2004～2006年加拿大安大略省和魁北克省猪出现与严重猪圆环病毒相关疾病有关的猪圆环病毒2型新毒株

2004年晚秋,加拿大安大略省保育和生长育肥猪发生了比通常猪圆环病毒2型（PCV-2）感染更严重的PCV-2相关疾病（PCVAD）。病变空前严重,包括弥散性支气管间质性肺炎、肉芽肿性肠炎、脉管炎、间质性肾炎和新病变脾梗塞。一些感染猪群的死亡率高达50%。此病与突然出现的聚合酶链反应（PCR）限制性片段长度多态性（RFLP）321型PCV-2变种有关。开放阅读框（ORF2）和全基因组序列的种系发生比较,显示为与以前占优势的RFLP422型病毒不同的新变种,与曾发生在法国和其他欧洲国家及亚洲国家的相似。随后的回顾性研究,与较早的RFLP422型毒株相比,猪自然感染RFLP321型的淋巴结、脾、肺、小肠和肾的组织学病变发生率有统计学的明显增加。基于淋巴结免疫组织化学（IHC）染色,猪感染RFLP321型毒株,与较早的RFLP422型毒株相比,病毒载负量也显示统计学的明显增加。RFLP321型毒株感染猪后的毒力增强,可能与新PCV-2毒株的遗传差异有关,此病毒目前为安大略和魁北克省猪占优势的PCV-2毒株。     

The clinical expression and emergence of porcine circovirus 2

Since its discovery and characterization in western Canada in 1995, the distribution of post-weaning multisystemic wasting syndrome (PMWS) has grown and the syndrome is undoubtedly recognized worldwide. More recently, there is a heightened interest in several potentially related conditions including porcine dermatitis and nephropathy syndrome (PDNS), proliferative and necrotizing pneumonia (PNP), congenital tremors (CT-AII), perinatal myocarditis and reproductive failure. Although the causality of porcine circovirus type 2 (PCV2) has recently been proven [Vet. Pathol. 37 (2000) 254; J. Comp. Pathol. 122 (2000) 9; J. Vet. Diagn. Invest. 13 (2001) 185], PCV2 is present in both diseased and healthy pig populations. Furthermore, recent serologic studies have shown that there are no significant differences in PCV2 specific IgG titres between PMWS clinical and non-clinical herds (Harding et al., in press). Clearly, the transmission, epidemiology, and factors triggering clinical expression are poorly understood. With these uncertainties in mind, the objective of this paper is to review the clinical expression and emergence of PMWS and the potentially associated disease syndromes including PDNS.     

猪圆环病毒病Ⅱ型的诊断与防治

某地村民宋某自养商品猪126头,体重在30～40kg。从2004年7月10日开始陆续发病,其主要特征为猪的体质下降,消瘦,体温升高,出血、呼吸困难,贫血,个别猪出现咳嗽,腹泻,黄疸等,临床用药效果甚微。我们依据流行病学、临床表现、病理变化以及实验室诊断,综合诊断为猪圆环病毒病。1发病情况该批猪是5月份从外地购入,一周后分别进行了猪瘟、仔猪副伤寒、猪肺疫、猪丹毒、猪水肿病以及口蹄疫病的免疫接种。到7月10日前没有发生任何疫病,吃食、精神均正常。7月10日后陆续发病,到7月22日发病36头,死亡7头,在饲养环节中未发现异常。2临床表现病初体温升…     

The emergence of porcine circovirus 2b genotype (PCV-2b) in swine in Canada

Since late 2004, the swine industry in the province of Quebec has experienced a significant increase in death rate related to postweaning multisystemic wasting syndrome (PMWS). To explain this phenomenon, 2 hypotheses were formulated: 1) the presence of a 2nd pathogen could be exacerbating the porcine circovirus 2 (PCV-2) infection, or 2) a new and more virulent PCV-2 strain could be infecting swine. In 2005, 13 PMWS cases were submitted to the Quebec provincial diagnostic laboratory and PCV-2 was the only virus that could be found consistently by PCR in all 13 samples. The PCR detection results obtained for other viruses revealed the following: 61.5% were positive for porcine reproductive and respiratory syndrome virus, 30.8% for swine influenza virus, 15.4% for porcine parvovirus, 69.2% for swine torque teno virus (swTTV), 38.5% for swine hepatitis E virus (swHEV) and 84.6% for Mycoplasma hyorhinis; transmissible gastroenteritis virus and porcine respiratory coronavirus (TGEV/PRCV) was not detected. Sequences of the entire genome revealed that these PCV-2 strains belonged to a genotype (named PCV-2b) that has never been reported in Canada. Further sequence analyses on 83 other Canadian PCV-2 positive cases submitted to the provincial diagnostic laboratory during years 2005 and 2006 showed that 79.5% of the viral sequences obtained clustered in the PCV-2b genotype. The appearance of the PCV-2b genotype in Canada may explain the death rate increase related to PMWS, but this relationship has to be confirmed.     

The effect of vaccination against Porcine reproductive and respiratory syndrome virus (PRRSV) on the Porcine circovirus-2 (PCV-2) load in porcine circovirus associated disease (PCVAD) affected pigs

A diagnostic project was initiated across the United States in 2006 to improve the understanding of porcine circovirus associated diseases (PCVAD) as well as to identify co-factors in PCVAD-affected farms. A Porcine circovirus-2 (PCV-2) DNA real-time polymerase chain reaction quantitation (qPCR) was established according to a published method and sera from a total of 23pig farms across the United States were examined for viral loads for PCV-2 and analyzed for any possible effects of Porcine reproductive and respiratory syndrome virus (PRRSV) vaccination on this parameter. Vaccination against PRRS resulted in significantly lower viral loads for PCV-2 in animals 13 wk or older compared with nonvaccinated animals, but vaccination of pigs against PRRS had no effect on qPCR results for PCV-2 in 4- to 12-week-old pigs. Interestingly, PRRS vaccinates had significantly lower viral loads when peak wasting disease was observed in the herds. The qPCR method for PCV-2 proved to be an important tool for help in the antemortem diagnosis of PCVAD as well as in the monitoring of infection dynamics.     

Case-control study on the association of porcine circovirus type 2 and other swine viral pathogens with postweaning multisystemic wasting syndrome.

A field-based case-control study was conducted to assess the strength of association of porcine circovirus type 2 (PCV2) and some major swine viruses with postweaning multisystemic wasting syndrome (PMWS). Cases were defined as individual pigs with a clinical history of progressive weight loss and histopathological lesions characteristic of PMWS. Controls were pigs without clinical signs and histopathological lesions typical of PMWS. A total of 31 cases and 56 controls was identified from diagnostic submissions. Serum and various tissues were collected from all animals and assayed for PCV, porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus, porcine enterovirus types 1-3, swine influenza virus, porcine respiratory coronavirus, transmissible gastroenteritis virus, porcine endogenous retrovirus, porcine lymphotropic herpesvirus type 1, and bovine viral diarrhea virus. The proportion of case and control pigs positive for each virus was determined and statistically compared for determining the strength of the association that each virus had with PMWS individually or in combinations. Porcine circovirus type 2 had the strongest association (OR = 9.3, P = 0.006) with PMWS among the viruses tested for. Risk for PWMS was much higher (OR = 31.2, P = 0.0009) if the animal was concurrently infected with PCV2 and PRRSV, suggesting that development of PMWS may be enhanced by cofactor(s). Because PCV2 was also found in 62.5% of the controls, PCV2 from 5 cases and 4 controls were selected and genetically compared. No significant genetic difference was observed between PCV2 from PMWS and control pigs.     

一例猪瘟与猪圆环病毒2型混合感染的诊治

在生猪养殖过程中,多种病原混合感染的情况较为常见,并这种混合感染也导致传统的临床诊断容易造成误诊或漏诊,继而延误诊断治疗从而造成经济损失.2020年8月初,孟连县某养猪场保育猪群零散发病,临床主要表现为发热、咳喘、打堆、被毛凌乱无光泽、局部皮肤发绀、消瘦、饮食欲下降、生长滞缓,个别猪只伴便秘和腹泻交替.病理剖检变化显示...     

重组新城疫病毒表达猪圆环病毒2型Cap蛋白的研究

为构建表达猪圆环病毒2型(PCV2)衣壳蛋白(Cap)的重组新城疫病毒(NDV),本研究利用NDV LaSota弱毒疫苗株为活病毒载体,通过反向遗传操作系统构建出表达PCV2 Cap的重组病毒(rLa-PCV2/Cap),以及表达删除核定位信号(NLS)的Cap(Cap△41)的重组病毒(rLa-PCV2/Cap△41),并对外源蛋白的表达效果进行了检测.通过间接免疫荧光试验证明,两种重组病毒感染的BHK-21细胞中,PCV2 Cap蛋白以及删除NLS的Cap△41蛋白均获得正确表达;其中,完整的Cap蛋白定位于核内,而删除NLS的Cap△41蛋白则定位于胞浆中,本研究结果为研究新型PCV2疫苗奠定基础.     

Experimental reproduction of severe disease in CD/CD pigs concurrently infected with type 2 porcine circovirus and porcine reproductive and respiratory syndrome virus

Three-week-old cesarean-derived colostrum-deprived (CD/CD) pigs were inoculated with porcine circovirus type 2 (PCV2, n = 19), porcine reproductive and respiratory syndrome virus (PRRSV, n = 13), concurrent PCV2 and PRRSV (PCV2/PRRSV, n = 17), or a sham inoculum (n = 12) to compare the independent and combined effects of these agents. Necropsies were performed at 7, 10, 14, 21, 35, and 49 days postinoculation (dpi) or when pigs became moribund. By 10 dpi, PCV2/PRRSV-inoculated pigs had severe dyspnea, lethargy, and occasional icterus; after 10 dpi, mortality in this group was 10/11 (91%), and all PCV2/ PRRSV-inoculated pigs were dead by 20 dpi. PCV2-inoculated pigs developed lethargy and sporadic icterus, and 8/19 (42%) developed exudative epidermitis; mortality was 5/19 (26%). PRRSV-inoculated pigs developed dyspnea and mild lethargy that resolved by 28 dpi. Microscopic lesions consistent with postweaning multisystemic wasting syndrome (PMWS) were present in both PCV2- and PCV2/PRRSV-inoculated pigs and included lymphoid depletion, necrotizing hepatitis, mild necrotizing bronchiolitis, and infiltrates of macrophages that occasionally contained basophilic intracytoplasmic inclusion bodies in lymphoid and other tissues. PCV2/ PRRSV-inoculated pigs also had severe proliferative interstitial pneumonia and more consistent hepatic lesions. The most severe lesions contained the greatest number of PCV2 antigen-containing cells. PRRSV-inoculated pigs had moderate proliferative interstitial pneumonia but did not develop bronchiolar or hepatic lesions or lymphoid depletion. All groups remained seronegative to porcine parvovirus. The results indicate that 1) PCV2 coinfection increases the severity of PRRSV-induced interstitial pneumonia in CD/CD pigs and 2) PCV2 but not PRRSV induces the lymphoid depletion, granulomatous inflammation, and necrotizing hepatitis characteristic of PMWS.     

Detection of porcine reproductive and respiratory syndrome virus, porcine circovirus type 2, swine influenza virus and Aujeszky's disease virus in cases of porcine proliferative and necrotizing pneumonia (PNP) in Spain

Proliferative and necrotizing pneumonia (PNP) is a severe form of interstitial pneumonia characterised by hypertrophy and proliferation of pneumocytes type 2 and presence of necrotic cells within alveoli lumen. Many viral agents have been linked to PNP aetiology, with especial emphasis on porcine reproductive and respiratory syndrome virus (PRRSV). To gain knowledge on PNP causality, a retrospective study on 74 PNP cases from postweaning pigs from Spain was carried out. Coupled with histopathological examinations, the presence of porcine circovirus type 2 (PCV2) by in situ hybridization (ISH), and PRRSV, swine influenza virus (SIV) and Aujeszky's disease virus (ADV) by immunohistochemical (IHC) methods, were investigated. PCV2 was the most prevalent viral agent in PNP cases (85.1%) followed by PRRSV (44.6%); 39.1% of PNP cases showed PCV2 as the solely detected agent, while only 4.1% had PRRSV as the unique pathogen. SIV and ADV were very sporadically detected in PNP cases, and always in co-infection with PCV2. Therefore, present data indicate that PCV2 is the most important aetiological agent in PNP cases from Spain and that PRRSV is not essential for the development of PNP. Taking into account the presented results and available literature, we suggest that PCV2 is possibly the main contributor to PNP cases in Europe while PRRSV could play a similar role in North America.     

One dose of a porcine circovirus 2 subunit vaccine induces humoral and cell-mediated immunity and protects against porcine circovirus-associated disease under field conditions

This study investigated the efficacy of a one-dose porcine circovirus 2 (PCV2) subunit vaccine based on the PCV2 Cap protein expressed in a baculovirus system on two different farms at which a history of porcine circovirus-associated disease (PCVD) was present. Morbidity, mortality, average daily weight gain, carcass weight, PCV2 load in serum and vaccine immunogenicity were assessed. Serology to porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae was performed. A double-blind, randomised, and controlled field trial was performed distributing 818 piglets between two treatment groups. At inclusion (weaning at 21 ± 3 days of age), 408 animals (group B) received a 2-mL intramuscular dose of Porcilis PCV(?) (vaccinated group). Controls (group A, 410 pigs) received 2 mL of the adjuvant Diluvac Forte(?) intramuscularly. Weights were recorded at inclusion and at 12 and 26 weeks of age, and the average daily weight gain (ADWG) was calculated. The carcass weights of the pigs from farm 2 were recorded at slaughter (274 days old). All dead animals (died or culled) underwent autopsy to classify them as PMWS-affected or not. At each farm, blood samples were taken from 22 pigs/group for serologic studies. A beneficial effect was found after vaccination with a single dose of a PCV2 Cap vaccine against PCVD. The vaccination reduced the mortality rate and morbidity, reduced PCV2 viremia and viral load, improved productive performances (e.g. ADWG: +70 g/day between 12 and 26 weeks of age when viremia and the specific disease occurred) as well as carcass weight at slaughter age (+4.5 kg). These effects were associated with virologic and clinical protection from the immunogenicity of the vaccine measured as activation of both a humoral and a cellular immune response.     

The association between submission counts to a veterinary diagnostic laboratory and the economic and disease challenges of the Ontario swine industry from 1998 to 2009

An intuitive assumption is to believe that the number of submissions made to a veterinary diagnostic laboratory is dictated by the financial state of the industries using the laboratory. However, no research is available to document how the economics of a food animal industry affects laboratory submissions and therefore disease monitoring and surveillance efforts. The objective of this study was to determine if economic indices associated with the Ontario swine industry can account for the variability seen in these submissions. Retrospective swine submissions made to the Animal Health Laboratory at the University of Guelph, Guelph, Ontario from January 1998 to July 2009 were compiled. The following economic, demographic, and health variables impacting Ontario swine production were selected for analysis: auction price, lean-hog futures, currency exchange rate, price of corn, an outbreak of porcine circovirus type-2 associated diseases (PCVAD), government incentive program, number of farms in province, and average farm size. All independent variables identified by unconditional associations to have a significance of P≤0.2 with the outcome of monthly submission count were included in a multivariable negative binomial model. A final model was identified by a backwards elimination procedure. A total of 30,432 swine submissions were recorded. The mean frequency of monthly submissions over 139months was 212.9 (SD=56.0). After controlling for farm size, the number of pigs in Ontario, higher submission counts were associated with a weaker CAD$ versus US$, higher auction prices, and a PCVAD outbreak (P<0.001). The results suggest that both economic volatility and disease outbreaks in the Ontario swine industry drive submissions to the laboratory. In conclusion, lab submissions are a useful source of animal health data for disease surveillance; however, surveillance activities should also monitor the economics of the industry.     

Mycoplasma hyopneumoniae bacterins and porcine circovirus type 2 (PCV2) infection: induction of postweaning multisystemic wasting syndrome (PMWS) in the gnotobiotic swine model of PCV2-associated disease

Groups (5 to 15 per group) of gnotobiotic swine were infected oronasally with porcine circovirus type 2 (PCV2) at 3 days of age and then given 1 of 6 different commercial Mycoplasma hyopneumoniae (M. hyopneumoniae) bacterins as either a single dose (7 d of age, 1 application products) or 2 doses (7 and 21 d of age, 2 application product). Control groups received PCV2 alone (n = 9) or were infected with PCV2 and immunized twice with keyhole limpet hemocyanin (KLH) emulsified in incomplete Freund's adjuvant (ICFA) (n = 7). Five of 7 (71%) PCV2-infected piglets immunized with KLH/ICFA developed mild or overt PMWS, whereas none of 9 piglets infected with PCV2 alone developed PMWS. Five of 12 (42%) piglets vaccinated with a commercial bacterin containing mineral oil adjuvant developed PMWS following vaccination. None of the PCV2-infected piglets in the other bacterin-vaccinated groups developed PMWS in this model of PCV2-associated disease. This difference in prevalence of PMWS in piglets given the mineral oil-adjuvanted M. hyopneumoniae bacterin and the other M. hyopneumoniae bacterin vaccination groups was statistically significant (P < 0.05).     

A live-attenuated and an inactivated chimeric porcine circovirus (PCV)1-2 vaccine are both effective at inducing a humoral immune response and reducing PCV2 viremia and intrauterine infection in female swine of breeding age

The objective of this pilot study was to determine the efficacy of inactivated (1 or 2 dose) and live-attenuated chimeric porcine circovirus (PCV)1-2 vaccines in sows using the PCV2-spiked semen model. Thirty-five sows were randomly divided into 6 groups: negative and positive controls, 1 dose inactivated PCV1-2 vaccine challenged (1-VAC-PCV2), 2 dose inactivated PCV1-2 vaccine challenged (2-VAC-PCV2), 1 dose live-attenuated PCV1-2 vaccine unchallenged (1-LIVE-VAC), and 1 dose live-attenuated PCV1-2 vaccine challenged (1-LIVE-VAC-PCV2). The inactivated PCV1-2 vaccine induced higher levels of PCV2-specific antibodies in dams. All vaccination strategies provided good protection against PCV2 viremia in dams, whereas the majority of the unvaccinated sows were viremic. Four of the 35 dams became pregnant: a negative control, a positive control, a 2-VAC-PCV2 sow, and a 1-LIVE-VAC-PCV2 sow. The PCV2 DNA was detected in 100%, 67%, and 29% of the fetuses obtained from the positive control, inactivated vaccinated, or live-attenuated vaccinated dams, respectively. The PCV2 antigen in hearts was only detectable in the positive control litter (23% of the fetuses). The PCV1-2 DNA was detected in 29% of the fetuses in the litter from the 1-LIVE-VAC-PCV2 dam. Under the conditions of this pilot study, both vaccines protected against PCV2 viremia in breeding age animals; however, vertical transmission was not prevented.     

Retrospective Study: Incidence of ionized hypocalcemia in septic dogs and its association with morbidity and mortality: 58 cases (2006–2007)

Objective – To determine the incidence rate and prognostic significance of ionized hypocalcemia (iHCa) among septic dogs. Design – Retrospective study Setting – Veterinary teaching hospital Animals – Fifty‐eight septic dogs that were presented to Cornell University Hospital for Animals between January 2006 and December 2007. Procedure – Cases were diagnosed with sepsis if they exhibited 2 or more criteria of the systemic inflammatory response syndrome with a concurrent documented infectious focus. Cases were excluded if diagnosed with a concurrent illness reportedly associated with calcium derangements. Lowest, mean, and highest blood ionized calcium concentrations were recorded and statistically analyzed for an association with morbidity, as measured by duration of hospitalization and number of blood product transfusions, and outcome. In addition, the incidence rate of iHCa was recorded. Results – Of the 58 cases included in this study, iHCa was documented in 4 of 6 (67%) patients that died, 5 of 19 (26%) euthanized patients and 5 of 33 (15%) patients that survived to discharge, with an overall incidence of 24%. Dogs that died during hospitalization had more severe iHCa than patients that were discharged or euthanized as well as significantly lower mean ionized calcium concentrations than patients who were discharged. Severity of iHCa was also associated with a longer duration of hospitalization. The highest ionized calcium concentration was not associated with outcome. Conclusion and Clinical Relevance – This study is the first to document the incidence of iHCa among septic dogs. Because both low mean ionized calcium and the lowest documented ionized calcium concentration are associated with poor outcome, it is likely that both the severity and duration of hypocalcemia are important in these patients. Further prospective studies investigating the prognostic significance, etiology and treatment of iHCa among septic veterinary patients are needed to better understand its role in sepsis.