Classical Swine Fever Virus Antibody Research Articles

Prevalence of antibodies against classical swine fever in two districts of Nepal

This study aimed to investigate the seroprevalence of Classical Swine Fever (CSF), caused by the Classical Swine Fever Virus (CSFV), in the Bhaktapur and Kavrepalanchok districts of Nepal. The primary objective was to determine the prevalence of antibodies against CSFV in the swine population, providing crucial insights for targeted interventions and control strategies. A cross-sectional study was conducted between January and March 2018. The research involved structured questionnaire surveys and blood sample collection from selected farms in the study area. A total of 184 serum samples were collected and subjected to testing using the IDEXX ELISA test kit. The study evaluated seroprevalence by considering variables such as district, sex, age groups, and housing systems. Out of the 184 serum samples collected, 34 were found to be seropositive for CSFV antibodies, resulting in an overall seroprevalence of 18.40%. The study identified significant variations in seroprevalence between different factors, including districts, sexes, age groups, and housing systems. These findings indicate a notable exposure of swine populations in the Bhaktapur and Kavrepalanchok districts to CSFV. This research highlights the considerable risk posed by CSF to the pig populations in the Bhaktapur and Kavrepalanchok districts of Nepal. The identified seroprevalence and variations among different factors emphasize the importance of targeted interventions and control strategies in these regions. The findings serve as a foundation for informed decision-making to mitigate the impact of CSF, safeguard swine health, and protect the livelihoods of those dependent on the swine industry. This study was made possible with the support of the Zoonosis Control Project under the National Animal Science Research Institute, Nepal Agriculture Research Council.

Identification of novel monoclonal antibodies specific for the conserved epitopes in the E2 protein of genotype 2 classical swine fever virus: implication for differential diagnosis

Since the last century, the spread of the genotype 2 classical swine fever virus (CSFV) has caused significant issues for the pig breeding industries. Ideal strategies for controlling CSFV include vaccination and keeping farms free of CSFV. For vaccination, several attenuated CSFV viruses originating from genotype 1 are widely used; for the latter, accurate diagnosis is required for detection of the CSFV infection. Nucleic acid testing for CSFV usually uses tonsil samples, which requires an inconvenient sampling operation that injures pigs. Commercial serological tests for CSFV antibodies or antigens are unable to distinguish the genotype for originating virus. In this study, 20 mAbs were developed from the mice hybridoma cells. Four of the mAbs were identified to have the ability to only recognize the peptides derived from sub-genotype 2.1 strain, and two of them, MM1 and MM5, were further studied to identify critical binding sites (epitopes) on the E2 protein of CSFV. A total of 353 genotype 2 collections were made worldwide in GeneBank, 90.9% of which contained MM1 or MM5 epitopes. Moreover, 95.1% of sub-genotype 2.1 isolations contained MM5 epitope. Therefore, MM1 and MM5 have the potential to be developed as a diagnostic tool for detection of genotype 2 virus antigen by indirect ELISA or antibodies by competitive ELISA.

A prospective CSFV-PCV2 bivalent vaccine effectively protects against classical swine fever virus and porcine circovirus type 2 dual challenge and prevents horizontal transmission

Classical swine fever virus (CSFV) infection leading to CSF outbreaks is among the most devastating swine diseases in the pig industry. Porcine circovirus type 2 (PCV2) infection, resulting in porcine circovirus-associated disease (PCVAD), is also a highly contagious disease affecting pig health worldwide. To prevent and control disease occurrence, multiple-vaccine immunization is necessary in contaminated areas or countries. In this study, a novel CSFV-PCV2 bivalent vaccine was constructed and demonstrated to be capable of eliciting humoral and cellular immune responses against CSFV and PCV2, respectively. Moreover, a CSFV-PCV2 dual-challenge trial was conducted on specific-pathogen-free (SPF) pigs to evaluate vaccine efficacy. All of the vaccinated pigs survived and showed no clinical signs of infection throughout the experimental period. In contrast, placebo-vaccinated pigs exhibited severe clinical signs of infection and steeply increased viremia levels of CSFV and PCV2 after virus challenge. Additionally, neither clinical signs nor viral detections were noted in the sentinel pigs when cohabitated with vaccinated-challenged pigs at three days post-inoculation of CSFV, indicating that the CSFV-PCV2 bivalent vaccine completely prevents horizontal transmission of CSFV. Furthermore, conventional pigs were utilized to evaluate the application of the CSFV-PCV2 bivalent vaccine in field farms. An adequate CSFV antibody response and a significant decrease in PCV2 viral load in the peripheral lymph nodes were observed in immunized conventional pigs, suggesting its potential for clinical application. Overall, this study demonstrated that the CSFV-PCV2 bivalent vaccine effectively elicited protective immune responses and the ability to prevent horizontal transmission, which could be a prospective strategy for controlling both CSF and PCVAD in commercial herds.

A Novel Blocking Enzyme-Linked Immunosorbent Assay Based on a Biotinylated Nanobody for the Rapid and Sensitive Clinical Detection of Classical Swine Fever Virus Antibodies.

Monoclonal and polyclonal antibodies are mostly used for the development of traditional enzyme-linked immunosorbent assays (ELISAs), but the use of certain conventional antibodies may be limited by their low yield, the difficulty of their isolation, and their high cost. Heavy-chain antibodies derived from camelids with naturally missing light chains can overcome these deficiencies and are an excellent alternative to conventional antibodies. In this study, a nanobody (Nb)-AviTag fusion protein was constructed, and the feasibility of its use as a high-sensitivity probe in a blocking ELISA (bELISA) for classical swine fever virus (CSFV) was investigated. The CSFV E2 recombinant protein expressed by the CHO expression system exhibited good reactogenicity and immunogenicity and induced the production of high CSFV antibody levels in rabbits. Three different clones of Nbs were successfully isolated using a phage display system in alpaca, and an Nb1-AviTag fusion protein was successfully expressed using an Escherichia coli expression system. The purified Nb1-AviTag fusion protein was then biotinylated in vitro to obtain Nb1-biotin. A novel bELISA was developed for the detection of CSFV antibodies in clinical serum using Nb1-biotin as a probe. The cutoff value of bELISA was 32.18%, the sensitivity of bELISA was higher than that of the bELISA kit with IDEXX antibody, and the coincidence rate was 94.7%. A rapid, low-cost, highly sensitive and highly specific CSFV E2 antibody-based bELISA method was successfully established and can be used for the serological evaluation of CSFV E2 subunit vaccines and the ELISA-based diagnosis of CSFV infection. IMPORTANCE Currently, the epidemic situation of classical swine fever (CSF) is sporadic, and cases of atypical swine fever are on the rise in China. Therefore, it is necessary to accurately eliminate suspected cases by using highly sensitive and specific diagnostic techniques. In our study, a rapid, low-cost, highly sensitivity, highly reliable and reproducible, and highly specific classical swine fever virus (CSFV) E2 antibody-based blocking ELISA method was successfully established by using the phage display system and the Nb1-AviTag fusion expression platform. It provides a new technique for serological evaluation of CSFV vaccines and ELISA-based diagnosis of CSFV infection.

Seroprevalence of antibodies to classical swine fever virus and porcine reproductive and respiratory syndrome virus in healthy pigs in Hunan Province, China.

Classical swine fever (CSF) and porcine reproductive and respiratory syndrome (PRRS) are responsible for major economic losses and represent a threat to the swine industry worldwide. Routine surveillance serology for CSF and PRRS viruses is critical to maintaining the health status of sow farms in Hunan Province, which is one of the top pig production provinces in China. The aim of our study was to investigate the serological statistics of CSF virus (CSFV) and PRRS virus (PRRSV) in Hunan Province. The cohort serum samples were collected from vaccinated and unvaccinated pigs. Our findings showed that the average rates of CSFV and PRRSV antibody seropositivity were 82.2% (95% CI: 80.1-84.3) and 84.8% (95% CI: 82.5-87.1), respectively, in the immunized group and that these rates were higher than those in the unvaccinated group (58.6% for CSFV and 47.8% for PRRSV). Additionally, the level of CSFV antibody in piglet serum declined gradually with age, whereas PRRSV-specific antibody level increased initially (1 to 2 weeks old) and then declined with age (2 to 4 weeks old). In summary, we investigated the difference in CSFV/PRRSV antibody levels among piglets at various weeks old (1 to 4 weeks) to further establish the duration of maternal immunity in piglets. In addition, routine monitoring of CSFV/PRRSV antibodies in immunized pigs was carried out to evaluate the efficacy of vaccination.

Development and application of a high-sensitivity immunochromatographic test strip for detecting classical swine fever virus antibodies.

Classical swine fever (CSF) is caused by classical swine fever virus (CSFV) and has led to huge economic losses in the pig industry worldwide. Although vaccination and other control measures have been carried out, it is essential to establish a rapid and valid method for CSF vaccination monitoring and clinical diagnosis. The CSFV E2 protein has been widely used as a major antigen for antibody detection. It is important to improve the affinity between the E2 protein and CSFV antibodies to improve the performance of the detection method. In this study, a recombinant E2 extracellular protein (amino acids 1-331) with a native homodimer conformation and high affinity for the anti-CSFV-E2 monoclonal antibody WH303 was expressed using a Bac-to-Bac baculovirus expression system. A novel immunochromatographic test strip based on the recombinant CSFV E2 protein was developed for CSFV antibody detection. The sensitivity of this strip for detecting CSFV standard-positive serum was 1:102400, four times higher than that of the previously developed CnC2 test strip. No cross-reactivity with antibodies of other swine viruses was observed. Detection of clinical swine serum samples (n=813) demonstrated that the agreements of this E2 test strip with three commercial ELISA kits were 97.17% (790/813), 95.94% (780/813) and 93.73% (762/813), respectively. Our data indicate that a novel E2 test strip with enhanced sensitivity has been developed and can be applied for clinical sample detection, providing a new, powerful and simple approach for CSFV antibody monitoring.

Evidence of transboundary transmission of viral pathogens through pigs transported from Myanmar to Mizoram, India

In Mizoram, pigs used to migrate through the porous international border from Myanmar. It is speculated that the migrated animals may act as potential carriers of various microbial pathogens. But so far no scientific evidence is available in this aspect. The present study was conducted on the detection of serum antibodies and molecular detection of classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 2 (PCV2) and rotavirus in the pigs. A total of 204 blood and 209 rectal swabs were collected from apparently healthy pigs of Champhai district of Mizoram transported from Myanmar. All the serum samples were subjected to ELISA based detection of antibodies against CSFV, PRRSV and PCV2. The whole blood/serum/feces were processed for extraction of viral nucleic acids and subjected to PCR/RT-PCR for detection of CSFV, PRRSV, PCV2 and rotavirus. A total of 105 (51.47%), 12(5.88%) and 36 (17.65%) serum samples were found to be positive for antibodies against CSFV, PRRSV and PCV2, respectively. In addition, 17 (8.33%) sera were recorded positive for both CSFV and PCV-2 antibodies and 9 (4.41%) sera were positive for CSFV, PCV-2 and PRRSV antibodies. By specific PCR, group A rotavirus was found to be most prevalent (8.33%) followed by PCV-2 (7.84%), CSFV (4.41%) and PRRSV (0.9%). All the rotaviruses were detected as Group A only. Of the 204 animals, 2 (0.98%) pigs showed co-infection with all the four viruses under the study. A total of 5 (2.45%) pigs were positive for co-infection with CSFV and PCV-2 and 4 (1.96%) pigs were positive for co-infection with PCV-2 and rotavirus. The present study provided laboratory based evidence on transmission of porcine viral pathogens from Myanmar to Mizoram.

Serodynamic Analysis of the Piglets Born from Sows Vaccinated with Modified Live Vaccine or E2 Subunit Vaccine for Classical Swine Fever.

Classical swine fever (CSF) caused by the CSF virus (CSFV) is one of the most important swine diseases, resulting in huge economic losses to the pig industry worldwide. Systematic vaccination is one of the most effective strategies for the prevention and control of this disease. Two main CSFV vaccines, the modified live vaccine (MLV) and the subunit E2 vaccine, are recommended. In Taiwan, CSF cases have not been reported since 2006, although systemic vaccination has been practiced for 70 years. Here, we examined the sero-dynamics of the piglets born from sows that received either the CSFV MLV or the E2 vaccine and investigated in the field the correlation between the porcine reproductive and respiratory syndrome virus (PRRSV) loads and levels of CSFV antibody. A total of 1398 serum samples from 42 PRRSV-positive farms were evaluated to determine the PRRSV loads by real-time PCR and to detect CSFV antibody levels by commercial ELISA. Upon comparing the two sow vaccination protocols (CSFV MLV vaccination at 4 weeks post-farrowing versus E2 vaccination at 4–5 weeks pre-farrowing), the lowest levels of CSFV antibody were found in piglets at 5–8 and 9–12 weeks of age for the MLV and E2 groups, respectively. Meanwhile, the appropriate time window for CSFV vaccination of offspring was at 5–8 and 9–12 weeks of age in the MLV and E2 groups, respectively. There was a very highly significant negative correlation between the PRRSV load and the level of CSFV antibody in the CSFV MLV vaccination group (P < 0.0001). The PRRSV detection rate in the pigs from the MLV group (27.78%) was significantly higher than that in pigs from the E2 group (21.32%) (P = 0.011). In addition, there was a significant difference (P = 0.019) in the PRRSV detection rate at 5–8 weeks of age between the MLV (42.15%) and E2 groups (29.79%). Our findings indicate that the vaccination of CSFV MLV in piglets during the PRRSV susceptibility period at 5–8 weeks of age may be overloading the piglet’s immune system and should be a critical concern for industrial pork production in the field.

Identification of E2 with improved secretion and immunogenicity against CSFV in piglets

BackgroundOutbreaks of Classical swine fever virus (CSFV) cause significant economic losses in the swine industry. Vaccination is the major method to prevent and control the disease. As live attenuated vaccines fail to elicit differentiable immunity between infected and vaccinated animals, subunit vaccine was considered as an alternative candidate to prevent and eradicate CSFV. Subunit vaccines present advantages in DIVA immunogenicity and safety. The technology was limited due to the low yield and the high cost with multiple and large doses. The native E2 signal peptide has not been well defined before. Here, the aim of this study is to develop a cost-effective and efficacious E2 vaccine candidate against CSFV with signal peptide and E2 sequence selection.ResultsA novel CSFV E2 sequence (E2ZJ) was identified from an epidemic strain of Zhejiang for outstanding secretion in baculovirus and enhanced immunogenicity. E2 secretion induced with the selected signal peptide, SPZJ (SP23), increase at least 50% as compared to any other signal peptides tested. Besides, unique antigenic features were identified in E2ZJ. As indicated with immunized sera in IFA against CSFV infection, E2ZJ elicited CSFV antibodies at the earlier stage than other E2 types tested in mice. Moreover, higher level of neutralizing and CSFV antibodies against CSFV with E2ZJ was detected than other E2s with the same dosage at 28 dpi. Further, E2ZJ successfully elicited neutralizing immunity in piglets. A single dose of 5 μg of E2ZJ was sufficient to induce protective antibodies against CSFV in piglets and provided 100% protection against lethal virus challenge.ConclusionsOur studies provide evidence that E2ZJ guided by a novel E2 signal peptide (SPZJ) was efficiently secreted and presented significantly improved immunogenicity than conventional E2 vaccines. Moreover, a single dose of 5 μg E2ZJ is efficacious against CSFV in piglets.

E2 and Erns isotype-specific antibody responses in serum and oral fluid after infection with classical swine fever virus (CSFV)

Oral fluid sampling for the detection of classical swine fever virus infection provides a relatively inexpensive method for conducting active CSF surveillance. The purpose of this study was to detect CSFV nucleic acid and antibody in serum and oral fluid samples in a group of 10 pigs infected with the moderate CSFV strain, Paderborn. Based on clinical signs, outcome, and other results, pigs were placed into one of three disease outcome groups; Acute, Chronic and Recovered. Oral fluid and serum samples were analyzed for the presence of CSFV nucleic acid along with E2 and Erns surface protein-specific IgM, IgG and IgA responses. The results were summarized into a timeline of detection events beginning with the appearance of E2-IgM in serum (3 DPI) followed by CSFV nucleic acid in serum (6 DPI), CSFV nucleic acid in oral fluid (8 DPI), E2-IgG in serum (20 DPI), and E2-IgG in oral fluid (24 DPI). The results show that a combination of molecular and serological analyses of oral fluid can be incorporated into CSF surveillance.

Development of Plant-produced E2 Protein for Use as a Green Vaccine Against Classical Swine Fever Virus

Plants are promising host systems for recombinant protein production. However, progress in the commercialization of plant-made proteins (PMPs) has been slow. Only one PMP drug is commercially available. In this study, we explored the possibility of using plants to produce E2 of classical swine fever virus (CSFV) and the use of this plant-produced E2 as a vaccine. We designed high-level expression vectors for transgenic plants by considering the transcription, translation, and storage of E2 in the cell. We incorporated a cellulosebinding domain sequence into the expression vector as an affinity tag for cost-effective, one-step purification. Using this vector, we generated multiple lines of transgenic Arabidopsis thaliana plants expressing a fusion protein of E2 from CSFV at high levels (0.7% of total soluble proteins). ER-targeted E2 fusion protein was successfully purified via a one-step purification process using amorphous cellulose resin. Arabidopsis-produced E2 was recognized by an antibody that detects CSFV antigen. Finally, antisera from mice immunized with E2 fusion protein reacted strongly to the antigens in a CSFV antibody detection kit. Therefore, we propose that plant-produced E2 fusion proteins could be further developed for use as a green vaccine against CSFV in animals.

Standardization and application of rapid tests viz dot ELISA and vertical flow through tests for serodiagnosis of classical swine fever

The present study was carried out to develop rapid field-based immunoassays for onsite detection of Classical swine fever virus (CSFV) antibody in clinical serum sample. Purified CSFV antigen was coated onto nitrocellulose membranes housed in a plastic module with layers of absorbent filter pads underneath. Following addition of serum to be tested appearance of a red dot indicated the presence of antibodies to CSF virus in the sample tested. A total of 240 serum samples collected from different areas of North Eastern States of India were tested for the presence of CSFV antibody by Indirect ELISA, Dot ELISA and Vertical flow through test. The result of the Indirect ELISA could be obtained within 2.5 hours whereas the result in case of Dot ELISA and Vertical Flow through test was obtained within 1 hour and 15 minutes respectively. Dot-ELISA and Vertical flow through test could equally detect the CSFV antibody in samples possessing 1:4 and above ELISA titre. The specificity and sensitivity of Dot-ELISA and vertical flow through test after comparison with Indirect ELISA indicated 100% specific and 84.28% sensitive. These tests have the potential to be used as a field-based kit to assess the serodiagnosis of CSFV.

First-time detection of porcine reproductive and respiratory syndrome virus (PRRSV) infection in Uruguay.

Within the last two decades, several high-impact viruses have emerged in the global swine population, including porcine reproductive and respiratory syndrome virus (PRRSV). In Uruguay, the more recent serological survey for PRRSV and other notifiable diseases such as Aujeszky's disease virus (ADV) and classical swine fever virus (CSFV) dated from year 2000. The main purpose of this study was to update our information on the infection status of PRRSV, ADV and CSFV in Uruguayan pig herds, in order to keep informed about the epidemiological situation of these notifiable infections in the country. For serological testing, a total of 524 swine serum samples collected during the period 2014-2016 were assayed by commercial ELISAs. Our results revealed the (unexpected) presence of PRRSV antibodies in Uruguayan domestic swine herds and confirmed the absence of ADV and CSFV antibodies in all of the assessed samples. Following such initial finding, PRRSV antibodies were further investigated in 23 retrospective samples collected during 2010-2014. Thirteen of these 23 samples resulted seropositive. Subsequently, a molecular detection approach in frozen serum samples was implemented to confirm PRRSV infection, and viral RNA was identified by reverse transcription-nested polymerase chain reaction (RT-nPCR). Fourteen of 86 evaluated 2014-2016 samples resulted positive for viral RNA, while molecular analysis of four retrospective samples also revealed the presence of PRRSV type 2. Viral isolation of selected samples was carried out in porcine alveolar macrophages (PAM) and MARC 145 simian kidney cells, and the virus identity was confirmed by cytopathic effect (CPE) and immunofluorescence assay (IFA) using specific monoclonal antibodies for PRRSV nucleocapsid. Data reported here evidence for the first time the circulation of PRRSV type 2 in Uruguay, and retrospective serology results suggest that the virus has been infecting pigs in this country at least since 2011.

Seroprevalence of selected viral pathogens in pigs reared in organized farms of Meghalaya from 2014 to 16.

Aim:A pilot study was carried out to find out the seroprevalence of Porcine circovirus 2 (PCV2), classical swine fever virus (CSFV), and Porcine respiratory and reproductive syndrome virus (PRRS) in pig population of Meghalaya.Materials and Methods:Serum samples were collected from piglets of 40–45 days age group, growers, and sows reared under organized and unorganized management in 11 districts of Meghalaya situated in the Khasi, Jaintia, and Garo hills divisions in the time period of 2014-2016 from apparently healthy and suspected pigs. Seroprevalence of PCV2, CSFV, and PRRS specific antibodies was detected by enzyme-linked immunosorbent assay (ELISA).Results:A total of 1899 serum samples were collected and screened using antibody ELISA kits specific for PCV2, CSFV, and PRRS. The highest antibody prevalence during the selected time periods was detected for PCV2 (80.8% in 2014, 79.1% in 2015, and 96.2% in 2016) followed by CSFV (76.4% in 2014, 66.09% in 2015, and 25.5% in 2016) and PRRS (2.8% in 2014, 2.7% in 2015, and 3.62% in 2016). The result indicates high seroprevalence for PCV2, which can be considered as an inducement factor due to the immunosuppressive nature of the virus, for animals being susceptible to other pathogens in farms where airborne transmission of PCV2 and postweaning multisystemic wasting syndrome among animals reared in close pens can be a major possibility.Conclusions:The data from this study indicates ubiquitous prevalence of PCV2 antibodies in the farm animals along with the endemic presence of swine fever and emergence of PRRS in an organized farm. There are few reports regarding PCV2 infections/outbreaks in pigs associated with reproductive failure from northern and southern part of India, but till date, there are no reports regarding concomitant infection of CSFV and PCV2 from India. Considerable high seropositivity of PCV2 indicates the need for high impact hygiene practice in farms, routine seromonitoring and implementation the vaccination program. To the author’s best knowledge, this is the first documented report on the seroprevalence of PCV2, CSFV, and PRRS from pig population of Meghalaya.

Prevalence of African swine fever virus and classical swine fever virus antibodies in pigs in Benue State, Nigeria.

This study investigated the prevalence of African swine fever virus (ASFV) and classical swine fever virus (CSFV) antibodies in pigs in Benue State, Nigeria. Serum samples were collected from a total of 460 pigs, including 416 from 74 piggeries and 44 from Makurdi slaughter slab. The samples were analysed using indirect enzyme-linked immunosorbent assay (ELISA) test kit to detect the presence of ASFV antibodies, while competitive ELISA test kit was used to detect antibodies to CSFV. Our findings showed a total ASF prevalence of 13 (2.8%), while prevalences of 7 (1.7%) and 6 (13.6%) were observed in piggeries and in Makurdi slaughter slab, respectively. However, no CSFV antibody sera were detected in this study. Relatively higher ASFV antibody-positive pigs were detected in the slaughter slab than in piggeries. The difference in prevalence of ASF between the two locations was significantly associated (p=0.017). These findings suggest the presence of ASFV antibody-positive pig in Benue State, Nigeria. Continuous surveillance and monitoring of these diseases among pigs in Nigeria to prevent any fulminating outbreak are recommended.

Effects of Vaccination with the C-Strain Vaccine on Immune Cells and Cytokines of Pigs Against Classical Swine Fever Virus.

The attenuated C-strain vaccine against classical swine fever virus (CSFV) is one of the safest and most effective attenuated vaccines. However, little is known of the host immune response after vaccination with the C-strain vaccine. Blood samples from vaccinated pigs were collected to evaluate the number of immune cells, the level of specific CSFV antibody, and related cytokines induced by the vaccination of C-strain vaccine. The C-strain nucleic acid was gradually removed and specific antibody to vaccine kept increasing; the amount of the lymphocyte, Tc cell, and Th cell increased; some inflammatory cytokines such as interleukin (IL)-1 and tumor necrosis factor-α mainly showed downregulated trends, but IL-6 and IL-8 were upregulated greatly; IL-2, IL-4, IL-5, IL-12p40, IL-13, interferon (IFN)-I, and Toll-like receptors (TLRs) kept high expression level after 28 days postvaccination (dpv); IFN-γ was upregulated slightly at 5 and 9 dpv, respectively. These results suggest that the C-strain vaccine induces a Th2 cell response to produce the specific antibody. The vaccine virus replicates at very low level. C-strain vaccine burden has close relationship with the expression of TLRs. The overexpression of TLRs initiates the innate immune system to clear up the vaccine. Meanwhile, ILs expressed by immune system induce the differentiation of B cells and produce specific antibody.

Dietary Supplementation of Astragalus Polysaccharides Enhanced Immune Components and Growth Factors EGF and IGF-1 in Sow Colostrum.

Colostrum is the main external resource providing piglets with nutrients and maternal immune molecules. Astragalus polysaccharides (APS) have been used as immunopotentiators in vitro and several animal models. This study aimed to determine the effects of APS on immune factors in sow colostrum and milk. The sow diet was supplemented with APS one week before the expected delivery date. Colostrum and milk were collected and designated as 0 h- (onset of parturition), 12 h-, and 24 h-colostrum and 36 h-milk postpartum. Samples were measured using porcine immunoglobulin (Ig) G, IgM, classical swine fever virus antibody (CSFV Ab), epidermal growth factor (EGF), and insulin-like growth factor- (IGF-) 1 ELISA Quantitation Kits. Dietary supplementation of APS significantly enhanced the presence of IgG, IgM, EGF, and IGF-1 in 0 h-colostrum (P < 0.001). The blocking rates of CSFV Ab were increased in samples from APS-supplemented sow when compared to those from the matched samples without APS treatment. The results indicate that supplement of APS could improve the immune components in sow colostrum and/or milk; and status of some specific vaccination could be determined through using colostrum or early milk in sow.

Impact of porcine reproductive and respiratory syndrome virus and porcine circovirus-2 infection on the potency of the classical swine fever vaccine (LOM strain)

The classical swine fever (CSF) vaccine, which is derived from the LOM strain of the CSF virus (CSFV), induces protective immunity against CSFV infection. However, several factors influence vaccine efficacy. Evidence suggests that infection by porcine reproductive and respiratory syndrome virus (PRRSV) and/or porcine circovirus 2 (PCV2) reduces the efficacy of several vaccines. Here, we examined the effect of PRRSV or PCV2 alone or co-infection by PRRSV/PCV2 on the potency of the LOM vaccine in pigs. Neither CSFV antibody levels nor the period during which CSFV antigens were detectable in LOM-vaccinated pigs were negatively affected by infection by PRRSV or PCV2. However, co-infection with PRRSV/PCV2 may affect the replication or activity of the CSF vaccine virus in pigs vaccinated with the LOM strain, although CSFV antibody levels were not negatively affected. Nevertheless, the LOM vaccine afforded complete protection against a virulent strain of CSFV.

Development of single dilution immunoassay to detect E2 protein specific classical swine fever virus antibody

Classical swine fever virus (CSFV) is the causative agent of a highly contagious disease in swine. The disease is endemic in different parts of the world and vaccination is the only way to protect pigs from CSFV infection. The virus surface protein E2 is the major immunogenic protein eliciting protective immunity against CSFV infection in swine. The whole virus antigen cannot differentiate CSFV from other pestiviruses as it cross reacts with border disease and bovine viral diarrhoea viruses. Commercial available ELISA is based on the whole CSFV particle and can lead to false positive results. Moreover, the available commercial ELISA is not cost effective. In the present study, a recombinant E2 protein based single serum dilution ELISA was developed which showed enhanced sensitivity, specificity and accuracy as compared to commercial CSFV detection ELISA. The recombinant E2 protein based ELISA could be an alternate to existing diagnostics against CSFV infection in pigs.

Safety of classical swine fever virus vaccine strain LOM in pregnant sows and their offspring

The present study aimed to evaluate the safety of the classical swine fever virus (CSFV) vaccine strain LOM in pregnant sows. Pregnant sows with free CSFV antibody were inoculated with a commercial LOM vaccine during early pregnancy (day 38; n=3) or mid-pregnancy (days 49–59; n=11). In pregnant sows vaccinated during the early stages of gestation, abortion (day 109) was observed in one case, with two stillbirths and seven mummified fetuses. The viability of live-born piglets was 34.9% in sows vaccinated during mid-pregnancy compared with 81.8% in the control group. Post-mortem examination of the organs of the sows and piglets did not reveal any pathological lesions caused by CSFV; however, CSFV RNA was detected in the organs of several vaccinated sows and their litters. The LOM strain was transmitted from sows with free CSFV antibody to their fetus, but did not appear to induce immune tolerance in the offspring from vaccinated pregnant sows. Side effects were not observed in pregnant sows with antibody to the LOM strain: transmission from sow to their litters and stillbirth or mummified fetuses. The LOM strain may induce sterile immunity and provide rapid, long-lasting, and complete protection against CSFV; however, it should be contraindicated in pregnant sows due to potential adverse effects in pregnant sows with free CSFV antibody.