Porcine Circovirus Type 2 Antibodies Research Articles

Synergistic pathogenicity in sequential coinfection with Mycoplasma hyorhinis and porcine circovirus type 2

To investigate the synergistic pathogenicity of Mycoplasma hyorhinis (Mhr) with porcine circovirus type 2 (PCV2), thirty 35-day-old piglets were randomly distributed to six groups (n=5 each): Mhr/PCV2 (group 1, inoculated with Mhr and PCV2 1 week later), PCV2/Mhr (group 2, inoculated with PCV2 and Mhr 1 week later), Mhr–PCV2 (group 3, inoculated with PCV2 and Mhr concurrently), singular PCV2 group (group 4), singular Mhr group (group 5), and a uninfected control group (group 6). Mild transient lethargy, fever, coughing, inappetence, and decreased daily weight gain were observed in all dual-infected groups and the singular Mhr-infected group. There were significantly higher levels of PCV2 and Mhr antibodies, larger amounts and wider range of tissue distribution of PCV2 antigens and nucleic acids in the dual-infected groups compared to the single-infected and control groups. PCV2 and Mhr dual-infection resulted in significantly more severe macroscopic and microscopic lung lesions and wider PCV2 DNA distribution compared with piglets infected with PCV2 alone. Cytokine detection showed a significant change in tumor necrosis factor-α, interleukin-2, and interleukin-6 levels in the infected groups, especially in the Mhr–PCV2 group, compared with the control group. Hence, Mhr potentiated the severity of PCV2-associated lung lesions, increased the amount and distribution of PCV2 DNA in tissues, and increased the incidence of porcine respiratory disease.

Influence of repeated porcine circovirus type 2 vaccination on sows’ immune response

In the last decade, the impact of the porcine circovirus type 2 (PCV-2)-associated diseases in pigs has consistently decreased due to the extensive application of PCV-2 vaccines in sows and...

Enhanced immunoassay for porcine circovirus type 2 antibody using enzyme-loaded and quantum dots-embedded shell–core silica nanospheres based on enzyme-linked immunosorbent assay

Boosting the detection sensitivity of enzyme-linked immunosorbent assay (ELISA) is significant to the early clinical diagnosis of various diseases. Here, we developed a versatile immunosensor using silica nanospheres as carriers for sensitive detection of porcine circovirus type 2 (PCV2) antibody. With HRP enzyme covalently immobilized on the silica nanospheres and CdSe nanocrystals embedded inside, these signal probes were successfully utilized in the sensitive detection of PCV2 antibody by ELISA, fluorometry and square-wave voltammetry (SWV). To further demonstrate the performance of the immunosensor, Human IgG (HIgG) was used as a model analyte. Since more HRP and CdSe QDs were loaded, 5-, 200- and 400-fold enhancements in amplified ELISA, fluorometry and voltammetry responses for HIgG could be achieved compared to conventional ELISA. The respective detection limits of theses methods for HIgG were 3.9, 0.1 and 0.05 ng mL−1 with a RSD below 5% for amplified ELISA, fluorescence and SWV measurements. Additionally, a 100-fold improvement was obtained in the detection sensitivity for PCV2 antibody immunoassay. The versatile immunosensor exhibits good sensitivity, stability and reproducibility, suggesting its potential applications in clinical diagnostics.

Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems.

Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.

Development of a surface plasmon resonance biosensing approach for the rapid detection of porcine circovirus type2 in sample solutions.

A sensitive and label-free analytical approach for the detection of porcine circovirus type 2 (PCV2) instead of PCV2 antibody in serum sample was systematically investigated in this research based on surface plasmon resonance (SPR) with an establishment of special molecular identification membrane. The experimental device for constructing the biosensing analyzer is composed of an integrated biosensor, a home-made microfluidic module, and an electrical control circuit incorporated with a photoelectric converter. In order to detect the PCV2 using the surface plasmon resonance immunoassay, the mercaptopropionic acid has been used to bind the Au film in advance through the known form of the strong S-Au covalent bonds formed by the chemical radical of the mercaptopropionic acid and the Au film. PCV2 antibodies were bonded with the mercaptopropionic acid by covalent -CO-NH- amide bonding. For the purpose of evaluating the performance of this approach, the known concentrations of PCV2 Cap protein of 10 µg/mL, 7.5 µg/mL, 5 µg/mL, 2.5 µg/mL, 1 µg/mL, and 0.5 µg/mL were prepared by diluting with PBS successively and then the delta response units (ΔRUs) were measured individually. Using the data collected from the linear CCD array, the ΔRUs gave a linear response over a wide concentration range of standard known concentrations of PCV2 Cap protein with the R-Squared value of 0.99625. The theoretical limit of detection was calculated to be 0.04 µg/mL for the surface plasmon resonance biosensing approach. Correspondingly, the recovery rate ranged from 81.0% to 89.3% was obtained. In contrast to the PCV2 detection kits, this surface plasmon resonance biosensing system was validated through linearity, precision and recovery, which demonstrated that the surface plasmon resonance immunoassay is reliable and robust. It was concluded that the detection method which is associated with biomembrane properties is expected to contribute much to determine the PCV2 in sample solutions instead of PCV2 antibody in serum samples quantitatively.

Effect of porcine circovirus type 2 (PCV2) load in serum on average daily weight gain during the postweaning period

Porcine circovirus type 2 (PCV2) is a ubiquitous virus that mainly affects nursery and fattening pigs causing systemic disease (PCV2-SD) or subclinical infection. A characteristic sign in both presentations is reduction of average daily weight gain (ADWG). The present study aimed to assess the relationship between PCV2 load in serum and ADWG from 3 (weaning) to 21 weeks of age (slaughter) (ADWG 3-21). Thus, three different boar lines were used to inseminate sows from two PCV2-SD affected farms. One or two pigs per sow were selected (60, 61 and 51 piglets from Pietrain, Pietrain×Large White and Duroc×Large White boar lines, respectively). Pigs were bled at 3, 9, 15 and 21 weeks of age and weighted at 3 and 21 weeks. Area under the curve of the viral load at all sampling times (AUCqPCR 3-21) was calculated for each animal according to standard and real time quantitative PCR results; this variable was categorized as “negative or low” (<104.3 PCV2 genome copies/ml of serum), “medium” (≥104.3 to ≤105.3) and “high” (>105.3). Data regarding sex, PCV2 antibody titre at weaning and sow parity was also collected. A generalized linear model was performed, obtaining that paternal genetic line and AUCqPCR 3-21 were related to ADWG 3-21. ADWG 3-21 (mean±typical error) for “negative or low”, “medium” and “high” AUCqPCR 3-21 was 672±9, 650±12 and 603±16g/day, respectively, showing significant differences among them. This study describes different ADWG performances in 3 pig populations that suffered from different degrees of PCV2 viraemia.

A recombinant porcine circovirus type 2 expressing the VP1 epitope of the type O foot-and-mouth disease virus is infectious and induce both PCV2 and VP1 epitope antibodies

Porcine circovirus type 2 (PCV2) is the etiological agent of postweaning multisystemic wasting syndrome, a disease that causes huge economic damage in swine industry. A recombinant PCV2 expressing the neutralizing VP1 epitope (aa 141-160) of the foot-and-mouth disease virus (FMDV) was rescued using an infectious cloning technique. The PCV2 antigen and FMDV-VP1 antigenic epitope of the cloned strain recPCV2-CL-VP1 were confirmed by an immunoperoxidase monolayer assay (IPMA) and a capture enzyme-linked immunosorbent assay (ELISA). The morphological features of the recPCV2-CL-VP1 were not discernibly different from those of its parental strain (PCV2-CL). However, the recombinant virus could be differentiated from its parental virus by PCR and capture ELISA. The recPCV2-CL-VP1 was demonstrated to replicate stably in PK-15 cells through ten passages. An infection experiment using BALB/c mice showed that both recPCV2-CL-VP1 and PCV2-CL could replicate in the mice, cause various pathological changes, and induce a high level of anti-Cap antibodies. The recombinant virus emulsified with Freund's adjuvant was used to immunize BALB/c mice and induced antibodies against the FMDV-VP1 epitope. Hence, the recombinant PCV2 strain, which expressed the neutralizing FMDV-VP1 epitope, provides a valuable platform to develop novel genetic vaccines.

Host genetic heterozygosity and age are important determinants of porcine circovirus type 2 disease prevalence in European wild boar

Emerging and zoonotic diseases are important challenges for veterinary and public health. It is therefore a key issue to assess the relative importance of various factors for disease dynamics and to understand the mechanisms behind these factors and interactions. Here, we evaluate the influence of a number of demographic and genetic factors on porcine circovirus type 2 (PCV2) antibody prevalence in the European wild boar (Sus scrofa). We measured PCV2 blood serum antibody levels of 462 randomly sampled wild boars from a cross-border area in the Netherlands and western parts of Germany in a 3-year period. These samples were also genotyped using a randomly selected genome-wide 351 SNP assay. Generalized linear mixed model analysis shows that wild boar PCV2 antibody status is determined by age and genetic heterozygosity, with an idiosyncratic influence of the year of sampling. In contrast, sex, population membership and domestic hybrid status did not significantly affect PCV2 antibody status. The observed positive relationship between PCV2 antibody status and age is most likely caused by cumulative exposure and PCV2-typical intracellular hiding behaviour. The observed positive relationship between wild boar genetic heterozygosity and PCV2 antibody status could be attributed to disappearance of relatively inbred (low-heterozygosity) individuals. This finding suggests that PCV2 can act as a selective force in wild boar populations and that disease mortality can be mediated by host heterozygosity.

Effect of transfer factor on immune efficacy of PCV2 subunit vaccine

To investigate the effect of transfer factor(TF) on the immune efficacy of porcine circovirus type 2(PCV2) subunit vaccine,spleen from healthy porcine was grinded and the homogenate was frozen-thawed six times.TF was obtained after the homogenate was centrifuged and ultrafiltered.After physical and chemical inspection,TF was added to PCV2 subunit vaccine(Cap vaccine),named TF-Cap vaccine.TF-Cap vaccine and Cap vaccine were used to immunize mice with physiological saline as control.The mice were separated into two groups: one-time vaccination and two-time vaccination.Blood was collected at 10-d interval through caudal vein after the first vaccination and PCV2 antibody was tested using indirect enzyme-linked immuno-sorbent assay.The result showed that the concentration of the ultrafiltered TF is 1.83 mg/mL.Detection of PCV2 specific antibody showed that there are significant differences(P0.01) between vaccine group and control group while there is no significant differences(P0.05) among vaccine groups.However,PCV2 antibody titer in mice immunized TF-Cap vaccine only once was comparable to mice immunized Cap-vaccine twice,indicating PCV2 subunit vaccine with TF can enhance the antibody production.

Anti-porcine circovirus type 2 (PCV2) antibody placental barrier leakage from sow to fetus: impact on the diagnosis of intra-uterine PCV2 infection

Dear Editor,Fetuses develop antibodies when they are infectedwith porcine circovirus type 2(PCV2).At abortion orbirth,both PCV2 and/or anti-PCV2 antibodies can thenbe detected.Finding anti-PCV2 antibodies in aborted fe-tuses or stillborn piglets is generally used as evidence ofan intra-uterine PCV2 infection.However,a recent anal-ysis of fetuses sent in for PCV2 diagnosis and caesare-an-derived,colostrum-deprived pigs revealed that low

Pigs naturally exposed to porcine circovirus type 2 (PCV2) generate antibody responses capable to neutralise PCV2 isolates of different genotypes and geographic origins

Porcine circovirus type 2 (PCV2) is the essential infectious agent for PCV2-systemic disease (PCV2-SD, formerly known as postweaning multisystemic wasting syndrome) and other pathological conditions. Recent studies indicated antigenic variability amongst different PCV2 isolates and suggested that single amino acid changes within the capsid protein determine differences in the level of neutralization by specific monoclonal antibodies. The objective of the present study was to examine the cross-reactivity of PCV2 antibodies induced in the context of a natural infection against different PCV2 isolates belonging to genotypes PCV2a and PCV2b. Sera taken from several farms from animals of varying health status (PCV2-SD and age-matched healthy pigs and a set of slaughter-aged animals) were assayed for neutralizing activity against four PCV2 isolates from both predominant genotypes (PCV2a and PCV2b) and of differing geographic origins (Europe and North-America). Results showed that most of studied pigs (79 out of 82) contained neutralizing antibodies (NA) able to neutralize all four studied viral strains. Overall, pigs had significantly higher NA titres against PCV2a than against PCV2b (P < 0.001). Accordingly, studied serums were able to better neutralize Burgos390L4 and Stoon-1010 strains (PCV2a) than L-33-Sp-10-54 and MO/S-06 strains (PCV2b) (P < 0.001). No differences between capabilities of seroneutralization of viruses from different geographic origin were observed. Present data suggests that sequence differences between PCV2 isolates translate to functional antigenic differences in viral neutralization in vivo.

Research of immune response in unvaccinated piglets against porcine circovirus type 2

Infection of pigs caused by porcine circovirus type 2 (PCV2) is present throughout the world. The aim of this study was to determine, based on the follow-up of the class IgG titers in piglets and fattening pigs, the duration of colostral immunity against infections in pigs caused by PCV2. The study included 28 piglets, whose titer was determined by using indirect ELISA test in intervals up to 110 days of age. The observed average values of IgG in piglets aged 21 days (8.47 log2) and 35 days (6.69 log2) in our surveys indicate very high titers of maternal antibodies, and that the piglets at this age are, to some extent, protected from infection caused by PCV2. Absence of the specific anti PCV2 antibodies was determined in 14.29% (21 days old) and 15.38% (35 days old) piglets at this age. The average titer of antibodies, specific for PCV2, in blood serum of piglets on the 50th day decreased to 3.74 log2, what indicates that in the majority of piglets (61.54%) catabolism of colostral antibodies occurred. On the 80th day of the piglets life there were no seropositive specimens, which unambiguously indicates that in this age, there was a complete catabolism of colostral antibodies and the disappearance of passive immunity. In all the 110 days? old fattening pigs, an average antibody titer recorded a sharp rise (12.78 log2). Possible reasons for this sudden increase in PCV2 antibody titers in the blood serum of fattening pigs is is a widespread presence of this infection in our region, as well as the fact that there exists a part of pigs population which is not immune to PCV2 infection. The results of this research are important for choosing the optimal moment for vaccination, considering that high titers of colostral antibodies of class G on the 21st and 35th day of piglets? life have been proven. This data points out to a possible interference with vaccine immunogens in the case of vaccination in this particular age.

Effects of algae-derived β-glucans with zinc on nursery pig growth performance and immune response under commercial conditions (2014)

Effects of algae-derived β-glucans with zinc on nursery pig growth performance and immune response under commercial conditions (2014)

Detection of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in meat juice samples from Polish wild boar (Sus scrofa L.)

PCV2 and PRRSV are two important pathogens of domestic swine. There is considerable evidence that the infection is also present in wild boars. Meat juice provides an alternative to serum for antibody testing, and it has been used in testing for many important porcine infectious diseases. Samples of brachial muscle were collected from 142 wild boars shot in different regions of Poland during the 2006/2007 and 2007/2008 hunting seasons. Meat juice harvested from muscle samples was tested using an ELISA test specific for PCV2 and PRRSV antibodies. Additionally, IgG and IgM antibodies specific for PCV2 were detected in order to estimate the status of the PCV2 infection. Only one of the tested meat juice samples was positive for PRRSV (0.7%), and 68 out of 142 (47.9%) samples were positive for PCV2. Of the positive animals, 4 (2.8%) had an antibody profile suggesting active infection, 2 (1.4%) early active infection, and 62 (43.7%) late infection. Also, a lack of association between the age of the animals and the presence of antibodies related to the infection was noticed.

Humoral response and colostral antibody transfer following ‘one-dose’ pre-mating vaccination of sows against porcine circovirus type-2

The objective of this study was to investigate the presence of porcine circovirus type 2 (PCV2) DNA and antibody to the virus in the serum and colostrum of sows vaccinated prior to mating and in their offspring. Seventy-seven sows were randomly distributed into vaccinated (V, n=36) and non-vaccinated (NV, n=41) groups. One week before mating, sows were given a PCV2 vaccine (V group) or PBS (NV group) IM. Blood samples were taken from the sows at fixed time-points and colostrum samples were taken at farrowing. Blood samples were also taken from the piglets of the sows at 4weeks of age. The results indicated that vaccination prior to mating elicited a strong, homogeneous humoral response and, in consequence, more homogeneous colostral PCV2 antibody concentrations.

Half-life of porcine antibodies absorbed from a colostrum supplement containing porcine immunoglobulins

Absorption of immunoglobulins (Ig) at birth from colostrum is essential for piglet survival. The objective was to evaluate the half-life of antibodies absorbed in the bloodstream of newborn piglets orally fed a colostrum supplement (CS) containing energy (fat and carbohydrates) and IgG from porcine plasma. Viable piglets (n = 23; 900 to 1,800 g BW) from 6 sows were colostrum deprived and blood sampled and within the next 2 h of life randomly allocated to either control group (n = 9) providing 30 mL of Ig-free milk replacer or a group (n = 14) receiving 30 mL of CS by oral gavage. Piglets were transported to a Biosafety Level 3 facility (Centre de Recerca en Sanitat Animal, Spain) and fed Ig-free milk replacer every 3 to 4 h for 15 d. Survival, weight, plasma IgG content by radial immunodiffusion (RID), and antibodies against porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome (PRRS), Mycoplasma hyopneumoniae (Mhy), and swine influenza virus (SIV) were determined by specific ELISA before treatment administration, at 24 h, and weekly for 56 d. Clinical symptoms were not observed for either group. Mortality index was lower (17 vs. 38%; P < 0.02) and BW higher (17.7 vs. 15.3 kg; P = 0.035) for pigs supplemented with CS than piglets in the control group. At 24 h postadministration, the CS group had a plasma IgG mean of 7.6 ± 0.06 vs. 0.14 ± 0.03 mg/mL for the control group. The IgG levels in the CS group decayed until day 21 when de novo synthesis of IgG was detected in 25% of piglets. Half-life of antibody concentration (HLAC) by RID was 6.2 d. In the CS group, efficiency of PCV2 and PPV antibody transfer was high. For PCV2, all animals remained positive by day 56 and the calculated HLAC was 17.7 d. For PPV, 72.7% of piglets were ELISA positive by day 35 and HLAC was 12.0 d. For PRRS, all piglets remained positive by day 14 and the calculated HLAC was 11.9 d. For Mhy and SIV the calculated HLAC were 8.4 and 3.0 d. In summary, half-life of antibodies derived from blood plasma in the bloodstream of newborn piglets varied from 3.0 to 17.7 d. The study also confirm that antibodies derived from porcine plasma were well absorbed and can be an useful tool for providing protection against several or specific pathogens and can be a good alternative to formulate CS for newborn piglets.

Evaluation of the transmission of porcine circovirus type 2 (PCV-2) genogroups a and b with semen from infected specific-pathogen-free boars

The porcine circovirus type 2 (PCV-2) is associated with several diseases including reproductive failure. This syndrome has been experimentally reproduced twice with two PCV-2 isolates representative of each major PCV-2 genogroup, i.e. PCV-2a and PCV-2b (Cariolet et al., 2002; Rose et al., 2007). In these two previous studies, the sows were infected by intra-uterine inoculation at insemination with 104.3 and 103.18 TCID50 of PCV-2a and PCV-2b, respectively, corresponding to 1.2×1011 and 3×1010 genome copies, respectively.The aim of this present study was to quantify viral shedding in semen from specific-pathogen-free (SPF) boars infected with isolates from the two major PCV-2 genogroups a and b. We studied the transmission of the PCV-2 virus through contaminated semen to SPF sows and their offspring. The four inoculated boars developed sub-clinical PCV-2 infections and PCV-2 genomes were occasionally detected in semen after nasal infection of boars, with up to 1.2×106copies/mL in the sperm-rich fraction. When PCV-2-contaminated semen was inoculated in SPF sows at artificial insemination, the sows and their offspring did not show any signs of PCV-2 infection or PCV-2 antibodies or genomes. In the present study, sows were inoculated with a maximal dose of 1.7×107 viral genome copies, which is lower than the genomic loads (i.e. 1.2×1011 and 3×1010 genome copies) that have been shown to induce reproductive troubles in intra-uterine inoculated sows. Our results together with the previous experiment findings suggest that PCV-2-induced reproductive disorders depend on the infectious dose inoculated to sows by the intra-uterine route.

Evaluation of cell-mediated immune responses against porcine circovirus type 2 (PCV2) Cap and Rep proteins after vaccination with a commercial PCV2 sub-unit vaccine

This study investigated the development of cellular immunity to Porcine circovirus type 2 (PCV2) Cap and Rep proteins in pigs vaccinated with a commercial PCV2 genotype a (PCV2a) based sub-unit vaccine, before and after a heterologous challenge with a PCV2b isolate. At three weeks of age, 20 pigs were inoculated intramuscularly with either the vaccine product (V group, n=9) or phosphate buffered saline solution (PBS) (NV group, n=11). Three weeks after vaccination, pigs were challenged intranasally with PCV2b (V-C and NV-C groups) or PBS (V-NC and NV-NC groups). None of the pigs developed clinical signs during the whole experiment, but all NV-C and 3/5 V-C pigs developed viraemia. Vaccination induced the development IFN-γ-secreting cells in response to the Cap protein of PCV2, which appeared three weeks post-vaccination and increased after challenge. By that time, no significant differences were detected on PCV2 antibody titres between vaccinated and non-vaccinated pigs, although there were significant differences on day 7 post-challenge. PCV2-inoculation induced a cellular response against the Rep protein. Such response was significantly reduced or even absent in PCV2-inoculated pigs that were previously vaccinated (V-C group), presumably as a result of a lower PCV2 replication in vaccinated animals compared to non-vaccinated ones.

Porcine circovirus type 2 infection before and during an outbreak of postweaning multisystemic wasting syndrome on a pig farm in the UK

The presence of porcine circovirus type 2 (PCV-2) and other pathogens before and during an outbreak of postweaning multisystemic wasting syndrome (PWMS) in pigs is evaluated in this study. At...

A Serosurvey for Brucella suis, Classical Swine Fever Virus, Porcine Circovirus Type 2, and Pseudorabies Virus in Feral Swine (Sus scrofa) of Eastern North Carolina

As feral swine (Sus scrofa) populations expand their range and the opportunity for feral swine hunting increases, there is increased potential for disease transmission that may impact humans, domestic swine, and wildlife. From September 2007 to March 2010, in 13 North Carolina, USA, counties and at Howell Woods Environmental Learning Center, we conducted a serosurvey of feral swine for Brucella suis, pseudorabies virus (PRV), and classical swine fever virus (CSFV); the samples obtained at Howell Woods also were tested for porcine circovirus type 2 (PCV-2). Feral swine serum was collected from trapped and hunter-harvested swine. For the first time since 2004 when screening began, we detected B. suis antibodies in 9% (9/98) of feral swine at Howell Woods and <1% (1/415) in the North Carolina counties. Also, at Howell Woods, we detected PCV-2 antibodies in 59% (53/90) of feral swine. We did not detect antibodies to PRV (n=512) or CSFV (n=307) at Howell Woods or the 13 North Carolina counties, respectively. The detection of feral swine with antibodies to B. suis for the first time in North Carolina warrants increased surveillance of the feral swine population to evaluate speed of disease spread and to establish the potential risk to commercial swine and humans.