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1.
Arch Virol ; 169(9): 179, 2024 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-39150476

RESUMO

Seven novel porcine parvoviruses (PPV2 to PPV8) have been discovered in the last two decades. The last one reported was PPV8 in China in 2022, which was proposed to be a member of the genus Protoparvovirus. Here, we report the first detection of PPV8 outside China - in two provinces from Colombia. Six out of 146 (4.1%) pigs showing porcine respiratory disease (PRD) tested positive for PPV8. Sequencing and phylogenetic analysis of two Colombian PPV8 isolates (GenBank database accession numbers PP335559 and PP335560) showed them to be members of the genus Protoparvovirus. Furthermore, PPV8 was detected in coinfections with porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), which are associated with PRD.


Assuntos
Infecções por Parvoviridae , Parvovirus Suíno , Doenças dos Suínos , Animais , Coinfecção/virologia , Coinfecção/veterinária , Coinfecção/epidemiologia , Colômbia/epidemiologia , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/virologia , Infecções por Parvoviridae/epidemiologia , Parvovirus Suíno/genética , Parvovirus Suíno/isolamento & purificação , Parvovirus Suíno/classificação , Filogenia , Síndrome Respiratória e Reprodutiva Suína/virologia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , Suínos , Doenças dos Suínos/virologia , Doenças dos Suínos/epidemiologia
2.
Braz J Microbiol ; 55(1): 1017-1022, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38041718

RESUMO

Porcine reproductive and respiratory syndrome (PRRS) and African swine fever (ASF) are economically important diseases of pigs throughout the world. During an outbreak, all age groups of animals except piglets < 1 month of age were affected with symptoms of high fever, cutaneous hemorrhages, vomition with blood, diarrhea, poor appetite, ataxia, and death. The outbreak was confirmed by the detection of the N gene of the porcine reproductive and respiratory syndrome virus (PRRSV) and the VP72 gene of the African swine fever virus (ASFV) by PCR in representative blood samples from affected pigs followed by Sanger sequencing. Mixed infection was also confirmed by simultaneous detection of both the viruses using multiplex PCR. Phylogenetic analysis of both the viruses revealed that the outbreak was related to ASFV and PRRSV strains from China which were also closely related to the PRRSV and ASFV strains from the recent outbreak from India. The study confirmed the involvement of genotype II of ASFV and genotype 2 of PRRSV in the present outbreak. Interestingly, PRRSV associated with the present outbreak was characterized as a highly pathogenic PRRSV. Therefore, the present study indicates the possibility of future waves or further outbreaks of these diseases (PRRS and ASF) in this region. This is the first report of ASFV and PRRSV co-infection in pigs from India.


Assuntos
Vírus da Febre Suína Africana , Febre Suína Africana , Coinfecção , Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Suínos , Animais , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Febre Suína Africana/genética , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Febre Suína Africana/epidemiologia , Coinfecção/epidemiologia , Coinfecção/veterinária , Filogenia
3.
Prev Vet Med ; 220: 106032, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37778218

RESUMO

Despite the economic importance of PRRS and its high prevalence in Costa Rica, there are no studies on the bioeconomic impact of the disease in the country or, even, in Central America. Such studies are essential in finding cost-effective preventive measures tailored for different production circumstances. Therefore, the objective of this study was to evaluate economic and production parameters of a PRRSV-infection for a medium-sized farrow-to-finish pig farm system in Costa Rica with a farm-level stochastic Monte Carlo simulation model. The effect of PRRS was assessed by scenario analysis, in which a baseline PRRS-free situation was compared against three alternative scenarios that assumed low, medium and high PRRS effects. The PRRS effects were based on data from local farms, scientific literature and expert opinion. Sensitivity analyses were performed to assess the impact of key input parameters on output variables. Results show that at the animal level, changes between the baseline and the PRRS-high scenario were estimated as: + 25 d in age to slaughter, - 9.9 pigs to slaughter (per breeding sow/yr), + 6% annual replacement rate, - 255 d in sow productive lifetime, - 6.9 mo in age at culling of sows, and + 24 non- productive days. For a medium size local farm (n = 588 sows), a reduction of 5826 fat pigs to slaughter per farm/yr from baseline compared to PRRS-high scenario was observed. PRRS-induced loss per farm per year was estimated at -US $142,542, US $180,109 and -US $524,719 for PRRS-low, medium and high scenarios, respectively. Revenues/costs ratio changed from 1.12 in the baseline to 0.89 in the PRRS-high scenario. The production cost per kg carcass weight increased from US $2.63 for the baseline to US $3.35 in the PRRS-high scenario. PRRS-induced loss was estimated at US $77.1 per slaughtered pig/yr and US $892 per breeding sow/yr for the PRRS-high scenario. Results from the model indicate that pig farms with medium to high prevalence of PRRS will require optimal market conditions in order to have positive economic outcomes. These results can be helpful in the design of better control strategies for PRRS.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Suínos , Animais , Feminino , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Fazendas , Costa Rica , Criação de Animais Domésticos/métodos
4.
Viruses ; 16(1)2023 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-38275949

RESUMO

We analyzed the T-cell responses induced by lineal epitopes of glycoprotein 5 (GP5) from PRRSV to explore the role of this protein in the immunological protection mediated by T-cells. The GP5 peptides were conjugated with a carrier protein for primary immunization and booster doses. Twenty-one-day-old pigs were allocated into four groups (seven pigs per group): control (PBS), vehicle (carrier), PTC1, and PTC2. Cytokine levels were measured at 2 days post-immunization (DPI) from serum samples. Cytotoxic T-lymphocytes (CTLs, CD8+) from peripheral blood were quantified via flow cytometry at 42 DPI. The cytotoxicity was evaluated by co-culturing primed lymphocytes with PRRSV derived from an infectious clone. The PTC2 peptide increased the serum concentrations of pro-inflammatory cytokines (i.e., TNF-α, IL-1ß, IL-8) and cytokines that activate the adaptive cellular immunity associated with T-lymphocytes (i.e., IL-4, IL-6, IL-10, and IL-12). The concentration of CTLs (CD8+) was significantly higher in groups immunized with the peptides, which suggests a proliferative response in this cell population. Primed CTLs from immunized pigs showed cytolytic activity in PRRSV-infected cells in vitro. PTC1 and PTC2 peptides induced a protective T-cell-mediated response in pigs immunized against PRRSV, due to the presence of T epitopes in their sequences.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Vacinas Virais , Suínos , Animais , Vírus da Síndrome Respiratória e Reprodutiva Suína/metabolismo , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Anticorpos Antivirais , Citocinas/metabolismo , Fator de Necrose Tumoral alfa , Epitopos
5.
Sci Rep ; 12(1): 21595, 2022 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-36517668

RESUMO

This research aimed to identify bioactive compounds from Caesalpinia sappan extract that function as novel porcine reproductive and respiratory syndrome virus (PRRSV) infection inhibitors by computational molecular screening. We obtained a set of small-molecule compounds predicted to target the scavenger receptor cysteine-rich domain 5 (SRCR5) of CD163. In addition, the functions of positive hits were assessed and verified utilizing an in vitro antiviral activity assay with PRRSV-infected MARC-145 cells. Combining molecular docking with the results of binding affinity and ligand conformation, it was found that brazilin had the highest binding energy with the SRCR5 receptor compared to catechin and epicatechin (- 5.8, - 5.5, and - 5.1 kcal/mol, respectively). In terms of molecular mechanics, the binding free energy between the SRCR5 receptor was - 15.71 kcal/mol based on the Poisson-Boltzmann surface area of brazilin. In addition, PRRSV infection in MARC-145 cells was significantly inhibited by brazilin compared to the control (virus titer, 4.10 vs. 9.25 TCID50/mL, respectively). Moreover, brazilin successfully limited the number of PRRSV RNA copies in MARC-145 cells as determined by RT-qPCR. By inhibiting the PRRSV-CD163 interaction with brazilin from Caesalpinia sappan, it may be possible to prevent PRRSV infection in pigs, as suggested by this research.


Assuntos
Caesalpinia , Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Viroses , Suínos , Animais , Caesalpinia/química , Síndrome Respiratória e Reprodutiva Suína/tratamento farmacológico , Simulação de Acoplamento Molecular
6.
Arch Microbiol ; 204(7): 367, 2022 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-35661262

RESUMO

Data regarding PRRSV-2 in South America are scant and a coordinated criterion for molecular characterization is needed. A phylogenetic analysis was performed using a dataset of 76 ORF5 sequences from South America, and results showed the identification of lineage 5 in the early 2000s and the predominance of lineage 1 at least since 2013. Lineage 1 sequences were further classified into sub-lineages according to a recent molecular characterization study of PRRSV-2 in United States. Our results revealed the recent identification in Uruguay of PRRSV-2 ORF5 sequences of lineage 1 sub-lineage C. Two additional sub-lineages were identified in South America, 1G in Chile and 1A in Peru. Continuous updating the molecular epidemiology of circulating viruses with coordinated investigations among countries is required to control and prevent the emergence of genetic variants of PRRSV-2.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Animais , Chile/epidemiologia , Variação Genética , Epidemiologia Molecular , Filogenia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Suínos , Estados Unidos
7.
Vet Med Sci ; 8(2): 610-618, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35023299

RESUMO

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped RNA virus in the order Nidovirales, family Arteriviridae, genus Betaarterivirus. Antibodies against nonstructural proteins (NSPs) from this virus can be found in pigs starting 4 days postinfection and they remain detectable for several months. OBJECTIVE: The goal of this study was to evaluate the immunogenicity and antigenic properties of recombinant proteins NSP1 and NSP11 expressed in Escherichia coli cells, as well as to assess the neutralization activity that they elicit. METHODS: We obtained the complete ORF-1 genes coding for NSP1 and NSP11 from PRRSV using the VR-2332 strain. Cloning was performed with the pET23a(+) vector with a histidine tag (His6), linearized by restriction enzyme digestion; the expression of the NSP1 and NSP11 clones was induced in OverExpress C41(DE3) chemically competent cells. Recombinant proteins were used to generate hyperimmune sera and we perform serological assays to confirm neutralizing antibodies. RESULTS: The expressed recombinant NSP1 and NSP11 were found to be immunogenic when injected in pigs, as well as demonstrated higher specificity in recognition of antigen in field sera from pigs positive infected with PRRSV. Furthermore, both NSP1 and NSP11 recombinant proteins elicited PRRSV neutralizing antibodies. CONCLUSIONS: In this study, we demonstrated the immune humoral response to NSP 1 and NSP11, and neutralizing-antibody response to PRRSV VR2332 strain in sera from hyperimmunized pigs.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Doenças dos Suínos , Animais , Anticorpos Neutralizantes , Formação de Anticorpos , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Proteínas Recombinantes/genética , Suínos , Proteínas não Estruturais Virais/química
8.
Transbound Emerg Dis ; 69(2): 903-907, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33590723

RESUMO

Porcine reproductive and respiratory syndrome (PRRS) is a viral disease characterized by reproductive impairment or failure in breeding animals, and a respiratory disease in pigs of any age. Brazil is the fourth largest pork producer and exporter globally, and PRRS virus (PRRSV) infection has never been reported in the country. This study aimed to investigate the status of porcine biological samples from commercial swine herds, quarantined imported boars, wild boars and feral pigs to update PRRS information in Brazil. A total of 14,382 samples were collected from 2008 to 2020, including sera (n = 12,841), plasma (n = 1,000) and oral fluids (n = 541), comprehending 137 herds and free-living pigs in eight Brazilian states. One out of 1,000 (0.1%) plasma and 15 out of 12,841 (0.11%) serum samples tested positive for PRRSV antibodies through ELISA. Upon ELISA retesting, only the plasma sample, from one 8-day-old piglet remained positive. All sixteen previously PRRSV antibody-positive samples were tested through RT-PCR and found to be negative. The presence of false-positive or singleton reactors are quite expected. Thus, the use of different/alternative diagnostic tests is indicated for an efficient PRRSV detection. Taken together, our findings demonstrated no conclusive evidence of PRRSV infection in the tested pigs, highlighting the importance to reinforce the surveillance program to prevent the introduction and eventual dissemination of PRRSV in Brazil.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Doenças dos Suínos , Animais , Anticorpos Antivirais , Brasil/epidemiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Estudos Retrospectivos , Suínos
9.
Front Immunol ; 12: 712109, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34394113

RESUMO

Dendritic cells (DCs) are the most potent antigen-presenting cells, unique to initiate and coordinate the adaptive immune response. In pigs, conventional DCs (cDCs), plasmacytoid DCs (pDCs), and monocyte-derived DCs (moDCs) have been described in blood and tissues. Different pathogens, such as viruses, could infect these cells, and in some cases, compromise their response. The understanding of the interaction between DCs and viruses is critical to comprehend viral immunopathological responses. Porcine reproductive and respiratory syndrome virus (PRRSV) is the most important respiratory pathogen in the global pig population. Different reports support the notion that PRRSV modulates pig immune response in addition to their genetic and antigenic variability. The interaction of PRRSV with DCs is a mostly unexplored area with conflicting results and lots of uncertainties. Among the scarce certainties, cDCs and pDCs are refractory to PRRSV infection in contrast to moDCs. Additionally, response of DCs to PRRSV can be different depending on the type of DCs and maybe is related to the virulence of the viral isolate. The precise impact of this virus-DC interaction upon the development of the specific immune response is not fully elucidated. The present review briefly summarizes and discusses the previous studies on the interaction of in vitro derived bone marrow (bm)- and moDCs, and in vivo isolated cDCs, pDCs, and moDCs with PRRSV1 and 2.


Assuntos
Células Dendríticas/imunologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Animais , Apresentação de Antígeno , Antígenos Virais/imunologia , Medula Óssea , Células Dendríticas/classificação , Previsões , Monócitos , Vírus da Síndrome Respiratória e Reprodutiva Suína/patogenicidade , Suínos , Linfócitos T Reguladores/imunologia , Vacinas Virais , Virulência
10.
Trop Anim Health Prod ; 53(4): 441, 2021 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-34406521

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant economic losses to the swine industry worldwide. Little is known regarding the epidemiology of this infection in tropical countries. To address this problem in Costa Rica, a seroepidemiological study was carried out in two phases. In the first phase, a pilot study was conducted in nine farms with the clinical diagnosis of PRRSV. In total, 265 pig serum samples were collected from animals ranging in age from 1 to 15 weeks of age. This study aimed to establish the duration of maternal immunity in piglets, to identify the period of viremia, and to determine when seroconversion occurs. In the second phase, a cross-sectional serology study was performed on a representative sample of the Costa Rican national herds in the second phase. The twenty-five selected farms represent all provinces and were classified according to herd size (100 to 2000 sows). In each farm, pigs aged 8, 10, and 12 weeks were sampled, as well as gilts based on the pilot study. In total 1281 pigs were sampled across all 25 farms. The aim of the cross-sectional study was to quantify the seroprevalence of PRRSV in Costa Rican pig farms and to describe its geographical distribution in this tropical country. The prevalence of positive farms was 44% (11/25), and these farms were located in six of the seven provinces of Costa Rica. Overall, 58% (344/596) of the pigs were seropositive to PRRSV. The age of the pigs and the ecozone where farms were located were significantly related with PRRSV seroprevalence in animals and herds, respectively.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Doenças dos Suínos , Animais , Anticorpos Antivirais , Costa Rica/epidemiologia , Estudos Transversais , Fazendas , Feminino , Projetos Piloto , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Estudos Soroepidemiológicos , Suínos
11.
BMC Vet Res ; 17(1): 217, 2021 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-34118903

RESUMO

BACKGROUND: Worldwide, Porcine Reproductive and Respiratory Syndrome (PRRS) is among the diseases that cause the highest economic impact in modern pig production. PRRS was first detected in Costa Rica in 1996 and has since then severely affected the local swine industry. Studies of the molecular characterization of circulating strains, correlation with clinical records, and associations with pathogens associated with Porcine Respiratory Disease Complex (PRDC) have not been done in Costa Rica. RESULTS: Sequencing and phylogenetic analysis of ORF5 proved that PRRSV-2 was the only species detected in all locations analyzed. These sequences were grouped into three clusters. When comparing samples from San Jose, Alejuela, and Puntarenas to historical isolates of the previously described lineages (1 to 9), it has been shown that these were closely related to each other and belonged to Lineage 5, along with the samples from Heredia. Intriguingly, samples from Cartago clustered in a separate clade, phylogenetically related to Lineage 1. Epitope analysis conducted on the GP5 sequence of field isolates from Costa Rica revealed seven peptides with at least 80% amino acid sequence identity with previously described and experimentally validated immunogenic regions. Previously described epitopes A, B, and C, were detected in the Santa Barbara-Heredia isolate. CONCLUSIONS: Our data suggest that the virus has three distinct origins or introductions to the country. Future studies will elucidate how recently introduced vaccines will shape the evolutionary change of circulating field strains.


Assuntos
Fases de Leitura Aberta/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Sequência de Aminoácidos , Animais , Costa Rica/epidemiologia , Epitopos/análise , Filogenia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Suínos
12.
Trop Anim Health Prod ; 53(1): 75, 2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33404929

RESUMO

Pork accounts for almost one-third of the meat consumed worldwide. Infectious diseases have a marked impact on pig production. Epidemiological indicators are considered the most useful criteria in decision-making; however, a health status assessment remains a challenge at the national and regional levels. This study proposes a health index including herd-losses, morbidity, fatality, and type of diseases, to rate the health situation in a region or country; it contributes to assessing the effectiveness of control, damage manifestation, and trends. It is a multidimensional index with a structure of triads and simple quantitative, semi-quantitative, and qualitative expressions that use flexible and dynamics limits. With it, we analyzed twenty-one countries in 2005-2018, focusing on African swine fever, classical swine fever, foot-mouth-disease, and porcine respiratory and reproductive syndrome, diseases that caused 72% of the morbidity. Our multidimensional approach estimates farm, local, and regional impact from infectious agents and outbreaks, and apprises trends aiming to be useful to control measures, strategic actions, and animal health policies.


Assuntos
Febre Suína Africana/epidemiologia , Peste Suína Clássica/epidemiologia , Monitoramento Epidemiológico/veterinária , Febre Aftosa/epidemiologia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Febre Suína Africana/mortalidade , Febre Suína Africana/virologia , Animais , Peste Suína Clássica/mortalidade , Peste Suína Clássica/virologia , Febre Aftosa/mortalidade , Febre Aftosa/virologia , Síndrome Respiratória e Reprodutiva Suína/mortalidade , Síndrome Respiratória e Reprodutiva Suína/virologia , Sus scrofa , Suínos
13.
Vet Microbiol ; 248: 108833, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32891948

RESUMO

Currently live attenuated porcine reproductive and respiratory syndrome (PRRS) and classical swine fever (CSF) vaccines are widely used in Chinese swine herds. However, the mutual effects of vaccination procedures and severe stress caused by successive vaccinations harm piglets and make it difficult to stimulate robust and effective immune responses. In our previous study, a recombinant PRRS virus (PRRSV) vectored vaccine candidate rPRRSV-E2, which expresses CSF virus (CSFV) E2 protein, has been demonstrated being able to protect piglets against lethal challenge of highly-pathogenic (HP)-PRRSV and CSFV. In this study, we determine whether preexisting maternally derived antibodies (MDA) interfere with the immune efficacy of rPRRSV-E2. 8 experimental groups of piglets, with or without PRRSV MDAs or CSFV MDAs were immunized with a single dose of 105 TCID50 rPRRSV-E2 or DMEM and challenged with HP-PRRSV or CSFV. Clinical characteristics, PRRSV- or CSFV-specific antibodies, viremia and pathological changes were monitored, examined and analyzed. The results showed that rPRRSV-E2-vaccinated piglets, either with or without MDAs directed against PRRSV or CSFV were completely protected from the lethal challenge of HP-PRRSV or CSFV. These results demonstrate that the MDAs do not interfere with the immune efficacy of rPRRSV-E2, which indicates that rPRRSV-E2 could have great significance in the effective prevention and control of HP-PRRSV and CSFV.


Assuntos
Anticorpos Antivirais/imunologia , Imunidade Materno-Adquirida , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Vacinas Virais/imunologia , Fatores Etários , Animais , Feminino , Vetores Genéticos , Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Suínos , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologia
14.
J Am Soc Mass Spectrom ; 31(6): 1302-1312, 2020 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-32379441

RESUMO

Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease characterized by severe reproductive failure in sows, acute respiratory disorders in growing pigs, and high mortality in piglets. The causative agent of this syndrome is the PRRS virus (PRRSV), an RNA virus belonging to the Arteriviridae family. To date, several quantitative approaches of proteomics have been applied to analyze the gene expression profiles during PRRSV infection in PAMs and MARC-145 cells, and few proteins have been consistent among independent studies, probably due to the differences in the levels of virulence of different PRRSV strains used and/or due to analytical conditions. In this study, total proteins isolated from noninfected and infected MARC-145 cells with a Mexican PRRSV strain were relatively quantified using label-free based DIA approach in combination with ion-mobility separation. As a result, 1456 quantified proteins were found to be shared between the control and infected samples. Afterward, these proteins were filtered, and 699 of them were considered without change. Also, 17 proteins were up-regulated and 19 proteins were down-regulated during the PRSSV infection. Bioinformatic analysis revealed that many of the differentially expressed proteins are involved in processes like antigen processing, presentation of antigens, response to viruses, response to IFNs, and innate immune response, among others. The present work is the first one which provides a detailed proteomic analysis through label-free based DIA approach in MARC-145 cells during the infection with a Mexican PRRSV strain.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Proteoma , Proteômica/métodos , Animais , Linhagem Celular , Chlorocebus aethiops , Interações Hospedeiro-Patógeno , Espectrometria de Massas/métodos , Síndrome Respiratória e Reprodutiva Suína/metabolismo , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína , Mapas de Interação de Proteínas , Proteoma/análise , Proteoma/química , Proteoma/metabolismo , Suínos
15.
Viruses ; 11(11)2019 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-31683569

RESUMO

Conventional dendritic cells (cDCs) cannot be infected by porcine reproductive and respiratory syndrome virus (PRRSV) but respond to infection via cytokine production, indicating a possible role in initiation/regulation of the immune response against PRRSV. In this work, we evaluated the responses of splenic and blood cDCs, with DEC205+CADM1+CD172a+/- phenotype, as well as those of CD163+ cells against PRRSV and porcine epidemic diarrhea virus (PEDV). Both populations were incubated in the presence of PRRSV or PEDV with and without naïve CD3+ T cells, and cytokine responses were evaluated by qPCR and ELISA. Our results showed that cDCs, but not CD163+ cells, produced IL-12 in response to PRRSV. PEDV did not induce IL-12 production. Cocultures of cDCs and autologous naïve CD3+ cells resulted in decreased IL-12 production and low expression of IFN-γ transcripts in response to PRRSV. Interestingly, cDCs increased the proliferation of naïve T cells in the presence of PRRSV compared with that achieved with monocytes and peripheral blood mononuclear cells (PBMCs). Cocultures of CD163+ cells induced IL-10 and IL-4 expression in the presence of PRRSV and PEDV, respectively. In conclusion, cDCs can selectively produce IL-12 in response to PRRSV but poorly participate in the activation of naïve T cells.


Assuntos
Infecções por Coronavirus/veterinária , Células Dendríticas/imunologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Linfócitos T , Animais , Antígenos CD/sangue , Antígenos de Diferenciação Mielomonocítica/sangue , Molécula 1 de Adesão Celular/sangue , Infecções por Coronavirus/imunologia , Citocinas/sangue , Células Dendríticas/virologia , Interleucina-10/sangue , Interleucina-12/sangue , Interleucina-4/sangue , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/virologia , Monócitos/imunologia , Monócitos/virologia , Vírus da Diarreia Epidêmica Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Cultura Primária de Células , Receptores de Superfície Celular/sangue , Baço/citologia , Baço/imunologia , Baço/virologia , Suínos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/virologia , Linfócitos T/imunologia , Linfócitos T/virologia
16.
Transbound Emerg Dis ; 66(3): 1107-1113, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30688036

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant economic losses to the swine industry worldwide. While PRRSV has been endemic in North America since 1989, it was not until 1999 that the virus was first described in South America. Notably, recently an increased number of PRRSV outbreaks have been reported in South American countries. However, epidemiological information related to these outbreaks is limited and the genetic characteristics of the PRRSV strains circulating in the region are poorly understood. In this study, we describe the genetic analyses of PRRSV strains associated with severe PRRS outbreaks in Peru. Samples originating from 14 farms located in two Departments in Peru (Lima and Arequipa), were subjected to RT-PCR amplification of the PRRSV ORF5 gene and sequencing followed by restriction fragment length polymorphism (RFLP) analysis. Results demonstrated the circulation of PRRSV-2 in Peru. Notably ORF5 RFLP typing revealed that 15 (75%) of the PRRSV strains detected in this study belong to the RFLP 1-7-4 type. Phylogenetic analysis showed that the Peruvian strains are closely related to the highly virulent PRRSV 1-7-4 strains that emerged in the US in 2013-2014. Results here indicate the presence of highly virulent PRRSV 1-7-4 strains in Peru and provide important information on the geographical distribution of PRRSV, confirming the recent geographical expansion of this important swine pathogen towards South America.


Assuntos
Surtos de Doenças/veterinária , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Animais , Fazendas , Feminino , Geografia , Masculino , Peru/epidemiologia , Filogenia , Polimorfismo de Fragmento de Restrição , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/patogenicidade , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Suínos , Proteínas do Envelope Viral/genética
17.
Virology ; 529: 65-72, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30665099

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) infects monocyte-derived DCs, and previous reports have shown that PRRSV does not infect conventional DCs (cDCs) in vitro, but the effects on cDCs from lymphoid tissues are unknown. This study analyzed the response and susceptibility of tonsil DEC205+cDCs from infected pigs. We confirmed the phenotype and lineage of bona fide tonsil cDCs with the mRNA expression of FLT3+ and the phenotype MHCII+CADM1highDEC205+ (DEC205+cDCs). These cells were not infected by PRRSV, whereas CD163+ tonsil cells were infected. The numbers of tonsil cDCs and CD163+ cells were not affected by PRRSV, in contrast to the reduction in alveolar macrophage numbers. DEC205+cDCs exhibited an increase in the expression of IL-12 at 5 days postinfection, suggesting a proinflammatory response by these cells to the virus. In summary, this study confirms that, in vitro and in vivo, cDCs are not susceptible to PRRSV but can respond against it.


Assuntos
Células Dendríticas/virologia , Tonsila Palatina/citologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Animais , Antígenos CD/genética , Antígenos CD/metabolismo , Molécula 1 de Adesão Celular/genética , Molécula 1 de Adesão Celular/metabolismo , Regulação da Expressão Gênica/imunologia , Interleucina-12/genética , Interleucina-12/metabolismo , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Complexo Principal de Histocompatibilidade , Antígenos de Histocompatibilidade Menor/genética , Antígenos de Histocompatibilidade Menor/metabolismo , Síndrome Respiratória e Reprodutiva Suína/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Suínos , Tirosina Quinase 3 Semelhante a fms/genética , Tirosina Quinase 3 Semelhante a fms/metabolismo
18.
Prev Vet Med ; 160: 116-122, 2018 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-30388993

RESUMO

Biosecurity is defined as the set of practices carried out to prevent the introduction and spread of infectious agents in a herd. These practices are essential in swine production, especially for highly infectious agents such as porcine reproductive and respiratory syndrome virus (PRRSv). Even with years of research and experience over the last three decades, PRRSv is still causing productivity losses and is the major health problem affecting the global swine industry. Despite knowledge of the various ways in which the virus can be transmitted from one herd to another (e.g. animals, semen, truck, air, and people), determining the most frequent ways in which the virus is transmitted in the field is difficult. A systematic approach to assess vulnerabilities at a herd level related to PRRSv transmission could help producers prioritize biosecurity practices to reduce or avoid the occurrence of outbreaks. The aim of this study was to develop a biosecurity vulnerability score that represents the relative vulnerability of swine breeding herds to the introduction of PRRSv. To create the biosecurity vulnerability score (outcome), a multi-criteria decision analysis methodology was used to rank and quantify biosecurity practices based on expert opinion. To validate the biosecurity vulnerability score, a survey of biosecurity practices and PRRS outbreak histories in 125 breed-to-wean herds in the U.S. swine industry was used. Data on the frequency of PRRS outbreaks was used to test the hypothesis that biosecurity vulnerability scores were different between farms that have a low incidence of PRRS outbreaks, compared to farms that have a high incidence. In the two databases used, the scores consistently showed that farms with higher scores have a higher frequency of PRRS outbreaks. In the first validation, farms that had never had an outbreak investigation before had a significant (p < 0.02) lower score (0.29; 0.21-0.37) when compared to farms that had 2 or more outbreaks (0.43; 0.39-0.46). In the second, the farms of the control group also had significant (p < 0.004) lower scores (0.30; 0.27-0.33) compared to the case group (0.35; 0.33-0.38). Also, the results suggest that events related to swine movements, transmission by air and water, and people movements should be prioritized. The biosecurity vulnerability scores may be useful to assess vulnerabilities on biosecurity protocols in order to reduce the frequency of PRRS outbreaks and may help producers and veterinarians prioritize investments in improving biosecurity practices over time.


Assuntos
Síndrome Respiratória e Reprodutiva Suína/etiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína , Criação de Animais Domésticos/métodos , Animais , Surtos de Doenças/prevenção & controle , Surtos de Doenças/veterinária , Suscetibilidade a Doenças/veterinária , Feminino , Síndrome Respiratória e Reprodutiva Suína/transmissão , Gravidez , Reprodutibilidade dos Testes , Medição de Risco , Fatores de Risco , Inquéritos e Questionários , Suínos
19.
Vet Microbiol ; 223: 27-33, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30173748

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) is the most important disease affecting the swine industry worldwide. Although monocytes and macrophages, especially tissue-resident and alveolar macrophages, are the primary target of PRRSV, monocyte- and bone marrow-derived dendritic cells (DCs) are also susceptible to PRRSV infection. It has been shown that lung DCs cannot be infected with PRRSV, but the response and susceptibility of bona fide conventional DC subtypes (cDCs; cDC1 and cDC2) is unknown. In this work, evaluation of the response of tracheal cDC1 and cDC2 subsets to PRRSV revealed differential cytokine expression, whereby cDC1 subsets expressed higher levels of IFN-α and cDC2 subsets more IL-10. Toll-like receptors (TLRs) were also affected: cDC2 cells induced greater upregulation of TLR2 and TLR4, and CD163+ cells showed TLR3 upregulation. However, we could not demonstrate under our experimental conditions that cDC1 and cCD2 subsets are susceptible to PRRSV infection. Our findings show the effects of PRRSV on cDC1 and cDC2 subsets and that these cells were not infected by PRRSV.


Assuntos
Citocinas/metabolismo , Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/virologia , Macrófagos/imunologia , Macrófagos/virologia , Monócitos/imunologia , Monócitos/virologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Receptores de Superfície Celular/metabolismo , Suínos , Traqueia/imunologia , Traqueia/virologia
20.
Transbound Emerg Dis ; 65(4): 993-1008, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29504285

RESUMO

Porcine reproductive and respiratory syndrome (PRRS) is caused by a genetically diverse RNA virus and is an economically significant disease in the swine industry. In this study, a total of 8,126 serum samples were obtained from 275 technified and semi-technified farms belonging to 30 of the 32 states of Mexico and representative of the eight regions of the country. Anti-PRRSv antibodies against the PRRS vaccine and an isolated wild Mexican virus were tested by ELISA. Antibodies were found in 15%-49% of the tested sera, with 2.4%-9.8% against the vaccine and 7.7%-26% against the wild virus. The PRRSv virus was detected by RT-PCR in 77 of the 1,630 pooled samples tested, representing seven of the eight geographic regions into which the Mexican Republic is divided. The complete sequences of open reading frames 5 and 7 from 20 PRRSv-positive samples were determined. The analysis of the sequences together with the previously published sequences of historic strains revealed that all the strains belonged to the one, five and eight lineages of the PRRSV2. Striking differences, particularly in ORF5 and ORF7, were found between sequences of the strains and the reference virus, due to insertions and substitutions in positions that play key roles in the recognition, structure and function of the virus. Overall, these results established the magnitude of PRRS virus genetic diversity, and the most frequent virus strain that predominates in Mexico. The PRRSV2 is presented in the porcine population of Mexico; the circulating strains have important changes in ORF5 and ORF7, which probably explain the results obtained in the serological analysis of the wild virus and vaccine strains.


Assuntos
Variação Genética , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Proteínas do Envelope Viral/genética , Proteínas Virais/análise , Sequência de Aminoácidos , Criação de Animais Domésticos/métodos , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , México/epidemiologia , Dados de Sequência Molecular , Filogenia , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Estudos Soroepidemiológicos , Suínos , Proteínas Virais/classificação , Proteínas Virais/genética
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