Acute phase response in porcine reproductive and respiratory syndrome virus infection

This study was focused on the changes observed in the serum concentration of haptoglobin (Hp), C-reactive protein (CRP), serum amyloid A (SAA) and Pig-major acute protein (Pig-MAP), during experimental porcine reproductive and respiratory syndrome virus (PRRSV) infection and in their relationship with the expression of interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α). Hp and Pig-MAP serum levels were increased at 10 dpi, but CRP and SAA showed a delayed and highly variable increase. All three proinflammatory cytokines were poorly expressed, and only a mild increase in IL-1β was observed at 7 dpi. The increased expression of Hp coincided with the light enhancement observed in both IL-6 and TNF-α, and might be related with an increased expression of IL-10. The low expression of TNF-α might point to a possible mechanism of viral evasion of host-immune response. This issue and the delayed expression of CRP and SAA should be taken into account in future studies about modulation of the immune response by PRRSV infection.     

Pathogenesis of in utero infection in porcine fetuses with porcine reproductive and respiratory syndrome virus.

Porcine fetuses were exposed in utero to porcine reproductive and respiratory syndrome virus (PRRSV) at stages of gestation ranging from 34 to 85 days and examined 17 to 31 days later to determine the effect of gestational age on fetal susceptibility. For each of the 8 litters tested during the study, all of the fetuses of 1 horn of the uterus were exposed to virus by intraamniotic injection; those of the other horn were exposed similarly to a sham inoculum that consisted of sterile cell culture medium. Viral infectivity titers associated with fetal tissues collected at necropsy indicated that, regardless of gestational age, the virus had replicated in fetuses exposed intraamniotically. In addition, virus had also spread and replicated in sham-inoculated littermates in 3 litters. On the basis of these findings it appears that there may be little or no temporal difference in fetal susceptibility to infection with PRRSV. If so, the lack of early fetal death as a commonly recognized feature of naturally occurring cases of PRRS may be due to a greater resistance of early gestational fetuses to the lethal effects of PRRSV, as suggested by this study, and/or a greater likelihood of transplacental infection during late gestation.     

Effects on boar semen quality after infection with porcine reproductive and respiratory syndrome virus: a case report

The effect of porcine reproductive and respiratory syndrome virus (PRRSV) on semen quality was examined in a group of 11 spontaneously infected boars in a commercial boar stud. Semen samples were collected 4 weeks prior to 4 weeks post-infection (wpi). Infection with PRRSV of the European genotype subtype 1 (EU-1) was verified by specific quantitative real-time polymerase chain reaction (RT-PCR) in 36% of the serum samples. All boars seroconverted before 4 wpi and remained in normal condition throughout the study. Comparison of the percentage of morphologically intact spermatozoa revealed an increase of acrosome-defective spermatozoa (P = 0.012) between −4 and 4 wpi. Significant deleterious effects on semen quality were detected for membrane integrity when semen had been stored for 2 days after sampling. Analysis of sperm subpopulations in a thermoresistance test on day 7 after sampling revealed alterations in the percentage of circular, progressively motile spermatozoa (P = 0.013), in the percentage of non-linear, progressively motile spermatozoa (P = 0.01), and on the amplitude of lateral sperm head displacement (P = 0.047). There was no difference in the incidence of mitochondrially active spermatozoa (P = 0.075). Investigation of routine production data between pre- and post-infection status showed no differences on ejaculate volume (P = 0.417), sperm concentration (P = 0.788), and percentage of motile spermatozoa (P = 0.321). This case report provides insights into a potential control strategy for PRRSV outbreaks in boar studs.     

Persistence of porcine reproductive and respiratory syndrome virus infection in a swine operation.

A herd of Quebec seedstock pigs experienced in early 1992 a typical outbreak of porcine reproductive and respiratory syndrome (PRRS) associated with lesions of interstitial, proliferative and necrotizing pneumonia in weaned piglets. The nature of the infection was confirmed by serology using indirect immunofluorescence (IIF) and virus isolation in primary cultures of porcine alveolar macrophages (PAM). Farm production recovered after eight weeks of losses. In order to evaluate the persistence of infection in the herd, five SPF-piglets were introduced in two different sections of the PRRS-affected barn four months after the disappearance of clinical symptoms, and two others were placed in a neighboring building with apparently healthy farrow-to-finnish pigs. Clinical signs, body temperature, humoral immune response, virological and histopathological findings were recorded over a 42-day period. Clinical signs were evident in all of the sentinels and prolonged fever (> or = 40 degrees C) was recorded one day post-exposure (PE). Antibody titers to PRRS virus could be detected by IIF on PAM seven days PE, and reached 1:1024 by day 21 PE. Three of the sentinels developed significant virus neutralizing antibody titers (> 1:8 to < or = 1:128) by day 35 PE. In all cases, the virus could be isolated from the serum between day 7 and 42 PE. Thus, the virus and specific antibodies coexisted for several weeks. Lesions of interstitial pneumonia was demonstrated in few animals. In experimental inoculation studies, the viral strain isolated from the sentinel pigs produced severe reproductive disorders in two sows inoculated at 95 days of gestation.(ABSTRACT TRUNCATED AT 250 WORDS)     

猪繁殖与呼吸综合征病毒接触感染仔猪模型的建立

为了建立猪繁殖与呼吸综合征病毒(PRRSV)接触感染模型,将易感仔猪与PRRSV人工感染仔猪在隔离器中同居饲养,每日观察记录体温变化和临床症状,定期采集血清和粪便样品,用荧光定量RT-PCR检测病毒血症和粪便排毒水平;对病死猪和试验结束迫杀猪进行剖检和肺脏组织病理学检查,并检测各器官病毒载量和组织分布。结果显示:接触感染仔猪出现高热和PRRS症状时间分别较人工接种猪推迟2~3d;第3天起血清检测PRRSV阳性,第10天病毒血症上升至较高水平(108.6拷贝/mL);第5天起粪便检测PRRSV阳性,第7天粪便排毒量上升至较高水平(103.9拷贝/0.1g);3头接触感染仔猪分别于同居后第12天和第13天死亡,较人工接种猪推迟1~2d,病死猪的主要器官病毒载量以及剖检和肺脏显微病理变化与人工接种病死猪相似。这些结果表明,本研究建立的接触感染仔猪模型可用于PRRS疫苗免疫效果评价。     

Pathogenesis of porcine reproductive and respiratory syndrome virus infection in mid-gestation sows and fetuses.

Two experiments were undertaken to evaluate whether porcine reproductive and respiratory syndrome (PRRS) virus was able to cross the placenta and infect midgestation fetuses following intranasal inoculation of sows and whether PRRS virus directly infected fetuses following in utero inoculation. In experiment 1, eight sows between 45 and 50 days of gestation were intranasally inoculated with PRRS virus (ATCC VR-2332), and four control sows were inoculated with uninfected cell culture lysate. Virus inoculated sows were viremic on postinoculation (PI) days 1, 3, 5, 7 and 9, shed virus in their feces and nasal secretions, and became leukopenic. Sixty-nine of 71 fetuses from principal sows euthanized on PI day 7, 14 or 21 were alive at necropsy and no virus was isolated from any of the fetuses. Two principal sows that farrowed 65 and 67 days PI delivered 25 live piglets and three stillborn fetuses. The PRRS virus was isolated from two live piglets in one litter. In experiment 2, laparotomies were performed on five sows between 40 and 45 days of gestation and fetuses were inoculated in utero with either PRRS virus alone, PRRS virus plus a swine serum containing PRRS antibodies, or uninfected cell culture lysate. Three sows were euthanized on PI day 4 and two sows on PI day 11. Viral replication occurred in fetuses inoculated with virus alone and was enhanced in fetuses inoculated with virus plus antibody. No virus was isolated from control fetuses.(ABSTRACT TRUNCATED AT 250 WORDS)     

CD163片段在PRRSV感染过程中的作用

使用Scanprosite软件对猪肺泡巨噬细胞的CD163蛋白结构和功能域进行预测,并对其进行分段克隆,构建原核表达载体pET-28a(Y-P1,Y-P2,Y-P3),片段位置Y-P1(160~798bp),Y-P2(790~20 146bp),Y-P3(2 143~3 084bp)。利用镍柱纯化重组蛋白,免疫小鼠获得特异性抗体。通过病毒阻断试验,验证Y-P1,Y-P2,Y-P3的PRRSV阻断情况;构建真核表达载体pCD163,P1(1~798bp),P2(790~2 046bp),P3(2 023~3 345bp)分别转染PRRSV非易感细胞系BHK-21,验证其PRRSV感染情况。结果显示,经PCR、双酶切及测序鉴定构建的原核和真核表达载体是正确的,SDS-PAGE检验Y-P1,Y-P2,Y-P3蛋白大小分别为27 000,46 000,39 000。Y-P2蛋白,抗Y-P2的小鼠血清能够阻断PRRSV感染Marc-145细胞;转染pCD163的细胞系能够感染PRRSV,而P1,P2,P3片段不感染PRRSV。CD163其790~2 046bp可能是PRRSV感染细胞与CD163受体作用位点。     

猪繁殖与呼吸综合征病毒感染的诊断及其分子特点

采用流行病学调查、临床症状观察、病理学检查、病毒分离、分离毒株测序以及遗传演化分析的方法,对北京及其周边地区4个猪场剖检的4头猪进行分析。结果:流行病学和临床症状表现为急性高热性传染病;病理学观察为非化脓性脑炎和间质性肺炎等多器官严重的病理变化;RT-PCR和病毒分离确定该疫情的主要病原是猪繁殖与呼吸综合征病毒(PRRSV)。与典型PRRSV感染不同的是:成年猪感染率达50%以上,病死率达80%以上;PRRSV分离株全基因序列分析表明属于PRRSV北美洲型,特别是PRRSV NSP2不连续缺失30个氨基酸,说明此次流行的PRRSV毒株可能为高致病性毒株。     

Effect of porcine reproductive and respiratory syndrome virus infection on the clearance of Haemophilus parasuis by porcine alveolar macrophages.

Porcine reproductive and respiratory syndrome virus (PRRSV) infection in young piglets is frequently associated with secondary infection due to various pathogens, especially those of the respiratory tract. One of the most important mechanisms in respiratory diseases is related to the alteration of function of porcine alveolar macrophages (PAMs). The objective of this study was to determine how PRRS virus infection affects the capabilities of PAMs in the phagocytosis and destruction of Haemophilus parasuis. Phagocytosis percentages were determined in vitro and ex vivo, after collected PAMs were directly exposed to the virus of if PAMs were collected from piglets previously infected with PRRSV. In vitro experiments demonstrated that H. parasuis uptake by PAMs is only increased in the early stages of PRRSV infection (2 h post-infection). In contrast, in the ex vivo experiments it was shown that PAMs from PRRSV-infected piglets do not seem to change in their phagocytic rate until the later stages of infection. Together with a decrease in the phagocytic rate, a marked decrease in the functional ability of PAMs to kill bacteria was observed 7 d post-infection. It is hypothesized that when animals are exposed to PRRSV, there is a marked decrease in the functional ability of PAMs to kill bacteria through the release of superoxide anion, indicating a possible negative effect of the virus, at least at the macrophage level.     

An assessment of sanitation protocols for commercial transport vehicles contaminated with porcine reproductive and respiratory syndrome virus

The objective of this study was to develop and test a rapid (< 2 h) sanitation protocol designed for porcine reproductive and respiratory syndrome virus (PRRSV) positive commercial transport vehicles involving cold water washing and disinfection via fumigation using scale models of weaned pig trailers. The study consisted of 2 phases. Following experimental contamination of model trailers with PRRSV MN 30-100 (5 x 10(5)TCID50), phase 1 evaluated the presence or absence of PRRSV RNA by polymerase chain reaction (PCR) on swabs collected from the trailer interiors 0, 60, and 90 min after treatment. Phase 2 consisted of evaluating the infectivity of trailers 90 min posttreatment by monitoring changes in the PRRSV-status of naive sentinel pigs housed for 2 h. Treatments included washing only (treatment 1), washing plus formaldehyde fumigation (treatment 2), washing plus fumigation with glutaraldehyde-quaternary ammonium chloride (treatment 3), and washing plus overnight drying (treatment 4). Porcine reproductive and respiratory syndrome virus RNA was detected in all trailers (20 out of 20 replicates) at 60 and 90 min following the application of treatments 1 and 2. These trailers also contained infectious PRRSV, as determined by the infection of naive pigs housed in treated trailers and the testing of organic debris collected from the interior of trailers by swine bioassay. At 90 min posttreatment, all trailers treated with glutaraldehyde-quaternary ammonium chloride were PCR-negative, non-infectious to sentinel pigs, and swine bioassay negative. Similar results were observed in trailers allowed to dry for 8 h. Under the conditions of this study, it appears certain disinfectants may possess different levels of efficacy against PRRSV and PRRSV-positive models may be effectively sanitized in the absence of overnight drying.     

广西猪繁殖与呼吸综合征病毒感染状况调查

采用RT-PCR技术,对2004年1月至2005年4月期间,广西13个市104个疑似猪繁殖与呼吸综合征病毒(PRRSV)感染猪场,无菌采取231头病、死猪的组织病料(肺脏、淋巴结、脾脏)进行了病毒检测。同时,对鉴定为PRRSV阳性的组织病料和猪场进行了猪圆环病毒2型(PCV2)、猪瘟病毒(CSFV)和猪伪狂犬病毒(PRV)的检测,以确定猪群中PRRSV与PCV2、CSFV和PRV混合感染情况。结果从12个市的115份组织病料中检出PRRSV,病料的平均阳性率为49.78%(115/231),猪场的平均阳性率为61.54%(64/104),不同地市有一定的差异。PRRSV与PCV2、CSFV和/或PRV二重或多重混和感染的组织病料总数为53份,猪场总数为39个,混合感染的组织病料和猪场的总阳性率分别为22.94%(53/231)和37.50%(39/104)。混合感染的组织病料占PRRSV阳性组织病料的46.09%(53/115),混合感染的猪场占PRRSV阳性猪场的60.94%(39/64)。其中以PCV2和PRRSV混合感染的组织病料和猪场数最多。由此可见,PRRSV感染在广西猪场已普遍存在,与其他病毒混合感染现象逐渐趋向复杂化。     

Pathogenesis and prevention of placental and transplacental porcine reproductive and respiratory syndrome virus infection

Porcine reproductive and respiratory syndrome virus (PRRSV)-induced reproductive problems are characterized by embryonic death, late-term abortions, early farrowing and increase in number of dead and mummified fetuses, and weak-born piglets. The virus recovery from fetal tissues illustrates transplacental infection, but despite many studies on the subject, the means by which PRRSV spreads from mother to fetus and the exact pathophysiological basis of the virus-induced reproductive failure remain unexplained. Recent findings from our group indicate that the endometrium and placenta are involved in the PRRSV passage from mother to fetus and that virus replication in the endometrial/placental tissues can be the actual reason for fetal death. The main purpose of this review is to clarify the role that PRRSV replication and PRRSV-induced changes in the endometrium/placenta play in the pathogenesis of PRRSV-induced reproductive failure in pregnant sows. In addition, strategies to control placental and transplacental PRRSV infection are discussed.     

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…     

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.     

Envision免疫组化技术在诊断猪繁殖与呼吸综合征中的应用

为探讨免疫组织化学Envision法在诊断猪繁殖与呼吸综合征(PRRS)中应用的可行性,采用免疫组织化学Envision二步法对30例疑似PRRS病猪肺组织的病毒抗原进行定位、半定量检测。结果表明,Envision法可原位检测病猪肺组织PRRSV抗原的分布;其抗原的阳性表达主要出现在肺巨噬细胞胞浆内,其次为肺泡上皮细胞和细支气管黏膜上皮细胞。依据阳性细胞百分率,进行染色结果判断,检出阳性病猪23例,阳性检出率为76.7%。试验证明,Envision法具有高敏感、低背景、快速简便的特点,可在PRRS诊断中推广应用。     

猪繁殖与呼吸综合征病毒和致病性沙门氏菌的混合感染

2003年9～11月某猪场发生母猪持续性繁殖障碍和7日龄仔猪呼吸困难、寒颤、下痢、体温升高和实质性器官出血为主要特征的疾病。3头发病仔猪的病变组织用RT-PCR检测。均为猪繁殖与呼吸综合征病毒(PRRSV)阳性；由3头发病猪病料中所分离的细菌。可于普通琼脂、伊红美蓝和麦康凯琼脂培养基上生长，革兰氏染色阴性，形态与组织触片镜检及生化反应特性与沙门氏菌一致，可致死小鼠。仅对头孢唑啉呈中度敏感。结合该病的流行病学调查、临床症状、病理剖检，确诊为猪繁殖与呼吸综合征病毒(PRRSV)和致病性沙门氏菌混合感染。