Highly Pathogenic Porcine Reproductive And Respiratory Syndrome Research Articles

Recombinant Bivalent Live Vectored Vaccine Against Classical Swine Fever and HP-PRRS Revealed Adequate Heterogeneous Protection Against NADC30-Like Strain.

The recombinant bivalent live vectored vaccine rPRRSV-E2 has been proved to be a favorable genetic engineering vaccine against classical swine fever (CSF) and highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS). NADC30-like strains have recently emerged in China and caused severe disease, and it is necessary to evaluate the vaccine candidate for the currently circulating viruses. This study established a good challenge model to evaluate the candidate rPRRSV-E2 vaccine in preventing infection with a representative NADC30-like strain (ZJqz21). It was shown that the challenge control piglets displayed clinical signs typical of PRRSV, including a persistent fever, dyspnea, moderate interstitial pneumonia, lymph node congestion, and viremia. In contrast, the rPRRSV-E2 vaccination significantly alleviated the clinical signs, yielded a high level of antibodies, provided adequate protection against challenge with ZJqz21, and inhibited viral shedding and the viral load in target tissues. Our results demonstrated that the recombinant bivalent live vectored vaccine strain rPRRSV-E2 can provide efficient protection against the challenge of heterologous circulating NADC30-like strain and could be a promising vaccine candidate for the swine industry.

Molecular detection and characterization of highly pathogenic porcine reproductive and respiratory syndrome virus from a natural outbreak in wild pigs, Mizoram, India.

This study reports for the first time a natural outbreak of highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) caused by HP-PRRS virus (HP-PRRSV) in wild pigs characterized by sudden onset of depression, anorexia, respiratory distress, and high fever. The disease has caused severe haemorrhagic pneumonia, haemorrhagic lymphadenitis, enlarged spleen with areas of infarction, and petechial haemorrhages on the myocardium and on the surface of kidneys. HP-PRRSV was detected in representative tissue samples by reverse transcription-PCR, and the field strain was isolated in the MA104 cell line. The phylogenetic analyses based on the whole genome sequences and nucleotide sequences of open reading frame 5 (ORF5) gene showed close grouping with the subtype IV of lineage 8/8.7 of PRRSV II, which represents the HP-PRRSV strains that predominate in the pig population of China since 2010. The amino acid sequence analysis of the ORF5 gene revealed the replacement of leucine (L) at position 39 to isoleucine (I) in the primary neutralizing epitope. Among the four potential N glycosylation sites, the N34 was mutated and found to be restricted to only three N glycosylation sites. The present findings have indicated that HP-PRRSV can cause fatal outbreaks and may emerge as a major threat to the wild pig population.

The infectious cDNA clone of commercial HP-PRRS JXA1-R-attenuated vaccine can be a potential effective live vaccine vector.

Multiple commercial porcine reproductive and respiratory syndrome (PRRS) modified live vaccines are currently utilized in Chinese swine herds due to the limited cross-protection of vaccines and coexistence of different PRRS viruses. In this study, an infectious cDNA clone of the highly pathogenic PRRS (HP-PRRS) vaccine JXA1-R strain was generated. We successfully rescued the virus from direct in vitro DNA transfection of rJXA1-R clone, which has similar growth kinetics to the parental JXA1-R virus in Marc-145 cells. To further evaluate the potential use of the cloned rJXA1-R virus as a live vector for foreign gene expression, the enhanced green fluorescent protein (EGFP) was inserted between non-structural and structural genes. Our results showed that the dynamic expression of EGFP can be visualized by live cell imaging system during the infection in Marc-145 cells. The availability of our cloned JXA1-R viruses provides a crucial platform to study the fundamental biology of HP-PRRS virus vaccine and also serves as a potential effective vector for developing live vector vaccines against swine pathogens.

Construction and immunological evaluation of recombinant Newcastle disease virus vaccines expressing highly pathogenic porcine reproductive and respiratory syndrome virus GP3/GP5 proteins in pigs

Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) poses a significant threat to the pig industry, for which vaccination is considered to be an effective means of prevention and control. Here, we developed two recombinant Newcastle disease virus (NDV) LaSota-vectored PRRS candidate vaccines, rLaSota-GP5 and rLaSota-GP3-GP5, using reverse genetic techniques. The two recombinant viruses exhibited a high degree of genetic stability after 10 successive generations in chicken embryos. There was no significant difference in pathogenicity compared with the rLaSota parent strain in poultry, mice and pigs. The recombinant viruses could not be detected in the feeding environment of immunized pigs, but could be detected in the organs and tissues of pigs for no more than 10 days after immunization. Importantly, in contrast to rLaSota-GP5, rLaSota-GP3-GP5 elicited both significant humoral and cellular immune responses in pigs. In particular, the neutralizing antibody titer in the rLaSota-GP3-GP5 group was 1.51 times significantly higher than that of the commercial vaccine group at 42 days post-immunization. At the same time, there was significant difference in the level of IFN-γ between the rLaSota-GP3-GP5 group and the commercial vaccine group. Furthermore, the viral load in the organs and tissues of rLaSota-GP3-GP5-immunized pigs was substantially lower than that of unimmunized pigs after being challenged with HP-PRRS virus GD strain. These results suggest that rLaSota-GP3-GP5 is a safe and promising candidate vaccine, and there is potential for further development of a recombinant virus vaccine for PRRS using NDV.

The emergence of a highly pathogenic porcine reproductive and respiratory syndrome virus with additional 120aa deletion in Nsp2 region in Jiangxi, China.

Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS), which emerged in China in 2006, was characterized by high fever, high morbidity and high mortality. The causative agent of the disease was a highly pathogenic variant of porcine reproductive and respiratory syndrome virus (also called HP-PRRSV), which has a discontinuous deletion of 1+29 amino acids (aa) in the Nsp2 coding region, compared to classical PRRSV. In 2014, fattened pigs on a pig farm in Jiangxi Province suffered from clinical symptoms of high fever, dyspnoea and death. A PRRSV, termed JX2014T2, was isolated from samples of the dead pigs. Genomic analysis of the isolated PRRSV indicated that the genome of the virus was 14,960bp in length and belonged to the North American genotype. In the Nsp2-coding region, there was a discontinuous deletion of 1+29 aa, similar to HP-PRRSV; however, an additional continuous deletion of 120 amino acids between aa 628 and 747 was found. Further analysis of the pathogenicity of PRRSV JX2014T2 was performed in piglets, and the results indicated that all infected piglets suffered from typical clinical symptoms of PRRS, such as high fever, cough, mental depression, anorexia, dyspnoea and palpebral swelling and died within 15days postinfection (dpi). This demonstrated that the newly isolated PRRSV JX2014T2 strain containing an additional deletion of 120 aa is highly pathogenic to piglets, suggesting that a highly pathogenic variant with new genetic features is circulating in China.

Efficacy of Type 2 porcine reproductive and respiratory syndrome virus (PRRSV) vaccine against the 2010 isolate of Vietnamese highly pathogenic PRRSV challenge in pigs.

The efficacy of a commercial attenuated live type 2 porcine reproductive and respiratory syndrome (PRRS) vaccine was tested under experimental infection with a highly virulent Vietnamese virus isolated from a diseased pig affected with highly pathogenic PRRS (HP-PRRS) using specific pathogen-free (SPF) pigs. Twenty-five 4-week-old SPF pigs were divided into three groups as follows: pigs vaccinated with a single dose of the vaccine (Group 1, n=10), unvaccinated pigs (Group 2, n=10) and unvaccinated and non-infectious control pigs (Group 3, n=5). Four weeks later, Groups 1 and 2 were challenged with a 1 ml inoculum containing 1 × 105.5 50% tissue culture infectious dose (TCID50)/ml of a Vietnamese HP-PRRS virus isolated in 2010 via the intranasal route. Animals were monitored during the subsequent two-week period post-challenge and necropsied for virological and pathological assays. Results showed a significant reduction in viral replication and shedding in vaccinated pigs compared to unvaccinated pigs. The non-vaccinated pigs showed severe pyrogenic and respiratory illness with marked systematic lesions including interstitial pneumonia and thymic atrophy. In contrast, vaccinated pigs recovered quickly from fever with only mild pathological manifestations. Therefore, although viral shedding was still noted, immunization with the live PRRS vaccine did indeed reduce viral replication and disease severity, suggesting its utility in minimizing outbreaks of HP-PRRS.

Comparative Respiratory Pathogenicity and Dynamic Tissue Distribution of Chinese Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus and its Attenuated Strain in Piglets

The outbreak of highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) in 2006 devastated the Chinese swine industry. HP-PRRS virus is still the predominant strain in mainland China, rather than the classical PRRSV strain, and the attenuated live vaccine remains the preferred choice for protecting piglets against HP-PRRSV infection. To fully evaluate the safety of strain GDr180, the 180th attenuated virus of the HP-PRRSV strain GD, we used clinicopathological, microscopical, ultrastructural, serological and molecular biological methods to assess the different clinical manifestations and respiratory characteristics of piglets inoculated with HP-PRRSV strain GD or strain GDr180. The 5-week-old piglets inoculated with strain GD displayed marked clinical signs, including fever, anorexia, dyspnoea and tachypnoea. Significant interstitial pneumonia was present, characterized by thickened alveolar septa infiltrated with mononuclear cells and cell debris. However, the piglets inoculated with strain GDr180 and the negative control piglets showed neither clinical signs nor microscopical or ultrastructural lesions. Ultrastructural observation of the piglets' tracheas and examination of the dynamic tissue distributions of PRRSV strain GD and attenuated strain GDr180, by immunohistochemistry and fluorescence quantitative reverse transcription-polymerase chain reaction, confirmed significant differences in their pathogenicity and distribution in the respiratory systems of piglets. The differences in pathogenicity are attributable to the different severity of the pathological changes in the pigs inoculated with the two strains. Thus, the HP-PRRSV GDr180 strain is practically harmless to the respiratory systems of piglets and may be a safe candidate for inducing immunity against HP-PRRS.

Emergence of highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) in medium-scale swine farms in southeastern Cambodia

Since 2006, reports from China and Viet Nam have alerted of an emergent highly pathogenic variant of porcine reproductive and respiratory syndrome virus (HP-PRRSV) in that region. The frequent occurrence of outbreaks in these countries puts Cambodian pig farms at high risk of infection, but no study had been conducted to investigate the presence of HP-PRRS in Cambodian farms. We investigated the presence of HP-PRRS in medium-scale (semi-commercial) swine farms in the Cambodian southeastern region. Specifically, one province bordering Viet Nam (Takeo) was selected due to the concentration of most semi-commercial farms in that province. A cross-sectional study was carried out, between July and September 2010 to assess whether the prevalence of infection in these farms was indicative of recent spread of PPRSV and to identify risk factors for infection. The number of farms to be sampled was established using methods for Lot Quality Assurance Surveys (LQAS), in order to achieve a pre-established ability to discriminate between two different prevalence settings.The target population comprised all semi-commercial farms in Takeo province from which a random sample of 35 farms was selected. Selected farms were visited and questionnaires administered to gather information on farm characteristics and husbandry practices. Blood samples from individual pigs were collected in each of the study farms and tested for PRRSV, along with a number of other swine respiratory pathogens in order to investigate potential interactions.Our results showed that the virus was already present in Takeo semi-commercial pig population (LQAS herd prevalence ≥85%) at the time of sampling. The presence of sows in the farm and farm density were significantly associated (P<0.05) with the introduction and the presence of PRRS – but this was an unadjusted association as small sample size precluded multivariate analysis. Spatiotemporal description of the supposed pattern of infection revealed that the 1st farms infected were closely located to major national and provincial roads, connecting the Cambodian capital Phnom Penh to Viet Nam.

Simultaneous detection and differentiation of highly virulent and classical Chinese-type isolation of PRRSV by real-time RT-PCR.

Porcine reproductive and respiratory syndrome (PRRS) is a leading disease in pig industry worldwide and can result in serious economic losses each year. The PRRS epidemic situation in China has been very complicated since the unprecedented large-scale highly pathogenic PRRS (HP-PRRS) outbreaks in 2006. And now the HP-PRRS virus (HP-PRRSV) and classical North American type PRRSV strains have coexisted in China. Rapid differential detection of the two strains of PRRSV is very important for effective PRRS control. The real-time RT-PCR for simultaneous detection and differentiation of HP-PRRSV and PRRSV by using both SYBR Green and TaqMan probes was developed and validated. Both assays can be used for rapid detection and strain-specific identification of HP-PRRSV and PRRSV. However, the TaqMan probe method had the highest detection rate whereas the conventional RT-PCR was the lowest. The real-time RT-PCR developed based on SYBR Green and TaqMan probe could be used for simultaneous detection and differentiation of HP-PRRSV and PRRSV in China, which will benefit much the PRRS control and research.

New genomic characteristics of highly pathogenic porcine reproductive and respiratory syndrome viruses do not lead to significant changes in pathogenicity

Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) initially emerged in China and currently prevails in other Asian countries as well, resulting in immense economic losses. HP-PRRS virus (HP-PRRSV) has undergone rapid evolution since its first recognition in 2006. To analyze the genomic and pathogenic characteristics of 2010 HP-PRRSV, we tested 919 clinical samples collected from China, Laos and Vietnam, sequenced 29 complete genomes of HP-PRRSV isolates, and determined the pathogenicity of seven HP-PRRS viruses isolated from 2006 to 2010. HP-PRRSV was detected from 45.2% (415/919) samples, while only 0.1% (1/919) was classical PRRSV, indicating that HP-PRRSV isolates with a unique discontinuous deletion of 30 amino acids (aa) in non-structural protein 2 (Nsp2) are still the predominant viruses. 2010 HP-PRRSV together with 2009 HP-PRRSV isolates form a new evolutionary branch based on phylogenetic analyses. The numbers of potential N-glycosylation sites are variable in major glycoprotein GP5 but are conserved in minor glycoproteins GP2, GP3 and GP4. Pathogenicity studies showed that HP-PRRS viruses isolated from 2006 to 2010 maintain similar level of high pathogenicity, which caused high fever (>41°C for at least four days), 100% morbidity, and 40–100% mortality in 4–10 weeks old pigs. Real time monitoring information from this study could help to understand the genetic and pathogenic evolution of HP-PRRSV and assist in the control of HP-PRRS in Asia.

One world--one health: the threat of emerging swine diseases. An Asian perspective.

SummaryOwing to the expanding globalization, the trans‐boundary spread of an epizootic can easily result from uncontrolled animal movements and human traffic. Foot and mouth disease (FMD) is a major trans‐boundary disease in most Asian countries. Its sporadic re‐emergence suggests that collaborative FMD control strategies should be uniformly implemented in endemic countries to ensure the overall national herd vaccination coverage, biocontainment when outbreaks occur, and strict biosecurity control of animal movement between countries. Sustained commitments from governments, cooperative diplomatic relationships, and public awareness campaigns are critical to FMD control, to ensure collaboration among veterinarians, traders and farmers throughout Southeast Asia (SEA). Recently, highly pathogenic porcine reproductive and respiratory syndrome (HP‐PRRS) and porcine epidemic diarrhoea (PED) spread from China to Southeast Asian countries, causing major economic losses. Foot and mouth disease, HP‐PRRS, and PED currently remain endemic and may continue to sporadically re‐emerge, owing to inadequate public health management and/or biosecurity failures. Therefore, the risk factors must be identified to better understand the epidemiology of these diseases in an effort to develop effective control measures. International coordination through the establishment of a collaborative network supported by the World Organization for Animal Health (OIE) and the Food and Agriculture Organization (FAO) should be implemented to prevent trans‐boundary transmission among countries. This review discusses trans‐boundary swine diseases of particular importance to SEA, including FMD, HP‐PRRS and PED, with a primary focus on major factors contributing to the spread of these diseases and important control measures, reflecting the impact of globalization on disease control and surveillance.

Emergence and pathogenicity of highly pathogenic Porcine reproductive and respiratory syndrome virus in Vientiane, Lao People’s Democratic Republic

Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) characterized by high fever, high morbidity, and high mortality in pigs of all ages emerged in China in 2006 and spread rapidly throughout Southeast Asia. In July 2010, a highly contagious swine disease with clinical signs similar to those of HP-PRRS was observed in the Lao People's Democratic Republic (Lao PDR). A field investigation covering 8 pig farms and 1 slaughterhouse in 7 different districts in the capital city of Vientiane was conducted to identify the disease. Total mortality rates ranged from 6.02% in boars to 91.28% in piglets (mean 54.15%) across the farms investigated. Emergence of the HP-PRRS virus (HP-PRRSV) in Lao PDR was confirmed using real-time reverse transcription polymerase chain reaction as well as virus isolation and identification. An animal inoculation study was performed to characterize the HP-PRRSV responsible for this outbreak. Isolate Laos 1.13 was inoculated into 70-day-old specific pathogen-free pigs to study pathogenicity. Clinical signs of high fever, rubefaction, respiratory distress, nervous symptoms, and diarrhea were observed in inoculated pigs, as well as pathological hemorrhagic lesions consolidated in the lungs. Morbidity and mortality were 100% and 60%, respectively, in inoculated pigs. HP-PRRSV was re-isolated from the inoculated pigs. Results suggested that the newly emerged HP-PRRSV was responsible for recent outbreaks of the swine disease in Lao PDR. The current report highlights the importance of continuous surveillance in neighboring countries to prevent introduction of PRRS to new regions.

Mutations in the genome of the highly pathogenic porcine reproductive and respiratory syndrome virus potentially related to attenuation

A live-attenuated highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) virus (HP-PRRSV) TJM vaccine strain was derived from the HP-PRRSV TJ strain by passage 92 times in the African green monkey kidney epithelial cell line Marc-145. We found that the virulence of the TJ strain to piglets was decreased greatly from passage 19. To identify mutations associated with attenuation of the TJM vaccine strain, we determined the nucleotide changes that arose during Marc-145 passage of the HP-PRRSV TJ virus. The TJM strain contains a 360 nucleotide (120 amino acids) deletion and a 118 nucleotide mutation that resulted in 48 amino acid changes. Analysis of the complete nucleotide sequences of intermediate passage-level viruses F19, F46 and F78 showed that 31 (64.6%) of the 48 amino acid mutations occurred in F19, 7 (14.6%) occurred in F46, 7 (14.6%) occurred in F78 and 3 (6.3%) occurred in F92. The 120 amino acid deletion occurred from F19 to TJM. Therefore, we hypothesized that the 31 amino acid mutations distributed in nsp1β, nsp2–nsp5, nsp7, nsp9, nsp10, GP4 and GP5 and the continuous 120 amino acid deletion in the nsp2 region from F19 provide a strong potential molecular basis for the observed attenuated phenotype.

The epidemic status and genetic diversity of 14 highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) isolates from China in 2009

A high-mortality swine disease, the highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS), reappeared in some regions of China in 2009. To explore the possible mechanisms underlying the emergence of HP-PRRSV and more fully understand the extent of the genetic diversity of this virus in China, the complete genome of 14 isolates from 10 provinces in China from 2009 were analyzed. Full-length genome sequencing analysis showed that the 14 isolates were closely related to HP-PRRSV, with 98.0–98.9% nucleotide similarity, although 2 of the 14 strains exhibited a new, discontinuous 29-amino acid deletion in the Nsp2 gene. Furthermore, amino acid analysis of the GP5 protein indicated that the 14 isolates had a concurrent mutation in a decoy epitope and different mutations in glycosylation sites. Additionally, the antigenic drift in GP3 and a 1-nucleotide deletion in both the 5′-UTR and 3′-UTR, which are found in almost all highly pathogenic Chinese PRRSV isolates, were examined in all 14 isolates. The phylogenetic analysis showed that the 14 strains belonged to the North American genotype and were clustered in a subgroup with other HP-PRRSV isolates that have been found in China since 2006. However, compared with other Chinese HP-PRRSV isolates collected in 2006–2008, the phylogenetic tree showed that the 14 isolates had a closer relationship with each other. These results indicated that HP-PRRSV remained an extensive pandemic, affecting swine farms in China in 2009 and revealed new genetic diversity.