A serosurvey of swine and free-living species on Iowa farms for antibodies against encephalomyocarditis virus

Swine and free-living nonporcine species from 20 Iowa swine farms were surveyed for antibodies against encephalomyocarditis virus (EMCV). The microtitration serum neutralization test was used, and antibody titers ≥ 1:8 were considered positive. The overall prevalence in 267 sows in various groups was 37.8% (range 20-86%). The prevalence in sows maintained in total confinement was significantly lower than in sows not maintained in total confinement (p = 0.01). Prevalence in sows was not associated with other herd characteristics or with abundance estimates of rats and/or mice.

Free-living animals tested included 74 domestic cats (Felis domestica), 203 house mice (Mus musculus), 15 mice of the genus Peromyscus, nine Norway rats (Rattus norvegicus), 34 opossums (Didelphis virginiana), 14 raccoons (Procyon lotor), and seven striped skunks (Mephitis mephitis). Of these, the only seropositive animals were two domestic cats.

The results of this study failed to implicate the free-living species surveyed as important reservoirs of EMCV for swine, and suggested that swine themselves and/or some species not included in this survey are the main reservoir of EMCV for swine in Iowa.

     

Pseudorabies virus antibodies in swine slaughtered in Iowa.

Sera from butcher swine (1,246 total) were evaluated qualitatively by the microimmunodiffusion test and quantitatively by the virus neutralization test for antibody to pseudorabies virus. Ten percent of the sera had antibody to pseudorabies virus. Follow-up contact with veterinarians whose clients included the farms from which the positive swine originated revealed that few feeder swine are vaccinated against pseudorabies and that most infections with pseudorabies virus are subclinical.     

猪源脑心肌炎病毒单克隆抗体研制与免疫学特性测定

利用纯化灭活的猪源脑心肌炎病毒(EMCV)免疫BALB/c小鼠,采用杂交瘤细胞技术,研制获得4株能稳定分泌抗EMCV抗体的杂交瘤细胞株,分别命名为1D1、2A2、2B6和4E2。经ELISA测定,细胞培养上清中抗体效价分别为1∶1 600,1∶6 400,1∶6 400和1∶3 200,小鼠腹水抗体效价分别为1∶1.63×106,1∶3.28×106,1∶1.13×106和1∶5.63×105。1D1和2A2单抗亚类为IgG1,2B6和4E2单抗亚类为IgG2b,轻链均为κ型。Western blot结果表明,1D1、2A2和4E2能特异性识别病毒的VP1蛋白,2B6能特异性识别病毒的VP2蛋白。间接免疫荧光试验证明,4株单抗具有良好的特异性,均能识别EMCV。本研究获得的4株特异性单抗,将为EMCV诊断方法的建立和病毒蛋白功能的研究奠定基础。     

Prevalence of coronavirus antibodies in Iowa swine.

Three hundred and forty-seven serum samples from 22 Iowa swine herds were screened for TGEV/PRCV neutralizing antibody. Ninety-one percent of the sera and all 22 herds were positive. These sera were then tested by the blocking ELISA test to distinguish TGEV and PRCV antibody. The ELISA test confirmed the high percentage of TGEV/PRCV positive sera. By the blocking ELISA test, 12 herds were PRCV positive, 6 herds were TGEV positive and 4 herds were mixed with sera either positive for TGEV or PRCV antibody. The results suggest a recent increase in TGEV/PRCV seroprevalence in Iowa swine most likely due to subclinical PRCV infections.     

Characterization of encephalomyocarditis virus isolated from aborted swine fetuses.

Characteristics of 2 encephalomyocarditis virus (EMCV) isolates (MN-25 and MN-30) recovered from aborted swine fetuses were examined along with 2 other swine isolates (NVSL-MDV and NVSL-PR) and a reference ATCC strain (VR-129). All 5 EMCV isolates were found to be serologically related by cross testing, using serum neutralization and fluorescent antibody assays. Hemagglutination (HA) properties of the 5 isolates were compared, using 5 diluents. The MN-25 and NVSL-MDV strains had HA activity with guinea pig RBC in all 5 diluents, whereas MN-30, NVSL-PR, and VR-129 had HA activity only in KCl-borate buffer. The HA ability with RBC of various animal species was examined, using KCl-borate diluent. All virus isolates had high HA titer (1:512 to 1:2,048) with guinea pig, rat, and horse RBC and lower HA titer (1:16 to 1:64) with sheep RBC. The MN-25 and NVSL-MDV isolates agglutinated dogs RBC, whereas MN-30, NVSL-PR, and VR-129 strains did not. Viral replication was evident in 8 of 10 cell lines tested, although infectivity titers of each virus varied by cell line used. Plaque-forming ability was similar for all 5 isolates, but plaque size was different by virus and cell culture used. Virus isolates were found to be stable after being heated at 56 C and subjected to a wide range of pH. A viral polypeptide pattern difference for all 5 isolates was not found by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of serologic methods for the detection of antibodies to encephalomyocarditis virus in swine fetal thoracic fluids.

Hemagglutination inhibition (HI) and agar gel immunodiffusion (AGID) tests were compared to the serum neutralization (SN) test to evaluate their ability to detect antibodies to encephalomyocarditis virus (EMCV). Swine fetal thoracic fluids of known EMCV SN antibody titers (200 samples greater than or equal to 1:2, 100 samples less than 1:2) were selected from a collection of field cases. The thoracic fluids were tested for EMCV antibodies by HI and AGID, and the results were compared to those of the SN test. Of 200 SN antibody-positive samples, 183 (91.5%) and 173 (86.5%) were positive in HI and AGID tests, respectively. Of 100 SN-negative samples, 81 (81%) and 94 (94%) were negative in HI and AGID tests, respectively. Agreement between the tests was analyzed by calculating Kappa values. The values were 0.73 between SN and HI tests and 0.77 between SN and AGID tests, indicating very good to excellent agreement for HI and AGID tests with the SN test. Of 200 SN-positive samples, 19 samples with low SN titers (1:2-1:16) were further tested by Western immunoblotting, and all were confirmed as positive. Interpretation of the present results suggests that both HI and AGID tests can be used as alternatives to the SN test.     

Seroprevalence of antibodies against bluetongue virus in animals imported to Poland from EU countries

The aim of this study was to determine the seroprevalence of BTV-specific antibodies in animals imported to Poland from EU countries after 15 June 2006. From 1 January 2007 to 22 January 2008, a total of 10719 samples of sera collected from cattle, goats and fallow deer were tested. Sera were screened using the highly sensitive and specific c-ELISA test and positive results were confirmed by the AGID assay. Out of 10719 sera, 30 (0.28% of the total number of samples) were found to be positive in both tests applied. All of 21 seropositive cattle specimens were imported to Poland from Germany whereas 9 seropositive fallow deer were of Dutch origin. In conclusion, it can be stated that because BTV situation in Europe is getting worse, implemented surveillance studies should be continued to monitor the actual BT status in Poland.     

珠海地区猪流感血清学的调查报告

2005年初至2007年上半年,对珠海市的不同猪场的猪流感的血清学普查,共检测1673份猪血清和56份人血清样品。结果表明,该市被检猪场流行的流感血清亚型是H1和H3,并以H1为主,未检测到H5、H9亚型抗体。总体来说,集约化猪场比个体(散户)猪场更容易受到感染,母猪的抗体阳性率及滴度明显比断奶猪和中大猪高。养猪场职工血清H1和H3阳性水平与其性别、年龄及与猪密切接触时间的长短没有直接关系。     

An immunoperoxidase monolayer assay for the detection of antibodies against swine influenza virus.

An immunoperoxidase monolayer assay (IPMA) has been developed to detect antibodies against swine influenza A virus (SIV) in pig sera. The test was evaluated by using sequential sera from pigs experimentally infected with H1N1 subtype of SIV. Two hundred field serum samples that had been examined by the hemagglutination inhibition (HI) test were also tested. Antibodies specific to SIV were detected as early as 3 days postinoculation (dpi) in the IPMA test as compared with 7 dpi by the HI test. Unlike HI, no serum treatment was required in the IPMA test. Regardless of the virus used in the test, IPMA detected antibodies to both H1N1 and H3N2 subtypes of SIV whereas HI detects antibodies against either H1N1 or H3N2, depending upon the virus used in the test. Results of this study indicate that IPMA is a useful test for screening of pig sera for SIV antibodies.     

Detection of antibodies against African swine fever virus using infected monolayers and monoclonal antibodies

Three monoclonal antibodies, specific for porcine IgG, IgM and IgA, were used to develop isotype-specific immunoperoxidase monolayer assays for the detection of antibodies against African swine fever virus. A mixture of anti-IgM and anti-IgG monoclonal antibodies was used in an assay designed for screening sera. This test was compared with a commercially available ELISA by using experimental sera and field sera obtained after an outbreak of African swine fever on two farms in the Netherlands in 1986. Although the ELISA was less sensitive than the immunoperoxidase monolayer assay on sera taken early after infection, the tests were equally useful for screening purposes. The isotype-specific assays gave epizootiological information about the stage of infection on the two farms.     

具有HI活性的H1亚型流感病毒特异性单克隆抗体的制备与应用

为了制备具有HI活性的HI亚型流感病毒特异性单克隆抗体(MAb),本研究以H1N1亚型猪流感病毒(SIV)株A/Swine/Guangdong/718/01(H1N1)为免疫原,免疫BALB/c小鼠,经常规细胞融合后,血凝抑制(HI)方法进行检测,融合细胞经稀释克隆纯化后,获得11株能稳定分泌抗血凝素特异性HI MAb的杂交瘤细胞株。鉴定表明,所获MAb与其他具有血凝活性的病毒以及其他14个HA亚型的流感病毒均不具有HI交叉反应,表明这11株MAb均具有良好的流感病毒亚型特异性。其中A6F、2BBF和2BB与其他H1亚型流感病毒分离株的HI试验证实我国不同地区分离株之间的抗原性存在一定差异。11株MAb对H1SIV抗原的HI试验结果显示其HI效价有明显差异。叠加实验表明这些MAb分别识别HA抗原的不同表位。间接免疫荧光试验表明,2BBF、8HB、1DH、7FC和2BB均可与2009年流行H1N1病毒A/California/04/2009HA抗原发生特异性反应。这些MAb特异性的研制为H1亚型流感病毒的疫情病原学快速诊断以及病毒抗原性变异的相关研究提供了物质基础。     

Ochratoxicosis in Iowa swine

Ochratoxicosis was diagnosed in 2 Iowa swine herds. In both cases, clinical signs consisted of increased urination and water consumption. Pale tan kidneys were found in affected pigs at necropsy. Histologic examination of renal tissue from an affected pig revealed diffuse tubular nephrosis and interstitial fibrosis. In both cases, ochratoxin A was detected by thin-layer chromatography in extracts of whole corn and ground complete feed. Ochratoxicosis was reproduced in swine with feed obtained in one of the cases.     

Seroprevalence of anti-Toxoplasma IgG in Canadian swine.

In 1990, 1443 sera randomly selected by computer generated random numbers from a bank of greater than 15,000 sera collected earlier from sows at slaughter in various geographic regions of Canada were tested for anti-Toxoplasma immunoglobin G (IgG) by the modified direct agglutination test. One hundred and thirty-six (9.4%) samples were positive for anti-Toxoplasma IgG while 36 (2.5%) of the sera gave borderline reactions. The highest prevalence of anti-Toxoplasma IgG was in swine originating in the Atlantic provinces and Ontario while the lowest was in swine from Manitoba and Saskatchewan.     

Production of monoclonal antibodies against swine fever virus and their use in laboratory diagnosis

Thirteen hybridoma cell lines which secrete monoclonal antibodies (MCAs) against swine fever virus (SFV) strain Brescia were produced. The hybrid cells resulted from fusion of P3X63-Ag8.653 myeloma cells with splenocytes of Balb/c mice which had been immunized with purified SFV. Screening of supernatant fluids was performed by an indirect enzyme-linked immunosorbent assay (ELISA) and an immunoperoxidase monolayer assay (IPMA). The IPMA, and an immunoperoxidase test (IPT) performed on cryostat sections, were used to characterize these MCAs on several pestivirus strains. All MCAs reacted to a varying degree with all but one of the SFV strains tested. None of the MCAs reacted with the bovine viral diarrhoea virus (BVDV) strains. Two MCAs are now used routinely in the differential diagnosis between infections with field strains of SFV, and the Chinese vaccine strain and BVDV strains.     

猪流感抗体间接ELISA检测方法的建立

猪流感病毒A／Swine／Fujian／668／2001（H3N2）株感染的鸡胚尿囊液，经差速离心后，再经蔗糖密度梯度离心，提纯、纯化的猪流感病毒经NP-40处理并反复冻融，作为猪流感间接ELISA抗原，确立了间接ELISA检测方法。对29份HI试验猪流感为阴性的血清进行了检测，经统计学分析，确定间接ELISA判定标准，被检血清OD490nm值≥0．20判定为阳性。该方法对猪瘟等11种猪疫病阳性血清无交叉反应，批内和批间重复试验的吸收变异系数分别在3．34％～8．12％和6．2％～9．04％之间。与HI的符合率达到92．8％，经卡方检验（P〈0．01）比HI试验敏感。为猪流感抗体检测提供了快速、准确、简便的方法。     

Seroprevalence of antibodies against equine arteritis virus in horses residing in the United States and imported horses

OBJECTIVE: To compare seroprevalence of antibodies against equine arteritis virus (EAV) in horses residing in the United States with that of imported horses. DESIGN: Serologic survey. SAMPLE POPULATION: Serum samples from 364 horses on 44 equine operations in California and 226 horses imported from various countries. PROCEDURE: Serum samples were collected from each imported horse and from up to 20 horses on each operation. For resident horses, the number of sampled horses on each operation was determined on the basis of the number of horses on the operation. Samples were tested for antibodies against EAV by use of a serum neutralization test. RESULTS: 1.9% of resident horses and 18.6% of imported horses were seropositive to EAV, including 16.1% of imported stallions. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that the EAV seroprevalence of horses residing in California is considerably lower than that of imported horses, including imported stallions.