Proliferative enteropathy (PE; ileitis) is a common intestinal disease affecting susceptible pigs raised under various management systems around the world. Major developments in the understanding of PE and its causative agent, Lawsonia intracellularis, have occurred that have led to advances in the detection of this disease and methods to control and prevent it. Diagnostic tools that have improved overall detection and early onset of PE in pigs include various serological and molecular-based assays. Histological tests such as immunohistochemistry continue to be the gold standard in confirming Lawsonia-specific lesions in pigs post mortem. Despite extreme difficulties in isolating L. intracellularis, innovations in the cultivation and the development of pure culture challenge models, have opened doors to better characterization of the pathogenesis of PE through in vivo and in vitro L. intracellularis-host interactions. Advancements in molecular research such as the genetic sequencing of the entire Lawsonia genome have provided ways to identify various immunogens, metabolic pathways and methods for understanding the epidemiology of this organism. AbstractProliferative enteropathy (PE; ileitis) is a common intestinal disease affecting susceptible pigs raised under various management systems around the world. Major developments in the understanding of PE and its causative agent, Lawsonia intracellularis, have occurred that have led to advances in the detection of this disease and methods to control and prevent it. Diagnostic tools that have improved overall detection and early onset of PE in pigs include various serological and molecular-based assays. Histological tests such as immunohistochemistry continue to be the gold standard in confirming Lawsonia-specific lesions in pigs post mortem. Despite extreme difficulties in isolating L. intracellularis, innovations in the cultivation and the development of pure culture challenge models, have opened doors to better characterization of the pathogenesis of PE through in vivo and in vitro L. intracellularis-host interactions. Advancements in molecular research such as the genetic sequencing of the entire Lawsonia genome have provided ways to identify various immunogens, metabolic pathways and methods for understanding the epidemiology of this organism. The determinations of immunological responsiveness in pigs to virulent and attenuated isolates of L. intracellularis and identification of various immunogens have led to progress in vaccine development.

Oral administration by drench or via drinking water of an avirulent live vaccine against L. intracellularis resulted in substantial protection against proliferative enteropathy among vaccinates and offers a better way to reduce stress of pigs during vaccine administration.

Adjuvants enhance both the magnitude and duration of immune responses, therefore representing a central component of vaccines. The nature of the adjuvant can determine the particular type of immune response, which may be skewed toward cytotoxic T cell (CTL) responses, antibody responses, or particular classes of T helper (Th) responses and antibody isotypes. Traditionally, adjuvants have been added to intrinsically poor immunogenic vaccines, such as those using whole killed organisms or subunit vaccines. Here, we have compared cellular immune responses induced by the immunogenic modified life-attenuated vaccine Ingelvac PRRS® MLV when administered alone or in combination with carbopol, a widely used adjuvant in veterinary medicine. Using functional readouts (IFN-γ ELISpot and cell proliferation) and analyzing phenotypical hallmarks of CD4T cell differentiation, we show that carbopol improves cellular immunity by inducing early IFN-γ-producing cells and by preferentially driving T cell differentiation to effector phenotypes. Our data suggest that adjuvants may enhance and modulate life-attenuated--not only subunit/inactivated--vaccines.

An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived fromProliferative enteropathy (PE) is a common enteric disease of pigs after weaning that is caused by the obligate intracellular bacterium, Lawsonia intracellularis (20, 21). The infection and disease are widespread in pig farms across America and Europe, and prevalence among groups of pigs on these affected farms can be over 30% (4,23,29,31). This leads to a considerable economic impact of the disease, due to diarrhea, weight loss, and subclinical illness in growing pigs (22,31). Since the identification of L. intracellularis as the cause of PE in 1993, a number of studies aimed at establishing the best diagnostic methods for identifying Lawsonia exposure in live animals have been conducted. These have focused on DNA detection via PCR of feces and whole-cell immunoassays (8,12,13,15,17), due to the extreme difficulty of isolation of the obligate intracellular L. intracellularis from the contaminated environment of feces (13,17,18). In situations where samples of ileum are available, immunohistochemistry (IHC) is considered to provide the criterion-referenced measure or "gold standard" for assessment of the actual infection status of an individual pig (9,16,19,26,28).PCR testing of fresh feces involves considerable laboratory effort and cost to extract amplifiable bacterial DNA from each sample (9,11,13,15). False positives due to pre-laboratory sample contamination during the collection of numerous samples from a group of pigs or due to contamination during the laboratory testing phase may occur. False negatives due to the regular presence of PCR inhibitors in feces may also occur (9, 10, 11). Serologic testing methods have therefore also been widely explored for detecting L. intracellularis exposure of pigs. Indirect immunofluorescence or immunoperoxidase assays have been used to examine antibody responses of pigs infected experimentally with L. intracellularis in virulent challenge exposure studies and of pigs with PE from farms (3,4,7,11,14,29). An indirect enzyme-linked immunosorbent assay (ELISA) was developed previously for testing pig serum antibodies, with crude antigen derived directly from pig intestines affected with PE (12). However, the antigen used in that study was not fully characterized for L. intracellularis content. The development of a specific L. intracellularis antigen-based ELISA would therefore be of considerable benefit in improving the feasibility of a more universally available and standardized diagnostic assay to study the epidemiology of this economically significant disease.We describe the development of an ELISA for detecting L. intracellularis infection based on a lipopolysaccharide antigen extract in an indirect ELISA format. MATERIALS AND METHODSBacterial antigen preparation. The lipopolysaccharide (LPS) used in this study was derived from L. intr...

BackgroundPorcine reproductive and respiratory syndrome (PRRS) causes respiratory distress in pigs, reproductive failure in breeding-age gilts and sows, and can have devastating economic consequences in domestic herds. Several PRRS vaccines are available commercially. This study compared the effectiveness of single-vaccination and revaccination schedules using the PRRS 94881 Type I modified live virus (MLV) vaccine ReproCyc® PRRS EU with no vaccination (challenge control) in protecting against a PRRS virus (PRRSV) challenge in non-pregnant gilts.ResultsData were available from 48 gilts across three groups: a challenge control group (n = 16), which received no vaccination; a revaccination group (n = 16), which received ReproCyc® PRRS EU on Days 0 and 56; and a single vaccination group (n = 16), which received ReproCyc® PRRS EU on Day 56. All gilts were PRRSV RNA-negative (based on reverse transcription and quantitative polymerase chain reaction [RT-qPCR]) and PRRSV seronegative (based on enzyme-linked immunosorbent assay [ELISA]) at Day 0. All gilts were challenged with PRRSV strain 190136 on Day 91.Viral RNA loads in both vaccination groups were significantly reduced compared with the challenge control group on Days 98 (P < 0.0001) and 101 (P < 0.0001), indicating that vaccinated gilts were better able to respond to challenge than unvaccinated gilts. At all timepoints following challenge, mean viral RNA load and the percentage of PRRSV RNA-positive gilts were numerically higher in the single-vaccination group than in the revaccination group; these differences were statistically significant on Day 101 (P = 0.0434). Furthermore, viremia levels after challenge were significantly lower in the revaccination group than in the single-vaccination group based on median area under the curve (AUC) values for viral RNA load from Day 91 to Day 112, suggesting that revaccinated gilts had better protection from viral infection than gilts who received a single vaccination. Protection from viremia did not correlate with the proportion of seropositive gilts on Day 91. In the single-vaccination group, 94% of pigs were seropositive on Day 91 compared with 56% in the revaccination group. Vaccination was well tolerated and no safety concerns were identified.ConclusionsBoth single-vaccination and revaccination with ReproCyc® PRRS EU were effective in reducing PRRSV viremia post-challenge. These findings have important implications for herd management as both the single-vaccination and revaccination schedules protect against PRRSV challenge, with revaccination appearing to provide better protection from viremia than single vaccination.

BackgroundPRRS is a viral disease of pigs and sows that is one of the most costly to the pig industry worldwide. The disease can be controlled by focusing on different aspects. One of them is the vaccination of piglets, which is more controversial and difficult to manage than the vaccination of sows. However, pig producers could consider a piglet vaccination strategy if it reduces the negative clinical disease and improves zootechnical performance, decreases the probability to be infected and/or reduces the spread of the virus once the vaccinated piglet is infected. The efficacy of a novel PRRS modified live vaccine (Ingelvac PRRSFLEX® EU) was studied in a blinded, side-by-side placebo controlled field study of piglet vaccination including piglets weaned for three consecutive weeks (week groups 1, 2 and 3).ResultsThis study established that PRRS piglet vaccination resulted in significantly better weight gain, seen as early as 4 weeks after vaccination, in naturally challenged pigs. Vaccine efficacy was supported by statistically significant increases in Average Daily Weight Gain (ADWG) among week group 3 vaccinated pigs from vaccination to the end of the study and statistically significant increases in bodyweight and ADWG from inclusion to 10 weeks of age in week group 2 vaccinated piglets. However, no differences were noted in week group 1 presumably because more than 30 % of the vaccinated pigs were viremic at the time of vaccination. Furthermore, the proportion of pigs showing any abnormal clinical sign at least once at any of the examination time points was lower in vaccinated pigs than in control pigs. Based on the viremia results (qPCR), early onset of PRRS was detected in this herd. Viremia occurred at the time of vaccination in week group 1 and shortly after vaccination in week groups 2 and 3. Peak wild type PRRSV infection was assumed at 4 weeks post vaccination in all groups based on the number of PRRS positive pigs in the control groups.ConclusionThis study establishes that vaccination of piglets with Ingelvac PRRSFLEX® EU at 4 weeks of age improves weight gain and reduces the appearance of clinical sings during the growing period, even when the piglets are infected shortly after vaccination.Electronic supplementary materialThe online version of this article (doi:10.1186/s40813-016-0038-x) contains supplementary material, which is available to authorized users.

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Abstract. In the current study, the development and validation of a real-time polymerase chain reaction (PCR) assay using a TaqMan-labeled probe for the detection of Actinobacillus suis from porcine lung samples is described. This real-time PCR amplified a 110-bp region of the 23S ribosomal RNA gene from A. suis but not from other bacteria. First, the assay was validated with 183 bacterial strains representing different species of bacteria. Subsequently, 85 porcine lung specimens that were declared A. suis-positive and -negative by bacterial culture and identification were tested to assess whether it can be performed directly on tissue specimens. The bacterial culture results and real-time PCR results agreed across all the samples tested assigning 100% positive and negative predictive values to the PCR. Further, the detection limit of the assay was 380 colony-forming units (CFU) per ml or approximately 1 CFU per reaction. In conclusion, the TaqMan real-time PCR assay described herein is a highly specific, sensitive, and reproducible test, which can be used to detect A. suis DNA in porcine lung specimens, thus providing a timely diagnosis.