Evaluation of aerosol transmission of porcine reproductive and respiratory syndrome virus under controlled field conditions

The aim of this study was to determine whether porcine reproductive and respiratory syndrome virus (PRRSV) could be transmitted by aerosol under field conditions. A total of 210 five-month-old PRRSV-negative pigs were housed in a mechanically ventilated finishing facility containing 11 pens. Pen 1 contained 10 pigs (indirect contact controls) and pen 2 remained empty, providing a barrier of 2.5 m from the remaining pigs in pens 3 to 11. Fifteen or 16 of the pigs in each of pens 3 to 11 were infected experimentally with a field isolate of PRRSV and the other six or seven pigs served as direct contact controls. Five days after the pigs were infected, two trailers containing 10 five-week-old PRRSV-naive sentinel pigs were placed along each side of the building; one was placed 1 m from the exhaust fans on one side of the building, and the other was placed 30 m from the fans on the other side, and the sentinel pigs remained in the trailers for 72 hours. They were then moved to separate buildings on the same site, 30 and 80 m, respectively, from the infected barn, and their PRRSV status was monitored for 21 days. The direct and indirect contact control pigs became infected with PRRSV but the sentinel pigs did not.     

Experimental airborne transmission of porcine reproductive and respiratory syndrome virus and Bordetella bronchiseptica

Experiments were designed to determine if porcine reproductive and respiratory syndrome virus (PRRSV) or Bordetella bronchiseptica could be transmitted through indirect airborne contact. Three principal pigs were infected with PRRSV, B. bronchiseptica or both. Five days after the principal pigs were challenged, the three principal pigs and one direct-contact pig were placed into one isolation tent together, and three indirect-contact pigs were placed into another isolation tent which received its air supply from the first isolation tent. Airborne transmission of B. bronchiseptica occurred in 5/5 trials where B. bronchiseptica was the only agent used, and in 3/5 trials where the principal pigs were coinfected with both agents. Airborne transmission of PRRSV occurred in 4/5 trials where PRRSV was the only agent used, and in 2/5 trials where the principal pigs were coinfected with both agents. Thus, airborne transmission of both agents over short distances, such as within a barn, is probable.     

Evaluation of the aerosol transmission of a mixed infection of Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus

To evaluate the transmission of Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus (PRRSV) by aerosol as either a single or mixed infection, 28 pigs were inoculated intratracheally with M hyopneumoniae on day 0 and infected intranasally with PRRSV on day 35; they were housed together in a barn. To assess the aerosol transmission of M hyopneumoniae as a single infection, one trailer (A) containing 10 five-week-old sentinel pigs was placed along the south side of the infected barn (1 m from the fans) on day 28. To assess the mixed infection, two trailers (B and C), each containing 10 five-week-old sentinel pigs, were placed along each side of the barn on day 42. The sentinel pigs in the three trailers were exposed to the exhaust from the fans for seven days. No M hyopneumoniae infection was detected in the sentinel pigs in trailer A, but it was detected in the sentinel pigs in trailers B and C. No PRRSV was detected in any of the sentinel pigs.     

Further assessment of fomites and personnel as vehicles for the mechanical transport and transmission of porcine reproductive and respiratory syndrome virus

This study re-evaluated the role of fomites and personnel in the mechanical transport and transmission of porcine reproductive and respiratory syndrome virus (PRRSV) between pig populations. Swabs were collected from hands, boots, coveralls, and other fomites following contact with infected pigs and compared with identical samples collected in the absence of PRRSV exposure. Naïve pigs were provided contact with contaminated fomites/personnel and blood tested periodically post-exposure [positive exposure population (PEP)] and compared with populations that did not gain exposure via these routes [negative exposure population (NEP)]. The majority of swab samples from hands, coveralls, and boots from personnel and fomite samples (cable snare and bleeding equipment) following contact with the PRRSV-infected Source Population. Transmission of PRRSV to the PEP was observed (7/7) cases but not in the NEP. In conclusion, under the proper conditions, transport and transmission of PRRSV by fomites and personnel may occur between swine populations in the absence of intervention.     

Evaluation of the presence of porcine reproductive and respiratory syndrome virus in pig meat and experimental transmission following oral exposure

A study was performed to evaluate the presence of porcine reproductive and respiratory syndrome virus (PRRSV) in pig meat collected at slaughterhouses and its potential transmission to pigs via pig meat. A total of 1039 blood samples were collected from pigs upon their arrival at the abattoir. The following day, meat samples (n = 1027) were collected from the carcasses of these same pigs. Samples originated from 2 Canadian slaughterhouses, 1 situated in the province of Quebec and the other situated in the province of Manitoba. Serum samples were tested for antibodies to PRRSV and both serum and meat samples were also tested for PRRSV nucleic acid by polymerase chain reaction (PCR). Seropositivity to PRRSV for all serum samples was 74.3%. Furthermore 45 (4.3%) of the total serum samples and 19 (1.9%) of the 1027 meat samples were positive for PRRSV by PCR. Sequence analysis of open reading frame (ORF) 5 performed on 15 of the 19 PRRSV strains identified in pig meat indicated that 9 were field strains and 6 were vaccine-like (98% to 99.7% nucleotide homology with the Ingelvac RespPRRS/Repro vaccine). One of these 6 strains presented an intermediate 2-6-2 restriction fragment length polymorphism (RFLP) cut pattern and the others showed the characteristic 2-5-2 RFLP pattern of the vaccine strain. All strains sequenced were determined to be North American strains. In only 1 of the 19 PRRSV-positive meat samples could PRRSV be isolated. To test the potential infectivity of meat samples containing residual PRRSV, 11 of the PCR-positive meat samples (weighing 1.05 to 1.8 kg) were each used in feeding experiments of 2 PRRSV antibody-negative specific pathogen-free pigs of 9 wk of age. Samples were cut into several pieces and fed to each pair of pigs on 2 consecutive days. Each pig pair was housed in a separate cubicle and serum samples were collected at -7, 0, 7, 14, and 20 to 21 days post exposure. Seven pig pairs were found to be infected by PRRSV following ingestion of meat samples, including meat samples containing vaccine-like virus, as judged by the demonstration of PRRSV antibodies and/or PRRSV nucleic acid in the serum. In summary, the present study indicated that low residual quantities of PRRSV may be found in a small percentage of pig meat collected at slaugtherhouses. Furthermore, when this meat was fed raw to pigs in the experimental setting designed, pigs could be infected by PRRSV.     

猪繁殖与呼吸综合征病毒的研究进展

猪繁殖与呼吸综合征 (Porcine reproductiveand respiratory syndrome,PRRS)是 1 987年发现的一种新的猪病 ,它主要是引起母猪的繁殖障碍 ,如流产、早产、死胎、木乃伊胎和呼吸道症状的一种由猪繁殖与呼吸综合征病毒 (PRRSV)引起的接触性传染病。此病最初在美国首先被发现 ,随后相继在世界各国报道 ,我国台湾省也有疫情的报道。 1 995年我国首次从进口猪中检出 PRRS阳性猪 ,并从中分离到 PRRSV。 1 996年郭宝清等首次从国内 PRRS血清阳性猪群中分离到 PRRSV,从而证实了我国也有此病的流行[1] 。由于 PRRS主要是引起母猪的繁殖…     

Oral transmission of porcine reproductive and respiratory syndrome virus by muscle of experimentally infected pigs

The current study was performed to determine if porcine reproductive and respiratory syndrome virus (PRRSV) could be transmitted to pigs by feeding muscle tissue obtained from recently infected pigs. Muscle obtained from pigs infected with either a European strain (EU donor pigs) or American strain (US donor pigs) of PRRSV was fed to PRRSV-free receiver pigs. The donor pigs were slaughtered 11 days post-infection (dpi). PRRSV was detected by conventional virus isolation in muscle at 11 dpi from 7 of 12 EU donor pigs and 5 of 12 US donor pigs. In contrast to conventional virus isolation, all muscle samples from infected pigs were positive for viral nucleic acid by PCR, except for muscle from one animal infected with the American strain of PRRSV. Five hundred grams of raw semimembranosus muscle from each of the donor pigs was fed over a 2 days period (250 g per day) to each of two receiver pigs (48 receiver pigs). The receiver pigs were housed separately in five groups. One of the five groups was fed muscle obtained from US donor pigs that was also spiked with the American strain of PRRSV. Sentinel pigs were placed in-contact with the group of receiver pigs fed spiked muscle. All receiver pigs became viraemic by 6 days post-feeding (dpf). There was evidence of horizontal transmission with sentinel pigs, in-contact with receiver pigs, becoming viraemic. The study demonstrates that PRRSV could be infectious through the oral route via the feeding of meat obtained from recently infected pigs.     

猪繁殖与呼吸综合征病毒的分离与鉴定

从广东地区发病猪场的病料中,分离到1株致Marc-145细胞病变的病毒ShB6。扩增猪繁殖与呼吸综合征病毒(PRRSV)主要结构蛋白基因ORF2-ORF7并进行序列分析,结果表明,该分离株与国内PRRSV分离株HB-1(sh)/2002的同源性为96.9%;与ATCC VR-2332株的同源性为91%;而与Lelystad株的同源性仅为59.8%;用美洲型PRRSV单抗进行免疫组化染色,结果显示在细胞病变处呈现明显的阳性着色(为棕黄色)。综合可见,所分离的病毒为美洲型PRRSV。     

高致病性猪繁殖与呼吸综合征病毒(PRRSV)气溶胶的发生与传播机制

为深入认识猪繁殖与呼吸综合征病毒(PRRSV)气溶胶的发生与传播机制,将35日龄无PRRSV抗体仔猪预饲1周后分为4组(PRSSV攻毒组、直接接触组、间接接触组和阴性对照组),饲养于正负压隔离器中。2个隔离器置于不同室内并通过管道相连,采用AGI-30收集器收集其空气样品。然后,将其接种Marc-145细胞,再用RT-PCR检测病毒;并检测血常规和抗体变化。结果发现,攻毒组于攻毒后4d开始形成气溶胶,并持续到试验结束。气溶胶高峰出现在攻毒后15d。气溶胶传播的间接接触组从临床症状、病理变化、提取病理组织核酸和抗体检测等多个方面都证明其感染了PRRSV,并经鼻腔向外排毒;直接接触组和间接接触组几乎同时感染PRRSV。试验表明,PRRSV不仅能形成气溶胶并且能够通过其迅速感染临近猪群。     

猪繁殖与呼吸综合征病毒分子生物学研究进展

猪繁殖与呼吸综合征病毒(PRRSV)是引起猪繁殖与呼吸综合征的病原体,本文对PRRSV的分子生物学研究进展进行了综述,主要包括PRRSV的基因组结构、病毒蛋白及其功能、抗原变异等,旨在为诊断技术、免疫机理研究、疫苗设计与疫病防制提供借鉴.     

Survival of porcine reproductive and respiratory syndrome virus in houseflies

The objectives of the study were to determine the duration of porcine reproductive and respiratory syndrome virus (PRRSV) survival in houseflies (Musca domestica Linnaeus) following feeding on an infected pig, and to determine whether the virus was present on the exterior surface or within the internal viscera of the fly. A total of 210 laboratory-colonized houseflies were allowed to feed to repletion on a pig, experimentally infected with PRRSV on day 7 postinoculation, and then maintained alive under laboratory conditions (27°C). Two subsets (A and B) of 30 flies were collected at each of the following sampling points; 0, 6, and 12 hours post feeding (pf). Subset A contained an extra group of 30 flies collected at 24 hours pf due to the availability of extra flies. Flies in subset A were processed as whole fly homogenates, while the exterior surface washes and digestive organs were collected from flies in subset B. Whole fly homogenates, collected at 0, 6, and 12 hours pf, were positive by both polymerase chain reaction (PCR) and swine bioassay. Digestive organs, collected at 0 and 12 hours pf, were positive by PCR and swine bioassay. The PRRSV RNA was detected by PCR from the exterior surface wash of subset B flies collected at 0, 6, and 12 hours pf; however, only the subset collected at 0 hour pf was swine bioassay-positive. This study indicates that infectious PRRSV can survive within the intestinal tract of houseflies for up to 12 hours following feeding on an infected pig, but only for a short period on the exterior surface of the flies.     

Evaluation of the role of mallard ducks as vectors of porcine reproductive and respiratory syndrome virus

To assess the transmission of porcine reproductive and respiratory syndrome virus (PRRSV) from pigs to mallard ducks, 10 adult (one-year-old) female mallard ducks were housed with pigs infected experimentally with PRRSV, and allowed to be in close contact with them for 21 days. To evaluate the transmission of PRRSV from mallard ducks to pigs, two adult ducks were inoculated orally with PRRSV (total dose 10(6.0) TCID50) and allowed to drink PRRsv-infected water; 24 hours later, two four-week-old PRRsv-naive sentinel pigs were housed in pens below the cages housing the ducks for 14 days. In both experiments, cloacal and faecal samples were collected three times a week from the ducks and tested by PCR, virus isolation and a pig bioassay. Blood samples from the pigs were tested by ELISA, PCR and virus isolation. Sera from the ducks were tested by serum neutralisation. The ducks were examined postmortem and selected tissues were tested by PCR, virus isolation, histopathology and pig bioassay. In both experiments all the cloacal swabs, faecal samples, tissues and sera from the ducks were negative by all the tests. The sera from the pigs in the first experiment were PCR positive at three, seven, 14 and 21 days after infection and ELISA positive at 14 and 21 days. Sera from the pigs in the second experiment were negative by all the tests. The virus was isolated from the oral inoculum and the drinking water provided for the ducks in the second experiment. Under the conditions of this study, it was not possible to demonstrate the transmission of PRRSV either from the pigs to the ducks or from the ducks to the pigs.     

乳猪"高热症"猪繁殖与呼吸综合征病毒的分离与致病性分析

本研究针对疑似高热症疫情的猪场,从分离病原的致病特性及基因特征,对高热症的病因进行了探索。以RT—PCR／PCR从临床病料中检测出猪繁殖与呼吸综合征病毒（PRRSV）、猪圆环病毒2型（PCV2）、猪瘟病毒（CSFV）核酸,利用Marc-145细胞从病料中分离到PRRSV（XS070425毒株）。测序结果显示,XS070425毒株Nsp2基因无高致病性PRRSV Nsp2基因533～561位连续29个氨基酸“RPVTPLSEPIPVPAPRRKFQQVKRLSSAA”的缺失;但遗传进化分析显示,XS070425毒株Nsp2、ORF5基因与近期我国报道的高致病性PRRSV-HuN4和JXA1有较高的同源性,达95％～96％。致病性试验显示,PRRSV-XS070425原始病料悬液和Marc-145细胞分离物接种健康猪后,病毒血症可持续26d,并出现典型的PRRS临床症状和体温升高,但不致死感染猪。这些结果说明,无NSP2蛋白29个氨基酸残基缺失的PRRSV也是猪高热症的重要病原。     

猪繁殖与呼吸综合征病毒BJ-4株ORF3基因的原核表达

通过 PCR方法从重组质粒 p GEM- ORF3扩增得到缺失 N端疏水序列的基因片段 d ORF3(deleting ORF3)。将d ORF3克隆至原核高效表达载体 p GEX- 4 T- 2 ,在 E.coli BL 2 1细胞中成功表达了猪繁殖与呼吸综合征病毒 (PRRSV)重组蛋白GST- d ORF3,表达产物以包涵体的形式存在 ,表达量为 30 .6 % ,Western- Blot结果表明重组蛋白可被 PRRSV阳性血清所识别。表达的重组蛋白为进一步研究 PRRS病毒次要结构蛋白 GP3的免疫特性和功能奠定了基础     

猪繁殖与呼吸综合征病毒BJ-4杉ORF3基因的原核表达

通过PCR方法从重组质粒pGEM-ORF3扩增得到缺失N端疏水序列的基因片段dORF3(deleting ORF3)。将dORF3克隆至原核高效表达载体pGEX-4T-2，在E.coli BL21细胞中成功表达了猪繁殖与呼吸综合征病毒(PRRSV)重组蛋白GST-dORF3，表达产物以包涵体的形式存在，表达量为30．6％，Western-Blot结果表明重组蛋白可被PRRSV阳性血清所识别。表达的重组蛋白为进一步研究PRRS病毒次要结构蛋白GP3的免疫特性和功能奠定了基础。     

Failure of porcine reproductive and respiratory syndrome virus to replicate in porcine endothelial cell cultures.

Clinical, gross, and microscopic pathologic and immunohistochemical findings in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) suggest that PRRSV may replicate in endothelial cells. Endothelial cell cultures from porcine aorta and pulmonary artery were tested for susceptibility to various strains of PRRSV. Cultures were identified as endothelium by light microscopy and immunohistochemical staining for P-selectin and von Willebrand factor. Five strains of PRRSV, i.e., the National Veterinary Services Laboratories, Ames, IA PRRSV strain 130-PDV and 4 field strains isolated from pneumonic lungs, failed to replicate in these porcine large-vessel endothelial cell cultures.