pathological changes in masked palm civets experimentally infected by severe acute CORD-Papers (Version 1)

Title: Pathological Changes in Masked Palm Civets Experimentally Infected by Severe Acute Respiratory Syndrome (SARS) Coronavirus
Abstract: Masked palm civets are highly susceptible to infection with the severe acute respiratory syndrome coronavirus (SARS-CoV). Infected animals become less aggressive and develop pyrexia lethargy and diarrhoea. The present study describes the spectrum of histopathological changes in the lung spleen lymph node liver small intestine kidney and cerebrum of civets infected experimentally with SARS-CoV. In-situ hybridization (ISH) with probes specific for the RNA polymerase gene demonstrated viral RNA in the lung small intestine and cerebrum only. In-situ labelling was employed in order to demonstrate cellular apoptosis in the cerebrum but there was no evidence of apoptosis within the myocardium. These results indicate that SARS-CoV causes multi-organ pathology in civets similar to that observed in human SARS patients. These parallels suggest that civets may be used as an animal model of this infection to gain insight into the pathogenesis of SARS and for evaluation of candidate vaccines and antiviral drugs.
Published: 2008-03-17
Journal: J Comp Pathol
DOI: 10.1016/j.jcpa.2007.12.005
DOI_URL: http://doi.org/10.1016/j.jcpa.2007.12.005
Author Name: Xiao Y
Author link: https://covid19-data.nist.gov/pid/rest/local/author/xiao_y
Author Name: Meng Q
Author link: https://covid19-data.nist.gov/pid/rest/local/author/meng_q
Author Name: Yin X
Author link: https://covid19-data.nist.gov/pid/rest/local/author/yin_x
Author Name: Guan Y
Author link: https://covid19-data.nist.gov/pid/rest/local/author/guan_y
Author Name: Liu Y
Author link: https://covid19-data.nist.gov/pid/rest/local/author/liu_y
Author Name: Li C
Author link: https://covid19-data.nist.gov/pid/rest/local/author/li_c
Author Name: Wang M
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wang_m
Author Name: Liu G
Author link: https://covid19-data.nist.gov/pid/rest/local/author/liu_g
Author Name: Tong T
Author link: https://covid19-data.nist.gov/pid/rest/local/author/tong_t
Author Name: Wang L F
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wang_l_f
Author Name: Kong X
Author link: https://covid19-data.nist.gov/pid/rest/local/author/kong_x
Author Name: Wu D
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wu_d
sha: ed9f973784cad4c42001deddab29e9e68d7e4283
license: no-cc
license_url: [no creative commons license associated]
source_x: Elsevier; Medline; PMC
source_x_url: https://www.elsevier.com/https://www.medline.com/https://www.ncbi.nlm.nih.gov/pubmed/
pubmed_id: 18343398
pubmed_id_url: https://www.ncbi.nlm.nih.gov/pubmed/18343398
pmcid: PMC7094611
pmcid_url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7094611
url: https://www.ncbi.nlm.nih.gov/pubmed/18343398/ https://doi.org/10.1016/j.jcpa.2007.12.005 https://api.elsevier.com/content/article/pii/S0021997508000029 https://www.sciencedirect.com/science/article/pii/S0021997508000029
has_full_text: TRUE
Keywords Extracted from Text Content: liver human SARS patients cellular lymph node kidney cerebrum spleen SARS-CoV lung multi-organ pathology myocardium coronavirus SARS-CoV. civets nerve cells human SARS erythrocytes neurons N-lauroylsarcosine SARS-CoV isolates lymph nodes myocardium lamina propria bronchiolar epithelial cells terminal deoxynucleotidyl transferase splenic farms civets Days cynomolgus monkeys pneumocytes lining Tissue human patients pulmonary cerebrum Lymph Nodes Kidneys pancreas Cerebrum Â0.2 SARS-CoV genome sequences lymphoid follicles liver ferrets anti-digoxigenin antibody alveolar spaces cytoplasm monkey tissues stomach foci lung trachea saline coronavirus fur SARS-CoV. DAB non-human primates ethanol tissue sections hamsters SARS coronavirus human heart cellular lymph node Fig. 1c ACE2 receptor molecule human SARS patients intestine cardiac tissues lumina veins kidney chickens splenic tissue hepatic lobules renal cortex sodium dodecyl sulphate mice organs individuals patients cell follicular marmosets avidin-peroxidase conjugate small intestine pulp lymphoid aggregates neuronal peroxidase SARS-CoV lesions Guangzhou Spleen Harbin Veterinary Research Institute, glands bronchioles Liver Â0.5 alveolar portal albino hamsters green monkeys spleen H 2 O 2 rats California Wang SARS-CoV. Bats humans human ACE2 carnivores alveolar epithelia tissues Vero E6 cells Vero E6 cell lungs proteinase K. civets BJ01 pulmonary interstitium purchased blood cell saline citrate sperm DNA alveoli glial cells human SARS-CoV tissue cerebral Serology macaques GZ01 macrophages rhesus macaques monkeys Mayer's haematoxylin alveolar septae mouse sections B guinea-pigs palm civets vessels pulp cats diaminobenzidine tetrahydrochloride bats cells lymphocytes BJ01 Vero E6 cell line SARS-CoV-like virus GZ01 SARS-CoV isolates
Extracted Text Content in Record: First 5000 Characters:Masked palm civets are highly susceptible to infection with the severe acute respiratory syndrome coronavirus (SARS-CoV). Infected animals become less aggressive and develop pyrexia, lethargy and diarrhoea. The present study describes the spectrum of histopathological changes in the lung, spleen, lymph node, liver, small intestine, kidney and cerebrum of civets infected experimentally with SARS-CoV. In-situ hybridization (ISH) with probes specific for the RNA polymerase gene demonstrated viral RNA in the lung, small intestine and cerebrum only. In-situ labelling was employed in order to demonstrate cellular apoptosis in the cerebrum, but there was no evidence of apoptosis within the myocardium. These results indicate that SARS-CoV causes multi-organ pathology in civets, similar to that observed in human SARS patients. These parallels suggest that civets may be used as an animal model of this infection to gain insight into the pathogenesis of SARS and for evaluation of candidate vaccines and antiviral drugs. The aetiological agent of this syndrome was a newly emerged and previously unrecognized coronavirus, now known as SARS coronavirus (SARS-CoV) Ksiazek et al., 2003) . SARS is an acute pulmonary syndrome characterized by atypical pneumonia, progressive respiratory failure and death in up to 10% of infected individuals (Poon et al., 2004) . Although the SARS epidemic has subsided, many authorities, including the World Health Organization (WHO) and the US Centers for Disease Control and Prevention (CDC), have warned of the possible re-emergence of this highly infectious disease. It is therefore imperative that effective measures to prevent and treat the disease are developed and evaluated. To achieve this goal, animal models will play an essential role in studying the pathogenesis of SARS-CoV infection and in developing effective vaccines and therapeutics. A wide range of animal species has been confirmed to be susceptible to experimental infection with SARS-CoV, including rodents (mice and hamsters), carnivores (ferrets and cats) and non-human primates (Wang et al., 2006) . Adult mice infected with SARS-CoV via the respiratory tract show no clinical signs of disease and only mild respiratory tract inflammation . Aged mice, hamsters and ferrets do show signs of clinical disease such as weight loss and ruffled fur but do not develop lung pathology (Martina et al., 2003; Roberts et al., 2005a,b) . Two groups of investigators have studied SARS-CoV infection in African green monkeys and common marmosets McAuliffe et al., 2004; Greenough et al., 2005) . Others have evaluated cynomolgus monkeys and rhesus macaques as potential models, but clinical disease is inconsistently induced in these latter species McAuliffe et al., 2004; Rowe et al., 2004) . Our laboratory has evaluated the suitability of guinea-pigs, hamsters, albino hamsters, chickens and rats as experimental models following inoculation of SARS-CoV strain BJ01. No clinical signs or tissue histopathological changes were observed in any of these animals post-infection. By contrast, all cynomolgus monkeys and rhesus macaques inoculated in this manner developed interstitial pneumonia of variable severity but no other tissue changes . This pulmonary pathology was similar in nature to that seen in SARS patients, but the lesions were less severe than in infected humans. Therefore, there remains a requirement for an animal model of human SARS that closer approximates the tissue pathology that occurs in the human disease. In October 2003 it was reported that SARS-CoVlike viruses had been isolated from masked palm civets from a live animal market in Guangdong, China (Guan et al., 2003) . This report prompted us to examine the feasibility of using civets as a better animal model for SARS. In a preliminary infection study, two groups of civets were inoculated with two different strains of SARS-CoV. Strain GZ01 was isolated during the early stages of the SARS epidemic and strain BJ01 was isolated during the middle phase of the epidemic (Wu et al., 2005) . Both strains were shown capable of infecting civets and inducing clinical signs including pyrexia, lethargy and loss of aggression (Wu et al., 2005) . The aim of the present study was to extend these observations by characterizing the tissue pathology in the infected civets and determining the distribution of viral RNA in tissues from those animals. In addition, analysis of cellular apoptosis in selected tissues was also conducted in an attempt to understand the pathogenesis of this viral infection in civets and to further determine the suitability of this species to model the human disease. The animals and groups used in this study correspond exactly to those described in a previous publication (Wu et al., 2005) . SARS-CoV isolates were propagated in Vero E6 cells for two additional passages to generate virus stocks with titres of 1 Â 10 6 50% tissue culture infective doses (TCID 50 )/ml. Ten one-yearold masked pal
Keywords Extracted from PMC Text: CTG ATT TAG avidin-peroxidase conjugate portal alveolar epithelia alveolar spaces farms cynomolgus monkeys sodium dodecyl sulphate albino hamsters hepatic lobules monkeys interstitial inflammatory infiltrates human ACE2 California cytoplasm human SARS-CoV SARS-CoV isolates SARS-CoV. oedema bronchiolar epithelial cells cell Fig. 1c glial cells macrophages erythrocytes mice patients veins Harbin Veterinary Research Institute, BJ01 cerebral lamina propria lungs liver human alveolar septae small intestine rhesus macaques organs follicular ACE2 receptor molecule Guangzhou pneumocytes lining human patients sections × Fouchier GTC-3′ Vero E6 cell bronchioles civets carnivores ctg cta aag cat ata agg att acc tag-3′ Mayer's haematoxylin nerve cells alveolar Probe 2 ethanol GZ01 diaminobenzidine tetrahydrochloride non-human primates ×0.2 People's Republic pancreas ×0.5 lymph nodes anti-digoxigenin antibody SARS coronavirus (TCID50)/ml spleen neuronal cats blood cell tissue intestine SARS-CoV genome sequences tissues SARS-CoV. Bats saline individuals vessels palm civets rats macaques cardiac tissues Vero E6 cells SARS-CoV-like viruses hamsters DAB splenic tissue human SARS patients stomach marmosets fur renal cortex bats heart lung Wang N-lauroylsarcosine glands saline citrate pulmonary ferrets myocardium chickens lymph node lymphocytes pulmonary interstitium lymphoid follicles peroxidase green monkeys cellular proteinase K. mouse trachea purchased sperm DNA pulp lymphoid aggregates Serology ×2 SARS-CoV splenic lumina humans cells monkey tissues B foci coronavirus neurons guinea-pigs lesions tissue sections Cerebrum alveoli terminal deoxynucleotidyl transferase human SARS pulp cerebrum kidney
Extracted PMC Text Content in Record: First 5000 Characters:Severe acute respiratory syndrome (SARS) first emerged in Guangdong Province in the People's Republic of China in November 2002. The aetiological agent of this syndrome was a newly emerged and previously unrecognized coronavirus, now known as SARS coronavirus (SARS-CoV) (Kuiken et al., 2003, Ksiazek et al., 2003). SARS is an acute pulmonary syndrome characterized by atypical pneumonia, progressive respiratory failure and death in up to 10% of infected individuals (Poon et al., 2004). Although the SARS epidemic has subsided, many authorities, including the World Health Organization (WHO) and the US Centers for Disease Control and Prevention (CDC), have warned of the possible re-emergence of this highly infectious disease. It is therefore imperative that effective measures to prevent and treat the disease are developed and evaluated. To achieve this goal, animal models will play an essential role in studying the pathogenesis of SARS-CoV infection and in developing effective vaccines and therapeutics. A wide range of animal species has been confirmed to be susceptible to experimental infection with SARS-CoV, including rodents (mice and hamsters), carnivores (ferrets and cats) and non-human primates (Wang et al., 2006). Adult mice infected with SARS-CoV via the respiratory tract show no clinical signs of disease and only mild respiratory tract inflammation (Subbarao et al., 2004). Aged mice, hamsters and ferrets do show signs of clinical disease such as weight loss and ruffled fur but do not develop lung pathology (Martina et al., 2003, Roberts et al., 2005a, Roberts et al., 2005b). Two groups of investigators have studied SARS-CoV infection in African green monkeys and common marmosets (Kuiken et al., 2003, McAuliffe et al., 2004, Greenough et al., 2005). Others have evaluated cynomolgus monkeys and rhesus macaques as potential models, but clinical disease is inconsistently induced in these latter species (Kuiken et al., 2003, McAuliffe et al., 2004, Rowe et al., 2004). Our laboratory has evaluated the suitability of guinea-pigs, hamsters, albino hamsters, chickens and rats as experimental models following inoculation of SARS-CoV strain BJ01. No clinical signs or tissue histopathological changes were observed in any of these animals post-infection. By contrast, all cynomolgus monkeys and rhesus macaques inoculated in this manner developed interstitial pneumonia of variable severity but no other tissue changes (Liu et al., 2004). This pulmonary pathology was similar in nature to that seen in SARS patients, but the lesions were less severe than in infected humans. Therefore, there remains a requirement for an animal model of human SARS that closer approximates the tissue pathology that occurs in the human disease. In October 2003 it was reported that SARS-CoV-like viruses had been isolated from masked palm civets from a live animal market in Guangdong, China (Guan et al., 2003). This report prompted us to examine the feasibility of using civets as a better animal model for SARS. In a preliminary infection study, two groups of civets were inoculated with two different strains of SARS-CoV. Strain GZ01 was isolated during the early stages of the SARS epidemic and strain BJ01 was isolated during the middle phase of the epidemic (Wu et al., 2005). Both strains were shown capable of infecting civets and inducing clinical signs including pyrexia, lethargy and loss of aggression (Wu et al., 2005). The aim of the present study was to extend these observations by characterizing the tissue pathology in the infected civets and determining the distribution of viral RNA in tissues from those animals. In addition, analysis of cellular apoptosis in selected tissues was also conducted in an attempt to understand the pathogenesis of this viral infection in civets and to further determine the suitability of this species to model the human disease. The animals and groups used in this study correspond exactly to those described in a previous publication (Wu et al., 2005). SARS-CoV isolates were propagated in Vero E6 cells for two additional passages to generate virus stocks with titres of 1 × 106 50% tissue culture infective doses (TCID50)/ml. Ten one-year-old masked palm civets were housed in individual biosafety isolators and were divided into two groups (n = 5 per group). Animals in groups A and B were inoculated with 3 ml of virus solution containing 3 × 106 TCID50 of BJ01 and GZ01 isolates, respectively, with 2 ml instilled into the trachea and 1 ml given intranasally. A control civet was mock-infected in an identical fashion with 3 ml of Vero E6 cell culture supernatant. All work with infectious virus was performed inside a biosafety cabinet, in an approved animal biosafety level 3 laboratory. Serology and polymerase chain reaction (PCR) analysis were conducted to confirm that the civets had not been previously exposed to SARS-CoV. Animal experiments were conducted in accordance with the animal ethics guidelines and approved
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