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Chuanqing Wang - One of the best experts on this subject based on the ideXlab platform.

  • enhancing mucosal immunity in mice by recombinant adenovirus expressing major epitopes of Porcine Circovirus 2 capsid protein delivered with cytosine phosphate guanosine oligodeoxynucleotides
    Journal of Veterinary Science, 2014
    Co-Authors: Hongtao Chang, Xiuyuan He, Lu Chen, Qiuying Yu, Jun Zhao, Xinwei Wang, Xia Yang, Chuanqing Wang
    Abstract:

    : A recombinant replication-defective adenovirus expressing the major epitopes of Porcine Circovirus-2 (PCV-2) capsid protein (rAd/Cap/518) was previously constructed and shown to induce mucosal immunity in mice following intranasal delivery. In the present study, immune responses induced by intranasal immunization with a combination of rAd/Cap/518 and cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODN) were evaluated in mice. The levels of PCV-2-specific IgG in serum and IgA in saliva, lung, and intestinal fluids were significantly higher in the group immunized with rAd/Cap/518 and CpG ODN than animals immunized with rAd/Cap/518 alone. The frequencies of IL-2-secreting CD4⁺ T cells and IFN-γ-producing CD8⁺ T cells were significantly higher in the combined immunization group than mice immunized with rAd/Cap/518 alone. The frequencies of CD3⁺, CD3⁺CD4⁺CD8⁻, and CD3⁺CD4⁻CD8⁺ T cells in the combined immunization group were similar to that treated with CpG ODN alone, but significantly higher than mice that did not receive CpG ODN. PCV-2 load after challenge in the combined immunization group was significantly lower than that in the phosphate-buffered saline placebo group and approximately 7-fold lower in the group treated with CpG ODN alone. These results indicate that rAd/Cap/518 combined with CpG ODN can enhance systemic and local mucosal immunity in mice, and represent a promising synergetic mucosal vaccine against PCV-2.

  • induction of mucosal immunity by intranasal immunization with recombinant adenovirus expressing major epitopes of Porcine Circovirus 2 capsid protein
    Veterinary Immunology and Immunopathology, 2013
    Co-Authors: Yufeng Liu, Hongtao Chang, Lu Chen, Jun Zhao, Xinwei Wang, Xia Yang, Quanhai Guo, Chuanqing Wang
    Abstract:

    Porcine Circovirus-2 (PCV-2) is primarily transmitted through mucosa, thus the mucosal immunity may constitute an essential feature of vaccination strategies against PCV-2 infection. Mucosal immunity elicited by recombinant replication-deficient adenovirus expressing the major epitopes of PCV-2 capsid protein (rAd/Cap/518) via intranasal (i.n.), intramuscular (i.m.) or oral routes in mice were evaluated. Immunization with rAd/Cap/518 via i.n. route induced higher titers of IgA in saliva, bronchoalveolar and intestinal lavage fluid compared with those immunized via i.m. route. The proportions of CD3+, CD3+CD4+ and CD3+CD8+ T cells were significantly increased in mice immunized with rAd/Cap/518 via i.n. route compared with the control group. Higher levels of IFN-γ were detected in the spleen and mesenteric lymph nodes of mice immunized with rAd/Cap/518 via i.n. route compared with other groups, yet IL-4 was not detected in any group. Real-time PCR analysis confirmed viral DNA loads in the i.m. or i.n. immunization group was lower than that seen in the rAd immunization. These results indicate that i.n. administration of rAd/Cap/518 can elicit humoral and Th1-type cellular protective immunity in both systemic and mucosal immune compartments in mice, representing a promising mucosal vaccine candidate against PCV-2.

Gonzalo Fernandez - One of the best experts on this subject based on the ideXlab platform.

  • molecular assessment of visitor personal protective equipment contamination with the aleutian mink disease virus and Porcine Circovirus 2 in mink and Porcine farms
    PLOS ONE, 2018
    Co-Authors: Jose Manuel Diaz Cao, Alberto Prieto, Gonzalo Lopez, Ricardo Fernandezantonio, P Diaz, C Lopez, Susana Remesar, P Diezbanos, Gonzalo Fernandez
    Abstract:

    Personal protective equipment (PPE) is an element of biosecurity intended to prevent the access or spread of diseases in farms. Nevertheless, to date no extensive reports exist about the effectiveness of different available PPE on farms. Thus, our aim was to estimate the degree of protection of PPE from viral contamination during farm visits. Two farms, infected with Aleutian mink disease virus and Porcine Circovirus–2 respectively, were visited by six visitors wearing different combinations of PPE: coveralls with hood and bootcovers, both with a certified barrier to infective agents (certified PPE group) and non-certified bootcover and coverall without hood (non-certified PPE group). Seventy-two swab samples from PPE and both hair and street clothes under PPE were taken after the visit and analysed by qPCR. Our results reveal viral exposure during visits, and the external protections of body and shoes were contaminated in all cases (24/24). In addition, protection from viral contamination varied noticeably according to the biosecurity elements used. A higher number of positives were detected in the non-certified PPE group than in the certified PPE group, both in elements under external protections (14/18 vs 3/18) and also in hair (4/6 vs 0/6). In fact, non-certified bootcovers broke during visits, resulting in viral contamination of the internal elements under them; these are consequently not suitable for using with wrinkled surfaces usually found in farm facilities. Thus, certified coveralls should be used in order to prevent contaminations, and workers and personnel of farms should be trained in their proper use. qPCR is a useful tool in the risk management of biosecurity programmes, and our results may serve as a model to evaluate different biosecurity measures.

Joaquim Segales - One of the best experts on this subject based on the ideXlab platform.

  • Genotyping of Porcine Circovirus 2 (PCV-2) in Vaccinated Pigs Suffering from PCV-2-Systemic Disease between 2009 and 2020 in Spain
    'MDPI AG', 2021
    Co-Authors: Marina Sibila, Giovanni Franzo, Mariano Domingo, Caterina Rocco, Eva Huerta, José Ignacio Núñez, Joaquim Segales
    Abstract:

    Vaccination against Porcine Circovirus 2 (PCV-2) is a common practice all over the world. Vaccines can prevent PCV-2-systemic disease (PCV-2-SD) outbreaks but not PCV-2 infection, which can be detectable in a percentage of vaccinated animals. Occasionally, PCV-2-SD is diagnosed in vaccinated farms. The objective of this study was to genotype the PCV-2 strains detected in vaccinated animals diagnosed with PCV-2-SD. Additionally, the evolution of the frequency of PCV-2 genotype detection at Spanish, European, and world levels was assessed. Fifty cases diagnosed as PCV-2-SD between 2009 and 2020 were included in this study. PCV-2 genotype was determined by sequencing the Cap gene region. Among them, only PCV-2b (23/50, 46%) and PCV-2d (27/50, 54%) genotypes were detected. Although the frequency of detection of these two genotypes was similar, their temporal distribution was different. Whereas most PCV-2b sequences (17/23, 74%) were detected between 2009 and 2012, PCV-2d sequences were obtained from 2013 to 2020. Indeed, a predominance of the PCV-2d genotype was observed from 2013 onwards, a trend also noticed at European and world levels. The results suggest that detection of particular genotypes in vaccinated animals probably reflects the general prevalence of the genotypes over time rather than genotype-specific vaccine-immunity escaping

  • Porcine Circovirus 2 pcv 2 genetic variability under natural infection scenario reveals a complex network of viral quasispecies
    Scientific Reports, 2018
    Co-Authors: Florencia Correafiz, Anna Llorens, Giovanni Franzo, Joaquim Segales, Tuija Kekarainen
    Abstract:

    Porcine Circovirus 2 (PCV-2) is a virus characterized by a high evolutionary rate, promoting the potential emergence of different genotypes and strains. Despite the likely relevance in the emergence of new PCV-2 variants, the subtle evolutionary patterns of PCV-2 at the individual-host level or over short transmission chains are still largely unknown. This study aimed to analyze the within-host genetic variability of PCV-2 subpopulations to unravel the forces driving PCV-2 evolution. A longitudinal weekly sampling was conducted on individual animals located in three farms after the first PCV-2 detection. The analysis of polymorphisms evaluated throughout the full PCV-2 genome demonstrated the presence of several single nucleotide polymorphisms (SNPs) especially in the genome region encoding for the capsid gene. The global haplotype reconstruction allowed inferring the virus transmission network over time, suggesting a relevant within-farm circulation. Evidences of co-infection and recombination involving multiple PCV-2 genotypes were found after mixing with pigs originating from other sources. The present study demonstrates the remarkable within-host genetic variability of PCV-2 quasispecies, suggesting the role of the natural selection induced by the host immune response in driving PCV-2 evolution. Moreover, the effect of pig management in multiple genotype coinfections occurrence and recombination likelihood was demonstrated.

  • Genetic variability of Porcine Circovirus 2 in vaccinating and non-vaccinating commercial farms
    Journal of General Virology, 2014
    Co-Authors: Tuija Kekarainen, Anna Llorens, Angel Gonzalez, Joaquim Segales
    Abstract:

    Vaccines against Porcine Circovirus 2 (PCV2) are now widely used to control the diseases caused by the virus. Although the vaccines protect pigs against the disease, they do not lead to sterilizing immunity and therefore infections with PCV2 continue in farms. It is expected that, due to its high evolutionary rate, PCV2 can adapt quickly to environmental pressures such as vaccination. The goal of this study was to elucidate the molecular variation of PCV2 in relation to vaccination. PCV2 variability was investigated from samples of infected pigs from five farms where vaccination had never been applied and two farms where pigs had been vaccinated for at least 2 years. For the genetic analysis, full PCV2 genomes were amplified and subsequently pooled by vaccination status from serum of eight vaccinated, infected pigs and 16 non-vaccinated, infected pigs. Variability of viral populations was quantified using next-generation sequencing and subsequent bioinformatics analysis. The number of segregating sites was similar in the non-vaccinated (n = 109) and vaccinated pools (n = 96), but the distribution of these sites in the genome differed. Most notably, in the capsid gene, the number of segregating sites was observed only in the non-vaccinated population. Based on the structural analysis, it is expected that some low-frequency amino acids result in biologically low-fit viruses. On the contrary, D294 in replicase represents a novel amino acid which was dominant and unique in the vaccinated pool. This work showed that variable PCV2 populations were circulating in commercial farms, and that this variability was different in samples obtained from vaccinating and non-vaccinating farms.

  • experimental inoculation of conventional pigs with Porcine reproductive and respiratory syndrome virus and Porcine Circovirus 2
    Journal of Virology, 2002
    Co-Authors: Albe Rovira, Joaquim Segales, Monica Alasch, L Garcia, J Planadura, C Rosell, H Ellerbrok, Annette Mankertz, Mariano Domingo
    Abstract:

    Postweaning multisystemic wasting syndrome (PMWS) is a disease of nursery and fattening pigs characterized by growth retardation, paleness of the skin, dyspnea, and increased mortality rates. Porcine Circovirus 2 (PCV2) has been demonstrated to be the cause of PMWS. However, other factors are needed for full development of the syndrome, and Porcine reproductive and respiratory syndrome virus (PRRSV) infection has been suggested to be one of them. Twenty-four conventional 5-week-old pigs were distributed in four groups: control (n = 5), PRRSV inoculated (n = 5), PCV2 inoculated (n = 7), and PRRSV and PCV2 inoculated (n = 7). The two groups inoculated with PRRSV showed growth retardation. Pigs inoculated with both PRRSV and PCV2 had increased rectal temperature. One of these pigs developed wasting, had severe respiratory distress, and died. The most important microscopic lesion in pigs inoculated with PCV2 was lymphocyte depletion with histiocytic infiltration of the lymphoid organs, more severe and in a wider range of tissues in doubly inoculated pigs. Interstitial pneumonia was observed in the three inoculated groups. PCV2 nucleic acid was found by in situ hybridization in larger amounts and in a wider range of lymphoid tissues in PRRSV- and PCV2-inoculated than in PCV2-inoculated pigs. TaqMan PCR was performed to quantify the PCV2 loads in serum during the experiment. PCV2 loads were higher in doubly inoculated pigs than in pigs inoculated with PCV2 alone. These findings indicate that severe disease can be reproduced in conventional 5-week-old pigs by inoculation of PRRSV and PCV2. Moreover, these results support the hypothesis that PRRSV infection enhances PCV2 replication.

Jose Manuel Diaz Cao - One of the best experts on this subject based on the ideXlab platform.

  • molecular assessment of visitor personal protective equipment contamination with the aleutian mink disease virus and Porcine Circovirus 2 in mink and Porcine farms
    PLOS ONE, 2018
    Co-Authors: Jose Manuel Diaz Cao, Alberto Prieto, Gonzalo Lopez, Ricardo Fernandezantonio, P Diaz, C Lopez, Susana Remesar, P Diezbanos, Gonzalo Fernandez
    Abstract:

    Personal protective equipment (PPE) is an element of biosecurity intended to prevent the access or spread of diseases in farms. Nevertheless, to date no extensive reports exist about the effectiveness of different available PPE on farms. Thus, our aim was to estimate the degree of protection of PPE from viral contamination during farm visits. Two farms, infected with Aleutian mink disease virus and Porcine Circovirus–2 respectively, were visited by six visitors wearing different combinations of PPE: coveralls with hood and bootcovers, both with a certified barrier to infective agents (certified PPE group) and non-certified bootcover and coverall without hood (non-certified PPE group). Seventy-two swab samples from PPE and both hair and street clothes under PPE were taken after the visit and analysed by qPCR. Our results reveal viral exposure during visits, and the external protections of body and shoes were contaminated in all cases (24/24). In addition, protection from viral contamination varied noticeably according to the biosecurity elements used. A higher number of positives were detected in the non-certified PPE group than in the certified PPE group, both in elements under external protections (14/18 vs 3/18) and also in hair (4/6 vs 0/6). In fact, non-certified bootcovers broke during visits, resulting in viral contamination of the internal elements under them; these are consequently not suitable for using with wrinkled surfaces usually found in farm facilities. Thus, certified coveralls should be used in order to prevent contaminations, and workers and personnel of farms should be trained in their proper use. qPCR is a useful tool in the risk management of biosecurity programmes, and our results may serve as a model to evaluate different biosecurity measures.

C Chae - One of the best experts on this subject based on the ideXlab platform.

  • comparative efficacy of commercial mycoplasma hyopneumoniae and Porcine Circovirus 2 pcv2 vaccines in pigs experimentally infected with m hyopneumoniae and pcv2
    Vaccine, 2011
    Co-Authors: Dong Hyun Kim, C H Kim, Kiwon Han, Hwi Won Seo, Changhoon Park, I Kang, C Chae
    Abstract:

    The efficacies of two commercial Mycoplasma hyopneumoniae bacterins and Porcine Circovirus 2 (PCV2) vaccines were compared in conventional pigs immunized at different ages based on humoral response, pathological observation, and growth performance from birth to finishing (175 days of age) using a M. hyopneumoniae and PCV2 co-infection challenge model. One-week-old pigs (n=110) were randomly assigned to five groups: three vaccinated and challenged (VC), and one each of non-vaccinated and challenged (NVC) and negative control. A significant difference was found in the number of genomic copies of M. hyopneumoniae in nasal swabs and PCV2 in serum samples, the average daily weight gain (gram/pig/day) between 63 and 133 dpi, gross and histopathological lung lesion scores, histopathological lymph node lesion scores, and the immunohistochemical analysis of PCV2 among the three VC groups. The single dose schedule for M. hyopneumoniae bacterins and PCV2 vaccines have the advantages of (i) improving daily weight gain (122.4%) and slaughter weight (120.5%), and (ii) reducing the incidence of clinical signs and lung and lymph node lesions.

  • reproduction of postweaning multisystemic wasting syndrome in pigs by prenatal Porcine Circovirus 2 infection and postnatal Porcine parvovirus infection or immunostimulation
    Veterinary Pathology, 2008
    Co-Authors: Y H Lee, K K Ahn, Bongtae Kim, C Chae
    Abstract:

    Postweaning multisystemic wasting syndrome (PMWS) was reproduced in prenatally Porcine Circovirus 2 (PCV2)-infected pigs by either postnatal infection with Porcine parvovirus (PPV) or by immunostimulation. Twenty-four randomly selected piglets from 3 sows, which had been experimentally infected during gestation with PCV2, were randomly divided into 3 groups; group 1 (prenatal PCV2 infection, with postnatal PPV infection), group 2 (prenatal PCV2 infection, with postnatal keyhole limpet hemocyanin, emulsified in incomplete Freund's adjuvant [KLH/ICFA] injection), and group 3 (prenatal PCV2 infection only). Twenty-four randomly selected piglets from 3 uninfected sows were randomly divided into 3 groups; group 4 (no prenatal infection, with postnatal PCV2 and PPV infection), group 5 (no prenatal infection, with postnatal PCV2 infection), and group 6 (negative control pigs). Body weight in negative control pigs (group 6) was increased significantly compared with pigs in groups 1, 2, and 4 at 49, 52, 56, 59, and 63 days of age. The granulomatous inflammatory reaction and lymphoid depletion that are typical lesions in pigs with PMWS were observed in the lymph node of piglets in groups 1, 2, and 4 at 63 days of age. Pigs in group 3 had significantly fewer PCV2-positive cells than those from groups 1, 2, 4, or 5. When the prenatally PCV2-infected pigs were infected with PPV or injected with immunostimulant in the postnatal period, they developed PMWS. Thus, factors that potentiate the progression of prenatal PCV2 infection to PMWS are postnatal infection with PPV or immune stimulation.

  • expression of monocyte chemoattractant protein 1 and macrophage inflammatory protein 1 in Porcine Circovirus 2 induced granulomatous inflammation
    Journal of Comparative Pathology, 2004
    Co-Authors: Ji Man Kim, C Chae
    Abstract:

    The expression of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1 (MIP-1) in granulomatous inflammation of lymph nodes was determined in 1-day-old piglets inoculated intranasally with Porcine Circovirus 2 (PCV2). The methods used were reverse transcription-polymerase chain reaction (RT-PCR), granulomatous lesion scoring, and in-situ hybridization. Maximum MCP-1 expression and MIP-1 expression were observed at 17 and 21 days post-inoculation (dpi), respectively. Granulomatous lesions of maximum severity were observed somewhat later (28 dpi). A close cell to cell relation between PCV2 and the two cytokines in serial sections from lymph nodes suggested that their induction was a direct response to PCV2. The results provide evidence of the ability of MCP-1 and MIP-1 to induce granulomatous inflammation in vivo.

  • pathogenesis of postweaning multisystemic wasting syndrome reproduced by co infection with korean isolates of Porcine Circovirus 2 and Porcine parvovirus
    Journal of Comparative Pathology, 2003
    Co-Authors: Ji Man Kim, C Choi, C Chae
    Abstract:

    Thirty-two colostrum-deprived conventional pigs aged 28 days were co-infected with Korean isolates of Porcine Circovirus 2 (PCV2) and Porcine parvovirus (PPV) and observed for up to 35 days post-inoculation (dpi). Histopathologically, granulomatous inflammation, with or without intracytoplasmic inclusion bodies, was present in lymphoid tissues (e.g., lymph nodes, thymus, spleen and tonsil) from 20 dpi, being most severe at 24 dpi. PCV2 and PPV DNA were detected in the lymph nodes from 3 to 35 dpi by in-situ hybridization, but the labelling for both viruses was particularly intense and widespread at 20 and 24 dpi. A close relationship between the cells labelled for PCV2 and those labelled for PPV was revealed by examination of serial sections from lymph nodes, and PCV2 and PPV were also detected in peripheral blood monocytes from 3 to 35 dpi. Other tissues and cells in which PCV2 and PPV DNA were detected included macrophages in the tonsil, thymus, spleen, lung, liver, kidney and heart. Significantly more PCV2-positive cells than PPV-positive cells were detected in the lymph nodes at 5 to 35 dpi. The pathogenesis of postweaning multisystemic wasting syndrome (PMWS) reproduced in this study may be suggested thus: initial viral entry through tonsillar macrophages, followed within 3 days by viraemia; PCV2 and PPV replicate, at least to some extent, in circulating peripheral monocytes, contributing to the cell-associated viraemia and to viral distribution throughout the lymphoid tissues.