Porcine reproductive and respiratory syndrome virus NADC30-like strain accelerates Streptococcus suis serotype 2 infection in vivo and in vitro.
Li Jianda,Wang Jinbao,Liu Yueyue,Yang Jie,Guo Lihui,Ren Sufang,Chen Zhi,Liu Zhaoshan,Zhang Yuyu,Qiu Wenbin,Li Yubao,Zhang Shujin,Yu Jiang,Wu Jiaqiang
Transboundary and emerging diseases
Porcine reproductive and respiratory syndrome (PRRS), an economically significant pandemic disease, commonly results in increased impact of bacterial infections, including those by Streptococcus suis (S. suis). In recent years, PRRS virus (PRRSV) NADC30-like strain has emerged in different regions of China, and coinfected with S. suis and PRRSV has also gradually increased in clinical performance. However, the mechanisms involved in host innate responses towards S. suis and their implications of coinfection with NADC30-like strain remain unknown. Therefore, the pathogenicity of NADC30-like strain and S. suis serotype 2 (SS2) coinfection in vivo and in vitro was investigated in this study. The results showed that NADC30-like increased the invasion and proliferation of SS2 in blood and tissues, resulting in more severe pneumonia, myocarditis, and peritonitisas well as higher mortality rate in pigs. In vitro, NADC30-like strain increased the invasion and survival of SS2 in porcine alveolar macrophages (PAM) cells, causing more drastic expression of inflammatory cytokines and activation of NF-ĸB signalling. These results pave the way for understanding the interaction of S. suis with the swine immune system and their modulation in a viral coinfection.
Porcine sialoadhesin suppresses type I interferon production to support porcine reproductive and respiratory syndrome virus infection.
Liu Yingqi,Li Rui,Qiao Songlin,Chen Xin-Xin,Deng Ruiguang,Zhang Gaiping
Porcine reproductive and respiratory syndrome virus (PRRSV) is a significant threat to the global swine industry. Porcine sialoadhesin (poSn) has been previously shown to mediate PRRSV attachment and internalization. In the current study, we report its unidentified role in antagonism of type I interferon (IFN) production during PRRSV infection. We determined that poSn facilitated PRRSV infection via inhibition of type I IFN transcription. Mechanistically, poSn interacted with a 12 kDa DNAX-activation protein (DAP12), which was dependent on residues 51-57 within DAP12 transmembrane domain (TMD). PRRSV exploited the poSn-DAP12 pathway to attenuate activation of nuclear factor-kappa B (NF-κB). More importantly, the poSn-DAP12 pathway was involved in inhibiting poly (I:C)-triggered IFN production. All these results reveal a novel role of poSn in suppressing host antiviral responses, which deepens our understanding of PRRSV pathogenesis.
A potential endemic strain in China: NADC34-like porcine reproductive and respiratory syndrome virus.
Xu Hu,Song Shuaijie,Zhao Jing,Leng ChaoLiang,Fu Jun,Li Chao,Tang Yan-Dong,Xiang Lirun,Peng Jinmei,Wang Qian,Zhao Hongyuan,An Tongqing,Cai XueHui,Zhang HongLiang,Tian Zhi-Jun
Transboundary and emerging diseases
Porcine respiratory and reproductive syndrome virus (PRRSV) causes an economically important disease affecting commercial pork production worldwide. NADC34-like PRRSV has had a strong impact on the U.S. and Peruvian pig industries in recent years and also emerged in northeastern China in 2017. However, the endemic status of NADC34-like PRRSV in China is unclear. In this study, we examined 650 tissue samples collected from 16 Provinces in China from 2018 to 2019. Six NADC34-like PRRSV strains were detected in samples from three Provinces, and the complete genomes of four of these strains were sequenced. Phylogenetic analysis showed that these novel PRRSV strains belong to sublineage 1.5 (or NADC34-like PRRSV), forming two groups in China. Sequence alignment suggested that these novel strains share the same 100-aa deletion in the Nsp2 protein that was identified in IA/2014/NADC34 isolated from the United States in 2014. Recombination analysis revealed that five of eight complete genome sequences are derived from recombination between IA/2014/NADC34 and ISU30 or NADC30. The number and distribution of NADC34-like PRRSVs is increasing in China. Importantly, compared with the currently endemic strain NADC30-like PRRSV, NADC34-like PRRSV has the potential to be an endemic strain in China. This study will help us understand the epidemic status of NADC34-like PRRSV in China and provide data for further monitoring this type of PRRSV in China.
Thyroid hormone suppression in feeder pigs following polymicrobial or porcine reproductive and respiratory syndrome virus-2 challenge.
Pasternak J Alex,MacPhee Daniel J,Lunney Joan K,Rowland Raymond R R,Dyck Michael K,Fortin Frédéric,Dekkers Jack C M,Plastow Graham S,Harding John C S,
Journal of animal science
Thyroid hormones are powerful regulators of growth, development, and basal metabolic rate and can be dysregulated under conditions of severe stress or illness. To understand the role of these hormones in porcine disease response, serum samples were obtained from three batches of nursery-aged pigs (n = 208) exposed to a natural polymicrobial disease challenge with an array of bacterial and viral pathogens. Levels of total thyroxin (T4) and triiodothyronine (T3) assessed in sera by radioimmunoassay, decreased significantly by 14 days post-exposure (DPE). Levels of T3 partially rebounded by 48 DPE, while T4 levels remain depressed. Post-exposure T3 and T4 levels were positively correlated with acute and long-term average daily gain (ADG). Cross-sectional sampling of animals maintained at the high health source farms, showed no equivalent change in either hormone when managed under standard industrial conditions. To further elucidate the effect of porcine reproductive and respiratory syndrome virus (PRRSV)-infection on thyroid hormone levels, archived sera over 42 days post inoculation (DPI) from nursery pigs (N = 190) challenged with one of two PRRSV2 strains by the PRRS Host Genetics Consortium were similarly assessed, with animals selected in a two-by-two design, to investigate biological extremes in ADG and viral load (VL). All animals showed a similar decrease in both thyroid hormones reaching a minimum at 7 DPI and returning to near pre-challenge levels by 42 DPI. Post-challenge T3 and T4 levels were significantly greater in high ADG groups, with no significant association with VL or strain. The results of this study demonstrate porcine susceptibility to thyroid disruption in response to disease challenge and demonstrate a relationship between this response and growth performance.
In vitro and in vivo studies of deglycosylated chimeric porcine reproductive and respiratory syndrome virus as a vaccine candidate and its realistic revenue impact at commercial pig production level.
Kim Jung-Ju,Lee Jung-Ah,Choi Hwi-Yeon,Han Jang-Hyuck,Huh Won,Pi Jae-Ho,Lee Jung-Keun,Park Sangshin,Cho Ki-Hyun,Lee Joong-Bok
Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses in the swine industry worldwide. Vaccination is the most effective method to control the disease. In a previous study, a chimeric PRRSV named as K418 which had a genome composed of ORF 1 from the FL12 strain and ORF 2-7 from the Korean representative LMY strain was created. We constructed K418DM, K418 with deglycosylated glycoprotein 5 (GP5), to improve its humoral immunity. In the follow-up on in vivo and in vitro virological and serological tests, no back mutation in amino acids of GP5 associated with deglycosylation was shown after 9 passages on MARC-145 cells, whereas only one case of back mutation was detected after single passage in pig. In serological study, K418DM induced higher serum neutralization (SN) antibody and more limited viremia compared with those of K418 virus. In clinical trial and economic analysis, the K418DM elicited SN antibody titers and PRRSV-specific IgG over protection limit. From the economic viewpoint, there was statistically significant reduction in percentage of weak pigs. These results indicated that vaccination with the K418DM may provide enhanced protection for pigs in PRRS endemic situation and increase growth performance in commercial pig farms.
Development of a stochastic agent-based model to evaluate surveillance strategies for detection of emergent porcine reproductive and respiratory syndrome strains.
Arruda A G,Poljak Z,Knowles D,McLean A
BMC veterinary research
BACKGROUND:The objective of the current study was to develop a stochastic agent-based model using empirical data from Ontario (Canada) swine sites in order to evaluate different surveillance strategies for detection of emerging porcine reproductive and respiratory syndrome virus (PRRSV) strains at the regional level. Four strategies were evaluated, including (i) random sampling of fixed numbers of swine sites monthly; (ii) risk-based sampling of fixed numbers, specifically of breeding sites (high-consequence sites); (iii) risk-based sampling of fixed numbers of low biosecurity sites (high-risk); and (iv) risk-based sampling of breeding sites that are characterized as low biosecurity sites (high-risk/high-consequence). The model simulated transmission of a hypothetical emerging PRRSV strain between swine sites through three important industry networks (production system, truck and feed networks) while considering sites' underlying immunity due to past or recent exposure to heterologous PRRSV strains, as well as demographic, geographic and biosecurity-related PRRS risk factors. Outcomes of interest included surveillance system sensitivity and time to detection of the three first cases over a period of approximately three years. RESULTS:Surveillance system sensitivities were low and time to detection of three first cases was long across all examined scenarios. CONCLUSION:Traditional modes of implementing high-risk and high-consequence risk-based surveillance based on site's static characteristics do not appear to substantially improve surveillance system sensitivity. Novel strategies need to be developed and considered for rapid detection of this and other emerging swine infectious diseases. None of the four strategies compared herein appeared optimal for early detection of an emerging PPRSV strain at the regional level considering model assumptions, the underlying population of interest, and absence of other forms of surveillance.
Effect of imidacloprid ingestion on immune responses to porcine reproductive and respiratory syndrome virus.
Hernandez J,Volland A,Leyshon B J,Juda M,Ridlon J M,Johnson R W,Steelman A J
Nicotine and acetylcholine cause immunosuppresion by signaling to the α7 nicotinic acetylcholine receptor (α7 nAChR) on immune cells. Neonicotinoids are nAChR agonists and widly used insecticides. We aimed to define the immunosuppressive potential of dietary exposure to the neonicotinoid imidacloprid (IMI) on the generation of innate and adaptive immune responses to porcine reproductive and respiratory syndrome virus (PRRSV). Piglets were randomized into groups based on diet and infection. Behavioral signs of illness were recorded. Urine IMI levels were measured by high performance liquid chromatography-mass spectrometry. Flow cytometry was used to determine the expression pattern of the α7 nAChR on porcine leukocytes as well as the effects of infection and treatment on circulating leukocyte populations. Serum cytokines and PRRSV-specific antibody levels were determined by ELISA. Viral RNA in lung, spleen and plasma was determined by RT-qPCR. Pigs in the treatment group had elevated urine levels of IMI. Treatment with IMI reduced body weight, caused bouts of hypothermia, increased serum IL-10 and elevated levels of virus-specific antibodies. Viral RNA levels in the spleen showed a trend toward being increased in pigs fed IMI. Our data indicates that IMI injection may modulate virus specific immune function during PRRSV infection.
Immuno-modulating properties of Tulathromycin in porcine monocyte-derived macrophages infected with porcine reproductive and respiratory syndrome virus.
Desmonts de Lamache D,Moges R,Siddiq A,Allain T,Feener T D,Muench G P,McKenna N,Yates R M,Buret A G
Porcine reproductive and respiratory syndrome virus (PRRSV) is a positive-stranded RNA virus that grows in macrophages and causes acute pneumonia in pigs. PRRSV causes devastating losses to the porcine industry. However, due to its high antigenic variability and poorly understood immunopathogenesis, there is currently no effective vaccine or treatment to control PRRSV infection. The common occurrence of PRRSV infection with bacterial infections as well as its inflammatory-driven pathobiology raises the question of the value of antibiotics with immunomodulating properties for the treatment of the disease it causes. The macrolide antibiotic Tulathromycin (TUL) has been found to exhibit potent anti-inflammatory and immunomodulating properties in cattle and pigs. The aim of this study was to characterize the anti-viral and immunomodulating properties of TUL in PRRSV-infected porcine macrophages. Our findings indicate that blood monocyte-derived macrophages are readily infected by PRRSV and can be used as an effective cellular model to study PRRSV pathogenesis. TUL did not change intracellular or extracellular viral titers, not did it alter viral receptors (CD163 and CD169) expression on porcine macrophages. In contrast, TUL exhibited potent immunomodulating properties, which therefore occurred in the absence of any direct antiviral effects against PRRSV. TUL had an additive effect with PRRSV on the induction of macrophage apoptosis, and inhibited virus-induced necrosis. TUL significantly attenuated PRRSV-induced macrophage pro-inflammatory signaling (CXCL-8 and mitochondrial ROS production) and prevented PRRSV inhibition of non-opsonized and opsonized phagocytic function. Together, these data demonstrate that TUL inhibits PRRSV-induced inflammatory responses in porcine macrophages and protects against the phagocytic impairment caused by the virus. Research in live pigs is warranted to assess the potential clinical benefits of this antibiotic in the context of virally induced inflammation and tissue injury.
Cholesterol 25-hydroxylase is an interferon-inducible factor that protects against porcine reproductive and respiratory syndrome virus infection.
Song Zhongbao,Zhang Qiaoya,Liu Xuewei,Bai Juan,Zhao Yongxiang,Wang Xianwei,Jiang Ping
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.
Prevalence and genetic characteristics of porcine reproductive and respiratory syndrome virus in central China during 2016-2017: NADC30-like PRRSVs are predominant.
Guo Zhenhua,Chen Xin-Xin,Li Xiang,Qiao Songlin,Deng Ruiguang,Zhang Gaiping
NADC30-like strains of porcine reproductive and respiratory syndrome virus (PRRSV) were firstly reported in China in 2013. Since then, these strains have been epidemic in more than 13 provinces/regions. During 2016-2017, a total of 18 PRRSV isolates were obtained from 52 clinical samples in Henan province. Based on comparative and phylogenetic analyses of ORF5 and partial Nsp2 genes, 83.3% (15/18) isolates belonged to NADC30-like strains, and the ORF5 shared 87.4%-95.5% nucleotide identity with NADC30/JL580 and 84.2%-89.9% with JXA1/CH-1a, respectively. The genetic variation analysis showed that extensive amino acid substitutions happened in the significant regions of ORF5 including major linear antigenic epitopes (27-30aa, 37-45aa, 52-61aa) and the potential N-glycosylation sites (32-35aa). 16.7% (3/18) isolates were very close to HP-PRRSV derived attenuated strains. Moreover, these three isolates shared common residues at the positions 33D, 59 N, 164R, 196R in ORF5 and 303D, 399T, 575V, 598R, 604G in Nsp2, which were thought to be unique to modified live vaccines (MLVs) or their derivatives. Therefore, they were probably the revertants from MLVs. Our studies showed that the HP-PRRSV strains seemed to be gradually disappearing and NADC30-like strains had become the main causative agents of PRRS in central China. Comparing with HP-PRRSVs, the ORF5 of NADC30-like PRRSV strains displayed extensive amino acid mutations which may be related with immune evasion. Furthermore, the circulation of MLV derivatives in the fields made the diagnosis and control of PRRSV more complicated.
Genetic and biological characterization of a Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2) causing significant clinical disease in the field.
Kvisgaard L K,Larsen L E,Hjulsager C K,Bøtner A,Rathkjen P H,Heegaard P M H,Bisgaard N P,Nielsen J,Hansen M S
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the cause of severe reproductive and respiratory disease in swine worldwide. In Denmark, both PRRSV-1 and PRRSV-2 are circulating and approximately 35% of pig herds are seropositive for PRRSV. In November 2010, a pig herd in the Northern part of Denmark experienced an infection with PRRSV-2 with clinical signs that were much more severe than normally reported from current Danish PRRSV-2 affected herds. Due to the clinical observations of reproductive failure in sows and high mortality in piglets, it was speculated that a new, more pathogenic or vaccine evading PRRSV strain had emerged in Denmark. The overall aim of the present study was to perform a genetic and biological characterization of the virus isolated from the diseased herd. Complete genome sequencing of isolates from this herd revealed that although the case strain had some unique genetic features including a deduced 3 amino acid deletion, it was in overall very similar to the other PRRS-2 viruses circulating in Denmark. In an experimental trial in growing pigs, no overt clinical signs or pathology were observed following intranasal inoculation with the new virus isolate. Virus shedding, acute phase protein responses and serological responses were comparable to those seen after experimental challenge with a Danish PRRSV-2 reference strain isolated in 1997. Vaccination with a commercial modified live PRRSV-2 vaccine had a clear reducing effect on virus shedding, magnitude, and duration of viremia and viral load in the lungs. Overall, the results indicate that the severe disease observed in the field was contributed by additional factors in combination with the PRRS virus infection.
Genetic analysis of a porcine reproductive and respiratory syndrome virus 1 strain in China with new patterns of amino acid deletions in nsp2, GP3 and GP4.
Zhang Qiaoya,Song Zhongbao,Yu Ying,Huang Juan,Jiang Ping,Shan Hu
Porcine reproductive and respiratory syndrome virus (PRRSV) 1 and PRRSV 2 have coexisted in China for a very long time. In this study, the complete genomic characterization of a PRRSV 1 strain named KZ2018 was conducted. The results showed that it shared 88.6% identity with Lelystad virus and 81.9-90.8% identities with other Chinese PRRSV 1 strains. Further study showed that its nsp2 protein had a unique discontinuous 6-amino acid (aa) deletion (aa357-360+aa411+aa449). Additionally, its GP3 and GP4 contained a long continuous 18-aa deletion in their overlapped region, which has never been described in other Chinese PRRSV 1 isolates. Amino acid analysis of cell epitopes revealed that GP3 and GP4 were the most variable epitopes among different Chinese PRRSV 1 isolates. The results might enrich our knowledge of PRRSV 1 strains in China.
Immune system stimulation induced by porcine reproductive and respiratory syndrome virus alters plasma free amino acid flux and dietary nitrogen utilization in starter pigs1.
McGilvray Whitney D,Klein David,Wooten Hailey,Dawson John A,Hewitt Deltora,Rakhshandeh Amanda R,De Lange Cornelius F M,Rakhshandeh Anoosh
Journal of animal science
Changes in plasma free amino acid (AA) flux reflect the modification of AA metabolism in different metabolic states. Infectious diseases repartition AA away from protein retention toward processes involved in immune defense, thus impacting AA utilization in pigs. The current study sought to evaluate the effects of disease induced by a live pathogen on plasma free AA flux and whole-body nitrogen (N) utilization. Twenty gilts (BW 9.4 ± 0.9 kg) were surgically catheterized into the jugular vein, individually housed in metabolism crates, and feed-restricted (550 g/d). Intramuscular inoculation of a live field strain of porcine reproductive and respiratory syndrome virus (PRRSV) was used to induce disease. Whole-body N-balance was conducted across 3 d both before PRRSV inoculation (PRRSV-) and also after PRRSV inoculation (PRRSV+). At the end of each N-balance period, a bolus dose of a labeled [U-13C, U-15N]-AA mixture (Ile, Leu, Lys, Met, Phe, Thr, Trp, Val, and Gln) was infused intravenously, followed by serial blood collection for measurement of isotopic enrichment. A double exponential model was fitted with plasma enrichment data for each pig and each AA, and equation parameters were used to estimate plasma free AA flux and pool size. Apparent ileal digestibility (AID) of dietary N was determined using the slaughter technique and an indigestible marker. Blood chemistry, hematology, body temperature, and serum viremia indicated that PRRSV induced effective immune response in pigs (P < 0.05). Challenge with PRRSV reduced the AID of N (P < 0.05), but had no effect on apparent total tract digestibility of dietary energy (P = 0.12). Plasma flux (µmol/kg BW/h) for Met and Thr was increased by PRRSV infection (P < 0.05). A strong tendency of increased Val flux was observed in PRRSV+ pigs (P = 0.06). Infection with PRRSV increased the pool size for Lys, Met, Thr, Trp, Leu, Val, and Gln (P < 0.05). Collectively, these results suggest that PRRSV alters the utilization of dietary N and AA flux, as well as pool size, in growing pigs. The increase in Thr and Met flux in PRRSV+ pigs may be associated with enhanced utilization of these AA for the synthesis of immune system metabolites and increased catabolism of these AA. Thus, dietary Met, Thr, and Val requirements may increase in pigs infected with PRRSV, relative to the requirements for other AA.
14-3-3ε acts as a proviral factor in highly pathogenic porcine reproductive and respiratory syndrome virus infection.
Cao Shengliang,Cong Fangyuan,Tan Min,Ding Guofei,Liu Jiaqi,Li Li,Zhao Yuzhong,Liu Sidang,Xiao Yihong
The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) emerged in 2006 in China and caused great economic losses for the swine industry because of the lack of an effective vaccine. 14-3-3 proteins are generating significant interest as potential drug targets by allowing the targeting of specific pathways to elicit therapeutic effects in human diseases. In a previous study, 14-3-3s were identified to interact with non-structural protein 2 (NSP2) of PRRSV. In the present study, the specific subtype 14-3-3ε was confirmed to interact with NSP2 and play a role in the replication of the HP-PRRSV TA-12 strain. Knockdown of 14-3-3ε in Marc-145 cells and porcine alveolar macrophages (PAMs) caused a significant decrease in TA-12 replication, while stable overexpression of 14-3-3ε caused a significant increase in the replication of TA-12 and low pathogenic PRRSV (LP-PRRSV) CH-1R. The 14-3-3 inhibitor difopein also decreased TA-12 and CH-1R replication in Marc-145 cells and PAMs. These findings are consistent with 14-3-3ε acting as a proviral factor and suggest that 14-3-3ε siRNA and difopein are therapeutic candidates against PRRSV infection.
Mutations within scavenger receptor cysteine-rich (SRCR) protein domain 5 of porcine CD163 involved in infection with porcine reproductive and respiratory syndrome virus (PRRS).
The Journal of general virology
CD163, a macrophage-specific membrane scavenger receptor, serves as a cellular entry receptor for porcine reproductive and respiratory syndrome virus (PRRSV). The removal of scavenger receptor cysteine-rich (SRCR) domain 5 (SRCR5) of CD163 is sufficient to make transfected cells or genetically modified pigs resistant to PRRSV-1 and PRRSV-2 genotypes, and substitution of SRCR5 with SRCR8 from human CD163-like protein (hCD163L1) confers resistance to PRRSV-1 but not PRRSV-2 isolates. However, the specific regions within the SRCR5 polypeptide involved in PRRSV infection remain largely unknown. In this report, we performed mutational studies in order to identify which regions or amino acid sequences in the SRCR5 domain are critical for PRRSV infection. The approach used in this study was to make proline-arginine (PR) insertions along the SRCR5 polypeptide. Constructs were transfected into HEK293T cells, and then evaluated for infection with PRRSV-2 or PRRSV-1. For PRRSV-2, four PR insertions located after amino acids 8 (PR-9), 47 (PR-48), 54 (PR-55), and 99 (PR-100) had the greatest impact on infection. For PRRSV-1, insertions after amino acids 57 (PR-58) and 99 (PR-100) were critical. Computer simulations based on the crystal structure of SRCR5 showed that the mutations that affected infection localized to a similar region on the surface of the 3-D structure. Specifically, we found two surface patches that are essential for PRRSV infection. PR-58 and PR-55, which were separated by only three amino acids, had reciprocal effects on PRRSV-1 and PRRSV-2. Substitution of Glu-58 with Lys-58 reduced PRRSV-1 infection without affecting PRRSV-2, which partially explains the resistance to PRRSV-1 caused by the SRCR5 replacement with the homolog human SRCR8 previously observed. Finally, resistance to infection was observed following the disruption of any of the four conserved disulfide bonds within SRCR5. In summary, the results confirm that there are distinct differences between PRRSV-1 and PRRSV-2 on recognition of CD163; however, all mutations that affect infection locate on a similar region on the same face of SRCR5.
Porcine reproductive and respiratory syndrome virus nsp1β and nsp11 antagonize the antiviral activity of cholesterol-25-hydroxylase via lysosomal degradation.
Dong Hong,Zhou Lei,Ge Xinna,Guo Xin,Han Jun,Yang Hanchun
Porcine reproductive and respiratory syndrome virus (PRRSV) is an immunosuppressive pathogen which has been recognized to modulate the host interferon (IFN) systems. Cholesterol-25-hydroxylase (CH25 H) is an important interferon-stimulated gene (ISG)-encoded polytopic membrane protein that significantly inhibits the replication of many viruses. In the current study, we showed that PRRSV infection induced the down-regulation of the endogenous CH25H in porcine alveolar macrophages (PAMs), and then discovered that the nonstructural protein (nsp) 1β and nsp11 of PRRSV could mediate the reduction of porcine CH25H d in HEK 293FT cells. Next, the amino acids including His-159 in nsp1β, and His-129, His-144 and Lys-173 in nsp11 were determined to play crucial roles in the reduction of CH25H. Furthermore, we confirmed that the nsp1β and nsp11 mediated the degradation of CH25H by lysosomal pathway in HEK 293FT cells. Finally, it was demonstrated that the anti-PRRSV activity of CH25H could be antagonized by nsp1β and nsp11 in MARC-145 cells. Our findings suggest a manner of antagonizing the antiviral activity of CH25H by PRRSV, and provide novel insight into the understanding of PRRSV's ability of escaping the innate immunity of host.
In vitro screening antiviral activity of Thai medicinal plants against porcine reproductive and respiratory syndrome virus.
Arjin Chaiwat,Pringproa Kidsadagon,Hongsibsong Surat,Ruksiriwanich Warintorn,Seel-Audom Mintra,Mekchay Supamit,Sringarm Korawan
BMC veterinary research
BACKGROUND:Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) results in economic losses in the swine industry globally. Several studies have investigated the use of plant extracts in the prevention and control of PRRS outbreaks. Thai medicinal plants may be useful for treating PRRSV infection in pigs. Therefore, we investigated the in vitro anti-PRRSV and antioxidant properties of seven Thai medicinal plants: Caesalpinia sappan Linn., Garcinia mangostana Linn., Houttuynia cordata, Perilla frutescens, Clinacanthus nutans, Phyllanthus emblica, and Tiliacora triandra. RESULTS:Using antiviral screening, we observed that T. triandra extract strongly inhibited PRRSV infectivity in MARC-145 cells [virus titer 3.5 median tissue culture infective dose (TCID)/ml (log10)] at 24 h post-infection, whereas C. sappan extract strongly inhibited PRRSV replication [virus titer 2.5 TCID/ml (log10)] at 72 h post-infection. C. sappan extract had the highest total phenolic content [220.52 mM gallic acid equivalent/g] and lowest half-maximal inhibitory concentration [1.17 mg/ml in 2,2-diphenyl-1-picrylhydrazyl and 2.58 mg/ml in 2,2-azino-bis (3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt]. CONCLUSION:T. triandra extract could inhibit PRRSV infectivity, whereas C. sappan extract was the most effective in inhibiting PRRSV replication in MARC-145 cells. This study elucidates the antiviral activities of Thai medicinal plant extracts in vivo. The results promise that Thai medicinal plant extracts, particularly T. triandra and C. sappan extracts, can be developed into pharmaceutical drugs for the prevention of PRRS in pigs.
Nasal delivery of chitosan/alginate nanoparticle encapsulated bee (Apis mellifera) venom promotes antibody production and viral clearance during porcine reproductive and respiratory syndrome virus infection by modulating T cell related responses.
Lee Jina,Kim Yun-Mi,Kim Ju-Heon,Cho Cheong-Weon,Jeon Jong-Woon,Park Jin-Kyu,Lee Sang-Ho,Jung Bock-Gie,Lee Bong-Joo
Veterinary immunology and immunopathology
In this study, we administered specially developed chitosan/alginate nanoparticle encapsulated BV (CH/AL-BV) which has slow-releasing properties and mucosal adhesiveness to pig via nasal route and evaluate whether it can facilitate systemic immune response and improve clearance of porcine reproductive and respiratory syndrome virus (PRRSV). The CH/AL-BV-administered group with PRRSV vaccination showed significantly enhanced Th1-related responses including a high population of CD4 T lymphocyte and cytokine mRNA levels including interferon-gamma (IFN-γ) and interleukin (IL)-12 and increased PRRSV-specific IgG levels. In the PRRSV challenge experiment, the CH/AL-BV group showed a significant decrease of viral burden in the sera and tissues (lung and bronchial lymph node) and mild interstitial pneumonia signs on both lung gross examination and microscopic evaluation with high levels of PRRSV-specific IgG and viral neutralizing antibody. CH/AL-BV also effectively induced not only Th1-related immune responses including increase in portion of CD4 T lymphocyte, cytokines (IFN-γ and IL-12), and transcriptional factors (STAT4 and T-bet), but also stimulated IFN-γ-secreting cell families such as CD4 T lymphocytes and Th/memory cells. Interestingly, the CH/AL-BV group showed decrease in PRRSV-specific immune-suppressive actions, including the T regulatory cell population and its related cytokines (IL-10 and TGF-β) and transcriptional factors (STAT5 and Foxp3). Therefore, nasal-delivered CH/AL-BV may effectively induce non-specific immune stimulating actions, particularly those related to Th1 responses and viral clearance activities against PRRSV infection. Based on these results, CH/AL-BV could be a promising strategy for overcoming the disadvantages of classical PRRSV vaccination and can be applied as a preventive agent against PRRSV and other viral diseases, particularly those with immune-suppressive characteristics.
Chlorine dioxide inhibits the replication of porcine reproductive and respiratory syndrome virus by blocking viral attachment.
Zhu Zhenbang,Guo Yang,Yu Piao,Wang Xiaoying,Zhang Xiaoxiao,Dong Wenjuan,Liu Xiaohong,Guo Chunhe
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases
Porcine reproductive and respiratory syndrome virus (PRRSV) causes a great economic loss to the swine industry globally. Current prevention and treatment measures are not effective to control the outbreak and spread of porcine reproductive and respiratory syndrome (PRRS). In other words, new antiviral strategies are urgently needed. Chlorine dioxide (ClO) is regarded as a broad-spectrum disinfectant with strong inhibitory effects on microbes and parasites. The purpose of this study was to evaluate the inhibitory effects and underlying molecular mechanisms of ClO against PRRSV infection in vitro. Here, we identified ClO (the purity is 99%) could inhibit the infection and replication of PRRSV in both Marc-145 cells and porcine alveolar macrophages (PAMs). ClO could block PRRSV binding to cells rather than internalization and release, suggesting that ClO blocks the first stage of the virus life cycle. We also demonstrated that the inhibition exerted by ClO was attributed to the degradation of PRRSV genome and proteins. Moreover, we confirmed that ClO could decrease the expression of inflammatory cytokines induced by PRRSV. In summary, ClO is an efficient agent and potently suppressed PRRSV infection in vitro.
Genetic Characterization and Pathogenicity of a Novel Recombined Porcine Reproductive and Respiratory Syndrome Virus 2 among Nadc30-Like, Jxa1-Like, and Mlv-Like Strains.
Zhou Long,Kang Runmin,Yu Jifeng,Xie Bo,Chen Changying,Li Xingyu,Xie Jing,Ye Yonggang,Xiao Lu,Zhang Jinling,Yang Xin,Wang Hongning
Recombination among porcine reproductive and respiratory syndrome viruses (PRRSVs), coupled with point mutations, insertions, and deletions occurring in the genome, is considered to contribute to the emergence of new variants. Here, we report the complete genome sequences of a PRRSV field strain, designated SCN17, isolated from a RespPRRS MLV-vaccinated piglet in China in 2017. Sequence alignment revealed that SCN17 had discontinuous 131-amino acid (111 + 1 + 19-aa) deletion in the NSP2-coding region identical to that of NADC30 when compared to VR-2332. Notably, the strain, SCN17, contained an additional 1-aa deletion in NSP2, a 1-aa deletion in ORF5, and a unique 3-nt deletion in the 3'-UTR. Phylogenetic analysis showed that SCN17 clustered into NADC30-like lineage based on ORF5 genotyping, whereas it belonged to an inter-lineage between the NADC30-like and VR-2332-like lineages as established based on the full-length genome. Importantly, the SCN17 was identified as a novel virus recombined between a NADC30-like (moderately pathogenic), a JXA1-like (highly pathogenic), and an attenuated vaccine strain, RespPRRS MLV (parental strain VR-2332). Furthermore, we tested its pathogenicity in piglets. SCN17 infection caused a persistent fever, moderate interstitial pneumonia, and increased the viremia and antibody levels in the inoculated piglets. Of note, all SCN17-infected piglets survived throughout the study. The new virus was showed to be a moderately virulent isolate and have lower pathogenicity than HP-PRRSV strain, SCwhn09CD. Our results provide evidence for the continuing evolution of PRRSV field strain by genetic recombination and mutation leading to outbreaks in the vaccinated pig populations in China.
Maternally-derived neutralizing antibodies reduce vaccine efficacy against porcine reproductive and respiratory syndrome virus infection.
Renson Patricia,Fablet Christelle,Andraud Mathieu,Normand Valérie,Lebret Arnaud,Paboeuf Frédéric,Rose Nicolas,Bourry Olivier
Modified live virus (MLV) vaccines are commonly used to reduce the impact of porcine reproductive and respiratory syndrome (PRRS) but limited efficacy is achieved in field conditions. Here, we evaluated the impact of maternally-derived neutralizing antibodies (MDNAs) on vaccine efficacy after PRRS virus (PRRSV) challenge. Piglets with low (A-) or high (A+) MDNA levels derived from a commercial pig herd were moved to experimental facilities to be vaccinated (V+) or not (V-) with a PRRSV-1 MLV vaccine at 3 weeks of age (woa). Because of unexpectedly low vaccine detection in A-V+ piglets post-vaccination (pv), all V+ piglets received a second vaccination at 4 woa. Five weeks (W5) pv, piglets were inoculated with a PRRSV-1 field strain to evaluate vaccine protection, and were mingled 24 h later with non-inoculated piglets of similar immune status to assess viral transmission. Vaccine strain was detected at W2 pv in 69% and 6% of A-V+ and A+V+ piglets, and at W5 pv in 50% and 25% of A-V+ and A+V+ piglets, respectively. At W5 pv, 94% of A-V+ and 44% of A+V+ piglets seroconverted, with a significant IFNg response induction in the A-V+ group only. After challenge, compared to the V- inoculated group, viremia was 100-fold lower at 10 days post-infection in A-V+ whereas viremia was not significantly reduced in A+V+ piglets. A lower transmission rate was estimated for the A-V+ group: 0.15 [0.07-0.29] versus 0.44 [0.18-1.76] and 0.32 [0.14-0.68] for the A+V+ and V- groups, respectively. Investigations about the low vaccine strain detection after the first vaccination suggested a relationship between IFNa levels and vaccine strain detection in A-V+ piglets. We showed that MDNAs impair vaccine efficacy against PRRSV both in inoculated and contact piglets, probably by reducing vaccine replication. IFNa may also interfere with PRRSV vaccination. These new data could help improving vaccination protocols.
The effect of a porcine reproductive and respiratory syndrome outbreak on genetic parameters and reaction norms for reproductive performance in pigs1.
Putz Austin M,Schwab Clint R,Sewell Alysta D,Holtkamp Derald J,Zimmerman Jeffery J,Baker Kimberlee,Serão Nick V L,Dekkers Jack C M
Journal of animal science
The objective of this study was to estimate genetic parameters of antibody response and reproductive traits after exposure to porcine reproductive and respiratory syndrome virus. Blood samples were taken approximately 60 d after the outbreak. Antibody levels were quantified as the sample-to-positive ratio (S/P ratio) using a fluorescent microsphere assay. Reproductive traits included total number born (TNB), number born alive (NBA), number stillborn (NSB), number mummified (NBM), and number born dead (NBD). Mortality traits were log transformed for genetic analyses. Data were split into prior, during, and after the disease outbreak phases using visual appraisal of the estimates of farm-year-week effects for each reproductive trait. For NBA, data from all phases were combined into a reaction norm analysis with regression on estimates of farm-year-week effects for NBA. Heritability for S/P ratio was estimated at 0.17 ± 0.05. Heritability estimates for reproduction traits were all low and were lower during the outbreak for NBA but greater for mortality traits. TNB was not greatly affected during the outbreak, as many sows that farrowed during the outbreak were mated prior to the outbreak. Heritability for TNB decreased from 0.13 (prior) to 0.08 (after). Genetic correlation estimates between prior to and during the outbreak were high for TNB (0.86 ± 0.23) and NBA (0.98 ± 0.38) but lower for mortality traits: 0.65 ± 0.43, -0.42 ± 0.55, and 0.29 ± 1.39 for LNSB, LNBM, and LNBD, respectively. TNB prior to and after the outbreak had a lower genetic correlation (0.32 ± 0.33). In general, genetic correlation estimates of S/P ratio with reproductive performance during the outbreak were below 0.20 in absolute value, except for LNSB (-0.73 ± 0.29). Based on the reaction norm model, estimates of genetic correlations between the intercept and slope terms ranged from 0.24 ± 0.50 to 0.54 ± 0.35 depending on the parameterization used, indicating that selection for the intercept may result in indirect selection for steeper slopes, and thus, less resilient animals. In general, estimates of genetic correlations between farm-year-week effect classes based on the reaction norm model resembled estimates of genetic correlations from the multivariate analysis. Overall, compared to previous studies, antibody S/P ratios showed a lower heritability (0.17 ± 0.05) and low genetic correlations with reproductive performance during a porcine reproductive and respiratory syndrome outbreak, except for the LNSB.
Genetic diversity in envelope genes of contemporary U.S. porcine reproductive and respiratory syndrome virus strains influences viral antigenicity.
Chen Zhenhai,Collin Emily,Peddireddi Lalitha,Clement Travis,Gauger Phillip,Hause Ben M
Research in veterinary science
Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically important diseases in swine caused by porcine reproductive and respiratory syndrome virus (PRRSV). Genome sequences of sixty-six PRRSV strains were obtained using metagenomic sequencing of serum samples collected in the U.S. in 2014 to explore contemporary genetic diversity. Phylogenetic analysis of the genes encoding the envelope proteins identified four to eight distinct lineages with >87% intraclade identity. To explore the effect of the observed genetic diversity on antigenicity, the genome regions encoding either GP2a-GP3-GP4 or GP5-M in strain SD95-21 were replaced with alleles from each of eight distinct PRRSV strains using reverse genetics. The GP2a-GP3-GP4 region from only four of the eight strains yielded viable recombinant virus. When viable, both GP2a-GP3-GP4 and GP5-M variably affected antigenicity. A strain-dependent significant loss in cross reactivity was variably observed by indirect immunofluorescence assays using antisera from pigs vaccinated with commercial modified-live vaccines following replacement of GP2a-GP3-GP4 or GP5-M. Significantly reduced neutralization titers were similarly measured using antisera from naturally PRRSV-exposed pigs. These results illustrate the need to consider genomic regions besides GP5 for PRRSV epidemiology and vaccination.
Recombination in lineage 1, 3, 5 and 8 of porcine reproductive and respiratory syndrome viruses in China.
Liu Jiankui,Wei Chunhua,Lin Zhifeng,Fan Jianlin,Xia Wei,Dai Ailing,Yang Xiaoyan
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important viral swine diseases, resulting in immense economic losses in Chinese pig industry. Currently, four major lineages: lineage 1 (NADC30-like), 3 (QYYZ-like), 5.1 (VR2332-like) and 8.7 (JXA1-like) of type 2 PRRSV (North American type) have been circulating in China based on classification system, which have caused concern about the potential of virus recombination. In the present study, a novel variant of PRRSV strain named FJLIUY-2017 was isolated from abortion rate (25%) in pregnant gilts in Fujian Province in China in 2017. To further our knowledge about the novel virus strain, we characterized the complete genome of FJLIUY-2017. Comparison to PRRS sequences in GenBank confirmed the absence of close relatives (<92%), but indicated FJLIUY-2017 belonged to NADC30-like PRRSV. The full length of FJLIUY-2017 was determined to be 15017 nucleotides (nt), excluding the poly(A) tail, shared 86.2-86.6% identity with JXA1-like strains (JXA1, TJ and FJYR), 88.9-90.6% with NADC30-like PRRSVs (NADC30, FJZ03 and CHsx1401), 86.4-86.5% with VR2332-like (VR2332, RespPRRS MLV and BJ-4) and only 60.8% with LV (European type). Recombination analyses revealed genomic breakpoints in structural (ORF3, ORF4 and ORF7) and nonstructural (Nsp1, Nsp2, Nsp6, Nsp9, Nsp11 and Nsp12) regions of the genomes with evidence for recombination events between lineages 1, 3, 5.1 and 8.7. Taken altogether, the results of our study provide further confirmation that PRRSV is prone to undergo recombination events. Thus, it is critical to monitor PRRSV evolution in China and establish an effective strategy for the control of PRRS.
Assessment of area spread of porcine reproductive and respiratory syndrome (PRRS) virus in three clusters of swine farms.
Arruda A G,Sanhueza J,Corzo C,Vilalta C
Transboundary and emerging diseases
Despite decades of porcine reproductive and respiratory syndrome (PRRS) research, outbreaks with emerging and re-emerging PRRS virus (PRRSV) strains are not uncommon in North America. The role of area spread, commonly referred but not limited to airborne transmission, in originating such outbreaks is currently unknown. The main objective of this study was to explore the role of area spread on the occurrence of new PRRSV cases by combining information on genetic similarity among recovered PRRSV isolate's open-reading frame (ORF) 5 sequences and publicly available weather data. Three small regions were enrolled in the study for which high farm-level participation rate was achieved, and swine sites within those regions were readily sampled after reporting of an outbreak in a sow farm. Oral fluid PCR testing was used to determine PRRSV status of farms, and wind roses were generated for assessment of prevailing wind directions during 2-14 days preceding the outbreak. Under the conditions of this study, the data did not support the area spread theory as the main cause for these outbreaks. We suggest that for future studies, analysis of animal movement and other links between farms such as personnel, equipment and sharing of service providers should be incorporated for better insights on source of the virus. Furthermore, the development of rapid and easy diagnostic methods for ruling out resident PRRSV is urgently needed.
Molecular characterization of type 1 porcine reproductive and respiratory syndrome viruses (PRRSV) isolated in the Netherlands from 2014 to 2016.
Dortmans J C F M,Buter G J,Dijkman R,Houben M,Duinhof T F
Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of a devastating pig disease present all over the world. The remarkable genetic variation of PRRSV, makes epidemiological and molecular analysis of circulating viruses highly important to review current diagnostic tools and vaccine efficacy. Monitoring PRRS viruses supports modern herd management by explaining the source of found viruses, either internally or externally from the herd. No epidemiological or molecular study has been published on circulating PRRS-viruses in the Netherlands, since the early nineties. Therefore, the objective of this study is to investigate circulating PRRS-viruses in the Netherlands in 2014, 2015 and 2016 on a molecular level by sequencing ORF2, ORF3, ORF4, ORF5, ORF6 and ORF7. The results demonstrate that the 74 PRRSV strains belong to PRRSV-1, but the diversity among strains is high, based on nucleotide identity, individual ORF length and phylogenetic trees of individual ORFs. Furthermore, the data presented here show that the phylogenetic topology of some viruses is ORF dependent and suggests recombination. The identity of the strain of interest might be misinterpreted and wrong conclusions may be drawn in a diagnostic and epidemiological perspective, when only ORF5 is analyzed, as performed in many routine sequencing procedures.
Novel ORF5 deletion of NADC30-like porcine reproductive and respiratory syndrome viruses circulating in northern China from 2016 to 2018.
Sun Ying-Feng,Yu Hai,Jiang Xuan,Ma Ji-Fei,Xu Cheng-Qian,Yu Xiao-Xue,Li Liu-An
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
North American porcine reproductive and respiratory syndrome virus (NA-PRRSV), especially NADC30-like PRRSV, has evolved and is prevalent in China. We collected 503 samples from pig breeding farms across 4 provinces in northern China from 2016 to 2018. The samples were screened by PCR testing with specific primers that could differentiate groups of NA-PRRSV; phylogenetic trees were constructed and analyzed. Overall, 175 of 503 (34.8%) samples were positive for NA-PRRSV. Dual (NADC30-like and highly pathogenic [HP]-PRRSV; NADC30-like and typical PRRSV; HP and typical PRRSV) and triple (NADC30-like, HP, and typical PRRSV) infections (92 of 175, 52.6%) were common in coinfections by NADC30-like and HP-PRRSV. Notably, 18 of 125 (14.4%) semen samples were positive for PRRSV, and 17 of the 18 positive semen samples contained NADC30-like PRRSV. Phylogenetic analysis based on GP5 amino acids revealed that the novel NADC30-like PRRSV with a unique single amino acid deletion at position 34 has become widespread and has evolved into a new subgroup.
Damage to intestinal barrier integrity in piglets caused by porcine reproductive and respiratory syndrome virus infection.
Zhao Jin,Wan Shuangxiu,Sun Na,Sun Panpan,Sun Yaogui,Khan Ajab,Guo Jianhua,Zheng Xiaozhong,Fan Kuohai,Yin Wei,Li Hongquan
Porcine reproductive and respiratory syndrome (PRRS) induces respiratory disease and reproductive failure accompanied by gastroenteritis-like symptoms. The mechanism of intestinal barrier injury caused by PRRSV infection in piglets has yet to be investigated. An in vivo PRRSV-induced model was established in 30-day-old piglets by the intramuscular injection of 2 mL of 10 TCID/mL PRRSV for 15 days. Observations of PRRSV replication and histology were conducted in the lungs and intestine, and goblet cell counts, relative MUC2 mRNA expression, and tight junction protein, proinflammatory cytokine, TLR4, MyD88, IκB and p-IκB expression were measured. PRRSV replicated in the lungs and small intestine, as demonstrated by absolute RT-qPCR quantification, and the PRRSV N protein was detected in the lung interstitium and jejunal mucosa. PRRSV infection induced both lung and gut injury, markedly decreased villus height and the villus to crypt ratio in the small intestine, and obviously increased the number of goblet cells and the relative expression of MUC2 mRNA in the jejunum. PRRSV infection aggravated the morphological depletion of tight junction proteins and increased IL-1β, IL-6, IL-8 and TNF-α expression by activating the NF-κB signalling pathway in the jejunum. PRRSV infection impaired intestinal integrity by damaging physical and immune barriers in the intestine by inducing inflammation, which may be related to the regulation of the gut-lung axis. This study also provides a new hypothesis regarding the pathogenesis of PRRSV-induced diarrhoea.
Field evaluation of two commercial RT-rtPCR assays for porcine reproductive and respiratory syndrome virus detection using sera from ill and healthy pigs, China.
Zhang Ying-Tao,Guo Xiao-Qin,Callahan Johnny D,Yuan Gui-Li,Zhang Gui-Hong,Chen Yao,Zhang Hai-Bing,Pulscher Laura A,Lu Jia-Hai,Gray Gregory C
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Porcine reproductive and respiratory syndrome virus (PRRSV) is a highly contagious respiratory virus causing severe morbidity in pigs worldwide. Control strategies for PRRSV often rely on detecting PRRSV, culling or isolating sick pigs, disinfecting pig barns, vaccination, and monitoring for virus spread. Given the high economic impact of PRRSV on pig farms, there is a great need for rapid and reliable PRRSV detection assays. We compared the performance of 2 commercial reverse-transcription real-time PCR (RT-rtPCR) assays, the VetMAX PRRSV NA and EU reagents (ABI assay) and the PRRSV general RT-rtPCR kit (Anheal assay), for the molecular detection of PRRSV in sera collected from pigs in China. Between June and September 2015, sera were collected from 219 healthy and 104 suspected PRRSV-infected pigs on 4 farms in China. Employing blinding, the 2 assays were run by 2 laboratories (Guangzhou Animal Health Inspection Institute [GAHII] and Sun Yat-sen University [SYSU] laboratories) and compared. Although both assays detected PRRSV with 100% specificity at both laboratories, the sensitivity (95% vs. 78% at GAHII; 94% vs. 72% at SYSU Laboratory) and the reproducibility (kappa value 0.933 vs. 0.931) were slightly better for the ABI assay compared to the Anheal assay.
Genetic characterization of a novel porcine reproductive and respiratory syndrome virus type I strain from southwest China.
Zhao Jun,Zhu Ling,Deng Huidan,Li Fengqing,Xu Lei,Sun Xiangang,Yin Wenqi,Kuang Shengyao,Li Shuwei,Zhou Yuancheng,Xu Zhiwen
Archives of virology
Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating viral diseases in the global pig industry. Recently, we isolated and plaque-purified porcine reproductive and respiratory syndrome virus (PRRSV) strain SC2020-1 from "aborted piglets" on a farm in Sichuan, China. To investigate the molecular biological characteristics of this strain, it was subjected to genome sequencing and analysis. The full-length genome sequence of strain SC2020-1 was 87.7% identical to that of the Lelystad strain (PRRSV type I protoype strain) and 82.2-84.8% identical to PRRSV type I isolates from China. NSP2, ORF3, and ORF4 were the most variable regions and contained discontinuous deletions or insertions when compared to other PRRSV type I strains. Phylogenetic analysis of the complete genome sequence showed that SC2020-1 clustered with PRRSV type I but outside of the three previously described branches (Lelystad virus-like, Amervac PRRS-like, and BJEU06-1-like). The Nsp2 gene was in the same branch with EUGDHD strains from China. This is the first report of PRRSV type I infection associated with abortion in sows in southwest China. Close attention should be paid to the prevention and control of this evolving virus.
Immune responses induced by inactivated porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in neonatal pigs using different adjuvants.
Vreman Sandra,Stockhofe-Zurwieden Norbert,Popma-de Graaf Ditta J,Savelkoul Huub F J,Barnier-Quer C,Collin N,Collins Damien,McDaid Dennis,Moore Anne C,Rebel Johanna M J
Veterinary immunology and immunopathology
Vaccination of neonatal pigs could be supportive to prevent porcine reproductive and respiratory syndrome virus (PRRSV), which is an important porcine pathogen causing worldwide welfare and health problems in pigs of different age classes. However, neonatal immunity substantially differs to adults, thus different vaccines may be required in neonateal pigs. We examined if the immunogenicity and efficacy of inactivated PRRSV (iPRRSV) vaccines in neonatal pigs could be improved with adjuvants containing oil-in water (O/W) emulsions with or without Toll-like receptor (TLR) agonists and by altering the delivery route from intramuscular (i.m.) to the skin. Three-day-old PRRSV-naïve piglets (n = 54, divided in 6 groups) received a prime vaccination and a booster vaccination four weeks later. The vaccine formulations consisted of different O/W emulsions (Montanide™ ISA28RVG (ISA28)), a squalene in water emulsion (SWE) for i.m. or a Stable Emulsion (SE) with squalene for skin vaccination) and/or a mixture of TLR1/2, 7/8 and 9 agonists (TLRa) combined with iPRRSV strain 07V063. These vaccines were delivered either i.m. (ISA28, SWE, TLRa or SWE + TLRa) or into the skin (skiSE + TLRa) with dissolving microneedle (DMN)-patches. All animals received a challenge with homologous PRRSV three weeks after booster vaccination. Specific antibodies, IFN-γ production and viremia were measured at several time-points after vaccination and/or challenge, while lung pathology was studied at necropsy. After booster vaccination, only ISA28 induced a specific antibody response while a specific T-cell IFN-γ response was generated in the SWE group, that was lower for ISA28, and absent in the other groups. This suggests that prime vaccination in neonates induced a specific immune response after booster vaccination, dependent on the emulsion formulation, but not dependent on the presence of the TLRa or delivery route. Despite the measured immune responses none of the vaccines showed any efficacy. Further research focused on the early immune response in draining lymph nodes is needed to elucidate the potential of TLR agonists in vaccines for neonatal pigs.
Whole-genome sequencing and genetic characteristics of representative porcine reproductive and respiratory syndrome virus (PRRSV) isolates in Korea.
BACKGROUND:Porcine reproductive and respiratory syndrome virus (PRRSV) is a macrophage-tropic arterivirus with extremely high genetic and pathogenic heterogeneity that causes significant economic losses in the swine industry worldwide. PRRSV can be divided into two species [PRRSV1 (European) and PRRSV2 (North American)] and is usually diagnosed and genetically differentiated into several lineages based on the ORF5 gene, which constitutes only 5% of the whole genome. This study was conducted to achieve nonselective amplification and whole-genome sequencing (WGS) based on a simplified sequence-independent, single-primer amplification (SISPA) technique with next-generation sequencing (NGS), and to genetically characterize Korean PRRSV field isolates at the whole genome level. METHODS:The SISPA-NGS method coupled with a bioinformatics pipeline was utilized to retrieve full length PRRSV genomes of 19 representative Korean PRRSV strains by de novo assembly. Phylogenetic analysis, analysis of the insertion and deletion (INDEL) pattern of nonstructural protein 2 (NSP2), and recombination analysis were conducted. RESULTS:Nineteen complete PRRSV genomes were obtained with a high depth of coverage by the SISPA-NGS method. Korean PRRSV1 belonged to the Korean-specific subtype 1A and vaccine-related subtype 1C lineages, showing no evidence of recombination and divergent genetic heterogeneity with conserved NSP2 deletion patterns. Among Korean PRRSV2 isolates, modified live vaccine (MLV)-related lineage 5 viruses, lineage 1 viruses, and nation-specific Korean lineages (KOR A, B and C) could be identified. The NSP2 deletion pattern of the Korean lineages was consistent with that of the MN-184 strain (lineage 1), which indicates the common ancestor and independent evolution of Korean lineages. Multiple recombination signals were detected from Korean-lineage strains isolated in the 2010s, suggesting natural interlineage recombination between circulating KOR C and MLV strains. Interestingly, the Korean strain GGYC45 was identified as a recombinant KOR C and MLV strain harboring the KOR B ORF5 gene and might be the ancestor of currently circulating KOR B strains. Additionally, two novel lineage 1 recombinants of NADC30-like and NADC34-like viruses were detected. CONCLUSION:Genome-wide analysis of Korean PRRSV isolates retrieved by the SISPA-NGS method and de novo assembly, revealed complex evolution and recombination in the field. Therefore, continuous surveillance of PRRSV at the whole genome level should be conducted, and new vaccine strategies for more efficient control of the virus are needed.
Outbreak of Porcine Reproductive and Respiratory Syndrome Virus 1 in Taiwan.
Lin Wei-Hao,Kaewprom Kraijak,Wang Sheng-Yuan,Lin Chuen-Fu,Yang Cheng-Yao,Chiou Ming-Tang,Lin Chao-Nan
Porcine reproductive and respiratory syndrome (PRRS) causes significant economic lossesin the swine industry worldwide. The PRRS virus (PRRSV) can be divided into two species, PRRSV1 (European) and PRRSV 2 (North American). In Taiwan, PRRSV 2 isolates are dominant and causerespiratory symptoms in nursing pigs. From October to November 2018, in a pig herd in centralTaiwan, pregnant sows had abortions and stillbirths, and piglets suffered from respiratorydisorders. Laboratory tests identified the presence of PRRSV 1 in serum from sows and sucklingpiglets in this scenario. The complete genome of the identified PRRSV 1 strain was geneticallyclosely related to that of a European PRRSV vaccine strain (98.2%). This local European isolate isdesignated as PRRSV/NPUST-2789-3W-2/TW/2018 (NPUST2789). This report is the first to indicatean outbreak in Taiwan of a PRRSV 1 strain that shares a common evolutionary ancestor with theEuropean PRRSV vaccine strain.
Susceptibility of immortalized porcine kidney macrophages to porcine reproductive and respiratory syndrome virus-2 infection.
Journal of virological methods
Porcine reproductive and respiratory syndrome virus (PRRSV) displays restricted tropism to porcine alveolar macrophages in nature. Meanwhile, non-porcine cell lines derived from African green monkey kidney cell lines are permissive to PRRSV, resulting in their widespread use in PRRSV research. Furthermore, genetically modified cell lines expressing receptors targeted by PRRSV have been established. We previously established porcine immortalized kidney-derived macrophages (IPKMs) that maintained typical macrophage function. In the present study, we demonstrated the advantages of IPKMs for PRRSV research. IPKMs expressed receptors for PRRSV such as CD163 and CD169. The efficiency of virus isolation from field biological samples was higher for IPKMs than for MARC-145 cells. Five different clusters of North American type PRRSV were propagated in IPKMs. Four field strains continuously produced progeny viruses during 10 continuous passages. The efficiency of virus isolation from field biological samples and continuous progeny virus production in the sequential passages using IPKMs indicated that these cells are good vessels for PRRSV research.
High genetic diversity of Chinese porcine reproductive and respiratory syndrome viruses from 2016 to 2019.
Chen Nanhua,Xiao Yanzhao,Ye Mengxue,Li Xinshuai,Li Shubin,Xie Ningjun,Wei Yue,Wang Jialin,Zhu Jianzhong
Research in veterinary science
High genetic diversity and limited cross-protection are two major reasons for ineffective control of porcine reproductive and respiratory syndrome virus (PRRSV) infection. Therefore, it's important to dynamically monitor the prevalence of PRRSV for adopting appropriate control strategy. In this study, we analyzed PRRSV infection by detecting 712 clinical samples collected from 2016 to 2019 in China. Totally 100 samples were detected as PRRSV positive, including 2 and 98 samples were infected with PRRSV1 and PRRSV2, respectively. In addition, two out of the 98 PRRSV2 positive samples were co-infected with two distinct viruses. ORF5-based phylogenetic analysis showed that JXA1-like HP-PRRSV2 (lineage 8) and NADC30-like PRRSV2 (lineage 1) isolates are currently predominant, but QYYZ-like PRRSV2, CH-1a-like PRRSV2 and PRRSV1 isolates also co-exist in Chinese swine herds. In addition, two commercial MLV-derived viruses (TJM-F92-like and JXA1-R-like) were frequently detected. GP5 alignment also detected insertion and deletion in the extravirion domain. Our study presents the up-to-date PRRSV infection status and highlights the high genetic diversity of PRRSV currently circulating in China.
Porcine Reproductive and Respiratory Syndrome Virus strains with Higher Virulence Cause Marked Protein Profile Changes in MARC-145 Cells.
Chen Zhi,Liu Shaoning,Zhang Shujin,Zhang Yuyu,Yu Jiang,Sun Wenbo,Chen Lei,Du Yijun,Wang Jinbao,Li Yubao,Wu Jiaqiang
Porcine reproductive and respiratory syndrome is an infectious disease that causes serious economic losses to the swine industry worldwide. To better understand the pathogenesis of the porcine reproductive and respiratory syndrome virus (PRRSV), three PRRSV strains with different molecular markers and virulence were used to infect MARC-145 cells. A total of 1804 proteins were identified, and 233 altered proteins and 72 signaling pathways involved in the proteomic profiling of virus-infected MARC-145 cells increased with the virulence of the PRRSV strain. The three types of viral strains shared a common pathway-the electron transport reaction in mitochondria-in the infected-MARC-145 cells. Moreover, the antisense pathway was the most variable of all significant signaling pathways for the highly virulent SX-1 strain, indicating that this unique pathway may be connected to the high virulence of the SX-1 strain. Our study is the first attempt to provide a proteome profile of MARC-145 cells infected with PRRSV strains with different virulence, and these findings will facilitate a deep understanding of the interactions between this virus and its host.
Porcine reproductive and respiratory syndrome virus infection promotes C1QBP secretion to enhance inflammatory responses.
Li Yang,Wei Ying,Hao Wanjun,Zhao Wenkai,Zhou Yanrong,Wang Dang,Xiao Shaobo,Fang Liurong
Complement component 1, q subcomponent binding protein (C1QBP) is a receptor for the globular heads of C1q and modulates various biological processes including infection, inflammation, autoimmunity, and cancer. In our previous study to identify differentially expressed secretory proteins in Marc-145 cells infected with porcine reproductive and respiratory syndrome virus (PRRSV), mass spectrum data showed that C1QBP was secreted after PRRSV infection. However, the biological significance of secreted C1QBP remains unclear. In this study, we confirmed that PRRSV infection promoted C1QBP secretion in Marc-145 cells and porcine alveolar macrophages (PAMs), the target cells of PRRSV in vivo. Knockdown of endogenous C1QBP decreased PRRSV-induced inflammatory responses. The purified recombinant porcine C1QBP (poC1QBP) had proinflammatory effects. The exogenous addition of poC1QBP significantly enhanced PRRSV-induced inflammatory responses and abolished the inhibitory effects mediated by poC1QBP-knockdown. Taken together, these results demonstrate that PRRSV infection promotes poC1QBP secretion that enhances inflammatory responses.
Development of a biosensor from aptamers for detection of the porcine reproductive and respiratory syndrome virus.
Kuitio Chakpetch,Rasri Natchaya,Kiriwan Duangnapa,Unajak Sasimanas,Choowongkomon Kiattawee
Journal of veterinary science
BACKGROUND:Recently, the pork industry of Thailand faced an epidemic of highly virulent strains of porcine reproductive and respiratory syndrome virus (PRRSV), which spread throughout Southeast Asia, including the Lao People's Democratic Republic and Cambodia. Hence, the rapid and on-site screening of infected pigs on a farm is essential. OBJECTIVES:To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig. METHODS:New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM). RESULTS:Thirty-six candidate aptamers of the PRRSV were identified from the systematic evolution of ligands by exponential enrichment (SELEX) by capillary electrophoresis. Only 8 out of 36 aptamers could bind to the PRSSV, as shown in a colorimetric assay. Of the 8 aptamers tested, only the 1F aptamer could bind specifically to the PRSSV when presented with the classical swine fever virus and a pseudo rabies virus. The QCM was used to confirm the specificity and sensitivity of the 1F aptamer with a detection limit of 1.87 × 10 particles. CONCLUSIONS:SELEX screening of the aptamer equipped with capillary electrophoresis potentially revealed promising candidates for detecting the PRRSV. The 1F aptamer exhibited the highest specificity and selectivity against the PRRSV. These findings suggest that 1F is a promising aptamer for further developing a novel PRRSV rapid detection kit.
The T-Cell Response to Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV).
Kick Andrew R,Amaral Amanda F,Cortes Lizette M,Fogle Jonathan E,Crisci Elisa,Almond Glen W,Käser Tobias
Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause severe reproductive and respiratory pathologies resulting in immense monetary and welfare costs for the swine industry. The vaccines against PRRSV are available; but they struggle with providing protection against the plethora of heterologous PRRSV strains. To improve PRRSV vaccine development, the aim of this study was to provide an in-depth analysis of the crucial heterologous T-cell response to type-2 PRRSV. Following PRRSV modified live virus (MLV) vaccination or infection using one high- or one low-pathogenic PRRSV-strain, this nine-week study evaluated the T-cell response to different PRRSV strains. Our results demonstrate an important role for T cells in this homo- and heterologous response. Specifically, the T-helper cells were the main responders during viremia. Their peak response at 28 dpi correlated with a reduction in viremia, and their homing receptor expression indicated the additional importance for the anti-PRRSV response in the lymphatic and lung tissue. The cytocoxic T lymphocyte (CTL) response was the strongest at the site of infection-the lung and bronchoalveolar lavage. The TCR-γδ T cells were the main responders post viremia and PRRSV induced their expression of the lymph node homing the chemokine receptor, CCR7: This indicates a crucial role for TCR-γδ T cells in the anti-PRRSV response in the lymphatic system.
Effect of vaccination with a porcine reproductive and respiratory syndrome subunit vaccine on sow reproductive performance in endemic farms.
Jeong Jiwoon,Kim Seeun,Park Changhoon,Kang Ikjae,Park Kee Hwan,Ham Hee Jin,Chae Chanhee
The Veterinary record
The objective of this field study was to evaluate the reproductive performance of sows after vaccination with a porcine reproductive and respiratory syndrome (PRRS) subunit vaccine (PRRSFREE PRRS subunit vaccine, Reber Genetics, Taiwan, Republic of China) under field conditions. The study was performed in three farms with endemic infections with both PRRS virus (PRRSV)-1 and PRRSV-2, a situation representative of most Korean farms. Pregnant sows were immunised intramuscularly with 2.0 ml of the PRRS subunit vaccine at 58 and 79 days of gestation (eight and five weeks antepartum) according to the manufacturer's recommendation. Vaccination did not result in any observed adverse reaction. Vaccinated sows exhibited a significant improvement in reproductive performance (reduction of abortions) and litter characteristics (increase of weaned pigs) compared with unvaccinated sows. Vaccinated sows had significantly (P<0.05) higher PRRSV ELISA sample/positive ratio and number of PRRSV-specific interferon-γ-secreting cells compared with the unvaccinated control group. The results of this study demonstrate that the PRRS subunit vaccine can improve the reproductive performance of sows in farms with endemic PRRSV infection.
Differential evolution of antigenic regions of porcine reproductive and respiratory syndrome virus 1 before and after vaccine introduction.
Kwon Taeyong,Yoo Sung J,Lee Dong-Uk,Sunwoo Sun Young,Je Sang H,Park Jun Woo,Kim Myung-Hyee,Park Choi-Kyu,Lyoo Young S
Porcine reproductive and respiratory syndrome virus (PRRSV) is a widespread viral pathogen that has caused tremendous economic losses throughout most pig-producing countries. Nowadays, both PRRSV-1 and PRRSV-2 co-circulate in Korean pig populations, and commercial modified live vaccine (MLV) is predominantly used to control PRRS. Specifically, control strategy using only PRRSV-2 MLV that was used since 1995 cannot prevent the spread of PRRSV-1 and damage from its infection, which led to the first introduction of two additional PRRSV-1 vaccines in 2014. Despite the wide implementation with PRRSV-1 vaccines, there is a lack of knowledge about the currently circulating Korean PRRSV-1 strains. Whole structural genes of PRRSV-1 before (11) and after (17) the introduction of vaccine were compared to determine the genetic evolutionary features of PRRSV. Genetic and phylogenetic analysis indicated that Korean PRRSV-1 shared 91.5 ± 1.7% nucleotide identity but formed a unique clade based on ORF2-7 phylogeny. Bioinformatics showed increased genetic heterogeneity, enhanced diversifying selection, and the emergence of novel glycosylation sites within neutralizing epitopes of minor structural proteins after vaccine introduction. Taken together, our data provide novel insight into the evolution of minor structural proteins of PRRSV-1 in the vaccination era.
Porcine endogenous retrovirus envelope coated baculoviral DNA vaccine against porcine reproductive and respiratory syndrome virus.
Cho Yeondong,Heo Yoonki,Choi Hanul,Park Ki Hoon,Kim Sehyun,Jang Yuyeon,Lee Hee-Jung,Kim Minji,Kim Young Bong
PERV is a major virus concerning xenotransplantation study. However, the interesting part is that PERV is present in all kinds of pigs without pathogenicity and immune response. Furthermore, since pig cells have receptors for PERV, the gene delivery system using PERV envelope is highly likely to develop into an excellent viral vector in pigs. We developed a recombinant baculovirus with a modified surface for expressing the porcine endogenous retrovirus (PERV) envelope. Porcine reproductive and respiratory syndrome virus (PRRSV) infection is a severe concern in the porcine industry due to reproduction failure and respiratory symptoms. GP5 and M proteins are major immunogenic proteins of PRRSV. Using PERV-modified baculovirus (Ac mPERV) as a delivery vector, we constructed a dual antigen (GP5 and M)-encoding DNA vaccine system, Ac mPERV-C5/C6. Intramuscular immunization in mice and pigs, Ac mPERV-C5/C6 induced comparative high humoral and cellular immune responses. Our results support further development of Ac mPERV-C5/C6 as a potential PRRSV vaccine in the porcine industry. In addition, the Ac mPERV system may be applied to the generation of other effective DNA vaccines against porcine viral diseases.
Emergence of a novel highly pathogenic recombinant virus from three lineages of porcine reproductive and respiratory syndrome virus 2 in China 2017.
Chen Nanhua,Ye Mengxue,Li Shuai,Huang Yucheng,Zhou Rongyun,Yu Xiuling,Tian Kegong,Zhu Jianzhong
Transboundary and emerging diseases
A novel porcine reproductive and respiratory syndrome virus 2 (PRRSV2) was isolated from diseased piglets in Shandong, China in 2017 and denominated as SD17-38. ORF5 sequencing showed that SD17-38 contains a unique serine/asparagine deletion at position 33 and an asparagine insertion at position 60 of GP5, which has never been described. The SD17-38 complete genome was then determined, and genome-based phylogenetic analysis showed that SD17-38 is clustered with NADC30-like isolates. Sequence alignment and recombination analyses by RDP4 and SimPlot all indicated that SD17-38 is a recombinant virus from NADC30 (lineage 1), BJ-4 (lineage 5) and TJ (lineage 8) isolates. Animal challenge study in 4-week piglets showed that SD17-38 causes high fever (≥41°C), 100% morbidity and 40% mortality. In addition, significantly lower weight gain and severe histopathological lung lesions could be observed in SD17-38-infected pigs. In particular, the unique deletion and insertion in GP5 were stable during the challenge study. This study provides direct evidence for the natural occurrence of recombination events among three lineages of PRRSV2 in Chinese swine herds, resulting in the emergence of novel PRRSV variant with unique genetic property and high pathogenicity.
Optimal vaccination strategy against Mycoplasma hyopneumoniae, porcine reproductive and respiratory syndrome virus, and porcine circovirus type 2 in case of early M. hyopneumoniae infection.
Yang Siyeon,Oh Taehwan,Mago Josuke,Iwakuma Akihiro,Chae Chanhee
Veterinary medicine and science
BACKGROUND:The aim of this study was to determine the optimal vaccination strategies for the control of porcine respiratory disease complex (PRDC) caused by Mycoplasma hyopneumoniae, porcine reproductive and respiratory syndrome virus (PRRSV), and porcine circovirus type 2 (PCV2) in case of early mycoplasmal infection. METHODS:A total of 120 pigs were randomly divided into 6 groups (20 pigs per group). Four separate vaccine regimen groups were selected. Pigs from the four vaccinated groups were challenged with M. hyopneumoniae at 28 days old followed by a challenge of PRRSV or PCV2 at 49 days old. RESULTS:Regardless of PRRSV or PCV2 vaccination, pigs vaccinated with one of the M. hyopneumoniae vaccines at 7 days old had a significantly better growth performance over the whole length of the study compared to pigs vaccinated with a second M. hyopneumoniae vaccine at 21 days old. Vaccination of pigs with M. hyopneumoniae at 7 days and PRRSV at either 7, 14 or 21 days old resulted in significantly reduced PRRSV viremia and lung lesions compared to vaccination of pigs with M. hyopneumoniae and PRRSV at 21 days old. CONCLUSIONS:The efficacy of the PRRSV MLV vaccine is influenced by the different timing of M. hyopneumoniae vaccination whereas the efficacy of the PCV2 vaccine is not. This experiment study demonstrated that early vaccination with a M. hyopneumoniae vaccine should be the highest priority in order to control M. hyopneumoniae and PRRSV infection in cases of early M. hyopneumoniae infection.
Vaccination with a porcine reproductive and respiratory syndrome virus vaccine at 1-day-old improved growth performance of piglets under field conditions.
Jeong Jiwoon,Kim Seeun,Park Kee Hwan,Kang Ikjae,Park Su-Jin,Yang Siyeon,Oh Taehwan,Chae Chanhee
A porcine reproductive and respiratory syndrome virus (PRRSV) modified live-virus (MLV) vaccine was evaluated under field conditions for registration as recommended by the Republic of Korea's Animal, Plant & Fisheries Quarantine & Inspection Agency. A single dose of the vaccine was administered to 1-day-old piglets and their growth performance was monitored under field conditions. Three separate farms were selected based on their history of PRRSV-associated respiratory diseases. On each farm, 40 pigs were randomly allocated to one of two treatment groups: (i) vaccinated (n = 20) and (ii) unvaccinated (n = 20) pigs at 1 day of age. Vaccinated pigs showed an increase of their market weight of 6.23 kg/pig compared to the unvaccinated pigs (98.01 kg in vaccinated group vs. 91.78 kg in unvaccinated group; P < 0.05) and exhibited a decrease in mortality rate by 6.7% (3.3% in vaccinated group vs. 10% in unvaccinated group; P < 0.05). The pigs had a sufficiently mature immune system for the vaccine to elicit humoral and cell-mediated immunity (as measured by anti-PRRSV antibodies and PRRSV-specific interferon-γ secreting cells, respectively) at 1 day of age even in the presence of maternally derived antibodies. The results presented in this study demonstrate that the PRRSV MLV vaccine is effective in improving growth performance from day 1 all the way to day 182 in endemic farms suffering with PRRSV-2 infection or both PRRSV-1 and PRRSV-2 infection.
Investigating the relationship of porcine reproductive and respiratory syndrome virus RNA detection between adult/sow farm and wean-to-market age categories.
Jiang Yiqun,Li Qing,Trevisan Giovani,Linhares Daniel C L,MacKenzie Cameron
Porcine reproductive and respiratory syndrome (PRRS) is a disease caused by the PRRS virus (PRRSV) that has spread globally in the last 30 years and causes huge economic losses every year. This research aims to 1) investigate the relationship between the PRRSV detection in two age categories (wean-to-market and adult/sow farm), and 2) examine the extent to which the wean-to-market PRRSV positive rate forecasts the adult/sow farm PRRSV positive rate. The data we used are the PRRSV RNA detection results between 2007 and 2019 integrated by the US Swine Disease Reporting System project that represent 95% of all porcine submissions tested in the US National Animal Health Network. We first use statistical tools to investigate to what extent the increase in PRRSV positive submissions in the wean-to-market is related to the PRRSV increase in adult/sow farms. The statistical analysis confirms that an increase in the PRRSV positive rate of wean-to-market precedes the increase in the adult/sow farms to a large extent. Then we create the dynamic exponentially weighted moving average control charts to identify out-of-control points (i.e., signals) in the PRRSV rates for both wean-to-market and adult/sow farms. This control-chart-based analysis finds that 78% of PRRSV signals in the wean-to-market are followed by a PRRSV rate signal in the adult/sow farms within eight weeks. We expect that our findings will help the producers and veterinarians to justify and reinforce the implementation of bio-security and bio-contaminant practices to curb disease spread across farms.
Recombination between Vaccine and Field Strains of Porcine Reproductive and Respiratory Syndrome Virus.
Wang Anping,Chen Qi,Wang Leyi,Madson Darin,Harmon Karen,Gauger Phillip,Zhang Jianqiang,Li Ganwu
Emerging infectious diseases
We isolated and plaque purified IA76950-WT and IA70388-R, 2 porcine reproductive and respiratory syndrome viruses from pigs in the same herd in Iowa, USA, that exhibited coughing and had interstitial pneumonia. Phylogenetic and molecular evolutionary analysis indicated that IA70388-R is a natural recombinant from Fostera PRRSV vaccine and field strain IA76950-WT.
Cell-mediated immune response and protective efficacy of porcine reproductive and respiratory syndrome virus modified-live vaccines against co-challenge with PRRSV-1 and PRRSV-2.
Madapong Adthakorn,Saeng-Chuto Kepalee,Boonsoongnern Alongkot,Tantituvanont Angkana,Nilubol Dachrit
Cell-mediated immunity (CMI), IL-10, and the protective efficacy of modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccines (MLV) against co-challenge with PRRSV-1 and PRRSV-2 (HP-PRRSV) were investigated. Seventy, PRRSV-free, 3-week old, pigs were allocated into 7 groups. Six groups were intramuscularly vaccinated with MLV, including Porcilis (PRRSV-1 MLV, MSD Animal Health, The Netherlands), Amervac (PRRSV-1 MLV, Laboratorios Hipra, Spain), Fostera (PRRSV-2 MLV, Zoetis, USA), Ingelvac PRRS MLV and Ingelvac PRRS ATP (PRRSV-2, Boehringer Ingelheim, USA), and Prime Pac PRRS (PRRSV-2 MLV, MSD Animal Health, The Netherlands). Unvaccinated pigs were left as control. Lymphocyte proliferative response, IL-10 and IFN-γ production were determined. At 35 days post-vaccination (DPV), all pigs were inoculated intranasally with 2 ml of each PRRSV-1 (10 TCID/ml) and PRRSV-2 (10 TCID/ml, HP-PRRSV). Following challenge, sera were quantitatively assayed for PRRSV RNA. Pigs were necropsied at 7 days post-challenge. Viremia, macro- and microscopic lung lesion together with PRRSV antigen presence were evaluated in lung tissues. The results demonstrated that, regardless of vaccine genotype, CMI induced by all MLVs was relatively slow. Increased production of IL-10 in all vaccinated groups was observed at 7 and 14 DPV. Pigs in Amervac, Ingelvac MLV and Ingelvac ATP groups had significantly higher levels of IL-10 compared to Porcilis, Fostera and Prime Pac groups at 7 and 14 DPV. Following challenge, regardless to vaccine genotype, vaccinated pigs had significantly lower lung lesion scores and PRRSV antigens than those in the control group. Both PRRSV-1 and PRRSV-2 RNA were significantly reduced. Prime Pac pigs had lowest PRRSV-1 and PRRSV-2 RNA in serum, and micro- and macroscopic lung lesion scores (p < 0.05) compared to other vaccinated groups. In conclusion, PRRSV MLVs, regardless of vaccine genotype, can reduce viremia and lung lesions following co-challenge with PRRSV-1 and PRRSV-2 (HP-PRRSV). The main difference between PRRSV MLV is the production of IL-10 following vaccination.
Displaying epitope B and epitope 7 of porcine reproductive and respiratory syndrome virus on virus like particles of porcine circovirus type 2 provides partial protection to pigs.
Li Guopan,Liu Lei,Xu Baojuan,Hu Jixiong,Kuang Hongyan,Wang Xi,Wang Liping,Cui Xiaoxia,Sun Houmin,Rong Jun
The Journal of veterinary medical science
The Cap of porcine circovirus type 2 (PCV2) can be assembled into virus like particles (VLPs) in vitro that have multiple loops located on the particle surface. This would make it a good vehicle for displaying exogenous proteins or epitopes. We derived two epitopes, epitope B (EpB, SHIQLIYNL) and epitope 7 (Ep7, QWGRL) from Gp5 of the highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). We replaced the core region of Loop CD (LPPGGGSN) and the carboxyl terminus (KDPPL) of PCV2 Cap, respectively, to construct a bi-epitope chimeric PCV2 Cap. Its immunogenicity and protective effects were evaluated as one PRRSV subunit vaccine. The chimeric PCV2 Cap was soluble, efficiently expressed in an Escherichia coli expression system, and could be self-assembled into chimeric virus like particles (cVLPs) with a diameter of 12-15 nm. Western blotting confirmed that the cVLPs could be specifically recognized by anti-PCV2, anti-EpB and anti-Ep7 antibodies. The cVLPs vaccine could alleviate the clinical symptoms and reduce the viral loads after HP-PRRSV challenge in 100-120 days old pigs. These data suggest that the cVLPs vaccine could provide pigs with partial protection against homologous PRRSV strains, and it provides a new design for additional PRRSV subunit vaccines.
Efficacy of two porcine reproductive and respiratory syndrome (PRRS) modified-live virus (MLV) vaccines against heterologous NADC30-like PRRS virus challenge.
Chai Weidong,Liu Zhicheng,Sun Zhi,Su Liangke,Zhang Chunhong,Huang Lv
The emergence of novel and variant porcine reproductive and respiratory syndrome virus (PRRSV) strains has made controlling this disease a challenge in China. Several NADC30-like PRRSV outbreaks have occurred in mainland China since 2013. The objective of the present study was to evaluate the cross-protection efficacy of two commercial PRRS modified-live virus (MLV) vaccines, derived from classical PRRSV (VR2332) and highly pathogenic (HP) PRRSV (TJM-F92), against an increasingly circulating NADC30-like lineage in pigs. Thirty-five PRRSV- and antibody-free pigs were randomly divided into the following four groups: strict control (SC), negative control (NC), Boehringer control (BC), and Zoetis control (ZC) groups. The NADC30-like PRRSV used in this study caused fever, clinical respiratory signs, and gross and microscopic lung lesions in inoculated pigs in the NC group. Vaccination with the VR2332 vaccine significantly reduced the percentage of viremic pigs as well as gross lung lesions and improved average daily weight gain compared to the ZC and NC groups, suggesting that this MLV vaccine provides cross-protection against the NADC30-like virus. There were no significant differences in the efficacy of the two MLV vaccines based on clinical scores, immunological responses, or pathological outcomes. This study demonstrated that VR2332 MLV was effective against circulating NADC30-like PRRSV and could be used to control NADC30-like virus infections in the field.
Phylogenetics, Genomic Recombination, and NSP2 Polymorphic Patterns of Porcine Reproductive and Respiratory Syndrome Virus in China and the United States in 2014-2018.
Yu Fang,Yan Yi,Shi Mang,Liu Hai-Zhou,Zhang Hong-Liang,Yang Yong-Bo,Huang Xin-Yi,Gauger Phillip C,Zhang Jianqiang,Zhang Yan-He,Tong Guang-Zhi,Tian Zhi-Jun,Chen Jian-Jun,Cai Xue-Hui,Liu Di,Li Ganwu,An Tong-Qing
Journal of virology
Porcine reproductive and respiratory syndrome virus (PRRSV), an important pathogen that affects the pig industry, is a highly genetically diverse RNA virus. However, the phylogenetic and genomic recombination properties of this virus have not been completely elucidated. In this study, comparative analyses of all available genomic sequences of North American (NA)-type PRRSVs ( = 355, including 138 PRRSV genomes sequenced in this study) in China and the United States during 2014-2018 revealed a high frequency of interlineage recombination hot spots in nonstructural protein 9 (NSP9) and the GP2 to GP3 regions. Lineage 1 (L1) PRRSV was found to be susceptible to recombination among PRRSVs both in China and the United States. The recombinant major parent between the 1991-2013 data and the 2014-2018 data showed a trend from complex to simple. The major recombination pattern changed from an L8 to L1 backbone during 2014-2018 for Chinese PRRSVs, whereas L1 was always the major backbone for US PRRSVs. Intralineage recombination hot spots were not as concentrated as interlineage recombination hot spots. In the two main clades with differential diversity in L1, NADC30-like PRRSVs are undergoing a decrease in population genetic diversity, NADC34-like PRRSVs have been relatively stable in population genetic diversity for years. Systematic analyses of insertion and deletion (indel) polymorphisms of NSP2 divided PRRSVs into 25 patterns, which could generate novel references for the classification of PRRSVs. The results of this study contribute to a deeper understanding of the recombination of PRRSVs and indicate the need for coordinated epidemiological investigations among countries. Porcine reproductive and respiratory syndrome (PRRS) is one of the most significant swine diseases. However, the phylogenetic and genomic recombination properties of the PRRS virus (PRRSV) have not been completely elucidated. In this study, we systematically compared differences in the lineage distribution, recombination, NSP2 polymorphisms, and evolutionary dynamics between North American (NA)-type PRRSVs in China and in the United States. Strikingly, we found high frequency of interlineage recombination hot spots in nonstructural protein 9 (NSP9) and in the GP2 to GP3 region. Also, intralineage recombination hot spots were scattered across the genome between Chinese and US strains. Furthermore, we proposed novel methods based on NSP2 indel patterns for the classification of PRRSVs. Evolutionary dynamics analysis revealed that NADC30-like PRRSVs are undergoing a decrease in population genetic diversity, suggesting that a dominant population may occur and cause an outbreak. Our findings offer important insights into the recombination of PRRSVs and suggest the need for coordinated international epidemiological investigations.
First-time detection of porcine reproductive and respiratory syndrome virus (PRRSV) infection in Uruguay.
Ramos N,Mirazo S,Castro G,Cabrera K,Osorio F,Arbiza J
Transboundary and emerging diseases
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.
Using commercial ELISAs to assess humoral response in sows repeatedly vaccinated with modified live porcine reproductive and respiratory syndrome virus.
Díaz Ivan,Genís-Jorquera Blanca,Martín-Valls Gerard E,Mateu Enric
The Veterinary record
BACKGROUND:Sows in breeding herds are often mass vaccinated against porcine reproductive and respiratory syndrome (PRRS) every few months using modified live vaccines (MLV). Field veterinarians repeatedly report that multiple vaccinated sows test negative in ELISA. Obviously, this creates uncertainty when assessing the compliance of vaccination and the status of sows. METHODS:In the present study, four commercial ELISAs were used to assess the serological PRRS status in gilts and sows of three farms that were PRRS MLV vaccinated every four months. Animals were tested before vaccination (BV) and postvaccination (PV). Total and neutralising antibodies and cell-mediated responses were also measured in animals that yielded negative results in all ELISAs. RESULTS:The proportion of seronegative animals BV varied depending on the farm and the ELISA used. When samples were analysed using only one ELISA, a substantial number of negative results obtained BV remained as negative afterwards. Five animals were negative BV and PV with all the examined ELISAs. Those animals also yielded negative results in all the other immunological assays. CONCLUSION:Our findings suggest that the use of ELISA for monitoring multiple PRRS MLV vaccinated sows is very limited due to the variability of the humoral responses and the moderate agreement between tests.
Emergence of a virulent porcine reproductive and respiratory syndrome virus in Taiwan in 2018.
Lin Wei-Hao,Shih Hsing-Chun,Wang Sheng-Yuan,Lin Chuen-Fu,Yang Cheng-Yao,Chiou Ming-Tang,Lin Chao-Nan
Transboundary and emerging diseases
In March 2018, an abortion storm caused by porcine reproductive and respiratory syndrome virus was confirmed in a farrow-to-finish pig herd in Taiwan. Open reading frame 5 and non-structural protein 2 of the virus confirmed that the virus is closely related to the virulent strains circulating in the United States.
Determination of antibody induction by highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) vaccine: A comparison of two ELISA kits.
Ge Meng,Li Run-Cheng,Gong Wenjie,Tu Changchun
The Journal of veterinary medical science
Two commercial porcine reproductive and respiratory syndrome virus (PRRSV) antibody ELISA kits (IDEXX and LSI) are currently in extensive use. To determine which kit is more suitable for the evaluation of HP-PRRSV vaccine efficacy, the two kits were used to test 546 serum samples. The agreement between the results was unsatisfactory, with a kappa statistic of 0.681 and a linear correlation coefficient of 0.665. In tests of samples from experimentally vaccinated and PRRSV-negative herds, IDEXX-ELISA identified antibody-positive conversion earlier and showed a higher specificity compared to LSI-ELISA. The serological profile obtained by neutralization testing was closer to that obtained by IDEXX-ELISA than by LSI-ELISA in the late immunization period. The findings reveal that IDEXX-ELISA is the more suitable for the evaluation of antibody response to HP-PRRSV vaccine and for guiding vaccination strategies.
Aerosol Detection and Transmission of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV): What Is the Evidence, and What Are the Knowledge Gaps?
Arruda Andréia Gonçalves,Tousignant Steve,Sanhueza Juan,Vilalta Carles,Poljak Zvonimir,Torremorell Montserrat,Alonso Carmen,Corzo Cesar A
In human and veterinary medicine, there have been multiple reports of pathogens being airborne under experimental and field conditions, highlighting the importance of this transmission route. These studies shed light on different aspects related to airborne transmission such as the capability of pathogens becoming airborne, the ability of pathogens to remain infectious while airborne, the role played by environmental conditions in pathogen dissemination, and pathogen strain as an interfering factor in airborne transmission. Data showing that airborne pathogens originating from an infectious individual or population can infect susceptible hosts are scarce, especially under field conditions. Furthermore, even though disease outbreak investigations have generated important information identifying potential ports of entry of pathogens into populations, these investigations do not necessarily yield clear answers on mechanisms by which pathogens have been introduced into populations. In swine, the aerosol transmission route gained popularity during the late 1990's as suspicions of airborne transmission of porcine reproductive and respiratory syndrome virus (PRRSV) were growing. Several studies were conducted within the last 15 years contributing to the understanding of this transmission route; however, questions still remain. This paper reviews the current knowledge and identifies knowledge gaps related to PRRSV airborne transmission.
Experimental reproduction of porcine respiratory disease complex in pigs inoculated porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae and followed by inoculation with porcine circovirus type 2.
Lee Joo Young,Park Kee Hwan,Oh Taehwan,Yang Siyeon,Suh Jeongmin,Ham Hee Jin,Chae Chanhee
The Journal of veterinary medical science
The aim of this study was to reproduce severe pneumonic lesions, similar to those during naturally-occurring porcine respiratory disease complex, in pigs dually inoculated with porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae at 6 weeks of age, followed by inoculation with porcine circovirus type 2 at two weeks after. Time and sequence of infection with three pathogens mirror Asian field conditions. Microscopically, interstitial pneumonia and peribronchiolar lymphoid hyperplasia are considered the most characteristic lung lesions in infected pigs. The results of the present study demonstrate that inoculation of pigs with these three pathogens can lead to severe interstitial pneumonia with peribronchial or peribronchiolar lymphoid hyperplasia and fibrosis.
Temporal lineage dynamics of the ORF5 gene of porcine reproductive and respiratory syndrome virus in Korea in 2014-2019.
Kim Seung-Chai,Jeong Chang-Gi,Park Gyeong-Seo,Park Ji-Young,Jeoung Hye-Young,Shin Go-Eun,Ko Mi-Kyeong,Kim Seoung-Hee,Lee Kyoung-Ki,Kim Won-Il
Archives of virology
Porcine reproductive and respiratory syndrome virus (PRRSV) is the most important pathogen in the Korean swine industry. Despite efforts including improved biosecurity and vaccination protocols, the virus continues to circulate and evolve. Based on phylogenetic analysis of open reading frame 5 (ORF5), Korean PRRSVs are known to form not only globally circulating lineages but also country-specific lineages (Lin Kor A, B, and C). To understand the recent epidemiological status of PRRSV in Korea, a total of 1349 ORF5 sequences of Korean PRRSV isolates from 2014 to 2019 were analyzed. Phylogenetic analysis was conducted using the maximum-likelihood method, and temporal changes in the relative prevalence of lineages were investigated. The analysis showed that PRRSV1 and PRRSV2 were both highly prevalent throughout the years examined. Among the PRRSV1 isolates, subgroup A (90.1%) and vaccine-like subgroup C (9.0%) composed most of the population. For PRRSV2 isolates, vaccine-like lineage 5 (36.3%) was dominant, followed by Lin Kor B (25.9%), Kor C (16.6%), lineage 1 (11.6%), and Kor A (9.1%). The PRRSV2 lineage 1 population increased from 2014 (1.8%) to 2019 (29.6%) in Korea due to the continual spread of sublineage 1.8 (NADC30-like) and introduction of sublineage 1.6 into the country. Additional genetic analysis, including analysis of non synonymous and synonymous mutations, revealed evidence of diversification and positive selection in immunologically important regions of the genome, suggesting that current vaccination is failing and promoting immune-mediated selection. Overall, these findings provide insights into the epidemiological and evolutionary dynamics of cocirculating viral lineages, and constant surveillance of PRRSV occurrence is needed.
Porcine reproductive and respiratory syndrome virus dissemination across pig production systems in the United States.
Jara Manuel,Rasmussen David A,Corzo Cesar A,Machado Gustavo
Transboundary and emerging diseases
Porcine reproductive and respiratory syndrome virus (PRRSV) remains widespread in the North American pig population. Despite improvements in virus characterization, it is unclear whether PRRSV infections are a product of viral circulation within production systems (local) or across production systems (external). Here, we examined the local and external dissemination dynamics of PRRSV and the processes facilitating its spread in three production systems. Overall, PRRSV genetic diversity has declined since 2018, while phylodynamic results support frequent external transmission. We found that PRRSV dissemination predominantly occurred mostly through transmission between farms of different production companies for several months, especially from November until May, a timeframe already established as PRRSV season. Although local PRRSV dissemination occurred mainly through regular pig flow (from sow to nursery and then to finisher farms), an important flux of PRRSV dissemination also occurred in the opposite direction, from finisher to sow and nursery farms, highlighting the importance of downstream farms as sources of the virus. Our results also showed that farms with pig densities of 500 to 1,000 pig/km and farms located at a range within 0.5 km and 0.7 km from major roads were more likely to be infected by PRRSV, whereas farms at an elevation of 41 to 61 meters and surrounded by denser vegetation were less likely to be infected, indicating their role as dissemination barriers. In conclusion, our results demonstrate that external dissemination was intense, and reinforce the importance of farm proximity on PRRSV spread. Thus, consideration of farm location, geographic characteristics and animal densities across production systems may help to forecast PRRSV collateral dissemination.
Emergence of novel recombination lineage 3 of porcine reproductive and respiratory syndrome viruses in Southern China.
Sun Yan-Kuo,Li Qi,Yu Zhi-Qing,Han Xiao-Liang,Wei Ying-Fang,Ji Chi-Hai,Lu Gang,Ma Chun-Quan,Zhang Gui-Hong,Wang Heng
Transboundary and emerging diseases
Lineage 3 of porcine reproductive and respiratory syndrome viruses, which belong to North America type 2, has a long epidemic history in China. The novel lineage 3 viruses constantly emerging in recent years are characterized by a high detection rate and significant pathogenicity. In this study, we investigated the prevalence of lineage 3 in southern China and selected two isolated strains for genome and virulence analyses. A cross-sectional epidemiology investigation indicated that the prevalence of lineage 3 antigens was 35.68% (95% CI: 27.6-44.3%) among 227 samples collected from over 100 infected farms from January 2016 to July 2017 in southern China. Two novel isolates of lineage 3 were selected. After 20 passages, Marc-145 cells were not susceptible to those viruses. Full-length genome analysis indicated that the two strains share 95.2% homology with each other and 95.7%-96.2% with highly pathogenic porcine reproductive and respiratory syndrome viruses (HP-PRRSVs; JXA1-like strain, lineage 8.7). Phylogenetic and molecular evolutionary results showed that for the two isolates, HP-PRRSV provides most of the ORF1 gene. Animal experiment revealed discrepancies in virulence between the strains. Although challenge resulted in 100% morbidity, the isolate carrying most of the HP-PRRSV ORF1 caused severe clinical symptoms and 40% mortality, whereas the other isolate containing part of the ORF1 gene caused no mortality. Overall, these findings suggest that lineage 3 viruses might be commonly circulating in most of southern China. Frequent recombination events within HP-PRRSVs of this lineage with changing virulence could represent potential threats to the pig industry.
Characterization of four types of MLV-derived porcine reproductive and respiratory syndrome viruses isolated in unvaccinated pigs from 2016 to 2020.
Chen Nanhua,Li Xinshuai,Xiao Yanzhao,Li Shubin,Zhu Jianzhong
Research in veterinary science
Modified live vaccines (MLVs) have been utilized to combat porcine reproductive and respiratory syndrome (PRRS), which raises a serious concern about the MLV-derived PRRS virus (PRRSV) isolates. During the routine investigation of PRRSV in China, four lung samples collected from unvaccinated diseased pigs from 2016 to 2020 were detected as PRRSV positive. The PRRSVs shared high ORF5 identities to CH-1R, JXA1-R, TJM-F92 and RespPRRS MLV vaccines, respectively. The viruses were isolated in Marc-145 cells and denominated as SD1612-1, JS1703-21, JSTZ1907-714 and JSYC20-05-1. Genome comparison confirmed that these isolates share the highest genomic homologies to CH-1R (97.96%), JXA1-R (99.64%), TJM-F92 (99.00%) and RespPRRS MLV (99.57%) than any other known isolates. Genome-based phylogenetic analysis showed that SD1612-1 and CH-1R, JS1703-21 and JXA1-R, JSTZ1907-714 and TJM-F92, JSYC20-05-1 and RespPRRS MLV were grouped in the same branches. In addition, amino acids unique to corresponding vaccine attenuations were also identified in our isolates. Noticeably, amino-acids potentially associated with the virulence revision from MLV strains to parental virulent viruses were also identified in the MLV-derived isolates. Our results confirm that the four types of MLV-derived isolates are circulating and evolving in Chinese swine herds for years, which highlights the necessity for the fair use of PRRS MLVs.
Histologic Changes Associated With Placental Separation in Gilts Infected with Porcine Reproductive and Respiratory Syndrome Virus.
Novakovic Predrag,Detmer Susan E,Suleman Muhammad,Malgarin Carol M,MacPhee Daniel J,Harding John C S
The placenta is a vital organ providing the developing fetus with nutrient and gas exchange, thermoregulation, and waste elimination necessary for fetal development, as well as producing hormones to maintain pregnancy. It is hypothesized that fetal pig death in porcine reproductive and respiratory syndrome may be attributed to pathology of the maternal-fetal interface leading to premature placental separation. This study was designed to evaluate the chronologic progression of porcine reproductive and respiratory syndrome virus (PRRSV)-induced lesions at the maternal-fetal interface, with particular focus on placental separation in experimentally challenged third-trimester gilts. Fifteen gilts were inoculated with a virulent strain of PRRSV-2 on gestation day 86 ± 0.4. On multiple days postinoculation, 3 gilts along with 1 sham-inoculated control per time point were euthanized, and uterine and fetal placental tissues corresponding to each fetus were collected for histopathologic evaluation. The presence of any fetal lesion was 23 times more likely in compromised (meconium-stained and decomposed) compared with viable fetuses ( P < .001). In PRRSV-infected gilts, endometritis was more severe than placentitis, and the severity of endometrial inflammation and vasculitis increased progressively from 2 to 14 days postinoculation. Neither placental vasculitis nor a chronologic progression in the severity of placental detachment was observed. Severe placental detachment was more frequently present in PRRSV-infected compared with noninfected samples and was most significantly associated with placental inflammation, compared with other uterine lesions, viral load, or termination day. The results of this study suggest that placental separation by itself is not sufficient to significantly compromise fetal viability in reproductive porcine reproductive and respiratory syndrome.
A retrospective study of porcine reproductive and respiratory syndrome virus infection in Brazilian pigs from 2008 to 2020.
Gava Danielle,Caron Luizinho,Schaefer Rejane,Silva Virgínia Santiago,Weiblen Rudi,Flores Eduardo Furtado,de Lima Marcelo,Takeda Guilherme Zaha,Ciacci-Zanella Janice Reis
Transboundary and emerging diseases
Porcine reproductive and respiratory syndrome (PRRS) is a viral disease characterized by reproductive impairment or failure in breeding animals, and a respiratory disease in pigs of any age. Brazil is the fourth largest pork producer and exporter globally, and PRRS virus (PRRSV) infection has never been reported in the country. This study aimed to investigate the status of porcine biological samples from commercial swine herds, quarantined imported boars, wild boars and feral pigs to update PRRS information in Brazil. A total of 14,382 samples were collected from 2008 to 2020, including sera (n = 12,841), plasma (n = 1,000) and oral fluids (n = 541), comprehending 137 herds and free-living pigs in eight Brazilian states. One out of 1,000 (0.1%) plasma and 15 out of 12,841 (0.11%) serum samples tested positive for PRRSV antibodies through ELISA. Upon ELISA retesting, only the plasma sample, from one 8-day-old piglet remained positive. All sixteen previously PRRSV antibody-positive samples were tested through RT-PCR and found to be negative. The presence of false-positive or singleton reactors are quite expected. Thus, the use of different/alternative diagnostic tests is indicated for an efficient PRRSV detection. Taken together, our findings demonstrated no conclusive evidence of PRRSV infection in the tested pigs, highlighting the importance to reinforce the surveillance program to prevent the introduction and eventual dissemination of PRRSV in Brazil.
Porcine Reproductive and Respiratory Syndrome Virus Infection Upregulates Negative Immune Regulators and T-Cell Exhaustion Markers.
Journal of virology
Porcine alveolar macrophage (PAM) is one of the primary cellular targets for porcine reproductive and respiratory syndrome virus (PRRSV), but less than 2% of PAMs are infected with the virus during the acute stage of infection. To comparatively analyze the host transcriptional response between PRRSV-infected PAMs and bystander PAMs that remained uninfected but were exposed to the inflammatory milieu of an infected lung, pigs were infected with a PRRSV strain expressing green fluorescent protein (PRRSV-GFP), and GFP (PRRSV infected) and GFP (bystander) cells were sorted for RNA sequencing (RNA-seq). Approximately 4.2% of RNA reads from GFP and 0.06% reads from GFP PAMs mapped to the PRRSV genome, indicating that PRRSV-infected PAMs were effectively separated from bystander PAMs. Further analysis revealed that inflammatory cytokines, interferon-stimulated genes, and antiviral genes were highly upregulated in GFP compared to GFP PAMs. Importantly, negative immune regulators, including NF-κB inhibitors (NFKBIA, NFKBID, NFKBIZ, and TNFAIP3) and T-cell exhaustion markers (programmed death ligand-1 [PD-L1], PD-L2, interleukin-10 [IL-10], IDO1, and transforming growth factor β2 [TGFB2]) were highly upregulated in GFP cells compared to GFP cells. By using an hybridization assay, RNA transcripts of tumor necrosis factor (TNF) and NF-κB inhibitors were detected in PRRSV-infected PAMs cultured and lung sections of PRRSV-infected pigs during the acute stage of infection. Collectively, the results suggest that PRRSV infection upregulates expression of negative immune regulators and T-cell exhaustion markers in PAMs to modulate the host immune response. Our findings provide further insight into PRRSV immunopathogenesis. Porcine reproductive and respiratory syndrome virus (PRRSV) is widespread in many swine-producing countries, causing substantial economic losses to the swine industry. Porcine alveolar macrophage (PAM) is considered the primary target for PRRSV replication in pigs. However, less than 2% of PAMs from acutely infected pigs are infected with the virus. In the present study, we utilized a PRRSV strain expressing green fluorescent protein to infect pigs and sorted infected and bystander PAMs from the pigs during the acute stage of infection for transcriptome analysis. PRRSV-infected PAMs showed a distinctive gene expression profile and contained many uniquely activated pathways compared to bystander PAMs. Interestingly, upregulated expression of NF-κB signaling inhibitors and T-cell exhaustion molecules were observed in PRRSV-infected PAMs. Our findings provide additional knowledge on the mechanisms that PRRSV employs to modulate the host immune system.
The glycosyltransferase ST3GAL2 modulates virus proliferation and the inflammation response in porcine reproductive and respiratory syndrome virus infection.
Li Xiaoyang,Guo Yanyu,Song Yinna,Sun Ruiqi,Zhu Min,Tan Zheng,Swaiba Umm E,Zhang Lilin,Huang Jinhai
Archives of virology
β-galactoside α-2,3-sialyltransferase 2 (ST3GAL2) is a member of the sialyltransferase family that mediates terminal modification of glycoproteins and glycolipids. ST3GAL2 has been found to play a role in obesity, aging, and malignant diseases. In this study, we cloned porcine ST3GAL2 (pST3GAL2) from porcine alveolar macrophages (PAMs), and its role in porcine reproductive and respiratory syndrome virus (PRRSV) infection was investigated by transcriptome analysis. pST3GAL2 was found to be located in the Golgi apparatus, and it was expressed at high levels in PRRSV-infected PAMs. Overexpression of pST3GAL2 resulted in a slight increase in PRRSV proliferation, and the interaction between pST3GAL2 and GP2a of PRRSV was detected by coimmunoprecipitation and confocal microscopy. The expression of pro-inflammatory cytokines (IFN-β, IL-2, IL-6, IL-18, IL-1β and TNF-α) was significantly inhibited in pST3GAL2-overexpressing, PRRSV-infected cells and upregulated in PRRSV-infected pST3GAL2-knockout cells, while the pattern of expression of anti-inflammatory cytokines (IL-4 and IL-10) was diametrically opposite. Our results demonstrate that the regulation of pST3GAL2 plays an important role in PRRSV proliferation and functional alterations in virus-infected cells. These results contribute to our understanding of the role of β-galactoside α-2,3-sialyltransferase 2 in antiviral immunity.
Evolutionary and recombination analysis of porcine reproductive and respiratory syndrome isolates in China.
Zhang Zhendong,Qu Xiangyang,Zhang Hongliang,Tang Xudong,Bian Ting,Sun Yingjun,Zhou Mingming,Ren Fubo,Wu Ping
Seven strains of porcine reproductive and respiratory syndrome virus (PRRSV) were isolated from 2014 to 2017 in the Shandong province of China and their genomes were sequenced and analyzed. Results showed that all seven of the isolates belong to PRRSV 2, and are clustered into four lineages (lineage 1, 3, 5 and 8) based on comparisons of the ORF5 gene. Comparative analysis of genomes and specific amino acid sites revealed that three of the strains (SDwh1402, SDwh1602 and SDwh1701) have evolved directly from modified live virus (MLV) JXA1-P80, TJM-F92 and IngelvacPRRS. Further recombination analysis revealed that two of the strains (SDyt1401 and SDwh1601) were the result of a recombination event between MLVs JXA1-P80 and NADC30 while two other strains (SDwh1403 and SDqd1501) were the result of recombination between MLVs IngelvacPRRS and NADC30 and HP-PRRSV and QYYZ, respectively. Our results add to the data on MLV evolution and PRRSV recombination and provide a better understanding of the epidemiology of PRRSV in China.
A Molecular and Epidemiological Description of a Severe Porcine Reproductive and Respiratory Syndrome Outbreak in a Commercial Swine Production System in Russia.
Havas Karyn A,Makau Dennis N,Shapovalov Sergei,Tolkova Ekaterina,VanderWaal Kim,Tkachyk Tymofii,Spronk Gordon D,Heron Brad,Dee Scott A,Perez Andres
Porcine reproductive and respiratory syndrome (PRRS) is an economically devastating disease of swine in many parts of the world. Porcine reproductive and respiratory syndrome virus (PRRSV) type 1 is endemic in Europe, and prevalence of the subtypes differ spatially. In this study, we investigated a severe PRRS outbreak reported in 30 farms located in eastern Russia that belong to a large swine production company in the region that was also experiencing a pseudorabies outbreak in the system. Data included 28 ORF5 sequences from samples across 18 of the 25 infected sites, reverse transcriptase real-time polymerase chain reaction (RT-qPCR) results from diagnostic testing, reports of clinical signs, and animal movement records. We observed that the outbreak was due to two distinct variants of wildtype PRRSV type 1 subtype 1 with an average genetic distance of 15%. Results suggest that the wildtype PRRSV variants were introduced into the region around 2019, before affecting this production system (i.e., sow farms, nurseries, and finisher farms). Clinical signs did not differ between the variants, but they did differ by stage of pig production. Biosecurity lapses, including movement of animals from infected farms contributed to disease spread.
Cytotoxic T lymphocyte epitopes identified from a contemporary strain of porcine reproductive and respiratory syndrome virus enhance CD4+CD8+ T, CD8+ T, and γδ T cell responses.
Cao Qian M,Tian Debin,Heffron C Lynn,Subramaniam Sakthivel,Opriessnig Tanja,Foss Dennis L,Calvert Jay G,Meng Xiang-Jin
Immuno-stimulatory class I-restricted cytotoxic T lymphocytes (CTL) epitopes of porcine reproductive and respiratory syndrome virus (PRRSV) are important for vaccine development. In this study we first determined the expression frequency of swine leukocyte antigen (SLA) class I alleles in commercial pigs in the United States. The SLA genotyping result allowed us to predict potential CTL epitopes from a contemporary strain of PRRSV (RFLP 1-7-4) by using bioinformatic tools. The predicted epitopes were then evaluated in an ex vivo stimulation assay with peripheral blood mononuclear cells isolated from pigs experimentally-infected with PRRSV. Using flow-cytometry analysis, we identified a number of immuno-stimulatory CTL epitopes, including two peptides from GP3 and two from Nsp9 that significantly improved both degranulation marker CD107a and IFN-γ production in cytotoxic CD4+CD8+ T cells, CD8+ T cells, and γδ T cells, and two peptides that inhibited IFN-γ production. These CTL epitopes will aid future vaccine development against PRRSV.
The prevalent status and genetic diversity of porcine reproductive and respiratory syndrome virus in China: a molecular epidemiological perspective.
Guo Zhenhua,Chen Xin-Xin,Li Rui,Qiao Songlin,Zhang Gaiping
Porcine reproductive and respiratory syndrome virus (PRRSV) has been epidemic more than 30 years in America and 20 years in China. It is still one of the most important causative agents to the worldwide swine industry. Here, we systematically analyzed the prevalence status of PRRSV in China by a molecular epidemiological perspective. Now both PRRSV-1 and PRRSV-2 are circulating and approximately more than 80% of pig farms are seropositive for PRRSV. For PRRSV-2, there are four lineages (lineage 1, lineage 3, lineage 5, lineage 8) circulating in the fields. Lineage 8 (CH-1a-like) and lineage 5 (BJ-4-like) appeared almost at the same time during 1995-1996. Notably, BJ-4 shares 99.6% and 99.8% identity with VR2332 and RespPRRS MLV, respectively. It means that lineage 5 is likely to be imported from America. Now highly pathogenic PRRSV (HP-PRRSV) which was considered to be evolved from local diversity of lineage 8 strains is predominant with different variants. Lineage 3 appeared in 2010 which is mainly sporadic in south of China. Lineage 1, also known as NADC30-like strains in China, has been prevalent since 2013 and leads to PRRS pandemic again. For PRRSV-1, although sporadic at present, more than 9 provinces/regions have been reported. All the circulating strains belong to subtype I. It should be paid more attention since there are no vaccines available. Our analysis would help to deeply understand the prevalent status of PRRSV in China and provide useful information for prevention and control of porcine reproductive and respiratory syndrome (PRRS).
Genetic characterization of a novel recombined porcine reproductive and respiratory syndrome virus 2 among Nadc30-like, Jxa1-like and TJ-like strains.
Zhao Jun,Zhu Ling,Huang Jianbo,Yang Zexiao,Xu Lei,Gu Sirui,Huang Yao,Zhang Rubo,Sun Xiangang,Zhou Yuancheng,Xu Zhiwen
Veterinary medicine and science
Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating viral diseases in the global pig industry, including China. Recently, we successfully isolated a porcine reproductive and respiratory syndrome virus (PRRSV) from lung tissue and peripheral blood of piglets at a farm from Dujiangyan in Sichuan, China, and named it the DJY-19 strain. The full-length genome sequence of DJY-19 shared 86.8%-94.1% nucleotide similarity with NADC30-like and NADC30 PRRSV strains. We compared the open reading frame (ORF) 5 gene of DJY-19 with 34 PRRSV strains from Genbank. Phylogenetic analysis showed that DJY-19 clustered with NADC30 strains, characterized by a predicted 131-amino-acid deletion in the nonstructural protein (NSP) 2. The results of homology analysis showed that the homology between DJY-19 and NADC30 (JN654459.1) strains was the highest (95.9%), whereas homology with other domestic strains was lower (80.9%-92.6%). Furthermore, we identified four recombination breakpoints in the DJY-19 genome; they separated the DJY-19 genome into four regions. The 8106-9128 nucleotide (nt) region of DIY-19 was highly similar to the TJ strain, and the 12106-12580 nt region of DIY-19 was highly similar to the JXA1-R strain. Our findings demonstrate that DJY-19 arose from the recombination of North America NADC30 strain and TJ strain and JXA1-R in China. The application of multiple attenuated vaccine strains has led to complex recombination of PRRSV strains in China. This study provides a theoretical basis for making a more reasonable PRRS virus control and prevention strategy.
Nsp2 and GP5-M of Porcine Reproductive and Respiratory Syndrome Virus Contribute to Targets for Neutralizing Antibodies.
Su Jia,Zhou Lei,He Bicheng,Zhang Xinhui,Ge Xinna,Han Jun,Guo Xin,Yang Hanchun
Porcine reproductive and respiratory syndrome virus (PRRSV) is characterized by its genetic variation and limited cross protection among heterologous strains. Even though several viral structural proteins have been regarded as inducers of neutralizing antibodies (NAs) against PRRSV, the mechanism underlying limited cross-neutralization among heterologous strains is still controversial. In the present study, examinations of NA cross reaction between a highly pathogenic PRRSV (HP-PRRSV) strain, JXwn06, and a low pathogenic PRRSV (LP-PRRSV) strain, HB-1/3.9, were conducted with viral neutralization assays in MARC-145 cells. None of the JXwn06-hyperimmuned pigs' sera could neutralize HB-1/3.9 in vitro and vice versa. To address the genetic variation between these two viruses that are associated with limited cross-neutralization, chimeric viruses with coding regions swapped between these two strains were constructed. Viral neutralization assays indicated that variations in nonstructural protein 2 (nsp2) and structural proteins together contribute to weak cross-neutralization activity between JXwn06 and HB-1/3.9. Furthermore, we substituted the nsp2-, glycoprotein2 (GP2)-, GP3-, and GP4-coding regions together, or nsp2-, GP5-, and membrane (M) protein-coding regions simultaneously between these two viruses to construct chimeric viruses to test cross-neutralization reactivity with hyperimmunized sera induced by their parental viruses. The results indicated that the swapped nsp2 and GP5-M viruses increased the neutralization reactivity with the donor strain antisera in MARC-145 cells. Taken together, these results show that variations in nsp2 and GP5-M correlate with the limited neutralization reactivity between the heterologous strains HP-PRRSV JXwn06 and LP-PRRSV HB-1/3.9.
Genomic sequence and virulence of a novel NADC30-like porcine reproductive and respiratory syndrome virus isolate from the Hebei province of China.
Sui Xiukun,Guo Xiaoyu,Jia Hong,Wang Xixi,Lin Weidong,Li Ming,Gao Xintao,Wu Jing,Jiang Yitong,Willems L,Zhu Hongfei,Xin Ting,Hou Shaohua
Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome (PRRS), which results in immense economic losses in the swine industry. Outbreaks of disease caused by NADC30-like PRRSV are of great concern in China. Here, a novel variant, NADC30-like PRRSV strain HB17A, was analyzed and its pathogenicity in pigs was examined. The full-length genome sequence of HB17A shared 83.6-95.1% nucleotide similarity with NADC30-like and NADC30 PRRSV without any gene insertions, but with a unique 2-amino acid deletion in Nsp2. A phylogenetic analysis showed that HB17A clustered with NADC30 strains. Different degrees of variation in the signal peptide, transmembrane region (TM), primary neutralizing epitope (PNE), non-neutral epitopes, and N-glycosylation sites were observed in GP5. Challenge experiments showed that HB17A infection resulted in persistent fever, moderate respiratory clinical signs, low levels of viremia and viral loads in serum, and mild gross and microscopic lung lesions. Moreover, IFN-γ, IL-6, and IL-10 cytokine levels were significantly elevated in serum, but the levels of IFN-α and IL-2 were similar to those of the negative controls. HB17A was less pathogenic but was secreted longer in nasal discharge than HP-PRRSV FZ06A. Our findings indicate that HB17A is a novel NADC30-like strain with certain deletions and mutations but with no evidence of genomic recombination. This strain exhibits intermediate virulence in pigs. This research will be help define the evolutionary characteristics of Chinese NADC30-like PRRSV.
Insights into the evolutionary history and epidemiological characteristics of the emerging lineage 1 porcine reproductive and respiratory syndrome viruses in China.
Sun Yan-Kuo,Chen Yong-Jie,Cai Yu,Li Qi,Xie Jie-Xiong,Liang Guan,Gao Qi,Yu Zhi-Qing,Lu Gang,Huang Liang-Zong,Ma Chun-Quan,Gong Lang,Wang Heng,Shi Mang,Zhang Gui-Hong
Transboundary and emerging diseases
The newly emerged lineage 1 porcine reproductive and respiratory syndrome viruses (PRRSVs) (especially the NADC30-like and NADC34-like viruses) have posed a direct threat to the Chinese pig industry since 2013. The phylogenetic, epidemic, and recombinant properties of these viruses have not yet systematically analysed in China. This report presents regular surveillance and field epidemiological studies for PRRSV across China from 2007 to 2019. From over 4,000 detected clinical samples, 70 open reading frame five sequences and four complete genomes of lineage 1 viruses were successfully obtained. Combined with global data, we conducted an extensive and systematic molecular phylogeny analysis using a maximum likelihood tree. The Chinese lineage 1 viruses were clustered, and their temporal and spatial distribution was further explored. Multiple viral introductions of lineage 1 virus from the United States to China were detected, and some became endemic in China. There are three sub-lineage 1 clusters: lineage 1.5 (NADC34-like), lineage 1.6 and New Intro cluster (NADC30-like). These viruses show high genetic diversity and a wide distribution in China, with Henan Province showing the highest diversity. Moreover, Chinese lineage 1 viruses have developed an endemic NADC30-like cluster. The demographic feature of this cluster showed a more or less constant population expansion history with a recent decreasing trend. Moreover, the genome recombination of Chinese lineage 1 with two dominant clusters (Chinese HP-PRRSVs: lineage 8.7 and VR2332-like: lineage 5.1) was frequently detected, both of which have commercial vaccine strains available. Furthermore, recombination hotspots were discovered near NSP9 and ORF2-4 regions of the genome. Overall, these findings provide important insights into the evolution and geographical diversity of Chinese lineage 1 PRRSV. These results will facilitate the development of programmes for the control and prevention of the emerging lineage 1 viruses in China.
Two immune-based methods using immortalized porcine kidney macrophages for quantifying neutralizing activity against porcine reproductive and respiratory syndrome virus-2.
Terada Takumi,Morozumi Takeya,Wada Emi,Sukegawa Shin
Journal of virological methods
Porcine reproductive and respiratory syndrome virus (PRRSV) causes a serious infectious disease in pigs in farms worldwide. Neutralizing antibody titer is an effective index for evaluating immunity to PRRSV; however, PRRSV has different neutralizing cross-reactivity between strains. Therefore, quantitative measurement of neutralizing antibody titers against field PRRSV strains would be required to evaluate whether neutralizing antibodies in pigs could possess neutralizing activity against individual or multiple strains. Immune-based methods, such as image cytometry (ICM) and cell-based enzyme-linked immune sorbent assay (ELISA), are quantitative and can be used to evaluate many samples. Using immortalized porcine kidney macrophages (IPKMs), which are highly susceptible to infection from field PRRSV-2 strains compared with other cell lines, immune-based methods could enable the evaluation of the neutralizing activity of porcine serum against field strains of PRRSV-2 that are difficult to isolate in conventional cells. In summary, we adapted two methods, namely ICM and cell-based ELISA, to IPKMs for quantitative neutralizing antibody titer measurements. Two immune-based methods using IPKMs are adequate for quantifying neutralizing activity of porcine serum against PRRSV-2, including field strains.
Co-infection status of classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCV2 and PCV3) in eight regions of China from 2016 to 2018.
Chen Nanhua,Huang Yucheng,Ye Mengxue,Li Shuai,Xiao Yanzhao,Cui Bailei,Zhu Jianzhong
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases
Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circoviruses (PCV2 and PCV3) are economically important swine viruses that cause reproductive failure and/or respiratory symptoms in pigs. However, the co-infection status of these viruses in Chinese swine herds is not well clarified. In this study, we evaluated the co-infection of these four viruses in 159 pigs collected from 63 herds in eight regions of China from 2016 to 2018. CSFV, PRRSV, PCV2 and PCV3 were detected in 14, 56, 43 and 4 of the pigs, respectively. The percentage of singular infections was 32.71%, while the percentages of dual infections and multiple infections were 15.72% and 3.15%, respectively. The E2 of CSFV, ORF5 of PRRSV, ORF2s of PCV2 and PCV3 from all positive samples were determined and used for phylogenetic analyses. E2-based phylogenetic tree showed that all 14 CSFVs identified in this study belong to 2.1b subtype. ORF5-based phylogenetic tree showed that PRRSV2 is predominant in China while PRRSV1 can also be detected. In addition, 35, 16, 4 and 1 of our PRRSVs are clustered with highly pathogenic PRRSV2, NADC30-like PRRSV2, classical PRRSV2 and PRRSV1, respectively. ORF2-based phylogenetic trees showed that our PCVs are grouped with 2 PCV2 subtypes (PCV2d and PCV2b) and 3 PCV3 subtypes (PCV3a, PCV3b and PCV3c), respectively. Our results provide the latest co-infection status and the diversity of four important swine viruses in Chinese swine herds, which is beneficial for understanding the epidemiology of these viruses.
Identification of Virulence Associated Region during Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus during Attenuation In Vitro: Complex Question with Different Strain Backgrounds.
Jiang Yifeng,Tong Wu,Yu Lingxue,Li Liwei,Gao Fei,Li Guoxin,Liu Changlong,Chen Pengfei,Shen Qi,Zhang Yujiao,Zhou Yanjun,Tong Guangzhi
Highly pathogenic porcine reproductive and respiratory syndrome virus PRRSV (HP-PRRSV) was one of the most devastating diseases of the pig industry, among various strategies, vaccination was one of the most useful tools for PRRS control. Attenuated live vaccine was used worldwide, however, the genetic basis of HP-PRRSV virulence change during attenuation remain to be determined. Here, to identify virulence associated regions of HP-PRRSV during attenuation in vitro, six full-length infectious cDNA clones with interchanges of 5'UTR + ORF1a, ORF1b, and ORF2-7 + 3'UTR regions between HP-PRRSV strain HuN4-F5 and its attenuated vaccine strain HuN4-F112 were generated, and chimeric viruses were rescued. Piglets were inoculated with chimeric viruses and their parental viruses, and rectal temperature were recorded daily, and serum were collected for future experiments. Our results showed that ORF1a played an important role on virus replication, cytokine response and lung damage, the exchange of ORF1b and ORF2-7 in different backbone led to different exhibition on virus replication in vivo/vitro and cytokine response. Among 9 PRRSV attenuated series, consistent amino acid changes during PRRSV attenuation were found in NSP4, NSP9, GP2, E, GP3 and GP4. Our study provides a fundamental data for the investigation of PRRSV attenuation, the different results of the virulence change among different studies indicated that different mechanisms might be used during PRRSV virulence enhancement in vivo and attenuation in vitro.
Generation and Evaluation of Recombinant Baculovirus Coexpressing and Proteins of Porcine Reproductive and Respiratory Syndrome Virus Type 1.
Xu Wang,Du Shouwen,Li Tiyuan,Wu Shipin,Jin Ningyi,Ren Linzhu,Li Chang
Porcine reproductive and respiratory syndrome virus (PRRSV) is the pathogen of the porcine reproductive and respiratory syndrome, which is one of the most economically devastating diseases of the swine industry. However, whether the inactivated vaccine and modified live attenuated vaccines are effective in disease control is still controversial. Although several groups developed PRRSV virus-like particles (VLPs) as a vaccine against PRRSV, all these VLP-based vaccines targeted PRRSV-2, but not PRRSV-1 or both. Therefore, it is urgent to produce VLPs against PRRSV-1. In this study, we rescued recombinant baculovirus expressing and proteins of PRRSV-1 through the Bac-to-Bac baculovirus expression system. Thereafter, PRRSV VLP was obtained efficiently in the recombinant baculovirus-infected High Five insect cells. Moreover, the PRRSV VLP and PRRSV VLP+A5 could efficiently trigger specific humoral immune responses and B cellular immune responses through intranasal immunization. The combination of PRRSV VLP and A5 adjuvant could improve the level of the immune response. The PRRSV-1 VLPs generated in this study have greater potential for vaccine development to control PRRSV-1 infection.
Genomic characteristics and pathogenicity of natural recombinant porcine reproductive and respiratory syndrome virus 2 harboring genes of a Korean field strain and VR-2332-like strain.
Kwon Taeyong,Yoo Sung J,Park Jun Woo,Kang Sang Chul,Park Choi-Kyu,Lyoo Young S
Porcine reproductive and respiratory syndrome (PRRS), an economically-important disease caused by PRRS virus (PRRSV), has become endemic to most pig-producing countries. Point mutation and recombination are responsible for genetic heterogeneity, resulting in circulation of genetically-diverse strains. However, no natural recombinant PRRSV has yet been identified in Korea. Here, we successfully isolated natural recombinant PRRSV-2 (KU-N1202) using cell culture, investigated its genomic characteristics, and further evaluated its pathogenicity. KU-N1202 is a recombinant strain between Korean MN184-like and VR-2332-like strains. Specifically, ORF5 to partial ORF7 of the VR-2332-like strain was inserted into the backbone of a CP07-626-2-like strain. KU-N1202 induced mild-to-moderate clinical signs and mild histopathological changes with low viral loads in challenged pigs. Contact pigs showed minimal clinical signs and lower viral loads than those in the challenge group. This study demonstrates the genomic characteristics and pathogenicity of natural recombinant PRRSV-2, illustrating the potential importance of recombination in the field.
Immunization with a recombinant fusion of porcine reproductive and respiratory syndrome virus modified GP5 and ferritin elicits enhanced protective immunity in pigs.
Ma Hui,Li Xiangmin,Li Jianglong,Zhao Zekai,Zhang Huawei,Hao Genxi,Chen Huanchun,Qian Ping
Porcine reproductive and respiratory syndrome (PRRS) has caused huge economic losses in the swine industry worldwide. Live and inactivated vaccines have only been partially successful in generating protective immune responses. The PRRS virus (PRRSV) glycoprotein 5 (GP5) is a major viral antigenic target and is thus suitable for development of genetically engineered PRRSV vaccines. Here, a modified GP5 and ferritin were fused and expressed using a baculovirus system to generate a GP5m-ferritin nanoparticle vaccine. We demonstrated that the GP5m-ferritin vaccine elicited higher serum antibody titers in pigs than inactivated PRRSV. Moreover, immunization with GP5m-Ft promoted a Th1-dominant cellular immune response and enhanced specific T-lymphocyte immune responses. GP5m-ferritin-vaccinated pigs had significantly lower mean rectal temperatures, respiratory scores, viremia, and macroscopic and microscopic lung lesion scores post-challenge compared with unvaccinated pigs. These results indicated that GP5m-ferritin subunit vaccines can elicit specific protective immune responses and represent promising vaccine candidates.
Porcine Reproductive and Respiratory Syndrome (PRRS) Epidemiology in an Integrated Pig Company of Northern Italy: A Multilevel Threat Requiring Multilevel Interventions.
Franzo Giovanni,Barbierato Giacomo,Pesente Patrizia,Legnardi Matteo,Tucciarone Claudia Maria,Sandri Giampietro,Drigo Michele
Porcine reproductive and respiratory syndrome (PRRS) is probably the most relevant viral disease affecting pig farming. Despite the remarkable efforts paid in terms of vaccination administration and biosecurity, eradication and long-term control have often been frustrated. Unfortunately, few studies are currently available that objectively link, using a formal statistical approach, viral molecular epidemiology to the risk factors determining the observed scenario. The purpose of the present study is to contribute to filling this knowledge gap taking advantage of the advancements in the field of phylodynamics. Approximately one-thousand ORF7 sequences were obtained from strains collected between 2004 and 2021 from the largest Italian pig company, which implements strict compartmentalization among independent three-sites (i.e., sow herds, nurseries and finishing units) pig flows. The history and dynamics of the viral population and its evolution over time were reconstructed and linked to managerial choices. The viral fluxes within and among independent pig flows were evaluated, and the contribution of other integrated pig companies and rurally risen pigs in mediating such spreading was investigated. Moreover, viral circulation in Northern Italy was reconstructed using a continuous phylogeographic approach, and the impact of several environmental features on PRRSV strain persistence and spreading velocity was assessed. The results demonstrate that PRRSV epidemiology is shaped by a multitude of factors, including pig herd management (e.g., immunization strategy), implementation of strict-independent pig flows, and environmental features (e.g., climate, altitude, pig density, road density, etc.) among the others. Small farms and rurally raised animals also emerged as a potential threat for larger, integrated companies. These pieces of evidence suggest that none of the implemented measures can be considered effective alone, and a multidimensional approach, ranging from individual herd management to collaboration and information sharing among different companies, is mandatory for effective infection control.
Evolutionary Patterns of Codon Usage in Major Lineages of Porcine Reproductive and Respiratory Syndrome Virus in China.
Wu Weixin,Ge Xinna,Zhang Yongning,Han Jun,Guo Xin,Zhou Lei,Yang Hanchun
Porcine reproductive and respiratory syndrome virus (PRRSV) is economically important and characterized by its extensive variation. The codon usage patterns and their influence on viral evolution and host adaptation among different PRRSV strains remain largely unknown. Here, the codon usage of ORF5 genes from lineages 1, 3, 5, and 8, and MLV strains of type 2 PRRSV in China was analyzed. A compositional property analysis of ORF5 genes revealed that nucleotide C is most frequently used at the third position of codons, accompanied by rich GC3s. The effective number of codon (ENC) and codon pair bias (CPB) values indicate that all ORF5 genes have low codon bias and the differences in CPB scores among four lineages are almost not significant. When compared with host codon usage patterns, lineage 1 strains show higher CAI and SiD values, with a high similarity to pig, which might relate to its predominant epidemic propensity in the field. The CAI, RCDI, and SiD values of ORF5 genes from different passages of MLV JXA1R indicate no relation between attenuation and CPB or codon adaptation decrease during serial passage on non-host cells. These findings provide a novel way of understanding the PRRSV's evolution, related to viral survival, host adaptation, and virulence.
Analysis of ORF5 sequences of Porcine Reproductive and Respiratory Syndrome virus (PRRSV) circulating within swine farms in Costa Rica.
Guzmán Mónica,Meléndez Ronald,Jiménez Carlos,Piche Marta,Jiménez Emily,León Bernal,Cordero Juan M,Ramirez-Carvajal Lisbeth,Uribe Alberto,Van Nes Arie,Stegeman Arjan,Romero Juan José
BMC veterinary research
BACKGROUND:Worldwide, Porcine Reproductive and Respiratory Syndrome (PRRS) is among the diseases that cause the highest economic impact in modern pig production. PRRS was first detected in Costa Rica in 1996 and has since then severely affected the local swine industry. Studies of the molecular characterization of circulating strains, correlation with clinical records, and associations with pathogens associated with Porcine Respiratory Disease Complex (PRDC) have not been done in Costa Rica. RESULTS:Sequencing and phylogenetic analysis of ORF5 proved that PRRSV-2 was the only species detected in all locations analyzed. These sequences were grouped into three clusters. When comparing samples from San Jose, Alejuela, and Puntarenas to historical isolates of the previously described lineages (1 to 9), it has been shown that these were closely related to each other and belonged to Lineage 5, along with the samples from Heredia. Intriguingly, samples from Cartago clustered in a separate clade, phylogenetically related to Lineage 1. Epitope analysis conducted on the GP5 sequence of field isolates from Costa Rica revealed seven peptides with at least 80% amino acid sequence identity with previously described and experimentally validated immunogenic regions. Previously described epitopes A, B, and C, were detected in the Santa Barbara-Heredia isolate. CONCLUSIONS:Our data suggest that the virus has three distinct origins or introductions to the country. Future studies will elucidate how recently introduced vaccines will shape the evolutionary change of circulating field strains.
Novel lineage 1 recombinants of porcine reproductive and respiratory syndrome virus isolated from vaccinated herds: genome sequences and cytokine production profiles.
Park Jonghyun,Choi Subin,Jeon Ji Hyun,Lee Kyung-Won,Lee Changhee
Archives of virology
Porcine reproductive and respiratory syndrome virus (PRRSV) is a widely disseminated, macrophage-tropic arterivirus that exhibits profound genetic and pathogenic heterogeneity. The present study was conducted to determine the complete genome sequences of two novel Korean lineage 1 PRRSV-2 strains, KNU-1901 and KNU-1902, which were isolated from vaccinated pig farms experiencing unusually high morbidity and mortality. Both isolates contained notable discontinuous 423-nucleotide deletions (DELs) within the genes encoding nonstructural protein 2 (nsp2) and GP3 when compared with the prototype strain VR-2332. In particular, the nsp2 DEL viruses had unique quadripartite discontinuous DEL signatures (111-1-19-9) in nsp2; this is an expanded version of the tripartite 111-1-19 DEL previously identified in virulent lineage 1 PRRSV-2 strains. Phylogenetic analysis revealed that both novel nsp2 DEL viruses belong to the Korean clade (KOR C) of lineage 1 isolates based on ORF5 but cluster with lineage KOR A strains based on the nsp2 or complete genome sequence. Recombination detection analysis suggested that both novel isolates are recombinants and may have evolved via natural inter-lineage recombination between circulating KOR A and KOR C strains. Interestingly, compared with the prototype VR-2332 virus, the novel nsp2 DEL variants were less efficient at promoting the expression of immune response genes in porcine alveolar macrophage culture. Taken together, we conclude that KNU-1901 and KNU-1902 are recently evolved recombinant variants of the virulent lineage 1 family that caused the regional severe PRRS outbreaks.
Evolutionary Dynamics of Type 2 Porcine Reproductive and Respiratory Syndrome Virus by Whole-Genome Analysis.
Guo Jiahui,Liu Zimin,Tong Xue,Wang Zixin,Xu Shangen,Chen Qian,Zhou Junwei,Fang Liurong,Wang Dang,Xiao Shaobo
Porcine reproductive and respiratory syndrome virus (PRRSV), an important pathogen in the swine industry, is a genetically highly diverse RNA virus. However, the phylogenetic and genomic recombination properties of this virus are not yet fully understood. In this study, we performed an integrated analysis of all available whole-genome sequences of type 2 PRRSV (n = 901) to reveal its evolutionary dynamics. The results showed that there were three distinct phylogenetic lineages of PRRSV in their distribution patterns. We identified that sublineage 2.7 (L2.7), associated with a NADC30 cluster, had the highest substitution rate and higher viral genetic diversity, and inter-lineage recombination is observed more frequently in L2.7 PRRSV compared to other sublineages. Most inter-lineage recombination events detected are observed between L2.7 PRRSVs (as major parents) and L3.4 (a JXA1-R-related cluster)/L3.7 (a WUH3-related cluster) PRRSVs (as minor parents). Moreover, the recombination hotspots are located in the structural protein gene ORF2 and ORF4, or in the non-structural protein gene nsp7. In addition, a GM2-related cluster, L3.2, shows inconsistent recombination modes compared to those of L2.7, suggesting that it may have undergone extensive and unique recombination in their evolutionary history. We also identified several amino acids under positive selection in GP2, GP4 and GP5, the major glycoproteins of PRRSV, showing the driving force behind adaptive evolution. Taken together, our results provide new insights into the evolutionary dynamics of PPRSV that contribute to our understanding of the critical factors involved in its evolution and guide future efforts to develop effective preventive measures against PRRSV.
Recombinant Porcine Interferon Alpha Enhances Immune Responses to Killed Porcine Reproductive and Respiratory Syndrome Virus Vaccine in Pigs.
Yu Hai-Yang,Qu Ming-Sheng,Zhang Jun-Ling,Gan Lin,Zhao Yu,Shan Xue-Qin,Zhou Wei,Xia Bing-Bing,Chen Jason,Wang Ming-Li,Zhao Jun
In this study, the immunoadjuvant effects of recombinant porcine interferon alpha (rPoIFN) on the killed virus vaccine (KV) of porcine reproductive and respiratory syndrome virus (PRRSV) in pigs were investigated. The experimental pigs were divided into six groups, including normal control group, rPoIFN control group, PRRSV KV control group, KV+40,000 U rPoIFN immunization group, KV+400,000 U rPoIFN immunization group, and KV+4,000,000 U rPoIFN immunization group. The experimental pigs were boosted immunized on the 28th day after the initial immunization, and the heparinized blood and serum samples were collected at different time points of these two immunizations to detect and evaluate the immune responses of pigs after immunization by ELISA assay, neutralization assay, flow cytometry, and so on. The results showed that the proportion of the levels of PRRSV-specific antibodies, neutralizing antibodies, stimulation index, IL-4, IFN-γ, and lymphocytes within the groups immunized with KV+rPoIFN were significantly higher than that group immunized with KV alone. The humoral and cellular immune responses in pigs were markedly enhanced by rPoIFN after the coadministration with KV vaccine. Therefore, we tentatively think that rPoIFN is a potential immune promoter with prospects for future applications in the pig industry.
A strain of highly pathogenic porcine reproductive and respiratory syndrome virus: genomic characterization, pathogenicity, and construction of an infectious full-length cDNA clone.
Wang Shuangyun,Liu Yanling,Yu Linyang,Liang Tairun,Zhang Pengfei,Dong Jianguo,Zhang Leyi,Liang Pengshuai,Wang Lei,Xu Zheng,Song Changxu
Archives of virology
Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which inflicts major economic losses on the global pig farming industry. Based on its similarity to highly pathogenic strains, the GDzj strain isolated in this study was predicted to be highly pathogenic. We therefore analyzed the pathogenicity of this strain experimentally in piglets. All piglets challenged with this virus experienced fever or high fever, loss of appetite, decreased food intake, daily weight loss, shortness of breath, and listlessness, and the necropsy results showed that they had experienced severe interstitial pneumonia. We then used the BAC system to construct a full-length cDNA infectious clone of GDzj, and the rescued virus displayed in vitro proliferation characteristics similar to those of the parental PRRSV strain. In summary, we successfully isolated a highly pathogenic PRRSV strain and constructed a full-length infectious cDNA clone from it, thereby providing an effective reverse genetics platform for further study of viral pathogenesis.
Studies on heterologous protection between Japanese type 1 and type 2 porcine reproductive and respiratory syndrome virus isolates.
Iseki Hiroshi,Kawashima Kenji,Takagi Michihiro,Shibahara Tomoyuki,Mase Masaji
The Journal of veterinary medical science
The objective of the present study was to evaluate the cross-protective immunity between type 1 and type 2 porcine reproductive and respiratory syndrome virus (PRRSV) isolates in growing pigs. Japanese type 1 PRRSV, first isolated from a pig with respiratory disorders in a farm in 2009, exhibits unique genetic characteristics. The pathogenicity of a Japanese standard strain of type 2 PRRSV, EDRD1, in pigs immunized by the type 1 PRRSV isolate, Jpn EU 4-37 was determined by evaluating clinical signs, viremia, antibody response, and pathological lesions. Similarly, we evaluated the pathogenicity of Jpn EU 4-37 in pigs immunized by EDRD1 and compared the cross-protective immunity between these isolates. The EDRD1 challenge after Jpn EU 4-37 inoculation reduced viral clearance and shedding in pigs, compared to those treated with the EDRD1 single infection. On the other hand, the pathogenicity of Jpn EU 4-37 after EDRD1 infection did not differ significantly compared to non-immunized pigs treated with Jpn EU 4-37. Therefore, exposure to Jpn EU 4-37 could not induce enough immunity to reduce the viremia against subsequent infection by type 2 PRRSV. However, the immunity induced by Jpn EU 4-37 infection may play a role in reducing viremia caused by type 2 PRRSV. Moreover, the immunity induced by the EDRD1 and other genetically related viruses, which are broadly distributed in Japan, may not contribute to cross-protection against Jpn EU 4-37 as an emerging virus.
Research progress in the development of porcine reproductive and respiratory syndrome virus as a viral vector for foreign gene expression and delivery.
Dai Guo,Huang Mei,Fung To Sing,Liu Ding Xiang
Expert review of vaccines
INTRODUCTION:Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease of swine characterized by respiratory disorders in growing and finishing pigs and reproductive failure in pregnant sows. PRRSV has been recognized as one of the most economically significant pathogens affecting the global pig industry. AREAS COVERED:Currently, commercially available vaccines, including traditional killed virus (KV) vaccines and modified live virus (MLV) vaccines, are the cardinal approaches to prevent and control porcine reproductive and respiratory syndrome virus (PRRSV) infection. However, the protective efficacy of these vaccines is not satisfactory, resulting in the continuous evolution and recurrent appearance of the virus as well as the emergence of new variants. A safe and effective vaccine against PRRSV is in dire need. Here, we review the research progress in recent years in the development and use of PRRSV as a viral vector to express foreign genes, and their potential application in gene delivery and vaccine development. EXPERT OPINION:The potential of using PRRSV-based vectors to express multiple antigens would be particularly instrumental for the development of a new generation of multivalent vaccines against PRRSV and other porcine viruses.
The Ability of Porcine Reproductive and Respiratory Syndrome Virus Isolates to Induce Broadly Reactive Neutralizing Antibodies Correlates With Protection.
Martínez-Lobo Francisco Javier,Díez-Fuertes Francisco,Simarro Isabel,Castro José M,Prieto Cinta
Frontiers in immunology
Porcine reproductive and respiratory syndrome (PRRS) is considered one of the most relevant diseases of swine. The condition is caused by PRRS virus (PRRSV), an extremely variable virus of the family. Its heterogeneity can be responsible, at least partially, of the poor cross-protection observed between PRRSV isolates. Neutralizing antibodies (NAs), known to play a role in protection, usually poorly recognize heterologous PRRSV isolates, indicating that most NAs are strain-specific. However, some pigs develop broadly reactive NAs able to recognize a wide range of heterologous isolates. The aim of this study was to determine whether PRRSV isolates that induce broadly reactive NAs as determined are able to confer a better protection . For this purpose two experiments were performed. Initially, 40 pigs were immunized with a PRRSV-1 isolate known to induce broadly reactive NAs and 24 additional pigs were used as controls. On day 70 after immunization, the pigs were divided into eight groups composed by five immunized and three control pigs and exposed to one of the eight different heterologous PRRSV isolates used for the challenge. In the second experiment, the same experimental design was followed but the pigs were immunized with a PRRSV-1 isolate, which is known to generate mostly strain-specific NAs. Virological parameters, specifically viremia and the presence of challenge virus in tonsils, were used to determine protection. In the first experiment, sterilizing immunity was obtained in three groups, prevention of viremia was observed in two additional groups, although the challenge virus was detected occasionally in the tonsils of immunized pigs, and partial protection, understood as a reduction in the frequency of viremia compared with controls, was recorded in the remaining three groups. On the contrary, only partial protection was observed in all groups in the second experiment. The results obtained in this study confirm that PRRSV-1 isolates differ in their ability to induce cross-reactive NAs and, although other components of the immune response might have contributed to protection, pigs with cross-reactive NAs at the time of challenge exhibited better protection, indicating that broadly reactive NAs might play a role in protection against heterologous reinfections.
Simultaneous Infection With Porcine Reproductive and Respiratory Syndrome and Influenza Viruses Abrogates Clinical Protection Induced by Live Attenuated Porcine Reproductive and Respiratory Syndrome Vaccination.
Chrun Tiphany,Maze Emmanuel A,Vatzia Eleni,Martini Veronica,Paudyal Basudev,Edmans Matthew D,McNee Adam,Manjegowda Tanuja,Salguero Francisco J,Wanasen Nanchaya,Koonpaew Surapong,Graham Simon P,Tchilian Elma
Frontiers in immunology
The porcine respiratory disease complex (PRDC) is responsible for significant economic losses in the pig industry worldwide. Porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus are major viral contributors to PRDC. Vaccines are cost-effective measures for controlling PRRS, however, their efficacy in the context of co-infections has been poorly investigated. In this study, we aimed to determine the effect of PRRSV-2 and swine influenza H3N2 virus co-infection on the efficacy of PRRSV modified live virus (MLV) vaccination, which is widely used in the field. Following simultaneous challenge with contemporary PRRSV-2 and H3N2 field isolates, we found that the protective effect of PRRS MLV vaccination on clinical disease and pathology was abrogated, although viral load was unaffected and antibody responses were enhanced. In contrast, co-infection in non-immunized animals reduced PRRSV-2 viremia and H3N2 virus load in the upper respiratory tract and potentiated T cell responses against both PRRSV-2 and H3N2 in the lung. Further analysis suggested that an upregulation of inhibitory cytokines gene expression in the lungs of vaccinated pigs may have influenced responses to H3N2 and PRRSV-2. These findings provide important insights into the effect of viral co-infections on PRRS vaccine efficacy that may help identify more effective vaccination strategies against PRDC in the field.
Immunity against a Japanese local strain of porcine reproductive and respiratory syndrome virus decreases viremia and symptoms of a highly pathogenic strain.
Iseki Hiroshi,Kawashima Kenji,Shibahara Tomoyuki,Mase Masaji
BMC veterinary research
BACKGROUND:The type 2 highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) has spread throughout countries of southeast Asia, where it has caused severe economic losses. Even countries presently free of PRRSV are at high risk for infection and spread of this virus. Some of these countries, including Japan, have broad epidemics of the local type 2 PRRSV, creating chronic pathogenicity in the domestic pig population. The present study aimed to evaluate the protective efficacy of immunity by infection with a Japanese field isolate, EDRD1, against heterologous challenge with a Vietnamese HP-PRRSV field strain. To this end, four groups of PRRSV-negative crossbreed piglets were used for a challenge study. Groups 1 and 2 were inoculated with EDRD1 via the intranasal route. After 26 days, Groups 2 and 3 were inoculated with HP-PRRSV via the same route. Group 4 served as an uninfected control. Blood and oral fluid samples were taken every 3-4 days after HP-PRRSV challenge; on day 16 post-challenge, all pigs were euthanized, and examined pathologically. RESULTS:The nucleotide sequence analysis of nonstructural protein 2 gene of EDRD1 and comparison with Vietnamese HP-PRRSV showed that the 39 amino acid deletion sites of EDRD1 was nearly in the same region as the 29 amino acid deletion sites of HP-PRRSV. Immunity conferred by inoculation with EDRD1 dramatically reduced viral load in the sera and tissues besides viral shedding (Group 2) compared with those in pigs infected only with HP-PRRSV (Group 3). The clinical signs and rectal temperature were significantly reduced, and the average daily weight gain was significantly improved in the EDRD1-inoculated pigs (Group 2) compared with the Group 3 pigs. Notably, no viral RNA was detected in various organs of the Group 2 pigs 16 days post-infection with HP-PRRSV, except in one pig. Therefore, the immunity induced by EDRD1 and its genetically close field isolates may play a role in reducing viremia caused by HP-PRRSV. CONCLUSIONS:The results of the present study demonstrate that pigs are highly protected against heterologous Vietnamese HP-PRRSV challenge by immunity against a Japanese local strain, EDRD1.