Medicine and Health Sciences Commons^™

Host-Genetic-Based Outcome Of Co-Infection By Pcv2b And Prrsv In Pigs, Christian W. Eaton, Hiep Vu, Arabella L. Hodges, Seth P. Harris, Stephen D. Kachman, Daniel C. Ciobanu

School of Veterinary and Biomedical Sciences: Faculty Publications

Replication of porcine circovirus type 2 (PCV2), an important worldwide swine pathogen, has been demonstrated to be influenced by host genotype. Specifically, a missense DNA polymorphism (SYNGR2 p.Arg63Cys) within the SYNGR2 gene was demonstrated to contribute to variation in PCV2b viral load and subsequent immune response following infection. PCV2 is known to induce immunosuppression leading to an increase in susceptibility to subsequent infections with other viral pathogens such as porcine reproductive and respiratory syndrome virus (PRRSV). In order to assess the role of SYNGR2 p.Arg63Cys in co-infections, pigs homozygous for the favorable SYNGR2 p.63Cys (N = 30) …

Dynamics And Evolution Of Genotype 1 Porcine Reproductive And Respiratory Syndrome Virus Following Its Introduction Into A Herd Concurrently Infected With Genotypes 1 And 2, Thitima Tripipat, Kepalee Saeng-Chuto, Adthakorn Madapong, Christopher James Stott, Angkana Tantituvanont, Pavita Kaewprommal, Jittima Piriyapongsa, Dachrit Nilubol

The Thai Journal of Veterinary Medicine

This study was conducted to investigate the dynamics and evolution of porcine reproductive and respiratory syndrome virus (PRRSV) genotype 1 following its introduction into a herd concurrently infected with both PRRSV genotypes. The results demonstrated that 2 novel genotype 1 groups emerged, termed clusters II and III. The emergence of cluster II suggested that this cluster of genotype 1 virus potentially correlated with the outbreak. Then, cluster III was detected with the disappearance of cluster II. The evolution of the genotype 1 virus from cluster II to cluster III is likely to be associated with immune evasion induced by amino …

Quantification Of Semi-Truck Cab Decontamination, C. Grace Elijah, Cassandra K. Jones, Caitlin Evans, Haley K. Wecker, Charles R. Stark, Jianfa Bai, Elizabeth G. Poulsen-Porter, Allison K. Blomme, Jason C. Woodworth, Chad B. Paulk, Jordan T. Gebhardt

Kansas Agricultural Experiment Station Research Reports

Evidence suggests that the inside of vehicle cabs used for feed delivery may serve as a potential source for disease, yet there are no standardized protocols or scientific evidence for methods of their disinfection. Therefore, the objective of this project was to evaluate commercially available disinfectants and disinfection application methods against PEDV and PRRSV on various surfaces within semi-truck cabs. Three different surface types common in vehicle cabs (fabric, plastic, and rubber) were cut into 4 × 4 inch coupons and inoculated with either PEDV or PRRSV. Once inoculated, surfaces were placed in one of 3 semi-truck cabs and the …

Effect Of Oxidized Soybean Oil On The Immune Response To Porcine Reproductive And Respiratory Syndrome Modified Live Virus Vaccine In Nursery Pigs, Alongkot Boonsoongnern, Pichai Jirawattanapong, Win Surachetpong, Prapassorn Boonsoongnern, Pariwat Poolperm

The Thai Journal of Veterinary Medicine

This study evaluated the effect of oxidized soybean oil on the immune response to porcine reproductive and respiratory syndrome modified live vaccine (PRRS-MLV), and the oxidative stress status in nursery pigs.Seventycastrated weaned pigs, from a free PRRS virus infected herd, were divided into two groups: treatment group (n=63) and control group (n=7). The treatment pigs were vaccinated with the PRRS-MLV and then divided into three groups based on different diet types containing different soybean oils: (A) 5% fresh oil, (B) 5% heated oil for 43 h, and (C) 5% heated oil for 38 h. The control pigs were not vaccinated …

Detection Of Prrsv Circulation Using Oral Fluid Samples For Nursery Management In Endemically Prrsv-Infected Farms, Yonlayong Woonwong, Roongtham Kedkovid, Jirapat Arunorat, Chaitawat Sirisereewan, Teerawut Nedumpun, Korakrit Poonsuk, Yaowalak Panyasing, Pariwat Poolperm, Alongkot Boonsoongnern, Roongroje Thanawongnuwech

The Thai Journal of Veterinary Medicine

Porcine reproductive and respiratory syndrome (PRRS) has become a major swine disease worldwide. Relevant management strategies and diagnostic assays are of importance for PRRS virus (PRRSV) control. The objective of this study was to determine the use of oral fluids for PRRSV monitoring in endemically PRRSV-infected herds. PRRSV RNA and PRRSV-specific antibodies were monitored using oral fluid samples and serum samples in two conventional swine farms in Thailand (‘Farm A’, a one-site conventional system farm, and ‘Farm B’, a one-site P0-P1 segregation system farm) during farrowing to nursery periods. Both PRRSV RNA and PRRSV antibodies were detected from 3 to …

Expression Of Mir-29a And Ifn-Γ In Pigs Vaccinated With Modified Live Prrs Vaccine, Sunaree Nantakhruea, Nuntawan Aunjangwang, Orawan Limsivilai, Win Surachetpong

The Thai Journal of Veterinary Medicine

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the important RNA viruses that affects swine production worldwide. To better improve vaccine efficacy, understanding the molecular mechanisms and viral-host interaction is important to increase the knowledge of vaccine response. The objective of this study was to investigate the expression levels of miR-29a and IFN-γ in pigs that received modified live PRRSV vaccine. The study revealed that both miR-29a and IFN-γ transcript levels increased in vivo early, at day 3 post vaccination, whereas levels of miR-29a and IFN-γ mRNA were not different at day 7 post vaccination. Our study is …

Antiviral Activity Of Thymus Vulgaris And Nepeta Cataria Hydrosols Against Porcine Reproductive And Respiratory Syndrome Virus, Kraijak Kaewprom, Yu-Hsin Chen, Chuen-Fu Lin, Ming-Tang Chiou, Chao-Nan Lin

The Thai Journal of Veterinary Medicine

Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failure of sows and respiratory problems of nursery and growing pigs. Current management strategies mainly focus on preventing PRRS virus (PRRSV) infection through vaccination. However, the currently available modified live vaccines are not sufficient to eradicate the virus and do not provide complete immunity from infection. In search of agents that may be proven clinically effective against North American PRRSV (NA-PRRSV or US-PRRSV) infection, 42 natural herb hydrosols were purified and subjected to screening of their antiviral activity against two US-PRRSV strains in MARC-145 cells. Based on results obtained from …

Intranasal Immunization Of Pigs With Porcine Reproductive And Respiratory Syndrome Virus Like Particles Plus 2’, 3’ –Cgamp Vaccigrade™ Adjuvant Exacerbates Viremia After Virus Challenge, Alexandria Marie Van Noort

Electronic Theses and Dissertations

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically devastating virus that causes a persistent infection and spreads quickly. The virus causes abortions in sows and respiratory diseases in young piglets. Current PRRSV vaccines appear to provide limited cross-protection. A virus-like particles (VLPs) vaccine is made up of key PRRSV structural proteins, which could be cross protective among the different strains. We tested PRRSV VLPs as a vaccine candidate with a 2’, 3’-cGAMP VacciGradeTM adjuvant against pigs infected with PRRSV after intranasal immunization to evaluate the immune response and protective efficacy by the VLPs and adjuvant. The VLPs contain …

Cloning And Sequence Analysis Of Orf5 Gene Of Porcine Reproductive And Respiratory Syndrome Virus Of The Zj Strain, Baocheng Hao, Dengyuan Niu, Xiaoyong Xing, Haitao Xiang, Fengqin Wen, Xiaoping Fu, Jianping Liang, Yonghao Hu

The Thai Journal of Veterinary Medicine

A pair of the special primers was designed and synthesized according to the ORF5 sequence of porcine reproductive and respiratory syndrome virus (PRRSV), which was published in GenBank. In 2009 a pig farm in Zhejiang province suspected some diseased and dead pigs displaying symptoms of porcine reproductive and respiratory syndrome and provided 12 samples (spleen, kidney or lymph nodes) for laboratory diagnosis. The samples were positive of porcine reproductive and respiratory syndrome by PCR. The ORF5 gene was amplified by RT-PCR and cloned into pMD-18T vector. Nucleotide sequence was determined by sequencing and compared with other ORF5 genes in GenBank. …

Antiviral Activity Of Four Commercial Tilmicosin Preparations Against Porcine Reproductive And Respiratory Syndrome Virus (Prrsv): An In Vitro Study, Korakrit Poonsuk, Jirapat Arunorat, Yonlayong Woonwong, Panchan Sitthicharoenchai, Suphattra Jittimanee, Porjit Choojai, Roongroje Thanawongnuwech

The Thai Journal of Veterinary Medicine

The efficacy of four commercial tilmicosin preparations (A, B, C and D) against type 2 porcine reproductive and respiratory syndrome virus (PRRSV), 01NP1 (Thai isolate), infection in cultured pulmonary alveolar macrophages (PAMs) was conducted in vitro. Primary PAMs were collected from four 4-week-old PRRSV-free pigs. After separately plated onto four 24-well plates, PAMs were treated separately by 4 commercial tilmicosin preparations of 2 concentrations each (0.1 mg/ml and 0.01 mg/ml). The treated PAMs were inoculated with 0.05 MOI of 01NP1 strain and were stained with PRRSV specific antibody using immunoperoxidase monolayer assay (IPMA) to evaluate the quantity of PRRSV infected …

Molecular Characterization And Expression Analysis Of Mir-29a In Porcine Cells And Porcine Reproductive And Respiratory Syndrome Virus Infected Peripheral Blood Mononuclear Cells, Win Surachetpong, Sunaree Nantakhruea, Porntippa Lekcharoensuk

The Thai Journal of Veterinary Medicine

MicroRNAs are a group of small non-coding RNA that are involved in posttranscriptional regulation through target mRNA degradation or inhibition of protein synthesis. The objective of this present study was to characterize and determine expression level of porcine miR-29a in various tissues. Our results indicated that porcine miR-29a shared high conserved sequence with miR-29a from other organisms. Furthermore, in silico analysis suggested that the seed region of porcine miR-29a partially matched IFN-γ mRNA. RT-PCR analysis indicated that miR-29a expressed in various tissues including lymph node, spleen, peripheral blood mononuclear cells (PBMCs) and PK-15 cells. Expression profiling of miR-29a in PRRSV-infected …

Identification Of Amino Acid Residues Important For Anti-Ifn Activity Of Porcine Reproductive And Respiratory Syndrome Virus Non-Structural Protein 1, Lalit Beura, Sakthivel Subramaniam, Hiep Vu, Byungjoon Kwon, Asit K. Pattnaik, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

The non-structural protein 1 (nsp1) of porcine reproductive and respiratory syndrome virus is partly responsible for inhibition of type I interferon (IFN) response by the infected host. By performing alanine-scanning mutagenesis, we have identified amino acid residues in nsp1α and nsp1β~ (the proteolytic products of nsp1) that when substituted with alanine(s) exhibited significant relief of IFNsuppression. A mutant virus (16-SA, in which residues 16-20 of nsp1β were substituted with alanines) encoding mutant nsp1β recovered from infectious cDNA clone was shown to be attenuated for growth in vitro and induced significantly higher amount of type I IFN transcripts in infected macrophages. …

Amino Acid Residues In The Non-Structural Protein 1 Of Porcine Reproductive And Respiratory Syndrome Virus Involved In Down-Regulation Of Tnf-Cx Expression In Vitro And Attenuation In Vivo, Sakthivel Subramaniam, Lalit Beura, Byungjoon Kwon, Asit K. Pattnaik, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

Porcine reproductive and respiratory syndrome virus (PRRSV) suppresses tumor necrosis factor-alpha (TNF-α) production at both transcriptional and post-transcriptional levels by its non-structural proteins 1α and 1β (Nsp1α and Nsp1β). To identifY the amino acid residues responsible for this activity, we generated several alanine substitution mutants of Nsp1α and Nsp1β. Examination of the mutant proteins revealed that Nsp1α residues Gly90, Asn91 , Arg97, Argl 00 and Arg124 were necessary for TNF-α promoter suppression, whereas several amino acids spanning the entire Nsp1β ~ were found to be required for this activity. Two mutant viruses, with mutations at Nsp1α Gly90 or Nsp1β residues …

Role Of Porcine Reproductive And Respiratory Syndrome Virus Non-Structural Protein 1 In Viral Replication And Pathogenesis, Lalit Beura

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Porcine reproductive and respiratory syndrome virus (PRRSV) is an infectious agent of significant concern to the global swine industry. PRRSV infection of pigs is initiated by a long viremia after which the virus enters in an extended persistent phase of 3-4 months that ultimately is resolved by the immune system. The delayed and weak host adaptive response is responsible for the protracted convalescence period. An initial sub-optimal innate immune response is postulated to be the reason behind such meager adaptive immune response. A major focus of the studies undertaken in this dissertation was to identify the viral non-structural proteins (nsps) …

Molecular Structures Of Prrsv That Contribute To Prrsv Protective Immunity, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

One of the objectives of the proposal is to delineate the regions of the two glycoproteins (GP2 and GP4) of PRRSV that interact with CD163. Other objectives of the proposal are to generate antibodies to these small regions of the glycoproteins as well as to the full-length proteins for future studies to determine if any of these antibodies possess PRRSV neutralizing activity.

To carry out the studies in the proposed objectives, we generated a series of mutants of GP2 as well as GP4 proteins in which various regions were specifically removed by manipulating the plasmids encoding these proteins. We then …

The Increase Of Cd4+Cd25+ T Cells In The Peripheral Blood Of Pigs Persistently Infected With Porcine Reproductive And Respiratory Syndrome Virus, Jingwen Zeng, Danting He, Xin Guo, Xinna Ge, Hanchun Yang

Turkish Journal of Veterinary & Animal Sciences

Regulatory T lymphocytes (T-regs) may be involved in suppressing the host's immunity, which is important for viral clearance, and may therefore contribute to persistent viral infection. However, nothing is known regarding the situation of porcine T-regs during persistent infection with porcine reproductive and respiratory syndrome virus (PRRSV). This study, which used flow cytometry and real-time PCR, determined that the frequencies of CD4+CD25+ T-cells and porcine Forkhead/winged helix transcription factor (FoxP3) mRNA levels in peripheral blood significantly increased during PRRSV persistence, indicating that CD4+CD25+ T-regs might play an important role in PRRSV persistent infection. This study contributes to a better understanding …

The Glycoproteins Of Porcine Reproductive And Respiratory Syndrome Virus And Their Role In Infection And Immunity, Phani B. Das

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

The porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pathogen of swine and is known to cause abortion and infertility in pregnant sows and respiratory distress in piglets. PRRSV contains a major glycoprotein (GP5) and three minor glycoproteins (GP2a, GP3, and GP4) on the virion envelope, all of which are required for infectious virus production. To study their interactions amongst each other and with a cellular receptor for PRRSV, CD163, I cloned each of the viral glycoproteins and CD163 in various expression vectors. My studies have shown that while the GP2a, GP3, and GP4 are co-translationally glycosylated, …

All-Trans Retinoic Acid Regulates Intracellular Redox Status In Marc-145 Cells Infected With Porcine Reproductive And Respiratory Syndrome Virus, Yan Lin, De Wu

Turkish Journal of Veterinary & Animal Sciences

A relationship between oxidative stress, and apoptosis and/or death has been suggested for many years. The objective of this study was to determine the influence of all-trans retinoic acid (ATRA) on the cell survival rate and the antioxidant defense system. MARC-145 cells were treated with ATRA after infection with porcine reproductive and respiratory syndrome virus (PRRSV). The cell survival rate, total antioxidant capability (T-AOC), glutathione peroxidase (GSH-P_X) activity, superoxide dismutase (SOD) activity, lactate dehydrogenase (LDH) activity, and glutathione (GSH) content were determined in samples collected from cells incubated with ATRA for 24 or 48 h. The results show that PRRSV …

Gp3 Is A Structural Component Of The Prrsv Type Ii (Us) Virion, M. De Lima, I. H. Ansari, P. B. Das, B. J. Ku, F. J. Martinez-Lobo, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

Abstract Glycoprotein 3 (GP3) is a highly glycosylated PRRSV envelope protein which has been reported as being present in the virions of PRRSV type I, while missing in the type II PRRSV (US) virions. We herein present evidence that GP3 is indeed incorporated in the virus particles of a North American strain of PRRSV (FL12), at a density that is consistent with the minor structural role assigned to GP3 in members of the Arterivirus genus. Two 15aa peptides corresponding to two different immunodominant linear epitopes of GP3 derived from the North American strain of PRRSV (FL12) were used as antigen …

Mouse × Pig Chimeric Antibodies Expressed In Baculovirus Retain The Same Properties Of Their Parent Antibodies, Ana M. Jar, Fernando A. Osorio, Osvaldo J. López

School of Veterinary and Biomedical Sciences: Faculty Publications

The development of hybridoma and recombinant DNA technologies has made it possible to use antibodies against cancer, autoimmune disorders, and infectious diseases in humans. These advances in therapy, as well as immunoprophylaxis, could also make it possible to use these technologies in agricultural species of economic importance such as pigs. Porcine reproductive and respiratory syndrome virus (PRRSV) is an arterivirus causing very important economic losses to the industry. Passive transfer of antibodies obtained by biotechnology could be used in the future to complement or replace vaccination against this and other pig pathogens. To this end, we constructed and studied the …

Antibiotic-Mediated Inhibition Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Infection: A Novel Quinolone Function Which Potentiates The Antiviral Cytokine Response In Marc-145 Cells And Pig Macrophages, William A. Cafruny, Richard G. Duman, Raymond R. Rowland, Eric A. Nelson, Grace H. Wong

Veterinary and Biomedical Sciences Faculty Publications

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically significant agent for which there currently are no effective treatments. Development of antiviral agents for PRRSV as well as many other viruses has been limited by toxicity of known antiviral compounds. In contrast, antibiotics for non-virus microbial infections have been widely useful, in part because of their acceptable toxicity in animals. We report here the discovery that the quinolone containing compound Plasmocin™, as well as the quinolones nalidixic acid and ciprofl oxacin, have potent anti-PRRSV activity in vitro. PRRSV replication was inhibited by these antibiotics in both cultured MARC-145 cells …

The Detection Of Ifn-Γ Positive Cells In The Lymphoid Tissues Of Prrsv-Infected Pigs Using Different Strains, For Infection, Tuddow Thaiwong, Chanis Leetheraprasert, Chaiwat Winyoonunthakul, Anudep Rungsipipat, Supradit Wangnaitham, Roongroje Thanawongnuwech

The Thai Journal of Veterinary Medicine

Archived paraffin-embedded lymphoid tissue samples from seventy five experimental PRRSV-infected pigs were examined for PRRSV antigen and IFN-γ positive cells, using immunohistochemistry. Lymphoid tissues (5 pigs/group) including spleen, tracheobronchial lymph nodes and tonsil came from negative control pigs (group 1), low virulence strain of PRRSV (RespPRRS/ReproTM, modified-live virus vaccine) infected pigs (group 2) and high virulence strain of PRRSV (VR-2385) infected pigs(group 3). They were necropsied at 3, 7,10, 14 or 28 days post-inoculation (DPI). According to the study, microscopic lesions in the lymphoid tissues from pigs infected with the high virulence strain had more severe lesions, characterized by severe …

Antigen Distribution Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) In Thai Crossbred Pigs Using Immunohistochemistry, Porntip Laohasittikul, Nuttapon Boonarpha, Yongyut Pongprapachuen, Sawang Kesdangsakonwut, Supradit Wangnaitham, Roongroje Thanawongnuwech

The Thai Journal of Veterinary Medicine

Twenty, 3-week-old, crossbred pigs from a herd free from porcine reproductive and respiratory syndrome virus (PRRSV) were inoculated intranasally and intramusculary (1 ml each 102.5 TCID50/ml) with Thai PRRSV isolates of the US genotype (01NP1) (n=9) or the EU genotype (02SB3) (n=8). Three pigs were used as negative controls. Three infected and one control group pig were euthanatized 5, 9 or 15 days postinoculation (DPI) to detect and to compare the location and the amount of PRRSV-antigen present. Immunohistochemical evaluation of formalin-fixed tissues revealed PRRSV-antigen in the pulmonary macrophages of the lungs (6/9, 8/8 and 0/3 of the 01NP1-infected, 02SB3-infected …

Semiquantitative Analysis Of Il-10 Production In Bronchoalveolar Lavage Leukocytes Of Prrsv Infected Pigs, Permsak Thanawang, Chanida Yamkleebbua, Kritkong Shiyun, Roongroje Thanawongnuwech, Sawang Kesdangsakonwut, Supradit Wangnaitham, Sanipa Suradhat

The Thai Journal of Veterinary Medicine

Semi-quantitative analysis of the interleukin 10 (IL-10) in bronchoalveolar lavage cells (BALC), from pigs challenged with porcine reproductive and respiratory syndrome virus (PRRSV), was undertaken in this study. Twenty-one, three weeks old, crossbred pigs from a PRRSV-free commercial farm were separated into three groups, a control, non-infected group (3 pigs/group), and 2 groups (9 pigs/group), the first inoculated with a Thai-isolated European (O2SB3) strain and the second a Thai-isolated, North American (O1NP1) PRRSV strain. BALC were isolated from three infected pigs and one control on days 5, 9 and 15 post infection (DPI). The levels of IL-10 production in BALC …

Cytokine Profiles Following Mycoplasma Hyopneumoniae And Prrsv Co-Infection In Pigs, Roongroje Thanawongnuwech, Eileen L. Thacker

The Thai Journal of Veterinary Medicine

Bronchoalveolar cells collected by lung lavage 28 days post infection (DPI) of negative controls, Mycoplasma hyopneumoniae (M. hyo)-infected, Porcine Respiratory and Reproductive Syndrome virus (PRRSV)-infected and dual-infected pigs, were evaluated for various cytokine mRNA (IL1α, IL1β, IL6, IL8, IL10, IL12, and TNFα). Semiquantitative RT-PCR revealed that IL1β, IL6, IL10 and the IL12 mRNA expression was significantly increased in dual-infected pigs, compared with the negative controls. PRRSV-infected pigs had significantly increased levels of IL10 and IL12 mRNA. The increased IL10 gene expression that was observed in the dual infected pigs may play a role in the potentiation of PRRSV-induced pneumonia by …

The Role Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) In The Porcine Respiratory Disease Complex (Prdc), Roongroje Thanawongnuwech

The Thai Journal of Veterinary Medicine

Concurrent bacterial infections of the respiratory tract in porcine reproductive and respiratory syndrome virus (PRRSV)-infected pigs is a major problem for the swine industry and may involve various factors. This review manuscript discusses the role of pulmonary macrophages, which are affected by PRRSV. The macrophage's phagocytic and bactericidal ability is altered by the PRRSV infection as is pulmonary clearance. PRRSV also induces apoptosis and macrophages cell lysis leading to an increased susceptibility to secondary bacterial infection, and the syndrome called porcine respiratory disease complex (PRDC). Prevention and control strategies for these concurrent infections are briefly discussed.