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Point-of-service diagnostic technology for detection of swine viral diseases
2020
Nannucci, Lapo | Barattini, Paolo | Bossis, Ioannis | Woźniakowski, Grzegorz | Balka, Gyula | Pugliese, Carolina
A research project is underway aiming to develop a field diagnostic tool for six important viruses of the pig sector, namely: African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), porcine parvovirus (PPV), porcine circovirus (PCV2), and classical swine fever virus (CSFV). To obtain a preliminary sounding of the interest in this type of instrument among its potential operators, a questionnaire was drawn up and submitted to three categories of stakeholders: farmers, veterinarians, and others (including scientific and technical staff working on animal farms). Four countries participated: Italy, Greece, Hungary, and Poland. In total, 83 replies were collected and analysed in a breakdown by stakeholder type and pertinence, where the areas were the importance of the main diseases within the different countries, diagnostic tool operational issues, and economic issues. The main end-users of this kind of instrument are expected to be private veterinarians and pig producers. The infectious agents seeming to be most interesting to diagnose with the instrument are PRRSV, SIV, PPV, and PCV2. The most decisive parameters which have been selected by the stakeholders are sensitivity, cost, simplicity, and time required to obtain results. The economic issue analysis showed that the majority of those who would prefer to buy rather than rent the device are willing to pay up to €3,000 for a diagnostic field tool.
Mostrar más [+] Menos [-]Cloning and identification of PK15 cells for enhanced replication of classical swine fever virus
2020
Yin Mei | Hu Dongfang | Li Peng | Kong Lingyun | Ning Hongmei | Yue Feng | Jiang Jinqing | Wang Xuannian
Classical swine fever virus (CSFV) causes an economically important and highly contagious disease of pigs, leading to economic losses around the world. Attenuated live vaccines with CSFV antigens have played an important role in the prevention and control of the disease. Porcine kidney 15 (PK15) cells have been widely used for the propagation of CSFV, but this cell line is not efficient or homogeneously susceptible to viral infection.
Mostrar más [+] Menos [-]Characterisation of newly emerged isolates of classical swine fever virus in China, 2014–2015
2017
Leng, Chaoliang | Zhang, Hongliang | Kan, Yunchao | Yao, Lunguang | Li, Mingliang | Zhai, Hongyue | Li, Zhen | Liu, Chunxiao | Shi, Hongfei | Ji, Jun | Qiu, Reng | Tian, Zhijun
Introduction: In 2014–2015, the epidemic of classical swine fever (CSF) occurred in many large-scale pig farms in different provinces of China, and a subgenotype 2.1d of CSF virus (CSFV) was newly identified. Material and Methods: The phylogenetic relationship, genetic diversity, and epidemic status of the 2014–2015 CSFV isolates, 18 new CSFV isolates collected in 2015, and 43 other strains isolated in 2014–2015 were fully analysed, together with 163 CSFV reference isolates. Results: Fifty-two 2014–2015 isolates belonged to subgenotype 2.1d and nine other isolates belonged to subgenotype 2.1b. The two subgenotype isolates showed unique molecular characteristics. Furthermore, the 2.1d isolates were found to possibly diverge from 2.1b isolates. Conclusion: This study suggests that the Chinese CSFVs will remain pandemic.
Mostrar más [+] Menos [-]Cloning and identification of PK15 cells for enhanced replication of classical swine fever virus
2020
Yin, Mei | Hu, Dongfang | Li, Peng | Kong, Lingyun | Ning, Hongmei | Yue, Feng | Jiang, Jinqing | Wang, Xuannian
Classical swine fever virus (CSFV) causes an economically important and highly contagious disease of pigs, leading to economic losses around the world. Attenuated live vaccines with CSFV antigens have played an important role in the prevention and control of the disease. Porcine kidney 15 (PK15) cells have been widely used for the propagation of CSFV, but this cell line is not efficient or homogeneously susceptible to viral infection. To achieve a homogeneous PK15 cell line which enabled high titre replication of CSFV, we used the limiting dilution cell cloning method. We developed two cell clones, PK15-1A6 and PK15-3B1, which respectively have high- and low-permissive phenotypes to CSFV infection. The PK15-1A6, PK15-3B1, and PK15 parent cells showed different characteristics in cell proliferation rate, susceptibility to CSFV infection, and CSFV production. The mean virus titres per millilitre reflected by TCID₅₀ values in PK15-1A6, PK15-3B1, and PK15 parent cells were 106.85, 103.63, and 104.74, respectively. The PK15-1A6 cell clone is more permissive to CSFV infection than the PK15 parent cells. The screened high-permissive cells will be useful for CSFV propagation and vaccine development in vitro, and facilitate research on the pathogenicity of CSFV.
Mostrar más [+] Menos [-]Complete genome sequences of three sub-genotype 2.1b isolates of classical swine fever virus in China
2018
Liu Chunxiao | Li Mingliang | Yin Xingwang | Zhang Hongliang | Xiang Lirun | Zhai Hongyue | Wang Congcong | Kan Yunchao | Yao Lunguang | Tian Zhijun | Leng Chaoliang
Classical swine fever (CSF) has caused severe economic losses in pig production in many countries. Recent CSF outbreaks in China are mainly associated with sub-genotype 2.1 of CSF virus (CSFV). Although there is abundant information regarding 2.1 isolates, few data are available on whole-genome analysis.
Mostrar más [+] Menos [-]Characterisation of newly emerged isolates of classical swine fever virus in China, 2014–2015
2017
Leng Chaoliang | Zhang Hongliang | Kan Yunchao | Yao Lunguang | Li Mingliang | Zhai Hongyue | Li Zhen | Liu Chunxiao | Shi Hongfei | Ji Jun | Qiu Reng | Tian Zhijun
Introduction: In 2014–2015, the epidemic of classical swine fever (CSF) occurred in many large-scale pig farms in different provinces of China, and a subgenotype 2.1d of CSF virus (CSFV) was newly identified.
Mostrar más [+] Menos [-]Establishment of a method for evaluation of the efficacy of a classical swine fever virus subunit vaccine in rabbits
2020
Cao, Zhi | Zhang, Heng | Yang, Qian | Zhang, Hui | Fan, Gencheng
OBJECTIVE To establish a method for evaluation of the efficacy of a classical swine fever virus (CSFV) subunit vaccine in rabbits as determined via humoral immune responses to the virus. ANIMALS 40 specific pathogen–free rabbits. PROCEDURES Rabbits were randomly assigned to 4 groups (10 rabbits/group) for SC injection of 0.05, 0.1, and 0.2 mL of a CSFV subunit E2 vaccine (representing 1.15, 2.3, or 4.6 μg of E2 protein/dose, respectively) or saline (0.9% NaCl) solution. Blood samples were collected 21 days after vaccination for measurement of the antibody response against CSFV via ELISA and virus neutralization methods. On the same day, the CSFV Chinese (C) strain was injected into an ear vein. Vaccine efficacy was determined by monitoring of rabbits for pyrexia for 4 days and measurement of viral copies in spleen lysates at the end of the study. Reproducibility of the antibody response was tested with 2 other batches of the vaccine at the minimum immunization dose identified for the initially tested batch. RESULTS The E2 protein dose of the initially tested vaccine was positively correlated with the antibody response and protection rate in rabbits. The identified minimum immunization dose per rabbit was 0.1 mL, representing an E2 protein content of approximately 2.3 μg, and reproducibility of the antibody response to vaccination with the 2 other batches at this dose was good. CONCLUSIONS AND CLINICAL RELEVANCE A method was established in rabbits for evaluation of the efficacy of a CSFV subunit vaccine that could help in the optimization of later large-scale vaccine production and quality control processes as well as in the clinical application of the vaccine.
Mostrar más [+] Menos [-]Suppression of immune responses in pigs by nonstructural protein 1 of Porcine reproductive and respiratory syndrome virus
2012
Zhou, Yefei | Bai, Juan | Li, Yufeng | Wang, Xinglong | Wang, Xianwei | Jiang, Ping
Porcine reproductive and respiratory syndrome (PRRS) is characterized by a delayed and defective adaptive immune response. The viral nonstructural protein 1 (NSP1) of the PRRS virus (PRRSV) is able to suppress the type I interferon (IFN) response in vitro. In this study, recombinant adenoviruses (rAds) expressing NSP1 (rAd-NSP1), glycoprotein 5 (GP5) (rAd-GP5), and the NSP1-GP5 fusion protein (rAd-NSP1-GP5) were constructed, and the effect of NSP1 on immune responses was investigated in pigs. Pigs inoculated with rAd-NSP1 or rAd-NSP1-GP5 had significantly lower levels of IFN-γ and higher levels of the immunosuppressive cytokine IL-10 than pigs inoculated with rAd-GP5, wild-type adenovirus, or cell culture medium alone. The antibody response to vaccination against classic swine fever virus (CSFV) was significantly decreased by inoculation of NSP1 7 d after CSFV vaccination in pigs. Thus, NSP1-mediated immune suppression may play an important role in PRRSV pathogenesis.
Mostrar más [+] Menos [-]Susceptibility of piglets to rabbit hemorrhagic disease virus following experimental infection
2000
Shien, J. H. | Lee, L. H.
The possibility exists that rabbit hemorrhagic disease virus (RHDV) can be transmitted to swine, through lapinized hog cholera virus (HCV) vaccine. To investigate the infectivity of RHDV in swine, 16 four- to six-week-old piglets were inoculated subcutaneously with RHDV, and samples of liver, lung, spleen, kidney, bile, adrenal gland, tonsil, mesenteric lymph node, thymus, urine, buffy coat, and feces were collected from each of 2 animals on Days 0, 1, 2, 3, 5, 7, 14, and 28 post infection. Using reverse transcription-polymerase chain reaction, viral RNA was detected in most tissues by Day 3 and was absent after Day 5, except in lung and liver tissues, in which viral RNA was detected up to Day 14. Viral RNA was not detected in kidney, urine, feces or bile. Antibody responses, as detected by hemagglutination inhibition, were of low titer and short duration, and were similar in animals inoculated with viable RHD and in those given formalin-inactivated RHDV (n = 2). Neither viral RNA nor antibody were detected in the negative control or in the uninfected, in-contact animals.
Mostrar más [+] Menos [-]Analysis of in vitro apoptosis induced by virulent Korean isolate of classical swine fever virus in peripheral blood B cell line
2012
Kim, S.M., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Lim, S.I., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Song, J.Y., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Hyun, B.H., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea
Classical swine fever (CSF) is a highly contagious disease among swine that has an important economic impact on worldwide. One clinical symptom of CSF is leukopenia, in particular lymphopenia, which is a characteristic event that occurs early in the course of CSF. Though lymphopenia associated with apoptosis, the pathogenic mechanism underlying the lymphopenia has not been well studied. To understand these mechanisms, we investigated the response of porcine B cell lines to infection with SW03, virulent strain isolated from swine tissue in Korea. This study demonstrated that SW03-infected L35 cell were induced apoptosis through the detection of activated caspase-3. In addition, SW03 infection leaded to alterations in pro-apoptotic, Bax, and anti-apoptotic, Bcl-xL proteins of Bcl-2 family. Our results would suggest that SW03-infected L35 cells induced apoptosis via intrinsic mitochondrial pathway.
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