成都市猫泛白细胞减少症病毒、星状病毒和肠道冠状病毒的分子流行病学调查
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摘要
为了解成都市区猫泛白细胞减少症病毒(FPV)、猫星状病毒(FAstV)和猫肠道冠状病毒(FECoV)的分子流行情况,本试验采集了315份家养猫和34份社区流浪猫的肛门拭子,含228份临床发病猫样本和121份临床健康猫样本,采用PCR方法对所有样本进行病毒核酸检测。同时,根据动物年龄、生活状态和免疫情况等因素进行病原流行分析,并根据病毒基因片段序列构建遗传进化树。结果显示,FPV、FAstV和FECoV的检出率分别为38.4%(134/349)、23.2%(81/349)和19.5%(68/349)。在混合感染方面,以FPV/FAstV(9.5%,33/349)、FAstV/FECoV(4.6%,16/349)和FPV/FECoV(4.3%,15/349)双重感染为主。基于部分基因序列的分析结果显示,样本间FPV VP2基因序列的同源性为95.1%～100%,发病猫毒株分为两个分支,其中一支与葡萄牙毒株(GenBank登录号:KT240136)和加拿大毒株(GenBank登录号:MF069445)遗传关系较近;另一支与中国的CPV毒株(GenBank登录号:KY937664)关系较近。样本间FAstV ORF1b基因序列同源性为90.9%～99.7%,发病猫的毒株的两个分支中一支与葡萄牙毒株(GenBank登录号:KF374704)和美国毒株(GenBank登录号:KM017743)关系较近;另一分支与中国香港毒株(GenBank登录号:KF499111)关系紧密。样本间FECoV的N基因序列同源性范围为89.3%～100%,与美国毒株(GenBank登录号:FJ938059)、意大利毒株(GenBank登录号:GU017092/GU017107)和中国毒株(GenBank登录号:KT852997)关系密切。调查结果表明,成都市区猫携带病毒性腹泻病原的情况较为普遍,部分猫存在多病原混合感染,且病毒基因序列存在明显变异,增加了病原流行及带毒猫发病的风险,需积极地进行临床防控。
To understand the molecular prevalence of feline panleukopenia virus(FPV), feline astrovirus(FAstV) and feline enteric coronavirus(FEcoV) in Chengdu, this experiment collected 315 domestic cats and 34 community stray cats anal swabs containing 228 clinically infected cat samples and 121 clinical healthy cat samples. Viral nucleic acids in all samples were detected by PCR. At the same time, pathogen epidemiology analysis was carried out according to factors such as animal age, living state and immune status, and genetic evolution trees were constructed based on the sequences of viral gene fragments. The results showed that the detection rates of FPV, FAstV and FECoV were 38.4%(134/349), 23.2%(81/349) and 19.5%(68/349), respectively. In the case of mixed infection, FPV/FAstV(9.5%, 33/349), FAstV/FECoV(4.6%, 16/349) and FPV/FECoV(4.3%, 15/349) were the main infections. Based on the analysis of partial gene sequences, the homology of the FPV VP2 gene sequence between samples was 95.1% to 100%. The sick cat strains were divided into two branches, one of which was closely related to the Portuguese strain(GenBank accession No.: KT240136) and the Canadian strain(GenBank accession No.: MF069445), and the other was closely related to the Chinese CPV strain(GenBank accession No.: KY937664).The sequence homology of FAstV ORF1b gene between samples was 90.9% to 99.7%. One of the two branches of the sick cat strain was closely related to the Portuguese strain(GenBank accession No.: KF374704) and the American strain(GenBank accession No.: KM017743). The other branch was closely related to the Hong Kong strain of China(GenBank accession No.: KF499111).The N gene sequence homology of FECoV between samples was 89.3% to 100%, all the FECoV strains sequences were in a branch showing high homology to American(GenBank accession No.:FJ938059),Italy(GenBank accession No.:GU017092/GU017107) and Chinese strain(GenBank accession No.:KT852997).The survey results showed that the situation of cats carrying viral diarrhea in Chengdu was more common, some cats had multiple pathogen mixed infections, and the viral gene sequence had significant variation, increasing the risk of pathogen epidemics and the incidence of poisoned cats. It was necessary to actively carry out clinical prevention and control.
To understand the molecular prevalence of feline panleukopenia virus(FPV), feline astrovirus(FAstV) and feline enteric coronavirus(FEcoV) in Chengdu, this experiment collected 315 domestic cats and 34 community stray cats anal swabs containing 228 clinically infected cat samples and 121 clinical healthy cat samples. Viral nucleic acids in all samples were detected by PCR. At the same time, pathogen epidemiology analysis was carried out according to factors such as animal age, living state and immune status, and genetic evolution trees were constructed based on the sequences of viral gene fragments. The results showed that the detection rates of FPV, FAstV and FECoV were 38.4%(134/349), 23.2%(81/349) and 19.5%(68/349), respectively. In the case of mixed infection, FPV/FAstV(9.5%, 33/349), FAstV/FECoV(4.6%, 16/349) and FPV/FECoV(4.3%, 15/349) were the main infections. Based on the analysis of partial gene sequences, the homology of the FPV VP2 gene sequence between samples was 95.1% to 100%. The sick cat strains were divided into two branches, one of which was closely related to the Portuguese strain(GenBank accession No.: KT240136) and the Canadian strain(GenBank accession No.: MF069445), and the other was closely related to the Chinese CPV strain(GenBank accession No.: KY937664).The sequence homology of FAstV ORF1b gene between samples was 90.9% to 99.7%. One of the two branches of the sick cat strain was closely related to the Portuguese strain(GenBank accession No.: KF374704) and the American strain(GenBank accession No.: KM017743). The other branch was closely related to the Hong Kong strain of China(GenBank accession No.: KF499111).The N gene sequence homology of FECoV between samples was 89.3% to 100%, all the FECoV strains sequences were in a branch showing high homology to American(GenBank accession No.:FJ938059),Italy(GenBank accession No.:GU017092/GU017107) and Chinese strain(GenBank accession No.:KT852997).The survey results showed that the situation of cats carrying viral diarrhea in Chengdu was more common, some cats had multiple pathogen mixed infections, and the viral gene sequence had significant variation, increasing the risk of pathogen epidemics and the incidence of poisoned cats. It was necessary to actively carry out clinical prevention and control.
引文
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[23] FRANZO G,TUCCIARONE C M,CECCHINATO M,et al.Canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV) codon bias analysis reveals a progressive adaptation to the new niche after the host jump[J].Molecular Phylogenetics & Evolution,2017,114:82-92.
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[25] RICE M,WILKS C R,JONES B R,et al.Detection of astrovirus in the faeces of cats with diarrhoea[J].New Zealand Veterinary Journal,1993,41(2):96-97.
[26] ZHANG W,LI L,DENG X,et al.Faecal virome of cats in an animal shelter[J].Journal of General Virology,2014,95(11):2553-2564.
[27] FOLEY J E,POLAND A,CARLSON J,et al.Patterns of feline coronavirus infection and fecal shedding from cats in multiple-cat environments[J].Journal of the American Veterinary Medical Association,1997,210(9):1307-1312.
[28] MARA B,AMBRA B,ALESSANDRA S,et al.Sequence analysis of the nucleocapsid gene of feline coronaviruses circulating in Italy[J].New Microbiologica Official Journal of the Italian Society for Medical Virology,2010,33(4):387-392.
[29] 刘秋瑾,孙东波,王欣宇,等.2015-2016年黑龙江三个地区猫冠状病毒N基因分子流行病学调查[J].黑龙江畜牧兽医,2018,15:130-134.LIU Q J,SUN D B,WANG X Y,et al.Molecular epidemic investigation of feline coronavirus N gene in three Heilongjiang districts in 2015-2016[J].Heilongjiang Animal Science and Veterinary Medicine,2018,15:130-134.(in Chinese)
[30] TEKELIOGLU B K,BERRIATUA E,TURAN N,et al.A retrospective clinical and epidemiological study on feline coronavirus (FCoV) in cats in Istanbul,Turkey[J].Preventive Veterinary Medicine,2015,119:41-47.
[2] PARIS A K,WILLS S,BALZER H,et al.Enteropathogen co-infection in UK cats with diarrhoea[J].BMC Veterinary Research,2014,10:13.
[3] MIRANDA C,VIEIRA M J,SILVA E,et al.Genetic analysis of feline panleukopenia virus full length VP2 gene in domestic cats between 2006-2008 and 2012-2014,Portugal[J].Transboundary & Emerging Diseases,2017,64(4):1178-1183.
[4] 温肖会,邱杰,吕殿红,等.广州市部分区域猫泛白细胞减少症流行现状调查[J].安徽农业科学,2016,44(17):120-122.WEN X H,QIU J,LYU D H,et al.Epidemiological survey of feline panleukopenia virus in partial region of Guangzhou city[J].Journal of Anhui Agricultural Sciences,2016,44(17):120-122.(in Chinese)
[5] AWARD R A,KHALIL W K B,ATTALLAH A G.Epidemiology and diagnosis of feline panleukopenia virus in Egypt:Clinical and molecular diagnosis in cats[J].Veterinary World,2018,11(5):578-584.
[6] FEI F D,YONG F Z,JIAN L W,et al.Molecular characterization of feline panleukopenia virus isolated from mink and its pathogenesis in mink[J].Veterinary Microbiology,2017,205:92-98.
[7] WANG X,LI T,LIU H,et al.Recombinant feline parvovirus infection of immunized tigers in central China[J].Emerging Microbes & Infections,2017,6(6):e42.
[8] LAU S K,WOO P C,YIP C C,et al.Complete genome sequence of a novel feline astrovirus from a domestic cat in Hong Kong[J].Genome Announcements,2013,1(5):e708-e713.
[9] LAWLER P E,COOK K A,WILLIAMS H G,et al.Determination of the diversity of astroviruses in feces from cats in Florida[J].Journal of Veterinary Diagnostic Investigation,2018,30(2):275-279.
[10] MOSCHIDOU P,MARTELLA V,LORUSSO E,et al.Mixed infection by feline astrovirus and feline panleukopenia virus in a domestic cat with gastroenteritis and panleukopenia[J].Journal of Veterinary Diagnostic Investigation,2011,23(3):581-584.
[11] CHO Y Y,LIM S I,KIM Y K,et al.Molecular characterisation and phylogenetic analysis of feline astrovirus in Korean cats[J].Journal of Feline Medicine & Surgery,2014,16(8):679-683.
[12] SHU S Y,JIANG T N,HUA L W,et al.Molecular characterization of feline astrovirus in domestic cats from Northeast China[J].PLoS One,2018,13(10):e205441.
[13] OGUMA K,OHNO M,YOSHIDA M,et al.Mutation of the S and 3c genes in genomes of feline coronaviruses[J].Journal of Veterinary Medical Science,2018,80(7):1094-1100.
[14] VOGEL L,VAN DER LUBBEN M,LINTELO E G,et al.Pathogenic characteristics of persistent feline enteric coronavirus infection in cats[J].Veterinary Research,2010,41(5):571-584.
[15] VENNEMA H,POLLAND A,FOLEY J,et al.Feline infectious peritonitis viruses arise by mutation from endemic feline enteric coronaviruses[J].Virology,1998,243(1):150-157.
[16] AN D J,JEOUNG H Y,JEONG W,et al.Prevalence of Korean cats with natural feline coronavirus infections[J].Virology Journal,2011,8:455.
[17] 刘碧涛,任常宝,张晓战,等.猫细小病毒的分离与鉴定[J].动物医学进展,2013,34(9):124-127.LIU B T,REN C B,ZHANG X Z,et al.Separation and isolation of feline panleukopenia virus[J].Progress in Veterinary Medicine,2013,34(9):124-127.(in Chinese)
[18] AKKAN H A,KARACA M.Studies on the seroprevalence,age,and gender on the distribution of feline coronavirus in Van cats kept in a multiple-cat environment[J].Bulletin-Veterinary Institute in Pulawy,2009,53(2):183-186.
[19] MELI M,KIOAR A,MULLER C,et al.High viral loads despite absence of clinical and pathological findings in cats experimentally infected with feline coronavirus (FCoV) type Ⅰ and in naturally FCoV-infected cats[J].Journal of Feline Medicine & Surgery,2004,6(2):69-81.
[20] MOCHIZUKI M,OSAWA N,ISHIDA T.Feline coronavirus participation in diarrhea of cats[J].Journal of Veterinary Medical Science,1999,61(9):1071-1073.
[21] AN D J,JEONG W,JEOUNG H Y,et al.Phylogenetic analysis of feline panleukopenia virus (FPLV) strains in Korean cats[J].Research in Veterinary Science,2011,90(1):163-167.
[22] AHMED N,RIAZI A,ZUBAIR Z,et al.Molecular analysis of partial VP-2 gene amplified from rectal swab samples of diarrheic dogs in Pakistan confirms the circulation of canine parvovirus genetic variant CPV-2a and detects sequences of feline panleukopenia virus (FPV)[J].Virology Journal,2018,15(1):45.
[23] FRANZO G,TUCCIARONE C M,CECCHINATO M,et al.Canine parvovirus type 2 (CPV-2) and feline panleukopenia virus (FPV) codon bias analysis reveals a progressive adaptation to the new niche after the host jump[J].Molecular Phylogenetics & Evolution,2017,114:82-92.
[24] MENDE K,STUETZER B,SAUTER L C,et al.Prevalence of antibodies against feline panleukopenia virus in client-owned cats in Southern Germany[J].Veterinary Journal,2014,199(3):419-423.
[25] RICE M,WILKS C R,JONES B R,et al.Detection of astrovirus in the faeces of cats with diarrhoea[J].New Zealand Veterinary Journal,1993,41(2):96-97.
[26] ZHANG W,LI L,DENG X,et al.Faecal virome of cats in an animal shelter[J].Journal of General Virology,2014,95(11):2553-2564.
[27] FOLEY J E,POLAND A,CARLSON J,et al.Patterns of feline coronavirus infection and fecal shedding from cats in multiple-cat environments[J].Journal of the American Veterinary Medical Association,1997,210(9):1307-1312.
[28] MARA B,AMBRA B,ALESSANDRA S,et al.Sequence analysis of the nucleocapsid gene of feline coronaviruses circulating in Italy[J].New Microbiologica Official Journal of the Italian Society for Medical Virology,2010,33(4):387-392.
[29] 刘秋瑾,孙东波,王欣宇,等.2015-2016年黑龙江三个地区猫冠状病毒N基因分子流行病学调查[J].黑龙江畜牧兽医,2018,15:130-134.LIU Q J,SUN D B,WANG X Y,et al.Molecular epidemic investigation of feline coronavirus N gene in three Heilongjiang districts in 2015-2016[J].Heilongjiang Animal Science and Veterinary Medicine,2018,15:130-134.(in Chinese)
[30] TEKELIOGLU B K,BERRIATUA E,TURAN N,et al.A retrospective clinical and epidemiological study on feline coronavirus (FCoV) in cats in Istanbul,Turkey[J].Preventive Veterinary Medicine,2015,119:41-47.