Transplacental infection following exposure of gilts to porcine reproductive and respiratory syndrome virus at the onset of gestation

Twenty-five gilts without measurable porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) serum antibody titres were used for this experiment. All of them were randomly assigned to one of the treatment groups at the time of artificial insemination. Twelve gilts were exposed to PRRSV, of these, six were slaughtered on day 10 after exposure and constituted group A. The remaining six were slaughtered on day 20 after infection and constituted group C. Thirteen gilts were used as controls, six of these were slaughtered on day 10 after treatment and constituted group B. The remaining seven were slaughtered on day 20 after treatment and constituted group D. The infected gilts were inoculated with PRRSV intranasally and intravenously in the ear vein. They were observed for clinical signs of infection and the effects on conception and fertilization rates were studied, while the gilts and their embryos were tested for PRRSV and homologous antibodies. The infected animals developed signs of PRRS associated with anorexia and slight pyrexia. Infection was verified by reisolation of the virus from serum and other tissue samples and also by seroconversion. Ten out of 12 infected gilts and 10 out of 13 controls were pregnant at the time of slaughter and the ratio of embryos to corpora lutea was the same in both, infected and control groups (0.75). Therefore, infection with PRRSV at the onset of gestation did not appear to interfere with conception and fertilization rates and subsequent pregnancy. The PRRSV was not isolated from any of the embryos collected at day 10 postexposure, but was present in 20-day-old embryos of group C gilts. In this group, 60% of litters were infected prenatally, with 16% of embryos infected. The proportion of dead embryos was three times greater than in control group D (35.4% and 9.8%, respectively). The results of this report indicate that exposure of susceptible gilts to PRRSV at the onset of gestation has no significant effect on conception and fertilization rates. However, although infection does not appear to have any effect on the embryos before implantation, it can result in transplacental infection and embryo death.     

Eradication of porcine reproductive and respiratory syndrome virus by serum inoculation of na?ve gilts

The acclimatization program included exposure to serum and recovery. A continuous flow unit (nursery to finishing) from the same farm was selected as a potential source of porcine reproductive and respiratory syndrome virus (PRRSV). Negative gilts were inoculated 5 d after arrival by intramuscular injection of serum from selected animals. There was a significant reduction in seroprevalence in the sow herd 1 y after implementation of the gilt inoculation program (P < 0.05). At that time, all of the tested nursery pigs were negative for PRRSV. The fully segregated finisher population had a significant reduction in the frequency of PRRSV positive animals (P < 0.05) by enzyme-linked immunosorbent assay (ELISA), with all animals testing negative by the end of the study. However a persistent seroconversion was observed in the partially segregated finisher pigs (P > 0.05). In conclusion, the gilt serum inoculation program achieved sow herd stabilization, as defined by the production of negative weaned pigs and this resulted in the eradication of PRRSV in the fully segregated flow.     

Isolation of porcine reproductive and respiratory syndrome (PRRS) virus in a Danish swine herd and experimental infection of pregnant gilts with the virus

The first case of porcine reproductive and respiratory syndrome (PRRS) in Denmark was diagnosed in March 1992 by the detection of specific antibodies against PRRS virus in serum samples originating from sows in a herd located on the island of Als. Subsequently, PRRS virus was isolated from a 200-sow farrow-to-finish herd with clinical signs consistent with PRRS. The virus was isolated by inoculation of pleural fluid from a stillborn piglet onto porcine pulmonary alveolar macrophages. The isolate was identified as PRRS virus by staining with a specific antiserum. By electron microscopy, the virus particle was found to be spherical and enveloped, measuring 45–55 nm in diameter and containing a 30–35 nm nucleocapsid. Only minor antigenic differences were found between the Danish and a Dutch isolate. Following intranasal inoculation of 3 pregnant gilts with the Danish isolate transplacental infection was demonstrated by the re-isolation of PRRS virus from approximately 45% of the piglets from the experimentally infected gilts. However, the experimental infection produced no significant reproductive disorders or other clinical signs. At autopsy, histopathological examination revealed slight interstitial pneumonia in a few piglets.     

Infection dynamics of porcine reproductive and respiratory syndrome virus in a continuous-flow population of pigs also infected with Mycoplasma hyopneumoniae

Twenty-eight 10-week-old pigs were inoculated intratracheally with 1 x 10(5) colour-changing units/ml Mycoplasma hyopneumoniae strain 232, and another 32 pigs were not inoculated but were divided into 12 direct-contact pigs and 20 indirect-contact pigs. Thirty-five days later, the inoculated pigs were inoculated intranasally with 1 x 10(2.4) tcid50 of porcine reproductive and respiratory syndrome virus (PRRSV) strain mn 30-100. Viraemia, seroconversion and the transmission of PRRSV in the M hyopneumoniae-infected pigs were then assessed for four months. Three groups of 10 age-matched gilts were introduced as sentinels into the experimental barn on days 28, 56 and 84 after the PRRSV infection. The persistence of PRRSV was evaluated in both the experimentally and naturally infected pigs, which were slaughtered 120, 135 and 150 days after the infection. The period of viraemia and the extent of seroconversion were similar to those observed in studies of pigs infected only with PRRSV, suggesting that under the conditions of the study M hyopneumoniae did not affect these features of the disease. A delayed pattern in the seroconversion and proportion of pcr-positive pigs was observed in the direct and indirect contact groups, and the persistence of PRRSV in tissues was confirmed by pcr at 120 and 150 days after infection only in the directly inoculated pigs and not in the direct- or indirect-contact groups of pigs.     

Strain predominance following exposure of vaccinated and naive pregnant gilts to multiple strains of porcine reproductive and respiratory syndrome virus

Two studies were performed in order to test the relative ability of different strains of porcine reproductive and respiratory syndrome virus (PRRSV) to replicate and cross the placental barrier in pregnant gilts. Study 1 comprised 6 nonvaccinated gilts. Study 2 comprised 8 nonvaccinated gilts and 12 gilts that were vaccinated twice before conception. On, or about, gestation day 90 all gilts were simultaneously exposed to 20 field strains of PRRSV (all strains were distinguishable by restriction fragment length polymorphism (RFLP) patterns). Gilts of study 1 were euthanized on day 7 postpartum. Gilts of study 2 were euthanized on, or about, gestation day 111. All gilts, pigs, and fetuses were tested for the presence and type of strain of PRRSV. Of 128 samples shown to contain PRRSV, 118 contained a single strain, 4 contained 2 strains, and 2 contained a strain or strains for which the RFLP pattern was undecipherable. Only 8 of the 20 strains were isolated from nonvaccinated gilts and their litters. And only 2 of the 20 strains (notably 2 of the same strains isolated from nonvaccinated gilts and their litters), were isolated from vaccinated gilts and their litters. Moreover, 1 of the 2 strains accounted for most (31 of 37; 84%) of the isolates from the vaccinated group. Collectively these results indicate that strains differ in their ability to replicate in pregnant gilts and cross the placental barrier. And they suggest that maternal immunity, although sometimes insufficient to prevent transplacental infection, can exert additional selective pressure.     

Experimental inoculation of late term pregnant sows with a field isolate of porcine reproductive and respiratory syndrome vaccine-derived virus.

The use of a live attenuated porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in piglets has been associated with reproductive disorders in non-vaccinated sows. Vaccine-derived virus (VDV) has been isolated from foetuses, stillborn pigs, and dead piglets, indicating that the live vaccine spread from vaccinated piglets to non-vaccinated sows, and that the virus might be implicated in the severe reproductive problems observed. In the present study, one such VDV isolate was used to experimentally infect pregnant sows in the last trimester. The chosen isolate, which had more than 99.6% identity to the attenuated vaccine virus, originated from the lungs of a stillborn pig from a swine herd with a sudden high level of stillborn pigs and increased piglet mortality in the nursing period. Intranasal inoculation of sows with the virus isolate resulted in congenital infection, foetal death, and preweaning pig mortality. As such, the present study showed that vaccine-derived PRRSV can cause disease in swine consistent with PRRS.     

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…     

A review of porcine reproductive and respiratory syndrome virus in Dutch breeding herds: population dynamics and clinical relevance

Understanding the spread of porcine reproductive and respiratory syndrome virus (PRRSV) in pig populations is essential to the development of effective PRRS prevention and control strategies. Moreover, knowledge of the field dynamics of PRRSV in pigs will provide insights into the clinical relevance of PRRS, and will enable the targeting of interventions. This review of PRRSV includes discussion on the occurrence of outbreaks, the persistence of infection and the fade-out of infection in Dutch breeding herds. The dynamic character of PRRSV infections in endemically infected herds and the relevance of the disease under Dutch field conditions are also highlighted. Furthermore, several strategies aimed at controlling the spread of PRRSV are discussed.     

不同猪繁殖与呼吸综合征减毒活疫苗在猪体内的动态学及临床免疫效果比较

为了比较国内不同猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome,PRRS)弱毒疫苗的安全性和有效性,本研究将17头PRRSV抗原和抗体双阴性的28日龄仔猪随机分为4组,TJM-F92株弱毒疫苗组,JXA1-R株弱毒疫苗组,经典株弱毒疫苗(VR2332来源)组和注射生理盐水对照组。免疫后7,14,21,28d分别采血,ELISA抗体检测试剂盒检测PRRSV抗体,结果从免疫后14d起PRRSV抗体开始转阳,但各试验组的抗体水平无明显差异。对免疫后14d所采血样进行病毒分离,TJM-F92株免疫组未分离到PRRSV疫苗毒,JXA1-R株免疫组2头猪分离到PRRSV疫苗毒,经典株免疫组只1头猪分离到PRRSV疫苗毒。免疫后28d,所有猪接种PRRSV强毒株TJ株进行攻毒试验,观察各组猪的体温、临床症状;攻毒后21d,所有猪安乐死,剖检观察肺部病理变化。结果显示,TJM-F92株和JXA1-R株免疫组均未出现临床发病猪,经典株免疫组2头猪出现临床发病,对照组全部临床发病,死亡1头。经典株免疫组临床发病猪出现典型PRRSV感染肺部病理变化,而TJM-F92株和JXA1-R株免疫组均未发现典型PRRSV感染肺部病理变化。本研究证实TJM-F92株和JXA1-R株对高致病性PRRSV的免疫效果优于经典株弱毒疫苗,TJM-F92疫苗株与JXA1-R疫苗株相比,疫苗毒在体内带毒时间更短,安全性更高。     

Assessing the duration of persistence and shedding of porcine reproductive and respiratory syndrome virus in a large population of breeding-age gilts

Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus in the order Nidovirales, family Arteriviridae, genus Arterivirus. The virus induces a prolonged viremia, replicates in macrophages, and produces persistent infection. The purpose of this study was to determine if PRRSV could persist for 90 d or more in a large population of breeding-age gilts housed under environmental conditions typical of commercial swine production and to determine if experimentally infected gilts could shed virus to naïve sentinel gilts beyond 90 d postinfection. Using the intranasal route, we inoculated 120 PRRSV-naïve gilts, 4 mo of age, with 5 mL of cell culture fluid containing a total dose of 10^2.4 TCID₅₀ of a field isolate (MN-30100) of PRRSV. The index gilts were organized into 3 groups (A, B, and C), 40 gilts per group. To assess the dynamics of the experimental infection, a monitor group of 30 index gilts was blood-tested on days 0, 3, 7, 14, 30, 60, 90, 120, 150, and 180 postinfection. PRRSV viremia was detected with the polymerase chain reaction (PCR) on days 3, 7, and 14 and by virus isolation (VI) on days 7 and 14. PRRSV antibodies were detected from day 14 by enzyme-linked immunosorbent assay (ELISA). To assess shedding, 30 PRRSV-naïve sentinel gilts were commingled with the index gilts on day 90 postinfection and tested by PCR, VI, and ELISA every 15 d until 180 d postinfection; all samples were negative. To assess persistence, 40 index and 10 sentinel gilts were slaughtered at 120 (group A), 150 (group B), or 180 (group C) d postinfection. Evidence of PRRSV was not detected by PCR or VI in any tissue samples from the 120 index gilts. These results indicate that persistence and shedding of PRRSV are of short duration in breeding-age gilts.     

Experimental reproduction of porcine respiratory disease complex in pigs inoculated porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae and followed by inoculation with porcine circovirus type 2

The aim of this study was to reproduce severe pneumonic lesions, similar to those during naturally-occurring porcine respiratory disease complex, in pigs dually inoculated with porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae at 6 weeks of age, followed by inoculation with porcine circovirus type 2 at two weeks after. Time and sequence of infection with three pathogens mirror Asian field conditions. Microscopically, interstitial pneumonia and peribronchiolar lymphoid hyperplasia are considered the most characteristic lung lesions in infected pigs. The results of the present study demonstrate that inoculation of pigs with these three pathogens can lead to severe interstitial pneumonia with peribronchial or peribronchiolar lymphoid hyperplasia and fibrosis.     

Efficacy of vaccination with porcine reproductive and respiratory syndrome virus following challenges with field isolates in Japan

The objective of this study was to evaluate the influences of genetic and antigenic variations in field isolates of porcine reproductive and respiratory syndrome virus (PRRSV) on vaccine efficacy. Four-week-old pigs were vaccinated with a commercial modified live virus vaccine. Four weeks after vaccination, pigs in both the vaccinated group and the non-vaccinated group were challenged intranasally with 10(7) TCID(50) of PRRSV wt-11 (Experiment 1) or PRRSV wt-7 (Experiment 2). Based on genome sequencing of ORF5 and cross neutralization test results, PRRSV wt-11 is similar to the vaccine strain, whereas wt-7 is distinct from the vaccine strain. In the vaccinated challenged groups, clinical signs were less severe, the mean rate of weight gain was greater, and gross lung lesions were less severe when compared with the non-vaccinated challenged groups in both experiments. In Experiment 1, the virus was isolated from serum at 3 days post-challenge, and the mean virus titers in broncho-alveolar lavage fluids (BALF) and tissues were lower in pigs in the vaccinated challenged groups compared with those in the non-vaccinated challenged group. In Experiment 2, virus isolation from serum, BALF and tissues showed no significant differences between the groups. These results suggest that commercial PRRSV vaccine could be effective in reducing clinical disease following a challenge with field isolates of PRRSV. However, with regards to virological protection, the efficacy of the vaccine may be affected by the nature of the PRRSV isolates.     

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。     

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.     

猪繁殖与呼吸综合征病毒相关猪microRNA的初步筛选

本试验旨在初步筛选出与猪繁殖与呼吸综合征病毒(PRRSV)感染相关的猪microRNA(miRNA)。采用生物信息学方法预测可能与PRRSV 3′非编码区(3′utr)产生相互作用的猪miRNA,构建PRRSV 3′utr的双荧光素酶报告基因载体psi-CHECK-utr,同时合成预测到的miRNA(ssc-miR-323、ssc-miR-105-1)及其相应的抑制物,共转染猪脐静脉血管内皮细胞系(SUVEC),检测双荧光素酶活性。结果发现,ssc-miR-323与psiCHECK-utr共转染后,荧光比率略有升高,ssc-miR-105-1与psiCHECK-utr共转染后,荧光比率降低。初步判定ssc-miR-323对PRRSV 3′utr没有抑制作用,而ssc-miR-105-1对PRRSV3′utr有一定的抑制作用。     

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。