Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 780
Filtrar
1.
J Vet Sci ; 20(5): e50, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31565893

RESUMO

Porcine endogenous retroviruses (PERVs) integrate into germline DNA as proviral genome that enables vertical transmission from parents to their offspring. The provirus usually survives as part of the host genome rather than as an infectious agent, but may become pathogenic if it crosses species barriers. Therefore, replication-competent PERV should be controlled through selective breeding or knockout technologies. Two microRNAs (miRNAs), dual LTR1 and LTR2, were selected to inhibit the expression of PERV in primary porcine kidney cells. The inhibition efficiency of the miRNAs was compared based on their inhibition of different PERV regions, specifically long terminal repeats (LTRs), gag, pol, and env. Gene expression was quantified using real-time polymerase chain reaction and the C-type reverse transcriptase (RT) activity was determined. The messenger RNA (mRNA) expression of the PERV LTR and env regions was determined in HeLa cells co-cultured with primary porcine kidney cells. The mRNA expression of the LTR, gag, pol, and env regions of PERV was dramatically inhibited by dual miRNA from 24 to 144 h after transfection, with the highest inhibition observed for the LTR and pol regions at 120 h. Additionally, the RT activity of PERV in the co-culture experiment of porcine and human cells was reduced by 84.4% at the sixth passage. The dual LTR 1+2 miRNA efficiently silences PERV in primary porcine kidney cells.


Assuntos
Retrovirus Endógenos/fisiologia , MicroRNAs/metabolismo , Infecções por Retroviridae/veterinária , Doenças dos Suínos/genética , Animais , Linhagem Celular , Retrovirus Endógenos/genética , Rim , Infecções por Retroviridae/genética , Infecções por Retroviridae/virologia , Suínos , Doenças dos Suínos/virologia , Sequências Repetidas Terminais/fisiologia
2.
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
3.
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
4.
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
5.
Trop Anim Health Prod ; 51(6): 1307-1320, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31127494

RESUMO

Diarrhoea, a significant problem in pig rearing industry affecting pre- and post-weaning piglets is caused by enterotoxigenic Escherichia coli (ETEC). The ETEC are classified as per the fimbriae types which are responsible for bacterial attachment with enterocytes and release of toxins causing diarrhoea. However, genetic difference exists for susceptibility to ETEC infection in piglets. The different phenotypes found in pigs determine their (pigs') susceptibility or resistance towards fimbrial subtypes/variants (F4ab, F4ac, F4ad and F18). Specific receptors are present on intestinal epithelium for attachment of these fimbriae, which do not express to same level in all animals. This differential expression is genetically determined and thus their genetic causes (may be putative candidate gene or mutations) render some animals resistant or susceptible to one or more fimbrial subtypes. Genetic linkage studies have revealed the mapping location of the receptor loci for the two most frequent variants F4ab and F4ac to SSC13q41 (i.e. q arm of 13th chromosome of Sus scrofa). Some SNPs have been identified in mucin gene family, transferring receptor gene, fucosyltransferase 1 gene and swine leucocyte antigen locus that are proposed to be linked mutations for resistance/susceptibility towards ETEC diarrhoea. However, owing to the variety of fimbrial types and subtypes, it would be difficult to identify a single causative mutation and the candidate loci may involve more number of genes/regions. In this review, we focus on the genetic mutations in genes involved in imparting resistance/susceptibility to F4 or F18 ETEC diarrhoea and possibilities to use them as marker for selection against susceptible animals.


Assuntos
Escherichia coli Enterotoxigênica , Infecções por Escherichia coli/veterinária , Predisposição Genética para Doença , Doenças dos Suínos/microbiologia , Animais , Diarreia/genética , Diarreia/microbiologia , Diarreia/veterinária , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Ligação Genética , Polimorfismo de Nucleotídeo Único , Suínos , Doenças dos Suínos/genética
7.
Reprod Domest Anim ; 54(7): 972-978, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31025395

RESUMO

Follicular cysts, which is a common infertility disease, can cause financial losses in pig breeding programmes. The pathogenesis and mechanisms of the formation of follicular cysts are not understood clearly. In our previous study, the concentration of retinol-binding protein 4 (RBP-4) in the follicular fluid (FF) of the ovary with follicular cysts was found to be significantly higher than that of normal ovary, thereby suggesting that RBP-4 may be a candidate biomarker for porcine follicular cysts. To study the association of RBP-4 and follicular cysts further, we detected the polymorphisms of the RBP-4 gene and the presence of follicular cysts by PCR-Restriction fragment length polymorphism (RFLP) assay. In this study, we screened the mutations of RBP-4 gene in 79 sows with follicular cysts and 100 normal sows without cysts. Results showed that +249-63G>C polymorphisms were significantly associated with follicular cysts, and sows with CC genotype in RBP-4 gene had a high risk of developing follicular cysts. Hence, our findings further proved that RBP-4 may be a novel biomarker for follicular cysts, which may be valuable for the diagnosis of follicular cysts and molecular breeding of pigs.


Assuntos
Cisto Folicular/veterinária , Proteínas Plasmáticas de Ligação ao Retinol/genética , Doenças dos Suínos/genética , Animais , Biomarcadores , Feminino , Cisto Folicular/genética , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Sus scrofa , Suínos
8.
PLoS One ; 14(3): e0212431, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30822308

RESUMO

Swine influenza viruses (SIVs), the causal agents of swine influenza, are not only important to control due to the economic losses in the swine industry, but also can be pandemic pathogens. Vaccination is one of the most relevant strategies to control and prevent influenza infection. Current human vaccines against influenza induce strain-specific immunity and annual update is required due to the virus antigenic shift phenomena. Previously, our group has reported the use of conserved hemagglutinin peptides (HA-peptides) derived from H1-influenza virus as a potential multivalent vaccine candidate. Immunization of swine with these HA-peptides elicited antibodies that recognized and neutralized heterologous influenza viruses in vitro and demonstrated strong hemagglutination-inhibiting activity. In the present work, we cloned one HA-peptide (named NG34) into a plasmid fused with cytotoxic T lymphocyte-associated antigen (CTLA4) which is a molecule that modifies T cell activation and with an adjuvant activity interfering with the adaptive immune response. The resulting plasmid, named pCMV-CTLA4-Ig-NG34, was administered twice to animals employing a needle-free delivery approach. Two studies were carried out to test the efficacy of pCMV-CTLA4-Ig-NG34 as a potential swine influenza vaccine, one in seronegative and another in seropositive pigs against SIV. The second one was aimed to evaluate whether pCMV-CTLA4-Ig-NG34 vaccination would overcome maternally derived antibodies (MDA). After immunization, all animals were intranasally challenged with an H3N2 influenza strain. A complete elimination or significant reduction in the viral shedding was observed within the first week after the challenge in the vaccinated animals from both studies. In addition, no challenged heterologous virus load was detected in the airways of vaccinated pigs. Overall, it is suggested that the pCMV-CTLA4-Ig-NG34 vaccine formulation could potentially be used as a multivalent vaccine against influenza viruses.


Assuntos
Abatacepte , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Vírus da Influenza A Subtipo H3N2/imunologia , Vacinas contra Influenza , Infecções por Orthomyxoviridae , Peptídeos , Doenças dos Suínos , Vacinas de DNA , Eliminação de Partículas Virais , Abatacepte/genética , Abatacepte/imunologia , Abatacepte/farmacologia , Animais , Cães , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/farmacologia , Vírus da Influenza A Subtipo H3N2/genética , Vacinas contra Influenza/genética , Vacinas contra Influenza/imunologia , Vacinas contra Influenza/farmacologia , Células Madin Darby de Rim Canino , Infecções por Orthomyxoviridae/genética , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Peptídeos/genética , Peptídeos/imunologia , Peptídeos/farmacologia , Plasmídeos/genética , Plasmídeos/imunologia , Plasmídeos/farmacologia , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/imunologia , Doenças dos Suínos/prevenção & controle , Vacinação , Vacinas de DNA/genética , Vacinas de DNA/imunologia , Vacinas de DNA/farmacologia , Eliminação de Partículas Virais/efeitos dos fármacos , Eliminação de Partículas Virais/genética , Eliminação de Partículas Virais/imunologia
9.
Gene ; 701: 131-138, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30905811

RESUMO

MicroRNAs (miRNAs) play an important role in animal growth and disease development, and sequence variation in microRNAs can alter their functions. Herein, we explored the effects of mutations in the miRNA-215 precursor sequence on the miRNA-215 regulatory network and resistance to Escherichia coli (E. coli). Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to detect sequence variations in Sutai and Meishan pigs. The miR-192 precursor sequence was not mutated, but the miR-215 precursor included an AT insertion mutation at position 6 (start from the first base of the miR-215 precursor) and a C/T mutation at position 43. Wild-type (WT) and mutant miR-215 precursor expression vectors were constructed to investigate the effects of sequence variation on expression of miR-215 and its target genes DLG5 and ALCAM, cytokine levels and E. coli adhesion. Compared with the WT control group, cells harbouring the C/T mutant vector displayed reduced miR-215 expression, increased target gene expression, elevated cytokine levels and rising E. coli adhesion, whereas cells harbouring the AT insertion mutant vector were not significantly changed. The sequence variation in the miRNA-215 precursor may affect the miRNA-215 regulatory network, and alter the stability of intestinal epithelial cells (IPEC-J2 cells) and resistance to E. coli. Our findings provide guidance for future research on the regulatory mechanisms of miR-215 in porcine resistance to E. coli F18, and identifying effective genetic markers against this organism.


Assuntos
Resistência à Doença/genética , Infecções por Escherichia coli/genética , Escherichia coli , MicroRNAs/genética , Mutagênese Insercional , Polimorfismo Conformacional de Fita Simples , Doenças dos Suínos/genética , Suínos/genética , Animais , Linhagem Celular , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , MicroRNAs/imunologia , Reação em Cadeia da Polimerase , Suínos/imunologia , Suínos/microbiologia , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia
10.
Virus Genes ; 55(3): 322-331, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30919175

RESUMO

We isolated a variant of Chinese pseudorabies virus from a hunting dog with symptoms similar to Aujeszky's disease and designated the isolate MY-1 strain. The dog developed symptoms 6 days after hunting and biting a wild boar and died the day after onset. The Bam HI restriction profile of MY-1 DNA was different from those of the Japanese reference strain Yamagata-S81 and two vaccine strains, Bartha and Begonia, and resembled Bam HI-RFLP (restriction fragment length polymorphism) type IV. Complete nucleotide sequences were determined, and phylogenetic analyses revealed that MY-1 belonged to the same cluster of old Chinese strains and variant strains isolated recently in China, but most of the open reading frames of MY-1 were located on a different branch from those of these Chinese strains. Based on a gC phylogenetic analysis, MY-1 belonged to gC-genotype II composed of those Chinese strains. In mice, the 50% lethal dose (LD50) of MY-1 (103.0 TCID50) was almost the same as those of Yamagata-S81 and Bartha. The LD50 value of Begonia was 10≥4.5 TCID50. The mean survival periods of mice after infection with 104 TCID50 of MY-1, Yamagata-S81 and Bartha were 3.9 days, 2.3 days, and 8.0 days, respectively. The results suggested that the variant of Chinese PRV with slightly weaker pathogenicity than that of wild virulent viruses might be maintained in wild boars in Japan. Furthermore, we would like to propose that old Chinese strains, recent Chinese variant strains, and MY-1 should be grouped as an Asian type PRV.


Assuntos
Herpesvirus Suídeo 1/genética , Pseudorraiva/virologia , Sus scrofa/virologia , Doenças dos Suínos/virologia , Animais , Mordeduras e Picadas/veterinária , Mordeduras e Picadas/virologia , Modelos Animais de Doenças , Cães , Genótipo , Herpesvirus Suídeo 1/isolamento & purificação , Herpesvirus Suídeo 1/patogenicidade , Japão , Camundongos , Filogenia , Pseudorraiva/genética , Pseudorraiva/transmissão , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/transmissão
11.
Vet Microbiol ; 230: 235-240, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30827394

RESUMO

Pasteurella multocida is an important respiratory tract pathogen in intensive livestock farming, especially in pigs. Antimicrobial agents are frequently used to combat infections caused by this pathogen. In a study on antimicrobial resistance among respiratory tract pathogens of pigs from 30 German pig-producing farms, P. multocida isolates (n = 9) with high minimal inhibitory concentration (MIC) values of 16/304 mg/L (n = 2), 32/608 mg/L (n = 3) or ≥64/1216 mg/L (n = 4) for trimethoprim/sulfamethoxazole (1:19) and of ≥512 mg/L (n = 9) for trimethoprim (TMP) were detected in three of these farms. The genetic relatedness of the isolates was investigated via capsule-specific PCR and macrorestriction analyses with ApaI and SmaI. Pulsed-field gel electrophoresis revealed indistinguishable restriction patterns per farm, with slight differences between the three farms. All isolates represented capsular type A. Four representative isolates, that were subjected to whole genome sequencing, shared the multi-locus sequence type (ST) 3. Their plasmids were transformed into E. coli TOP10 with subsequent selection on TMP-containing agar plates. Antimicrobial susceptibility testing and plasmid analysis of the transformants confirmed that they were resistant to sulfonamides and trimethoprim and carried only a single small plasmid. This plasmid was completely sequenced and revealed a size of 6050 bp. Sequence analyses identified the presence of a resistance gene cluster comprising the genes sul2-ΔstrA-dfrA14-ΔstrA-ΔstrB. Further analysis identified a dfrA14 gene cassette being integrated into the strA reading frame. Neither the gene dfrA14 nor this gene cluster have been detected before in P. multocida.


Assuntos
Genes Bacterianos , Pasteurella multocida/genética , Suínos/microbiologia , Animais , Antibacterianos/farmacologia , Eletroforese em Gel de Campo Pulsado , Fazendas , Alemanha , Gado/microbiologia , Testes de Sensibilidade Microbiana , Família Multigênica , Pasteurella multocida/efeitos dos fármacos , Pasteurella multocida/isolamento & purificação , Plasmídeos/genética , Sulfametoxazol/farmacologia , Doenças dos Suínos/genética , Doenças dos Suínos/microbiologia , Trimetoprima/farmacologia , Sequenciamento Completo do Genoma
12.
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
13.
Arch Virol ; 164(4): 1147-1157, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30799511

RESUMO

Porcine epidemic diarrhea virus (PEDV) causes severe economic loss in the pig industry each year. To better understand the relationship between cytokines and PEDV replication, in this study, pro-inflammatory cytokine and chemokine expression profiles in Vero cells infected with PEDV were analyzed. Real-time quantitative PCR assay indicated that IL-1α, IL-1ß, TNF-α, CCL2, CCL5 and CXCL8 expression levels were significantly upregulated. Moreover, overexpression and siRNA silencing assays showed that overexpression of IL-1α, IL-1ß, TNF-α, CCL2, CCL5 and CXCL8 could significantly inhibit PEDV replication, while silencing of IL-1α, IL-1ß, TNF-α, CCL2, CCL5 and CXCL8 could significantly promote PEDV replication. Finally, a dual-luciferase reporter assay showed that nsp4 contributed to the expression of IL-1α, IL-1ß, TNF-α, CCL2, CCL5 and CXCL8 via the NF-κB pathway. Together, these data determined that PEDV nsp4 could upregulate pro-inflammatory cytokine and chemokine expression, inhibiting viral replication in vitro. These results provided novel insights for understanding the roles of cytokines in PEDV replication.


Assuntos
Quimiocinas/imunologia , Infecções por Coronavirus/veterinária , Citocinas/imunologia , Vírus da Diarreia Epidêmica Suína/fisiologia , Doenças dos Suínos/imunologia , Proteínas não Estruturais Virais/metabolismo , Replicação Viral , Animais , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Quimiocina CCL5/genética , Quimiocina CCL5/imunologia , Quimiocinas/genética , Infecções por Coronavirus/genética , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Citocinas/genética , Interações Hospedeiro-Patógeno , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-8/genética , Interleucina-8/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Vírus da Diarreia Epidêmica Suína/genética , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/virologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Células Vero , Proteínas não Estruturais Virais/genética
14.
Anim Genet ; 50(2): 162-165, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30746724

RESUMO

Pig umbilical hernia (UH) affects pig welfare and brings considerable economic loss to the pig industry. To date, the molecular mechanisms underlying pig UH are still poorly understood. To identify potential loci for susceptibility to this disease, we performed a genome-wide association study in an Erhualian × Shaziling F2 intercross population. A total of 45 animals were genotyped using Illumina Porcine SNP60 BeadChips. We observed a SNP (rs80993347) located in the calpain-9 (CAPN9) gene on Sus scrofa chromosome 14 that was significantly associated with UH (P = 1.97 × 10-10 ). Then, we identified a synonymous mutation rs321865883 (g.20164T>C) in exon 10 of the CAPN9 gene that distinguished two affected individuals (CC) from their normal full-sibs (TC). Finally, quantitative polymerase chain reaction was explored to investigate the mRNA expression profile of the CAPN9 gene in 12 tissues in Yorkshire pigs at different developmental stages (3, 90 and 180 days). CAPN9 showed high expression levels in the gastrointestinal tract at these three growth stages. The results of this study indicate that the CAPN9 gene might be implicated in UH. Further studies are required to establish a role of CAPN9 in pig UH.


Assuntos
Calpaína/genética , Estudo de Associação Genômica Ampla/veterinária , Hérnia Umbilical/veterinária , Polimorfismo de Nucleotídeo Único , Doenças dos Suínos/genética , Animais , Calpaína/metabolismo , Hérnia Umbilical/genética , Sus scrofa , Suínos
15.
J Virol ; 93(8)2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30728254

RESUMO

Identifying viral antagonists of innate immunity and determining if they contribute to pathogenesis are critical for developing effective strategies to control emerging viruses. Previously, we reported that an endoribonuclease (EndoU) encoded by murine coronavirus plays a pivotal role in evasion of host innate immune defenses in macrophages. Here, we asked if the EndoU activity of porcine epidemic diarrhea coronavirus (PEDV), which causes acute diarrhea in swine, plays a role in antagonizing the innate response in porcine epithelial cells and macrophages, the sites of viral replication. We constructed an infectious clone of PEDV-Colorado strain (icPEDV-wt) and an EndoU-mutant PEDV (icPEDV-EnUmt) by changing the codon for a catalytic histidine residue of EndoU to alanine (His226Ala). We found that both icPEDV-wt and icPEDV-EnUmt propagated efficiently in interferon (IFN)-deficient Vero cells. In contrast, the propagation of icPEDV-EnUmt was impaired in porcine epithelial cells (LLC-PK1), where we detected an early and robust transcriptional activation of type I and type III IFNs. Infection of piglets with the parental Colorado strain, icPEDV-wt, or icPEDV-EnUmt revealed that all viruses replicated in the gut and induced diarrhea; however, there was reduced viral shedding and mortality in the icPEDV-EnUmt-infected animals. These results demonstrate that EndoU activity is not required for PEDV replication in immortalized, IFN-deficient Vero cells, but is important for suppressing the IFN response in epithelial cells and macrophages, which facilitates replication, shedding, and pathogenesis in vivo We conclude that PEDV EndoU activity is a key virulence factor that suppresses both type I and type III IFN responses.IMPORTANCE Coronaviruses (CoVs) can emerge from an animal reservoir into a naive host species to cause pandemic respiratory or gastrointestinal diseases with significant mortality in humans or domestic animals. Porcine epidemic diarrhea virus (PEDV), an alphacoronavirus (alpha-CoV), infects gut epithelial cells and macrophages, inducing diarrhea and resulting in high mortality in piglets. How PEDV suppresses the innate immune response was unknown. We found that mutating a viral endoribonuclease, EndoU, results in a virus that activates both the type I interferon response and the type III interferon response in macrophages and epithelial cells. This activation of interferon resulted in limited viral replication in epithelial cell cultures and was associated with reduced virus shedding and mortality in piglets. This study reveals a role for EndoU activity as a virulence factor in PEDV infection and provides an approach for generating live-attenuated vaccine candidates for emerging coronaviruses.


Assuntos
Infecções por Coronavirus , Endorribonucleases , Interferon Tipo I/imunologia , Vírus da Diarreia Epidêmica Suína , Doenças dos Suínos , Proteínas Virais , Animais , Linhagem Celular , Infecções por Coronavirus/enzimologia , Infecções por Coronavirus/genética , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/veterinária , Endorribonucleases/genética , Endorribonucleases/imunologia , Interferon Tipo I/genética , Vírus da Diarreia Epidêmica Suína/enzimologia , Vírus da Diarreia Epidêmica Suína/genética , Vírus da Diarreia Epidêmica Suína/imunologia , Suínos , Doenças dos Suínos/enzimologia , Doenças dos Suínos/genética , Doenças dos Suínos/imunologia , Doenças dos Suínos/virologia , Proteínas Virais/genética , Proteínas Virais/imunologia , Eliminação de Partículas Virais/imunologia
16.
Virus Genes ; 55(3): 298-303, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30706196

RESUMO

Bungowannah virus, which belongs to the genus Pestivirus within the family Flaviviridae, has been associated with myocarditis and a high incidence of stillbirths in pigs. In 2003, the virus was initially detected in a large pig farming complex on two separate sites in New South Wales, Australia. Until now, it has not been detected at other locations. Despite a program of depopulation and disinfection, the virus could be only eradicated from one of the affected farm complexes, the Bungowannah unit, but became endemic on the second complex, the Corowa unit. In the present study, the genetic variability of virus isolates collected between 2003 and 2014 in the endemically infected population has been retrospectively investigated. Phylogenetic analysis carried out based on sequences of the E2 and NS5B coding regions and the full-length open-reading frame revealed that the isolates from the different farm sites are closely related, but that samples collected between 2010 and 2014 at the Corowa farm site clustered in a different branch of the phylogenetic tree. Since 2010, a high-genetic stability of this RNA virus within the Corowa farm complex, probably due to an effective adaptation of the virus to the affected pig population, could be observed.


Assuntos
Infecções por Pestivirus/genética , Pestivirus/genética , Natimorto/genética , Doenças dos Suínos/genética , Animais , Austrália , Surtos de Doenças , Pestivirus/patogenicidade , Infecções por Pestivirus/veterinária , Infecções por Pestivirus/virologia , Estudos Retrospectivos , Natimorto/veterinária , Suínos , Doenças dos Suínos/virologia
17.
Virus Genes ; 55(2): 198-208, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30712153

RESUMO

The Porcine Sapelovirus (PSV) is an enteric virus of pigs that can cause various disorders. However, there are few reports that describe the molecular characteristics of the PSV genome. In this study, almost the entire genomes of 23 PSVs detected in Japanese pigs were analyzed using bioinformatics. Analysis of the cis-active RNA elements showed that the predicted secondary structures of the internal ribosome entry site in the 5' untranslated region (UTR) and a cis-replication element in the 2C coding region were conserved among PSVs. In contrast, those at the 3' UTR were different for different PSVs; however, tertiary structures between domains were conserved across all PSVs. Phylogenetic analysis of nucleotide sequences of the complete VP1 region showed that PSVs exhibited sequence diversity; however, they could not be grouped into genotypes due to the low bootstrap support of clusters. The insertion and/or deletion patterns in the C-terminal VP1 region were not related to the topology of the VP1 tree. The 3CD phylogenetic tree was topologically different from the VP1 tree, and PSVs from the same country were clustered independently. Recombination analysis revealed that recombination events were found upstream of the P2 region and some recombination breakpoints involved insertions and/or deletions in the C-terminal VP1 region. These findings demonstrate that PSVs show genetic diversity and frequent recombination events, particularly in the region upstream of the P2 region; however, PSVs could currently not be classified into genotypes and conserved genetic structural features of the cis-active RNA elements are observed across all PSVs.


Assuntos
Diarreia/genética , Genoma Viral/genética , Infecções por Picornaviridae/virologia , Picornaviridae/genética , Animais , Diarreia/veterinária , Diarreia/virologia , Fezes/virologia , Variação Genética , Filogenia , Picornaviridae/patogenicidade , Infecções por Picornaviridae/genética , Infecções por Picornaviridae/veterinária , Suínos/genética , Suínos/virologia , Doenças dos Suínos/genética , Doenças dos Suínos/virologia
18.
Anim Genet ; 50(2): 136-142, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30724375

RESUMO

The F4ac receptor locus (F4acR), which encodes susceptibility or resistance to Escherichia coli diarrhoea, is inherited as an autosomal recessive monogenetic trait. F4acR is localized on pig chromosome 13 (SSC13q41-q44) near the MUC13 gene. Two flanking markers (CHCF1 and ALGA0106330) with a high linkage disequilibrium (LD) with F4acR were found to be effective for the genetic identification of F4ac-resistant pigs in the Swiss Large White breed (one recombinant out of 2034 genotyped pigs). Three recombinant boars, one each from the Duroc, Swiss Landrace and Piétrain breeds, were genotyped with seven different markers and phenotyped by means of a microscopic adhesion test. Only ALGA0072075, CHCF1 and CHCF3 indicated the correct phenotype. To test the effect of the resistance allele on production traits, 530 Large White pigs from the national test station were investigated. A significant difference existed among the F4acR locus genotypes in the intramuscular fat content of the longissimus dorsi muscle, whereas no other production traits were influenced by the resistance allele. The frequency of the CHCF1-C and ALGA0106330-A alleles associated with resistance in the Swiss Large White population was 60%, which is advantageous for implementing this trait in a breeding programme to select for E. coli F4ac-resistant animals. The selection of resistant pigs should start on the male side due to the inability of resistant sows to produce sufficient amounts of protecting antibodies in the colostrum. Selection of genetically F4ac-resistant pigs is a sustainable and suitable alternative to decreasing animal loss and antibiotic use due to diarrhoea.


Assuntos
Aderência Bacteriana , Diarreia/veterinária , Infecções por Escherichia coli/veterinária , Marcadores Genéticos , Desequilíbrio de Ligação , Doenças dos Suínos/genética , Animais , Diarreia/genética , Diarreia/microbiologia , Escherichia coli/fisiologia , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Feminino , Genótipo , Masculino , Sus scrofa , Suínos , Doenças dos Suínos/microbiologia
19.
Theriogenology ; 127: 49-55, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30665073

RESUMO

Cryptorchidism, a condition of one or two undescended testicles, is a common congenital disease in pigs, causing loss in the pig industry. One of the major factors affecting testicular descent is the androgen receptor (AR), which binds to androgen and then regulates the expression of androgen-responsive genes in the inguinoscrotal phase of testicular descent. AR expression has been reported to regulate apoptosis in testicular stem cells. The present study aimed to immunohistochemically examine AR and Ki-67 protein expression and apoptosis detection in unilateral undescended testicles (UDT) and descended testicles in cryptorchid pigs (DT) of suckling (aged 1-2 weeks), nursery (aged 6 weeks) and growing-finishing pigs (aged 12, 15 and 20 weeks) and in normal testicles (NT) at 1-2 and 12 weeks of age. At 1-2 weeks, decreased expression of AR was observed in UDT and DT compared with NT and was lower than that at 6-20 weeks. The expression of Ki-67, a marker of cell proliferation, in UDT and DT at 12 weeks was lower than that in NT at the same age. In addition, Ki-67 expression in UDT at 6 and 12 weeks was lower than that in UDT at 1-2 and 15-20 weeks. More testicular apoptosis was revealed in UDT at 1-2 weeks than in DT and NT at the same age. At 15-20 weeks, more apoptosis was detected in UDT than in DT. Positive correlation of AR expression in DT at 6 and 12 weeks was also noted, in addition to the association of the expression of AR and Ki-67 in NT at 12 weeks. Taken together, this study unveiled the low expression of AR and high apoptosis detection in UDT, whereas low expression of AR and low apoptosis detection were noted in DT in suckling piglets. Diminished cell proliferation was shown in UDT at 6-12 weeks, whereas high apoptosis was observed in UDT at 15-20 weeks. High expression of AR was shown only in nursery pigs. Distinct expression of AR in DT and NT at 1-2 and 12 weeks indicated that both conditions were not interchangeable.


Assuntos
Apoptose , Proliferação de Células , Criptorquidismo/veterinária , Receptores Androgênicos/metabolismo , Doenças dos Suínos/metabolismo , Animais , Criptorquidismo/genética , Criptorquidismo/metabolismo , Criptorquidismo/patologia , Imuno-Histoquímica , Masculino , Receptores Androgênicos/genética , Maturidade Sexual , Suínos , Doenças dos Suínos/genética , Doenças dos Suínos/patologia , Testículo/metabolismo
20.
PLoS Genet ; 15(1): e1007759, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30699111

RESUMO

Balancing selection provides a plausible explanation for the maintenance of deleterious alleles at moderate frequency in livestock, including lethal recessives exhibiting heterozygous advantage in carriers. In the current study, a leg weakness syndrome causing mortality of piglets in a commercial line showed monogenic recessive inheritance, and a region on chromosome 15 associated with the syndrome was identified by homozygosity mapping. Whole genome resequencing of cases and controls identified a mutation causing a premature stop codon within exon 3 of the porcine Myostatin (MSTN) gene, similar to those causing a double-muscling phenotype observed in several mammalian species. The MSTN mutation was in Hardy-Weinberg equilibrium in the population at birth, but significantly distorted amongst animals still in the herd at 110 kg, due to an absence of homozygous mutant genotypes. In heterozygous form, the MSTN mutation was associated with a major increase in muscle depth and decrease in fat depth, suggesting that the deleterious allele was maintained at moderate frequency due to heterozygous advantage (allele frequency, q = 0.22). Knockout of the porcine MSTN by gene editing has previously been linked to problems of low piglet survival and lameness. This MSTN mutation is an example of putative balancing selection in livestock, providing a plausible explanation for the lack of disrupting MSTN mutations in pigs despite many generations of selection for lean growth.


Assuntos
Músculo Esquelético/fisiopatologia , Miostatina/genética , Seleção Genética , Doenças dos Suínos/genética , Alelos , Animais , Códon sem Sentido/genética , Pé/fisiopatologia , Heterozigoto , Homozigoto , Mutação , Fenótipo , Sus scrofa/genética , Suínos , Doenças dos Suínos/fisiopatologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA