-1.6 and −1.9 mAb; reactivity was verified with both rPoCXCL10 and native PoCXCL10, detected in supernatants of peripheral blood mononuclear cells stimulated with rPoIFNγ or PMA/Ionomycin. Immunostaining of in vitro rPoIFNγ stimulated pig spleen and blood cells verified CXCL10 + cells as CD3-CD4-CD172+, with occasional CD3-CD4 + CD172 + subsets. Comparison studies determined that α PoCXCL10-1.4 mAb was the ideal mAb clone for intracellular staining, whereas with α PoCXCL10-1.1 and −1.2 mAbs were best for immunohistochemistry analyses. These techniques and tools will be useful for evaluating swine immune development, responses to infectious diseases and vaccines, as well as for improving utility of pigs as an important biomedical model.

Comprehensive assessment of peptide derived from pig spleen: Preparation, bioactivity and structure-activity relationships

Pig spleens, as the by-products of pig resources, were often discarded as waste or used for low-value. Therefore, we tried to isolate bioactive peptide from protein hydrolysate of pig spleens to increase their added value. In this study, the low molecular weight peptides (<3000 Da) were prepared by enzyme hydrolysis and its structure-activity relationship was discussed. The main secondary structures of the pig spleen peptide (PSP) were β-turn and random coil by Fourier transform infrared (FT-IR) analysis. Hydrophobic amino acids accounted for half of the total amino acid content. In vitro, PSP had no inhibition effect on HepG2, HCT116, H22 and S180 cells. While PSP exhibited scavenging capacities of PSP on 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion (O2−) and hydroxyl (OH) radicals. In vivo, the activity of antioxidant enzymes (superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX)) increased and the level of malondialdehyde (MDA) decreased in PSP-treated mice. PSP displayed obvious immunomodulatory effects including stimulation of multiple immune cells to proliferate, production of cytokines (IFN-γ, IL-2, IL-4 and TNF-α) and increase the level of IgG, IgM and IgA antibodies. It induced solid tumor cells apoptosis and arrested at S phase of cell cycle. In conclusion, PSP had significant immunomodulatory effects and antioxidant activity. These results provided theoretical and data support for bioactive peptides derived from meat by-products used as the functional foods.

Preliminary investigation of organ-protective effects of fullerenol in vivo in acute toxicity during doxorubicin therapy in healthy pigs

Abstract Objectives The potential protective effect of fullerenol [C60(OH)24, Frl], a strong antioxidant, was investigated at a single dose of 25 mg/kg, administered orally (per os; p.o.) and intraperitoneally (i.p.), in the liver, lung, kidney, heart and spleen of healthy pigs, prior to doxorubicin (Dox)-induced toxicity at a single i.p. dose of 10 mg/kg. Methods We used the F1 generation of an in-vivo pig model (whose parents were Swedish Landrace and Large Yorkshire) to explore whether Frl, administered 6 h for p.o. and 30 min for i.p. prior to Dox treatment, could protect organs against damage caused by oxidative stress. Key findings According to the macroscopic, haematological, biochemical and physiological results, Frl exerted a potential protective effect on all investigated organs (heart, liver, lung, spleen and kidney) in pigs after i.p. administration. However, the results from Frl p.o. administration were inconclusive, therefore warranting further investigation. Conclusions Frl at a low dose of 25 mg/kg demonstrated satisfactory organ protection against Dox-induced toxicity in healthy pigs after i.p. injection, However, additional follow-up studies in pig models of cancer and with longer Dox and Frl treatment periods would be required to further delineate the optimal and clinically relevant conditions for Frl administration.

Comprehensive Analysis of CircRNA Expression Profiles in Multiple Tissues of Pigs.

Circular RNAs (circRNAs) are a class of non-coding RNAs with diverse functions, and previous studies have reported that circRNAs are involved in the growth and development of pigs. However, studies about porcine circRNAs over the past few years have focused on a limited number of tissues. Based on 215 publicly available RNA sequencing (RNA-seq) samples, we conducted a comprehensive analysis of circRNAs in nine pig tissues, namely, the gallbladder, heart, liver, longissimus dorsi, lung, ovary, pituitary, skeletal muscle, and spleen. Here, we identified a total of 82,528 circRNAs and discovered 3818 novel circRNAs that were not reported in the CircAtlas database. Moreover, we obtained 492 housekeeping circRNAs and 3489 tissue-specific circRNAs. The housekeeping circRNAs were enriched in signaling pathways regulating basic biological tissue activities, such as chromatin remodeling, nuclear-transcribed mRNA catabolic process, and protein methylation. The tissue-specific circRNAs were enriched in signaling pathways related to tissue-specific functions, such as muscle system process in skeletal muscle, cilium organization in pituitary, and cortical cytoskeleton in ovary. Through weighted gene co-expression network analysis, we identified 14 modules comprising 1377 hub circRNAs. Additionally, we explored circRNA-miRNA-mRNA networks to elucidate the interaction relationships between tissue-specific circRNAs and tissue-specific genes. Furthermore, our conservation analysis revealed that 19.29% of circRNAs in pigs shared homologous positions with their counterparts in humans. In summary, this extensive profiling of housekeeping, tissue-specific, and co-expressed circRNAs provides valuable insights into understanding the molecular mechanisms of pig transcriptional expression, ultimately deepening our understanding of genetic and biological processes.

Hydroxy Selenomethionine Exert Different Protective Effects Against Dietary Oxidative Stress-Induced Inflammatory Responses in Spleen and Thymus of Pigs.

Oxidative stress (OS) is widespread in animal husbandry, which causes edema in immune organs and suppresses immune function of animals. Selenium (Se) is an essential trace element involved in immune regulation and improves animals' immunity. In present study,growing and finishing pigs were used to determine the protective effects of the new organic Se (hydroxy selenomethionine, OH-SeMet) on dietary oxidative stress (DOS) induced inflammatory responses, and the corresponding response of selenotranscriptome in spleen and thymus. Forty castrated male pigs (25.0 ± 3.0 kg) were randomly grouped into 5 dietary treatments (n = 8) and fed on basal diet (formulated with normal corn and normal oils) or oxidized diet (formulated with aged corn and oxidized oils) supplied with 0.0, 0.3, 0.6, or 0.9 mg Se/kg OH-SeMet, after 16 weeks, the corresponding indicators were determined. Results showed thatDOS moderately increased the spleen and thymus index, decreased the antioxidant capacity of serum, spleen and thymus, and increased the concentration of serum inflammatory cytokines (IL-6 and TNF-α). The inflammatory response in spleen and thymus under DOS were discrepancies, DOS increased the expression of inflammation-related gene (IFN-β and TNF-α) in thymus, while exhibited no impact on that of thespleen. Dietary OH-SeMet supplementation exhibited protective effects, which decreased the spleen and thymus index, improved the antioxidant capacity of serum, spleen and thymus, and decreased the serum IL-1β and IL-6 levels. Se supplementation exhibited limited impact on the inflammation-related genes in spleen, except decreased the mRNA expression of IL-8. On the contrary, Se supplementation showed more impact on that of the thymus, which decreased the mRNA expression of IL-8 and TNF-α, increased the expression of IFN-β, IL-6, IL-10, and MCP1. In addition, selenotranscriptome responsive to dietary Se levels in spleen and thymus were discrepancies. Se supplementation increased the mRNA expression of theselenotranscriptome in thymus, while exhibited limited impact on that of in spleen. In conclusion,dietary OH-SeMet supplementation mitigates the DOS-induced immunological stress by increasing the antioxidant capacity and altering the expression of inflammation-related genes and selenotranscriptome in immune organs, and these response in spleen and thymus were discrepancies.

Molecular and biological properties of the African swine fever virus (Asfarviridae: Asfivirus) isolate ASF/Tatarstan 20/WB-12276

Up-to-date data and full characterization of circulating ASFV isolates play a crucial role in virus eradication and control in endemic regions and countries. The aim of the study was to evaluate and characterize the molecular and biological properties of the ASFV isolate ASF/Tatarstan 20/WB-12276, conduct phylogenetic analysis, and compare the results with isolates circulating in Europe and Asia. For bioassay, eight heads of the Large White pigs weighing 15-20 kg/head were used. Detection of specific anti-ASFV antibodies by ELISA and immunoperoxidase method. Detection of ASFV genome was performed by qPCR. Isolation of ASF/Tatarstan 20/WB-12276 and determination of titer were performed in pig spleen cell culture. Sequencing was carried out by the Sanger method. The virus was characterized as highly virulent and capable of causing acute to subacute forms of ASF. Phylogenetic analysis revealed substitutions in the genome of the ASF/Tatarstan 20/WB-12276 isolate (IGR/I73R-I329L and I267L markers) that supported the clustering of the studied variant with isolates prevalent in most of Europe and Asia. For the first time, the molecular and biological properties of the ASF/Tatarstan 20/WB-12276 virus isolate taken from a wild boar shot on the territory of the Republic of Tatarstan were studied and analyzed.

Genome-wide analysis of circRNA regulation during spleen development of Chinese indigenous breed Meishan pigs

BackgroundNumerous circular RNAs (circRNAs) have been recently identified in porcine tissues and cell types. Nevertheless, their significance in porcine spleen development is yet unelucidated. Herein, we reported an extensive overlook of circRNA expression profile during spleen development in Meishan pigs.ResultsOverall, 39,641 circRNAs were identified from 6,914 host genes. Among them, many circRNAs are up- or down-regulated at different time points of pig spleen development. Using WGCNA analysis, we revealed two essential modules for protein-coding genes and circRNAs. Subsequent correlation analysis revealed 67 circRNAs/co-expressed genes that participated in immnue-associated networks. Furthermore, a competing endogenous RNA (ceRNA) network analysis of circRNAs revealed that 12 circRNAs modulated CD226, MBD2, SAMD3, SIT1, SRP14, SYTL3 gene expressions via acting as miRNA sponges. Moreover, the circRNA_21767/miR-202-3p axis regulated SIT1 expression in a ceRNA manner, which is critical for the immune-based regulation of spleen development in Meishan pigs.ConclusionsOverall, our results demonstrated that the circRNAs were differentially expressed during different stages of porcine spleen development, meanwhile the circRNAs interacted with immune-related genes in a ceRNA-based fashion. Moreover, we presented biomedical researchers with RNAseqTools, a user-friendly and powerful software for the visualization of transcriptome profile data.

Metabolomic analysis of pig spleen reveals African swine fever virus infection increased acylcarnitine levels to facilitate viral replication.

African swine fever (ASF) is a devastating disease caused by the African swine fever virus (ASFV) that adversely affects the pig industry. The spleen is the main target organ of ASFV; however, the function of metabolites in the spleen during ASFV infection is yet to be investigated. To define the metabolic changes in the spleen after ASFV infection, untargeted and targeted metabolomics analyses of spleens from ASFV-infected pigs were conducted. Untargeted metabolomics analysis revealed 540 metabolites with significant differential levels. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that these metabolites were mainly enriched in metabolic pathways, including nucleotide metabolism, purine metabolism, arginine biosynthesis, and neuroactive ligand-receptor interaction. Moreover, 134 of 540 metabolites quantified by targeted metabolomics analysis had differential levels and were enriched in metabolic pathways such as the biosynthesis of cofactors, ABC transporters, and biosynthesis of amino acids. Furthermore, coalition analysis of untargeted and targeted metabolomics data revealed that the levels of acylcarnitines, which are intermediates of fatty acid β-oxidation, were significantly increased in ASFV-infected spleens compared with those in the uninfected spleens. Moreover, inhibiting fatty acid β-oxidation significantly reduced ASFV replication, indicating that fatty acid β-oxidation is essential for this process. To our knowledge, this is the first report presenting the metabolite profiles of ASFV-infected pigs. This study revealed a new mechanism of ASFV-mediated regulation of host metabolism. These findings provide new insights into the pathogenic mechanisms of ASFV, which will benefit the development of target drugs for ASFV replication. IMPORTANCE African swine fever virus, the only member of the Asfarviridae family, relies on hijacking host metabolism to meet the demand for self-replication. However, the change in host metabolism after African swine fever virus (ASFV) infection remains unknown. Here, we analyzed the metabolic changes in the pig spleen after ASFV infection for the first time. ASFV infection increased the levels of acylcarnitines. Inhibition of the production and metabolism of acylcarnitines inhibited ASFV replication. Acylcarnitines are the vital intermediates of fatty acid β-oxidation. This study highlights the critical role of fatty acid β-oxidation in ASFV infection, which may help identify target drugs to control African swine fever disease.

Mitochondria-mediated apoptosis and endoplasmic reticulum stress are involved in the toxicity induced by copper in the porcine spleen.

Copper (Cu) is one of the common heavy metal pollutants in the environment, and its toxic mechanisms have been extensively studied. However, the immunotoxicity induced by Cu remains rarely reported, and the effects of Cu on endoplasmic reticulum stress and mitochondria-mediated apoptosis have been little studied in the spleen. In this study, pigs were fed with different contents of Cu (10, 125, and 250mg/kg Cu) for 80days to establish a toxicity model. The results showed the Cu exposure triggered endoplasmic reticulum stress in the spleen, as evidenced by increased mRNA and protein levels of GRP94, GRP78, CHOP, XBP1, ATF6, and JNK; the positive rate of GRP78 increased by immunofluorescence analysis. Additionally, mitochondrial fission and fusion homeostasis were disrupted, the expression levels of mitochondrial dynamics-related genes Mfn1, Mfn2, and OPA1 decreased, DRP1 increased, and the positive rate of Mfn1 decreased by immunofluorescence analysis. Furthermore, Cu exposure could induce apoptosis, as demonstrated by the increased expression level of related proteins and genes Bak, Bax, Caspase-3, P53, and Cytc. In conclusion, these results suggest chronic Cu exposure can lead to endoplasmic reticulum stress and imbalance in mitochondrial dynamics and induced apoptosis of pig spleen, and these results provided new insights into the underlying mechanism of Cu exposure caused splenic toxicity, which has public health implications where humans and animals are exposed to copper contamination.

Features of accumulation and variability of stable strontium in some organs and tissues of landrace pigs

The authors in the article demonstrated the results of assessing the level of stable strontium in the skeletal muscles, liver, kidneys and spleen of Landrace pigs at the end of the technological cycle. The studies were performed on clinically healthy animals raised in a large pig farm in the Altai Territory. The conditions of keeping the animals corresponded to those typical for meat fattening up to 100 kg. Feeding was carried out with complete feed. Elemental analysis of samples of parenchymal organs and muscle tissue was performed using inductively coupled plasma atomic emission spectral analysis. Data processing was performed using Microsoft Office Excel and the R programming language in the data analysis environment RStudio version 2022.07.2+576 (RStudio, PBC). The distribution did not correspond to normal in all cases. The dispersions are not homogeneous. Based on the mean value and median, an increasing ranked series of strontium content in organs and tissues were established: liver = skeletal muscles &lt; spleen &lt; kidneys, in numerical terms: 1: 1: 3.7: 4.7 and 1: 1: 3.9: 5.7 respectively. Medians for stable strontium in the liver, kidney, spleen, and skeletal muscle were 0.024; 0.130; 0.089 and 0.023 mg/kg, respectively. Reference intervals for the liver are 0.004-0.043 and for the spleen - 0.030-0.145 mg/kg. The most significant range of variability is characteristic of the kidneys of pigs. A considerable uniformity is typical of the liver and skeletal muscles. Using the Kruskal-Wallis test, it was established that the accumulation of stable strontium significantly differs in the examined organs and tissues of pigs (H = 68.9, df = 3, p &lt; 0.0001). Pairwise comparison showed significant differences for the kidney-liver, spleen-liver, skeletal muscle-kidney, and skeletal muscle-spleen pairs. Two clusters were identified according to the similarity of strontium accumulation: liver and skeletal muscles, kidneys and spleen. The calcium-strontium ratio, taking into account medians, for skeletal muscles was 1:1833, and for the liver - 1:1870. The established results can serve as an approximate physiological norm for the concentration of stable strontium in individual structures of the organism of Landrace pigs under the conditions of Western Siberia.

Cadmium exposure induces necroptosis of porcine spleen via ROS-mediated activation of STAT1/RIPK3 signaling pathway.

Cadmium (Cd), a heavy metal, is used in a wide range of applications, such as plastics, electroplating process, electronics, and so forth. Due to its bioaccumulation ability, Cd can contaminate soil, water, air and food. To determine the effect of Cd exposure on the necroptosis in pig spleen and its mechanistic investigation, we constructed a model in pigs by feeding them food containing 20 mg/kg Cd. In this study, we analyzed the effects of Cd exposure on pig spleen through HE staining, Quantitative real-time PCR (qRT-PCR), Western blot (WB), and principal component analysis (PCA). Results show that Cd exposure can destroy the structure and function of pig spleen, which is closely related to necroptosis. Further results show that Cd exposure can induce necroptosis through ROS-mediated activation of Signal transducer and activator of transcription 1/Receptor-Interacting Serine/Threonine-Protein Kinase 3 (STAT1/RIPK3) signaling pathway in pig spleen. Additionally, Cd exposure also can affect the stability of mitochondrial-associated endoplasmic reticulum membrane (MAMs) structure, which also contributes to the process of necroptosis. Our study provides insights into the physiological toxicity caused by Cd exposure.

Development of Chimeric Hepatitis B (HBV) – Norovirus (NoV) P particle as candidate vaccine against Hepatitis B and norovirus infection

IntroductionHepatitis B remains a global problem with no effective treatment. Here, a mucosal vaccine candidate was developed with HBsAg and HBcAg, to provide both prophylactic and therapeutic protection against hepatitis B. The antigens were presented using the P particle of human norovirus (HuNov). As a result, the chimeric HBV – HuNoV P particle can act as a dual vaccine for hepatitis B and HuNoV. MethodsThe vaccine candidate was expressed and purified from Escherichia coli BL21 (DE3) cells. HBV-HuNoV chimeric P particles were successfully expressed and isolated, with sizes of approximately 25.64 nm. Then, the HBV-HuNoV chimeric P particles were evaluated for safety and immunogenicity in mice and gnotobiotic (Gn) pigs. After three doses (5 µg/dose in mice and 200 µg/dose in Gn pigs) of intranasal immunization, humoral and cellular immune responses, as well as toxicity, were evaluated. ResultsThe vaccine candidate induced strong HBV-HuNoV specific IFN-γ producing T-cell responses in the ileum, spleen, and blood of Gn pigs. Serum IgG and IgA antibodies against HBV-HuNoV chimeric P particles also increased significantly in Gn pigs. Increased HBsAg- and HuNoV-specific serum IgG responses were observed in mice and Gn pigs, although not statistically significant. The vaccine candidate did not show any toxicity in mice. ConclusionsIn summary, the chimeric HBV-HuNoV P particle vaccine given intranasally was safe and induced strong cellular and humoral immune responses in Gn pig. Modifications to the vaccine structure and dosage need to be evaluated in future studies to further enhance immunogenicity and induce more balanced humoral and cellular responses.