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Highlights • Delivery of G-CSF to pigs via an adenovirus vector induced a sustained increase in circulating neutrophil numbers. • There was a longer mean survival time after Streptococcus suis challenge in pigs treated with G-CSF. • G-CSF may be a useful adjunct to antibiotics for prevention of Streptococcal disease in swine. Abstract The use of immunomodulators is a promising alternative to the use of antibiotics for therapeutic, prophylactic, and metaphylactic use to prevent and combat infectious disease. Previously we demonstrated a replication-defective adenovirus vector that expresses porcine granulocyte colony-stimulating factor (G-CSF) elicited a sustained neutrophilia, lasting nearly 3 weeks, which may be beneficial to prevent bacterial diseases during times of peak incidence. In a pilot study using the vectored G-CSF with a Caesarian-derived, colostrum-deprived (CDCD) pig model of Streptococcus suis disease, only 1 of 4 pigs given G-CSF developed disease, while 3 of 4 non-treated pigs developed Streptococcal disease. In a subsequent study using a larger number of pigs, although there was no difference in overall survival, there was a longer mean survival time in G-CSF treated pigs. S. suis infection is more severe in CDCD pigs than conventionally raised pigs, consequently results in the field may be superior to the ones reported in this study. Although there were positive effects from the use of G-CSF in this study, further research is needed to determine if improved clinical outcomes could be achieved under field conditions and whether the use of G-CSF in pigs to induce a sustained increase in circulating neutrophil numbers may be useful as an adjunct to antibiotics to diminish the severity of Streptococcal disease, especially during times of stress and pathogen exposure such as post-weaning. [ABSTRACT FROM AUTHOR]

Abstract Understanding between-farm movement patterns is an essential component in developing effective surveillance and control programs in livestock populations. Quantitative knowledge on movement patterns is particularly important for the commercial swine industry, in which large numbers of pigs are frequently moved between farms. Here, we described the annual movement patterns between swine farms in three production systems of the United States and identified farms that may be targeted to increase the efficacy of infectious disease control strategies. Research results revealed a high amount of variability in movement patterns across production systems, indicating that quantities captured from one production system and applied to another may lead to invalid estimations of disease spread. Furthermore, we showed that targeting farms based on their mean infection potential, a metric that captured the temporal sequence of movements, substantially reduced the potential for transmission of an infectious pathogen in the contact network and performed consistently well across production systems. Specifically, we found that by targeting farms based on their mean infection potential, we could reduce the potential spread of an infectious pathogen by 80% when removing approximately 25% of farms in each of the production systems. Whereas other metrics, such as degree, required 26–35% of farms to be removed in two of the production systems to reach the same outcome; this outcome was not achievable in one of the production systems. Our results demonstrate the importance of fine-scale temporal movement data and the need for in-depth understanding of the contact structure in developing more efficient disease surveillance and response strategies in swine production systems. [ABSTRACT FROM AUTHOR]

Porcine Reproductive and Respiratory Syndrome (PRRS) is an elusive model of host/virus relationship in which disease is determined by virus pathogenicity, pig breed susceptibility and phenotype, microbial infectious pressure and environmental conditions. Successful disease control in PRRS-endemic Countries corresponds to “stability”, i.e. a condition with no clinical signs of PRRS in the breeding-herd population and no viremia in weaning-age pigs. The aim of this work was to compare the profile and time-course of humoral and cell-mediated immunity in stable and unstable herds, respectively. In particular, we investigated PRRS virus (PRRSV) in serum and group oral fluid samples by Real-time RT-PCR, PRRSV-specific IgA and IgG in oral fluids, serum IgG antibody and the cell-mediated response (PRRSV-specific release of interferon-gamma) in whole blood samples. These parameters were measured in order to identify possible discrepancies in the development and kinetics of the immune response against PRRSV. PRRS-free gilts got regularly infected after entering PRRS-stable and unstable farms. In an open cycle, unstable pig farm PRRSV infection could be demonstrated in all groups of pigs, including suckling piglets. Four main results should be highlighted: A) the precocity of the Ab response in group oral fluids was generally similar to that recorded in sera; B) circulation of PRRSV was consistently detected in all age groups in the unstable herds, as opposed to the stable ones; C) an early, balanced, IgA and IgG response in oral fluids was only observed in the stable herds; D) an early IFN-gamma response after PRRSV infection was often observed in stable herds, as opposed to the unstable ones. These were characterized by IFN-gamma responses in piglets, likely due to transfer of maternal immunity. Most important, the mucosal IgA response was associated with cessation of virus excretion in oral fluid samples of PRRS-unstable herds. The above findings indicate that a peculiar profile of immune response to PRRSV can be found in PRRS-stable herds. Therefore, the outlined immune parameters can represent a useful readout system to evaluate successful adaptation to PRRSV based on acclimatization of breeding animals and management of pig flow. [ABSTRACT FROM AUTHOR]

The nucleocapsid (N) protein is the most abundant protein of porcine reproductive and respiratory syndrome virus (PRRSV). It has been shown to be multiphosphorylated. However, the phosphorylation sites are still unknown. In this study, we used liquid chromatography tandem mass spectrometry (LC–MS/MS) to analyze the phosphorylation sites of N protein expressed in Sf9 cells. The results showed that N protein contains two phosphorylation sites. Since N protein can regulate IL-10, which may facilitate PRRSV replication, we constructed four plasmids (pCA-XH-GD, pCA-A105, pCA-A120 and pCA-A105-120) and transfected them into Pig alveolar macrophages (PAMs,3D4/2). The qPCR results showed that mutations at residues 105 and 120 were associated with down-regulation of the IL-10 mRNA level, potentially decreasing the viral growth ability. Then, we mutated the phosphorylation sites (S105A and S120A) and rescued three mutated viruses, namely, A105, A120 and A105-120. Compared with wild-type virus titers, the titers of the mutated viruses at 48 hpi were significantly decreased. The Nsp(non-structural protein) 9 qPCR results were consistent with the multistep growth kinetics results. The infected PAMs (primary PAMs) results were similar with Marc-145.The findings indicated that the mutations impaired the viral replication ability. The confocal microscopy results suggested that mutations to residues 105 and 120 did not affect N protein distribution. Whether the mutations affected other functions of N protein and what the underlying mechanisms are need further investigation. In conclusion, our results show that residues 105 and 120 are phosphorylation sites and are important for N protein function and for viral replication ability. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS) and classical swine fever (CSF) are economically significant diseases that affect the swine industry worldwide. However, the current vaccination strategy, which uses two single live attenuated vaccines, can result in interference for each other. In addition, the universally used CSFV vaccine C-strain does not allow for differentiation of infected and vaccinated animals. In this study, rPRRSV-E2, PRRS virus (PRRSV) expressing CSF virus (CSFV) E2, was constructed by reverse genetics. The E2 gene of CSFV was inserted between ORF1b and ORF2 in the genome of the PRRS vaccine virus, HuN4-F112. A copy of transcriptional regulatory sequence 6 was inserted at the 3′ terminal of the exogenous gene to produce CSFV E2 as a unique subgenomic mRNA transcript. The rPRRSV-E2 was stable for at least 25 serial cell passages. Single-shot intramuscular immunization of rPRRSV-E2 into pigs induced PRRSV-specific and CSFV-specific antibodies and fully protected pigs from lethal challenge with highly-pathogenic PRRSV and CSFV. These results demonstrate that a novel strategy for recombinant PRRSV production is effective, and suggest that rPRRSV-E2 is a promising live, virus-vectored vaccine against PRRS and a marker vaccine against CSF. [ABSTRACT FROM AUTHOR]

Modified-live virus (MLV) vaccines are widely used to protect pigs against porcine reproductive and respiratory syndrome virus (PRRSV). However, current MLV vaccines do not confer adequate levels of heterologous protection, presumably due to the substantial genetic diversity of PRRSV isolates circulating in the field. To overcome this genetic variation challenge, we recently generated a synthetic PRRSV strain containing a consensus genomic sequence of PRRSV-2. We demonstrated that our synthetic PRRSV strain confers unprecedented levels of heterologous protection. However, the synthetic PRRSV strain at passage 1 (hereafter designated CON-P1) is highly virulent and therefore, is not suitable to be used as a vaccine in pigs. In the present study, we attenuated CON-P1 by continuously passaging the virus in MARC-145 cells, a non-natural host cell line. Using a young pig model, we demonstrated that the synthetic virus at passages 90 and 122 (designated as CON-P90 and CON-P122, respectively) were fully attenuated, as evidenced by the significantly reduced viral loads in serum and tissues and the absence of lung lesion in the infected pigs. Most importantly, CON-P90 confers similar levels of heterologous protection as its parental strain CON-P1. Taken together, the results indicate that CON-P90 is an excellent candidate for the formulation of next generation of PRRSV MLV vaccines with improved levels of heterologous protection. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSV), a single-stranded, positive-sense RNA virus of the Arteriviridae family, has become a global health threat for swine. Cholesterol 25-hydroxylase (CH25H) is an enzyme that catalyzes oxidation of cholesterol to 25-hydroxycholesterol (25HC). The purpose of this study was to explore the antiviral activity of CH25H against PRRSV infection. We found that CH25H was induced by interferon-α and PRRSV in Marc-145 monkey kidney cells. In addition, CH25H and 25HC significantly inhibited PRRSV infection by preventing virus entry. A CH25H mutant that exhibited decreased catalytic activity had an antiviral effect against PRRSV. Treatment with 25HC pre-infection or post-infection significantly inhibited PRRSV infection in primary porcine alveolar macrophages. Our results reveal that CH25H is an interferon-stimulated gene and its production of 25HC can be used as a natural antiviral agent to combat PRRSV infection. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSv) is an economically significant swine pathogen causing production losses in the global swine industry. Clinical impact depends on many factors including the virus itself. One method to sub-type PRRSv is using restriction fragment length polymorphism (RFLP). The RFLP pattern 1-7-4 emerged within the United States swine industry in 2014 and has become prevalent since then. This was a field study that prospectively followed 1-7-4-infected breeding herds (n = 107) and compared time to stability (TTS), time to baseline production (TTBP) and total loss per 1000 sows between herds using modified-live virus vaccine (MLV) on sows and gilts (MLV-MLV), MLV on sows and MLV in addition to field virus exposure on gilts (MLV–MLV/FVE) or not deliberately exposing sows or gilts to PRRSv (Natural-Natural). Analyses were done in SAS 9.4 and results were adjusted by selected co-variates (duration of herd closure, number of previous PRRSv outbreaks of last 3 years, weaning frequency/week, gilt development unit location, herd size and production system). Survival analysis was conducted on TTS and TTBP and regression analysis on total loss. Herds in the Natural–Natural group achieved TTS and TTBP before other herds. Herds in the MLV-MLV/FVE had the longest TTS and TTBP. The total loss was numerically least in MLV–MLV herds (1194 pigs/1000 sows) compared to MLV/MLV-FVE (1810/1000 sows) and Natural–Natural (2671/1000 sows). This study provided additional information to assist veterinarians deciding between methods of exposure to manage PRRSv infection from breeding herds. [ABSTRACT FROM AUTHOR]

The recent introduction of the porcine epidemic diarrhea virus (PEDV) into the North American swine herd has highlighted again the need for effective vaccines for swine coronaviruses. While vaccines for transmissible gastroenteritis virus (TGEV) have been available to producers around the world for a long time, effective vaccines for PEDV and deltacoronaviruses were only recently developed or are still in development. Here, we review existing vaccine technologies for swine coronaviruses and highlight promising technologies which may help to control these important viruses in the future. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to afflict swine nearly 30 years after it was first discovered as the causative agent of “mystery swine disease”. Immunological tools of vaccination and exposure to virulent viruses have not succeeded in achieving control and prevention of PRRSV. Humoral immunity, mediated by antibodies, is a hallmark of anti-viral immunity, but little is known about the effector mechanisms of humoral immunity against PRRSV. It is essential to understand the immunological significance of antibody functions, including recently described broadly neutralizing antibodies and potential non-neutralizing activities, in the immune response to PRRSV. Here, we review recent research from PRRSV and other host-pathogen interactions to inform novel routes of exploration into PRRSV humoral immunity which may be important for identifying the immunological correlates of protection against PRRSV infection. [ABSTRACT FROM AUTHOR]

Salmonella carriage in pigs is a significant food safety issue. Dietary supplementation with organic acids has previously been shown to reduce shedding and transmission of Salmonella . Therefore, this study aimed to examine the effect of three commercially available organic acid-based products on Salmonella levels in grower pigs, using a model of experimental infection that closely mimics natural exposure to the organism. Seven week old trial pigs (n = 40) with a mean weight of 14.7 kg were placed in one of four pens with 10 pigs/pen. Pens had previously been contaminated with Salmonella Typhimurium 4,[5],12;i;- via seeder pigs. Trial pigs received one of four diets for 28 days: 1, control diet; 2, sodium butyrate supplemented diet; 3, benzoic acid supplemented diet and 4, formic-citric acid supplemented diet. A further 10 pigs were placed in a Salmonella -free pen receiving the control diet. Pigs were weighed and blood sampled on days 0 and 28. Faeces was collected on day 0, 2, 3, 5, 7, 14, 21 and 28 and examined for Salmonella . On day 28, 5 pigs/group were euthanised and ileocaecal lymph nodes (ILN) and caecal contents sampled for culture. The remaining 5 pigs/pen were then fed the control diet and faeces were collected on days 35 and 42. On day 42 pigs were euthanised and ILN and caecal contents tested for Salmonella levels. The trial was repeated once. Within the first two days of exposure to the contaminated environment, 96% (77/80) of pigs became infected. Most pigs shed Salmonella at levels of between 10 0 –10 3 CFU/g faeces for at least 7 days post-exposure. A significant reduction in Salmonella faecal concentration was observed after supplementation with sodium butyrate (p = 0.001) and a formic citric acid blend (p < 0.0001). Average daily weight gain (ADWG) was significantly increased in all groups fed the supplemented feed when compared to the positive control group. The use of sodium butyrate or a blend of formic and citric acid in feed could be considered a cost-effective control measure to reduce Salmonella faecal shedding and improve ADWG in Salmonella infected herds. [ABSTRACT FROM AUTHOR]

Current porcine reproductive and respiratory syndrome virus (PRRSV) vaccines sometimes fail to provide adequate immunity to protect pigs from PRRSV-induced disease. This may be due to antigenic differences among PRRSV strains. Rapid production of attenuated farm-specific homologous vaccines is a feasible alternative to commercial vaccines. In this study, attenuation and efficacy of a codon-pair de-optimized candidate vaccine generated by synthetic attenuated virus engineering approach (SAVE5) were tested in a conventional growing pig model. Forty pigs were vaccinated intranasally or intramuscularly with SAVE5 at day 0 (D0). The remaining 28 pigs were sham-vaccinated with saline. At D42, 30 vaccinated and 19 sham-vaccinated pigs were challenged with the homologous PRRSV strain VR2385. The experiment was terminated at D54. The SAVE5 virus was effectively attenuated as evidenced by a low magnitude of SAVE5 viremia for 1–5 consecutive weeks in 35.9% (14/39) of the vaccinated pigs, lack of detectable nasal SAVE5 shedding and failure to transmit the vaccine virus from pig to pig. By D42, all vaccinated pigs with detectable SAVE5 viremia also had detectable anti-PRRSV IgG. Anti-IgG positive vaccinated pigs were protected from subsequent VR2385 challenge as evidenced by lack of VR2385 viremia and nasal shedding, significantly reduced macroscopic and microscopic lung lesions and significantly reduced amount of PRRSV antigen in lungs compared to the non-vaccinated VR2385-challenged positive control pigs. The nasal vaccination route appeared to be more effective in inducing protective immunity in a larger number of pigs compared to the intramuscular route. Vaccinated pigs without detectable SAVE5 viremia did not seroconvert and were fully susceptible to VR2385 challenge. Under the study conditions, the SAVE approach was successful in attenuating PRRSV strain VR2385 and protected against homologous virus challenge. Virus dosage likely needs to be adjusted to induce replication and protection in a higher percentage of vaccinated pigs. [ABSTRACT FROM AUTHOR]

Strains of Leptospira interrogans belonging to two very closely related serovars – Bratislava and Muenchen – have been associated with disease in domestic animals, in particular pigs, but also in horses and dogs. Similar strains have also been recovered from various wildlife species. Their epidemiology is poorly understood. Two hundred and forty seven such isolates, from UK domestic animal and wildlife species, were examined by restriction endonuclease analysis in an attempt to elucidate their epidemiology. A representative sub-sample of 65 of these isolates was further examined by multiple-locus variable-number tandem repeat analysis and 22 by secY sequencing. Ten restriction pattern types were identified. The majority of isolates fell into one of three restriction endonuclease analysis pattern types designated B2a, B2b and M2a. B2a was ubiquitous and was isolated from 10 species and represented the majority of the horse and all dog isolates. B2b was very different, being isolated only from pigs, indicating that this type was maintained by pigs. The pattern M2a was reported for the majority of isolates from pigs but also was common in small rodents isolates. Five restriction pattern types were found only in wildlife suggesting that they are unlikely to pose a disease threat to domestic animals. Multiple-locus variable-number tandem repeat analysis identified six clusters. The REA types B2a and B2b were all found in one MLVA cluster while the majority of the M2a strains examined occurred in another cluster. The secY sequencing detected only one sequence type, clustered with other serovars of Leptospira interrogans. [ABSTRACT FROM AUTHOR]

Parturation is an intrinsically risky and painful process for both the sow and the piglets that can cause welfare and economic problems. Non-steroidal anti-inflammatory drugs (NSAIDs) have been demonstrated to partially alleviate inflammation and pain after farrowing in sows. NSAIDs effects on piglet mortality and performance show discrepancies and no previous studies have investigated the underlying mechanism. The effects of oral meloxicam treatment to sows on immunoglobulin G (IgG) transfer to piglets around farrowing were investigated. A total of 30 multiparous sows were randomly treated with either oral meloxicam or a mock administration as control group. Treatment was administered as soon as possible at the beginning of the farrowing. A total of 325 piglets were individually weighed at farrowing (day 0) and at weaning (day +21) and piglet mortality was registered during lactation. Four piglets per sow (two piglets suckling from anterior teats and two piglets suckling from posterior teats) were selected for blood sampling at day +1, day +2 and day +20 for IgG analyses. Oral meloxicam treatment to sows significantly increased weight at weaning (mean ± SE: 6563 ± 86.3 g from oral meloxicam group and 6145 ± 103.2 g from control group; P = 0.0017) and ADG (mean ± SE: 236 ± 3.4 g/day from oral meloxicam group and 217 ± 4.5 g/day from control group; P < 0.001) during lactation, but failed to reduce piglet mortality during lactation (6.7% from oral meloxicam group and 6.8% from control group; P = 0.89). IgG levels in piglets from the sows treated with oral meloxicam were significantly higher than the control group at day +1 (mean; median [95% CI] for median = 31.9; 31.7 [29.6–33.6] vs. 27.9; 26.8 [25.9–28.3] mg/ml, P = 0.0013) and day +2 (27.6; 27.0 [24.8–29.6] vs. 24.5; 24.2 [22.1–25.3] mg/ml, P = 0.01). However, at day +20, IgG level in piglet serum was not significantly affected by the treatment (7.6; 7.6 [6.7–8.4] vs. 7.1; 6.9 [6.4–7.3] mg/ml, P = 0.59). The administration of meloxicam orally at the beginning of the farrowing in multiparous sows increased the concentration of IgG in serum of piglets and enhanced their pre-weaning growth. Future research is warranted to clearly identify the proximate mechanism behind IgG effect. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS) virus causes severe and persistent disease in pigs worldwide. Its heterogeneity poses a major challenge for the effective prevention and control of PRRS. Purified viruses are essential for serological studies. Traditional methods for purifying PRRS virus are time- and labor-intensive and difficult to scale-up and requires long processing time. Here, we describe a rapid, simple, scalable process for PRRS virus purification. Highly pure viral particles were obtained after ultrafiltration and liquid chromatography, as confirmed by SDS-PAGE and electron microscopy. The overall process achieved a recovery of 50% of raw virus, with a purity close to that obtained by CsCl coupled with sucrose density gradients. The purification process described here should be useful in large-scale production of highly pure PRRS virus. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) is the etiologic agent of PRRS, one of the most important diseases in swine worldwide. In the present work, the effects of different PRRSV strains were tested on a piglet experimental model to study the induced acute phase response. For this purpose, pigs ( n = 15 for each group) were intranasally inoculated with one of five PRRSV strains (isolates EU10, 12, 17, 18 from genotype 1 and isolate JA-142 from genotype 2). The acute phase response was monitored by measuring acute phase proteins (APPs). Specifically, the serum concentration of haptoglobin (Hp), C-reactive protein (CRP) and Pig-Major Acute Protein (Pig-MAP) was determined at 0, 3, 6, 9, 12, 15, 18 and 21 days p.i. Clinical signs and growth performance were also monitored during the experiment. All animals became viremic after inoculation during the study period. The APP response was heterogeneous and dependent on the strain, being strains EU10, EU 18 and JA-142 those that induced the highest response and the strongest clinical signs. In general, Hp was the most sensitive biomarker for PRRSV infection, CRP behaved as moderate and Pig-MAP was the less responsive during the course of PRRSV experimental infection. Hp and CRP were significantly discriminatory between infected and control pigs, but not Pig-MAP. [ABSTRACT FROM AUTHOR]

Knowledge about the dynamics of methicillin-resistant Staphylococcus aureus (MRSA) in pigs lacks detail at the level of individual animal. The aim of our study was therefore to determine the colonisation status of MRSA in individual pigs from birth to slaughter in order to gain a better understanding of substantial factors involved in transmission. Two farrow-to-finish and two grow-to-finish herds were included in the study. A total of 1728 nasal swabs from 390 pigs and 592 environmental wipes were collected at 11 different time points. Intermittent colonisation throughout the entire production cycle was conspicuous in the tracking of MRSA in individual pigs. Almost all pigs from a MRSA-positive herd changed MRSA status several times, which implies that pigs are transiently rather than permanently colonised. We highly recommend the definition of MRSA status at herd level rather that at the level of the individual pig when considering prevention measures against MRSA. Therefore, to avoid the further spread of MRSA in countries with moderate prevalence, such as in Switzerland, defining farms as MRSA positive or MRSA negative and allowing the trade of pigs only within herds of the same status seems feasible. This will also be important for combating the further dissemination of livestock-associated (LA)-MRSA into healthcare facilities and the community via humans who have close contact with animals. [ABSTRACT FROM AUTHOR]

Modified live virus (MLV) vaccines developed to protect against PRRSV circulating in North America (NA) offer limited protection to highly pathogenic (HP) PRRSV strains that are emerging in Asia. MLV vaccines specific to HP-PRRSV strains commercially available in China provide protection to HP-PRRSV; however, the efficacy of these HP-PRRSV vaccines to current circulating NA PRRS viruses has not been reported. The aim of this study is to investigate whether pigs vaccinated with attenuated Chinese HP-PRRSV vaccine (JXA1-R) are protected from infection by NA PRRSV strain NADC-20. We found that pigs vaccinated with JXA1-R were protected from challenges with HV-PRRSV or NADC-20 as shown by fewer days of clinical fever, reduced lung pathology scores, and lower PRRS virus load in the blood. PRRSV-specific antibodies, as measured by IDEXX ELISA, appeared one week after vaccination and virus neutralizing antibodies were detected four weeks post vaccination. Pigs vaccinated with JXA1-R developed broadly neutralizing antibodies with high titers to NADC-20, JXA1-R, and HV-PRRSV. In addition, we also found that IFN-α and IFN-β occurred at higher levels in the lungs of pigs vaccinated with JXA1-R. Taken together, our studies provide the first evidence that JXA1-R can confer protection in pigs against the heterologous NA PRRSV strain NADC-20. [ABSTRACT FROM AUTHOR]

In Korea, porcine reproductive and respiratory syndrome (PRRS) is caused by European (type 1) and North American (type 2) strains of PRRS virus (PRRSV). In the present study, the efficacy of a multi-strain PRRSV vaccine inactivated with binary ethylenimine (BEI) was evaluated in pigs. The vaccine contained one type 1 strain (GCEU0907) and two type 2 strains (GC4019 and GC6262). Three vaccinated groups (four pigs per group) and three mock vaccinated groups (four pigs per group) were challenged with infectious PRRSV (strains GC4019, GC6262 or GCEU0907), then euthanased at 28 days post-infection. Mean anti-PRRSV neutralising antibody titres were significantly higher in the vaccinated groups than in the mock vaccinated groups. Mean blood virus titres in the mock vaccinated groups were significantly higher than those in the vaccinated groups from 5 to 28 days post-infection. On pathological examination, there were less severe macroscopic and microscopic lesions in vaccinated pigs compared with mock vaccinated pigs. [ABSTRACT FROM AUTHOR]

Immunogenicity of protein subunit vaccines may be dramatically improved by targeting them through antibodies specific to c-type lectin receptors (CLRs) of dendritic cells in mice, cattle, and primates. This novel vaccine development approach has not yet been explored in pigs or other species largely due to the lack of key reagents. In this study, we demonstrate that porcine reproductive and respiratory syndrome virus (PRRSV) antigen was targeted efficiently to dendritic cells through antibodies specific to a porcine CLR molecule DC-SIGN (dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin) in pigs. A recombinant PRRSV antigen (shGP45M) was constructed by fusing secretory-competent subunits of GP4, GP5 and M proteins derived from genetically-shuffled strains of PRRSV. In vaccinated pigs, when the PRRSV shGP45M antigen was delivered through a recombinant mouse-porcine chimeric antibody specific to the porcine DC-SIGN (pDC-SIGN) neck domain, porcine dendritic cells rapidly internalized them in vitro and induced higher numbers of antigen-specific interferon-γ producing CD4T cells compared to the pigs receiving non-targeted PRRSV shGP45M antigen. The pDC-SIGN targeting of recombinant antigen subunits may serve as an alternative or complementary strategy to existing vaccines to improve protective immunity against PRRSV by inducing efficient T cell responses. [ABSTRACT FROM AUTHOR]

The porcine reproductive and respiratory syndrome virus (PRRSV) is a rapidly evolving and diversifying pathogen necessitating the development of improved vaccines. Immunity to PRRSV is not well understood although there are data suggesting that virus-specific T cell IFN-γ responses play an important role. We therefore aimed to better characterise the T cell response to genotype 1 (European) PRRSV by utilising a synthetic peptide library spanning the entire proteome and a small cohort of pigs rendered immune to PRRSV-1 Olot/91 by repeated experimental infection. Using an IFN-γ ELISpot assay as a read-out, we were able to identify 9 antigenic regions on 5 of the viral proteins and determine the corresponding responder T cell phenotype. The diversity of the IFN-γ response to PRRSV proteins suggests that antigenic regions are scattered throughout the proteome and no one single antigen dominates the T cell response. To address the identification of well-conserved T cell antigens, we subsequently screened groups of pigs infected with a closely related avirulent PRRSV-1 strain (Lelystad) and a divergent virulent subtype 3 strain (SU1-Bel). Whilst T cell responses from both groups were observed against many of the antigens identified in the first study, animals infected with the SU1-Bel strain showed the greatest response against peptides representing the non-structural protein 5. The proteome-wide peptide library screening method used here, as well as the antigens identified, warrant further evaluation in the context of next generation vaccine development. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant reproductive losses in the sow herd and respiratory disease in growing pigs. The virus belongs to the family Arteriviridae and there are two major genotypes. Type 1 is represented by Lelystad virus, the European prototype virus, and Type 2 is represented by the North American prototype virus, VR-2332. Depending on husbandry, immune status of the herd, and virulence of the isolate, the severity of disease and magnitude of economic loss can be variable. Vaccine use is not always successful indicating a lack of cross-protection between vaccine strains and circulating wild-type viruses. To date, there is no clear method to demonstrate if a vaccine confers protection against a specific isolate except for empirical animal studies. In 2006, a new lineage of Type 2 PRRSV emerged in Chinese swine herds that were suffering dramatic losses resulting in those viruses being described as “Highly Pathogenic PRRSV” (HP-PRRSV). Experimental reproduction of severe disease with HP-PRRSV isolates and virus derived from HP-PRRSV clones demonstrated the causal role of this virus. Recently, partial heterologous protection has been reported for Type 1 and Type 2 attenuated PRRSV vaccines against challenge by different Chinese HP-PRRSV isolates providing some hope for reducing economic loss. This paper reports the efficacy of a commercially available Type 2 attenuated vaccine in young pigs against heterologous challenge with a Chinese and Vietnamese HP-PRRSV isolate. When compared to unvaccinated pigs, vaccination decreased the length of viremia and viral titer, diminished the time of high fever and reduced macroscopic lung scores following homologous and heterologous PRRSV challenge. These results demonstrate the potential use of vaccine as an aid in the control of HP-PRRSV outbreaks. [ABSTRACT FROM AUTHOR]

Clinical outbreaks due to Actinobacillus pleuropneumoniae occur recurrently, despite the wide-scale use of antimicrobials or vaccination. Therefore, new approaches for the prevention and control of these outbreaks are necessary. For the development of alternative measures, more insight into the transmission of the bacterium on farms is necessary. The aim of this cohort study was to quantify transmission of A. pleuropneumoniae amongst weaned piglets on farms. We investigated three possible transmission routes: (i) indirect transmission by infected piglets within the same compartment, (ii) transmission by infected pigs in adjacent pens and (iii) transmission by direct contact within pens. Additionally, we evaluated the effect of independent litter characteristics on the probability of infection. Two farms participated in our study. Serum and tonsil brush samples were collected from sows pre-farrowing. Serum was analysed for antibodies against Apx toxins and Omp. Subsequently, tonsil brush samples were collected from all piglets from these dams ( N = 542) in three cohorts, 3 days before weaning and 6 weeks later. Tonsil samples were analysed by qPCR for the presence of the apxIVA gene of A. pleuropneumoniae . Before weaning, 25% of the piglets tested positive; 6 weeks later 47% tested positive. Regression and stochastic transmission models were used to assess the contribution of each of the three transmission routes and to estimate transmission rates. Transmission between piglets in adjacent pens did not differ significantly from that between non-adjacent pens. The transmission rate across pens was estimated to be 0.0058 day −1 (95% CI: 0.0030–0.010), whereas the transmission rate within pens was ten times higher 0.059 day −1 (95% CI: 0.048–0.072). Subsequently, the effects of parity and serological response of the dam and litter age at weaning on the probability of infection of pigs were evaluated by including these into the regression model. A higher dam ApxII antibody level was associated with a lower probability of infection of the pig after weaning; age at weaning was associated with a higher probability of infection of the pig after weaning. Finally, transmission rate estimates were used in a scenario study in which the litters within a compartment were mixed across pens at weaning instead of raising litter mates together in a pen. The results showed that the proportion of infected piglets increased to 69% if litters were mixed at weaning, indicating that farm management measures may affect spread of A. pleuropneumoniae . [ABSTRACT FROM AUTHOR]

Abstract: In veterinary animal species, vaccines are the primary tool for disease prevention, a key tool for treatment of infection, and essential for helping maintain animal welfare and productivity. Traditional vaccine development by trial-and-error has achieved many successes. However, effective vaccines that provide solid cross-protective immunity with excellent safety are still needed for many diseases. The path to development of vaccines against difficult pathogens requires recognition of uniquely evolved immunological interactions of individual animal hosts and their specific pathogens. Here, general principles that currently guide veterinary immunology and vaccinology research are reviewed, with an emphasis on examples from swine. Advances in genomics and proteomics now provide the community with powerful tools for elucidation of regulatory and effector mechanisms of protective immunity that provide new opportunities for successful translation of immunological discoveries into safe and effective vaccines. [Copyright &y& Elsevier]

Abstract: Outbreaks of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) have led to large economic losses and, subsequently, have drawn great attention to its diagnosis and prevention. To facilitate rapid discrimination of HP-PRRSV from classical PRRSV (C-PRRSV), we developed a one-step RT-PCR assay. Primer specificities were evaluated with RNA extracted from 8 viral strains and our results revealed that the primers had a high specificity for PRRSV. The assay sensitivity was 25copies/μL for both HP-PRRSV and C-PRRSV. A total of 929 serum samples were identified, of which 20.45% were HP-PRRSV-positive and 1.51% were C-PRRSV-positive, which was completely consistent with that of immunochromatochemistry and sequencing method. The proposed assay can detect the virus 2days prior the onset of symptoms and it can be performed in 2h, thereby providing a rapid method to discriminate HP-PRRSV from C-PRRSV for the identification and prevention of PRRSV infections. [Copyright &y& Elsevier]

The pseudorabies virus (PRV) is a major viral disease that causes huge economic loss in the pig industry globally. Most viruses have been found to generate anti-apoptotic factors that facilitate cell survival in the early stages of infection. This study aimed to investigate the anti-apoptotic effects of PRV and study the underlying mechanisms in the early stage of infection. We investigated and compared whether the two PRV Us3 isoforms, Us3a and Us3b, could block apoptosis induced by virus infection, and further identified molecules involved in the signaling pathways. Our results demonstrated that PRV elicits 3-phosphoinositide dependent protein kinase-1/phosphatidylinositide 3-kinases/Akt (PDK-1/PI3-K/Akt)- and nuclear factor-κB (NF-κB)-dependent signaling in the early stage of infection. Inhibition of the PI3-K/ Akt or NF-κB pathway enhanced cell death but no effect was observed on virus replication or PRV gene expression. Transiently-expressed GFP- or His-tagged PRV Us3a and Us3b cDNA protect cells against PRV-, avian reovirus- or bovine ephemeral fever virus-induced apoptosis in the cell lines. Us3a and Us3b transient over-expression upregulated several anti-apopototic signaling events, and the anti-apoptosis activity of Us3a is greater than that of Us3b. Kinase activity-deficient point or double point mutated Us3a lost the kinase activity of Us3a, which showed that kinase activity is required for the anti-apoptosis effect of Us3. Akt and NF-κB activation still occurred in UV-inactivated PRV- and cycloheximide-treated cells. In vivo study showed that PRV-infected trigeminal ganglion increases the expression of anti-apoptosis signaling molecules, including Akt, PDK-1 and IκBa, which is a similar result to that seen in the in vitro experiments. Our study suggests that signaling mechanisms may play important roles in PRV pathogenesis. [ABSTRACT FROM AUTHOR]

Highlights: [•] We showed efficacy of attenuated S. Typhimurium as mucosal vaccine in pigs. [•] We provided information contributing to control salmonellosis in pigs and humans. [•] We provided insights of a model of enteric salmonellosis. [Copyright &y& Elsevier]

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is difficult to control due to a high mutation rate and the emergence of virulent strains. The objective of this study was to analyze the immunological and pathological responses after infection with the European subtype 3 strain Lena in comparison to subtype 1 strains Belgium A and Lelystad-Ter Huurne (LV). Sixteen pigs were inoculated per strain, and sixteen pigs with PBS. At days 7 and 21 post-inoculation (p.i.), four pigs per group were immunized with an Aujeszky disease vaccine (ADV) to study the immune competence after PRRSV infection. Infection with the Lena strain resulted in fever and clinical signs. This was not observed in the Belgium A or LV-infected pigs. Infection with the Lena strain resulted in high virus titers in serum, low numbers of IFN-γ secreting cells, a change in leukocyte populations and a delayed antibody response to immunization with ADV. Levels of IL-1β, IFN-α, IL-10, IL-12, TNF-α and IFN-γ mRNA of the Lena-infected pigs were increased during the first week of infection. For pigs infected with the Belgium A or LV strain, the effects of infection on these parameters were less pronounced, although for the Belgium A-infected pigs, the level of the analyzed cytokines, except for TNF-α, and leukocyte populations were comparable to the Lena-infected pigs. These results suggest that while the outcome of infection for the three strains was comparable, with mostly clearance of viremia at day 33 p.i, differences in immune responses were observed, perhaps contributing to their virulence. [Copyright &y& Elsevier]

Abstract: The Influenza Pandemic (H1N1/09) virus was first reported in humans in Mexico in April 2009 and a pandemic level was declared on 11th of June 2009 by the World Health Organization (). Public misconceptions about the transmission of H1N1/09 were caused by the inadequate naming of the disease as ‘swine influenza’. This cross-sectional study was conducted at the height of the outbreak in the Australian human population and before the virus was reported in the first piggery in Australia in July 2009 (). The aims of this study were to evaluate pig producers’ perceptions about the virus and the outbreak financial impact and influence on on-farm biosecurity practices. A questionnaire was designed and posted to Australian Pork Limited (APL) members (n =460), obtaining responses from 182 producers (39.6%). Pig producers had good general knowledge on potential transmission pathways for H1N1/09 between people, with direct or close contact with a sick person perceived as the most likely pathways. Changes on biosecurity practices, such as asking visitors if they had recently been overseas (27.8%) and not allowing any visitor to inspect their pigs (18.3%), were reported among respondents. In addition, approximately 40% of producers asked their employees to notify flu like symptoms, consulted a veterinarian on H1N1/09 and visited websites to seek information on H1N1/09. A higher adoption of these practices was observed among large (>100 sows) than small herds. Only 2.9% of respondents reported a reduction in pig sales during the outbreak. However, approximately one third of producers reported being financially and emotionally stressed, 38.2% were distressed about the health of their pigs and 16.7% about their own health. The most important sources of information were APL (93%), veterinarians (89%) and the state Department of Primary Industries (DPI) (75%). The first two considered the most trusted sources of information. Television, radio and other farmers were considered more important sources of information by small herds and veterinarians by larger herds. Producers believed that the H1N1/09 outbreak was better managed by the pork industry (89.9%) than by the health authorities (58.8%), and the on-going communication with APL was the main strength of the outbreak management. Communication and extension programs in future outbreaks should consider the needs of all sectors of the pig industry to increase their effectiveness. [Copyright &y& Elsevier]

Abstract: Taenia solium causes cysticercosis in pigs and taeniasis and neurocysticercosis in humans. Oncosphere antigens have proven to be effective as vaccines to protect pigs against an experimental infection with T. solium. A pair-matched vaccination trial field, using a combination of two recombinant antigens, TSOL16 and TSOL18, was undertaken in rural villages of Peru to evaluate the efficacy of this vaccine under natural conditions. Pairs of pigs (n =137) comprising one vaccinated and one control animal, were allocated to local villagers. Animals received two vaccinations with 200μg of each of TSOL16 and TSOL18, plus 5mg Quil-A. Necropsies were performed 7 months after the animals were distributed to the farmers. Vaccination reduced 99.7% and 99.9% (p <0.01) the total number of cysts and the number of viable cysts, respectively. Immunization with the TSOL16–TSOL18 vaccines has the potential to control T. solium transmission in areas where the disease is endemic, reducing the source for tapeworm infections in humans. [Copyright &y& Elsevier]

Abstract: The current study evaluates combined aerosol-vaginal delivery of a MOMP-based Chlamydia trachomatis (serovar E) DNA vaccine in a pig genital challenge model. Most non-replicating antigens are rather poor mucosal immunogens in comparison to replicating antigens. Therefore, a mucosal administered DNA vaccine, which actually mimics a live vaccine, could be promising. Protection was promoted by plasmids encoding the porcine granulocyte macrophage-colony stimulating factor (pcDNA3.1zeo::GM-CSF), the Escherichia coli thermo-labile enterotoxin (LT) subunit A (plasmid PJV2004::LTa) and subunit B (plasmid PJV2005::LTb). Mucosal C. trachomatis DNA vaccination induced significant protection against genital C. trachomatis challenge although the infection could not be eradicated. Intradermal immunization was significantly less efficient in protecting experimentally infected pigs. Protection was correlated with efficient T cell priming and significantly higher serum IgA titers following primo vaccination. [Copyright &y& Elsevier]

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) shows highly restricted cell tropism and targets subpopulations of differentiated macrophages such as porcine alveolar macrophages (PAM) in the natural host. Although primary PAM cells would be ideal for in vitro virus production, they are not only difficult and expensive for establishment but cannot be frozen reliably for long-term storage and use. Apart from PAM cells, African green monkey kidney derived Marc-145 cells are used commonly for virus propagation. However, concerns have been raised regarding a possible modification of specific epitopes associated with virus neutralization because of distinct virus entry between PAM and Marc-145 cells. In order to overcome these problems, the present study was aimed to generate immortalized porcine monocyte/macrophage cell lines and to evaluate their potential for PRRSV production. Primary PAM cells were transfected stably with the human telomerase reverse transcriptase (hTERT) cDNA by a retrovirus vector so that constitutive expression of the hTERT protein allows cells to proliferate indefinitely. The newly immortalized PAM clones were shown to exert functional telomerase activity, indicating sustained expression of hTERT. In addition, telomerase-immortalization of PAMs did not affect expression levels of the native CD163 receptor on their surface. It was further demonstrated that these continuous PAM cell lines are fully permissive for the efficient growth of both type 1 and 2 PRRSV strains. The findings suggest that the hTERT-immortalized PAM cell lines can enable us to facilitate the continued use of PAMs for virus isolation and production and to provide a promising tool for viral pathogenesis and immune function studies. [Copyright &y& Elsevier]

Abstract: Lawsonia intracellularis is an intracellular bacterium causing proliferative enteropathy in various animal species, and is considered an economically important pathogen of pigs. Rats and mice have been implicated as external vectors for a wide range of pig pathogens, including L. intracellularis. Previous studies have demonstrated L. intracellularis infection and proliferative enteropathy in rodents, but did not show the duration of shedding or the number of L. intracellularis shed by infected rodents, and therefore the infection risk that rodents pose to pigs. In this study, the number of L. intracellularis shed in the faeces and intestinal mucosa of wild rats trapped on pig farms was determined by a quantitative real time polymerase chain reaction assay. The prevalence of L. intracellularis in wild rats trapped on pig farms with endemic proliferative enteropathy (PE) was very high (≥70.6%), and large numbers of L. intracellularis were shed (1010/g of faeces) in a small proportion of wild rats. The duration of colonisation in laboratory rats and mice challenged with porcine isolates of L. intracellularis was also shown. Faecal shedding of L. intracellularis persisted for 14–21 days in rats and mice that were mildly affected with histological lesions of PE. The humoral immune response to L. intracellularis persisted for 40 days in both species. This study demonstrates that rodents may be an important reservoir of L. intracellularis on piggeries, and hence rodent control is important in disease eradication programs on pig farms. [Copyright &y& Elsevier]

Abstract: We aim to develop a method that confers rapid protection against foot-and-mouth disease (FMD) by passive immunization with recombinant llama single-domain antibody fragments (VHHs). Previously constructed genetic fusions of two VHHs (VHH2s) that either neutralizes FMDV or binds to porcine immunoglobulin to increase the serum half-life, conferred only limited protection to pigs. We therefore now generated VHH3s containing an additional FMDV binding VHH. Two VHH3s neutralized FMDV more potently than single VHHs and were highly produced by yeast cells. Injection of a mixture of these two VHH3s 24h before FMD challenge infection of pigs reduced and delayed the development of clinical disease, viraemia and viral shedding. Furthermore, it significantly (P =0.023) delayed FMD transmission. Thus, we have shown a proof of concept of passive FMD immunoprophylaxis using VHHs. [Copyright &y& Elsevier]

Abstract: To develop a more efficacious human adenovirus (Ad5)-vectored foot-and-mouth disease virus (FMDV) subunit vaccine (Ad5-A24) we have included coding regions for FMDV nonstructural proteins 2B and 2C. These proteins are involved in membrane re-arrangements resulting in the proliferation of cytoplasmic vesicles which serve as the sites of virus replication. Cells infected with a vector containing full-length 2B (Ad5-CI-A24-2B) had a significant increase in the number of cytoplasmic vesicles as compared to cells infected with the original vector or a vector containing full-length 2BC. Swine inoculated with Ad5-CI-A24-2B developed an enhanced FMDV-specific neutralizing antibody response as compared to animals inoculated with the original vector and showed no clinical signs of disease after challenge. In a second experiment animals vaccinated with Ad5-CI-A24-2B were not fully protected but had a more rapid and robust humoral response and two out of three pigs had delayed and less severe disease than animals in the other vaccinated groups. These results suggest that incorporation of the complete coding region of 2B into the vaccine enhances its potency and protective efficacy. [Copyright &y& Elsevier]

This experiment was designed to test the effect of polymorphism in the cholesterol 7-alpha hydroxylase (CYP7) gene locus and dietary cholesterol (C) on cerebrum C in neonatal pigs fed sow's milk formulas. Thirty-six pigs (18 male and 18 female) genetically selected for high (HG) or low (LG) plasma total C were weaned at 24-36 h after birth and assigned in a 2 x 2 x 2 factorial arrangement of treatments with 2 diets (0 or 0.5% C), 2 sexes, and 2 genotypes (HG and LG). Individually housed pigs consumed diets ad libitum for 42 d. Open-field behavior was tested at wk 2 and 4. All pigs were killed at 42 d of age, the cerebrum was weighed, and C content and concentration measured. All data were analyzed by general linear model ANOVA. Cerebrum weight was greater in HG than LG pigs (P < 0.03) but was not affected by diet or sex. Pigs fed C tended to have a higher cerebrum C concentration than those deprived (P = 0.12). At 2 wk, LG pigs explored a novel open-field environment less often (P < 0.001) than did HG pigs. At 4 wk, some LG pigs explored the open field but fewer (P < 0.001) vs. HG pigs retreated back to the safe area. There were no genotype x diet, genotype x sex, or diet x sex interactions affecting cerebrum weight, or C content or concentration. Polymorphism in the CYP7 gene locus affected cerebrum weight and behavior and dietary C tended to increase cerebrum C concentration in neonatal pigs. These findings in neonatal pigs have considerable potential importance in human infant nutrition and behavioral development. [ABSTRACT FROM AUTHOR]

Abstract: In the present study, an immunochromatographic strip was developed for the serological detection of trichinellosis in swine. In the strip, the excretory–secretory (ES) antigens of Trichinella labelled with colloidal gold was used as the detector, and the staphylococcal protein A (SPA) and goat anti-ES antibody were blotted on the nitrocellulose membrane for the test and control lines, respectively. The evaluation of the strip was performed by comparing 60 clinical positive blood samples detected by the artificial digestion method with 46 serum samples from pigs infected with parasites other than Trichinella and 30 serum samples of parasite-free healthy pigs. The strip was shown to be of high specificity and sensitivity that were closely correlated with those of ELISA. Furthermore, the dipstick assay based on the strip is rapid (10min) and easy to perform with no requirement of special skill, reagent or equipment. This suggests the immunochromatographic strip is an acceptable alternative to be used in clinical laboratories lacking specialized equipment as well as for field diagnosis. [Copyright &y& Elsevier]