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1.
Virology ; 539: 38-48, 2020 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-31670218

RESUMO

Ionic calcium (Ca2+) is a versatile intracellular second messenger that plays important roles in cellular physiological and pathological processes. Porcine deltacoronavirus (PDCoV) is an emerging enteropathogenic coronavirus that causes serious vomiting and diarrhea in suckling piglets. In this study, the role of Ca2+ to PDCoV infection was investigated. PDCoV infection was found to upregulate intracellular Ca2+ concentrations of IPI-2I cells. Chelating extracellular Ca2+ by EGTA inhibited PDCoV replication, and this inhibitory effect was overcome by replenishment with CaCl2. Treatment with Ca2+ channel blockers, particularly the L-type Ca2+ channel blocker diltiazem hydrochloride, inhibited PDCoV infection significantly. Mechanistically, diltiazem hydrochloride reduces PDCoV infection by inhibiting the replication step of the viral replication cycle. Additionally, knockdown of CACNA1S, the L-type Ca2+ voltage-gated channel subunit, inhibited PDCoV replication. The combined results demonstrate that PDCoV modulates calcium influx to favor its replication.


Assuntos
Cálcio/metabolismo , Infecções por Coronavirus/metabolismo , Infecções por Coronavirus/virologia , Coronavirus/fisiologia , Doenças dos Suínos/metabolismo , Doenças dos Suínos/virologia , Replicação Viral , Animais , Sinalização do Cálcio , Suínos , Porco Miniatura
2.
Vet Res ; 50(1): 111, 2019 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-31864417

RESUMO

Interleukin-11 (IL-11), a well-known anti-inflammatory factor, provides protection from intestinal epithelium damage caused by physical or chemical factors. However, little is known of the role of IL-11 during viral infections. In this study, IL-11 expression at mRNA and protein levels were found to be high in Vero cells and the jejunum of piglets during porcine epidemic diarrhea virus (PEDV) infection, while IL-11 expression was found to be positively correlated with the level of viral infection. Pretreatment with recombinant porcine IL-11 (pIL-11) was found to suppress PEDV replication in Vero E6 cells, while IL-11 knockdown promoted viral infection. Furthermore, pIL-11 was found to inhibit viral infection by preventing PEDV-mediated apoptosis of cells by activating the IL-11/STAT3 signaling pathway. Conversely, application of a STAT3 phosphorylation inhibitor significantly antagonized the anti-apoptosis function of pIL-11 and counteracted its inhibition of PEDV. Our data suggest that IL-11 is a newfound PEDV-inducible cytokine, and its production enhances the anti-apoptosis ability of epithelial cells against PEDV infection. The potential of IL-11 to be used as a novel therapeutic against devastating viral diarrhea in piglets deserves more attention and study.


Assuntos
Infecções por Coronavirus/veterinária , Interleucina-11/metabolismo , Vírus da Diarreia Epidêmica Suína , Doenças dos Suínos/virologia , Animais , Antivirais , Apoptose , Clonagem Molecular , Infecções por Coronavirus/metabolismo , Infecções por Coronavirus/virologia , Regulação da Expressão Gênica/imunologia , Interleucina-11/genética , Receptores de Interleucina-11/genética , Receptores de Interleucina-11/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/metabolismo , Células Vero
3.
Emerg Microbes Infect ; 8(1): 1642-1657, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31711377

RESUMO

Diverse noroviruses infect humans and animals via the recognition of host-specific glycan ligands. Genogroup II (GII) noroviruses consist of human noroviruses (huNoVs) that generally bind histo-blood group antigens (HBGAs) as host factors and three porcine norovirus (porNoV) genotypes (GII.11/18/19) that form a genetic lineage lacking HBGA-binding ability. Thus, these GII porNoVs provide an excellent model to study norovirus evolution with host ligand specificity changes. Here we solved the crystal structures of a native GII.11 porNoV P protein and a closely-related GII.3 huNoV P protein complexed with an HBGA, focusing on the HBGA-binding sites (HBSs) compared with the previously known ones to understand the structural basis of the host ligand specificity change. We found that the GII.3 huNoV binds HBGAs via a conventional GII HBS that uses an arginine instead of the conserved aromatic residue for the required Van der Waals interaction, while the GII.11 porNoV HBS loses its HBGA-binding function because of two mutations (Q355/V451). A mutant that reversed the two mutated residues back to the conventional A355/Y451 restored the HBGA-binding function of the GII.11 porNoV P protein, which validated our observations. Similar mutations are also found in GII.19 porNoVs and a GII.19 P protein mutant with double reverse mutations restored the HBS function. This is the first reconstruction of a functional HBS based on one with new host specificity back to its parental one. These data shed light on the molecular basis of structural adaptation of the GII porNoVs to the pig hosts through mutations at their HBSs.


Assuntos
Antígenos de Grupos Sanguíneos/metabolismo , Infecções por Caliciviridae/veterinária , Infecções por Caliciviridae/virologia , Norovirus/metabolismo , Doenças dos Suínos/virologia , Proteínas Virais/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Antígenos de Grupos Sanguíneos/genética , Infecções por Caliciviridae/genética , Infecções por Caliciviridae/metabolismo , Genótipo , Humanos , Ligantes , Mutação , Norovirus/química , Norovirus/classificação , Norovirus/genética , Filogenia , Ligação Proteica , Alinhamento de Sequência , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/metabolismo , Proteínas Virais/química , Proteínas Virais/genética
4.
Food Funct ; 10(8): 5152-5165, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31373594

RESUMO

The aim of this study was to investigate the effects of dietary ß-hydroxy-ß-methylbutyrate (HMB) on lipopolysaccharide (LPS)-induced muscle atrophy and to investigate the mechanisms involved. Sixty pigs (21 ± 2 days old, 5.86 ± 0.18 kg body weight) were used in a 2 × 3 factorial design and the main factors included diet (0, 0.60%, or 1.20% HMB) and immunological challenge (LPS or saline). After 15 d of treatment with LPS and/or HMB, growth performance, blood parameters, and muscle protein degradation rate were measured. The results showed that in LPS-injected pigs, 0.60% HMB supplementation increased the average daily gain and average daily feed intake and decreased the feed : gain ratio (P < 0.05), with a concurrent increase of lean percentage. Moreover, 0.60% HMB supplementation decreased the serum concentrations of blood urea nitrogen, IL-1ß, and TNF-α and the rate of protein degradation as well as cell apoptosis in selected muscles (P < 0.05). In addition, dietary HMB supplementation (0.60%) regulated the expression of genes involved in mitochondrial biogenesis and increased the phosphorylation of Akt and Forkhead Box O3a (FoxO3a) in selected muscles, accompanied by decreased protein expression of muscle RING finger 1 and muscle atrophy F-box. These results indicate that HMB may exert protective effects against LPS-induced muscle atrophy by normalizing the Akt/FoxO3a axis that regulates ubiquitin proteolysis and by improving mitochondrial biogenesis.


Assuntos
Proteína Forkhead Box O3/metabolismo , Mitocôndrias/efeitos dos fármacos , Proteínas Musculares/metabolismo , Atrofia Muscular/veterinária , Proteínas Proto-Oncogênicas c-akt/metabolismo , Doenças dos Suínos/prevenção & controle , Valeratos/administração & dosagem , Ração Animal/análise , Animais , Feminino , Proteína Forkhead Box O3/genética , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolissacarídeos/efeitos adversos , Masculino , Mitocôndrias/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Atrofia Muscular/induzido quimicamente , Atrofia Muscular/metabolismo , Atrofia Muscular/prevenção & controle , Biogênese de Organelas , Proteólise/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/efeitos dos fármacos , Suínos , Doenças dos Suínos/induzido quimicamente , Doenças dos Suínos/genética , Doenças dos Suínos/metabolismo
5.
J Therm Biol ; 84: 375-383, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31466777

RESUMO

Bile acids (BAs) are critical for cholesterol homeostasis and new roles in metabolism and endocrinology have been demonstrated recently. It remains unknown whether BA metabolism can be affected by heat stress (HS). The objective of this study was to describe the shifts in serum, hepatic and intestinal BA profiles induced by chronic HS. Twenty-seven Large White pigs weighing 40.8 ±â€¯2.7 kg were assigned to one of the three treatments: a control group (CON, 23 °C), a HS group (33 °C), or a pair-fed group (PF, 23 °C and fed the same amount as HS group) for 21 d. The concentrations of taurine-conjugated BAs (TUDCA and THDCA in serum and TCDCA, TUDCA, THDCA and THCA in liver) were decreased in HS and PF pigs. However, in HS pigs, a reduction in taurine-conjugated BAs (TCBA) correlated with decreased liver genes expression of BA synthesis, conjugation and uptake transport. BA regulated-genes (FXR, TGR5 and FGFR4) in HS pigs and TGR5, FGFR4 and KLß in PF pigs were down-regulated in liver. In ileum, total BAs and glycoursodeoxycholic acid concentrations were higher in HS pigs than other groups and PF group, respectively (P < 0.05). TCBA (P = 0.01) and tauroursodeoxycholic acid (P < 0.01) were decreased in PF group. BA transporters (OSTα and MRP3) were up-regulated in HS pigs compared with CON and PF pigs, respectively (P < 0.01). In cecum, ursodeoxycholic acid was higher in HS (P = 0.02) group than CON group. The expression of apical sodium-coupled bile acid transporter (P = 0.04) was lower in HS pigs than CON pigs, while OSTß (P < 0.01) was greater in HS group than PF group. These results suggest that chronic HS suppressed liver activity of synthesis and uptake of TCBA, at least in part, which was independent of reduced feed intake.


Assuntos
Ácidos e Sais Biliares/metabolismo , Transtornos de Estresse por Calor/metabolismo , Doenças dos Suínos/metabolismo , Suínos/metabolismo , Animais , Ácidos e Sais Biliares/sangue , Conteúdo Gastrointestinal/química , Conteúdo Gastrointestinal/microbiologia , Transtornos de Estresse por Calor/veterinária , Resposta ao Choque Térmico , Temperatura Alta , Fígado/metabolismo , Masculino
6.
Res Vet Sci ; 125: 227-234, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31284225

RESUMO

The aim of this study was to identify biological pathways and proteins differentially expressed in saliva of pigs in two conditions of compromised welfare: an acute stress consisting of restraint with a nose snare and in pigs with lameness which is a highly frequent problem in the swine industry. For this purpose, high-resolution quantitative proteomics based on Tandem Mass Tags labelling was used. Four proteins showed significant differences in the conditions of compromised welfare, namely cornulin, the heat shock protein 27 and lactate dehydrogenase (LDH), that showed significant increases, whereas immunoglobulin J chain showed a significant decrease. LDH, which was the protein that showed the highest differences, was selected for validation and clinical evaluation as a diagnostic biomarker. Significant changes in this protein were observed between pigs restrained with a nose snare and pigs with lameness compared with healthy pigs when measured with available commercial assays in a larger population of pigs. In conclusion, this study reports that in situations of compromised welfare on farm, such as acute stress and lameness in pigs, there are changes in proteins and metabolic pathways in saliva, and describes a series of proteins that could potentially be used as biomarkers for both short term acute stress and longer term chronic stress of lameness. These biomarkers would have the advantage of being measured in saliva by a noninvasive and not stressful collection sampling procedure.


Assuntos
Bem-Estar do Animal , Coxeadura Animal/metabolismo , Saliva/química , Proteínas e Peptídeos Salivares/química , Doenças dos Suínos/diagnóstico , Animais , Biomarcadores/química , Biomarcadores/metabolismo , Proteômica , Proteínas e Peptídeos Salivares/metabolismo , Suínos , Doenças dos Suínos/metabolismo
7.
Int J Mol Sci ; 20(14)2019 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-31330869

RESUMO

(1) Background: Vitamin D (VD) plays a vital role in anti-viral innate immunity. However, the role of VD in anti-rotavirus and its mechanism is still unclear. The present study was performed to investigate whether VD alleviates rotavirus (RV) infection through a microRNA-155-5p (miR-155-5p)-mediated regulation of TANK-binding kinase 1 (TBK1)/interferon regulatory factors 3 (IRF3) signaling pathway in vivo and in vitro. (2) Methods: The efficacy of VD treatment was evaluated in DLY pig and IPEC-J2. Dual-luciferase reporter activity assay was performed to verify the role of miR-155-5p in 1α,25-dihydroxy-VD3 (1,25D3) mediating the regulation of the TBK1/IRF3 signaling pathway. (3) Results: A 5000 IU·kg-1 dietary VD3 supplementation attenuated RV-induced the decrease of the villus height and crypt depth (p < 0.05), and up-regulated TBK1, IRF3, and IFN-ß mRNA expressions in the jejunum (p < 0.05). Incubation with 1,25D3 significantly decreased the RV mRNA expression and the RV antigen concentration, and increased the TBK1 mRNA and protein levels, and the phosphoprotein IRF3 (p-IRF3) level (p < 0.05). The expression of miR-155-5p was up-regulated in response to an RV infection in vivo and in vitro (p < 0.05). 1,25D3 significantly repressed the up-regulation of miR-155-5p in vivo and in vitro (p < 0.05). Overexpression of miR-155-5p remarkably suppressed the mRNA and protein levels of TBK1 and p-IRF3 (p < 0.01), while the inhibition of miR-155-5p had an opposite effect. Luciferase activity assays confirmed that miR-155-5p regulated RV replication by directly targeting TBK1, and miR-155-5p suppressed the TBK1 protein level (p < 0.01). (4) Conclusions: These results indicate that miR-155-5p is involved in 1,25D3 mediating the regulation of the TBK1/IRF3 signaling pathway by directly targeting TBK1.


Assuntos
Fator Regulador 3 de Interferon/metabolismo , MicroRNAs/genética , Proteínas Serina-Treonina Quinases/metabolismo , Infecções por Rotavirus/veterinária , Rotavirus/fisiologia , Transdução de Sinais/efeitos dos fármacos , Vitamina D/farmacologia , Animais , Regulação da Expressão Gênica , Rotavirus/efeitos dos fármacos , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/metabolismo , Doenças dos Suínos/virologia , Replicação Viral/efeitos dos fármacos
8.
Vet Res ; 50(1): 48, 2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31221216

RESUMO

Enterotoxigenic Escherichia coli (ETEC) are an important cause of post-weaning diarrhea (PWD) in piglets. The IL-17 cytokine family is well known to play important roles in the host defense against bacterial infections at the mucosa. Previously, we reported the potential role of IL-17A in clearing an ETEC infection in piglets. IL-17C, another member of the IL-17 family, is highly expressed in the intestinal epithelium, however, its role during an ETEC infection is still unclear. In this study, we demonstrate that F4+ ETEC induce IL-17C mRNA and protein expression in intestinal tissues as well as in porcine intestinal epithelial cells (IPEC-J2). This IL-17C production is largely dependent on TLR5 signaling in IPEC-J2 cells. Both F4+ ETEC infection and exogenous IL-17C increased the expression of antimicrobial peptides and tight junction proteins, such as porcine beta-defensin (pBD)-2, claudin-1, claudin-2 and occludin in IPEC-J2 cells. Taken together, our data demonstrate that TLR5-mediated IL-17C expression in intestinal epithelial cells enhances mucosal host defense responses in a unique autocrine/paracrine manner in the intestinal epithelium against ETEC infection.


Assuntos
Escherichia coli Enterotoxigênica/fisiologia , Infecções por Escherichia coli/veterinária , Interleucina-17/genética , Doenças dos Suínos/genética , Receptor 5 Toll-Like/genética , Animais , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/metabolismo , Infecções por Escherichia coli/microbiologia , Interleucina-17/metabolismo , Mucosa Intestinal/fisiopatologia , Suínos , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia , Receptor 5 Toll-Like/metabolismo
9.
Food Funct ; 10(6): 3535-3542, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31149689

RESUMO

Enteric infection is a major cause of morbidity and mortality in both humans and animals worldwide. Immunotherapy against intestinal infection is a well-known alternative to the antibiotic strategy. Herein, we demonstrated that isoleucine significantly suppressed the multiplication of E. coli in the presence of IPEC-J2 cells. Isoleucine supplementation enhanced the concentrations of total plasma protein and IgA in pigs compared to the alanine control diet, while inhibiting the increase in plasma endotoxin and IL-6 contents induced by E. coli challenge. A significant interaction between the E. coli challenge and the diet treatment was found in the red blood cell volume. Isoleucine improved the expression of porcine ß-defensin-1 (pBD-1), pBD-2, pBD-3, pBD-114 and pBD-129 in the jejunum and ileum of pigs with or without E. coli challenge. Conclusively, isoleucine attenuated the infection caused by the E. coli challenge possibly through increasing the intestinal ß-defensin expression and inhibiting the increase in plasma endotoxin and IL-6 in weaned pigs.


Assuntos
Defensinas/genética , Endotoxinas/sangue , Infecções por Escherichia coli/veterinária , Escherichia coli/fisiologia , Interleucina-6/sangue , Mucosa Intestinal/metabolismo , Isoleucina/administração & dosagem , Doenças dos Suínos/tratamento farmacológico , Animais , Defensinas/metabolismo , Suplementos Nutricionais/análise , Escherichia coli/efeitos dos fármacos , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/metabolismo , Infecções por Escherichia coli/microbiologia , Íleo/efeitos dos fármacos , Íleo/metabolismo , Íleo/microbiologia , Interleucina-6/genética , Mucosa Intestinal/microbiologia , Jejuno/efeitos dos fármacos , Jejuno/metabolismo , Jejuno/microbiologia , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia
10.
Food Funct ; 10(5): 2926-2934, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31070611

RESUMO

Weaning stress in piglets can lead to poor health outcomes and reduced production. We investigated the effects of probiotics, one potential antibiotic alternative, on the growth performance, serum biochemical parameters, intestinal morphology, mucosal immunity, hypothalamic neurotransmitters, and colonic microflora in weaned piglets. Thirty-six weaned piglets were fed a basal diet, a diet supplemented with colistin sulphate antibiotic, or a diet supplemented with probiotics including Clostridium butyricum, Bacillus subtilis, and B. licheniformis. Probiotics significantly increased the feed : gain ratio, improved the average day gain from day 1 to day 28, and decreased the diarrhoea index. Probiotics also lowered the serum concentrations of AST, ALT, and ALP on day 14 and lowered the serum concentration of ALT on day 28 compared with the control. Probiotic supplementation caused fewer ileal apoptotic cells. The serum and ileal concentrations of TNF-α and IL-1ß on day 28 were significantly lowered, and the serum concentrations of IL-6 were significantly lowered on days 14 and 28. Probiotic-fed piglets exhibited higher contents of hypothalamic serotonin and dopamine as well as serum γ-aminobutyric acid along with higher colonic concentrations of butyrate and valerate on day 28. High-throughput sequencing showed 972 core operational taxonomic units among all groups, of which 48 were unique to the probiotic-treated group. The relative abundance of genus Bacillus and species Bacillus velezensis was enriched in probiotic piglets; the phylogenetic investigation of communities by the reconstruction of unobserved states indicated that amino acid metabolism, DNA repair, replication and recombination proteins, and secretion systems were enriched with probiotics. In conclusion, the Clostridium butyricum-based probiotics improved growth performance, enhanced intestinal morphology, changed hypothalamic neurotransmitters and modulated colonic microflora in weaned piglets.


Assuntos
Clostridium butyricum/fisiologia , Colo/microbiologia , Diarreia/veterinária , Hipotálamo/metabolismo , Intestinos/imunologia , Neurotransmissores/metabolismo , Probióticos/administração & dosagem , Doenças dos Suínos/tratamento farmacológico , Ração Animal/análise , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Diarreia/tratamento farmacológico , Diarreia/imunologia , Diarreia/microbiologia , Suplementos Nutricionais/análise , Microbioma Gastrointestinal , Intestinos/microbiologia , Filogenia , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia , Desmame
11.
J Vet Sci ; 20(2): e11, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30944534

RESUMO

Mammary lesions in sows can prevent suckling piglets from consuming colostrum that provides fundamental nutrients and protective immunity. Although mammary gross lesions are frequently found in sows at farms or slaughterhouses, with the exception of mastitis, they have received little research attention. In this study, we investigated mammary lesions observed in South Korean sows between 2015 and 2016. Mammary tissue samples of 82 sows showing gross lesions during meat inspection were histologically classified and immunohistochemical analysis was conducted to assess the expression of estrogen receptor (ER)-α, ER-ß, and progesterone receptor (PR) for mammary hyperplastic lesions as well as that of cluster of differentiation (CD) 3, CD79a, interleukin (IL)-1α, IL-1ß, IL-6, and IL-8 for mastitis. Furthermore, 20 swab samples were cultured, and the isolated bacteria were identified using polymerase chain reactions for 16S ribosomal RNA genes. The lesions were classified as hyperplasia, mastitis, or hyperplasia with mastitis. Immunohistochemistry results revealed that there was neither expression of ER-α nor of ER-ß, but all examined hyperplastic samples expressed PR. In addition, there was a significant correlation between CD3 and IL-1ß expressions, as well as between IL-1ß and IL-6 expressions. Regarding the identity of the isolated bacteria, Pseudomonas spp. were most frequently detected. The results of this study have revealed the incidence and characteristics of porcine mammary lesions.


Assuntos
Doenças Mamárias/veterinária , Citocinas/metabolismo , Glândulas Mamárias Animais/patologia , Receptores Estrogênicos/metabolismo , Doenças dos Suínos/patologia , Matadouros , Animais , Doenças Mamárias/metabolismo , Doenças Mamárias/microbiologia , Doenças Mamárias/patologia , Complexo CD3/metabolismo , Antígenos CD79/metabolismo , Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/metabolismo , Feminino , Interleucina-1alfa/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Glândulas Mamárias Animais/metabolismo , Glândulas Mamárias Animais/microbiologia , Mastite/metabolismo , Mastite/microbiologia , Mastite/patologia , Mastite/veterinária , Pseudomonas , Receptores de Progesterona/metabolismo , Suínos , Doenças dos Suínos/classificação , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia
12.
BMC Vet Res ; 15(1): 117, 2019 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-30992015

RESUMO

BACKGROUND: Porcine deltacoronavirus (PDCoV) is a novel coronavirus that can cause diarrhea in nursing piglets. This study was aimed to investigate the roles of host differentially expressed genes on metabolic pathways in PDCoV infections. RESULTS: Twenty thousand six hundred seventy-four differentially expressed mRNAs were identified in 5-day-old piglets responded to PDCoV experimental infections. Many of these genes were correlated to the basic metabolism, such as the peroxisome proliferator-activated receptor (PPAR) signaling pathway which plays a critical role in digestion. At the same time, in the PPAR pathway genes of fatty acid-binding protein (FABP) family members were observed with remarkably differential expressions. The differential expressed genes were associated with appetite decrease and weight loss of PDCoV- affected piglets. DISCUSSION: Fatty acid-binding protein 1 (FABP1) and fatty acid-binding protein 3 (FABP3) were found to be regulated by PDCoV. These two genes not only mediate fatty acid transportation to different cell organelles such as mitochondria, peroxisome, endoplasmic reticulum and nucleus, but also modulate fatty acid metabolism and storage as a signaling molecule outside the cell. Therefore, it can be preliminarily concluded that PPAR differential expression caused by PDCoV was mostly associated with weight loss and death from emaciation. CONCLUSIONS: The host differentially expressed genes were associated with infection response, metabolism signaling and organismal systems signaling pathways. The genes of FABP family members in the PPAR signaling pathway were the most highly altered and played important roles in metabolism. Alteration of these genes were most likely the reason of weight loss and other clinical symptoms. Our results provided new insights into the metabolic mechanisms and pathogenesis of PDCoV infection. METHODS: Animal experiment, Determination of viral growth by real-time RT-PCR, Histopathology, Immunohistochemical staining, Microarray analysis.


Assuntos
Animais Recém-Nascidos/virologia , Infecções por Coronavirus/veterinária , Coronavirus , Doenças dos Suínos/virologia , Animais , Animais Recém-Nascidos/metabolismo , Infecções por Coronavirus/metabolismo , Infecções por Coronavirus/virologia , Intestino Delgado/metabolismo , Intestino Delgado/patologia , Intestino Delgado/virologia , Jejuno/metabolismo , Jejuno/patologia , Jejuno/virologia , Redes e Vias Metabólicas/genética , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Suínos , Doenças dos Suínos/metabolismo , Transcriptoma
13.
Vet Res ; 50(1): 19, 2019 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-30836990

RESUMO

Porcine circovirus type 2 (PCV2) is an economically important swine pathogen but some extra trigger factors are required for the development of PCV2-associated diseases. By evaluating cap protein expression, viral DNA copies and the number of infected cells, the present study further confirmed that oxidative stress can promote PCV2 replication. The results showed that oxidative stress induced autophagy in PCV2-infected PK15 cells. Blocking autophagy with inhibitor 3-methyladenine or ATG5-specific siRNA significantly inhibited oxidative stress-promoted PCV2 replication. Importantly, autophagy inhibition significantly increased apoptosis in oxidative stress-treated PK15 cells. Suppression of apoptosis by benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone in conditions of autophagy inhibition restored PCV2 replication. Taken together, autophagy protected host cells against potential apoptosis and then contributed to PCV2 replication promotion caused by oxidative stress. Our findings can partly explain the pathogenic mechanism of PCV2 related to the oxidative stress-induced autophagy.


Assuntos
Apoptose , Autofagia , Infecções por Circoviridae/veterinária , Circovirus/fisiologia , Estresse Oxidativo , Doenças dos Suínos/virologia , Replicação Viral , Animais , Western Blotting/veterinária , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/metabolismo , Infecções por Circoviridae/virologia , Citocinas/metabolismo , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Glutationa/metabolismo , Peróxido de Hidrogênio/metabolismo , RNA Interferente Pequeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/metabolismo , Transfecção
14.
J Agric Food Chem ; 67(13): 3691-3701, 2019 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-30864445

RESUMO

Bile acids, synthesized in the liver and metabolized by microbiota, have emerged as important signaling molecules regulating immune responses and cell proliferation. However, the crosstalk among nutrition, microbiota, and bile acids remains unclear. Our study indicated that undernutrition in weaning piglets led to intestinal atrophy, increased colonic production, and systemic accumulation of lithocholic acid (LCA), deoxycholic acid (DCA), or their conjugated forms, which might be associated with decreased Lactobacillus abundance. Moreover, undernutrition led to increased portal fibroblast growth factor 19 ( FGF19) level, upregulated hepatic heterodimer partner ( SHP), and downregulated cholesterol 7a-hydroxylase ( CYP7A1) expression. The detrimental effects of DCA and LCA on proliferation and barrier function were confirmed in porcine enterocytes, whereas their roles in weaning piglets warrant further research. In summary, undernutrition in weaning piglets led to increased secondary bile acids production, which might be related to altered gut microbiome and enhanced farnesoid X receptor (FXR) signaling while CYP7A1 expression was suppressed.


Assuntos
Ácidos e Sais Biliares/metabolismo , Microbioma Gastrointestinal , Fígado/metabolismo , Desnutrição/veterinária , Receptores Citoplasmáticos e Nucleares/metabolismo , Doenças dos Suínos/metabolismo , Animais , Feminino , Fatores de Crescimento de Fibroblastos/genética , Fatores de Crescimento de Fibroblastos/metabolismo , Intestinos/microbiologia , Masculino , Desnutrição/genética , Desnutrição/metabolismo , Desnutrição/microbiologia , Receptores Citoplasmáticos e Nucleares/genética , Transdução de Sinais , Suínos/genética , Suínos/metabolismo , Doenças dos Suínos/genética , Doenças dos Suínos/microbiologia , Doenças dos Suínos/fisiopatologia , Desmame
15.
Vet Res ; 50(1): 17, 2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30819249

RESUMO

Porcine circovirus-associated disease (PCVAD) is one of the most serious infectious diseases in pigs worldwide. The primary causative agent of PCVAD is porcine circovirus type 2 (PCV2), which can cause lymphoid depletion and immunosuppression in pigs. Our previous study demonstrated that Laiwu (LW) pigs, a Chinese indigenous pig breed, have stronger resistance to PCV2 infection than Yorkshire × Landrace (YL) pigs. In this study, we found that the YL pigs showed more severe lymphocyte apoptosis and higher viral load in the spleen tissue than LW pigs. To illustrate the differential gene expression between healthy and infected spleens, transcriptome profiling of spleen tissues from PCV2-infected and control YL pigs was compared by RNA sequencing. A total of 90 differentially expressed genes (DEGs) was identified, including CD207, RSAD2, OAS1, OAS2, MX2, ADRB3, CXCL13, CCR1, and ADRA2C, which were significantly enriched in gene ontology (GO) terms related to the defense response to virus and cell-cell signaling, and another nine DEGs, KLF11, HGF, PTGES3, MAP3K11, XDH, CYCS, ACTC1, HSPH1, and RYR2, which were enriched in GO terms related to regulation of cell proliferation or apoptosis. Among these DEGs, the CXCL13 gene, which can suppress lymphocyte apoptosis during PCV2 infection, was significantly down-regulated in response to PCV2 infection in YL but not in LW pigs. By analysis of the regulatory elements in the promoter and 3'-untranslated region (3'-UTR) of porcine CXCL13, we found that the single nucleotide polymorphism (SNP) -1014 G (LW) > A (YL) and the Sus scrofa microRNA-296-5p (ssc-miR-296-5p) participated in regulating CXCL13 expression during the response to PCV2 infection.


Assuntos
Apoptose , Quimiocina CXCL13/metabolismo , Infecções por Circoviridae/veterinária , Circovirus , Linfócitos/virologia , Baço/virologia , Doenças dos Suínos/virologia , Animais , Western Blotting/veterinária , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/metabolismo , Infecções por Circoviridae/virologia , Circovirus/metabolismo , DNA Viral/genética , Citometria de Fluxo/veterinária , Perfilação da Expressão Gênica/veterinária , Regulação Viral da Expressão Gênica , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Análise de Sequência de RNA/veterinária , Baço/metabolismo , Baço/patologia , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/metabolismo , Carga Viral/veterinária
16.
J Anim Sci ; 97(5): 2139-2153, 2019 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-30888017

RESUMO

This study aimed to evaluate the effects of a source of dietary soluble (SF) and insoluble fiber (IF) without or with exogenous carbohydrases (xylanase, ß-glucanase, and pectinase) on diarrhea incidence, selected immune responses, and growth performance in enterotoxigenic Escherichia coli (ETEC)-challenged pigs. Sixty weaned pigs (6.9 ± 0.1 kg BW, ~23 d of age) were blocked by initial BW and placed in individual pens. Pens were randomly assigned to one of six treatments (n = 10 per treatment), including a nonchallenged control (NC), a positive challenge control (PC), the PC + a soluble fiber diet (10% sugar beet pulp) without (SF-) or with carbohydrases (SF+), and PC + an IF diet (15% corn distillers dried grains with solubles) without (IF-) or with carbohydrases (IF+). The control diet was primarily based on corn and soybean meal with 13.5% whey powder. The two sources of fiber were added at the expense of cornstarch in the control diet. Pigs were orally inoculated with 6 mL hemolytic F18 ETEC (~3.5 × 109 cfu/mL) or sham infected with 6 mL phosphate-buffered saline on day 7 (0 d postinoculation, dpi) postweaning. All ETEC challenged pigs were confirmed to be genetically susceptible to F18 ETEC. Pigs had free access to feed and water throughout the 14-d trial. Pig BW and feed intake were recorded on dpi -7, 0, and 7 or 8. Fecal swabs were collected on dpi -7, 0, 1, 2, 3, 5, and 7 or 8 to evaluate hemolytic E. coli shedding. Fecal score was visually ranked daily postchallenge to evaluate diarrhea incidence. Blood samples were collected on dpi -1, 3, and 7 or 8 at necropsy and intestinal tissues were collected at necropsy. Pigs on PC had lower dpi 1 to 7 ADG and ADFI than those on NC (P < 0.05). Compared with PC pigs, SF+ pigs had greater ADG during both pre- and postchallenge period (P < 0.05). The IF- increased postchallenge diarrhea incidence compared with PC (P < 0.05). Pigs on SF- had lower ileal E. coli attachment than PC (P < 0.05). The SF+ reduced haptoglobin and IF+ reduced C-reactive protein on dpi 3 compared with PC (P < 0.05). Compared with PC pigs, SF+ pigs tended to have lower ileal tumor necrosis factor alpha and greater ileal occludin (OCLN) mRNA (P < 0.10) and had greater (P < 0.05) colonic OCLN mRNA levels. Collectively, IF- increased incidence of diarrhea and fecal E. coli shedding compared with PC. The SF+ pigs had improved growth compared with PC pigs, likely due in part to a reduction in inflammatory intermediates.


Assuntos
Ração Animal/análise , Diarreia/veterinária , Fibras na Dieta/metabolismo , Escherichia coli Enterotoxigênica/fisiologia , Glicosídeo Hidrolases/metabolismo , Animais , Diarreia/metabolismo , Dieta/veterinária , Infecções por Escherichia coli/metabolismo , Fezes/microbiologia , Fermentação , Glicosídeo Hidrolases/genética , Íleo/metabolismo , Intestinos/microbiologia , Soja , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia , Desmame , Zea mays
17.
BMC Vet Res ; 15(1): 98, 2019 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-30909903

RESUMO

BACKGROUND: Haemophilus parasuis (HPS) is the causative agent of Glässer's disease, characterized by arthritis, fibrinous polyserositis and meningitis, and resulting in worldwide economic losses in the swine industry. Baicalin (BA), a commonly used traditional Chinese medication, has been shown to possess a series of activities, such as anti-bacterial, anti-viral, anti-tumor, anti-oxidant and anti-inflammatory activities. However, whether BA has anti-apoptotic effects following HPS infection is unclear. Here, we investigated the anti-apoptotic effects and mechanisms of BA in HPS-induced apoptosis via the protein kinase C (PKC)-mitogen-activated protein kinase (MAPK) pathway in piglet's mononuclear phagocytes (PMNP). RESULTS: Our data demonstrated that HPS could induce reactive oxygen species (ROS) production, arrest the cell cycle and promote apoptosis via the PKC-MAPK signaling pathway in PMNP. Moreover, when BA was administered, we observed a reduction in ROS production, suppression of cleavage of caspase-3 in inducing apoptosis, and inhibition of activation of the PKC-MAPK signaling pathway for down-regulating p-JNK, p-p38, p-ERK, p-PKC-α and PKC-δ in PMNP triggered by HPS. CONCLUSIONS: Our data strongly suggest that BA can reverse the apoptosis initiated by HPS through regulating the PKC-MAPK signaling pathway, which represents a promising therapeutic agent in the treatment of HPS infection.


Assuntos
Antibacterianos/uso terapêutico , Flavonoides/uso terapêutico , Haemophilus parasuis/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Monócitos/metabolismo , Doenças dos Suínos/tratamento farmacológico , Animais , Animais Recém-Nascidos/metabolismo , Animais Recém-Nascidos/microbiologia , Antibacterianos/farmacologia , Apoptose/efeitos dos fármacos , Relação Dose-Resposta a Droga , Flavonoides/farmacologia , Infecções por Haemophilus/tratamento farmacológico , Infecções por Haemophilus/metabolismo , Infecções por Haemophilus/veterinária , Monócitos/efeitos dos fármacos , Monócitos/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Suínos , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia
18.
PLoS One ; 14(3): e0212632, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30840701

RESUMO

More recently emerging strains of porcine epidemic diarrhea virus (PEDV) cause severe diarrhea and especially high mortality rates in infected piglets, leading to substantial economic loss to worldwide swine industry. These outbreaks urgently call for updated and effective PEDV vaccines. Better understanding in PEDV biology and improvement in technological platforms for virus production can immensely assist and accelerate PEDV vaccine development. In this study, we explored the ability of PEDV nucleocapsid (N) protein in improving viral yields in cell culture systems. We demonstrated that PEDV N expression positively affected both recovery of PEDV from infectious clones and PEDV propagation in cell culture. Compared to Vero E6 cells, Vero E6 cells expressing PEDV N could accelerate growth of a slow-growing PEDV strain to higher peak titers by 12 hours or enhance the yield of a vaccine candidate strain by two orders of magnitude. Interestingly, PEDV N also slightly enhances replication of porcine reproductive and respiratory virus, a PEDV relative in the Nidovirales order. These results solidify the importance of N in PEDV recovery and propagation and suggest a potentially useful consideration in designing vaccine production platforms for PEDV or closely related pathogens.


Assuntos
Proteínas do Nucleocapsídeo/biossíntese , Vírus da Diarreia Epidêmica Suína/crescimento & desenvolvimento , Animais , Infecções por Coronavirus/metabolismo , Infecções por Coronavirus/veterinária , Suínos , Doenças dos Suínos/metabolismo , Doenças dos Suínos/virologia , Células Vero
19.
Tuberculosis (Edinb) ; 114: 103-112, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30711148

RESUMO

Constraints in the characterization of microbiota community that circulates in the host have limited the extent of co-infection studies in natural populations. In this study, we used a metaproteomics approach to characterize the mandibular lymph nodes microbiota of wild boar (Sus scrofa) naturally exposed to an increasing trend of Mycobacterium tuberculosis complex (MTC) infection. Our results showed a reduction in microbiota diversity and changes in the composition, structure and functionality of the microbiota community associated with an increase in tuberculosis prevalence, from 45% in 2002/06 to 83% in 2009/12. These temporal changes were accompanied by an increase in the relative abundance of Babesia, Theileria and Pestivirus genera and a decrease in the Ascogregarina and Chlorella. A positive association was also evidenced between the prevalence of tuberculosis and the presence of microbial proteins responsible for carbohydrate transport and metabolism. Our findings suggest MTC-host-microbiota interactions at the population level, which may occur in order to ensure sufficient metabolic resources for MTC survival, growth and transmission. We strongly recommend the use of metaproteomics when studying microbiota communities in wildlife populations, for which traditional diagnostic techniques are limited and in which new organisms with a pathogenic potential for domestic animals and humans may appear.


Assuntos
Linfonodos/microbiologia , Microbiota , Doenças dos Suínos/microbiologia , Tuberculose/veterinária , Animais , Coinfecção/metabolismo , Coinfecção/microbiologia , Interações Hospedeiro-Patógeno , Mandíbula , Proteômica/métodos , Sus scrofa , Suínos , Doenças dos Suínos/metabolismo , Tuberculose/metabolismo , Tuberculose/microbiologia
20.
J Sci Food Agric ; 99(7): 3616-3627, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30628086

RESUMO

BACKGROUND: Pleurotus ostreatus mushroom (POM) is an edible mushroom with rich nutritional components and vital pharmacological properties. The present study comprised 100 cross-bred piglets, weaned at 28 days old, who were randomly assigned to four POM diets with five replicates per diet and five piglets per pen. RESULTS: POM supplementation (P < 0.05) decreased the incidence of diarrhea, and also increased the average daily feed intake and average daily gain of pigs. Fecal acetate, butyrate and propionate increased with the addition of POM. Interleukin-2, immunoglobulin G, immunoglobulin M, tumor necrosis factor-α and immunoglobulin A increased (P < 0.05) with the addition of POM. The 16S rDNA sequencing results showed that the Bacteroidetes and Firmicutes were the dominant microbial strains in the fecal samples, irrespective of POM supplementation. Shannon diversity, whole tree phylogenetic diversity, observed species and Chao1 analysis exhibited significant variation in species richness across the treatments. Principal coordinates analysis showed a significant (P < 0.1) increase in the microbial communities amongst all of the treatment groups. CONCLUSION: The results of the present study suggest that the supplementation of POM in the diet of piglets might increase feed consumption, gut microbial composition and diversity, as well as short-chain fatty acids synthesis, consequently preventing the occurrence of diarrhea and increasing the growth of piglets. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Assuntos
Ração Animal/análise , Diarreia/veterinária , Microbioma Gastrointestinal , Pleurotus/metabolismo , Doenças dos Suínos/imunologia , Suínos/crescimento & desenvolvimento , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Diarreia/imunologia , Diarreia/microbiologia , Diarreia/prevenção & controle , Dieta/veterinária , Ácidos Graxos Voláteis/metabolismo , Fezes , Feminino , Imunidade , Intestinos/imunologia , Intestinos/microbiologia , Masculino , Microbiota , Suínos/imunologia , Suínos/metabolismo , Suínos/microbiologia , Doenças dos Suínos/metabolismo , Doenças dos Suínos/microbiologia , Doenças dos Suínos/prevenção & controle
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