An ORF2 protein-based ELISA for porcine circovirus type 2 antibodies in post-weaning multisystemic wasting syndrome

Porcine circovirus type 2 (PCV2) plays a crucial role in the pathogenesis of post-weaning multisystemic wasting syndrome (PMWS) in swine. As PCV2 displays significant homology with PCV1 (a non-pathogenic virus) at the nucleotide and amino-acid level, a discriminative antigen is needed for specific serological diagnosis. The ORF2-encoded capsid protein from PCV2 was used to develop an indirect enzyme-linked immunosorbent assay (ELISA). GST-fused capsid protein from PCV2 and GST alone (both expressed in recombinant baculovirus-infected cells) were used as antigens for serodiagnosis. The specificity of the ELISA for detection of PCV2 antibodies was demonstrated in sera from pigs experimentally infected with PCV1, PCV2 and other swine viruses. The semi-quantitative nature of the test was evaluated versus an immunoperoxidase monolayer assay (IPMA). The ELISA was performed on 322 sera from pigs in eight Brittany herds and compared with IPMA. The sensitivity (98.2%) and specificity (94.5%) of this test were considered suitable for individual serological detection. High PCV2 seroprevalence was found in sows and pigs at the end of the growth phase (18-19 weeks) in all eight herds. The seroprevalence in piglets (11-17 weeks) was statistically correlated with clinical symptoms of PMWS (93% in affected versus 54%, in non-affected farms). A cohort study performed in PMWS-free farms showed that 57% of piglets exhibited active seroconversion after 13 weeks, indicating that PCV2 infection occurred earlier in PMWS-affected piglets.     

规模猪场猪瘟野毒感染情况调查

利用ELISA方法对广西6个规模猪场送检的1626份猪血清进行猪瘟野毒感染情况调查.结果6个规模猪场均存在猪瘟野毒感染,其中母猪感染率为7.86%～29.21%,平均为17.52%,种公猪感染率为0%～23.52%,平均为11.83%,育肥猪感染率为5%～22.45%,平均为15.5%,断奶仔猪感染率为8.24%～18.57%,平均为12.69%.此检测结果与猪场临床发病情况基本一致,病猪多表现为繁殖障碍型、温和型的非典型猪瘟.     

The epidemiology of Leptospira interrogans serovar pomona in grower pig herds

An epidemiological study on four farms showed that leptospiral infection became apparent in piglets from 12 weeks of age. The intensity of leptospiruria was greatest in the first 3-4 weeks of infection and from then on declined and became intermittent. Factors affecting the cultural and serological prevalence of infection of the piglets were found to be the standard of hygiene and variations in the titre of the dam. Dams with high titres gave better protection to their young than did those with low titres. The spread of infection within piggeries was encouraged by mixing infected with uninfected pigs, which resulted in epidemics within the pens. Transmission from infected to susceptible grower pigs continuously occurred in grower houses, with a constant proportion of pigs becoming infected each week.     

Profiles of seroconversion to porcine circovirus type 2 in herds affected and not affected by postweaning multisystemic wasting syndrome

The aim of the present study was to explore the usefulness of serological methods in the diagnosis of postweaning multisystemic wasting syndrome (PMWS). The study was carried out in 4 PMWS-affected and 6 control farms. Based on the serological profiles, infection with porcine circovirus type 2 (PCV2) was determined to take place at 3-7 weeks of age in the PMWS-affected and at 3-11 weeks of age in the control farms. To compare the dynamics of seroconversion to PCV2 among farms, cross-sectional serological profiles were normalised in relation to the inferred age of infection. The results indicated that the proportion of seropositive pigs increased significantly slower in the affected herds. The most pronounced difference was observed about 4 weeks after infection, when the proportion of seropositive pigs ranged from 0 to 53.3% and from 70 to 100% in PMWS-affected and control herds, respectively. Mean antibody titres at that age were also significantly lower in the affected farms. These observations suggest a delay in the production of PCV2-specific antibodies and indicate that serological methods may be helpful in identifying herds with a high risk of PMWS.     

模猪场猪瘟野毒感染情况调查

利用ELISA方法对广西6个规模猪场送检的1626份猪血清进行猪瘟野毒感染情况调查。结果6个规模猪场均存在猪瘟野毒感染,其中母猪感染率为7.86%~29.21%,平均为17.52%,种公猪感染率为0%~23.52%,平均为11.83%,育肥猪感染率为5%~22.45%,平均为15.5%,断奶仔猪感染率为8.24%~18.57%,平均为12.69%。此检测结果与猪场临床发病情况基本一致,病猪多表现为繁殖障碍型、温和型的非典型猪瘟。     

Risk factors for Salmonella seroconversion of fattening pigs in farrow-to-finish herds

We did a prospective observational 9-month long study to quantify risk factors of managerial and hygiene practices, and pig-health status for Salmonella seroconversion of fattening pigs reared in subclinically infected French farrow-to-finish farms. During the fattening phase, 2,649 pigs belonging to the same batch of contemporary pigs, from 89 conventional farrow-to-finish farms were individually followed and regularly blood sampled on a monthly basis. Farm recruitment was based on the farmer's willingness to cooperate. Pig status was assessed using an indirect ELISA test. Evolution of the serological status was studied by means of survival analysis. A Cox proportional-hazards model, taking into account the clustering of animals at the farm level, was used to examine the effects of explanatory variables on the time to Salmonella seroconversion of pigs. Applying group level antibiotic treatment to the pigs during the fattening period (Hazard Ratio (HR) = 2.4; 95% CI: 1.7, 3.4) was identified as a risk factor for Salmonella seroconversion, as the presence of residual Salmonella contamination in the fattening pen before placing the pigs into the pens (HR = 1.9; 95% CI: 1.2, 2.9). Porcine reproductive and respiratory syndrome virus (PRRSV) seropositivity during the fattening period also indicated an increased hazard for seroconversion (HR = 1.6; 95% CI: 1.1, 2.5). The batch size was identified as a risk factor for Salmonella seroconversion: the higher the number of pigs was in the fattening room followed, the higher was the risk (HR(+10 pigs) = 1.05 for a 10-pig increment; 95% CI: 1.03, 1.06). The biosecurity measures of wearing specific clothes before entering the facilities (HR = 0.5; 95% CI: 0.3, 0.9) and enclosing the pig farm facilities were protective (HR = 0.4; 95% CI: 0.2, 0.8).     

黑龙江省东部地区猪蓝耳病流行检测分析

为了解黑龙江省东部地区猪蓝耳病(PRRS)的流行情况,从黑龙江省东部地区的牡丹江市、宁安市、海林市、绥芬河市、鸡西市、鸡东市、佳木斯市、密山市、虎林等地,共采集猪血清样本663份。采样猪群包括公猪、母猪、仔猪、育肥猪。采集的猪场包括规模化猪场、小规模的猪场及散养户。应用ELISA方法对该省东部地区猪蓝耳病的流行情况进行血清学调查,结果显示:663份血清中,其中,134头为阳性,阳性率为20.21%;529头为阴性,阴性率为79.79%;S/P值大于2.0的26头,占总阳性比率的19.40%。     

规模化猪场蓝耳病血清学调查

蓝耳病是影响规模化猪场繁殖性能的主要传染性疾病,笔者在江苏省6个年产万头商品猪的规模化猪场猪群中随机采集297头种猪及仔猪血清进行蓝耳病血清学调查,使用IDEXX酶联免疫吸附诊断试剂盒,检测蓝耳病病毒抗体。结果表明,2个未免疫接种老猪场猪群抗体阳性率52%,断奶仔猪群阳性率39.1%;2个免疫接种老猪场猪群抗体阳性率94%,断奶仔猪群阳性率89.8%。而较新猪场的种猪群阳性率28.6%,断奶仔猪群阳性率5%。     

Isolation and serological survey of Salmonella in pigs in Japan

A total of 267 fecal and serum samples collected from individual pigs reared on a Salmonella-positive farm were subjected to bacteriological and serological examinations of Salmonella. Salmonella was isolated from 47 pigs (17.6%) and prevalence of antibody to lipopolysaccharide (LPS) of S. Typhimurium, which was partly common to S. O4, 12: d: -, was observed in 90 pigs (33.7%). Salmonella was isolated from 26 (28.9%) of 90 antibody-positive pigs and 21 (11.9%) of 177 antibody-negative pigs. Twenty-one of 36 pigs (58.3%) positive for S. O4, 12: d: -, five of 10 pigs (50.0%) positive for S. Havana, and none for S. Anatum had antibodies. Thus, seropositive rates were higher than isolation-positive rates, and antibody prevalence was associated with serovars of the isolates. Then, we analyzed antibody prevalence among pigs on Japanese pig farms. The antibodies to LPS of S. Typhimurium were found in 195 of 1,498 pigs (13.0%) and in at least one serum sample on 35 of 52 farms (67.3%). Our results indicate that Salmonella does not seem to be so prevalent in pigs though it is widely prevalent among pig farms.     

Comparison of Different Commercial Serological Tests for the Detection of Toxoplasma gondii Antibodies in Serum of Naturally Exposed Pigs

Toxoplasma gondii is the aetiological agent of the zoonotic disease toxoplasmosis and transmitted among other ways by chemically and physically untreated, that is, raw pork to humans. The detection of Toxoplasma gondii is impossible by currently practiced meat inspection, but serological tests can be used to detect Toxoplasma gondii antibodies in pig herds and can consequently be helpful to identify potentially contaminated pork. Therefore, appropriate serological tests are required. In this study, serum samples of 1368 naturally exposed slaughter pigs from 73 Austrian farms were collected. Serum samples of at least 16 slaughter pigs per farm were tested. The prevalence of Toxoplasma gondii antibodies in serum was measured by a commercial available modified agglutination test (MAT) and compared to three different commercial available enzyme‐linked immunosorbent assays (ELISA). The MAT detected 6.5%, ELISA I 6.7%, ELISA II 4.8% and ELISA III 4.3% of the pigs as Toxoplasma gondii antibody positive. The agreement, according to the kappa coefficient (κ), was substantial between the MAT and ELISA I (κ = 0.62), II (κ = 0.64) and III (κ = 0.67). A better agreement was determined between ELISA I and II (κ = 0.715), ELISA I and III (κ = 0.747) and ELISA II and III (κ = 0.865). At least one pig per farm was detected Toxoplasma gondii antibody positive in 17 (23.3%) farms by the MAT, 26 (35.6%) farms by ELISA I, 16 (21.9%) farms by ELISA II and 11 (15.1%) farms by ELISA III. Pig farms with a high number of Toxoplasma gondii antibody‐positive pigs or high antibody titres were identified by all of the four used serological tests. Concerning the occurrence of Toxoplasma gondii antibodies in Austrian pig farms, a monitoring and surveillance programme would be reasonable to find high‐risk farms.     

Salmonella typhimurium DT104 septicaemia with meningitis in neonatal piglets.

Clinical salmonellosis in pigs in the Netherlands usually manifests itself as diarrhoea. In finishing pigs this is sometimes accompanied by peracute mortality, mainly in the last month of the finishing period. This is the first report describing Salmonella Typhimurium DT104 infection of 1-week-old suckling piglets in the Netherlands. The piglets showed nervous symptoms and died. The clinical symptoms, gross pathology, histopathological, bacteriological and phagetyping results are presented as well as the antimicrobial resistance pattern. This case is not only important as an extension of the clinical syndrome of salmonellosis in pigs in the Netherlands, but also because of the risk of human infection after consumption of pork or pork products contaminated with this pathogenic and multiple resistant Salmonella clone.     

Patterns of exposure to Lawsonia intracellularis infection on European pig farms

A serological investigation was made of the patterns of exposure of pigs to Lawsonia intracellularis, the causative agent of proliferative enteropathy (ileitis), on farms in France and Spain. Blood samples from groups of adult female pigs in breeding programmes and from postweaning pigs were monitored, the latter every month for five months, by a L. intracellularis-specific immunofluorescence seroassay. Four of 33 farms monitored in France (12 per cent) and three of 29 farms monitored in Spain (10.3 per cent) remained free of clinical signs and seronegative throughout the study. The postweaning pigs on all of the remaining French farms and on 20 of the 26 remaining Spanish farms had a pattern of infection characterised by seroconversion in the grower period, generally between eight and 16 weeks of age. The seroprevalence in these groups ranged from 8 to 20 per cent. On all of these farms at least 15 per cent of the breeding females tested were seropositive, and the farms were under similar management systems, with a continuous flow of pigs or between buildings on one site, so-called 'one site, farrow-to-finish'. On the six remaining Spanish farms, under two management groups, a multiple-site system was used, with the piglets being separated from the adults at weaning and moved to a separate location. On three of these farms, the pattern of infection was characterised by seroconversion later in the finisher period, at between 16 and 20 weeks of age, and none of the breeding females was seropositive. On the three other multiple-site farms the pattern of infection resembled that on the one-site farms. On all of the farms, the seroconversion of groups of pigs was frequently associated with clinical or subclinical signs of ileitis.     

规模化种猪场猪瘟免疫情况调研

猪瘟是由猪瘟病毒引起的一种以高热、出血为主要特征的烈性、高度接触性传染病,至今仍在中国广泛流行。接种疫苗是防控该病发生的最根本的方法,为了查清山西省规模化种猪场猪瘟疫苗免疫情况,课题组采用ELISA试剂盒,对8个地市42个规模化种猪场465头哺乳猪、456头保育猪、436头育肥猪、419头母猪进行了猪瘟免疫抗体检测。查明了哺乳猪抗体阳性率平均为70.11%,保育猪抗体阳性率平均为40.57%,育肥猪抗体阳性率平均为50.22%,母猪抗体阳性率平均为69.69%,证明被检猪群免疫抗体不理想,尤其是保育猪抗体水平较低。同时对12个规模化种猪场,4类不同免疫方法免疫猪瘟疫苗的133头哺乳猪、110头保育猪、105头育肥猪、135头母猪进行了免疫抗体检测。结果表明:乳猪在吃奶前超免猪瘟高效细胞苗6头份,21日龄时二免猪瘟高效细胞苗4头份,60日龄三免猪瘟高效细胞苗2头份,母猪产后21 d跟胎免疫效果最好,使保育猪免疫抗体阳性率达到89.29%,有些猪场使用该方法免疫后,使保育猪的死亡率有了明显的降低,生长发育逐渐走向正常;而采用仔猪在断奶后28日龄至35日龄时首免猪瘟普通细胞苗4头份,60日龄二免猪瘟普通细胞苗2头份,母猪产后28 d跟胎免疫的方法效果最差,不宜推广应用。通过对山西一些规模化种猪场猪瘟免疫情况的调研,基本查清了规模化种猪场猪瘟免疫效果和最佳免疫方法,可为养猪场防控猪瘟提供依据。     

The effect of colostral immunity on the active formation of antibodies in pigs after the administration of inactivated vaccine against Aujeszky's disease

In a large herd of pigs where a trial was performed to cure the animals from Aujeszky's disease (AD) by applying to all animals an inactivated vaccine, a post-vaccination antibody response was studied in piglets coming from the sows that were vaccinated several times. When the piglets were vaccinated at the age of eight weeks (the average virus-neutralizing titer (VNT) of colostral antibodies was 1:11.4) and revaccinated at the age of 11 weeks, 73% of the forty-five animals (examined at the age of 17 weeks) did not have any virus-neutralizing (VN) antibodies in the blood serum. After the third vaccination dose (at the age of 17 weeks), 11% of piglets did not have any VN antibodies if they were examined at the age of 22 weeks (the average antibody VNT was 1:15.3). Applying the ELISA procedure, the antibodies were demonstrated in the sera of all piglets after three vaccination doses. Shifting the time intervals of vaccination (at the age of 8, 13 and 19 weeks), the VN antibodies were found out after three vaccination doses in the sera of all piglets examined at the age of 23 weeks (the average VNT was 1:56.4). After three vaccination doses at the age of 12, 17 and 23 weeks, the VN antibodies were also demonstrated in all piglets at the age of 27 weeks (the average VNT was 1:208).     

2016年-2017年新疆规模化猪场伪狂犬病抗体检测

为了解新疆地区不同规模化猪场伪狂犬病病毒(PRV)野毒感染情况,采用ELISA对2016年4月-2017年12月来自67个不同规模化猪场的2 516份血清进行了PRV-gE抗体血清学调查。结果显示,血清总阳性率为34.02%,2016年和2017年阳性率分别为17.77%和39.52%;哺乳仔猪、保育猪、育肥猪、后备母猪、妊娠母猪、哺乳母猪、公猪总阳性率分别为26.07%、22.35%、35.38%、32.70%、33.13%、31.06%和31.71%;猪场总阳性率为82.09%,北疆、南疆和东疆猪场阳性率分别为78.57%、100.00%和100.00%。结果表明,2016年-2017年新疆地区猪群中PRV感染率明显上升,南疆与东疆地区阳性率显著高于北疆地区,各个阶段猪群阳性率均显著升高,母猪与公猪感染加重是伪狂犬病持续存在的重要原因,对其进行防控与净化刻不容缓。     

Progression of Mycoplasma hyosynoviae infection in three pig herds. Development of tonsillar carrier state, arthritis and antibodies in serum and synovial fluid in pigs from birth to slaughter

In this investigation, natural infection with Mycoplasma hyosynoviae was followed in groups of individual pigs in three different herds with regard to occurrence of tonsillar carrier state, clinical arthritis and development of antibodies in serum and in synovial fluid. Antibodies were detected by a polyclonal enzyme-linked immunosorbent assay (ELISA) developed for experimental use. The infection with M. hyosynoviae progressed very differently in the three herds investigated. In one herd, the infection was apparently limited to adult pigs. In a second herd, all pigs became tonsillar carriers of M. hyosynoviae, but no mycoplasma-related arthritis nor any serological response was demonstrated within the growing-finishing period. In the third herd investigated, tonsillar infection was detected in all pigs, clinical cases of M. hyosynoviae arthritis followed and a moderate serological response was observed in some, but not all, pigs. In all three herds, M. hyosynoviae infection was carried in the tonsils of the adult pigs, but it was only occasionally transmitted from sows to piglets. Maternal antibodies were transferred to the piglets and persisted for approximately 8-12 weeks. After weaning, some pigs became infected before 20 weeks of age, while others did not. In the majority of cases, the tonsillar infection was established from 11 weeks of age or older. A latent tonsillar infection was present for a period of several weeks within the group of investigated pigs before cases of generalized infection and arthritis were seen. In some cases, generalization of M. hyosynoviae infection in the blood and in joints was observed in spite of the detection of an active serological response a few weeks earlier. The present work suggests that generalization of the infection and development of arthritis may depend on age, immunity, virulence factors and/or infection pressure; in some herds maybe combined with certain triggering mechanisms such as stress and lowered general resistance.     

Dynamics of influenza A virus infections in permanently infected pig farms: evidence of recurrent infections,circulation of several swine influenza viruses and reassortment events

Concomitant infections by different influenza A virus subtypes within pig farms increase the risk of new reassortant virus emergence. The aims of this study were to characterize the epidemiology of recurrent swine influenza virus infections and identify their main determinants. A follow-up study was carried out in 3 selected farms known to be affected by repeated influenza infections. Three batches of pigs were followed within each farm from birth to slaughter through a representative sample of 40 piglets per batch. Piglets were monitored individually on a monthly basis for serology and clinical parameters. When a flu outbreak occurred, daily virological and clinical investigations were carried out for two weeks. Influenza outbreaks, confirmed by influenza A virus detection, were reported at least once in each batch. These outbreaks occurred at a constant age within farms and were correlated with an increased frequency of sneezing and coughing fits. H1N1 and H1N2 viruses from European enzootic subtypes and reassortants between viruses from these lineages were consecutively and sometimes simultaneously identified depending on the batch, suggesting virus co-circulations at the farm, batch and sometimes individual levels. The estimated reproduction ratio R of influenza outbreaks ranged between 2.5 [1.9-2.9] and 6.9 [4.1-10.5] according to the age at infection-time and serological status of infected piglets. Duration of shedding was influenced by the age at infection time, the serological status of the dam and mingling practices. An impaired humoral response was identified in piglets infected at a time when they still presented maternally-derived antibodies.     

A field evaluation of Nobi-Vac atrophic rhinitis vaccine

A field trial was carried out with Nobi-Vac AR vaccine in 11 swine breeding herds. One herd suffered from severe B. bronchiseptica (BB+) rhinitis in piglets, while no clinical Atrophic Rhinitis (AR) was observed. Ten herds were described as AR problem herds, because clinical AR was observed for at least 1 year in spite of the fact that medication of sows and piglets was carried out and adequate housing and management systems were available in most herds. BB+ was isolated in 9 of these 10 farms and the AR pathogenic P. multocida (PM+) in all these farms. The trial started when piglets were born of sows which had been vaccinated 3 times. All piglets between 7 and 10 weeks old were examined clinically for AR. Nasal swabs from 20 pigs on each farm were screened bacteriologically for BB+ and PM+ every other month. A severe reoccurrence of AR was found in 2 of the 10 AR problem farms. In these 2 herds the 'all in-all out' system was not applied in farrowing and weaning houses. In the 8 other AR problem herds the percentage of pigs with AR decreased significantly below the average level of 1% per year. The percentage of pigs infected with BB+ and PM+ also decreased significantly. PM+ was significantly related to the percentage of clinical AR piglets. Seasonal effects contributed to a minor extent to the percentage of AR piglets. After 2.5 years of this trial PM+ could not be isolated during the last 12 months in six of the remaining AR problems herds. The conclusion is that Nobi-Vac AR vaccination of the sow gave protection against clinical AR in piglets on those farms (80%) which provided adequate housing and management systems.     

Effect of maternal antibodies and pig age on the antibody response after vaccination against Glässers disease

The influence of age and maternal antibodies on the development and duration of postvaccinal antibody response against Glässer’s disease were investigated. Pigs born to immune (MDA-positive) and non-immune (MDA-negative) sows were vaccinated with inactivated vaccine. Vaccination was done according to three different protocols: at 1 and 4, at 2 and 5 or at 4 and 7 weeks of age. There were also two control groups for MDA-negative and MDA-positive pigs. The level of Haemophilus parasuis (Hps) specific antibodies were determined using commercial ELISA test. No serological responses were seen in any of the groups after the first vaccination. Maternally derived antibodies (MDA) against Hps were above the positive level until approximately 3 weeks of life in MDA-positive pigs. In those pigs the strongest postvaccinal humoral response was observed in piglets vaccinated at 4 and 7 weeks of age. In the remaining MDA-positive piglets only slight seroconversion was noted but levels of antibodies never exceeded values considered as positive. All MDA-negative pigs produced Hps-specific antibodies after the second vaccination. The results of the present study indicated that MDA may alter the development and duration of active postvaccinal antibody response. Age of pigs at the moment of vaccination was not associated with the significant differences in the magnitude of antibody response, however influenced the kinetics of decline of Hps-specific antibodies.