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Sample records for virulent porcine reproductive

  1. Passive transfer of virus-specific antibodies confers protection against reproductive failure induced by a virulent strain of porcine reproductive and respiratory syndrome virus and establishes sterilizing immunity.

    Science.gov (United States)

    Osorio, F A; Galeota, J A; Nelson, E; Brodersen, B; Doster, A; Wills, R; Zuckermann, F; Laegreid, W W

    2002-10-10

    Immune mechanisms mediating protective immunity against porcine reproductive and respiratory syndrome virus (PRRSV) are not well understood. The PRRSV-specific humoral immune response has been dismissed as being ineffective and perhaps deleterious for the host. The function of PRRSV antibodies in protective immunity against infection with a highly abortifacient strain of this virus was examined by passive transfer experiments in pregnant swine. All of a group of pregnant gilts (n = 6) that received PRRSV immunoglobulin (Ig) from PRRSV-convalescent, hyperimmune animals were fully protected from reproductive failure as judged by 95% viability of offspring at weaning (15 days of age). On the other hand, the totality of animals in a matched control group (n = 6) receiving anti-pseudorabies virus (PRV) Ig exhibited marked reproductive failure with 4% survival at weaning. Besides protecting the pregnant females from clinical reproductive disease, the passive transfer of PRRSV Ig prevented the challenge virus from infecting the dams and precluded its vertical transmission, as evidenced by the complete absence of infectious PRRSV from the tissues of the dams and lack of infection in their offspring. In summary, these results indicate that PRRSV-Igs are capable of conferring protective immunity against PRRSV and furthermore that these Igs can provide sterilizing immunity in vivo.

  2. Comparative virulence of reproductive diseases caused by type 1 (European-like) and type 2 (North American-like) porcine reproductive and respiratory syndrome virus in experimentally infected pregnant gilts.

    Science.gov (United States)

    Han, K; Seo, H W; Park, C; Kang, I; Youn, S-K; Lee, S Y; Kim, S-H; Chae, C

    2014-01-01

    The aim of this study was to compare the virulence of type 1 and type 2 porcine reproductive and respiratory syndrome virus (PRRSV) as assessed by the level of viral replication, viral distribution and apoptosis in stillborn fetuses and live-born piglets from infected pregnant gilts. Type 1 or type 2 PRRSV was given intranasally to pregnant gilts at 3 weeks before the expected date of parturition. Regardless of virus genotype, PRRSV-infected gilts farrowed between 102 and 109 days of gestation, while control uninfected gilts carried the pregnancy to term and farrowed at 114-115 days of gestation. There were no significant differences in the mean number of virus-infected cells per unit area of tissue when type 1 and type 2 virus infections were compared between stillborn fetuses and live-born piglets. Stillborn fetuses from the type 1 PRRSV-infected pregnant gilts had a significantly higher mean number of apoptotic cells per unit area of thymus (P = 0.013) than those from type 2 PRRSV-infected pregnant gilts. Significant differences in virulence were not observed between types 1 and 2 PRRSV in terms of female reproductive failure, although thymic apoptosis differed in stillborn fetuses from type 1 and type 2 PRRSV-infected pregnant gilts. Copyright © 2014. Published by Elsevier Ltd.

  3. An interferon inducing porcine reproductive and respiratory syndrome virus vaccine candidate elicits protection against challenge with the heterologous virulent type 2 strain VR-2385 in pigs.

    Science.gov (United States)

    Fontanella, Eve; Ma, Zexu; Zhang, Yanjin; de Castro, Alessandra M M G; Shen, Huigang; Halbur, Patrick G; Opriessnig, Tanja

    2017-01-03

    Achieving consistent protection by vaccinating pigs against porcine reproductive and respiratory syndrome virus (PRRSV) remains difficult. Recently, an interferon-inducing PRRSV vaccine candidate strain A2MC2 was demonstrated to be attenuated and induced neutralizing antibodies. The objective of this study was to determine the efficacy of passage 90 of A2MC2 (A2P90) to protect pigs against challenge with moderately virulent PRRSV strain VR-2385 (92.3% nucleic acid identity with A2MC2) and highly virulent atypical PRRSV MN184 (84.5% nucleic acid identity with A2MC2). Forty 3-week old pigs were randomly assigned to five groups including a NEG-CONTROL group (non-vaccinated, non-challenged), VAC-VR2385 (vaccinated, challenged with strain VR-2385), VR2385 (challenged with strain VR-2385), VAC-MN184 (vaccinated, challenged with strain MN184) and a MN184 group (challenged with MN184 virus). Vaccination was done at 3weeks of age followed by challenge at 8weeks of age. No viremia was detectable in any of the vaccinated pigs; however, by the time of challenge, 15/16 vaccinated pigs had seroconverted based on ELISA and had neutralizing antibodies against a homologous strain with titers ranging from 8 to 128. Infection with VR-2385 resulted in mild-to-moderate clinical disease and lesions. For VR-2385 infected pigs, vaccination significantly lowered PRRSV viremia and nasal shedding by 9days post challenge (dpc), significantly reduced macroscopic lung lesions, and significantly increased the average daily weight gain compared to the non-vaccinated pigs. Infection with MN184 resulted in moderate-to-severe clinical disease and lesions regardless of vaccination status; however, vaccinated pigs had significantly less nasal shedding by dpc 5 compared to non-vaccinated pigs. Under the study conditions, the A2P90 vaccine strain was attenuated without detectable shedding, improved weight gain, and offered protection to the pigs challenged with VR-2385 by reduction of virus load and

  4. Attenuation of virulence of Lawsonia intracellularis after in vitro passages and its effects on the experimental reproduction of porcine proliferative enteropathy.

    Science.gov (United States)

    Vannucci, Fabio A; Beckler, Dana; Pusterla, Nicola; Mapes, Samantha M; Gebhart, Connie J

    2013-02-22

    Non-pathogenic Lawsonia intracellularis variants have been obtained through multiple passages in cell culture but there is no information regarding the number of passages necessary to attenuate a pathogenic isolate. The present study evaluated the susceptibility of pigs to L. intracellularis after 10, 20 and 40 passages in vitro. Three groups (six animals/group) were inoculated with pure culture of L. intracellularis on passage 10, 20 or 40 and one group with placebo. The animals were monitored for clinical signs, fecal shedding and serological IgG response during 28 days post-inoculation. Gross and histologic lesions and the level of infection based on the amount of L. intracellularis-specific antigen in the intestinal mucosa identified by immunohistochemistry were evaluated in two animals from each group on days 14, 21 and 28. Animals inoculated with passages 10 and 20 demonstrated proliferative lesions typical of porcine proliferative enteropathy associated with the presence of Lawsonia-specific antigen in the intestinal mucosa. Passage 40-inoculated pigs did not show proliferative lesions or presence of Lawsonia antigen at any time point throughout the study. Similar patterns of the fecal shedding were observed in passage 10 and 20-infected pigs but those infected with passage 40 shed for a short period. Serological IgG responses in passage 10 and 20-inoculated pigs were detected from day 14 post-infection but not at all in passage 40-inoculated animals. These results demonstrate attenuation of the virulence properties of L. intracellularis between 20 and 40 cell passages in vitro. This information will be valuable for design of future experimental models and for studying the mechanisms involved in the attenuation of L. intracellularis virulence. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)

    DEFF Research Database (Denmark)

    Kvisgaard, Lise Kirstine

    This PhD thesis presents the diversity of Porcine Reproductive and Respiratory Syndrome viruses (PRRSV) circulating in the Danish pig population. PRRS is a disease in pigs caused by the PRRS virus resulting in reproductive failures in sows and gilts and respiratory diseases in pigs . Due to genetic...

  6. Evidence of porcine reproductive and respiratory syndrome virus ...

    African Journals Online (AJOL)

    Evidence of porcine reproductive and respiratory syndrome virus antibodies in commercial pig husbandry in Nigeria: a preliminary study. ... main economic resources of developing countries. In Nigeria, pig farming contributes significantly to the livestock sub-sector and accounts for about 30% of total pig production in Africa.

  7. Challenges for porcine reproductive and respiratory syndrome virus (PRRSV) vaccinology

    NARCIS (Netherlands)

    Kimman, T.G.; Cornelissen, A.H.M.; Moormann, R.J.M.; Rebel, J.M.J.; Stockhofe, N.

    2009-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be a threat for the pig industry. Vaccines have been developed, but these failed to provide sustainable disease control, in particular against genetically unrelated strains. Here we give an overview of current knowledge and

  8. Spatial epidemiology of porcine reproductive and respiratory syndrome in Thailand.

    Science.gov (United States)

    Thanapongtharm, Weerapong; Linard, Catherine; Pamaranon, Nutavadee; Kawkalong, Sarayuth; Noimoh, Tanom; Chanachai, Karoon; Parakgamawongsa, Tippawon; Gilbert, Marius

    2014-08-05

    Porcine reproductive and respiratory syndrome (PRRS) has become a worldwide endemic disease of pigs. In 2006, an atypical and more virulent PRRS (HP-PRRS) emerged in China and spread to many countries, including Thailand. This study aimed to provide a first description of the spatio-temporal pattern of PRRS in Thailand and to quantify the statistical relationship between the presence of PRRS at the sub-district level and a set of risk factors. This should provide a basis for improving disease surveillance and control of PRRS in Thailand. Spatial scan statistics were used to detect clusters of outbreaks and allowed the identification of six spatial clusters covering 15 provinces of Thailand. Two modeling approaches were used to relate the presence or absence of PRRS outbreaks at the sub-district level to demographic characteristics of pig farming and other epidemiological spatial variables: autologistic multiple regressions and boosted regression trees (BRT). The variables showing a statistically significant association with PRRS presence in the autologistic multiple regression model were the sub-district human population and number of farms with breeding sows. The predictive power of the model, as measured by the area under the curve (AUC) of the receiver operating characteristics (ROC) plots was moderate. BRT models had higher goodness of fit the metrics and identified the sub-district human population and density of farms with breeding sows as important predictor variables. The results indicated that farms with breeding sows may be an important group for targeted surveillance and control. However, these findings obtained at the sub-district level should be complemented by farm-level epidemiological investigations in order to obtain a more comprehensive view of the factors affecting PRRS presence. In this study, the outbreaks of PRRS could not be differentiated from the potential novel HP-PPRS form, which was recently discovered in the country.

  9. Virulence-associated gene pattern of porcine and human Yersinia enterocolitica biotype 4 isolates.

    Science.gov (United States)

    Schneeberger, M; Brodard, I; Overesch, G

    2015-04-02

    Yersinia enterocolitica 4/O:3 is the most important human pathogenic bioserotype in Europe and the predominant pathogenic bioserotype in slaughter pigs. Although many studies on the virulence of Y. enterocolitica strains have showed a broad spectrum of detectable factors in pigs and humans, an analysis based on a strict comparative approach and serving to verify the virulence capability of porcine Y. enterocolitica as a source for human yersiniosis is lacking. Therefore, in the present study, strains of biotype (BT) 4 isolated from Swiss slaughter pig tonsils and feces and isolates from human clinical cases were compared in terms of their spectrum of virulence-associated genes (yadA, virF, ail, inv, rovA, ymoA, ystA, ystB and myfA). An analysis of the associated antimicrobial susceptibility pattern completed the characterization. All analyzed BT 4 strains showed a nearly similar pattern, comprising the known fundamental virulence-associated genes yadA, virF, ail, inv, rovA, ymoA, ystA and myfA. Only ystB was not detectable among all analyzed isolates. Importantly, neither the source of the isolates (porcine tonsils and feces, humans) nor the serotype (ST) had any influence on the gene pattern. From these findings, it can be concluded that the presence of the full complement of virulence genes necessary for human infection is common among porcine BT 4 strains. Swiss porcine BT 4 strains not only showed antimicrobial susceptibility to chloramphenicol, cefotaxime, ceftazidime, ciprofloxacin, colistin, florfenicol, gentamicin, kanamycin, nalidixic acid, sulfamethoxazole, streptomycin, tetracycline and trimethoprim but also showed 100% antibiotic resistance to ampicillin. The human BT 4 strains revealed comparable results. However, in addition to 100% antibiotic resistance to ampicillin, 2 strains were resistant to chloramphenicol and nalidixic acid. Additionally, 1 of these strains was resistant to sulfamethoxazole. The results demonstrated that Y. enterocolitica BT 4

  10. Recurrent reproductive failure associated with porcine reproductive and respiratory syndrome in a swine herd.

    Science.gov (United States)

    Dee, S A; Joo, H S

    1994-10-01

    An investigation of a case of recurrent reproductive failure in a swine herd was conducted. The subject farm (A) and a neighboring farm (B) had experienced an acute outbreak of porcine reproductive and respiratory syndrome in 1990. During 1992, farm A experienced recurrent episodes of conception failure, stillbirths, preweaning mortality, and sow mortality. Similar clinical findings were not evident on farm B. Evaluation of production records indicated that the problem was occurring in first-parity litters. It was learned that replacement gilts for farm A were being purchased from an outside source, whereas farm B raised its own replacement gilts. Indirect fluorescent antibody testing of gilts from both farms indicated a high prevalence of seronegative gilts on farm A vs a high prevalence of seropositive gilts on farm B. Although there is no correlation between indirect fluorescent antibody titers and immunity, it would appear that exposure to porcine reproductive and respiratory syndrome virus prior to breeding is essential to prevent reproductive disease.

  11. Effector mechanisms of humoral immunity to porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Rahe, Michael C; Murtaugh, Michael P

    2017-04-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) continues to afflict swine nearly 30 years after it was first discovered as the causative agent of "mystery swine disease". Immunological tools of vaccination and exposure to virulent viruses have not succeeded in achieving control and prevention of PRRSV. Humoral immunity, mediated by antibodies, is a hallmark of anti-viral immunity, but little is known about the effector mechanisms of humoral immunity against PRRSV. It is essential to understand the immunological significance of antibody functions, including recently described broadly neutralizing antibodies and potential non-neutralizing activities, in the immune response to PRRSV. Here, we review recent research from PRRSV and other host-pathogen interactions to inform novel routes of exploration into PRRSV humoral immunity which may be important for identifying the immunological correlates of protection against PRRSV infection. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Birth weight, intrauterine growth retardation and fetal susceptibility to porcine reproductive and respiratory syndrome virus

    Science.gov (United States)

    The severity of porcine reproductive and respiratory syndrome was compared in pregnant gilts originating from high and low birth weight litters. One-hundred and eleven pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus on gestation day 85 (±1) were necrop...

  13. Use of polarized light microscopy in porcine reproductive technologies.

    Science.gov (United States)

    Caamaño, J N; Maside, C; Gil, M A; Muñoz, M; Cuello, C; Díez, C; Sánchez-Osorio, J R; Martín, D; Gomis, J; Vazquez, J M; Roca, J; Carrocera, S; Martinez, E A; Gómez, E

    2011-09-01

    The meiotic spindle in the oocyte is composed of microtubules and plays an important role during chromosome alignment and separation at meiosis. Polarized light microscopy (PLM) could be useful for a non-invasive evaluation of the meiotic spindle and may allow removal of nuclear structures without fluorochrome staining and ultraviolet exposure. In this study, PLM was used to assess its potential application in porcine reproductive technologies. The objectives of the present study were to assess the efficiency of PLM to detect microtubule-polymerized protein in in vitro-matured porcine oocytes; to examine its effects on the oocyte developmental competence; to select oocytes based on the presence of the meiotic spindle detected by PLM; and to assess the efficiency oocyte enucleation assisted with PLM. In the first experiment, the presence of microtubule-polymerized protein was assessed and confirmed in oocytes (n = 117) by immunostaining and chromatin detection. In the second experiment, oocytes (n = 160) were exposed or not (controls) to PLM for 10 minutes, and then parthenogenetically activated and cultured in vitro. In the third experiment, development competence of oocytes with a positive or negative signal to PLM was analyzed after in vitro fertilization. Finally, oocytes (n = 54) were enucleated using PLM as a tool to remove the meiotic spindle. A positive PLM signal was detected in 98.2 % of the oocytes, which strongly correlated (r = 1; p PLM did not differ significantly from controls on cleavage, total blastocyst, expanded blastocyst rates and total cell numbers. The percentage of oocytes at the MII stage and blastocyst formation rate in the negative PLM group significantly differed from control and PLM positive groups. Overall efficiency of spindle removal using the PLM-Oosight system was 92.6%. These results suggest that polarized light microscopy is an efficient system to detect microtubule-polymerized protein in in vitro-matured porcine oocytes and does

  14. Automated 5 ' nuclease assay for detection of virulence factors in porcine Escherichia coli

    DEFF Research Database (Denmark)

    Frydendahl, K.; Imberechts, H.; Lehmann, S.

    2001-01-01

    (STa, STb, EAST1) and heat labile LT) enterotoxins and the verocytotoxin variant 2e (VT2e). To correctly identify false negative results, an endogenous internal control targeting the E. coil 16S rRNA gene was incorporated in each test tube. The assay was evaluated using a collection of E. coil...... reference strains which have previously been examined with phenotypical assays or DNA hybridization. Furthermore, the assay was evaluated by testing porcine E. coil field strains, previously characterized. The 5' nuclease assay correctly detected the presence of virulence genes in all reference strains....... When testing field strains there was generally excellent agreement with results obtained by laboratories in Belgium and Germany. In conclusion, the 5' nuclease assay developed is a fast and specific tool for detection of E. coli virulence genes in the veterinary diagnostic laboratory....

  15. In situ hybridization to detect porcine reproductive and respiratory syndrome virus

    DEFF Research Database (Denmark)

    Novosel, Dinko; Hjulsager, Charlotte Kristiane; Larsen, Lars Erik

    2012-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has for nearly 3 decades been economically one of the most important swine diseases. Despite intensive research focus, many unanswered questions remain regarding the pathogenesis of PRRSV. In situ hybridization (ISH) is generally considered...

  16. Bioinformatics prediction of swine MHC class I epitopes from Porcine Reproductive and Respiratory Syndrome Virus

    DEFF Research Database (Denmark)

    Welner, Simon; Nielsen, Morten; Lund, Ole

    Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) causes one of the most important diseases in all swine producing countries. The infection has a high impact on animal welfare, food safety and production economics. PRRSV possesses multiple immunoevasive strategies, from suppression...

  17. Novel insights into host responses and the reproductive pathophysiology of type 2 porcine reproductive and respiratory syndrome (PRRS)

    Science.gov (United States)

    A large-scale challenge experiment using type 2 porcine reproductive and respiratory virus (PRRSV) provided new insights into the pathophysiology of reproductive PRRS in third-trimester pregnant gilts. Deep phenotyping enabled identification of maternal and fetal factors predictive of PRRS severity ...

  18. Genomic prediction of piglet response to infection with one of two porcine reproductive and respiratory syndrome virus isolates.

    Science.gov (United States)

    Waide, Emily H; Tuggle, Christopher K; Serão, Nick V L; Schroyen, Martine; Hess, Andrew; Rowland, Raymond R R; Lunney, Joan K; Plastow, Graham; Dekkers, Jack C M

    2018-02-01

    Genomic prediction of the pig's response to the porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) would be a useful tool in the swine industry. This study investigated the accuracy of genomic prediction based on porcine SNP60 Beadchip data using training and validation datasets from populations with different genetic backgrounds that were challenged with different PRRSV isolates. Genomic prediction accuracy averaged 0.34 for viral load (VL) and 0.23 for weight gain (WG) following experimental PRRSV challenge, which demonstrates that genomic selection could be used to improve response to PRRSV infection. Training on WG data during infection with a less virulent PRRSV, KS06, resulted in poor accuracy of prediction for WG during infection with a more virulent PRRSV, NVSL. Inclusion of single nucleotide polymorphisms (SNPs) that are in linkage disequilibrium with a major quantitative trait locus (QTL) on chromosome 4 was vital for accurate prediction of VL. Overall, SNPs that were significantly associated with either trait in single SNP genome-wide association analysis were unable to predict the phenotypes with an accuracy as high as that obtained by using all genotyped SNPs across the genome. Inclusion of data from close relatives into the training population increased whole genome prediction accuracy by 33% for VL and by 37% for WG but did not affect the accuracy of prediction when using only SNPs in the major QTL region. Results show that genomic prediction of response to PRRSV infection is moderately accurate and, when using all SNPs on the porcine SNP60 Beadchip, is not very sensitive to differences in virulence of the PRRSV in training and validation populations. Including close relatives in the training population increased prediction accuracy when using the whole genome or SNPs other than those near a major QTL.

  19. Reactomes of porcine alveolar macrophages infected with porcine reproductive and respiratory syndrome virus.

    Directory of Open Access Journals (Sweden)

    Zhihua Jiang

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS has devastated pig industries worldwide for many years. It is caused by a small RNA virus (PRRSV, which targets almost exclusively pig monocytes or macrophages. In the present study, five SAGE (serial analysis of gene expression libraries derived from 0 hour mock-infected and 6, 12, 16 and 24 hours PRRSV-infected porcine alveolar macrophages (PAMs produced a total 643,255 sequenced tags with 91,807 unique tags. Differentially expressed (DE tags were then detected using the Bayesian framework followed by gene/mRNA assignment, arbitrary selection and manual annotation, which determined 699 DE genes for reactome analysis. The DAVID, KEGG and REACTOME databases assigned 573 of the DE genes into six biological systems, 60 functional categories and 504 pathways. The six systems are: cellular processes, genetic information processing, environmental information processing, metabolism, organismal systems and human diseases as defined by KEGG with modification. Self-organizing map (SOM analysis further grouped these 699 DE genes into ten clusters, reflecting their expression trends along these five time points. Based on the number one functional category in each system, cell growth and death, transcription processes, signal transductions, energy metabolism, immune system and infectious diseases formed the major reactomes of PAMs responding to PRRSV infection. Our investigation also focused on dominant pathways that had at least 20 DE genes identified, multi-pathway genes that were involved in 10 or more pathways and exclusively-expressed genes that were included in one system. Overall, our present study reported a large set of DE genes, compiled a comprehensive coverage of pathways, and revealed system-based reactomes of PAMs infected with PRRSV. We believe that our reactome data provides new insight into molecular mechanisms involved in host genetic complexity of antiviral activities against PRRSV and

  20. Monkey Viperin Restricts Porcine Reproductive and Respiratory Syndrome Virus Replication.

    Directory of Open Access Journals (Sweden)

    Jianyu Fang

    Full Text Available Porcine reproductive and respiratory syndrome virus (PRRSV is an important pathogen which causes huge economic damage globally in the swine industry. Current vaccination strategies provide only limited protection against PRRSV infection. Viperin is an interferon (IFN stimulated protein that inhibits some virus infections via IFN-dependent or IFN-independent pathways. However, the role of viperin in PRRSV infection is not well understood. In this study, we cloned the full-length monkey viperin (mViperin complementary DNA (cDNA from IFN-α-treated African green monkey Marc-145 cells. It was found that the mViperin is up-regulated following PRRSV infection in Marc-145 cells along with elevated IRF-1 gene levels. IFN-α induced mViperin expression in a dose- and time-dependent manner and strongly inhibits PRRSV replication in Marc-145 cells. Overexpression of mViperin suppresses PRRSV replication by blocking the early steps of PRRSV entry and genome replication and translation but not inhibiting assembly and release. And mViperin co-localized with PRRSV GP5 and N protein, but only interacted with N protein in distinct cytoplasmic loci. Furthermore, it was found that the 13-16 amino acids of mViperin were essential for inhibiting PRRSV replication, by disrupting the distribution of mViperin protein from the granular distribution to a homogeneous distribution in the cytoplasm. These results could be helpful in the future development of novel antiviral therapies against PRRSV infection.

  1. Gammadelta lymphocyte response to porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Olin, Michael R; Batista, Laura; Xiao, Zhengguo; Dee, Scott A; Murtaugh, Michael P; Pijoan, Carlos C; Molitor, Thomas W

    2005-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be one of the most important diseases facing swine industry today. Following PRRSV infection pigs develop both humoral and cell-mediated responses following PRRSV exposure; however, the relative importance in protection and clearance of the virus is not yet completely understood. Swine contain a large percentage of gammadelta T-lymphocytes in peripheral circulation capable of responding to various pathogens in both an innate and specific immune response. The objectives of this study were to determine whether gammadelta lymphocytes functionally respond to PRRSV upon initial exposure and re-exposure. Four month old PRRSV free gilts were intranasally inoculated with a field isolate MN-30100 then assessed at various time points post infection. On day 120, pigs were re-exposed with MN-30100 PRRSV strain and subsequently were bled on days 0, 7, and 14 post re-exposure. Lymphocyte subpopulations, antigen specific proliferation, and IFN-gamma production were evaluated throughout the study. Circulating gammadelta lymphocytes in PRRSV exposed animals expanded between days 14 to 70 (d14-d70, p = 0.016); following antigen stimulation, gammadelta lymphocyte proliferated by day 14 (d0-d14, p = 0.001) continuing through day 60. gammadelta lymphocytes produced IFN-gamma by day 14 pi continuing through day 50 (d0-d50, p = 0.004). Following re-exposure both gammadelta+ and CD4+ lymphocytes increased in IFN-gamma production. These results are not fully conclusive on the role of gammadelta lymphocytes against PRRSV; the data indicate that gammadelta lymphocytes specifically respond to PRRSV.

  2. Seroprevalence of porcine reproductive and respiratory syndrome, Aujeszky's disease, and porcine parvovirus in replacement gilts in Thailand.

    Science.gov (United States)

    Tummaruk, Padet; Tantilertcharoen, Rachod

    2012-06-01

    The present study investigated the seroprevalence of porcine reproductive and respiratory syndrome virus, Aujeszky's disease virus (ADV), and porcine parvovirus (PPV) in replacement gilts from selected five swine herds in Thailand. The study consisted of three parts. First, a retrospective data analysis on the seroprevalence of porcine reproductive and respiratory syndrome virus (PRRSV) and ADV glycoprotein I (gI) in gilts, sows, boars, nursery, and fattening pigs in five herds (n = 7,030). Second, a cross-sectional study on seroprevalence of PRRSV, ADV, and PPV (n = 200) in replacement gilts. Last, the seroprevalence of PRRSV, ADV, and PPV in gilts culled due to reproductive failure (n = 166). Across the herds, the seroprevalence of PRRSV and ADV was 79.3% and 5.3%, respectively. The cross-sectional study revealed that 87.5%, 4.0%, and 99.0% of the replacement gilts were infected with PRRSV, ADV, and PPV, respectively. In the gilts culled due to reproductive failure, the seroprevalence of PRRSV, ADV, and PPV was 73.5%, 28.3%, and 86.0%, respectively. Of these culled gilts, 75.5% had been infected with at least two viruses and 18.9% had been infected with all three viruses. It could be concluded that most of the replacement gilts were exposed to PRRSV (84%), PPV (97%), and ADV (4%) before entering the breeding house. PPV was an enzootic disease among the selected herds. The prevalence of ADV was higher in gilts culled due to reproductive disturbance than in the healthy gilts.

  3. Molecular characterization of porcine circovirus 2 isolated from diseased pigs co-infected with porcine reproductive and respiratory syndrome virus

    Directory of Open Access Journals (Sweden)

    Liu Chengqian

    2010-10-01

    Full Text Available Abstract In this study, we isolated a porcine circovirus 2 (PCV2 strain from piglets co-infected with porcine reproductive and respiratory syndrome virus (PRRSV. The complete genome of this strain was sequenced, phylogenetic and polymorphic analyses were carried out. BLAST searches revealed the highest sequence identity (99.5% nt and 99.3% aa to Guangxi strain EF675230. The phylogenetic tree showed that clustering of the isolates didn't strongly correlate to geographical distribution. Polymorphic analyses demonstrated that the amino acids at most of the polymorphic sites in Open Reading Frame 1(ORF1 and 2 (ORF2belong to the same amino acid group according to chemical or structural properties, and revealed that highly polymorphic regions overlapped with the known immunoreactive epitopes of ORF2.

  4. Porcine respiratory disease complex: Interaction of vaccination and porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, and Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Chae, Chanhee

    2016-06-01

    Porcine respiratory disease is a multifactorial and complex disease caused by a combination of infectious pathogens, environmental stressors, differences in production systems, and various management practices; hence the name porcine respiratory disease complex (PRDC) is used. Porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), and Mycoplasma hyopneumoniae are considered to be the most important pathogens that cause PRDC. Although interactions among the three major respiratory pathogens are well documented, it is also necessary to understand the interaction between vaccines and the three major respiratory pathogens. PRRSV and M. hyopneumoniae are well known to potentiate PCV2-associated lesions; however, PRRSV and mycoplasmal vaccines can both enhance PCV2 viraemia regardless of the effects of the actual PRRSV or M. hyopneumoniae infection. On the other hand, M. hyopneumoniae potentiates the severity of pneumonia induced by PRRSV, and vaccination against M. hyopneumoniae alone is also able to decrease PRRSV viraemia and PRRSV-induced lung lesions in dually infected pigs. This review focuses on (1) interactions between PCV2, PRRSV, and M. hyopneumoniae; and (2) interactions between vaccines and the three major respiratory pathogens. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus

    NARCIS (Netherlands)

    Wissink, E.H.J.; Kroese, M.V.; Wijk, van H.A.; Rijsewijk, F.A.M.; Meulenberg, J.J.; Rottier, P.J.M.

    2005-01-01

    Virions of porcine reproductive and respiratory syndrome virus (PRRSV) contain six membrane proteins: the major proteins GP5 and M and the minor proteins GP2a, E, GP3, and GP4. Here, we studied the envelope protein requirements for PRRSV particle formation and infectivity using full-length cDNA

  6. Porcine Respiratory and Reproductive Syndrome Virus Variants, Vietnam and China, 2007

    OpenAIRE

    Feng, Youjun; Zhao, Tiezhu; Nguyen, Tung; Inui, Ken; Ma, Ying; Nguyen, Thi Hoa; Nguyen, Cam; Liu, Di; Bui, Quang Anh; To, Long Thanh; Wang, Chuanbin; Tian, Kegong; Gao, George F.

    2008-01-01

    We characterized isolates from porcine respiratory and reproductive syndrome virus epidemics in Vietnam and China in 2007. These isolates showed ?99% identity at the genomic level. Genetic analysis indicated that they share a discontinuous deletion of 30 aa in nonstructural protein 2, which indicates that identical variants emerged in Vietnam and China.

  7. Porcine respiratory and reproductive syndrome virus variants, Vietnam and China, 2007.

    Science.gov (United States)

    Feng, Youjun; Zhao, Tiezhu; Nguyen, Tung; Inui, Ken; Ma, Ying; Nguyen, Thi Hoa; Nguyen, Van Cam; Liu, Di; Bui, Quang Anh; To, Long Thanh; Wang, Chuanbin; Tian, Kegong; Gao, George F

    2008-11-01

    We characterized isolates from porcine respiratory and reproductive syndrome virus epidemics in Vietnam and China in 2007. These isolates showed approximately 99% identity at the genomic level. Genetic analysis indicated that they share a discontinuous deletion of 30 aa in nonstructural protein 2, which indicates that identical variants emerged in Vietnam and China.

  8. Porcine Reproductive and Respiratory Syndrome (PRRS) with special reference to clinical aspects and diagnosis: a review

    NARCIS (Netherlands)

    Nodelijk, G.

    2002-01-01

    After a short introduction on Porcine Reproductive and Respiratory Syndrome (PRRS) regarding the history, the first occurrence in several countries, and the causal virus, designated Lelystad virus, a description is given of the clinical aspects and several diagnostic methods. After some general

  9. Genetic control of host resistance to porcine reproductive and respiratory syndrome virus (PRRSV) infection

    Science.gov (United States)

    This manuscript focuses on the advances made using genomic approaches to identify biomarkers that define genes and pathways that are correlated with swine resistance to infection with porcine reproductive and respiratory syndrome virus (PRRSV), the most economically important swine viral pathogen wo...

  10. Inactivated and subunit vaccines against porcine reproductive and respiratory syndrome: Current status and future direction.

    Science.gov (United States)

    Renukaradhya, Gourapura J; Meng, Xiang-Jin; Calvert, Jay G; Roof, Michael; Lager, Kelly M

    2015-06-17

    Within a few years of its emergence in the late 1980s, the PRRS virus had spread globally to become the foremost infectious disease concern for the pork industry. Since 1994, modified live-attenuated vaccines against porcine reproductive and respiratory syndrome virus (PRRSV-MLV) have been widely used, but have failed to provide complete protection against emerging and heterologous field strains of the virus. Moreover, like many other MLVs, PRRSV-MLVs have safety concerns including vertical and horizontal transmission of the vaccine virus and several documented incidences of reversion to virulence. Thus, the development of efficacious inactivated vaccines is warranted for the control and eradication of PRRS. Since the early 1990s, researchers have been attempting to develop inactivated PRRSV vaccines, but most of the candidates have failed to elicit protective immunity even against homologous virus challenge. Recent research findings relating to both inactivated and subunit candidate PRRSV vaccines have shown promise, but they need to be pursued further to improve their heterologous efficacy and cost-effectiveness before considering commercialization. In this comprehensive review, we provide information on attempts to develop PRRSV inactivated and subunit vaccines. These includes various virus inactivation strategies, adjuvants, nanoparticle-based vaccine delivery systems, DNA vaccines, and recombinant subunit vaccines produced using baculovirus, plant, and replication-deficient viruses as vector vaccines. Finally, future directions for the development of innovative non-infectious PRRSV vaccines are suggested. Undoubtedly there remains a need for novel PRRSV vaccine strategies targeted to deliver cross-protective, non-infectious vaccines for the control and eradication of PRRS. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Innate and adaptive immunity against Porcine Reproductive and Respiratory Syndrome Virus.

    Science.gov (United States)

    Loving, Crystal L; Osorio, Fernando A; Murtaugh, Michael P; Zuckermann, Federico A

    2015-09-15

    Many highly effective vaccines have been produced against viruses whose virulent infection elicits strong and durable protective immunity. In these cases, characterization of immune effector mechanisms and identification of protective epitopes/immunogens has been informative for the development of successful vaccine programs. Diseases in which the immune system does not rapidly clear the acute infection and/or convalescent immunity does not provide highly effective protection against secondary challenge pose a major hurdle for clinicians and scientists. Porcine reproductive and respiratory syndrome virus (PRRSV) falls primarily into this category, though not entirely. PRRSV causes a prolonged infection, though the host eventually clears the virus. Neutralizing antibodies can provide passive protection when present prior to challenge, though infection can be controlled in the absence of detectable neutralizing antibodies. In addition, primed pigs (through natural exposure or vaccination with a modified-live vaccine) show some protection against secondary challenge. While peripheral PRRSV-specific T cell responses have been examined, their direct contribution to antibody-mediated immunity and viral clearance have not been fully elucidated. The innate immune response following PRRSV infection, particularly the antiviral type I interferon response, is meager, but when provided exogenously, IFN-α enhances PRRSV immunity and viral control. Overall, the quality of immunity induced by natural PRRSV infection is not ideal for informing vaccine development programs. The epitopes necessary for protection may be identified through natural exposure or modified-live vaccines and subsequently applied to vaccine delivery platforms to accelerate induction of protective immunity following vaccination. Collectively, further work to identify protective B and T cell epitopes and mechanisms by which PRRSV eludes innate immunity will enhance our ability to develop more effective methods

  12. Fresh Pork and Porcine Reproductive and Respiratory Syndrome Virus: Factors Related to the Risk of Disease Transmission.

    Science.gov (United States)

    Hall, W; Neumann, E

    2015-08-01

    Porcine reproductive and respiratory syndrome virus (PRRS) is a highly infectious virus. Experimentally, the disease can be induced in naïve pigs by the oral, intranasal and intramuscular routes. Depending on the virulence of the strain of the virus and the age of the pig, peak viremia can occur within 7 days of infection, and live virus can be isolated from blood or lymph nodes for several months post-infection. Young pigs tend to develop higher titres of viremia than older pigs infected by the same route and dose with the same strain of virus. Porcine reproductive and respiratory syndrome virus survives in pork harvested from infected pigs for extended periods at temperatures of -20 or -70°C. In experimentally infected pigs, survival of PRRS virus in muscle held at 4°C has been demonstrated for at least 7 days, and infectivity of the virus in these samples was confirmed by bioassay. The optimal pH range for the survival of PRRS virus is thought to be 6.0 to 7.5. The elevated pH of non-meat tissues (generally one pH unit higher) is likely to favour extended survival of PRRS virus in pig carcasses from which all superficial and deep lymph nodes have not been removed. It is likely that exsanguinated carcasses held at 4°C retain sufficient blood or lymph tissue to contain infective doses of PRRS virus. Porcine reproductive and respiratory syndrome virus is rapidly inactivated by heat, providing a predictable method to ensure that pork tissues are free of viable virus and feeding of cooked swill or garbage should not constitute a risk to pigs. While the probability of viable PRRS virus being present in a pig carcass may be low, the risk is not zero. The importation of raw pork into countries where PRRS is not endemic represents a hazard with potentially severe economic consequences. © 2013 Blackwell Verlag GmbH.

  13. Pathogenicity of three type 2 Porcine Reproductive and Respiratory Syndrome virus strains in experimentally inoculated pregnant gilts

    Science.gov (United States)

    Mechanisms of reproductive failure resulting from infection with porcine reproductive and respiratory syndrome virus (PRRSv) are still poorly understood. The present study, a side-by-side evaluation of the pathogenicity of three type 2 PRRSv strains in a reproductive model, was used as a pilot study...

  14. Porcine reproductive and respiratory syndrome virus nonstructural protein 2 (nsp2) topology and selective isoform integration in artificial membranes

    Science.gov (United States)

    Membrane modification of host subcellular compartments is critical to the replication of many RNA viruses. Enveloped viruses additionally require the ability to requisition cellular membranes during egress for the development of infectious progeny. Porcine reproductive and respiratory syndrome virus...

  15. Evaluation of systems for reducing the transmission of Porcine reproductive and respiratory syndrome virus by aerosol

    OpenAIRE

    Dee, Scott A.; Batista, Laura; Deen, John; Pijoan, Carlos

    2006-01-01

    The purpose of this study was to compare 3 methods for the reduction of aerosol transmission of Porcine reproductive and respiratory syndrome virus (PRRSV): high-efficiency particulate air (HEPA) filtration, low-cost filtration, and ultraviolet light (UV) irradiation. The HEPA-filtration system involved a pre-filter screen, a bag filter (EU8 rating), and a HEPA filter (EU13 rating). The low-cost-filtration system contained mosquito netting (pre-filter), a fiberglass furnace filter, and an ele...

  16. Porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 infections in wild boar (Sus scrofa) in southwestern Germany.

    Science.gov (United States)

    Hammer, Ralf; Ritzmann, Mathias; Palzer, Andreas; Lang, Christiane; Hammer, Birgit; Pesch, Stefan; Ladinig, Andrea

    2012-01-01

    Samples were collected from 203 wild boars (Sus scrofa) hunted in Baden-Wurtemburg, Germany from November-January 2008 and 2009. Samples from the lung and tonsil were analyzed by quantitative polymerase chain reaction (qPCR) for porcine reproductive and respiratory syndrome virus (PRRSV) type 1 (European type) and type 2 (American type). A qPCR to detect porcine circovirus type 2 (PCV2)-specific genome was performed on tissue homogenates including lung, tonsils, and inguinal lymph nodes. Serum samples were tested for antibodies against PRRSV and PCV2 by enzyme-linked immunosorbent assay (ELISA). No PRRSV was detected in any of the 203 samples and one sample had detectable antibodies against PRRSV. We detected PCV2 in organ materials from 103 wild boars with a prevalence of 50.7%. The number of wild boars positive for PCV2 by PCR varied according to the population density of wild boars among woodlands. More positive samples were detected in woodlands with a high density of wild boars. We found no correlation between the number of PCV2-positive wild boars and the density of domestic pigs in the surrounding area. The number of wild boars positive for antibodies against PCV2 by the INGEZIM Circovirus IgG/IgM test kit was low (53 sera positive for IgG- and three sera positive for IgM-antibodies) in comparison to the higher positive results from the INGEZIM CIRCO IgG test kit (102 positive and 12 inconclusive results).

  17. Evaluation of a modified live-virus vaccine for the prevention of porcine parvovirus-induced reproductive disease in swine.

    Science.gov (United States)

    Paul, P S; Mengeling, W L

    1980-12-01

    Each of 5 gilts was vaccinated IM with modified live-virus (MLV) vaccine for porcine parvovirus (PPV), and 5 gilts were used as nonvaccinated controls. Vaccinated gilts developed hemagglutination-inhibiting (HI) antibodies to PPV (titer of 320 to 1,280) by 2 weeks after vaccination. All gilts wee bred, and at about 40 days of gestation their immunity was challenged by intranasal and oral administration of a virulent strain of PPV. Gilts were killed at about 84 days of gestation and their litters were examined. Litters from vaccinated gilts comprised 59 live and 2 dead fetuses. Neither the virus nor antibodies to the virus were detected in any of the fetuses. In contrast, litters from nonvaccinated gilts comprised 25 live and 29 dead fetuses, and PPV was isolated from all dead and 9 live fetuses. Viral antigen was detected by direct immunofluorescence in the lungs of all PPV-infected fetuses, and 7 of the live infected fetuses had HI antibodies. Modified live virus did not cause transplacental fetal infection in 3 seronegative gilts inoculated at about 40 days of gestation. All 3 gilts had antibodies to PPV (80 to 640) when killed at about 84 days of gestation, and neither HI antibodies nor PPV were detected in any of the 20 live and 1 dead fetuses from these gilts. Modified live virus replicated in the tissues of fetuses of 2 gilts inoculated in utero. Seven of 8 fetuses inoculated with MLV died and were infected. These results demonstrated the efficacy of MLV vaccine for the prevention of PPV-induced reproductive disease in swine. Although MLV did not cross the placental barrier in pregnant animals, its pathogenicity for porcine fetuses after direct in utero inoculation indicates that its use should be limited to nonpregnant animals.

  18. Antibiotic resistance, virulence determinants and production of biogenic amines among enterococci from ovine, feline, canine, porcine and human milk.

    Science.gov (United States)

    Jiménez, Esther; Ladero, Victor; Chico, Irene; Maldonado-Barragán, Antonio; López, María; Martín, Virginia; Fernández, Leonides; Fernández, María; Álvarez, Miguel A; Torres, Carmen; Rodríguez, Juan M

    2013-12-10

    Recent studies have shown that mammalian milk represents a continuous supply of commensal bacteria, including enterococci. The objectives of this study were to evaluate the presence of enterococci in milk of different species and to screen them for several genetic and phenotypic traits of clinical significance among enterococci. Samples were obtained from, at least, nine porcine, canine, ovine, feline and human healthy hosts. Enterococci could be isolated, at a concentration of 1.00 × 10(2) -1.16 × 10(3) CFU/ml, from all the porcine samples and, also from 85, 50, 25 and 25% of the human, canine, feline and ovine ones, respectively. They were identified as Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Enterococcus casseliflavus and Enterococcus durans. Among the 120 initial enterococcal isolates, 36 were selected on the basis of their different PFGE profiles and further characterized. MLST analysis revealed a wide diversity of STs among the E. faecalis and E. faecium strains, including some frequently associated to hospital infections and novel STs. All the E. faecalis strains possessed some of the potential virulence determinants (cad, ccf, cob, cpd, efaA(fs), agg2, gelE, cylA, esp(fs)) assayed while the E. faecium ones only harboured the efaA(fm) gene. All the tested strains were susceptible to tigecycline, linezolid and vancomycin, and produced tyramine. Their susceptibility to the rest of the antimicrobials and their ability to produce other biogenic amines varied depending on the strain. Enterococci strains isolated from porcine samples showed the widest spectrum of antibiotic resistance. Enterococci isolated from milk of different mammals showed a great genetic diversity. The wide distribution of virulence genes and/or antibiotic resistance among the E. faecalis and E. faecium isolates indicates that they can constitute a reservoir of such traits and a risk to animal and human health.

  19. Differential cellular protein expression in continuous porcine alveolar macrophages regulated by the porcine reproductive and respiratory syndrome virus nucleocapsid protein.

    Science.gov (United States)

    Sagong, Mingeun; Lee, Changhee

    2010-07-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a leading cause of significant economic losses in the pig industry worldwide. PRRSV infects preferentially porcine alveolar macrophages (PAMs) and subsequently utilizes the host cell biosynthetic machinery for its own replication. To date, a number of studies have been conducted to investigate compensatory changes of cellular gene expression of PAMs upon PRRSV infection. However, very little information exists about differential cellular protein expression of the natural target cells regulated by each viral protein. This study was therefore designed to examine the dynamics of host protein expression of continuous PAM cells by the PRRSV nucleocapsid (N) protein that is the most abundant and multifunctional viral component. We first established sublines of PAM cells to stably express the PRRSV N protein and assessed alterations in cellular protein productions of N-expressing PAM (PAM-pCD163-N) cells at different time courses by the use of proteomic analysis. A total of 23 protein spots were initially found to be differentially expressed in PAM-pCD163-N cells compared with normal PAM cells by high-resolution two-dimensional gel electrophoresis (2DE). Of these spots, 15 protein spots with statistically significant alteration, including 4 up-regulated and 11 down-regulated protein spots, were picked out for subsequent protein identification by peptide mass fingerprinting after matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS). The altered cellular proteins identified in this study were classified into the functions involved in a variety of cellular processes such as cell division, metabolism, inflammation response, stress response, ubiquitin-proteasome pathway, protein folding and synthesis, and transportation. Notably, heat shock 27kDa protein (HSP27) was found to be up-regulated in PAM-pCD163-N cells. The proteomics data will provide insights into the specific

  20. Efficacy of an inactivated virus vaccine for prevention of porcine parvovirus-induced reproductive failure.

    Science.gov (United States)

    Mengeling, W L; Brown, T T; Paul, P S; Gutekunst, D E

    1979-02-01

    Gilts vaccinated IM either once (4 gilts) or twice (2 gilts) with an acetylethyleneimine-inactivated porcine parvovirus (PPV) vaccine before they were bred were subsequently exposed intranasally and orally to virulent PPV at about the 40th day of gestation (from 37 to 43 days). At 2 weeks after vaccination, all had hemagglutination-inhibiting (HI) titers for PPV (from 20 to 80) which decreased by the time the immunity was challenged with virulent virus (from 10 to 40), but increased thereafter (from 160 to 1,280). Titers of singly and doubly vaccinated gilts were similar throughout the experiment. The gilts were killed at about the 84th day of gestation (from 80 to 87 days), and their litters were examined. Litters were comprised of 68 live fetuses and 1 dead fetus (7 to 14 fetuses/litter). Neither viral antigen, PPV, nor homologous HI antibody was found in any of the fetuses. In addition, 4 gilts were kept in contact with the vaccinated gilts and were treated similarly except for vaccination. These 4 gilts remained free of HI antibody until after they were exposed to virulent PPV during gestation. At the time the gilts were killed the titers were 1,280 to 2,560. Their litters were comprised of 11 live fetuses and 26 dead fetuses (8 to 11 fetuses/litter). Virus was isolated from fetuses of all litters. Viral antigen was found in 24 of the dead fetuses and 10 of the live fetuses. All infected live fetuses also had HI antibody for PPV. The 2 boars used to breed vaccinated and nonvaccinated gilts (usually each gilt was bred to each of the 2 boars), but not exposed to virulent PPV, remained free of HI antibody for PPV.

  1. Porcine reproductive and respiratory syndrome virus infection triggers HMGB1 release to promote inflammatory cytokine production.

    Science.gov (United States)

    Duan, Erzhen; Wang, Dang; Luo, Rui; Luo, Jingyi; Gao, Li; Chen, Huanchun; Fang, Liurong; Xiao, Shaobo

    2014-11-01

    The high mobility group box 1 (HMGB1) protein is an endogenous damage-associated molecular pattern (DAMP) molecule involved in the pathogenesis of various infectious agents. Based on meta-analysis of all publicly available microarray datasets, HMGB1 has recently been proposed as the most significant immune modulator during the porcine response to porcine reproductive and respiratory syndrome virus (PRRSV) infection. However, the function of HMGB1 in PRRSV pathogenesis is unclear. In this study, we found that PRRSV infection triggers the translocation of HMGB1 from the nucleus to the extracellular milieu in MARC-145 cells and porcine alveolar macrophages. Although HMGB1 has no effect on PRRSV replication, HMGB1 promotes PRRSV-induced NF-κB activation and subsequent expression of inflammatory cytokines through receptors RAGE, TLR2 and TLR4. Our findings show that HMGB1 release, triggered by PRRSV infection, enhances the efficiency of virus-induced inflammatory responses, thereby providing new insights into the pathogenesis of PRRSV infection. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Porcine reproductive and respiratory syndrome virus infection triggers HMGB1 release to promote inflammatory cytokine production

    Energy Technology Data Exchange (ETDEWEB)

    Duan, Erzhen; Wang, Dang; Luo, Rui; Luo, Jingyi; Gao, Li; Chen, Huanchun; Fang, Liurong, E-mail: fanglr@mail.hzau.edu.cn; Xiao, Shaobo, E-mail: vet@mail.hzau.edu.cn

    2014-11-15

    The high mobility group box 1 (HMGB1) protein is an endogenous damage-associated molecular pattern (DAMP) molecule involved in the pathogenesis of various infectious agents. Based on meta-analysis of all publicly available microarray datasets, HMGB1 has recently been proposed as the most significant immune modulator during the porcine response to porcine reproductive and respiratory syndrome virus (PRRSV) infection. However, the function of HMGB1 in PRRSV pathogenesis is unclear. In this study, we found that PRRSV infection triggers the translocation of HMGB1 from the nucleus to the extracellular milieu in MARC-145 cells and porcine alveolar macrophages. Although HMGB1 has no effect on PRRSV replication, HMGB1 promotes PRRSV-induced NF-κB activation and subsequent expression of inflammatory cytokines through receptors RAGE, TLR2 and TLR4. Our findings show that HMGB1 release, triggered by PRRSV infection, enhances the efficiency of virus-induced inflammatory responses, thereby providing new insights into the pathogenesis of PRRSV infection. - Highlights: • PRRSV infection triggers HMGB1 release from MARC-145 cells and PAMs. • HMGB1 does not significantly affect PRRSV proliferation. • HMGB1 is involved in PRRSV-induced NF-κB activation and inflammatory responses. • HMGB1 promotes PRRSV-induced inflammatory responses through TLR2/4 and RAGE.

  3. Effect of porcine reproductive and respiratory syndrome virus (PRRSV) on alveolar lung macrophage survival and function

    DEFF Research Database (Denmark)

    Oleksiewicz, Martin B.; Nielsen, Jens

    1999-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) recently emerged as an important cause of reproductive disorders and pneumonia in domestic pigs throughout the world. Acute cytocidal replication of PRRSV in alveolar lung macrophages causes the acute pneumonia; however, it remains largely....... Monitored by flow cytometric detection of intracellular PRRSV nucleocapsid protein, acute (24 h post infection) PRRSV replication did not impede the ability of alveolar macrophages to ingest fluorescently labelled Escherichia coli. At 48 h post infection, PRRSV-induced cytotoxicity (quantitated by flow...... analysis of cell size and membrane integrity) led to 40% reduction in the total number of phagocytozing cells. However, viable/uninfected macrophages in PRRSV-infected cultures exhibited normal phagocytic ability at 48 h, indicating that no soluble phagocytosis-suppressive mediators were induced by PRRSV...

  4. Characterization of the microRNAome in porcine reproductive and respiratory syndrome virus infected macrophages.

    Directory of Open Access Journals (Sweden)

    Julie A Hicks

    Full Text Available Porcine Reproductive and Respiratory Syndrome Virus (PRRSV, a member of the arterivirus family, is the causative agent of Porcine Reproductive and Respiratory Syndrome (PRRS. PRRS is characterized by late term abortions and respiratory disease, particularly in young pigs. Small regulatory RNAs termed microRNA (miRNA are associated with gene regulation at the post-transcriptional level. MiRNAs are known to play many diverse and complex roles in viral infections. To discover the impact of PRRSV infections on the cellular miRNAome, Illumina deep sequencing was used to construct small RNA expression profiles from in vitro cultured PRRSV-infected porcine alveolar macrophages (PAMs. A total of forty cellular miRNAs were significantly differentially expressed within the first 48 hours post infection (hpi. The expression of six miRNAs, miR-30a-3p, miR-132, miR-27b*, miR-29b, miR-146a and miR-9-2, were altered at more than one time point. Target gene identification suggests that these miRNAs are involved in regulating immune signaling pathways, cytokine, and transcription factor production. The most highly repressed miRNA at 24 hpi was miR-147. A miR-147 mimic was utilized to maintain miR-147 levels in PRRSV-infected PAMs. PRRSV replication was negatively impacted by high levels of miR-147. Whether down-regulation of miR-147 is directly induced by PRRSV or if it is part of the cellular response and PRRSV indirectly benefits remains to be determined. No evidence could be found of PRRSV-encoded miRNAs. Overall, the present study has revealed that a large and diverse group of miRNAs are expressed in swine alveolar macrophages and that the expression of a subset of these miRNAs is altered in PRRSV infected macrophages.

  5. Risk factors for infection of sow herds with porcine reproductive and respiratory syndrome (PRRS) virus

    DEFF Research Database (Denmark)

    Mortensen, Sten; Stryhn, Henrik; Søgaard, Rikke

    2002-01-01

    In 1992, the porcine reproductive and respiratory syndrome virus (PRRSV) of European type (PRRSV-EU) was introduced in Denmark. By 1996, the virus had spread to approximately 25% of the Danish herds. In January 1996, a modified-live vaccine based on the American type of the virus (PRRSV-US) was u......In 1992, the porcine reproductive and respiratory syndrome virus (PRRSV) of European type (PRRSV-EU) was introduced in Denmark. By 1996, the virus had spread to approximately 25% of the Danish herds. In January 1996, a modified-live vaccine based on the American type of the virus (PRRSV......-US) was used in replacement boars for Danish artificial insemination (AI) centres and from July 1996, the vaccine was used in PRRSV-EU infected herds for prevention of disease. Soon after vaccine introduction, PRRSV non-infected herds experienced outbreaks of disease due to infection with PRRSV......-US. In this study, we investigated the risk factors (biosecurity level, animals, exposure from PRRSV-US-infected neighbour herds, semen, herd size, pig density and herd density) for infection with PRRSV-US in a cohort of 1071 sow herds; we used a nested case-control study. The retrospective observation period...

  6. Virulence factors of Actinobacillus pleuropneumoniae involved in colonization, persistence and induction of lesions in its porcine host.

    Science.gov (United States)

    Chiers, Koen; De Waele, Tine; Pasmans, Frank; Ducatelle, Richard; Haesebrouck, Freddy

    2010-01-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia. The virulence factors of this microorganism involved in colonization and the induction of lung lesions have been thoroughly studied and some have been well characterized. A. pleuropneumoniae binds preferentially to cells of the lower respiratory tract in a process involving different adhesins and probably biofilm formation. Apx toxins and lipopolysaccharides exert pathogenic effects on several host cells, resulting in typical lung lesions. Lysis of host cells is essential for the bacterium to obtain nutrients from the environment and A. pleuropneumoniae has developed several uptake mechanisms for these nutrients. In addition to persistence in lung lesions, colonization of the upper respiratory tract--and of the tonsils in particular--may also be important for long-term persistent asymptomatic infection. Information on virulence factors involved in tonsillar and nasal cavity colonization and persistence is scarce, but it can be speculated that similar features as demonstrated for the lung may play a role. © The authors, published by INRA/EDP Sciences, 2010.

  7. iTRAQ-based comparative proteomic analysis of Vero cells infected with virulent and CV777 vaccine strain-like strains of porcine epidemic diarrhea virus.

    Science.gov (United States)

    Guo, Xiaozhen; Hu, Han; Chen, Fangzhou; Li, Zhonghua; Ye, Shiyi; Cheng, Shuang; Zhang, Mengjia; He, Qigai

    2016-01-01

    The re-emerging porcine epidemic diarrhea virus (PEDV) variant related diarrhea has been documented in China since late 2010 and now with global distribution. Currently, a virulent PEDV CH/YNKM-8/2013 and a CV777 vaccine strain-like AH-M have been successfully isolated from the clinical samples. To dissect out the underlying pathogenic mechanism of virulent PEDV and clarify the differences between virulent and CV777 vaccine strain-like PEDV infections, we performed an iTRAQ-based comparative quantitative proteomic study of Vero cells infected with both PEDV strains. A total of 661 and 474 differentially expressed proteins were identified upon virulent and CV777 vaccine strain-like isolates infection, respectively. Ingenuity Pathway Analysis was employed to investigate the canonical pathways and functional networks involved in both PEDV infections. Comprehensive studies have revealed that the PEDV virulent strain suppressed protein synthesis of Vero cells through down-regulating mTOR as well as its downstream targets 4EBP1 and p70S6K activities, which were validated by immunoblotting. In addition, the virulent strain could activate NF-κB pathway more intensively than the CV777 vaccine strain-like isolate, and elicit stronger inflammatory cascades as well. These data might provide new insights for elucidating the specific pathogenesis of PEDV infection, and pave the way for the development of effective therapeutic strategies. Porcine epidemic diarrhea is now worldwide distributed and causing huge economic losses to swine industry. The immunomodulation and pathogenesis between PEDV and host, as well as the difference between virulent and attenuated strains of PEDV infections are still largely unknown. In this study, we presented for the first application of proteomic analysis to compare whole cellular protein alterations induced by virulent and CV777 vaccine strain-like PEDV infections, which might contribute to understand the pathogenesis of PEDV and anti

  8. Sequence analysis of porcine reproductive and respiratory syndrome virus of the American type collected from Danish swine herds

    DEFF Research Database (Denmark)

    Madsen, K.G.; Hansen, C.M.; Madsen, E.S.

    1998-01-01

    Vaccine-like viruses of American type of porcine reproductive and respiratory syndrome virus (PRRSV) were detected in serum samples by RT-PCR. The viruses were analysed by nucleotide sequencing of the genomic region encoding open reading frames 2 to 7. During the ongoing study of Danish isolates...

  9. Safety and efficacy of a novel European vaccine for porcine reproductive and respiratory virus in bred gilts.

    Science.gov (United States)

    Piontkowski, Michael D; Kroll, Jeremy; Orveillon, Francois-Xavier; Kraft, Christian; Coll, Teresa

    2016-10-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) can be devastating to commercial breeding operations. The objective of this study was to evaluate a novel European PRRSV vaccinal strain for safety and efficacy in bred gilts. In 2 experiments, 110 gilts were vaccinated intramuscularly and the vaccine was evaluated for safety and efficacy. Gilts in Experiment 1 were evaluated for local and systemic reactions and gilts in both experiments were observed for clinical signs of disease through farrow. In both experiments, piglet clinical observations, piglet average daily weight gain (ADWG), gilt serology [determined by enzyme-linked immunosorbent assay (ELISA)], gilt and piglet viremia [determined by quantitative real-time polymerase chain reaction (qPCR)], as well as piglet lung lesion scores and PRRS virus in lung tissue (qPCR) were determined. The vaccine was shown to be safe as there were no significant differences among groups in either experiment. Efficacy was established in Experiment 2 as both vaccinated groups were associated with desirable significant differences in percentage of gilts with abnormal clinical findings; gilt viral load post-challenge [day 125, day of farrowing (DOF), and DOF + 13]; percentages of alive, healthy live, weak live, and mummified piglets per litter at farrowing and weaning; percentage of piglets per gilt that were positive for viremia; percentage of piglets per gilt with clinical disease; and piglet viral load on DOF. It was concluded that a vaccine formulated from the PRRSV modified live virus (MLV) strain 94881 is a safe and effective method of protection against the detrimental effects of virulent PRRSV infection in breeding female pigs.

  10. Pathogenicity and molecular characterization of emerging porcine reproductive and respiratory syndrome virus in Vietnam in 2007.

    Science.gov (United States)

    Metwally, S; Mohamed, F; Faaberg, K; Burrage, T; Prarat, M; Moran, K; Bracht, A; Mayr, G; Berninger, M; Koster, L; To, T L; Nguyen, V L; Reising, M; Landgraf, J; Cox, L; Lubroth, J; Carrillo, C

    2010-10-01

    In 2007, Vietnam experienced swine disease outbreaks causing clinical signs similar to the 'porcine high fever disease' that occurred in China during 2006. Analysis of diagnostic samples from the disease outbreaks in Vietnam identified porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV-2). Additionally, Escherichia coli and Streptococcus equi subspecies zooepidemicus were cultured from lung and spleen, and Streptococcus suis from one spleen sample. Genetic characterization of the Vietnamese PRRSV isolates revealed that this virus belongs to the North American genotype (type 2) with a high nucleotide identity to the recently reported Chinese strains. Amino acid sequence in the nsp2 region revealed 95.7-99.4% identity to Chinese strain HUN4, 68-69% identity to strain VR-2332 and 58-59% identity to strain MN184. A partial deletion in the nsp2 gene was detected; however, this deletion did not appear to enhance the virus pathogenicity in the inoculated pigs. Animal inoculation studies were conducted to determine the pathogenicity of PRRSV and to identify other possible agents present in the original specimens. Pigs inoculated with PRRSV alone and their contacts showed persistent fever, and two of five pigs developed cough, neurological signs and swollen joints. Necropsy examination showed mild to moderate bronchopneumonia, enlarged lymph nodes, fibrinous pericarditis and polyarthritis. PRRSV was re-isolated from blood and tissues of the inoculated and contact pigs. Pigs inoculated with lung and spleen tissue homogenates from sick pigs from Vietnam developed high fever, septicaemia, and died acutely within 72 h, while their contact pigs showed no clinical signs throughout the experiment. Streptococcus equi subspecies zooepidemicus was cultured, and PRRSV was re-isolated only from the inoculated pigs. Results suggest that the cause of the swine deaths in Vietnam is a multifactorial syndrome with PRRSV as a major factor. © 2010

  11. Role of CD151, A tetraspanin, in porcine reproductive and respiratory syndrome virus infection

    Directory of Open Access Journals (Sweden)

    Kapil Sanjay

    2007-06-01

    Full Text Available Abstract Background Porcine reproductive and respiratory syndrome virus (PRRSV is a RNA virus causing respiratory and reproductive diseases in swine. The susceptibility for PRRSV varies between the different breeds of swine. In cell culture, PRRSV virus can be propagated in primary porcine alveolar macrophages and some African green monkey kidney cell lines, such as MARC-145 cells. Previous studies have shown that 3' untranslated region (UTR RNAs of the arteriviruses play an important role in the replication of the virus through interactions with cellular proteins. To better understand the differences in the replication capability of PRRSV in different cell lines, we sought to identify the host cellular proteins interacting with PRRSV 3' UTR RNA. We constructed a cDNA library of MARC-145 cell line in lambda ZAP Express vector and screened the library with the positive sense 3' UTR RNA of PRRSV. Results We found that CD151, a host cellular protein, interacting with PRRSV 3' UTR RNA. The specificity of the interaction between CD151 and PRRSV 3' UTR RNA was examined by gel shift assay as well as North-Western hybridization. The transfection of CD151 expression clone into BHK-21 rendered these cells susceptible to PRRSV infection, and the transfection of siRNA against CD151 into MARC-145 significantly reduced the level of PRRSV infection. Also, anti-CD151 antibody treatment to MARC-145 completely blocked PRRSV infection. Conclusion Based on our results, we suggest that CD151 should cooperate in PRRSV infection in vitro in MARC-145 and BHK-21 cells.

  12. Porcine B-cells recognize epitopes that are conserved between the structural proteins of American- and European-type porcine reproductive and respiratory syndrome virus

    DEFF Research Database (Denmark)

    Oleksiewicz, Martin B.; Bøtner, Anette; Normann, Preben

    2002-01-01

    By selecting phage display libraries with immune sera from experimentally infected pigs, porcine B-cell epitopes in the open reading frame (ORF) 2, 3, 5 and 6 proteins of European-type porcine reproductive and respiratory syndrome virus (PRRSV) were identified. The sequences of all the epitopes...... were well conserved in European-type PRRSV and even between European- and American-type PRRSV. Accordingly, sera from pigs infected with American-type PRRSV cross-reacted with the European-type epitopes. Thus, this study showed, for the first time, the presence of highly conserved epitopes...... epitopes was subjected to closer scrutiny. A heptad motif, VSRRIYQ, which is present in a single copy in ORF2 and 3 proteins, was identified; this arrangement is completely conserved in all European-type PRRSV sequences available. The VSRRIYQ repeat motif colocalized closely with one of the ORF2 epitopes...

  13. Yersinia enterocolitica of porcine origin: carriage of virulence genes and genotypic diversity.

    Science.gov (United States)

    Tadesse, Daniel A; Bahnson, Peter B; Funk, Julie A; Morrow, W E Morgan; Abley, Melanie J; Ponte, Valeria A; Thakur, Siddhartha; Wittum, Thomas; DeGraves, Fred J; Rajala-Schultz, Paivi J; Gebreyes, Wondwossen A

    2013-01-01

    Yersinia enterocolitica is an important foodborne pathogen, and pigs are recognized as a major reservoir and potential source of pathogenic strains to humans. A total of 172 Y. enterocolitica recovered from conventional and antimicrobial-free pig production systems from different geographic regions (North Carolina, Ohio, Michigan, Wisconsin, and Iowa) were investigated to determine their pathogenic significance to humans. Phenotypic and genotypic diversity of the isolates was assessed using antibiogram, serogrouping, and amplified fragment length polymorphism (AFLP). Carriage of chromosomal and plasmid-borne virulence genes were investigated using polymerase chain reaction. A total of 12 antimicrobial resistance patterns were identified. More than two-thirds (67.4%) of Y. enterocolitica were pan-susceptible, and 27.9% were resistant against β-lactams. The most predominant serogroup was O:3 (43%), followed by O:5 (25.6%) and O:9 (4.1%). Twenty-two of 172 (12.8%) isolates were found to carry Yersinia adhesion A (yadA), a virulence gene encoded on the Yersinia virulence plasmid. Sixty-nine (40.1%) isolates were found to carry ail gene. The ystA and ystB genes were detected in 77% and 26.2% of the strains, respectively. AFLP genotyping of isolates showed wide genotypic diversity and were grouped into nine clades with an overall genotypic similarity of 66.8-99.3%. AFLP analysis revealed that isolates from the same production system showed clonal relatedness, while more than one genotype of Y. enterocolitica circulates within a farm.

  14. Cytokine profiles in pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome

    Science.gov (United States)

    In spite of extensive research, immunologic control mechanisms against Porcine Reproductive and Respiratory Syndrome virus (PRRSv) remain poorly understood. Cytokine responses have been exhaustively studied in nursery pigs and show contradictory results. Since no detailed reports on cytokine respons...

  15. Characterization of polyclonal antibodies against nonstructural protein 9 from the porcine reproductive and respiratory syndrome virus

    Directory of Open Access Journals (Sweden)

    Mengmeng ZHAO,Juanjuan QIAN,Jiexiong XIE,Tiantian CUI,Songling FENG,Guoqiang WANG,Ruining WANG,Guihong ZHANG

    2016-06-01

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS is considered to be one of the most important infectious diseases impacting the swine industry and is characterized by reproductive failure in late term gestation in sows and respiratory disease in pigs of all ages. The nonstructural protein 9 gene, Nsp9, encoding the RNA-dependent RNA polymerase, is generally regarded as fairly conserved when compared to other viral proteins. Antibodies against Nsp9 will be of great importance for the diagnosis and treatment of the causal agent, PRRS virus. A study was undertaken to generate polyclonal antibodies against the immunodominant Nsp9. For this purpose, the Nsp9 was expressed in Escherichia coli and subsequently used as an antigen to immunize New Zealand rabbits. Antiserum was identified via an indirect ELISA, and then verified based on the ability to react with both naturally and artificially expressed Nsp9. Results of virus neutralization test showed that this antiserum could not neutralize the PRRSV. Nevertheless, this antiserum as a diagnostic core reagent should prove invaluable for further investigations into the mechanism of PRRS pathogenesis.

  16. The amino acid residues at 102 and 104 in GP5 of porcine reproductive and respiratory syndrome virus regulate viral neutralization susceptibility to the porcine serum neutralizing antibody.

    Science.gov (United States)

    Fan, Baochao; Liu, Xing; Bai, Juan; Zhang, Tingjie; Zhang, Qiaoya; Jiang, Ping

    2015-06-02

    Porcine reproductive and respiratory syndrome virus (PRRSV) is mainly responsible for the heavy economic losses in pig industry in the world. A number of neutralizing epitopes have been identified in the viral structural proteins GP3, GP4, GP5 and M. In this study, the important amino acid (aa) residues of HP-PRRSV strain BB affecting neutralization susceptibility of antibody were examined using resistant strains generated under neutralizing antibody (NAb) pressure in MARC-145 cells, reverse genetic technique and virus neutralization assay. HP-PRRSV strain BB was passaged under the pressure of porcine NAb serum in vitro. A resistant strain BB34s with 102 and 104 aa substitutions in GP5, which have been predicted to be the positive sites for pressure selection (Delisle et al., 2012), was cloned and identified. To determine the effect of the two aa residues on neutralization, eight recombinant PRRSV strains were generated, and neutralization assay results confirmed that the aa residues 102 and 104 in GP5 played an important role in NAbs against HP-PRRSV in MARC-145 cells and porcine alveolar macrophages. Alignment of GP5 sequences revealed that the variant aa residues at 102 and 104 were frequent among type 2 PRRSV strains. It may be helpful for understanding the mechanism regulating the neutralization susceptibility of PRRSV to the NAbs and monitoring the antigen variant strains in the field. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Strain predominance following exposure of vaccinated and naive pregnant gilts to multiple strains of porcine reproductive and respiratory syndrome virus

    OpenAIRE

    Lager, Kelly M.; Mengeling, William L.; Wesley, Ronald D.

    2003-01-01

    Two studies were performed in order to test the relative ability of different strains of porcine reproductive and respiratory syndrome virus (PRRSV) to replicate and cross the placental barrier in pregnant gilts. Study 1 comprised 6 nonvaccinated gilts. Study 2 comprised 8 nonvaccinated gilts and 12 gilts that were vaccinated twice before conception. On, or about, gestation day 90 all gilts were simultaneously exposed to 20 field strains of PRRSV (all strains were distinguishable by restricti...

  18. Virological and immunological responses to porcine reproductive and respiratory syndrome virus in a large population of gilts

    OpenAIRE

    Batista, Laura; Pijoan, Carlos; Dee, Scott; Olin, Michael; Molitor, Thomas; Joo, Han Soo; Xiao, Zhenguo; Murtaugh, Michael

    2004-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes a prolonged active infection followed by a persistent infection in lymphoid tissues lasting for several months. Pigs develop both an antibody and cell-mediated immune response following PRRSV infection, but the specific role of each type in the development of protective immunity and clearance of the virus is not yet known. The aims of this study were to characterize the dynamics of PRRSV persistence from 0 to 135 d post infect...

  19. Transcriptome Differences in Porcine Alveolar Macrophages from Tongcheng and Large White Pigs in Response to Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus (PRRSV Infection

    Directory of Open Access Journals (Sweden)

    Wan Liang

    2017-07-01

    Full Text Available Porcine reproductive and respiratory syndrome virus (PRRSV is a single-stranded positive-sense RNA virus that can cause devastating reproductive failure and respiratory tract lesions, which has led to serious damage to the swine industry worldwide. Our previous studies have indicated that Tongcheng (TC pigs, a Chinese local breed, have stronger resistance or tolerance to PRRSV infection than Large White (LW pigs. This study aims to investigate their host transcriptome differences in porcine alveolar macrophages (PAMs at 7 days post challenge. Transcriptome profiling of PAMs from PRRSV infected and control pigs of these two breeds were performed using RNA-sequencing. For both breeds, there were 1257 common differentially expressed genes (DEGs in response to PRRSV infection, involving hepatic fibrosis/hepatic stellate cell activation, phospholipase C, and granulocyte adhesion and diapedesis pathways. For TC pig, 549 specific DEGs were identified, including VAV2, BCL2 and BAX, which were enriched in activation of leukocyte extravasation and suppression of apoptosis. While, 898 specific DEGs were identified in LW pigs, including GNAQ, GNB5, GNG2, CALM4 and RHOQ, which were involved in suppression of Gαq and PI3K-AKT signaling. This study provides an insight into the transcriptomic comparison of resistant and susceptible pigs to PRRSV infection. TC pigs may promote the extravasation and migration of leukocytes to defend against PRRSV infections and suppress apoptosis of the infected macrophages to increase antigen presentation, thereby reducing the lung lesions.

  20. Immune responses in piglets infected with highly pathogenic porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Wang, Gang; Song, Tengfei; Yu, Ying; Liu, Yonggang; Shi, Wenda; Wang, Shujie; Rong, Fulong; Dong, Jianguo; Liu, He; Cai, Xuehui; Zhou, En-Min

    2011-08-15

    Porcine reproductive and respiratory syndrome virus (PRRSV) infection compromises the host's innate and adaptive immunity. The aim of this study was to investigate the immune responses of piglets infected with highly pathogenic (HP) PRRSV (HuN4 strain) with or without the immunization with CH-1R attenuated PRRSV vaccine. The response was evaluated for the clinical signs, pathological changes and virus load in immune organs, antibody responses and levels of serum IFN-γ, IL-4 and IL-10. The result showed that in comparison with the piglets received the immunization, the piglets infected with HP-PRRSV alone had the thymus atrophy, decreased serum levels of IL-4 and increased serum levels of IL-10 and INF-γ. These results suggest that elevated IL-10 levels at the early stage of the infection may enhance virus survival and delay the induction of protective immunity, while increased levels of IL-4 induce the effective immune responses and increase the animals' health status. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. Economic analysis of outbreaks of porcine reproductive and respiratory syndrome virus in nine sow herds.

    Science.gov (United States)

    Nieuwenhuis, N; Duinhof, T F; van Nes, A

    2012-03-03

    The economic losses due to porcine reproductive and respiratory syndrome virus (PRRSv) outbreaks are reported in the literature to be substantially high, but recent figures are not available. The aim of this study was to quantify the economic effects of epidemic PRRSv outbreaks in Dutch sow herds. Nine sow herds were selected based on a confirmed PRRSv outbreak within those populations. The economic impact during the first 18 weeks after the outbreak was estimated by comparing the overall costs between pre- and postoutbreak periods, using different factors (production data, medication, diagnostics, labour, etc.). An outbreak of PRRSv resulted in a reduced number of sold pigs per sow of 1.7. The economic loss varied between €59 and €379 for one sow per 18-week period outbreak. The mean loss per sow per outbreak was €126. The costs after the outbreak varied significantly from €3 to 160 per sow, due to the different methods used by farmers to tackle PRRSv outbreaks. The calculated costs in this study correlate with the costs of the initial outbreak in The Netherlands of 98 per sow.

  2. Oral immunogenicity of porcine reproductive and respiratory syndrome virus antigen expressed in transgenic banana.

    Science.gov (United States)

    Chan, Hui-Ting; Chia, Min-Yuan; Pang, Victor Fei; Jeng, Chian-Ren; Do, Yi-Yin; Huang, Pung-Ling

    2013-04-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a persistent threat of economically significant influence to the swine industry worldwide. Recombinant DNA technology coupled with tissue culture technology is a viable alternative for the inexpensive production of heterologous proteins in planta. Embryogenic cells of banana cv. 'Pei chiao' (AAA) have been transformed with the ORF5 gene of PRRSV envelope glycoprotein (GP5) using Agrobacterium-mediated transformation and have been confirmed. Recombinant GP5 protein levels in the transgenic banana leaves were detected and ranged from 0.021%-0.037% of total soluble protein. Pigs were immunized with recombinant GP5 protein by orally feeding transgenic banana leaves for three consecutive doses at a 2-week interval and challenged with PRRSV at 7 weeks postinitial immunization. A vaccination-dependent gradational increase in the elicitation of serum and saliva anti-PRRSV IgG and IgA was observed. Furthermore, significantly lower viraemia and tissue viral load were recorded when compared with the pigs fed with untransformed banana leaves. The results suggest that transgenic banana leaves expressing recombinant GP5 protein can be an effective strategy for oral delivery of recombinant subunit vaccines in pigs and can open new avenues for the production of vaccines against PRRSV. © 2012 The Authors Plant Biotechnology Journal © 2012 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

  3. Survival of porcine reproductive and respiratory syndrome virus in fresh pork.

    Science.gov (United States)

    Guarino, Helena; Moura, Junior; Cox, Ryan B; Goyal, Sagar M; Patnayak, Devi P

    2014-06-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) remains one of the most economically important diseases of pigs. Transmission of PRRS virus has been reported through many routes, with aerosol route being the most predominant. There may also be a potential risk of transmission through contami-nated pork, but this has never been investigated. The purpose of this study was to experimentally contaminate fresh pork with three different concentrations of PRRSV and to study virus survival at ambient (25 °C), refrigerated (4 °C), and frozen (-20 °C) temperatures. Concentrations of virus representing natural infectivity level and 'worst case scenario' were studied. The virus was detected in fresh pork at all three virus concentrations for up to 48 h at ambient temperature. At 4 °C, the virus survived for 6 days in pork inoculated with the higher virus concentration and for 3 days in pork inoculated at the lower concentration. At frozen temperature, PRRSV was detected for up to 60 days in pork inoculated at the higher concentration and for 7 days in pork inoculated at the lower concentration. These results suggest that fresh pork has the potential to be a vehicle for virus dissemination depending upon temperature and time of storage.

  4. Homologous challenge of porcine reproductive and respiratory syndrome virus immunity in pregnant swine.

    Science.gov (United States)

    Lager, K M; Mengeling, W L; Brockmeier, S L

    1997-11-01

    The clinical consequences of single or multiple exposure of pregnant gilts to porcine reproductive and respiratory syndrome virus (PRRSV) at various stages of gestation were determined. Thirty-three pregnant gilts were allotted to 6 experimental groups (5 to 7 gilts/group). Gilts of groups 1 to 5 were exposed to strain NADC-8 of PRRSV at the following times: group 1, gestation day (GD) 1; group 2, GDs 1 and 90; group 3, GD 30; group 4, GDs 30 and 90; group 5, GD 90. Virus exposure was by either intrauterine (GD 1) or oronasal (GDs 30 and 90) inoculation. Gilts of group 6 were kept as nonexposed controls. Gilts were either necropsied on or about GD 111 (groups 1 to 5) or were allowed to farrow (group 6). The detection of PRRSV in serum of fetuses and piglets (within 12 hof birth) was considered evidence of transplacental infection. Transplacental infection and virus-induced death were and were not confirmed for groups 3, 4, and 5 and for groups 1, 2, and 6, respectively. Collectively, the results indicated that intrauterine exposure to PRRSV at GD 1 was without clinical effect (groups 1 and 2) and provided protection against subsequent exposure to the same strain of virus at GD 90 (group 2). The highest incidence of transplacental infection and fetal death followed a single exposure to PRRSV at GD 90 (group 5).

  5. Mapping codon usage of the translation initiation region in porcine reproductive and respiratory syndrome virus genome

    Directory of Open Access Journals (Sweden)

    Dou Yong-xi

    2011-10-01

    Full Text Available Abstract Background Porcine reproductive and respitatory syndrome virus (PRRSV is a recently emerged pathogen and severely affects swine populations worldwide. The replication of PRRSV is tightly controlled by viral gene expression and the codon usage of translation initiation region within each gene could potentially regulate the translation rate. Therefore, a better understanding of the codon usage pattern of the initiation translation region would shed light on the regulation of PRRSV gene expression. Results In this study, the codon usage in the translation initiation region and in the whole coding sequence was compared in PRRSV ORF1a and ORFs2-7. To investigate the potential role of codon usage in affecting the translation initiation rate, we established a codon usage model for PRRSV translation initiation region. We observed that some non-preferential codons are preferentially used in the translation initiation region in particular ORFs. Although some positions vary with codons, they intend to use codons with negative CUB. Furthermore, our model of codon usage showed that the conserved pattern of CUB is not directly consensus with the conserved sequence, but shaped under the translation selection. Conclusions The non-variation pattern with negative CUB in the PRRSV translation initiation region scanned by ribosomes is considered the rate-limiting step in the translation process.

  6. Immunological Features of the Non-Structural Proteins of Porcine Reproductive and Respiratory Syndrome Virus

    Directory of Open Access Journals (Sweden)

    Edgar Rascón-Castelo

    2015-02-01

    Full Text Available Porcine reproductive and respiratory syndrome virus (PRRSV is currently one of the most important viruses affecting the swine industry worldwide. Despite the large number of papers published each year, the participation of non-structural proteins (nsps in the immune response is not completely clear. nsps have been involved in the host innate immune response, specifically, nsp1α/β, nsp2, nsp4 and nsp11 have been associated with the immunomodulation capability of the virus. To date, only participation by nsp1, nsp2, nsp4 and nsp7 in the humoral immune response has been reported, with the role of other nsps being overlooked. Furthermore, nsp1, nsp2, nsp5, nsp7 nsp9, nsp10, nsp11 have been implicated in the induction of IFN-γ and probably in the development of the cell-mediated immune response. This review discusses recent reports involving the participation of nsps in the modulation of the innate immune response and their role in the induction of both the humoral and cellular immune responses.

  7. Porcine reproductive and respiratory syndrome virus induces apoptosis through a mitochondria-mediated pathway.

    Science.gov (United States)

    Lee, Sang-Myeong; Kleiboeker, Steven B

    2007-09-01

    As with a number of other viruses, Porcine reproductive and respiratory syndrome virus (PRRSV) has been shown to induce apoptosis, although the mechanism(s) involved remain unknown. In this study we have characterized the apoptotic pathways activated by PRRSV infection. PRRSV-infected cells showed evidence of apoptosis including phosphatidylserine exposure, chromatin condensation, DNA fragmentation, caspase activation (including caspase-8, 9, 3), and PARP cleavage. DNA fragmentation was dependent on caspase activation but blocking apoptosis by a caspase inhibitor did not affect PRRSV replication. Upregulation of Bax expression by PRRSV infection was followed by disruption of the mitochondria transmembrane potential, resulting in cytochrome c redistridution to the cytoplasm and subsequent caspase-9 activation. A crosstalk between the extrinsic and intrinsic pathways was demonstrated by dependency of caspase-9 activation on active caspase-8 and by Bid cleavage. Furthermore, in this study we provide evidence of the possible involvement of reactive oxygen species (ROS)-mediated oxidative stress in apoptosis induced by PRRSV. Our data indicated that cell death caused by PRRSV infection involves necrosis as well as apoptosis. In summary, these findings demonstrate mechanisms by which PRRSV induces apoptosis and will contribute to an enhanced understanding of PRRSV pathogenesis.

  8. Synergism between porcine reproductive and respiratory syndrome virus (PRRSV) and Salmonella choleraesuis in swine.

    Science.gov (United States)

    Wills, R W; Gray, J T; Fedorka-Cray, P J; Yoon, K J; Ladely, S; Zimmerman, J J

    2000-02-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) and Salmonella choleraesuis are two leading causes of economic loss in the swine industry. While respiratory disease is common in both S. choleraesuis and PRRSV infections, the factors that contribute to its development remain largely undefined. We investigated the interaction of PRRSV, S. choleraesuis, and stress in 5-week-old swine. All combinations of three factors (inoculation with S. choleraesuis on Day 0, PRRSV on Day 3, and treatment with dexamethasone on Days 3-7) were used to produce eight treatment groups in two independent trials. Fecal samples, tonsil and nasal swabs, serum samples and postmortem tissues were collected for bacteriologic and virologic examinations. No clinical signs were observed in pigs inoculated with only PRRSV or only S. choleraesuis. In contrast, pigs which were dually infected with S. choleraesuis and PRRSV exhibited unthriftiness, rough hair coats, dyspnea, and diarrhea. The pigs which received all three treatment factors were the most severely affected and 43% (three of seven) of the animals in this group died. Individuals in this group shed significantly higher quantities of S. choleraesuis in feces and had significantly higher serum PRRSV titers compared to other treatments (p syndrome similar to that observed in the field.

  9. INVESTIGATION OF IMMUNE PATHWAYS INVOLVED IN REGULATING RESPONSES TO PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME USING SWINE PROTEIN-ANNOTATED OLIGONUCLEOTIDE MICROARRAYS

    Science.gov (United States)

    Porcine reproductive and respiratory syndrome (PRRS) costs U.S. swine producers around $560 million annually. PRRS virus (PRRSV)-infected pigs are susceptible to pneumonia and reproductive losses; infected sows have increased rates of abortions, stillbirths, mummifications, and give birth to weak pi...

  10. Expression of NK1 receptor at the protein and mRNA level in the porcine female reproductive system.

    Science.gov (United States)

    Bukowski, R

    2014-01-01

    The presence and distribution of substance P (SP) receptor NK1 was studied in the ovary, the oviduct and the uterus (uterine horn and cervix) of the domestic pig using the methods of molecular biology (RT-PCR and immunoblot) and immunohistochemistry. The expression of NK1 receptor at mRNA level was confirmed with RT-PCR in all the studied parts of the porcine female reproductive system by the presence of 525 bp PCR product and at the protein level by the detection of 46 kDa protein band in immunoblot. Immunohistochemical staining revealed the cellular distribution of NK1 receptor protein. In the ovary NKI receptor was present in the wall of arterial blood vessels, as well as in ovarian follicles of different stages of development. In the tubular organs the NK1 receptor immunohistochemical stainings were observed in the wall of the arterial blood vessels, in the muscular membrane, as well as in the mucosal epithelium. The study confirmed the presence of NK1 receptor in the tissues of the porcine female reproductive tract which clearly points to the possibility that SP can influence porcine ovary, oviduct and uterus.

  11. Mycoplasma hyorhinis is a potential pathogen of porcine respiratory disease complex that aggravates pneumonia caused by porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Lee, Jung-Ah; Oh, Yu-Ri; Hwang, Min-A; Lee, Joong-Bok; Park, Seung-Yong; Song, Chang-Seon; Choi, In-Soo; Lee, Sang-Won

    2016-09-01

    The porcine respiratory disease complex (PRDC) caused by numerous bacterial and viral agents has a great impact on pig industry worldwide. Although Mycoplasma hyorhinis (Mhr) has been frequently isolated from lung lesions from pigs with PRDC, the pathological importance of Mhr may have been underestimated. In this study, 383 serum samples obtained from seven herds with a history of PRDC were tested for specific antibodies to Mhr, Mycoplasma hyopneumoniae (Mhp), and porcine reproductive and respiratory syndrome virus (PRRSV). Seropositive rates of PRRSV were significantly correlated with those of Mhr (correlation coefficient, 0.862; P-value, 0.013), but not with those of Mhp (correlation coefficient, -0.555; P-value, 0.196). In vivo experiments demonstrated that pigs co-infected with Mhr and PRRSV induced more severe lung lesions than pigs infected with Mhr or PRRSV alone. These findings suggest that Mhr is closely associated with pneumonia caused by PRRSV and provide important information on Mhr pathogenesis within PRDC. Therefore, effective PRDC control strategies should also consider the potential impact of Mhr in the pathogenesis of PRDC. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Investigating the financial impact of porcine reproductive and respiratory syndrome on smallholder pig farmers in Cambodia.

    Science.gov (United States)

    Zhang, A; Young, J R; Suon, S; Ashley, K; Windsor, P A; Bush, R D

    2017-04-01

    Porcine reproductive and respiratory syndrome (PRRS) was first identified in Cambodia in 2010, causing serious problems on affected farms, although the costs of the disease have not been well defined. The household financial impact of a PRRS outbreak in Cambodia was investigated using partial budget analysis, examining the economic benefit of three proposed interventions: (i) quarterly PRRS vaccine use, (ii) biosecurity implementation, and (iii) implementation of vaccination and biosecurity. The analyses were applied to three farm models: (i) a two-sow breeder; (ii) a five-pig fattener; and (iii) a single-sow, three-pig farrow-to-finish/breeder. Data was derived from a knowledge, attitude, and practice survey of 240 smallholder farmers (61 with pigs) from 16 villages across 5 provinces, plus case studies of 12 farmers selected for more detailed financial analysis. The study indicated that financial losses associated with PRRS were severe, with a 25% mean loss to the annual household income of 61 interviewed farmers. Partial budget analysis identified a strongly positive incentive for vaccination and biosecurity to be implemented in combination, with the highest annual net benefit of USD 357.10 realised by the breeder system. However, due to current scarcity of the PRRS vaccine and its high cost to smallholders, biosecurity interventions may be more cost-effective, especially for low PRRS incidence regions. It was concluded that PRRS critically constrains the profitability of smallholder pig farms and that these findings will assist development of village-level livestock disease risk management programmes that encourage adoption of vaccination and biosecurity practices to enhance farmer livelihoods in Cambodia.

  13. Chinese highly pathogenic porcine reproductive and respiratory syndrome virus exhibits more extensive tissue tropism for pigs

    Directory of Open Access Journals (Sweden)

    Li Limin

    2012-09-01

    Full Text Available Abstract Background The highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV emerging in China exhibits high fatality to pigs. However, the mechanism related to the increased pathogenicity of the virus remains unclear. In the present study, the differences in tissue tropism between the highly pathogenic PRRSV strain (JXwn06 and the low pathogenic PRRSV strain (HB-1/3.9 were investigated using PRRSV-specific immunohistochemistry (IHC staining to provide evidence for elucidating possible mechanism of the pathogenicity of Chinese highly pathogenic PRRSV. Findings IHC examination showed that PRRSV antigen in the tissues including spleen, tonsil, thymus, kidney, cerebellum, stomach, small intestine, large intestine, turbinal bone and laryngeal cartilage was positive in more pigs inoculated with JXwn06 than HB-1/3.9, and the tissues including trachea, esophagus, liver, mandibular gland and thyroid gland were positive for viral antigen in the pigs inoculated with JXwn06, but not in the pigs inoculated with HB-1/3.9. Meanwhile, we observed that epithelium in tissues including interlobular bile duct in liver, distal renal tubule of kidney, esophageal gland and tracheal gland were positive for viral antigen only in JXwn06-inoculated pigs, and epithelium of gastric mucosa and fundic gland, and intestinal gland were positive for viral antigen in both JXwn06- and HB-1/3.9-inoculated pigs, using monoclonal antibodies to N and Nsp2 proteins. Conclusions Taken together, these findings indicate that the highly pathogenic PRRSV JXwn06 displays an expanded tissue tropism in vivo, suggesting this may contribute to its high pathogenicity to pigs.

  14. Control of porcine reproductive and respiratory syndrome (PRRS through genetic improvements in disease resistance and tolerance

    Directory of Open Access Journals (Sweden)

    Raymond eRowland

    2012-12-01

    Full Text Available Infections caused by porcine reproductive and respiratory syndrome virus (PRRSV have a severe economic impact on pig production in North America, Europe and Asia. The emergence and eventual predominance of PRRS in the 1990s are the likely result of changes in the pork industry initiated in the late 1970s, which allowed the virus to occupy a unique niche within a modern commercial production system. PRRSV infection is responsible for severe clinical disease, but can maintain a life-long subclinical infection, as well as participate in several polymicrobial syndromes. Current vaccines lessen clinical signs, but are of limited use for disease control and elimination. The relatively poor protective immunity following vaccination is a function of the virus’s capacity to generate a large degree of genetic diversity, combined with several strategies to evade innate and adaptive immune responses. In 2007, the PRRS Host Genetics consortium (PHGC was established to explore the role of host genetics as as an avenue for PRRS control. The PHGC model for PRRS incorporates the experimental infection of large numbers of growing pigs and has created the opportunity to study experimental PRRSV infection at the population level. The results show that pigs can be placed into distinct phenotypic groups, including pigs that show resistance or pigs that exhibit tolerance to infection. Tolerance is best illustrated by pigs that gain weight normally in the face of a relatively high virus load. Genome-wide association analysis has identified a region on chromosome 4 (SSC4 correlated with resistance; i.e., higher weight gain combined with lower virus load. The genomic region is near a family of genes involved in innate immunity. These results create the opportunity to develop breeding programs that will produce pigs with increased resistance to PRRS. The identification of genomic markers involved in tolerance will likely prove more difficult, primarily because tolerance

  15. Duration of homologous porcine reproductive and respiratory syndrome virus immunity in pregnant swine.

    Science.gov (United States)

    Lager, K M; Mengeling, W L; Brockmeier, S L

    1997-11-01

    The duration of porcine reproductive and respiratory syndrome virus (PRRSV) homologous immunity was tested in this study and found to last for at least 604 days post experimental exposure to field PRRSV. Eleven gilts (group A) received a primary exposure to field PRRSV by either an oronasal (n = 6) or an intrauterine (n = 5) route. The gilts were naturally bred at selected times (143 to 514 days) after primary virus exposure. They were oronasally exposed a second time to the same strain of virus on or about gestation day 90. Ten age-matched control sows free of PRRSV-specific antibody from the same source farm (group B) were naturally bred and were oronasally exposed to aliquots of the homologous challenge virus on or about gestation day 90. Nine of the 11 gilts in group A and all animals in group B became pregnant following one breeding cycle. The two nonpregnant gilts in group A were each naturally bred during four additional estrus cycles and neither became pregnant. They were exposed to homologous challenge virus 562 and 604 days post primary exposure, respectively. All animals were necropsied 21 days post homologous challenge. Sera and alveolar macrophages from each dam, and sera from each fetus were tested for virus. Transplacental infection was detected in 0/9 and 8/10 litters in groups A and B, respectively. Virus was detected in 0/11 and 10/10 of the alveolar macrophage samples collected in groups A and B, respectively. Serum was harvested at selected times throughout the experiment and tested for PRRSV-specific antibody by indirect immunofluorescence microscopy. All gilts in group A were seropositive for the duration of the experiment, and all animals in group B seroconverted following exposure to field PRRSV. This study shows that adult swine can produce a homologous protective immunity after PRRSV exposure that may persist for the production life of the animal.

  16. Antigenic structure of the nucleocapsid protein of porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Wootton, S K; Nelson, E A; Yoo, D

    1998-11-01

    A collection of 12 monoclonal antibodies (MAbs) raised against porcine reproductive and respiratory syndrome (PRRS) virus was used to study the antigenic structure of the virus nucleocapsid protein (N). The full-length N gene, encoded by open reading frame 7, was cloned from the Canadian PRRS virus, PA-8. Deletions were introduced into the N gene to produce a series of nine overlapping protein fragments ranging in length from 25 to 112 amino acids. The individual truncated genes were cloned as glutathione S-transferase fusions into a eukaryotic expression vector downstream of the T7 RNA polymerase promoter. HeLa cells infected with recombinant vaccinia virus expressing T7 RNA polymerase were transfected with plasmid DNA encoding the N protein fragments, and the antigenicity of the synthesized proteins was analyzed by immunoprecipitation. Based on the immunoreactivities of the N protein deletion mutants with the panel of N-specific MAbs, five domains of antigenic importance were identified. MAbs SDOW17, SR30, and 5H2.3B12.1C9 each identified independent domains defined by amino acids 30 to 52, 69 to 123, and 37 to 52, respectively. Seven of the MAbs tested specifically recognized the local protein conformation formed in part by the amino acid residues 52 to 69. Furthermore, deletion of 11 amino acids from the carboxy terminus of the nucleocapsid protein disrupted the epitope configuration recognized by all of the conformation-dependent MAbs, suggesting that the carboxy-terminal region plays an important role in maintaining local protein conformation.

  17. Sero-Prevalence of Porcine Reproductive and Respiratory Syndrome (PRRS) in Pigs of Different Developmental Regions of Nepal

    OpenAIRE

    Mahesh K.C.; Bhoj Raj Joshi; Swoyam Prakash Shrestha; Meera Prajapati; Dipak Kathayat; Santosh Dhakal

    2015-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is a highly infectious viral disease of swine. This disease is becoming threat to pig industry of every corner of the world. As the status of this disease in Nepal is not properly understood so far, a cross-sectional sero-prevalence study was carried out in four development regions. A total of 200 blood samples were collected from major pig producing areas of eight districts of four development regions of Nepal and the serum samples were te...

  18. RNA recombination in Porcine Reproductive and Respiratory Syndrome Virus is restricted to parental sequences with high similarity

    DEFF Research Database (Denmark)

    Vugt, J.J.F.A. van; Storgaard, T.; Oleksiewicz, M. B.

    2001-01-01

    Two types of porcine reproductive and respiratory syndrome virus (PRRSV) exist, a North American type and a European type. The co-existence of both types in some countries, such as Denmark, Slovakia and Canada, creates a risk of inter-type recombination. To evaluate this risk, cell cultures were co......, but no recombination was detected between the European and North American types. Calculation of the maximum theoretical risk of European–American recombination, based on the sensitivity of the RT–PCR system, revealed that RNA recombination between the European and North American types of PRRSV is at least 10000 times...

  19. High frequency RNA recombination in porcine reproductive and respiratory syndrome virus occurs preferentially between parental sequences with high similarity

    DEFF Research Database (Denmark)

    van Vugt, Joke .J.F.A.; Storgaard, Torben; Oleksiewicz, Martin B.

    2001-01-01

    Two types of porcine reproductive and respiratory syndrome virus (PRRSV) exist, a North American type and a European type. The co-existence of both types in some countries, such as Denmark, Slovakia and Canada, creates a risk of inter-type recombination. To evaluate this risk, cell cultures were co......, but no recombination was detected between the European and North American types. Calculation of the maximum theoretical risk of European-American recombination, based on the sensitivity of the RT-PCR system, revealed that RNA recombination between the European and North American types of PRRSV is at least 10000 times...

  20. Epizootiological and diagnostic significance of porcine reproductive and respiratory syndrome control

    Directory of Open Access Journals (Sweden)

    Radojičić Biljana

    2002-01-01

    Full Text Available The porcine reproductive and respiratory syndrome (PRRS is a new viral disease in swine, designated exclusively under the acronym PRRS by the European Commission in 1991. The cause of this disease was isolated and determined in 1991 at the Lelystad Institute in The Netherlands as Lelystad aretrivirus. The PRRSV is an RNA virus of the order Nidovirales, the family Arteriviridae, the genus Arterivirus (Cavanaugh, 1997. Different genomic and pheriotypic varieties of the virus are significant. It is replicated in macrophages, it induces permanent viraemia, causes the creation of antibodies, and leads to persistent and latent infections. It is isolated from tonsil tissue, alveolar macrophages, the uterus, and fetal homogenate composed of different tissues (Wills et al., 1997. All production categories of swine can contract PRRS, but pregnant sows, suckling piglets and fattening swine are considered endangered categories. Morbidity and mortapty is between 8-80%, which also depends on the animal category. Economic damages are substantial when one considers the high percentage of still-born piglets, mummified fetuses and suckling piglets. Irregular successive cycles in sows are also expressed. In fattening swine, in addition to a respiratory form of the clinical picture, the time period until animals reach abattoir weight is extended even up to 30 days, which is also a considerable economic loss. Costs of treating possible secondary bacterial infections, diagnostics and immunoprophylaxis are not negligible. The OIE placed PRRS on the B list in 1992 as a contagious disease of swine which incurs economic losses in almost all countries of the world. Diagnosis is made by isolating and determining the virus and/or by serodiagnostics (ELISA and PCR. Certain countries have already made up protocols for the implementation of constant diagnostics and suggested eradication measures (Dee S.A. et al., 2000. In our country, the first clinical cases of PRRS were

  1. A Synthetic Porcine Reproductive and Respiratory Syndrome Virus Strain Confers Unprecedented Levels of Heterologous Protection.

    Science.gov (United States)

    Vu, Hiep L X; Ma, Fangrui; Laegreid, William W; Pattnaik, Asit K; Steffen, David; Doster, Alan R; Osorio, Fernando A

    2015-12-01

    Current vaccines do not provide sufficient levels of protection against divergent porcine reproductive and respiratory syndrome virus (PRRSV) strains circulating in the field, mainly due to the substantial variation of the viral genome. We describe here a novel approach to generate a PRRSV vaccine candidate that could confer unprecedented levels of heterologous protection against divergent PRRSV isolates. By using a set of 59 nonredundant, full-genome sequences of type 2 PRRSVs, a consensus genome (designated PRRSV-CON) was generated by aligning these 59 PRRSV full-genome sequences, followed by selecting the most common nucleotide found at each position of the alignment. Next, the synthetic PRRSV-CON strain was generated through the use of reverse genetics. PRRSV-CON replicates as efficiently as our prototype PRRSV strain FL12, both in vitro and in vivo. Importantly, when inoculated into pigs, PRRSV-CON confers significantly broader levels of heterologous protection than does wild-type PRRSV. Collectively, our data demonstrate that PRRSV-CON can serve as an excellent candidate for the development of a broadly protective PRRSV vaccine. The extraordinary genetic variation of RNA viruses poses a monumental challenge for the development of broadly protective vaccines against these viruses. To minimize the genetic dissimilarity between vaccine immunogens and contemporary circulating viruses, computational strategies have been developed for the generation of artificial immunogen sequences (so-called "centralized" sequences) that have equal genetic distances to the circulating viruses. Thus far, the generation of centralized vaccine immunogens has been carried out at the level of individual viral proteins. We expand this concept to PRRSV, a highly variable RNA virus, by creating a synthetic PRRSV strain based on a centralized PRRSV genome sequence. This study provides the first example of centralizing the whole genome of an RNA virus to improve vaccine coverage. This

  2. Genetic analysis of ORF5 porcine reproductive and respiratory syndrome virus isolated in Vietnam.

    Science.gov (United States)

    Thuy, Nguyen Thi Dieu; Thu, Nguyen Thi Dieu; Son, Nguyen Giang; Ha, Le Thi Thu; Hung, Vo Khanh; Nguyen, Nguyen Thao; Khoa, Do Vo Anh

    2013-07-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important swine pathogens because it is highly infectious and causes economic losses due to decreased pig productivity. In this study, the 603 bp complete major envelope protein encoding gene (ORF5) of 32 field PRRSV isolates from Vietnam collected during 2008-2012 were sequenced and analyzed. Multiple nucleotide (nt) and deduced amino acid (aa) alignments of ORF5 were performed on the 32 isolates: the representative strains (European and North American genotypes), Chinese strains available in GenBank and vaccine strains licensed for use in Vietnam. The results showed 94.8-100.0% nt identity and 94.0-100% aa similarity among the 32 isolates. These isolates shared similarities with the prototype of the North American PRRSV strain (VR-2332; nt 87.8-89.3%, aa 87.5-90.0%), and Lelystat virus, the prototype of the European PRRSV strain (LV; nt 61.1-61.9%, aa 55.1-57.0%). There was greater similarity with QN07 (nt 96.5-98.5%, aa 96.0-99.0%) from the 2007 PRRS outbreak in QuangNam Province, CH-1a (nt 93.2-95.1%, 91.5-93.5%) isolated in China in 1995 and JXA1 (nt 96.5-98.6%, aa 95.0-98.0%), the highly pathogenic strain from China isolated in 2006. The Vietnamese isolates were more similar to JXA1-R (nt 96.5-98.6%, aa 95.0-98.0%), the strain used in Chinese vaccines, than to Ingelvac MLV/BSL-PS (nt 87.2-89.0%, aa 86.0-89.0%). Phylogenetic analysis showed that the 32 isolates were of the North American genotype and classified into sub-lineage 8.7. This sub-lineage contains highly pathogenic Chinese PRRSV strains. This study documents genetic variation in circulating PRRSV strains and could assist more effective use of PRRS vaccines in Vietnam. © 2013 The Societies and Wiley Publishing Asia Pty Ltd.

  3. Generation and immunogenicity of porcine circovirus type 2 chimeric virus-like particles displaying porcine reproductive and respiratory syndrome virus GP5 epitope B.

    Science.gov (United States)

    Hu, Gaowei; Wang, Naidong; Yu, Wanting; Wang, Zhanfeng; Zou, Yawen; Zhang, Yan; Wang, Aibing; Deng, Zhibang; Yang, Yi

    2016-04-07

    Virus-like particles (VLPs) can be used as transfer vehicles carrying foreign proteins or antigen epitopes to produce chimeric VLPs for bivalent or multivalent vaccines. Based on the crystal structure of porcine circovirus type 2 (PCV2) capsid protein (Cap), in addition to alignment of the Cap sequences collected from various isolates of PCV2 and PCV1, we predicted that Loop CD of the PCV2 Cap should tolerate insertion of foreign epitopes, and furthermore that such an insertion could be presented on the surface of PCV2 VLPs. To validate this, the GP5 epitope B of porcine reproductive and respiratory syndrome virus (PRRSV) was inserted into Loop CD of the PCV2 Cap. The 3D structure of the recombinant PCV2 Cap (rCap) was simulated by homology modeling; it appeared that the GP5 epitope B was folded as a relatively independent unit, separated from the PCV2 Cap backbone. Furthermore, based on transmission electron microscopy, the purified PCV2 rCap self-assembled into chimeric VLPs which entered PK-15 cells. In addition, PCV2 chimeric VLPs induced strong humoral (neutralizing antibodies against PCV2 and PRRSV) and cellular immune responses in mice. We concluded that the identified insertion site in the PCV2 Cap had great potential to develop PCV2 VLPs-based bivalent or multivalent vaccines; furthermore, it would also facilitate development of a nano-device to present a functional peptide on the surface of the VLPs that could be used for therapeutic purposes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Experimental inoculation of swine at various stages of gestation with a Danish isolate of porcine reproductive and respiratory syndrome virus (PRRSV)

    DEFF Research Database (Denmark)

    Kranker, Søren; Nielsen, Jens; Bille-Hansen, Vivi

    1998-01-01

    Following intranasal inoculation of three groups of pregnant swine (in total 11 dams) with a Danish isolate of porcine reproductive and respiratory syndrome virus (PRRSV) on or about day 85, 70 and 45 of gestation, respectively, reproductive disturbances were observed in the first two groups...... inflammatory conditions, especially in the lung and heart. In conclusion, the present results support the hypothesis, that PRRSV infection of dams late in pregnancy has the greatest likelihood of transplacental infection of fetuses....

  5. Identification of radically different variants of porcine reproductive and respiratory syndrome virus in Eastern Europe: towards a common ancestor for European and American viruses

    DEFF Research Database (Denmark)

    Stadejek, T.; Stankevicius, A.; Storgaard, Torben

    2002-01-01

    We determined 22 partial porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 sequences, representing pathogenic field strains mainly from Poland and Lithuania, and two currently available European-type live PRRSV vaccines. Also, the complete ORF7 of two Lithuanian and two Polish...

  6. Temporary CD8(+) T-cell depletion in pigs does not exacerbate infection with porcine reproductive and respiratory syndrome virus (PRRSV)

    DEFF Research Database (Denmark)

    Lohse, Louise; Nielsen, Jens; Eriksen, Lis

    2004-01-01

    Several studies have demonstrated a consistent increase in the CD8(+) T-cell subset of pigs following infection with porcine reproductive and respiratory virus (PRRSV). Consequently, it has been suggested that CD8(+) T-cells may play an important role in protection against this infection. In order...

  7. Phenotypic modulation and cytokine profiles of antigen presenting cells by European subtype 1 and 3 porcine reproductive and respiratory syndrome virus strains in vitro and in vivo

    NARCIS (Netherlands)

    Weesendorp, E.; Stockhofe-Zurwieden, N.; Popma-de Graaf, D.J.; Fijten, H.P.D.; Rebel, J.M.J.

    2013-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes continuous problems in the pig industry, due to high costs of outbreaks and reduced welfare of diseased pigs. The severity of infection is, partly, dependent on the virus strain. Recently isolated Eastern-European subtype 3 strains

  8. The Non-structural Protein 5 and Matrix Protein Are Antigenic Targets of T Cell Immunity to Genotype 1 Porcine Reproductive and Respiratory Syndrome Viruses

    DEFF Research Database (Denmark)

    Mokhtar, Helen; Pedrera, Miriam; Frossard, Jean-Pierre

    2016-01-01

    The porcine reproductive and respiratory syndrome virus (PRRSV) is the cause of one of the most economically important diseases affecting swine worldwide. Efforts to develop a next-generation vaccine have largely focused on envelope glycoproteins to target virus-neutralizing antibody responses. H...

  9. A case-control questionnaire survey of risk factors for Porcine Reproductive and Respiratory Syndrome (PRRS) seropositi¬vity in Danish swine herds

    DEFF Research Database (Denmark)

    Mousing, J.; Permin, A.; Mortensen, S.

    1997-01-01

    Sixty-eight case herds seropositive to porcine reproductive and respiratory syndrome (PRRS) were compared to 128 seronegative controls in a double-blinded questionnaire survey. The study indicated no increased risk of PRRS seropositivity for herds using artificial insemination with semen from PRRS...

  10. Genetic and antigenic characterization of complete genomes of Type 1 Porcine Reproductive and Respiratory Syndrome viruses (PRRSV) isolated in Denmark over a period of 10 years

    DEFF Research Database (Denmark)

    Kvisgaard, Lise Kirstine; Hjulsager, Charlotte Kristiane; Kristensen, Charlotte

    2013-01-01

    Porcine Reproductive and Respiratory Syndrome (PRRS) caused by the PRRS virus (PRRSV) is considered one of the most devastating swine diseases worldwide. PRRS viruses are divided into two major genotypes, Type 1 and Type 2, with pronounced diversity between and within the genotypes. In Denmark more...

  11. The major envelope protein, GP(5), of a European porcine reproductive and respiratory syndrome virus contains a neutralization epitope in its N-terminal ectodomain

    NARCIS (Netherlands)

    Wissink, E.H.J.; Wijk, van H.A.R.; Kroese, M.V.; Weiland, E.; Meulenberg, J.J.M.; Rottier, P.J.M.; Rijn, van P.A.

    2003-01-01

    A set of neutralizing monoclonal antibodies (mAbs) directed against the GP5 protein of European type porcine reproductive and respiratory syndrome virus (PRRSV) has been produced previously (Weiland et al., 1999). This set reacted with a plaque-purified virus (PPV) subpopulation of Dutch isolate

  12. Significance of the oligosaccharides of the porcine reproductive and respiratory syndrome virus glycoproteins GP2a and GP5 for infectious virus production

    NARCIS (Netherlands)

    Wissink, E.H.J.; Kroese, M.V.; Maneschijn-Bonsing, J.G.; Meulenberg, J.J.; Rijn, van P.A.; Rijsewijk, F.A.M.; Rottier, P.J.M.

    2004-01-01

    The arterivirus porcine reproductive and respiratory syndrome virus (PRRSV) contains four glycoproteins, GP2a, GP3, GP4 and GP5, the functions of which are still largely unresolved. In this study, the significance of the N-glycosylation of the GP2a and GP5 proteins of PRRSV strain LV was

  13. The nsp1 alpha and nsp1 beta papain-like autoproteinases are essential for porcine reproductive and respiratory syndrome virus RNA synthesis

    NARCIS (Netherlands)

    Kroese, M.V.; Zevenhoven-Dobbe, J.C.; Ruijter, J.N.A.B.D.; Peeters, B.P.H.; Meulenberg, J.J.M.; Cornelissen, A.H.M.; Snijder, E.J.

    2008-01-01

    The two N-terminal cleavage products, nsp1 alpha and nsp1 beta, of the replicase polyproteins of porcine reproductive and respiratory syndrome virus (PRRSV) each contain a papain-like autoproteinase domain, which have been named PCP alpha and PCP beta, respectively. To assess their role in the PRRSV

  14. Changes in leukocyte subsets of pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome

    Science.gov (United States)

    In spite of more than two decades of extensive research, the understanding of porcine reproductive and respiratory syndrome virus (PRRSv) immunity is still incomplete. A PRRSv infection of the late term pregnant female can result in abortions, early farrowings, fetal death, and the birth of weak, co...

  15. Transcriptome profile of lung dendritic cells after in vitro porcine reproductive and respiratory syndrome virus (PRRSV) infection.

    Science.gov (United States)

    Pröll, Maren Julia; Neuhoff, Christiane; Schellander, Karl; Uddin, Muhammad Jasim; Cinar, Mehmet Ulas; Sahadevan, Sudeep; Qu, Xueqi; Islam, Md Aminul; Poirier, Mikhael; Müller, Marcel A; Drosten, Christian; Tesfaye, Dawit; Tholen, Ernst; Große-Brinkhaus, Christine

    2017-01-01

    The porcine reproductive and respiratory syndrome (PRRS) is an infectious disease that leads to high financial and production losses in the global swine industry. The pathogenesis of this disease is dependent on a multitude of factors, and its control remains problematic. The immune system generally defends against infectious diseases, especially dendritic cells (DCs), which play a crucial role in the activation of the immune response after viral infections. However, the understanding of the immune response and the genetic impact on the immune response to PRRS virus (PRRSV) remains incomplete. In light of this, we investigated the regulation of the host immune response to PRRSV in porcine lung DCs using RNA-sequencing (RNA-Seq). Lung DCs from two different pig breeds (Pietrain and Duroc) were collected before (0 hours) and during various periods of infection (3, 6, 9, 12, and 24 hours post infection (hpi)). RNA-Seq analysis revealed a total of 20,396 predicted porcine genes, which included breed-specific differentially expressed immune genes. Pietrain and Duroc infected lung DCs showed opposite gene expression courses during the first time points post infection. Duroc lung DCs reacted more strongly and distinctly than Pietrain lung DCs during these periods (3, 6, 9, 12 hpi). Additionally, cluster analysis revealed time-dependent co-expressed groups of genes that were involved in immune-relevant pathways. Key clusters and pathways were identified, which help to explain the biological and functional background of lung DCs post PRRSV infection and suggest IL-1β1 as an important candidate gene. RNA-Seq was also used to characterize the viral replication of PRRSV for each breed. PRRSV was able to infect and to replicate differently in lung DCs between the two mentioned breeds. These results could be useful in investigations on immunity traits in pig breeding and enhancing the health of pigs.

  16. Five Years Seroprevalence Study of Porcine Reproductive and Respiratory Syndrome Virus in Lithuanian Pig and Wild Boar Populations

    Directory of Open Access Journals (Sweden)

    Stankevičius Arunas

    2014-10-01

    Full Text Available Serological study of porcine reproductive and respiratory syndrome virus (PRRSV infection in pigs and wild boars was conducted in Lithuania between 2009 and 2013. Antibody level was measured using a commercial ELISA. The 4.32% (95% CI 3.92-4.72 out of 9856 examined porcine sera were positive for the PRRSV antibodies. The antibodies were detected in 11.82% (95% CI 10.28-13.36 of all investigated serum samples of sows and gilts. As much as 8.2% of serologically positive samples (95% CI 6.83-9.57 were determined in the piglets under three months of age. Considerably smaller (P < 0.05 seroprevalence was detected in boars (0.62% and fattening pigs (1.84%. From 1357 examined sera of wild boar, collected between 2009 and 2013 hunting seasons, 5.38% (95% CI 4.52-8.2 of samples were positive for PRRSV antibodies in 23 locations out of 50 investigated. The analysis of seroprevalence in different age groups of wild boars showed that PRRSV antibodies were detected in all age groups; however, it was significantly higher in adults than in juveniles or subadults and reached up to 10.02% (95% CI 7.39-12.65.

  17. Blood antioxidant enzymes (SOD, GPX), biochemical and haematological parameters in pigs naturally infected with porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Stukelj, M; Toplak, I; Svete, A Nemec

    2013-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has become one of the most economically important diseases for the swine industry worldwide. The objective of the study was to determine selected blood antioxidant enzymes (glutathione peroxidase (GPX), superoxide dismutase (SOD)), biochemical and haematological parameters in PRRS positive and negative pigs of three different categories, mainly to test oxidative stress hypothesis in pigs naturally infected with PRRS virus. Ninety PRRS positive and 90 PRRS negative pigs were included in the study. The presence of PRRS was confirmed by serological detection of antibodies against PRRS virus (PRRSV) and detection of PRRS viral RNA by RT-PCR. Pigs were further divided into three groups of 30: piglets just before weaning (weaners), fatteners and finishers. Blood samples for determining selected blood parameters were collected from the vena cava cranialis. Significantly (P stress might be increased in PRRSV naturally infected pigs, especially in weaners.

  18. Changes in the cellular proteins of pulmonary alveolar macrophage infected with porcine reproductive and respiratory syndrome virus by proteomics analysis.

    Science.gov (United States)

    Zhang, Haiming; Guo, Xin; Ge, Xinna; Chen, Yanhong; Sun, Qixin; Yang, Hanchun

    2009-06-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is known to infect and replicate preferentially in pulmonary alveolar macrophages (PAMs). We applied proteomic approaches to investigate the change in cellular proteins of PAMs infected with PRRSV in vitro. A total of 23 cellular proteins with significant alteration in different courses postinfection were identified. These proteins could be classified into the functions associated with morphogenesis, protein synthesis, metabolism, and stress response and ubiquitin-proteasome pathway. Of the altered proteins, two proteins, heat shock 27 kDa protein (HSP27) and superoxide dismutase 2 (SOD2), involved in stress response or ubiquitin-proteasome pathway were observed to be up-regulated. Our study is the first attempt to analyze the cellular protein profile of PRRSV-infected PAMs by proteomics and provide valuable information for better understanding the function alterations of PAMs induced by PRRSV infection.

  19. Role of transcription regulatory sequence in regulation of gene expression and replication of porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Wang, Chengbao; Meng, Han; Gao, Yujin; Gao, Hui; Guo, Kangkang; Almazan, Fernando; Sola, Isabel; Enjuanes, Luis; Zhang, Yanming; Abrahamyan, Levon

    2017-08-10

    In order to gain insight into the role of the transcription regulatory sequences (TRSs) in the regulation of gene expression and replication of porcine reproductive and respiratory syndrome virus (PRRSV), the enhanced green fluorescent protein (EGFP) gene, under the control of the different structural gene TRSs, was inserted between the N gene and 3'-UTR of the PRRSV genome and EGFP expression was analyzed for each TRS. TRSs of all the studied structural genes of PRRSV positively modulated EGFP expression at different levels. Among the TRSs analyzed, those of GP2, GP5, M, and N genes highly enhanced EGFP expression without altering replication of PRRSV. These data indicated that structural gene TRSs could be an extremely useful tool for foreign gene expression using PRRSV as a vector.

  20. Spatiotemporal analysis of the Porcine Reproductive and Respiratory Syndrome (PRRS) epidemic in Denmark using laboratory submission data

    DEFF Research Database (Denmark)

    Lopes Antunes, Ana Carolina; Toft, Nils; Hisham Beshara Halasa, Tariq

    Porcine reproductive and respiratory syndrome (PRRS) virus infects domestic swine populations causing production losses in many European countries. The virus has two different strains designated as European (EU) and American (US) strain. It has been assumed that 30% of Danish swine herds are sero...... spatiotemporal analysis of serological tests. Records of PRRS serology submissions made from January 2007 to December 2010 stored in the DIANOVA Information Management System at the National Veterinary Institute (DTU Vet) were used in the analysis. Each submission consists in individual blood samples collected....... The herd numbers registered in the laboratory submissions were merged with the Danish Herd Identification System database, in order to obtain the geo-coordinates of the herds. Spatiotemporal analysis was performed, in order to characterize PRRS-EU and PRRSUS distributions for both control levels...

  1. Attempted experimental reproduction of porcine periweaning-failure-to-thrive syndrome using tissue homogenates.

    Science.gov (United States)

    Huang, Yanyun; Harding, John C S

    2014-01-01

    Porcine periweaning failure-to-thrive syndrome (PFTS) is characterized by anorexia and progressive debilitation of newly weaned pigs, of which some also demonstrate repetitive oral behaviour. Although no relevant porcine pathogens have been shown to be causally associated, inoculation of susceptible pigs using tissue homogenates is needed to rule out infectious etiologies. Eight snatched-farrowed porcine-colostrum-deprived (SF-pCD) pigs were inoculated with tissue homogenates made from PFTS-affected pigs orally, or combined orally, intraperitoneally (i.p.) and intramuscularly (i.m.) at day (D) 14 of age (INOC). On D21, i.p. and i.m. inoculation were repeated. Four sham-inoculated pigs served as control (CTRL). Three INOC pigs developed mixed bacterial septicemia between the first and second inoculation. All other pigs survived until termination on D49. Average daily gain (ADG) and the frequencies of diarrhea did not differ between INOC and CTRL pigs D14 and D29. Additionally, the progressive debilitation characteristic of PFTS was not observed in any pig, and repetitive oral behaviour was observed in both groups. In conclusion, PFTS was not experimentally reproduced by the current experimental approach providing evidence that PFTS may not have an infectious etiology.

  2. Attempted experimental reproduction of porcine periweaning-failure-to-thrive syndrome using tissue homogenates.

    Directory of Open Access Journals (Sweden)

    Yanyun Huang

    Full Text Available Porcine periweaning failure-to-thrive syndrome (PFTS is characterized by anorexia and progressive debilitation of newly weaned pigs, of which some also demonstrate repetitive oral behaviour. Although no relevant porcine pathogens have been shown to be causally associated, inoculation of susceptible pigs using tissue homogenates is needed to rule out infectious etiologies. Eight snatched-farrowed porcine-colostrum-deprived (SF-pCD pigs were inoculated with tissue homogenates made from PFTS-affected pigs orally, or combined orally, intraperitoneally (i.p. and intramuscularly (i.m. at day (D 14 of age (INOC. On D21, i.p. and i.m. inoculation were repeated. Four sham-inoculated pigs served as control (CTRL. Three INOC pigs developed mixed bacterial septicemia between the first and second inoculation. All other pigs survived until termination on D49. Average daily gain (ADG and the frequencies of diarrhea did not differ between INOC and CTRL pigs D14 and D29. Additionally, the progressive debilitation characteristic of PFTS was not observed in any pig, and repetitive oral behaviour was observed in both groups. In conclusion, PFTS was not experimentally reproduced by the current experimental approach providing evidence that PFTS may not have an infectious etiology.

  3. Porcine focal symmetrical poliomyelomalacia: experimental reproduction with oral doses of encapsulated sodium selenite.

    Science.gov (United States)

    Wilson, T M; Hammerstedt, R H; Palmer, I S; deLahunta, A

    1988-01-01

    Sodium selenite (encapsulated as doses of 1.4 mg, 2.6 mg and 4.2 mg per kilogram of body weight) was given orally on a daily basis to male weaner pigs, and features of these animals were compared to a control group. Porcine focal symmetrical poliomyelomalacia was produced in all experimental groups between 3 and 20 days after initiation of the treatment. Analysis of blood and several tissues revealed an elevated selenium content for all pigs. Histological lesions in the brain and the cervical lumbar/sacral spinal cord enlargements included endothelial proliferation, neuronal degeneration, microcavitation and glial cell reaction.

  4. Porcine focal symmetrical poliomyelomalacia: experimental reproduction with oral doses of encapsulated sodium selenite.

    OpenAIRE

    Wilson, T.M.; Hammerstedt, R H; Palmer, I S; deLahunta, A

    1988-01-01

    Sodium selenite (encapsulated as doses of 1.4 mg, 2.6 mg and 4.2 mg per kilogram of body weight) was given orally on a daily basis to male weaner pigs, and features of these animals were compared to a control group. Porcine focal symmetrical poliomyelomalacia was produced in all experimental groups between 3 and 20 days after initiation of the treatment. Analysis of blood and several tissues revealed an elevated selenium content for all pigs. Histological lesions in the brain and the cervical...

  5. Evaluation of contact exposure as a method for acclimatizing growing pigs to porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Vashisht, Kapil; Erlandson, Keith R; Firkins, Lawrence D; Zuckermann, Federico A; Goldberg, Tony L

    2008-05-15

    To determine whether 6.5-week-old gilts that have not previously been exposed to porcine reproductive and respiratory syndrome (PRRS) virus can be acclimatized to an endemic strain of the virus by commingling with age-matched gilts inoculated with the endemic PRRS virus strain and whether 10.5-week-old gilts can be acclimatized by commingling with age-matched inoculated or contact-exposed animals. Randomized controlled longitudinal study. 80 gilts seronegative for PRRS on a farm in the Midwestern United States with a history of PRRS. 20 gilts were inoculated with the endemic PRRS virus strain at 6.5 weeks of age (group 1) and were commingled with 20 gilts that were not inoculated (group 2). Four weeks later, the remaining 40 gilts (group 3) were commingled with gilts in groups 1 and 2. Presence of viral RNA in the tonsils, seroconversion rate, serum neutralizing antibody titers, interferon-gamma-mediated cellular immunity, and reproductive outcomes were analyzed. Acclimatization of PRRS virus-naïve pigs was achieved by means of contact exposure at both 6.5 and 10.5 weeks of age. No differences were observed among the 3 groups with respect to development of anti-PRRS virus-specific immune responses or reproductive outcomes. Results suggested that contact exposure of 6.5- to 10.5-week-old pigs that had not previously been exposed to PRRS virus to pigs inoculated with endemic PRRS virus may be an efficient acclimatization strategy for controlling outbreaks on commercial farms on which PRRS is endemic.

  6. Identification of a Divergent Lineage Porcine Pestivirus in Nursing Piglets with Congenital Tremors and Reproduction of Disease following Experimental Inoculation.

    Directory of Open Access Journals (Sweden)

    Bailey L Arruda

    , tracheobronchial lymph node (37 out of 41, and whole blood (19 out of 20. Although the first description of congenital tremors was in 1922, this is the first reported reproduction of congenital tremors following experimental inoculation with a divergent lineage porcine pestivirus. Studies investigating disease mechanism, epidemiology, and diagnostic assay development are needed to better understand the pathophysiology of congenital tremors due to this pestivirus.

  7. Study of the virulence and cross-neutralization capability of recent porcine parvovirus field isolates and vaccine viruses in experimentally infected pregnant gilts.

    Science.gov (United States)

    Zeeuw, E J L; Leinecker, N; Herwig, V; Selbitz, H-J; Truyen, U

    2007-02-01

    The pathogenicity of two recent German field isolates of Porcine parvovirus (PPV-27a and PPV-143a) and two vaccine viruses [PPV-NADL-2 and PPV-IDT (MSV)], which are used for the production of inactivated vaccines, was investigated by inoculation of pregnant sows at day 40 of gestation. Post-infection sera of these sows as well as antisera prepared in rabbits by immunization with the four above-mentioned PPV isolates and with the virulent strain PPV-Challenge (Engl.) were tested for their homologous and heterologous neutralization activities. All antisera had high neutralization activity against the vaccine viruses, the PPV-Challenge (Engl.) virus and PPV-143a, but much lower activity against PPV-27a. These results suggest that PPV-27a represents a new antigenic variant or type of PPV and vaccines based on the established vaccine viruses may not be fully protective against this field isolate. PPV-27a has been characterized based on the amino acid sequences of the capsid protein as a member of a new and distinct PPV cluster (Zimmermann et al., 2006). Interestingly, the homologous neutralizing antibody titres of the sera of all three pigs and both rabbits inoculated or immunized with PPV-27a were 100- to 1000-fold lower than the heterologous titres against any of the other viruses. The low homologous neutralizing antibody titres suggest a possible, yet undefined, immune escape mechanism of this PPV isolate.

  8. Changes in leukocyte subsets of pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome

    OpenAIRE

    Ladinig, Andrea; Gerner, Wilhelm; Saalmüller, Armin; Lunney, Joan K; Ashley, Carolyn; Harding, John CS

    2014-01-01

    International audience; In spite of more than two decades of extensive research, the understanding of porcine reproductive and respiratory syndrome virus (PRRSv) immunity is still incomplete. A PRRSv infection of the late term pregnant female can result in abortions, early farrowings, fetal death, and the birth of weak, congenitally infected piglets. The objectives of the present study were to investigate changes in peripheral blood mononuclear cell populations in third trimester pregnant fem...

  9. An innovative approach to induce cross-protective immunity against porcine reproductive and respiratory syndrome virus in the lungs of pigs through adjuvanted nanotechnology-based vaccination

    OpenAIRE

    Binjawadagi B; Dwivedi V; Manickam C; Ouyang K; Torrelles JB; Renukaradhya GJ

    2014-01-01

    Basavaraj Binjawadagi,1,2 Varun Dwivedi,1 Cordelia Manickam,1,2 Kang Ouyang,1 Jordi B Torrelles,3 Gourapura J Renukaradhya1,21Food Animal Health Research Program, Ohio Agricultural Research and Development Center, 2Department of Veterinary Preventive Medicine, The Ohio State University, Wooster, OH, USA; 3Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH, USAAbstract: Porcine reproductive and respiratory syndrome (PRRS) is an economically devastating resp...

  10. Further evaluation of alternative air-filtration systems for reducing the transmission of Porcine reproductive and respiratory syndrome virus by aerosol

    OpenAIRE

    Dee, Scott A.; Deen, John; Cano, Jean Paul; Batista, Laura; Pijoan, Carlos

    2006-01-01

    The purpose of this study was to compare 4 methods for the reduction of aerosol transmission of Porcine reproductive and respiratory syndrome virus (PRRSV): high-efficiency particulate air (HEPA) filtration, 2×-low-cost filtration, bag filtration, and use of a filter tested against particles derived from dioctylphthalate (DOP). The HEPA-filtration system used a prefilter screen, a bag filter (Eurovent [EU] 8 rating), and a HEPA filter (EU13 rating). The low-cost-filtration system contained mo...

  11. Porcine Reproductive and Respiratory Syndrome Virus Infection Induces Stress Granule Formation Depending on Protein Kinase R-like Endoplasmic Reticulum Kinase (PERK) in MARC-145 Cells

    OpenAIRE

    Zhou, Yanrong; Fang, Liurong; Wang, Dang; Cai, Kaimei; Chen, Huanchun; Xiao, Shaobo

    2017-01-01

    Stress granules (SGs) are sites of mRNA storage that are formed in response to various conditions of stress, including viral infections. Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus that has been devastating the swine industry worldwide since the late 1980s. In this study, we found that infection of PRRSV strain WUH3 (genotype 2 PRRSV) induced stable formation of robust SGs in MARC-145 cells, as demonstrated by the recruitment of marker proteins of SGs, includ...

  12. Heterologous challenge with porcine reproductive and respiratory syndrome (PRRS) vaccine virus: no evidence of reactivation of previous European-type PRRS virus infection

    DEFF Research Database (Denmark)

    Bøtner, Anette; Nielsen, Jens; Oleksiewicz, M.B.

    1999-01-01

    In Denmark, a porcine reproductive and respiratory syndrome virus (PRRSV) control programme, comprising vaccination of seropositive herds with a live American type PRRSV vaccine, was started in 1996. In several of these herds, spread of vaccine virus from vaccinated 3-18 week old pigs to non...... in previously European PRRSV infected pigs after challenge with the vaccine strain seems to be the result of a boosting effect on the immune system, induced by the heterologous vaccine PRRSV strain....

  13. A Dimerization-Dependent Mechanism Drives the Endoribonuclease Function of Porcine Reproductive and Respiratory Syndrome Virus nsp11.

    Science.gov (United States)

    Shi, Yuejun; Li, Youwen; Lei, Yingying; Ye, Gang; Shen, Zhou; Sun, Limeng; Luo, Rui; Wang, Dang; Fu, Zhen F; Xiao, Shaobo; Peng, Guiqing

    2016-05-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) RNA endoribonuclease nsp11 belongs to the XendoU superfamily and plays a crucial role in arterivirus replication. Here, we report the first crystal structure of the arterivirus nsp11 protein from PRRSV, which exhibits a unique structure and assembles into an asymmetric dimer whose structure is completely different from the hexameric structure of coronavirus nsp15. However, the structures of the PRRSV nsp11 and coronavirus nsp15 catalytic domains were perfectly superimposed, especially in the "active site loop" (His129 to His144) and "supporting loop" (Val162 to Thr179) regions. Importantly, our biochemical data demonstrated that PRRSV nsp11 exists mainly as a dimer in solution. Mutations of the major dimerization site determinants (Ser74 and Phe76) in the dimerization interface destabilized the dimer in solution and severely diminished endoribonuclease activity, indicating that the dimer is the biologically functional unit. In the dimeric structure, the active site loop and supporting loop are packed against one another and stabilized by monomer-monomer interactions. These findings may help elucidate the mechanism underlying arterivirus replication and may represent great potential for the development of antiviral drugs. Porcine reproductive and respiratory syndrome virus (PRRSV) is a member of the family Arteriviridae, order Nidovirales PRRSV is a major agent of respiratory diseases in pigs, causing tremendous economic losses to the swine industry worldwide. The PRRSV nsp11 endoribonuclease plays a vital role in arterivirus replication, but its precise roles and mechanisms of action are poorly understood. Here, we report the first dimeric structure of the arterivirus nsp11 from PRRSV at 2.75-Å resolution. Structural and biochemical experiments demonstrated that nsp11 exists mainly as a dimer in solution and that nsp11 may be fully active as a dimer. Mutagenesis and structural analysis revealed Nendo

  14. Usefulness of bovine and porcine IVM/IVF models for reproductive toxicology

    NARCIS (Netherlands)

    Santos, Regiane R; Schoevers, Eric J; Roelen, Bernard Aj

    2014-01-01

    Women presenting fertility problems are often helped by Assisted Reproductive Techniques (ART), such as in vitro fertilization (IVF) programs. However, in many cases the etiology of the in/subfertility remains unknown even after treatment. Although several aspects should be considered when assisting

  15. Molecular evolution of type 2 porcine reproductive and respiratory syndrome viruses circulating in Vietnam from 2007 to 2015.

    Science.gov (United States)

    Do, Hai Quynh; Trinh, Dinh Thau; Nguyen, Thi Lan; Vu, Thi Thu Hang; Than, Duc Duong; Van Lo, Thi; Yeom, Minjoo; Song, Daesub; Choe, SeEun; An, Dong-Jun; Le, Van Phan

    2016-11-17

    Porcine respiratory and reproductive syndrome (PRRS) virus is one of the most economically significant pathogens in the Vietnamese swine industry. ORF5, which participates in many functional processes, including virion assembly, entry of the virus into the host cell, and viral adaptation to the host immune response, has been widely used in molecular evolution and phylogeny studies. Knowing of molecular evolution of PRRSV fields strains might contribute to PRRS control in Vietnam. The results showed that phylogenetic analysis indicated that all strains belonged to sub-lineages 8.7 and 5.1. The nucleotide and amino acid identities between strains were 84.5-100% and 82-100%, respectively. Furthermore, the results revealed differences in nucleotide and amino acid identities between the 2 sub-lineage groups. N-glycosylation prediction identified 7 potential N-glycosylation sites and 11 glycotypes. Analyses of the GP5 sequences, revealed 7 sites under positive selective pressure and 25 under negative selective pressure. Phylogenetic analysis based on ORF5 sequence indicated the diversity of PRRSV in Vietnam. Furthermore, the variance of N-glycosylation sites and position under selective pressure were demonstrated. This study expands existing knowledge on the genetic diversity and evolution of PRRSV in Vietnam and assists the effective strategies for PRRS vaccine development in Vietnam.

  16. The potential adjuvanticity of quaternized chitosan hydrogel based microparticles for porcine reproductive and respiratory syndrome virus inactivated vaccine.

    Science.gov (United States)

    Wang, Yue-Qi; Liu, Yan; Wang, Yu-Xia; Wu, Ya-Jun; Jia, Pei-Yuan; Shan, Jun-Jie; Wu, Jie; Ma, Guang-Hui; Su, Zhi-Guo

    2016-10-01

    Infectious diseases possess a big threat to the livestock industry worldwide. Currently, inactivated veterinary vaccines have attracted much attention to prevent infection due to their safer profile compared to live attenuated vaccine. However, its intrinsic poor immunogenicity demands the incorporation of an adjuvant. Mineral oil based adjuvant (Montanide™ ISA206) was usually used to potentiate the efficacy of veterinary vaccines. However, ISA206 could not induce robust cellular immune responses, which was very important in controlling virus replication and clearing the infected cells. Moreover, mineral oil would result in severe side effects. To improve both the humoral and cellular immune responses of porcine reproductive and respiratory syndrome virus (PRRSV) inactivated vaccine, we developed pH-sensitive and size-controllable quaternized chitosan hydrogel microparticles (Gel MPs) without using chemical cross linking agent. Gel MPs, ionic cross-linked with glycerophosphate (GP), were biocompatible and could efficiently adsorb the inactivated PRRSV vaccine with a loading capacity of 579.05μg/mg. After intramuscular immunization in mice, results suggested that Gel MPs elicited significantly higher cell-mediated immune responses and comparable humoral immune responses compared to ISA 206. Regarding the biocompatibility, safety and effectiveness, Gel MPs would be a promising candidate to enhance the efficacy of veterinary vaccine. Copyright © 2016. Published by Elsevier B.V.

  17. Chimeric porcine reproductive and respiratory syndrome virus containing shuffled multiple envelope genes confers cross-protection in pigs.

    Science.gov (United States)

    Tian, Debin; Ni, Yan-Yan; Zhou, Lei; Opriessnig, Tanja; Cao, Dianjun; Piñeyro, Pablo; Yugo, Danielle M; Overend, Christopher; Cao, Qian; Lynn Heffron, C; Halbur, Patrick G; Pearce, Douglas S; Calvert, Jay G; Meng, Xiang-Jin

    2015-11-01

    The extensive genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) strains is a major obstacle for vaccine development. We previously demonstrated that chimeric PRRSVs in which a single envelope gene (ORF3, ORF4, ORF5 or ORF6) was shuffled via DNA shuffling had an improved heterologous cross-neutralizing ability. In this study, we incorporate all of the individually-shuffled envelope genes together in different combinations into an infectious clone backbone of PRRSV MLV Fostera(®) PRRS. Five viable progeny chimeric viruses were rescued, and their growth characteristics were characterized in vitro. In a pilot pig study, two chimeric viruses (FV-SPDS-VR2,FV-SPDS-VR5) were found to induce cross-neutralizing antibodies against heterologous strains. A subsequent vaccination/challenge study in 72 pigs revealed that chimeric virus FV-SPDS-VR2 and parental virus conferred partial cross-protection when challenged with heterologous strains NADC20 or MN184B. The results have important implications for future development of an effective PRRSV vaccine that confers heterologous protection. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Chemical constituents from Chirita longgangensis var. hongyao with inhibitory activity against porcine respiratory and reproductive syndrome virus

    Energy Technology Data Exchange (ETDEWEB)

    Su, Yao; Wang, Yue-Hu; Tan, Ying; Yang, Jun; Liu, Hong-Xin; Gu, Wei; Long, Chun-Lin, E-mail: long@mail.kib.ac.cn [Key Laboratory of Economic Plants and Biotechnology, Kunming Institute of Botany, Chinese Academy of Sciences (China); Bi, Jun-Long; Yin, Ge-Fen, E-mail: yingefen383@sohu.com [College of Animal Science and Technology, Yunnan Agricultural University (China)

    2012-10-15

    Two new quinonoids chiritalone A and B, and a new neolignan 7'E-4,9-dihydroxy- 3,3',5'-trimethoxy-8,4'-oxyneolign-7'-en-9'-al, along with known (-)-8-hydroxy-{alpha}-dunnione, digiferruginol, 2,5-dimethoxy-1,4-benzoquinone and hederagenin, were isolated from the stems of Chirita longgangensis var. hongyao. The structures of the new compounds were elucidated by detailed analysis from NMR (nuclear magnetic resonance) and MS (mass spectrometry) data, and the absolute configuration of chiritalone A was determined by single crystal X-ray diffraction analysis using the Flack parameter. The inhibitory activity of compounds against porcine respiratory and reproductive syndrome virus (PRRSV) was measured by the cytopathic effect (CPE) method. Digiferruginol and hederagenin showed weak effect on PRRSV with an IC{sub 50} value of 80.5 {+-} 16.9 {mu}mol L{sup -1} (SI = 19.9) and 43.2 {+-} 7.4 {mu}mol L{sup -1} (SI = 13.1), respectively. (author)

  19. Interactome profile of the host cellular proteins and the nonstructural protein 2 of porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Wang, Li; Zhou, Lei; Zhang, Han; Li, Yan; Ge, Xinna; Guo, Xin; Yu, Kangzhen; Yang, Hanchun

    2014-01-01

    The nonstructural protein 2 (NSP2) is considered to be one of crucial viral proteins in the replication and pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV). In the present study, the host cellular proteins that interact with the NSP2 of PRRSV were immunoprecipitated with anti-Myc antibody from the MARC-145 cells infected by a recombinant PRRSV with 3xMyc tag insertion in its NSP2-coding region, and then 285 cellular proteins interacting with NSP2 were identified by LC-MS/MS. The Gene Ontology and enriched KEGG Pathway bioinformatics analyses indicated that the identified proteins could be assigned to different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with infectious disease, translation, immune system, nervous system and signal transduction. Two interested cellular proteins-BCL2-associated athanogene 6 (BAG6) and apoptosis-inducing factor 1 (AIF1) which may involve in transporting of NSP2 to Endoplasmic reticulum (ER) or PRRSV-driven apoptosis were validated by Western blot. The interactome data between PRRSV NSP2 and cellular proteins contribute to the understanding of the roles of NSP2 in the replication and pathogenesis of PRRSV, and also provide novel cellular target proteins for elucidating the associated molecular mechanisms of the interaction of host cellular proteins with viral proteins in regulating the viral replication.

  20. Interactome profile of the host cellular proteins and the nonstructural protein 2 of porcine reproductive and respiratory syndrome virus.

    Directory of Open Access Journals (Sweden)

    Li Wang

    Full Text Available The nonstructural protein 2 (NSP2 is considered to be one of crucial viral proteins in the replication and pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV. In the present study, the host cellular proteins that interact with the NSP2 of PRRSV were immunoprecipitated with anti-Myc antibody from the MARC-145 cells infected by a recombinant PRRSV with 3xMyc tag insertion in its NSP2-coding region, and then 285 cellular proteins interacting with NSP2 were identified by LC-MS/MS. The Gene Ontology and enriched KEGG Pathway bioinformatics analyses indicated that the identified proteins could be assigned to different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with infectious disease, translation, immune system, nervous system and signal transduction. Two interested cellular proteins-BCL2-associated athanogene 6 (BAG6 and apoptosis-inducing factor 1 (AIF1 which may involve in transporting of NSP2 to Endoplasmic reticulum (ER or PRRSV-driven apoptosis were validated by Western blot. The interactome data between PRRSV NSP2 and cellular proteins contribute to the understanding of the roles of NSP2 in the replication and pathogenesis of PRRSV, and also provide novel cellular target proteins for elucidating the associated molecular mechanisms of the interaction of host cellular proteins with viral proteins in regulating the viral replication.

  1. Reproduction in porcine circovirus type 2 (PCV2) seropositive gilts inseminated with PCV2b spiked semen.

    Science.gov (United States)

    Sarli, Giuseppe; Morandi, Federico; Panarese, Serena; Bacci, Barbara; Ferrara, Domenico; Bianco, Carlo; Fusaro, Laura; Bacci, Maria Laura; Galeati, Giovanna; Dottori, Michele; Bonilauri, Paolo; Lelli, Davide; Leotti, Giorgio; Vila, Thais; Joisel, Francois; Allan, Gordon; Benazzi, Cinzia; Ostanello, Fabio

    2012-08-31

    Since 1999, field evidence of transplacental infection by porcine circovirus type 2 (PCV2) and reproductive failure has been reported in pigs. The objective of this study was to evaluate the clinical and pathological consequences of PCV2 infection in conventional PCV2-seropositive gilts by insemination with PCV2b-spiked semen. Six PCV2 seropositive gilts were inseminated with PCV2b-supplemented semen (infected) and three animals with semen and cell culture medium (controls). Only three out of the six infected animals were pregnant by ultrasonography on day 29 after insemination, while two out of the three controls were pregnant. One control gilt aborted on day 23 after insemination but not due to PVC2. Viraemia was demonstrated in four out of six infected and in one control gilt that became infected with PCV2a. Anti-PCV2 antibody titres showed dynamic variations in the infected group throughout the study. Among infected gilts, the animal with the lowest anti-PCV2 titre (1/100) at the beginning of the experiment and another that reached a similar low value during the experiment showed evident seroconversion over time and had also PCV2 positive foetuses. One placenta displayed mild focal necrosis of the chorionic epithelium positively stained by immunohistochemistry for PCV2 antigen. PCV2-seropositive gilts can be infected with PCV2 after intrauterine exposure and low maternal antibody titre may increase the probability of a foetal infection.

  2. The assessment of efficacy of porcine reproductive respiratory syndrome virus inactivated vaccine based on the viral quantity and inactivation methods

    Directory of Open Access Journals (Sweden)

    Lee Byeongchun

    2011-06-01

    Full Text Available Abstract Background There have been many efforts to develop efficient vaccines for the control of porcine reproductive and respiratory syndrome virus (PRRSV. Although inactivated PRRSV vaccines are preferred for their safety, they are weak at inducing humoral immune responses and controlling field PRRSV infection, especially when heterologous viruses are involved. Results In all groups, the sample to positive (S/P ratio of IDEXX ELISA and the virus neutralization (VN titer remained negative until challenge. While viremia did not reduce in the vaccinated groups, the IDEXX-ELISA-specific immunoglobulin G increased more rapidly and to significantly greater levels 7 days after the challenge in all the vaccinated groups compared to the non-vaccinated groups (p 6 PFU/mL PRRSV vaccine-inoculated and binary ethylenimine (BEI-inactivated groups 22 days after challenge (p Conclusions The inactivated vaccine failed to show the humoral immunity, but it showed different immune response after the challenge compared to mock group. Although the 106 PFU/mL-vaccinated and BEI-inactivated groups showed significantly greater VN titers 22 days after challenge, all the groups were already negative for viremia.

  3. Porcine reproductive and respiratory syndrome virus (PRRSV in GB pig herds: farm characteristics associated with heterogeneity in seroprevalence

    Directory of Open Access Journals (Sweden)

    Medley Graham F

    2008-11-01

    Full Text Available Abstract Background The between- and within-herd variability of porcine reproductive and respiratory syndrome virus (PRRSV antibodies were investigated in a cross-sectional study of 103 British pig herds conducted 2003–2004. Fifty pigs from each farm were tested for anti-PRRSV antibodies using ELISA. A binomial logistic model was used to investigate management risks for farms with and without pigs with PRRSV antibodies and multilevel statistical models were used to investigate variability in pigs' log ELISA IRPC (relative index × 100 in positive herds. Results Thirty-five herds (34.0% were seronegative, 41 (39.8% were seropositive and 27 (26.2% were vaccinated. Herds were more likely to be seronegative if they had Conclusion These patterns are consistent with PRRSV failing to persist indefinitely on some infected farms, with fadeout more likely in smaller herds with little/no reintroduction of infectious stock. Persistence of infection may be associated with large herds in pig-dense regions with repeated reintroduction.

  4. Strain predominance following exposure of vaccinated and naive pregnant gilts to multiple strains of porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Lager, Kelly M; Mengeling, William L; Wesley, Ronald D

    2003-05-01

    Two studies were performed in order to test the relative ability of different strains of porcine reproductive and respiratory syndrome virus (PRRSV) to replicate and cross the placental barrier in pregnant gilts. Study 1 comprised 6 nonvaccinated gilts. Study 2 comprised 8 nonvaccinated gilts and 12 gilts that were vaccinated twice before conception. On, or about, gestation day 90 all gilts were simultaneously exposed to 20 field strains of PRRSV (all strains were distinguishable by restriction fragment length polymorphism (RFLP) patterns). Gilts of study 1 were euthanized on day 7 postpartum. Gilts of study 2 were euthanized on, or about, gestation day 111. All gilts, pigs, and fetuses were tested for the presence and type of strain of PRRSV. Of 128 samples shown to contain PRRSV, 118 contained a single strain, 4 contained 2 strains, and 2 contained a strain or strains for which the RFLP pattern was undecipherable. Only 8 of the 20 strains were isolated from nonvaccinated gilts and their litters. And only 2 of the 20 strains (notably 2 of the same strains isolated from nonvaccinated gilts and their litters), were isolated from vaccinated gilts and their litters. Moreover, 1 of the 2 strains accounted for most (31 of 37; 84%) of the isolates from the vaccinated group. Collectively these results indicate that strains differ in their ability to replicate in pregnant gilts and cross the placental barrier. And they suggest that maternal immunity, although sometimes insufficient to prevent transplacental infection, can exert additional selective pressure.

  5. Detection and typing of highly pathogenic porcine reproductive and respiratory syndrome virus by multiplex real-time rt-PCR.

    Directory of Open Access Journals (Sweden)

    Kerstin Wernike

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS causes economic losses in the pig industry worldwide, and PRRS viruses (PRRSV are classified into the two distinct genotypes "North American (NA, type 2" and "European (EU, type 1". In 2006, a highly pathogenic NA strain of PRRSV (HP-PRRSV, characterized by high fever as well as high morbidity and mortality, emerged in swine farms in China. Therefore, a real-time reverse transcription polymerase chain reaction (RT-qPCR assay specific for HP-PRRSV was developed and combined with type 1- and type 2-specific RT-qPCR systems. Furthermore, an internal control, based on a heterologous RNA, was successfully introduced. This final multiplex PRRSV RT-qPCR, detecting and typing PRRSV, had an analytical sensitivity of less than 200 copies per µl for the type 1-assay and 20 copies per µl for the type 2- and HP assays and a high diagnostic sensitivity. A panel of reference strains and field isolates was reliably detected and samples from an animal trial with a Chinese HP-PRRS strain were used for test validation. The new multiplex PRRSV RT-qPCR system allows for the first time the highly sensitive detection and rapid differentiation of PRRSV of both genotypes as well as the direct detection of HP-PRRSV.

  6. ICAM-1-dependent and ICAM-1-independent neutrophil lung infiltration by porcine reproductive and respiratory syndrome virus infection.

    Science.gov (United States)

    Liu, Jie; Hou, Make; Yan, Meiping; Lü, Xinhui; Gu, Wei; Zhang, Songlin; Gao, Jianfeng; Liu, Bang; Wu, Xiaoxiong; Liu, Guoquan

    2015-08-01

    Neutrophils are innate immune cells that play a crucial role in the first line of host defense. It is also known that neutrophil lung recruitment and infiltration may cause lung injury. The roles of neutrophils in virus infection-induced lung injury are not clear. We explore the mechanisms of neutrophil lung infiltration and the potential biomarkers for lung injury in a swine model of lung injury caused by natural or experimental porcine reproductive and respiratory syndrome virus (PRRSV) infection. Neutrophil lung infiltration was determined by measurement of myeloperoxidase expression and enzyme activity of lung tissues. Myeloperoxidase expression and enzyme activity were dramatically increased in the naturally and experimentally infected lung tissues. Chemokine analysis by quantitative PCR and ELISA showed that IL-8 expression was increased in both infections, while monocyte chemoattractant protein-1 expression was increased only in experimentally infected lung tissues. Expression of the cell adhesion molecules VCAM-1 and ICAM-1 was measured by quantitative PCR and Western blotting. VCAM-1 expression was increased in experimentally and naturally infected lungs, whereas ICAM-1 expression was increased only in the naturally infected lung samples. Our results suggest that neutrophil lung infiltrations in the infected animals are both ICAM-1- and -independent and that combined expression of VCAM-1 and IL-8 may serve as the biomarker for lung injury induced by virus infection. Copyright © 2015 the American Physiological Society.

  7. A highly pathogenic porcine reproductive and respiratory syndrome virus candidate vaccine based on Japanese encephalitis virus replicon system.

    Science.gov (United States)

    Hu, Pingsheng; Chen, Xiaoming; Huang, Lihong; Liu, Shukai; Zang, Fuyu; Xing, Jinchao; Zhang, Youyue; Liang, Jiaqi; Zhang, Guihong; Liao, Ming; Qi, Wenbao

    2017-01-01

    In the swine industry, porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease which causes heavy economic losses worldwide. Effective prevention and disease control is an important issue. In this study, we described the construction of a Japanese encephalitis virus (JEV) DNA-based replicon with a cytomegalovirus (CMV) promoter based on the genome of Japanese encephalitis live vaccine virus SA14-14-2, which is capable of offering a potentially novel way to develop and produce vaccines against a major pathogen of global health. This JEV DNA-based replicon contains a large deletion in the structural genes (C-prM-E). A PRRSV GP5/M was inserted into the deletion position of JEV DNA-based replicons to develop a chimeric replicon vaccine candidate for PRRSV. The results showed that BALB/c mice models with the replicon vaccines pJEV-REP-G-2A-M-IRES and pJEV-REP-G-2A-M stimulated antibody responses and induced a cellular immune response. Analysis of ELSA data showed that vaccination with the replicon vaccine expressing GP5/M induced a better antibodies response than traditional DNA vaccines. Therefore, the results suggested that this ectopic expression system based on JEV DNA-based replicons may represent a useful molecular platform for various biological applications, and the JEV DNA-based replicons expressing GP5/M can be further developed into a novel, safe vaccine candidate for PRRS.

  8. Reproduction in porcine circovirus type 2 (PCV2 seropositive gilts inseminated with PCV2b spiked semen

    Directory of Open Access Journals (Sweden)

    Sarli Giuseppe

    2012-08-01

    Full Text Available Abstract Background Since 1999, field evidence of transplacental infection by porcine circovirus type 2 (PCV2 and reproductive failure has been reported in pigs. The objective of this study was to evaluate the clinical and pathological consequences of PCV2 infection in conventional PCV2-seropositive gilts by insemination with PCV2b-spiked semen. Results Six PCV2 seropositive gilts were inseminated with PCV2b-supplemented semen (infected and three animals with semen and cell culture medium (controls. Only three out of the six infected animals were pregnant by ultrasonography on day 29 after insemination, while two out of the three controls were pregnant. One control gilt aborted on day 23 after insemination but not due to PVC2. Viraemia was demonstrated in four out of six infected and in one control gilt that became infected with PCV2a. Anti-PCV2 antibody titres showed dynamic variations in the infected group throughout the study. Among infected gilts, the animal with the lowest anti-PCV2 titre (1/100 at the beginning of the experiment and another that reached a similar low value during the experiment showed evident seroconversion over time and had also PCV2 positive foetuses. One placenta displayed mild focal necrosis of the chorionic epithelium positively stained by immunohistochemistry for PCV2 antigen. Conclusions PCV2-seropositive gilts can be infected with PCV2 after intrauterine exposure and low maternal antibody titre may increase the probability of a foetal infection.

  9. Epidemiological investigations of the introduction of porcine reproductive and respiratory syndrome virus in Chile, 2013-2015.

    Directory of Open Access Journals (Sweden)

    Víctor Neira

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS is endemic in most pork producing countries. In Chile, eradication of PRRS virus (PRRSV was successfully achieved in 2009 as a result of the combined efforts of producers and the animal health authorities. In October 2013, after several years without detecting PRRSV under surveillance activities, suspected cases were confirmed on a commercial swine farm. Here, we describe the PRRS epidemic in Chile between October 2013 and April 2015, and we studied the origins and spread of PRRSV throughout the country using official surveillance data and Bayesian phylogenetic analysis. Our results indicate that the outbreaks were caused by a PRRSV closely related to viruses present in swine farms in North America, and different from the strain that circulated in the country before 2009. Using divergence time estimation analysis, we found that the 2013-2015 PRRSV may have been circulating in Chile for at least one month before the first detection. A single strain of PRRSV spread into a limited number of commercial and backyard swine farms. New infections in commercial systems have not been reported since October 2014, and eradication is underway by clearing the disease from the few commercial and backyard farms that remain positive. This is one of the few documented experiences of PRRSV introduction into a disease-free country.

  10. The diversity of Porcine Reproductive and Respiratory Syndrome Virus Type 1 and 2 in Denmark

    DEFF Research Database (Denmark)

    Kvisgaard, Lise Kirstine; Hjulsager, Charlotte Kristiane; Kristensen, Charlotte Sonne

    Both Type 1 and Type 2 PRRS viruses are circulating among Danish pigs. The first appearance of Type 1 PRRSV in Denmark was in 1992 whereas the Type 2 PRRSV was introduced in 1996 after the use of a live attenuated vaccine that reverted to virulence. Since then, vaccination to control the disease...... strains were sequenced. Denmark exports more than 50.000 living pigs each month. A portion of these pigs inevitably harbor PRRSV. Thus, the diversity of PRRSV in Denmark is of interest to other countries besides Denmark. The main objective of the present study was to close the gap in knowledge...... of the results showed that the Type 1 strains all belonged to subtype 1. Based on the ORF5 sequences, the Danish Type 1 viruses clustered into two groups. These two groups shared 84 % to 92 % and 94 % to 99% nucleotide identity to the Lelystad virus, respectively. The sequenced Type 2 viruses showed...

  11. Splenic CD163(+) macrophages as targets of porcine reproductive and respiratory virus: Role of Siglecs.

    Science.gov (United States)

    Yuste, María; Fernández-Caballero, Teresa; Prieto, Cinta; Álvarez, Belén; Martínez-Lobo, Javier; Simarro, Isabel; Castro, José María; Alonso, Fernando; Ezquerra, Ángel; Domínguez, Javier; Revilla, Concepción

    2017-01-01

    CD169 and CD163 have been involved in the process of PRRS virus attachment and infection in macrophages, although recent studies have challenged the requirement for CD169. In addition to CD169, macrophages express other siglecs, whose role in PRRS virus infection is so far unknown. Splenic CD163(+) macrophages express Siglec-3 and Siglec-5 but almost undetectable levels of CD169. Hence, we considered this cell population appropriate for analysing the role of these siglecs in the attachment and internalization of PRRS virus into macrophages. PRRS virus replicated efficiently in these macrophages, yielding even higher titres than in alveolar macrophages. Besides, a recombinant protein consisting in the ectodomain of porcine Siglec-3 fused to the Fc fragment of human IgG1 (Siglec3-Fc) was able to bind PRRS virus, while binding to Siglec-5-Fc was inconsistent. Antibodies to CD169 but not to Siglec-3 or Siglec-5 blocked the binding and infection of PRRS virus on alveolar macrophages. Unexpectedly, our antibody to CD169 also blocked the binding of PRRS virus to splenic CD163(+) macrophages, whereas antibodies to Siglec-3 or Siglec-5 had no effect. These results show that very low levels of CD169 expression are enough to support the attachment and internalization of PRRS virus into macrophages, whereas Siglec-3 and Siglec-5 do not seem to contribute to the virus entry in these cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Reproduction of porcine proliferative enteropathy with pure cultures of ileal symbiont intracellularis.

    Science.gov (United States)

    McOrist, S; Jasni, S; Mackie, R A; MacIntyre, N; Neef, N; Lawson, G H

    1993-01-01

    Porcine proliferative enteropathy is consistently associated with the presence of intracellular curved bacteria in epithelial cells in affected portions of intestine. Two strains of these intracellular bacteria were cultured in a cell culture system with rat enterocytes (IEC-18) and passaged several times and used as oral inocula for 14 gnotobiotic and 8 conventional pigs. DNA and immunological studies had identified these bacteria as belonging to a new taxon, Ileal symbiont (IS) intracellularis. Conventional pigs dosed with approximately 3.7 x 10(6) of these organisms passaged six times in cell culture developed severe lesions of proliferative enteropathy in the ileum. Other conventional pigs dosed with a lower titer or with organisms passaged 13 times developed moderate and minor lesions, respectively. All gnotobiotic pigs dosed with organisms failed to develop lesions. Control pigs, eight conventional and two gnotobiotic, dosed with diluent, uninfected cell material or left undosed failed to develop lesions also. Reisolation of IS intracellularis and demonstration of the organism in mucosal and fecal samples only occurred in conventional pigs dosed with organisms. Gnotobiotic pigs lacking a normal intestinal flora have not been shown to be colonized by the organism. Seroconversion to IS intracellularis or mucosal infiltration by inflammatory cells was not observed in experimentally affected pigs, confirming the weak immune response characteristic of the natural disease. These results support the identification of IS intracellularis as an etiological agent of proliferative enteropathy in pigs. Images PMID:8406817

  13. Genotype 2 strains of porcine reproductive and respiratory syndrome virus dysregulate alveolar macrophage cytokine production via the unfolded protein response.

    Science.gov (United States)

    Chen, Wei-Yu; Schniztlein, William M; Calzada-Nova, Gabriela; Zuckermann, Federico A

    2017-10-25

    Porcine reproductive and respiratory syndrome virus (PRRSV) infects alveolar macrophages (AMΦ) causing dysregulated interferon (IFN)-α and tumor necrosis factor (TNF)-α production through a mechanism(s) yet to be resolved. Here, we show that AMΦ infected with PRRSV secreted a reduced quantity of IFN-α following the cell exposure to synthetic dsRNA. This reduction did not correlate with reduced IFNA1 gene transcription. Rather, it coincided with two events that occurred late during infection and were indicative of translational attenuation, specifically, the activation of eukaryotic translation initiation factor 2α (eIF2α), and the appearance of stress granules. Notably, the typical rapid production of TNF-α by AMΦ exposed to lipopolysaccharide (LPS) was suppressed or enhanced by PRRSV depending on when the LPS exposure occurred after virus infection. If exposure was delayed until 6 h post-infection (hpi) so that the development of the cytokine response coincided with the time in which phosphorylation of eIF2α by the stress sensor PERK (protein kinase RNA (PKR)-like ER kinase) occurred, inhibition of TNF-α production was observed. However, if LPS exposure occurred at 2 hpi, prior to a detectable onset of eIF2α phosphorylation, a synergistic response was observed due to the earlier NF-κB activation via the stress sensor IRE1α (inositol-requiring kinase 1α). These results suggest that the asynchronous actions of two branches of the unfolded protein response (UPR), namely IRE1α, and PERK, activated by ER stress resulting from the virus infection, are associated with enhancement or suppression of TNF-α production, respectively.IMPORTANCE The activation of AMΦ is controlled by its microenvironment to deter excessive pro-inflammatory cytokine responses to microbes that could impair lung function. However, viral pneumonias frequently become complicated by secondary bacterial infections triggering severe inflammation, lung dysfunction, and death. Although

  14. Occurrence of swine salmonellosis in postweaning multisystemic wasting syndrome (PMWS) affected pigs concurrently infected with porcine reproduction and respiratory syndrome virus (PRRSV).

    Science.gov (United States)

    Murakami, Satoshi; Ogawa, Akihiro; Kinoshita, Tomohide; Matsumoto, Atsuko; Ito, Noriko; Nakane, Takashi

    2006-04-01

    Fourteen diseased pigs from four farms in which there had been an outbreak of salmonellosis were investigated. Granulomatous inflammation with depletion of lymphocytes was observed in the swollen lymph nodes in these pigs. Antigens to porcine circovirus type 2 (PCV2) were immunolabeled in the lesions along with detection of viral DNA as PCV2 by polymerase chain reaction (PCR). In addition, antigens to porcine reproductive respiratory syndrome virus (PRRSV) were immunodetected in the lungs and Salmonella Choleraesuis was isolated from the affected pigs. The nine salmonellosis affected pigs, five (55.6%) with salmonellosis and PMWS concurrently infected with PRRSV were much higher than those infected with salmonellosis and PMWS (22.2%) or with salmonellosis and PPPRV (22.2%).

  15. Experimental inoculation of late term pregnant sows with a field isolate of porcine reproductive and respiratory syndrome vaccine-derived virus

    DEFF Research Database (Denmark)

    Nielsen, Jens; Bøtner, Anette; Bille-Hansen, Vivi

    2002-01-01

    The use of a live attenuated porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in piglets has been associated with reproductive disorders in non-vaccinated sows. Vaccine-derived virus (VDV) has been isolated from foctuses, stillborn pigs, and dead: piglets, indicating...... that the live vaccine spread from vaccinated piglets to non-vaccinated sows, and that the virus might be implicated in the severe reproductive problems observed. In the present study, one such VDV isolate was used to experimentally infect pregnant sows in the last trimester. The chosen isolate, which had more...... than 99.6% identity to the attenuated vaccine virus, originated from the lungs of a stillborn pig from a swine herd with a sudden high level of stillborn pigs and increased piglet mortality in the nursing period. Intranasal inoculation of sows with the virus isolate resulted in congenital infection...

  16. Genetic and biological characterization of a Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2)causing significant clinical disease in the field

    DEFF Research Database (Denmark)

    Kvisgaard, Lise Kirstine; Larsen, Lars Erik; Hjulsager, Charlotte Kristiane

    2017-01-01

    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....... 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...

  17. Genetic diversity of porcine reproductive and respiratory syndrome virus in Thailand and Southeast Asia from 2008 to 2013.

    Science.gov (United States)

    Jantafong, Tippawan; Sangtong, Pradit; Saenglub, Wimontiane; Mungkundar, Chatthapon; Romlamduan, Narin; Lekchareonsuk, Chalermpol; Lekcharoensuk, Porntippa

    2015-04-17

    Porcine reproductive and respiratory syndrome virus (PRRSV) affects the swine industry worldwide. Annual surveillances taken from 2008 to 2013 revealed a 13.86% prevalence of PRRSVs in swine populations in Thailand. The selected positive samples were genetically characterized based on global systems and phylogenetic trees that were constructed using 967 ORF5 samples from this study, the collective sequences from Thailand and Southeast Asia and reference sequences. The results showed that both types I and II have been circulating in Thai swine and that genotype II was more prevalent than genotype I. Only type II was found in other countries in Southeast Asia. Type I PRRSVs from Thailand are clustered in subtype 1, clades A, D and H. Type II PRRSVs are topologically classified in lineage 1 and sublineages 5.1, 5.2 and 8.7, of which sublineage 8.7 was predominant, especially after 2010. PRRSVs in sublineage 8.7 are divided into two groups: classical NA and HP-PRRSV. An analysis of all HP-PRRSVs in Southeast Asia revealed four separate clades--A (SX2009-like), B (09HEN1-like), JXA1-like and GXFCH08-like--reflecting four different introductions of these viruses into Thailand, Lao PDR, Cambodia and Vietnam. HP-PRRSV first appeared in Thailand and Cambodia in 2008, 2 years before the first epidemic outbreaks. Recently, the genetics of PRRSVs in Southeast Asia have become more diverse. Thus, PRRSV genetics must be continually characterized and phylogenetically analyzed using global systematic classifications to provide annual genetic information for PRRS control and vaccine selection. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Applications of Bayesian Phylodynamic Methods in a Recent U.S. Porcine Reproductive and Respiratory Syndrome Virus Outbreak

    Directory of Open Access Journals (Sweden)

    Mohammad A. Alkhamis

    2016-02-01

    Full Text Available Classical phylogenetic methods such as neighbor-joining or maximum likelihood trees, provide limited inferences about the evolution of important pathogens and ignore important evolutionary parameters and uncertainties, which in turn limits decision making related to surveillance, control and prevention resources. Bayesian phylodynamic models have recently been used to test research hypothesis related to evolution of infectious agents. However, few studies have attempted to model the evolutionary dynamics of porcine reproductive and respiratory syndrome virus (PRRSV and, to the authors’ knowledge, no attempt has been made to use large volumes of routinely collected data, sometimes referred to as big data, in the context of animal disease surveillance. The objective of this study was to explore and discuss the applications of Bayesian phylodynamic methods for modeling the evolution and spread of a notable 1-7-4 RFLP-type PRRSV between 2014 and 2015. A convenience sample of 288 ORF5 sequences was collected from 5 swine production systems in the United States between September 2003 and March 2015. Using coalescence and discrete trait phylodynamic models, we were able to infer population growth and demographic history of the virus, identified the most likely ancestral system (root state posterior probability = 0.95 and revealed significant dispersal routes (Bayes factor > 6 of viral exchange among systems. Results indicate that currently circulating viruses are evolving rapidly, and show a higher level of relative genetic diversity over time, when compared to earlier relatives. Biological soundness of model results is supported by the finding that sow farms were responsible for PRRSV spread within the systems. Such results can’t be obtained by traditional phylogenetic methods, and therefore, our results provide a methodological framework for molecular epidemiological modeling of new PRRSV outbreaks and demonstrate the prospects of phylodynamic

  19. Virological and immunological responses to porcine reproductive and respiratory syndrome virus in a large population of gilts.

    Science.gov (United States)

    Batista, Laura; Pijoan, Carlos; Dee, Scott; Olin, Michael; Molitor, Thomas; Joo, Han Soo; Xiao, Zhenguo; Murtaugh, Michael

    2004-10-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes a prolonged active infection followed by a persistent infection in lymphoid tissues lasting for several months. Pigs develop both an antibody and cell-mediated immune response following PRRSV infection, but the specific role of each type in the development of protective immunity and clearance of the virus is not yet known. The aims of this study were to characterize the dynamics of PRRSV persistence from 0 to 135 d post infection (pi), characterize the kinetics of the antibody mediated immune response following PRRSV infection, and characterize the cell mediated immune responses to PRRSV infection. Eighty, 4-month-old PRRSV-free gilts were obtained from a source known to be negative for PRRSV. On day 0, gilts were infected intranasally with 10(2.4) TCID/50 MN 30-100 PRRSV. Following infection, animals were bled between days 0 to 135 pi. Viremia was detected up to day 30. Serum antibody response (by enzyme-linked immunosorbent assay [ELISA] and virus neutralization antibody) was detected from day 14 to 120 pi. Cell-mediated immune response represented by interferon gamma (IFN-gamma) was detected from day 14 to 120 pi. Persistence of PRRSV in tissues was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) between days 30 to 135. These results indicate that serum neutralizing antibodies and IFN-gamma play an important role in the clearance of PRRSV. Nevertheless none of the parameters measured (virus neutralizing antibodies), either alone or in combination, are solely responsible for the clearance of the virus from the host and the development of sterilizing immunity.

  20. Virological and immunological responses to porcine reproductive and respiratory syndrome virus in a large population of gilts

    Science.gov (United States)

    2004-01-01

    Abstract Porcine reproductive and respiratory syndrome virus (PRRSV) causes a prolonged active infection followed by a persistent infection in lymphoid tissues lasting for several months. Pigs develop both an antibody and cell-mediated immune response following PRRSV infection, but the specific role of each type in the development of protective immunity and clearance of the virus is not yet known. The aims of this study were to characterize the dynamics of PRRSV persistence from 0 to 135 d post infection (pi), characterize the kinetics of the antibody mediated immune response following PRRSV infection, and characterize the cell mediated immune responses to PRRSV infection. Eighty, 4-month-old PRRSV-free gilts were obtained from a source known to be negative for PRRSV. On day 0, gilts were infected intranasally with 102.4 TCID/ 50 MN 30–100 PRRSV. Following infection, animals were bled between days 0 to 135 pi. Viremia was detected up to day 30. Serum antibody response (by enzyme-linked immunosorbent assay [ELISA] and virus neutralization antibody) was detected from day 14 to 120 pi. Cell-mediated immune response represented by interferon gamma (IFN-γ) was detected from day 14 to 120 pi. Persistence of PRRSV in tissues was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) between days 30 to 135. These results indicate that serum neutralizing antibodies and IFN-γ play an important role in the clearance of PRRSV. Nevertheless none of the parameters measured (virus neutralizing antibodies), either alone or in combination, are solely responsible for the clearance of the virus from the host and the development of sterilizing immunity. PMID:15581221

  1. A molecular clock dates the common ancestor of European-type porcine reproductive and respiratory syndrome virus at more than 10 years before the emergence of disease

    DEFF Research Database (Denmark)

    Forsberg, Roald; Oleksiewicz, Martin B.; Krabbe Petersen, Anne Mette

    2001-01-01

    The disease caused by porcine reproductive and respiratory syndrome virus (PRRSV) emerged independently and almost simultaneously in Europe (1990) and North America (1987). The original reservoir of the virus and the date it entered the pig populations is not known. In this study, we demonstrate...... an accurate molecular clock for the European PRRSV ORF 3 gene, place the root in the genealogy, estimate the rate of nucleotide substitution, and date the most recent common viral ancestor of the data set to 1979; more than 10 years before the onset of the European epidemic. Based on these findings, we...

  2. Analysis of ORF 1 in European porcine reproductive and respiratory syndrome virus by long RT-PCR and restriction fragment length polymorphism (RFLP) analysis

    DEFF Research Database (Denmark)

    Nielsen, H. S.; Storgaard, Torben; Oleksiewicz, M.B.

    2000-01-01

    A rapid method was developed for partial characterization of the replicase-encoding open reading frame 1 (ORF 1) of porcine reproductive and respiratory syndrome virus (PRRSV). It comprised long RT-PCR amplification of 11.1 kb (94%) of ORF 1, followed by restriction fragment length polymorphism...... analysis. The method was used to compare ORF 1 sequences of two divergent European-type PRRSV strains. Our results indicated that the structural and replicase parts of these two strains had evolved at overall similar rates....

  3. Pathogenesis of porcine reproductive and respiratory syndrome: evaluation of the expression of cytokines and apoptosis phenomena in lymphoid organs and their role in the immune response

    OpenAIRE

    Barranco Cabezudo, Inmaculada

    2011-01-01

    El Síndrome Reproductivo y Respiratorio Porcino (PRRS, del inglés Porcine Reproductive and Respiratory Syndrome) es una enfermedad vírica caracterizada por inducir una respuesta inmune errática en el hospedador y es considerada como una de las enfermedades más importantes en la industria del porcino debido a las importantes pérdidas económicas que provoca. A pesar de que varios estudios se han realizado con el objetivo de elucidar la respuesta inmune provocada frente al virus del PRRS (PRRSV,...

  4. Examination of virus shedding in semen from vaccinated and from previously infected boars after experimental challenge with porcine reproductive and respiratory syndrome virus

    DEFF Research Database (Denmark)

    Nielsen, Thomas L.; Nielsen, Jens; Have, Per

    1997-01-01

    Danish artificial insemination (AI) centres house several boars antibody positive to porcine reproductive and respiratory syndrome virus as well as PRRSV-naive boars which may become acutely infected, The risk of transmission of PRRSV by semen may therefore constitute a serious problem...... vaccine compared to the non-vaccinated control animals. In contrast, no changes in onset, level and duration of viremia and shedding of virus in semen were observed using the inactivated vaccine, Neither viremia nor seminal shedding of virus was detected in previously PRRSV-infected, PRRSV...

  5. Determination of 5 '-leader sequences from radically disparate strains of porcine reproductive and respiratory syndrome virus reveals the presence of highly conserved sequence motifs

    DEFF Research Database (Denmark)

    Oleksiewicz, M.B.; Bøtner, Anette; Nielsen, Jens

    1999-01-01

    We determined the untranslated 5'-leader sequence for three different isolates of porcine reproductive and respiratory syndrome virus (PRRSV): pathogenic European- and American-types, as well as an American-type vaccine strain. 5'-leader from European- and American-type PRRSV differed in length...... (220 and 190 nt, respectively), and exhibited only approximately 50% nucleotide homology. Nevertheless, highly conserved areas were identified in the leader of all 3 PRRSV isolates, which constitute candidate motifs for binding of protein(s) involved in viral replication. These comparative data provide...

  6. Monitoring porcine reproductive and respiratory syndrome virus infection status in swine herds based on analysis of antibodies in meat juice samples

    DEFF Research Database (Denmark)

    Mortensen, Sten; Strandbygaard, Bertel; Bøtner, Anette

    2001-01-01

    An indirect ELISA test was developed as a novel tool aimed at monitoring the herd infection status of swine herds. Meat juice samples from pig carcasses were analysed for the presence of antibodies against porcine reproductive and respiratory syndrome virus (PRRSV). A study of samples from herds...... was validated in 47 herds by collection of blood samples from the herds. Eighteen herds were classified as PRRS negative by both test systems. Twenty-nine herds were classified as PRRS seropositive by both test systems. Acceptable herd classification was achieved using this test....

  7. Porcine reproductive and respiratory syndrome virus: Interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods

    DEFF Research Database (Denmark)

    Wernike, Kerstin; Bonilauri, Paolo; Dauber, Malte

    2012-01-01

    To compare the real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays used for the diagnosis of Porcine reproductive and respiratory syndrome virus (PRRSV), a Europe-wide interlaboratory ring trial was conducted. A variety of PRRSV strains including North American...... (NA) and European (EU) genotype isolates were analyzed by the participants. Great differences regarding qualitative diagnostics as well as analytical sensitivity were observed between the individual RT-qPCR systems, especially when investigating strains from the EU genotype. None of the assays...

  8. Effect of saliva stabilisers on detection of porcine reproductive and respiratory syndrome virus in oral fluid by quantitative reverse transcriptase real-time PCR.

    Science.gov (United States)

    Decorte, Inge; Van der Stede, Yves; Nauwynck, Hans; De Regge, Nick; Cay, Ann Brigitte

    2013-08-01

    This study evaluated the effect of extraction-amplification methods, storage temperature and saliva stabilisers on detection of porcine reproductive and respiratory syndrome virus (PRRSV) RNA by quantitative reverse transcriptase real-time PCR (qRT-PCR) in porcine oral fluid. The diagnostic performance of different extraction-amplification methods was examined using a dilution series of oral fluid spiked with PRRSV. To determine RNA stability, porcine oral fluid, with or without commercially available saliva stabilisers, was spiked with PRRSV, stored at 4°C or room temperature and tested for the presence of PRRSV RNA by qRT-PCR. PRRSV RNA could be detected in oral fluid using all extraction-amplification combinations, but the limit of detection varied amongst different combinations. Storage temperature and saliva stabilisers had an effect on the stability of PRRSV RNA, which could only be detected for 7 days when PRRSV spiked oral fluid was kept at 4°C or stabilised at room temperature with a commercial mRNA stabiliser. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Pig immune response to general stimulus and to porcine reproductive and respiratory syndrome virus infection: a meta-analysis approach.

    Science.gov (United States)

    Badaoui, Bouabid; Tuggle, Christopher K; Hu, Zhiliang; Reecy, James M; Ait-Ali, Tahar; Anselmo, Anna; Botti, Sara

    2013-04-03

    The availability of gene expression data that corresponds to pig immune response challenges provides compelling material for the understanding of the host immune system. Meta-analysis offers the opportunity to confirm and expand our knowledge by combining and studying at one time a vast set of independent studies creating large datasets with increased statistical power. In this study, we performed two meta-analyses of porcine transcriptomic data: i) scrutinized the global immune response to different challenges, and ii) determined the specific response to Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection. To gain an in-depth knowledge of the pig response to PRRSV infection, we used an original approach comparing and eliminating the common genes from both meta-analyses in order to identify genes and pathways specifically involved in the PRRSV immune response. The software Pointillist was used to cope with the highly disparate data, circumventing the biases generated by the specific responses linked to single studies. Next, we used the Ingenuity Pathways Analysis (IPA) software to survey the canonical pathways, biological functions and transcription factors found to be significantly involved in the pig immune response. We used 779 chips corresponding to 29 datasets for the pig global immune response and 279 chips obtained from 6 datasets for the pig response to PRRSV infection, respectively. The pig global immune response analysis showed interconnected canonical pathways involved in the regulation of translation and mitochondrial energy metabolism. Biological functions revealed in this meta-analysis were centred around translation regulation, which included protein synthesis, RNA-post transcriptional gene expression and cellular growth and proliferation. Furthermore, the oxidative phosphorylation and mitochondria dysfunctions, associated with stress signalling, were highly regulated. Transcription factors such as MYCN, MYC and NFE2L2 were found in

  10. Land altitude, slope, and coverage as risk factors for Porcine Reproductive and Respiratory Syndrome (PRRS outbreaks in the United States.

    Directory of Open Access Journals (Sweden)

    Andréia Gonçalves Arruda

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS is, arguably, the most impactful disease on the North American swine industry. The Swine Health Monitoring Project (SHMP is a national volunteer initiative aimed at monitoring incidence and, ultimately, supporting swine disease control, including PRRS. Data collected through the SHMP currently represents approximately 42% of the sow population of the United States. The objective of the study here was to investigate the association between geographical factors (including land elevation, and land coverage and PRRS incidence as recorded in the SHMP. Weekly PRRS status data from sites participating in the SHMP from 2009 to 2016 (n = 706 was assessed. Number of PRRS outbreaks, years of participation in the SHMP, and site location were collected from the SHMP database. Environmental features hypothesized to influence PRRS risk included land coverage (cultivated areas, shrubs and trees, land altitude (in meters above sea level and land slope (in degrees compared to surrounding areas. Other risk factors considered included region, production system to which the site belonged, herd size, and swine density in the area in which the site was located. Land-related variables and pig density were captured in raster format from a number of sources and extracted to points (farm locations. A mixed-effects Poisson regression model was built; and dependence among sites that belonged to a given production system was accounted for using a random effect at the system level. The annual mean and median number of outbreaks per farm was 1.38 (SD: 1.6, and 1 (IQR: 2.0, respectively. The maximum annual number of outbreaks per farm was 9, and approximately 40% of the farms did not report any outbreak. Results from the final multivariable model suggested that increments of swine density and herd size increased the risk for PRRS outbreaks (P < 0.01. Even though altitude (meters above sea level was not significant in the final

  11. Generation of porcine reproductive and respiratory syndrome (PRRS) virus-like-particles (VLPs) with different protein composition.

    Science.gov (United States)

    García Durán, Marga; Costa, Sofia; Sarraseca, Javier; de la Roja, Nuria; García, Julia; García, Isabel; Rodríguez, Maria José

    2016-10-01

    The causative agent of Porcine Reproductive and Respiratory Syndrome (PRRS) is an enveloped ssRNA (+) virus belonging to the Arteriviridae family. Gp5 and M proteins form disulfide-linked heterodimers that constitute the major components of PRRSV envelope. Gp2, Gp3, Gp4 and E are the minor structural proteins, being the first three incorporated as multimeric complexes in the virus surface. The disease has become one of the most important causes of economic losses in the swine industry. Despite efforts to design an effective vaccine, the available ones allow only partial protection. In the last years, VLPs have become good vaccine alternatives because of safety issues and their potential to activate both branches of the immunological response. The characteristics of recombinant baculoviruses as heterologous expression system have been exploited for the production of VLPs of a wide variety of viruses. In this work, two multiple baculovirus expression vectors (BEVs) with PRRS virus envelope proteins were engineered in order to generate PRRS VLPs: on the one hand, Gp5 and M cDNAs were cloned to generate the pBAC-Gp5M vector; on the other hand, Gp2, Gp3, Gp4 and E cDNAs have been cloned to generate the pBAC-Gp234E vector. The corresponding recombinant baculoviruses BAC-Gp5M and BAC-Gp234E were employed to produce two types of VLPs: basic Gp5M VLPs, by the simultaneous expression of Gp5 and M proteins; and complete VLPs, by the co-expression of the six PRRS proteins after co-infection. The characterization of VLPs by Western blot confirmed the presence of the recombinant proteins using the available specific antibodies (Abs). The analysis by Electron microscopy showed that the two types of VLPs were indistinguishable between them, being similar in shape and size to the native PRRS virus. This system represents a potential alternative for vaccine development and a useful tool to study the implication of specific PRRS proteins in the response against the virus. Copyright

  12. Induction of Apoptosis by the Nonstructural Protein 4 and 10 of Porcine Reproductive and Respiratory Syndrome Virus.

    Directory of Open Access Journals (Sweden)

    Shuaizhen Yuan

    Full Text Available Infection by most viruses triggers apoptosis in host cells, and viruses manipulate this cell response to promote viral replication, virus spread, and cell killing. Porcine reproductive and respiratory syndrome virus (PRRSV has been shown to induce apoptosis both in vitro and in vivo, while the regulatory roles of PRRSV-encoded products in apoptosis are not fully understood. In the present study, we first showed a biphasic apoptosis regulation by a highly pathogenic PRRSV strain JXwn06. It was indicated that PRRSV infection delays apoptosis at early infection but activates apoptosis at late infection in MARC-145 cells. In PRRSV-infected MARC-145 cells, procaspase-8, -9 and -12 were activated at late infection, demonstrating the involvements of death receptor pathway, mitochondrial pathway and endoplasmic reticulum (ER stress pathway in inducing apoptosis. PRRSV was also shown to induce a similar apoptosis process in pulmonary alveolar macrophages (PAMs with an early initiation. Next, the PRRSV-encoded apoptosis inducers were screened, indicating that the nonstructural protein (Nsp 4 and Nsp10 of PRRSV are pro-apoptotic. In the presence of Nsp4, it was confirmed that procaspase-8, -9 and -12 were cleaved, and Nsp4 facilitates the cleavage of procaspase-9 by activating B-cell lymphoma 2 interacting mediator of cell death (Bim, a pro-apoptotic protein. In addition, Nsp4 was shown to induce the degradation of an anti-apoptotic protein, B-cell lymphoma-extra large (Bcl-xL. Nsp10 was shown to activate procaspase-8 and -9 but procaspase-12 and to upregulate the expression of BH3-only pro-apoptotic protein BH3 interacting-domain death agonist (Bid and its active form, truncated Bid (tBid. Clearly, the participation of both activated caspase-8 and Bid is required for Nsp10-induced apoptosis, indicating a crosstalk between extrinsic- and mitochondria-dependent pathways. Together, our findings suggest that PRRSV infection regulates apoptosis in a two

  13. Induction of Apoptosis by the Nonstructural Protein 4 and 10 of Porcine Reproductive and Respiratory Syndrome Virus.

    Science.gov (United States)

    Yuan, Shuaizhen; Zhang, Ning; Xu, Lei; Zhou, Lei; Ge, Xinna; Guo, Xin; Yang, Hanchun

    2016-01-01

    Infection by most viruses triggers apoptosis in host cells, and viruses manipulate this cell response to promote viral replication, virus spread, and cell killing. Porcine reproductive and respiratory syndrome virus (PRRSV) has been shown to induce apoptosis both in vitro and in vivo, while the regulatory roles of PRRSV-encoded products in apoptosis are not fully understood. In the present study, we first showed a biphasic apoptosis regulation by a highly pathogenic PRRSV strain JXwn06. It was indicated that PRRSV infection delays apoptosis at early infection but activates apoptosis at late infection in MARC-145 cells. In PRRSV-infected MARC-145 cells, procaspase-8, -9 and -12 were activated at late infection, demonstrating the involvements of death receptor pathway, mitochondrial pathway and endoplasmic reticulum (ER) stress pathway in inducing apoptosis. PRRSV was also shown to induce a similar apoptosis process in pulmonary alveolar macrophages (PAMs) with an early initiation. Next, the PRRSV-encoded apoptosis inducers were screened, indicating that the nonstructural protein (Nsp) 4 and Nsp10 of PRRSV are pro-apoptotic. In the presence of Nsp4, it was confirmed that procaspase-8, -9 and -12 were cleaved, and Nsp4 facilitates the cleavage of procaspase-9 by activating B-cell lymphoma 2 interacting mediator of cell death (Bim), a pro-apoptotic protein. In addition, Nsp4 was shown to induce the degradation of an anti-apoptotic protein, B-cell lymphoma-extra large (Bcl-xL). Nsp10 was shown to activate procaspase-8 and -9 but procaspase-12 and to upregulate the expression of BH3-only pro-apoptotic protein BH3 interacting-domain death agonist (Bid) and its active form, truncated Bid (tBid). Clearly, the participation of both activated caspase-8 and Bid is required for Nsp10-induced apoptosis, indicating a crosstalk between extrinsic- and mitochondria-dependent pathways. Together, our findings suggest that PRRSV infection regulates apoptosis in a two-phase manner and

  14. Porcine reproductive and respiratory syndrome virus neutralizing antibodies provide in vivo cross-protection to PRRSV1 and PRRSV2 viral challenge.

    Science.gov (United States)

    Robinson, Sally R; Rahe, Michael C; Gray, Diem K; Martins, Kyra V; Murtaugh, Michael P

    2018-02-03

    Vaccine control and prevention of porcine reproductive and respiratory syndrome (PRRS), the most important disease of swine, is difficult to achieve. However, the discovery of broadly neutralizing antibody activity against porcine reproductive and respiratory syndrome virus (PRRSV) under typical field conditions opens the door to new immunologic approaches for robust protection. We show here that passive administration of purified immunoglobulins with neutralizing antibodies reduced PRRSV2 infection by up to 96%, and PRRSV1 infection by up to 87%, whereas immune immunoglobulins lacking neutralizing activity had no effect on viral infection. Hence, immune competence of passive immunoglobulin transfer was associated specifically with antibody neutralizing activity. Current models of PRRSV infection implicate a minor envelope glycoprotein (GP) complex including GP2, GP3, and GP4, as critical to permissive cell infection. However, conserved peptides comprising the putative cell attachment structure did not attenuate neutralization or viral infection. The results show that immunological approaches aimed at induction of broadly neutralizing antibodies may substantially enhance immune protection against PRRSV. The findings further show that naturally occurring viral isolates are able to induce protective humoral immunity against unrelated PRRSV challenge, thus removing a major conceptual barrier to vaccine development. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Porcine Reproductive and Respiratory Syndrome Virus Induces IL-1β Production Depending on TLR4/MyD88 Pathway and NLRP3 Inflammasome in Primary Porcine Alveolar Macrophages

    Directory of Open Access Journals (Sweden)

    Jing Bi

    2014-01-01

    Full Text Available Porcine reproductive and respiratory syndrome virus (PRRSV is an Arterivirus that has been devastating the swine industry worldwide since the late 1980s. Previous studies have reported that PRRSV infection induced the production of IL-1β. However, the cellular sensors and signaling pathways involved in this process have not been elucidated yet. Here, we studied the mechanisms responsible for the production of IL-1β in response to highly pathogenic PRRSV. Upon PRRSV infection of primary porcine alveolar macrophages, both mRNA expression and secretion of IL-1β were significantly increased in a time- and dose-dependent manner. We also investigated the role of several pattern-recognition receptors and adaptor molecules in this response and showed that the TLR4/MyD88 pathway and its downstream signaling molecules, NF-κB, ERK1/2, and p38 MAPKs, were involved in IL-1β production during PRRSV infection. Treatment with specific inhibitors or siRNA knockdown assays demonstrated that components of the NLRP3 inflammasome were crucial for IL-1β secretion but not for IL-1β mRNA expression. Furthermore, TLR4/MyD88/NF-κB signaling pathway was involved in PRRSV-induced expression of NLRP3 inflammasome components. Together, our results deciphered the pathways leading from recognition of PRRSV to the production and release of IL-1β, providing a deeper knowledge of the mechanisms of PRRSV-induced inflammation responses.

  16. Reproduction of postweaning multisystemic wasting syndrome in pigs by prenatal porcine circovirus 2 infection and postnatal porcine parvovirus infection or immunostimulation.

    Science.gov (United States)

    Ha, Y; Lee, Y H; Ahn, K-K; Kim, B; Chae, C

    2008-11-01

    Postweaning multisystemic wasting syndrome (PMWS) was reproduced in prenatally porcine circovirus 2 (PCV2)-infected pigs by either postnatal infection with porcine parvovirus (PPV) or by immunostimulation. Twenty-four randomly selected piglets from 3 sows, which had been experimentally infected during gestation with PCV2, were randomly divided into 3 groups; group 1 (prenatal PCV2 infection, with postnatal PPV infection), group 2 (prenatal PCV2 infection, with postnatal keyhole limpet hemocyanin, emulsified in incomplete Freund's adjuvant [KLH/ICFA] injection), and group 3 (prenatal PCV2 infection only). Twenty-four randomly selected piglets from 3 uninfected sows were randomly divided into 3 groups; group 4 (no prenatal infection, with postnatal PCV2 and PPV infection), group 5 (no prenatal infection, with postnatal PCV2 infection), and group 6 (negative control pigs). Body weight in negative control pigs (group 6) was increased significantly compared with pigs in groups 1, 2, and 4 at 49, 52, 56, 59, and 63 days of age. The granulomatous inflammatory reaction and lymphoid depletion that are typical lesions in pigs with PMWS were observed in the lymph node of piglets in groups 1, 2, and 4 at 63 days of age. Pigs in group 3 had significantly fewer PCV2-positive cells than those from groups 1, 2, 4, or 5. When the prenatally PCV2-infected pigs were infected with PPV or injected with immunostimulant in the postnatal period, they developed PMWS. Thus, factors that potentiate the progression of prenatal PCV2 infection to PMWS are postnatal infection with PPV or immune stimulation.

  17. Highly efficient expression of interleukin-2 under the control of rabbit β-globin intron II gene enhances protective immune responses of porcine reproductive and respiratory syndrome (PRRS DNA vaccine in pigs.

    Directory of Open Access Journals (Sweden)

    Yijun Du

    Full Text Available Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV had caused catastrophic losses in swine industry in China. The current inactivated vaccine provided only limited protection, and the attenuated live vaccine could protect piglets against the HP-PRRSV but there was a possibility that the attenuated virus returned to high virulence. In this study, the eukaryotic expression vector pVAX1© was modified under the control of rabbit β-globin intron II gene and the modified vector pMVAX1© was constructed. Porcine interleukin-2 (IL-2 and GP3-GP5 fusion protein of HP-PRRSV strain SD-JN were highly expressed by pMVAX1©. Mice inoculated with pMVAX1©-GP35 developed significantly higher PRRSV-specific antibody responses and T cell proliferation than those vaccinated with pVAX1©-GP35. pMVAX1©-GP35 was selected as PRRS DNA vaccine candidate and co-administrated with pVAX1©-IL-2 or pMVAX1©-IL-2 in pigs. pMVAX1©-IL-2+pMVAX1©-GP35 could provide enhanced PRRSV-specific antibody responses, T cell proliferation, Th1-type and Th2-type cytokine responses and CTL responses than pMVAX1©-GP35 and pVAX1©-IL-2+pMVAX1©-GP35. Following homologous challenge with HP-PRRSV strain SD-JN, similar with attenuated PRRS vaccine group, pigs inoculated with pMVAX1©-IL-2+pMVAX1©-GP35 showed no clinical signs, almost no lung lesions and no viremia, as compared to those in pMVAX1©-GP35 and pVAX1©-IL-2+pMVAX1©-GP35 groups. It indicated that pMVAX1©-IL-2 effectively increases humoral and cell mediated immune responses of pMVAX1©-GP35. Co-administration of pMVAX1©-IL-2 and pMVAX1©-GP35 might be attractive candidate vaccines for preventing HP-PRRSV infections.

  18. Analysis of ORF5 and Full-Length Genome Sequences of Porcine Reproductive and Respiratory Syndrome Virus Isolates of Genotypes 1 and 2 Retrieved Worldwide Provides Evidence that Recombination Is a Common Phenomenon and May Produce Mosaic Isolates

    DEFF Research Database (Denmark)

    Martín-Valls, G. E.; Kvisgaard, Lise Kirstine; Tello, M.

    2014-01-01

    Recombination is currently recognized as a factor for high genetic diversity, but the frequency of such recombination events and the genome segments involved are not well known. In the present study, we initially focused on the detection of recombinant porcine reproductive and respiratory syndrom...

  19. Development of a swine specific 9-plex Luminex cytokine assay and assessment of immunity after porcine reproductive and respiratory syndrome virus (PRRSV) vaccination: Elevated serum IL-12 levels are not predictive of protect

    Science.gov (United States)

    A Luminex multiplex swine cytokine assay was developed to measure 9 cytokines simultaneously in pig serum and tested in a porcine reproductive and respiratory syndrome virus (PRRSV) vaccine/challenge study. This assay detects innate (IL-1ß, IL-6, IL-8, IFNa, TNFa); regulatory (IL-10), Th1 (IL-12, I...

  20. Preparation for emergence of an Eastern European porcine reproductive and respiratory syndrome virus (PRRSV) strain in Western Europe: Immunization with modified live virus vaccines or a field strain confers partial protection.

    Science.gov (United States)

    Renson, P; Fablet, C; Le Dimna, M; Mahé, S; Touzain, F; Blanchard, Y; Paboeuf, F; Rose, N; Bourry, O

    2017-05-01

    The porcine reproductive and respiratory syndrome virus (PRRSV) causes huge economic losses for the swine industry worldwide. In the past several years, highly pathogenic strains that lead to even greater losses have emerged. For the Western European swine industry, one threat is the possible introduction of Eastern European PRRSV strains (example Lena genotype 1.3) which were shown to be more virulent than common Western resident strains under experimental conditions. To prepare for the possible emergence of this strain in Western Europe, we immunized piglets with a Western European PRRSV field strain (Finistere: Fini, genotype 1.1), a new genotype 1 commercial modified live virus (MLV) vaccine (MLV1) or a genotype 2 commercial MLV vaccine (MLV2) to evaluate and compare the level of protection that these strains conferred upon challenge with the Lena strain 4 weeks later. Results show that immunization with Fini, MLV1 or MLV2 strains shortened the Lena-induced hyperthermia. In the Fini group, a positive effect was also demonstrated in growth performance. The level of Lena viremia was reduced for all immunized groups (significantly so for Fini and MLV2). This reduction in Lena viremia was correlated with the level of Lena-specific IFNγ-secreting cells. In conclusion, we showed that a commercial MLV vaccine of genotype 1 or 2, as well as a field strain of genotype 1.1 may provide partial clinical and virological protection upon challenge with the Lena strain. The cross-protection induced by these immunizing strains was not related with the level of genetic similarity to the Lena strain. The slightly higher level of protection established with the field strain is attributed to a better cell-mediated immune response. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. A novel recombinant virus-like particle vaccine for prevention of porcine parvovirus-induced reproductive failure

    NARCIS (Netherlands)

    Antonis, A.F.G.; Bruschke, C.J.M.; Rueda, P.; Maranga, L.; Casal, J.; Vela, C.; Hilgers, L.A.T.; Belt, P.B.G.M.; Weerdmeester, K.; Carrondo, M.J.; Langeveld, J.P.M.

    2006-01-01

    A novel vaccine against porcine parvovirus (PPV), composed of recombinant virus-like particles (PPV-VLPs) produced with the baculovirus expression vector system (BEVS) at industrial scale, was tested for its immunogenicity and protective potency. A formulation of submicrogram amounts of PPV-VLPs in

  2. Influence of the amino acid residues at 70 in M protein of porcine reproductive and respiratory syndrome virus on viral neutralization susceptibility to the serum antibody.

    Science.gov (United States)

    Fan, Baochao; Liu, Xing; Bai, Juan; Zhang, Tingjie; Zhang, Qiaoya; Jiang, Ping

    2016-03-22

    Porcine reproductive and respiratory syndrome virus (PRRSV) is mainly responsible for the significant economic losses in pig industry in the world. The adaptive immune responses of the host act as an important source of selective pressure in the evolutionary process of the virus. In the previous study, we confirmed that the amino acid (aa) residues at 102 and 104 sites in GP5 played an important role in escaping from the neutralizing antibodies (NAbs) against highly pathogenic PRRSV (HP-PRRSV). In this study, we further analyzed the aa mutants affecting neutralization susceptibility of NAbs in other structure proteins in NAbs resistant variants. Based on the different aa residues of the structural proteins between the resistant virus BB20s and the parent virus BB, 12 recombinant PRRSV strains containing these aa residue substitutions were constructed using reverse genetic techniques. The neutralizing antibody (NA) titers of the recombinant strains were tested on MARC-145 and porcine alveolar macrophages (PAMs). And the NAbs binding abilities of parent and rescued viruses were tested by using ELISA method. By using the neutralization assay, it was revealed that the NA titer of N4 serum with rBB/Ms was significantly lower than that with rBB. Meanwhile, NA titer of the serum with rBB20s/M was significantly higher than that with rBB20s. The ELISA binding results showed that rBB/Ms had higher binding inability to N4 than did rBB. And alignment of M protein revealed that the variant aa residue lysine (K) at 70 was also existed in field type 2 and vaccine PRRSV strains. The aa residue at 70 in M protein of PRRSV played an important role in regulating neutralization susceptibility to the porcine serum NAbs. It may be helpful for monitoring the antigen variant strains in the field and developing new vaccine against PRRSV in the future.

  3. Poly(I:C) inhibits porcine reproductive and respiratory syndrome virus replication in MARC-145 cells via activation of IFIT3.

    Science.gov (United States)

    Zhang, Lili; Liu, Jie; Bai, Juan; Du, Yijun; Wang, Xiaoye; Liu, Xing; Jiang, Ping

    2013-09-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major cause of heavy economic losses in many swine-producing regions. Current vaccination strategies and antiviral drugs provide only limited protection. Interferon (IFN)-induced protein with tetratricopeptide repeats 3 (IFIT3) has been characterized as the product of a novel antiviral gene and as an important modulator in innate immunity. However, the role of IFIT3 in PRRSV infection is scarcely understood. In this study, polyinosinic-polycytidylic acid (poly(I:C)) inhibited PRRSV replication in MARC-145 cells, following the appearance of increased IFIT3. Overexpression of porcine IFIT3 resulted in a decrease of PRRSV. Knockdown of IFIT3 in MARC-145 cells increased PRRSV replication and impaired the antiviral activity mediated by poly(I:C). Moreover, in the presence or absence of IFIT3, poly(I:C)-induced IFN-β promoter activity was significantly boosted or crippled, respectively. IFIT3, TBK1 and phosphorylation of IRF3 were activated in poly(I:C)-transfected MARC-145 cells. It demonstrated that IFIT3 plays an important role in IFN-β induction in MARC-145 cells, and, when activated, it can inhibit PRRSV replication. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Interleukin-10 antisense oligodeoxynucleotide suppresses IL-10 expression and effects on proinflammatory cytokine responses to porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Charerntantanakul, Wasin; Kasinrerk, Watchara

    2010-08-01

    Upregulation of interleukin-10 (IL-10) expression has been suggested to be the mechanism by which the porcine reproductive and respiratory syndrome virus (PRRSV) suppresses the innate and adaptive immune response in infected pigs. In this study we evaluated the potential of phosphorothioate-modified IL-10 antisense oligodeoxynucleotide specific to the translation initiation region of porcine IL-10 mRNA (IL-10AS) in enhancing proinflammatory cytokine responses to PRRSV. Naïve peripheral blood mononuclear cells from eight PRRSV-seronegative pigs were transfected with IL-10AS in vitro prior to PRRSV inoculation and phorbol 12-myristate 13-acetate plus ionomycin or concanavalin A stimulation. The effects of IL-10AS on mRNA expression of IL-10, interferon-gamma (IFN-gamma), IFN-alpha, tumor necrosis factor-alpha (TNF-alpha), IL-2, and IL-4 were tested by real-time PCR. The percentages of IFN-gamma-producing T-cell subsets were determined by flow cytometry. Compared to the controls, the levels of IL-10 and IL-2 mRNA were significantly reduced, while those of IFN-gamma mRNA were increased, and TNF-alpha, IFN-alpha, and IL-4 mRNA were unchanged. An increase in the percentage of the IFN-gamma+ population was also observed in lymphocytes and CD8beta+ T cells. Our results suggest that IL-10AS has the potential to enhance proinflammatory cytokine responses to PRRSV infection.

  5. Prevalence of porcine circovirus-2 DNA-positive ovarian and uterine tissues in gilts culled due to reproductive disturbance in Thailand.

    Science.gov (United States)

    Pearodwong, Pachara; Srisuwatanasagul, Sayamon; Teankum, Komkrich; Tantilertcharoen, Rachod; Tummaruk, Padet

    2015-06-01

    The present study aimed to determine the prevalence of porcine circovirus-2 (PCV-2) DNA-positive ovarian and uterine tissues in gilts culled due to reproductive disturbance in Thailand. Tissues (70 ovaries and 102 uteri) and serum (n = 102) samples from 102 gilts were included. PCV-2 DNA was detected by using polymerase chain reactions. The localisation of PCV-2 antigen was determined by immunohistochemistry, and PCV-2 antibody was evaluated by ELISA. PCV-2 DNA was detected in 30.0 % (21/70) of the ovaries and in 45.1 % (46/102) of the uteri. Age did not influence the frequency of PCV-2 DNA detection in these reproductive organs of gilts (P > 0.05). The prevalence of PCV-2 DNA-positive uterine tissue in gilts culled due to non-reproductive problems (20.0 %) was lower than gilts culled due to abortion (85.0 %), abnormal vaginal discharge (47.5 %) and anoestrus (53.5 %) (P gilts with high antibody titres (23.0 %) was lower than in gilts with low antibody titres (57.6 %) and seronegative gilts (64.5 %) (P gilts.

  6. Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No 2016/429): porcine reproductive and respiratory syndrome (PRRS)

    DEFF Research Database (Denmark)

    EFSA Panel on Animal Health and Welfare (AHAW); More, Simon J.; Bøtner, Anette

    2017-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on the eligibility of PRRS to be listed, Article 9 for the categorisation of PRRS according...... to disease prevention and control rules as in Annex IV and Article 8 on the list of animal species related to PRRS. The assessment has been performed following a methodology composed of information collection and compilation, expert judgement on each criterion at individual and, if no consensus was reached......) of Article 9(1). The animal species to be listed for PRRS according to Article 8(3) criteria are domestic pigs and wild boar....

  7. Full-length genome sequence analysis of a Hungarian porcine reproductive and respiratory syndrome virus isolated from a pig with severe respiratory disease.

    Science.gov (United States)

    Bálint, Ádám; Balka, Gyula; Horváth, Péter; Kecskeméti, Sándor; Dán, Ádám; Farsang, Attila; Szeredi, Levente; Bányai, Krisztián; Bartha, Dániel; Olasz, Ferenc; Belák, Sándor; Zádori, Zoltán

    2015-02-01

    Here, we report the isolation of a type 1 porcine reproductive and respiratory syndrome virus (PRRSV) strain from a clinical outbreak of severe respiratory problems and high fever. Next-generation sequencing was used to determine the complete genome sequence of the isolate (9625/2012). The virus belongs to a new branch within subtype 1, clade D, and shows the highest similarity to PRRSV Olot/1991 and to the Amervac vaccine strain. Mutation analysis of 9625/2012 revealed no evidence of recombination but did show a high proportion of amino acid substitutions in the putative neutralizing epitopes, suggesting an important role of selective immune pressure in the evolution of PRRSV 9625/2012.

  8. Genetic and antigenic drift of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in a closed population evaluated by full genome sequencing

    DEFF Research Database (Denmark)

    Kvisgaard, Lise Kirstine; Hjulsager, Charlotte Kristiane; Larsen, Lars Erik

    of circulating PRRS viruses in Danish pigs and to investigate the genetic drift of the virus in a closed population with very limited introductions of new animals. The study included phylogenetic analysis of full genome sequences of eight Type 1 and nine Type 2 PRRS viruses, including the very first Danish......Porcine Reproductive and Respiratory Syndrome (PRRS) viruses are divided into two major genotypes (Type 1 and Type 2) based on their genetic diversity. Type 1 PRRSV is further divided into at least 3 subtypes, but until now only subtype 1 has been detected in Western Europe and North America. Both...... isolated Type 1 virus and the very first Danish Type 2 PRRS virus isolated from a non-vaccinated pig herd. Furthermore, by sequencing ORF5 and ORF7 of 43 Type 1 and 57 Type 2 viruses isolated between 2003 and 2013, the level of genetic diversity was assessed. The results showed a very high genetic...

  9. In utero infection with porcine reproductive and respiratory syndrome virus modulates leukocyte subpopulations in peripheral blood and bronchoalveolar fluid of suviving piglets

    DEFF Research Database (Denmark)

    Nielsen, J.; Bøtner, Anette; Tingstedt, J. E.

    2003-01-01

    It is well known that piglets congenitally infected with porcine reproductive and respiratory syndrome virus (PRRSV) can be viremic at birth, and that preweaning mortality due to secondary infections often increases during acute outbreaks of PRRS. Therefore, an immunosuppressive effect of in utero...... infection has been suggested. The aim of the present study was to characterise the changes of leukocyte populations in piglets surviving in utero infection with PRRSV. A total of 27 liveborn uninfected control piglets and 22 piglets infected transplacentally with a Danish strain of PRRSV were included. At 2...... and 4 weeks of age, 21 of 22 (96%) and 7 of 14 (50%) examined infected piglets were still viremic, whereas PRRSV could not be detected in the six infected piglets examined at 6 weeks of age. Flow cytometry analysis was used to determine the phenotypic composition of leukocytes in peripheral blood...

  10. Modulation of Proinflammatory Cytokines in Monocyte-Derived Dendritic Cells by Porcine Reproductive and Respiratory Syndrome Virus Through Interaction with the Porcine Intercellular-Adhesion-Molecule-3-Grabbing Nonintegrin.

    Science.gov (United States)

    Piñeyro, Pablo E; Subramaniam, Sakthivel; Kenney, Scott P; Heffron, C Lynn; Giménez-Lirola, Luis G; Meng, Xiang-Jin

    2016-12-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important global swine pathogen. PRRSV infects porcine dendritic cells (DCs), but the effects of the interactions with DCs are largely unknown. Current research focuses on the production and regulation of interferons and selected inflammatory cytokines in DCs, which may play key roles in immune modulation. In addition, PRRSV also downregulates swine leukocyte antigen class I (SLA-I), SLA-II, and CD80/86 costimulatory molecules in DCs. In this study, we aim to evaluate the PRRSV immunomodulatory effects on monocyte-derived DCs (MoDCs) through interactions with porcine DC-SIGN (pDC-SIGN) receptor. We demonstrated that blocking the PRRSV and pDC-SIGN interactions in MoDCs with recombinant hICAM-3 did not affect the regulatory effects of PRRSV on SLA-I, SLA-II, or CD80/86 molecules. The hICAM-3 did not affect the morphological changes on MoDCs associated with their activation and maturation after PRRSV infection, and did not impair the virus infectivity in these cells either. The mRNA levels of tumor necrosis factor alpha (TNF-α), IL-12p35, IL-1β, and IL-6 were upregulated after hICAM-3 treatment or PRRSV infection, but in the presence of the blockage of pDC-SIGN in MoDCs with hICAM-3, PRRSV did not modulate the expression of these genes. However, in the presence of an anti-pDC-SIGN monoclonal antibody (mAb), we showed that PRRSV infection significantly reduced the mRNA expression levels of TNF-α and IL-1α, but enhanced the expression of IL-12p35 in MoDCs. Both hICAM-3-Fc and pDC-SIGN mAb treatments did not modulate proinflammatory cytokine protein levels in the culture supernatants of PRRSV-infected MoDCs. The results indicate that blocking the PRRSV-pDC-SIGN interactions by recombinant hICAM-3-Fc did not significantly affect virus infectivity, DC maturation, and proinflammatory cytokine gene expression in infected MoDCs. However, blocking the PRRSV-pDC-SIGN interactions on MoDCs with

  11. Long-term administration of a commercial porcine reproductive and respiratory syndrome virus (PRRSV)-inactivated vaccine in PRRSV-endemically infected sows.

    Science.gov (United States)

    Papatsiros, V G; Alexopoulos, C; Kritas, S K; Koptopoulos, G; Nauwynck, H J; Pensaert, M B; Kyriakis, S C

    2006-08-01

    The purpose of this study was to investigate the safety and efficacy of a commercial European porcine reproductive and respiratory syndrome virus (PRRSV)-inactivated vaccine after 18-month use in gilts/sows at a farm with high seroprevalence. In a farrow-to-finish farm with 1100 sows, all sows and gilts were systematically vaccinated with the PRRS-inactivated PROGRESSIS vaccine for a period of 18 months. Farm's reproductive and litter characteristics were longitudinally recorded for this period and historically compared with those of the year prior to vaccination. Serology, employing immunoperoxidase monolayer assay, had confirmed a high prevalence of PRRS-specific antibodies in most age groups within the farm prior to vaccination. Seroprevalence during the experiment ranged between 0% and 100% in weaners and growers, but remained at stable high levels (> 93%) in finishing pigs and gilts throughout all 2-year period of serology measurements. No local or systemic vaccine side effects were noted throughout the trial period. Vaccinations had resulted over time in a significant improvement of sow reproductive performance (e.g. reduction of premature farrowings, abortions and increase of farrowing rate) and litter characteristics (e.g. increase of the number of live born and weaned pigs and decrease of stillborn, mummified, weak and splay-legged piglets). It has also been observed that the higher the degree of immunization of a sow, the better the improvement of her reproductive parameters. Sows after vaccination have shown improved characteristics compared to homoparous sows prior to the application of vaccinations in the farm.

  12. Detection of genotype 1 Porcine Reproductive and Respiratory Syndrome virus in swine, using one-step Real-Time PCR for the ORF7 gene

    Directory of Open Access Journals (Sweden)

    Mihaela Zaulet

    2014-10-01

    Full Text Available Porcine Reproductive and Respiratory Syndrome (PRRS is the most devastating and economically challenging disease to the swine industry worldwide due to reproductive failure. The main objective of the current study was to evaluate the sensitivity and accuracy of Real-Time RT-PCR method in the detection of PRRS virus and also estimation of the pathogen load in samples with clinical signs. The primers used for the detection of PRRS virus were represented by primers with a specific sequence for the ORF7 gene of the PRRS virus. More important, the primers attachment process was influenced by punctual mutations of the viral strand belonging to the ORF7 gene. 114 samples were tested to identify the presence of PRRS virus, genotype I and 14 of them were found to be positive, using OneStep PCR. Those samples were used to test the specificity of the TaqMan probe and robustness of Real-Time RT-PCR reaction. According to the results, only the samples which presented some specific punctual mutations (4 in total, all from one particular region of Romania at the genome level of ORF7, were positive, due to primer sequence specificity and complementarity. The Real-Time RT-PCR method has been increasingly adopted by swine producers and veterinarian laboratories as one of the most trustful techniques, combining rapidity, specificity and efficiency for detecting and monitoring the spread of PRRS virus.

  13. Cross-protection of a new type 2 porcine reproductive and respiratory syndrome virus (PRRSV) modified live vaccine (Fostera PRRS) against heterologous type 1 PRRSV challenge in growing pigs.

    Science.gov (United States)

    Park, Changhoon; Choi, Kyuhyung; Jeong, Jiwoon; Chae, Chanhee

    2015-05-15

    The objective of the present study was to determine the cross-protection of a new type 2 porcine reproductive and respiratory syndrome virus (PRRSV) modified live vaccine against heterologous type 1 PRRSV challenge in growing pigs. The mean rectal temperature and respiratory score was significantly (Pprotection of a new type 2 PRRSV modified live vaccine against heterologous type 1 PRRSV challenge in growing pigs. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Transcriptome profile of lung dendritic cells after in vitro porcine reproductive and respiratory syndrome virus (PRRSV) infection

    DEFF Research Database (Denmark)

    Pröll, Maren Julia; Neuhoff, Christiane; Schellander, Karl

    2017-01-01

    generally defends against infectious diseases, especially dendritic cells (DCs), which play a crucial role in the activation of the immune response after viral infections. However, the understanding of the immune response and the genetic impact on the immune response to PRRS virus (PRRSV) remains incomplete....... In light of this, we investigated the regulation of the host immune response to PRRSV in porcine lung DCs using RNA-sequencing (RNA-Seq). Lung DCs from two different pig breeds (Pietrain and Duroc) were collected before (0 hours) and during various periods of infection (3, 6, 9, 12, and 24 hours post...... could be useful in investigations on immunity traits in pig breeding and enhancing the health of pigs....

  15. Genome-wide gene expression profiles in lung tissues of pig breeds differing in resistance to porcine reproductive and respiratory syndrome virus.

    Directory of Open Access Journals (Sweden)

    Jinyi Xing

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS caused by PRRS virus (PRRSV is an infectious disease characterized by severe reproductive deficiency in pregnant sows, typical respiratory symptoms in piglets, and high mortality rate of piglets. In this study, we employed an Affymetrix microarray chip to compare the gene expression profiles of lung tissue samples from Dapulian (DPL pigs (a Chinese indigenous pig breed and Duroc×Landrace×Yorkshire (DLY pigs after infection with PRRSV. During infection with PRRSV, the DLY pigs exhibited a range of clinical features that typify the disease, whereas the DPL pigs showed only mild signs of the disease. Overall, the DPL group had a lower percentage of CD4(+ cells and lower CD4(+/CD8(+ratios than the DLY group (p<0.05. For both IL-10 and TNF-α, the DLY pigs had significantly higher levels than the DPL pigs (p<0.01. The DLY pigs have lower serum IFN-γ levels than the DPL pigs (p<0.01. The serum IgG levels increased slightly from 0 dpi to 7 dpi, and peaked at 14 dpi (p<0.0001. Microarray data analysis revealed 16 differentially expressed (DE genes in the lung tissue samples from the DLY and DPL pigs (q≤5%, of which LOC100516029 and LOC100523005 were up-regulated in the PRRSV-infected DPL pigs, while the other 14 genes were down-regulated in the PRRSV-infected DPL pigs compared with the PRRSV-infected DLY pigs. The mRNA expression levels of 10 out of the 16 DE genes were validated by real-time quantitative RT-PCR and their fold change was consistent with the result of microarray data analysis. We further analyzed the mRNA expression level of 8 differentially expressed genes between the DPL and DLY pigs for both uninfected and infected groups, and found that TF and USP18 genes were important in underlying porcine resistance or susceptibility to PRRSV.

  16. Differences in Whole Blood Gene Expression Associated with Infection Time-Course and Extent of Fetal Mortality in a Reproductive Model of Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Infection

    Science.gov (United States)

    Wilkinson, Jamie M.; Ladinig, Andrea; Bao, Hua; Kommadath, Arun; Stothard, Paul; Lunney, Joan K.; Harding, John C. S.; Plastow, Graham S.

    2016-01-01

    Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection of pregnant females causes fetal death and increased piglet mortality, but there is substantial variation in the extent of reproductive pathology between individual dams. This study used RNA-sequencing to characterize the whole blood transcriptional response to type 2 PRRSV in pregnant gilts during the first week of infection (at 0, 2, and 6 days post-inoculation), and attempted to identify gene expression signatures associated with a low or high level of fetal mortality rates (LFM and HFM; n = 8/group) at necropsy, 21 days post-inoculation. The initial response to infection measured at 2 days post-inoculation saw an upregulation of genes involved in innate immunity, such as interferon-stimulated antiviral genes and inflammatory markers, and apoptosis. A concomitant decrease in expression of protein synthesis and T lymphocyte markers was observed. By day 6 the pattern had reversed, with a drop in innate immune signaling and an increase in the expression of genes involved in cell division and T cell signaling. Differentially expressed genes (DEGs) associated with extremes of litter mortality rate were identified at all three time-points. Among the 15 DEGs upregulated in LFM gilts on all three days were several genes involved in platelet function, including integrins ITGA2B and ITGB3, and the chemokine PF4 (CXCL4). LFM gilts exhibited a higher baseline expression of interferon-stimulated and pro-inflammatory genes prior to infection, and of T cell markers two days post-infection, indicative of a more rapid progression of the immune response to PRRSV. This study has increased our knowledge of the early response to PRRSV in the blood of pregnant gilts, and could ultimately lead to the development of a biomarker panel that can be used to predict PRRSV-associated reproductive pathology. PMID:27093427

  17. Differences in Whole Blood Gene Expression Associated with Infection Time-Course and Extent of Fetal Mortality in a Reproductive Model of Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV Infection.

    Directory of Open Access Journals (Sweden)

    Jamie M Wilkinson

    Full Text Available Porcine Reproductive and Respiratory Syndrome Virus (PRRSV infection of pregnant females causes fetal death and increased piglet mortality, but there is substantial variation in the extent of reproductive pathology between individual dams. This study used RNA-sequencing to characterize the whole blood transcriptional response to type 2 PRRSV in pregnant gilts during the first week of infection (at 0, 2, and 6 days post-inoculation, and attempted to identify gene expression signatures associated with a low or high level of fetal mortality rates (LFM and HFM; n = 8/group at necropsy, 21 days post-inoculation. The initial response to infection measured at 2 days post-inoculation saw an upregulation of genes involved in innate immunity, such as interferon-stimulated antiviral genes and inflammatory markers, and apoptosis. A concomitant decrease in expression of protein synthesis and T lymphocyte markers was observed. By day 6 the pattern had reversed, with a drop in innate immune signaling and an increase in the expression of genes involved in cell division and T cell signaling. Differentially expressed genes (DEGs associated with extremes of litter mortality rate were identified at all three time-points. Among the 15 DEGs upregulated in LFM gilts on all three days were several genes involved in platelet function, including integrins ITGA2B and ITGB3, and the chemokine PF4 (CXCL4. LFM gilts exhibited a higher baseline expression of interferon-stimulated and pro-inflammatory genes prior to infection, and of T cell markers two days post-infection, indicative of a more rapid progression of the immune response to PRRSV. This study has increased our knowledge of the early response to PRRSV in the blood of pregnant gilts, and could ultimately lead to the development of a biomarker panel that can be used to predict PRRSV-associated reproductive pathology.

  18. Differences in Whole Blood Gene Expression Associated with Infection Time-Course and Extent of Fetal Mortality in a Reproductive Model of Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Infection.

    Science.gov (United States)

    Wilkinson, Jamie M; Ladinig, Andrea; Bao, Hua; Kommadath, Arun; Stothard, Paul; Lunney, Joan K; Harding, John C S; Plastow, Graham S

    2016-01-01

    Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection of pregnant females causes fetal death and increased piglet mortality, but there is substantial variation in the extent of reproductive pathology between individual dams. This study used RNA-sequencing to characterize the whole blood transcriptional response to type 2 PRRSV in pregnant gilts during the first week of infection (at 0, 2, and 6 days post-inoculation), and attempted to identify gene expression signatures associated with a low or high level of fetal mortality rates (LFM and HFM; n = 8/group) at necropsy, 21 days post-inoculation. The initial response to infection measured at 2 days post-inoculation saw an upregulation of genes involved in innate immunity, such as interferon-stimulated antiviral genes and inflammatory markers, and apoptosis. A concomitant decrease in expression of protein synthesis and T lymphocyte markers was observed. By day 6 the pattern had reversed, with a drop in innate immune signaling and an increase in the expression of genes involved in cell division and T cell signaling. Differentially expressed genes (DEGs) associated with extremes of litter mortality rate were identified at all three time-points. Among the 15 DEGs upregulated in LFM gilts on all three days were several genes involved in platelet function, including integrins ITGA2B and ITGB3, and the chemokine PF4 (CXCL4). LFM gilts exhibited a higher baseline expression of interferon-stimulated and pro-inflammatory genes prior to infection, and of T cell markers two days post-infection, indicative of a more rapid progression of the immune response to PRRSV. This study has increased our knowledge of the early response to PRRSV in the blood of pregnant gilts, and could ultimately lead to the development of a biomarker panel that can be used to predict PRRSV-associated reproductive pathology.

  19. Síndrome reprodutiva e respiratória dos suínos: uma breve revisão Porcine reproductive and respiratory syndrome: a brief review

    Directory of Open Access Journals (Sweden)

    Luiz Carlos Kreutz

    1998-03-01

    Full Text Available A síndrome reprodutiva e respiratória dos suínos (Porcine Reproductive and Respiratory Syndrome - PRRS é uma doença relativamente nova dos suínos que foi detectada primeiramente em 1985 nos Estados Unidos, e em 1990 no continente Europeu. A síndrome é causada pelo PRRS vírus (PRRSV, o qual foi incluído em uma nova família de vírus, a Arteriviridae. A infecção pelo PRRSV causa problemas reprodutivos em fêmeas gestantes, o quais são caracterizados por abortos no final da gestação e/ou parto precoce, onde pode-se observar um elevado numero de fetos mumificados e natimortos; leitões que nascem infectados são fracos e economicamente inviáveis. Os problemas respiratórios causados pela infecção pelo PRRSV podem se manifestar em suínos de todas as faixas etárias, e são semelhantes a influenza. Embora PRRS tem sido detectada na maioria dos países em que a suinocultura tem importância econômica significativa, não há informações publicadas a respeito da doença ou do vírus no Brasil. No entanto, devido as perdas econômicas significativas que essa síndrome causou nos países já afetados, e da possibilidade do vírus ser eventualmente introduzido nos rebanhos brasileiros, é necessário reconhecer a doença imediatamente, e tomar as devidas medidas para o diagnóstico e controle em casos de surtos de problemas reprodutivos e respiratórios.Porcine reproductive and respiratory syndrome (PRRS is a relatively new disease of swine that emerged in the United States in the late 1980s and in Europe in 1990. The syndrome is caused by a virus, the PRRS virus (PRRSV which has been included into a newly proposed family of viruses, the Arteriviridae. Infection by PRRSV causes reproductive failure in pregnant females, characterized by late term abortion and early farowing, and an increased number of mummified and stillborn fetuses; newborn infected piglets are usually weak and unthrifty. Respiratory distress caused by PRRSV infection

  20. Porcine reproductive and respiratory syndrome virus (PRRSV) up-regulates IL-15 through PKCβ1-TAK1-NF-κB signaling pathway.

    Science.gov (United States)

    Du, Li; Liu, Yihao; Du, Yinping; Wang, Honglei; Zhang, Meijie; Du, Yijun; Feng, Wen-Hai

    2016-09-01

    Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) is one of the most important infectious diseases in swine industry. IL-15 is a pleiotropic cytokine and has been shown to be essential to transform NKs, CD8 T cells, and other cells of the immune systems into functional effectors. Here, we demonstrated that the broad-spectrum or conventional PKC inhibitors repressed PRRSV-induced IL-15 expression and NF-κB activation. Subsequently, we found that the PKCβ specific inhibitor inhibited PRRSV-induced IL-15 production, which was also confirmed by knock-down of PKCβ1, suggesting that PKCβ1 is involved in the PRRSV-induced IL-15 expression. In addition, we demonstrated that PRRSV activated NF-κB through PKCβ1-induced TAK1 activation. Finally, we demonstrated that PRRSV activated PKCβ1 dependent on the participation of TRIF and MAVS. These data indicate that PRRSV up-regulates IL-15 through TRIF/MAVS-PKCβ1-TAK1-NF-κB signaling pathway. These findings will provide new insights into the molecular mechanisms of IL-15 production induced by PRRSV. Copyright © 2016. Published by Elsevier Inc.

  1. Enhancing heterologous protection in pigs vaccinated with chimeric porcine reproductive and respiratory syndrome virus containing the full-length sequences of shuffled structural genes of multiple heterologous strains.

    Science.gov (United States)

    Tian, Debin; Cao, Dianjun; Lynn Heffron, C; Yugo, Danielle M; Rogers, Adam J; Overend, Christopher; Matzinger, Shannon R; Subramaniam, Sakthivel; Opriessnig, Tanja; LeRoith, Tanya; Meng, Xiang-Jin

    2017-04-25

    Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of arguably the most economically important global swine disease. The extensive genetic variation of PRRSV strains is a major obstacle for heterologous protection of current vaccines. Previously, we constructed a panel of chimeric viruses containing only the ectodomain sequences of DNA-shuffled structural genes of different PRRSV strains in the backbone of a commercial vaccine, and found that one chimeric virus had an improved cross-protection efficacy. In this present study, to further enhance the cross-protective efficacy against heterologous strains, we constructed a novel chimeric virus VR2385-S3456 containing the full-length sequences of shuffled structural genes (ORFs 3-6) from 6 heterologous PRRSV strains in the backbone of PRRSV strain VR2385. We showed that the chimeric virus VR2385-S3456 induced a high level of neutralizing antibodies in pigs against two heterologous strains. A subsequent vaccination and challenge study in 48 pigs revealed that the chimeric virus VR2385-S3456 conferred an enhanced cross-protection when challenged with heterologous virus strain NADC20 or a contemporary heterologous strain RFLP 1-7-4. The results suggest that the chimera VR2385-S3456 may be a good PRRSV vaccine candidate for further development to confer heterologous protection. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Systemic and mucosal immunity induced by attenuated Salmonella enterica serovar Typhimurium expressing ORF7 of porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Han, Young Woo; Kim, Seong Bum; Rahman, Masudur; Uyangaa, Erdenebileg; Lee, Byung Min; Kim, Jin Hyoung; Park, Ki In; Hong, Jin Tae; Han, Sang-Bae; Eo, Seong Kug

    2011-07-01

    Oral administration of attenuated Salmonella vaccine may provide valuable advantages such as low cost, easy preparation, and safety. Attenuated Salmonella vaccines also serve as carriers of foreign antigens and immunomodulatory cytokines. Presently, an attenuated Salmonella enterica serovar Typhimurium strain was used as a carrier for open reading frame 7 (ORF7) protein of porcine reproductive and respiratory syndrome virus (PRRSV), a swine pathogen of significant global economic importance. Initially, an attenuated S. enterica serovar Typhimurium expressing ORF7 gene derived from PRRSV Korean isolate was constructed. Following oral administration of a single dose of the attenuated Salmonella vaccine expressing PRRSV ORF7, humoral and cell-mediated immune responses specific for ORF7 were induced at both systemic and mucosal sites including spleen, mesenteric lymph node, Peyer's patch, and laminar propria, as evaluated by determining serum ORF7-specific IgG and mucosal IgA responses, as well as Th1- and Th2-type cytokine production from antigen-stimulated T cells. The induced humoral responses were sustained for at least 12weeks post-immunization. In particular, the immunized mice displayed immune responses to both the foreign ORF7 antigen and Salmonella itself. The results indicate the value of attenuated S. enterica serovar Typhimurium as an oral carrier of PRRSV antigenic proteins to induce effective systemic and mucosal immunity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. Comparison of protocols for the analysis of type 1 porcine reproductive and respiratory syndrome virus by RT-PCR using oral fluids.

    Science.gov (United States)

    Gibert, Elisa; Martín-Valls, Gerard; Mateu, Enric

    2017-05-01

    The detection of porcine reproductive and respiratory syndrome virus (PRRSV) in oral fluids (OF) by quantitative real-time polymerase chain reaction (qRT-PCR) is gaining increasing popularity. However, the different steps leading to a result have not been extensively evaluated. The aim of the present study was to examine the effect on the performance of qRT-PCR with different sampling materials, conditions of storage of the OF, the need for centrifuging OF, as well as to compare RNA extraction methods and PCR mixes. For the assays, pen-based oral fluids were used, which were pooled and spiked in a serial dilution (up to genotype 100 TCID50/mL) of type 1 PRRSV isolate 3267. Centrifugation at 15,000g for 15min resulted in an increase in sensitivity (1-2 PCR cycles) that was significant (PPCR Kit PCR mix reagents were more sensitive for the detection of PRRSV using a purified plasmid as standard, but LSI VetMAX PRRSV EU/NA PRRSV reagents resulted in a slightly better sensitivity with OF (pPCR. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Computer-aided codon-pairs deoptimization of the major envelope GP5 gene attenuates porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Ni, Yan-Yan; Zhao, Zhao; Opriessnig, Tanja; Subramaniam, Sakthivel; Zhou, Lei; Cao, Dianjun; Cao, Qian; Yang, Hanchun; Meng, Xiang-Jin

    2014-02-01

    Synthetic attenuated virus engineering (SAVE) is an emerging technology that enables rapid attenuation of viruses. In this study, by using SAVE we demonstrated rapid attenuation of an arterivirus, porcine reproductive and respiratory syndrome virus (PRRSV). The major envelope GP5 gene of PRRSV was codon-pair deoptimized aided by a computer algorithm. The codon-pair deoptimized virus, designated as SAVE5 with a deoptimized GP5 gene, was successfully rescued in vitro. The SAVE5 virus replicated at a lower level in vitro with a significant decrease of GP5 protein expression compared to the wild-type PRRSV VR2385 virus. Pigs experimentally infected with the SAVE5 virus had significantly lower viremia level up to 14 days post-infection as well as significantly reduced gross and histological lung lesions when compared to wild-type PRRSV VR2385 virus-infected pigs, indicating the attenuation of the SAVE5 virus. This study proved the feasibility of rapidly attenuating PRRSV by SAVE. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Pathologic Evaluation of Type 2 Porcine Reproductive and Respiratory Syndrome Virus Infection at the Maternal-Fetal Interface of Late Gestation Pregnant Gilts

    Science.gov (United States)

    Novakovic, Predrag; Harding, John C. S.; Al-Dissi, Ahmad N.; Ladinig, Andrea; Detmer, Susan E.

    2016-01-01

    The pathogenesis of fetal death caused by porcine reproductive and respiratory syndrome virus (PRRSV) remains unclear. The objective of this study was to improve our understanding of the pathogenesis by assessing potential relationships between specific histopathological lesions and PRRSV RNA concentration in the fetuses and the maternal-fetal interface. Pregnant gilts were inoculated with PRRSV (n = 114) or sham inoculated (n = 19) at 85±1 days of gestation. Dams and their litters were humanely euthanized and necropsied 21 days later. PRRSV RNA concentration was measured by qRT-PCR in the maternal-fetal interface and fetal thymus (n = 1391). Presence of fetal lesions was positively related to PRRSV RNA concentration in the maternal-fetal interface and fetal thymus (Pgilts induces significant histopathological lesions at maternal-fetal interface, but they are not associated with presence of PRRSV in the maternal-fetal interface at 21 days post infection. Conversely, fetal pathological lesions are associated with presence of PRRSV in the maternal-fetal interface and fetal thymus, and meconium staining is significantly associated with the presence of both fetal and umbilical lesions observed 21 days post infection. PMID:26963101

  6. Changes in leukocyte subsets of pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome.

    Science.gov (United States)

    Ladinig, Andrea; Gerner, Wilhelm; Saalmüller, Armin; Lunney, Joan K; Ashley, Carolyn; Harding, John C S

    2014-12-14

    In spite of more than two decades of extensive research, the understanding of porcine reproductive and respiratory syndrome virus (PRRSv) immunity is still incomplete. A PRRSv infection of the late term pregnant female can result in abortions, early farrowings, fetal death, and the birth of weak, congenitally infected piglets. The objectives of the present study were to investigate changes in peripheral blood mononuclear cell populations in third trimester pregnant females infected with type 2 PRRSv (NVSL 97-7895) and to analyze potential relationships with viral load and fetal mortality rate. PRRSv infection caused a massive, acute drop in total leukocyte counts affecting all PBMC populations by two days post infection. Except for B cells, cell counts started to rebound by day six post infection. Our data also show a greater decrease of naïve B cells, T-helper cells and cytolytic T cells than their respective effector or memory counterparts. Absolute numbers of T cells and γδ T cells were negatively associated with PRRSv RNA concentration in gilt serum over time. Additionally, absolute numbers of T helper cells may be predictive of fetal mortality rate. The preceding three leukocyte populations may therefore be predictive of PRRSv-related pathological outcomes in pregnant gilts. Although many questions regarding the immune responses remain unanswered, these findings provide insight and clues that may help reduce the impact of PRRSv in pregnant gilts.

  7. Inactivated porcine reproductive and respiratory syndrome virus vaccine adjuvanted with Montanide™ Gel 01 ST elicits virus-specific cross-protective inter-genotypic response in piglets.

    Science.gov (United States)

    Tabynov, Kairat; Sansyzbay, Abylay; Tulemissova, Zhanara; Tabynov, Kaissar; Dhakal, Santosh; Samoltyrova, Aigul; Renukaradhya, Gourapura J; Mambetaliyev, Muratbay

    2016-08-30

    The efficacy of a novel BEI-inactivated porcine reproductive and respiratory syndrome virus (PRRSV) candidate vaccine in pigs, developed at RIBSP Republic of Kazakhstan and delivered with an adjuvant Montanide™ Gel 01 ST (D/KV/ADJ) was compared with a commercial killed PRRSV vaccine (NVDC-JXA1, C/KV/ADJ) used widely in swine herds of the Republic of Kazakhstan. Clinical parameters (body temperature and respiratory disease scores), virological and immunological profiles [ELISA and virus neutralizing (VN) antibody titers], macroscopic lung lesions and viral load in the lungs (quantitative real-time PCR and cell culture assay) were assessed in vaccinated and both genotype 1 and 2 PRRSV challenged pigs. Our results showed that the commercial vaccine failed to protect pigs adequately against the clinical disease, viremia and lung lesions caused by the challenged field isolates, Kazakh strains of PRRSV type 1 and type 2 genotypes. In contrast, clinical protection, absence of viremia and lung lesions in D/KV/ADJ vaccinated pigs was associated with generation of VN antibodies in both homologous vaccine strain LKZ/2010 (PRRSV type 2) and a heterogeneous type 1 PRRSV strain (CM/08) challenged pigs. Thus, our data indicated the induction of cross-protective VN antibodies by D/KV/ADJ vaccine, and importantly demonstrated that an inactivated PRRSV vaccine could also induce cross-protective response across the viral genotype. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Comparison of self-processing of foot-and-mouth disease virus leader proteinase and porcine reproductive and respiratory syndrome virus leader proteinase nsp1α.

    Science.gov (United States)

    Steinberger, Jutta; Kontaxis, Georg; Rancan, Chiara; Skern, Tim

    2013-09-01

    The foot-and-mouth disease virus leader proteinase (Lb(pro)) cleaves itself off the nascent viral polyprotein. NMR studies on the monomeric variant Lb(pro) L200F provide structural evidence for intramolecular self-processing. (15)N-HSQC measurements of Lb(pro) L200F showed specifically shifted backbone signals in the active and substrate binding sites compared to the monomeric variant sLb(pro), lacking six C-terminal residues. This indicates transient intramolecular interactions between the C-terminal extension (CTE) of one molecule and its own active site. Contrastingly, the porcine reproductive and respiratory syndrome virus (PRRSV) leader proteinase nsp1α, with a papain-like fold like Lb(pro), stably binds its own CTE. Parts of the β-sheet domains but none of the α-helical domains of Lb(pro) and nsp1α superimpose; consequently, the α-helical domain of nsp1α is oriented differently relative to its β-sheet domain. This provides a large interaction surface for the CTE with the globular domain, stabilising the intramolecular complex. Consequently, self-processing inactivates nsp1α but not Lb(pro). Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Comparative analysis of apoptotic changes in peripheral immune organs and lungs following experimental infection of piglets with highly pathogenic and classical porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Wang, Gang; He, Yuli; Tu, Yabin; Liu, Yonggang; Zhou, En-Min; Han, Zifeng; Jiang, Chenggang; Wang, Shujie; Shi, Wenda; Cai, Xuehui

    2014-01-06

    Our previous studies have demonstrated that piglets infected with highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) may develop significant thymus atrophy, which related to thymocytes apoptosis. However, apart from that detected in the thymus, there are no reports describing cell apoptosis induced by HP-PRRSV infection. In this study, we analyzed comparatively the pathological changes, cell apoptosis and viral load in peripheral immune organs including tonsil, inguinal lymph nodes (ILNs) and spleen and lungs following experimental infection of piglets with HP-PRRSV HuN4 and classical PRRSV CH-1a. HP-PRRSV HuN4 exhibited much stronger cell tropism than CH-1a in immune organs and lungs of piglets. HuN4 infection led to the serious injuries in tonsils, ILNs, spleens and lungs, especially apoptosis in these organs was significant. HuN4 infection induced severe lesions (gross pathology, histopathology and cell apoptosis) in the peripheral immune organs and lungs of infected piglets. Large numbers of apoptotic cells in immune organs and lung induced by HuN4 may play a role in the pathogenesis of the HP-PRRS and the distinct injuries caused by HuN4 infection may be associated with the high mortality rate of HP-PRRS in pigs.

  10. Identification of epitopes on nonstructural protein 7 of porcine reproductive and respiratory syndrome virus recognized by monoclonal antibodies using phage-display technology.

    Science.gov (United States)

    Wang, Heng; Liu, Rongchang; Zhang, Weidong; Sun, Lingshuang; Ning, Zhangyong; Ji, Fangxiao; Cui, Jin; Zhang, Guihong

    2017-08-01

    Nonstructural protein 7 (nsp7) of porcine reproductive and respiratory syndrome virus (PRRSV) is considered to be a suitable reagent for the development of serological diagnostic assays. It can be expressed as a soluble recombinant protein in Escherichia coli, and its antibody response may continue up to 202 days post-infection. Furthermore, the region encoded by nsp7 is highly homologous among various strains within the genotype, and the results of nsp7-based enzyme-linked immunosorbent assay (ELISA) showed high agreement with previous Idexx ELISA results. All these evidences suggest the existence of important epitopes on nsp7, though the characteristics of these epitopes remain unclear. In the present study, we prepared three monoclonal antibodies against nsp7 protein and used them to screen the epitope-distribution characteristics of PRRSV nsp7 protein by phage-display technology. We identified a linear epitope NAWGDEDRLN at amino acids 153-162 type II PRRSV nsp7β subunit. This newly defined epitope showed excellent reactivity with PRSSV-positive serum samples. These results further our understanding of the antigenic structure of nsp7 protein, and provide efficient reagents for PRRSV serological tests.

  11. Porcine Reproductive and Respiratory Syndrome Virus Infection Induces Stress Granule Formation Depending on Protein Kinase R-like Endoplasmic Reticulum Kinase (PERK) in MARC-145 Cells.

    Science.gov (United States)

    Zhou, Yanrong; Fang, Liurong; Wang, Dang; Cai, Kaimei; Chen, Huanchun; Xiao, Shaobo

    2017-01-01

    Stress granules (SGs) are sites of mRNA storage that are formed in response to various conditions of stress, including viral infections. Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus that has been devastating the swine industry worldwide since the late 1980s. In this study, we found that infection of PRRSV strain WUH3 (genotype 2 PRRSV) induced stable formation of robust SGs in MARC-145 cells, as demonstrated by the recruitment of marker proteins of SGs, including TIA1, G3BP1, and eIF3η. Treatment with specific inhibitors or siRNAs against the stress kinases that are involved in SG formation revealed that PRRSV induced SG formation through a PERK (protein kinase R-like endoplasmic reticulum kinase)-dependent mechanism. Impairment of SG assembly by concomitant knockdown of the SG marker proteins (TIA1, G3BP1, and TIAR) did not affect PRRSV growth, while significantly enhanced PRRSV-induced NF-κB subunit p65 phosphorylation and inflammatory cytokine production. Taken together, our results demonstrate that PRRSV induces SG formation via a PERK-dependent pathway and that SGs are involved in the signaling pathway of the PRRSV-induced inflammatory response in MARC-145 cells.

  12. Activation of c-Jun NH(2)-terminal kinase is required for porcine reproductive and respiratory syndrome virus-induced apoptosis but not for virus replication.

    Science.gov (United States)

    Yin, Shutao; Huo, Yazhen; Dong, Yinhui; Fan, Lihong; Yang, Hanchun; Wang, Leyuan; Ning, Yibao; Hu, Hongbo

    2012-06-01

    Apoptosis of host cells plays a critical role in pathogenesis of virus infection. MAPK kinases especially stress-activated protein kinases c-Jun NH(2)-terminal kinase (SAPK/JNK) and p38 are often involved in virus-mediated apoptosis. It has been shown that porcine reproductive and respiratory syndrome virus (PRRSV) infection resulted in apoptosis of the host cells both in vitro and in vivo. The current investigation was initiated to determine whether stress-activated protein kinases JNK and p38 play a role in apoptosis induction by PRRSV infection. We examined phosphorylation of JNK and p38, and found that JNK but not p38 was activated in response to PRRSV infection. We then examined effects of this kinase on apoptosis induction and virus replication by using specific inhibitor. We found that JNK inhibition by its inhibitor SP600125 led to the abolishment of PRRSV-mediated apoptosis, but did not suppress virus replication. Further studies demonstrated that ROS generation was involved in JNK activation, and Bcl-2 family anti-apoptotic proteins Mcl-1 and Bcl-xl were downstream targets of JNK to mediate apoptosis. We conclude that activation of JNK signaling pathway is essential for PRRSV-mediated apoptosis but not for virus replication. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus

    Science.gov (United States)

    2010-01-01

    Background Prompt detection of PRRSV in the field samples is important for effective PRRS control, thereby reducing the potentially serious economic damage which can result from an outbreak. In this study, a rapid SYBR-based, one step real-time RT-PCR quantitative reverse transcription PCR (qRT-PCR) has been developed for the detection of porcine reproductive and respiratory syndrome virus (PRRSV). Primers were designed based on the sequence of highly conservative region of PRRSV N gene. Results The sensitivity of the real-time qRT-PCR assay was achieved through PRRSV ch-1a RNA for the generation of a standard curve. The detection limit of the assay was found to be 9.6 RNA copies per reaction mixture. This assay had excellent intra- and inter-assay reproducibility as in total 65 field samples were screened for the presence of PRRSV by conventional RT-PCR in parallel with qRT-PCR, and the detection rate increased from 60.0% to 76.9%. Moreover, the specificity result indicated that this assay could reliably differentiate PRRSV from the other swine viral diseases, such as classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV). Conclusion The real-time qRT-PCR assay described in this report allows the rapid, specific and sensitive laboratory detection of PRRSV in field samples. PMID:20459705

  14. The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus

    Directory of Open Access Journals (Sweden)

    Liu XiangTao

    2010-05-01

    Full Text Available Abstract Background Prompt detection of PRRSV in the field samples is important for effective PRRS control, thereby reducing the potentially serious economic damage which can result from an outbreak. In this study, a rapid SYBR-based, one step real-time RT-PCR quantitative reverse transcription PCR (qRT-PCR has been developed for the detection of porcine reproductive and respiratory syndrome virus (PRRSV. Primers were designed based on the sequence of highly conservative region of PRRSV N gene. Results The sensitivity of the real-time qRT-PCR assay was achieved through PRRSV ch-1a RNA for the generation of a standard curve. The detection limit of the assay was found to be 9.6 RNA copies per reaction mixture. This assay had excellent intra- and inter-assay reproducibility as in total 65 field samples were screened for the presence of PRRSV by conventional RT-PCR in parallel with qRT-PCR, and the detection rate increased from 60.0% to 76.9%. Moreover, the specificity result indicated that this assay could reliably differentiate PRRSV from the other swine viral diseases, such as classical swine fever virus (CSFV, swine vesicular disease virus (SVDV and vesicular exanthema of swine virus (VESV. Conclusion The real-time qRT-PCR assay described in this report allows the rapid, specific and sensitive laboratory detection of PRRSV in field samples.

  15. The effects of the context-dependent codon usage bias on the structure of the nsp1α of porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Ding, Yao-zhong; You, Ya-nan; Sun, Dong-jie; Chen, Hao-tai; Wang, Yong-lu; Chang, Hui-yun; Pan, Li; Fang, Yu-zhen; Zhang, Zhong-wang; Zhou, Peng; Lv, Jian-liang; Liu, Xin-sheng; Shao, Jun-jun; Zhao, Fu-rong; Lin, Tong; Stipkovits, Laszlo; Pejsak, Zygmunt; Zhang, Yong-guang; Zhang, Jie

    2014-01-01

    The information about the crystal structure of porcine reproductive and respiratory syndrome virus (PRRSV) leader protease nsp1α is available to analyze the roles of tRNA abundance of pigs and codon usage of the nsp1 α gene in the formation of this protease. The effects of tRNA abundance of the pigs and the synonymous codon usage and the context-dependent codon bias (CDCB) of the nsp1 α on shaping the specific folding units (α-helix, β-strand, and the coil) in the nsp1α were analyzed based on the structural information about this protease from protein data bank (PDB: 3IFU) and the nsp1 α of the 191 PRRSV strains. By mapping the overall tRNA abundance along the nsp1 α, we found that there is no link between the fluctuation of the overall tRNA abundance and the specific folding units in the nsp1α, and the low translation speed of ribosome caused by the tRNA abundance exists in the nsp1 α. The strong correlation between some synonymous codon usage and the specific folding units in the nsp1α was found, and the phenomenon of CDCB exists in the specific folding units of the nsp1α. These findings provide an insight into the roles of the synonymous codon usage and CDCB in the formation of PRRSV nsp1α structure.

  16. The Effects of the Context-Dependent Codon Usage Bias on the Structure of the nsp1α of Porcine Reproductive and Respiratory Syndrome Virus

    Directory of Open Access Journals (Sweden)

    Yao-zhong Ding

    2014-01-01

    Full Text Available The information about the crystal structure of porcine reproductive and respiratory syndrome virus (PRRSV leader protease nsp1α is available to analyze the roles of tRNA abundance of pigs and codon usage of the nsp1α gene in the formation of this protease. The effects of tRNA abundance of the pigs and the synonymous codon usage and the context-dependent codon bias (CDCB of the nsp1α on shaping the specific folding units (α-helix, β-strand, and the coil in the nsp1α were analyzed based on the structural information about this protease from protein data bank (PDB: 3IFU and the nsp1α of the 191 PRRSV strains. By mapping the overall tRNA abundance along the nsp1α, we found that there is no link between the fluctuation of the overall tRNA abundance and the specific folding units in the nsp1α, and the low translation speed of ribosome caused by the tRNA abundance exists in the nsp1α. The strong correlation between some synonymous codon usage and the specific folding units in the nsp1α was found, and the phenomenon of CDCB exists in the specific folding units of the nsp1α. These findings provide an insight into the roles of the synonymous codon usage and CDCB in the formation of PRRSV nsp1α structure.

  17. Predicted Peptides from Non-Structural Proteins of Porcine Reproductive and Respiratory Syndrome Virus Are Able to Induce IFN-γ and IL-10

    Directory of Open Access Journals (Sweden)

    Enric Mateu

    2013-02-01

    Full Text Available This work describes peptides from non-structural proteins (nsp of porcine reproductive and respiratory syndrome virus (PRRSV predicted as potential T cell epitopes by bioinfornatics and tested for their ability to induce IFN-γ and IL-10 responses. Pigs immunized with either genotype 1 or genotype 2 PRRSV attenuated vaccines (n=5/group and unvaccinated pigs (n = 4 were used to test the peptides. Swine leukocyte antigen haplotype of each pig was also determined. Pigs were initially screened for IFN-γ responses (ELISPOT and three peptides were identified; two of them in non-conserved segments of nsp2 and nsp5 and the other in a conserved region of nsp5 peptide. Then, peptides were screened for IL-10 inducing properties. Six peptides were found to induce IL-10 release in PBMC and some of them were also able to inhibit IFN-γ responses on PHA-stimulated cells. Interestingly, the IFN-γ low responder pigs against PRRSV were mostly homozygous for their SLA haplotypes. In conclusion, these results indicate that nsp of PRRSV contain T-cell epitopes inducing IFN-γ responses as well as IL-10 inducing segments with inhibitory capabilities.

  18. Unraveling the contact patterns and network structure of pig shipments in the United States and its association with porcine reproductive and respiratory syndrome virus (PRRSV) outbreaks.

    Science.gov (United States)

    Lee, Kyuyoung; Polson, Dale; Lowe, Erin; Main, Rodger; Holtkamp, Derald; Martínez-López, Beatriz

    2017-03-01

    The analysis of the pork value chain is becoming key to understanding the risk of infectious disease dissemination in the swine industry. In this study, we used social network analysis to characterize the swine shipment network structure and properties in a typical multisite swine production system in the US. We also aimed to evaluate the association between network properties and porcine respiratory and reproductive syndrome virus (PRRSV) transmission between production sites. We analyzed the 109,868 swine shipments transporting over 93 million swine between more than 500 production sites from 2012 to 2014. A total of 248 PRRSV positive occurrences were reported from 79 production sites during those 3 years. The temporal dynamics of swine shipments was evaluated by computing network properties in one-month and three-month networks. The association of PRRS occurrence in sow farms with centrality properties from one-month and three-month networks was assessed by using the multilevel logistic regression. All monthly networks showed a scale-free network topology with positive degree assortativity. The regression model revealed that out-degree centrality had a negative association with PRRS occurrence in sow farms in both one-month and three-month networks [OR=0.79 (95% CI, 0.63-0.99) in one-month network and 0.56 (95% CI, 0.36, 0.88) in three-month network] and in-closeness centrality model was positively associated with PRRS occurrence in sow farms in the three-month network [OR=2.45 (95% CI, 1.14-5.26)]. We also describe how the occurrence of porcine epidemic diarrheac (PED) outbreaks severely affected the network structure as well as the PRRS occurrence reports and its association with centrality measures in sow farms. The structure of the swine shipment network and the connectivity between production sites influenced on the PRRSV transmission. The use of network topology and characteristics combining with spatial analysis based on fine scale geographical location

  19. Carbon Monoxide Inhibits Porcine Reproductive and Respiratory Syndrome Virus Replication by the Cyclic GMP/Protein Kinase G and NF-κB Signaling Pathway.

    Science.gov (United States)

    Zhang, Angke; Zhao, Lijuan; Li, Na; Duan, Hong; Liu, Hongliang; Pu, Fengxing; Zhang, Gaiping; Zhou, En-Min; Xiao, Shuqi

    2017-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant economic losses to the pork industry worldwide each year. Our previous research demonstrated that heme oxygenase-1 (HO-1) can suppress PRRSV replication via an unknown molecular mechanism. In this study, inhibition of PRRSV replication was demonstrated to be mediated by carbon monoxide (CO), a downstream metabolite of HO-1. Using several approaches, we demonstrate that CO significantly inhibited PRRSV replication in both a PRRSV permissive cell line, MARC-145, and the predominant cell type targeted during in vivo PRRSV infection, porcine alveolar macrophages (PAMs). Our results showed that CO inhibited intercellular spread of PRRSV; however, it did not affect PRRSV entry into host cells. Furthermore, CO was found to suppress PRRSV replication via the activation of the cyclic GMP/protein kinase G (cGMP/PKG) signaling pathway. CO significantly inhibits PRRSV-induced NF-κB activation, a required step for PRRSV replication. Moreover, CO significantly reduced PRRSV-induced proinflammatory cytokine mRNA levels. In conclusion, the present study demonstrates that CO exerts its anti-PRRSV effect by activating the cellular cGMP/PKG signaling pathway and by negatively regulating cellular NF-κB signaling. These findings not only provide new insights into the molecular mechanism of HO-1 inhibition of PRRSV replication but also suggest potential new control measures for future PRRSV outbreaks. PRRSV causes great economic losses each year to the swine industry worldwide. Carbon monoxide (CO), a metabolite of HO-1, has been shown to have antimicrobial and antiviral activities in infected cells. Our previous research demonstrated that HO-1 can suppress PRRSV replication. Here we show that endogenous CO produced through HO-1 catalysis mediates the antiviral effect of HO-1. CO inhibits PRRSV replication by activating the cellular cGMP/PKG signaling pathway and by negatively regulating cellular NF

  20. Reproductive, productivity, and mortality outcomes in late-gestation gilts and their litters following simulation of inadvertent exposure to a modified-live vaccine strain of porcine reproductive and respiratory syndrome (PRRS) virus.

    Science.gov (United States)

    Schelkopf, Adam; Nerem, Joel; Cowles, Bobby; Amodie, Deb; Swalla, Richard; Dee, Scott

    2014-08-06

    The study evaluated the safety of a modified live-virus (MLV) porcine reproductive and respiratory syndrome (PRRS) vaccine in susceptible, pregnant gilts. To simulate inadvertent exposure secondary to postvaccination shedding of PRRS-MLV, seronegative gilts (n=51) were exposed by IM vaccination at 90 days of gestation. Vaccinated and nonvaccinated, seronegative control gilts (n=25) were maintained in separate facilities. The PRRS-MLV vaccine was given in a 2mL dose on day 0. On day 7 all vaccinated gilts were PRRSV-PCR-positive for PRRSV and had responded serologically as determined by an ELISA. All control gilts remained PRRSV-PCR- and ELISA-negative throughout the study. Abortions did not occur in gilts from either group. The difference between vaccinated and control gilts in average number of piglets per litter (12.43 and 12.16, respectively), number of live births per litter (11.21 and 11.54), and mean piglet birth weight (3.22 and 3.26 lbs) were not significantly different. Piglets in the control group had significantly greater average daily gain versus piglets from vaccinated gilts (0.52 vs. 0.46 lbs, Pgilts (19.7% vs. 10.9%). A single gilt accounted for 18.2% of stillbirths in the vaccinated group. Air samples were borderline PRRSV-PCR-positive for PRRSV on days 29 and 32, after more than 98% of gilts had farrowed. Results demonstrated that vaccination of pregnant gilts at the time of peak fetal susceptibility was non-abortigenic and that the PRRS-MLV agent did not significantly affect reproductive outcomes. Lower ADG in piglets from vaccinated gilts may be due to PRRS-MLV viremia following transplacental or post-farrowing exposure. Air sampling results indicated that environmental contamination with PRRS-MLV shed from vaccinated gilts was minimal. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Type 2 porcine reproductive and respiratory syndrome virus infection increases apoptosis at the maternal-fetal interface in late gestation pregnant gilts.

    Directory of Open Access Journals (Sweden)

    Predrag Novakovic

    Full Text Available The pathogenesis of fetal death associated with porcine reproductive and respiratory syndrome (PRRS is hypothesized to be a consequence of PRRS virus-induced apoptosis at the maternal-fetal interface (MFI. The objectives of this study were to evaluate distribution and degree of apoptosis in the uterine and fetal placental tissues during the experimental type 2 PRRS virus (PRRSV infection and determine associations between apoptosis at the MFI, PRRSV RNA concentration and antigen staining intensity, PRRSV-induced microscopic lesions, and fetal preservation status. A total of 114 naïve, high-health pregnant gilts were inoculated with type 2 PRRSV on gestation day 85±1 with euthanasia 21 days later; 19 sham-inoculated gilts served as controls. Two hundred and fifty samples of uterine tissue with fetal placenta were selected based on negative, low PRRSV RNA, and high PRRSV RNA concentration (0, 2.7 log10 copies/mg, respectively. TUNEL assay was used to detect apoptosis in the endometrium and at the MFI. PRRSV RNA concentration and numbers of PRRSV immunopositive cells in uterine and placental tissue were positively associated with the severity of apoptosis in the endometrium and the MFI (P<0.001, P<0.05 and P<0.001, respectively. The number of TUNEL positive cells at the MFI was also positively associated with the severity (P<0.001 of vasculitis, but not total numbers of inflammatory cells in the endometrium. Increased numbers of TUNEL positive cells at the MFI were associated with PRRSV load in the fetal thymus, and greater odds of meconium staining of the fetus at 21 days post infection (P<0.001 for both. These findings suggest an important role of apoptosis in the pathogenesis of uterine epithelial and trophoblastic cell death at the MFI. Moreover, apoptosis at the MFI is significantly associated with fetal demise during in utero type 2 PRRSV infection.

  2. Comparison of protection provided by type 1 and type 2 porcine reproductive and respiratory syndrome field viruses against homologous and heterologous challenge.

    Science.gov (United States)

    Choi, Kyuhyung; Park, Changhoon; Jeong, Jiwoon; Chae, Chanhee

    2016-08-15

    The objective of this study was to compare protection provided by type 1 and type 2 porcine reproductive and respiratory syndrome virus (PRRSV) against homologous and heterologous challenge based on clinical, virological, immunological, and pathological analysis. At 3 and 8 weeks of age, pigs were inoculated intranasally with either 3mL of tissue culture fluid containing 10(5) TCID50/mL of type 1 PRRSV or 3mL of tissue culture fluid containing 10(5) TCID50/mL of type 2 PRRSV. The homologous challenges resulted in a significant boost of the neutralizing antibodies (NA) and interferon-γ secreting cells (IFN-γ-SC) compared to heterologous challenges. The reduction of secondary challenging PRRSV viremia coincided with the appearance of homologous PRRSV-specific NA and IFN-γ-SC. Homologous challenge reduced the severity of lung lesions and levels of PRRSV viremia significantly in pigs in comparison with heterologous challenge. The differences in homologous and heterologous NA and IFN-γ-SC response may explain the differences in protection against homologous and heterologous challenge between type 1 and type 2 PRRSV. Primary challenge (immunization) with type 1 PRRSV provided protection against the secondary homologous challenge with type 1 PRRSV but failed to provide protection against the secondary heterologous challenge of type 2 PRRSV. Primary challenge with type 2 PRRSV provided protection against both the secondary homologous challenge with type 2 PRRSV and the secondary heterologous challenge with type 1 PRRSV. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Virus replicon particles expressing porcine reproductive and respiratory syndrome virus proteins elicit immune priming but do not confer protection from viremia in pigs.

    Science.gov (United States)

    Eck, Melanie; Durán, Margarita García; Ricklin, Meret E; Locher, Samira; Sarraseca, Javier; Rodríguez, María José; McCullough, Kenneth C; Summerfield, Artur; Zimmer, Gert; Ruggli, Nicolas

    2016-02-19

    Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of one of the most devastating and economically significant viral disease of pigs worldwide. The vaccines currently available on the market elicit only limited protection. Recombinant vesicular stomatitis virus (VSV) replicon particles (VRP) have been used successfully to induce protection against influenza A virus (IAV) in chickens and bluetongue virus in sheep. In this study, VSV VRP expressing the PRRSV envelope proteins GP5, M, GP4, GP3, GP2 and the nucleocapsid protein N, individually or in combination, were generated and evaluated as a potential vector vaccine against PRRSV infection. High level expression of the recombinant PRRSV proteins was demonstrated in cell culture. However, none of the PRRSV antigens expressed from VRP, with the exception of the N protein, did induce any detectable antibody response in pigs before challenge infection with PRRSV. After challenge however, the antibody responses against GP5, GP4 and GP3 appeared in average 2 weeks earlier than in pigs vaccinated with the empty control VRP. No reduction of viremia was observed in the vaccinated group compared with the control group. When pigs were co-vaccinated with VRP expressing IAV antigens and VRP expressing PRRSV glycoproteins, only antibody responses to the IAV antigens were detectable. These data show that the VSV replicon vector can induce immune responses to heterologous proteins in pigs, but that the PRRSV envelope proteins expressed from VSV VRP are poorly immunogenic. Nevertheless, they prime the immune system for significantly earlier B-cell responses following PRRSV challenge infection.

  4. Attempts to enhance cross-protection against porcine reproductive and respiratory syndrome viruses using chimeric viruses containing structural genes from two antigenically distinct strains.

    Science.gov (United States)

    Sun, Dong; Khatun, Amina; Kim, Won-Il; Cooper, Vickie; Cho, Yong-Il; Wang, Chong; Choi, Eun-Jin; Yoon, Kyoung-Jin

    2016-08-05

    Due to significant antigenic variations between field isolates of porcine reproductive and respiratory syndrome virus (PRRSV), suboptimal cross-protection between different viruses impedes the effective control of PRRS via vaccination. Our previous study showed that chimeric viruses containing mixed structural genes from two distinct strains (VR2332 and JA142) of PRRSV were highly susceptible to the viral neutralizing activity of antisera generated against both parental strains. In this study, three chimeric viruses (JAP5, JAP56 and JAP2-6) were constructed by replacing ORF5, ORFs 5 and 6, and ORFs 2-6 of VR2332 with the corresponding genes of JA142, respectively, and their ability to confer cross-protection against challenge with the VR2332 and JA142 strains was evaluated in vivo. A total of 114 pigs were divided into 6 groups, and each group was intramuscularly injected with one of the 3 chimeric viruses (n=16 pigs per group), VR2332 (n=24), JA142 (n=24), or sham inoculum (n=18). At 44days post-inoculation (dpi), these pigs were further divided into 15 groups (n=6 or 8 pigs per group) and intranasally challenged with VR2332, JA142, or sham inoculum. All pigs inoculated with one of the chimeric viruses prior to challenge had lower viremia levels than the challenge control pigs. Prior inoculation with JAP56 markedly decreased viremia to nearly undetectable levels in pigs challenged with either VR2332 or JA142. These results suggest that chimeric viruses harboring mixed structural genes from two distinct PRRSV strains can provide protection against both donor viruses. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Evaluation of the Cross-Protective Efficacy of a Chimeric Porcine Reproductive and Respiratory Syndrome Virus Constructed Based on Two Field Strains.

    Science.gov (United States)

    Shabir, Nadeem; Khatun, Amina; Nazki, Salik; Kim, Bumseok; Choi, Eun-Jin; Sun, Dong; Yoon, Kyoung-Jin; Kim, Won-Il

    2016-08-22

    One of the major hurdles to porcine reproductive and respiratory syndrome (PRRS) vaccinology is the limited or no cross-protection conferred by current vaccines. To overcome this challenge, a PRRS chimeric virus (CV) was constructed using an FL12-based cDNA infectious clone in which open reading frames (ORFs) 3-4 and ORFs 5-6 were replaced with the two Korean field isolates K08-1054 and K07-2273,respectively. This virus was evaluated as a vaccine candidate to provide simultaneous protection against two genetically distinct PRRS virus (PRRSV) strains. Thirty PRRS-negative three-week-old pigs were divided into five groups and vaccinated with CV, K08-1054, K07-2273, VR-2332, or a mock inoculum. At 25 days post-vaccination (dpv), the pigs in each group were divided further into two groups and challenged with either K08-1054 or K07-2273. All of the pigs were observed until 42 dpv and were euthanized for pathological evaluation. Overall, the CV-vaccinated group exhibited higher levels of tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), and interleukin-12 (IL-12) expression and of serum virus-neutralizing antibodies compared with the other groups after vaccination and also demonstrated better protection levels against both viruses compared with the challenge control group. Based on these results, it was concluded that CV might be an effective vaccine model that can confer a broader range of cross-protection to various PRRSV strains.

  6. Comparison of self-processing of foot-and-mouth disease virus leader proteinase and porcine reproductive and respiratory syndrome virus leader proteinase nsp1α

    Energy Technology Data Exchange (ETDEWEB)

    Steinberger, Jutta [Max F. Perutz Laboratories, Medical University of Vienna, Department of Medical Biochemistry, Dr. Bohr-Gasse 9/3, A-1030 Vienna (Austria); Kontaxis, Georg [Max F. Perutz Laboratories, University of Vienna, Department of Structural and Computational Biology, Campus Vienna Biocenter 5, A-1030 Vienna (Austria); Rancan, Chiara [Helmholtz Zentrum München, Department of Gene Vectors, Haematologikum, Marchioninistrasse 25, D-81377 Munich (Germany); Skern, Tim, E-mail: timothy.skern@meduniwien.ac.at [Max F. Perutz Laboratories, Medical University of Vienna, Department of Medical Biochemistry, Dr. Bohr-Gasse 9/3, A-1030 Vienna (Austria)

    2013-09-01

    The foot-and-mouth disease virus leader proteinase (Lb{sup pro}) cleaves itself off the nascent viral polyprotein. NMR studies on the monomeric variant Lb{sup pro} L200F provide structural evidence for intramolecular self-processing. {sup 15}N-HSQC measurements of Lb{sup pro} L200F showed specifically shifted backbone signals in the active and substrate binding sites compared to the monomeric variant sLb{sup pro}, lacking six C-terminal residues. This indicates transient intramolecular interactions between the C-terminal extension (CTE) of one molecule and its own active site. Contrastingly, the porcine reproductive and respiratory syndrome virus (PRRSV) leader proteinase nsp1α, with a papain-like fold like Lb{sup pro}, stably binds its own CTE. Parts of the β-sheet domains but none of the α-helical domains of Lb{sup pro} and nsp1α superimpose; consequently, the α-helical domain of nsp1α is oriented differently relative to its β-sheet domain. This provides a large interaction surface for the CTE with the globular domain, stabilising the intramolecular complex. Consequently, self-processing inactivates nsp1α but not Lb{sup pro}. - Highlights: • We examine self-processing of the leader protease of foot-and-mouth disease virus. • NMR analysis strongly supports intramolecular self-processing. • Self-processing is a dynamic process with no stable complex. • Structural comparison with nsp1α of PRRSV which forms stable intramolecular complex. • Subdomain orientation explains differences in stability of intramolecular complexes.

  7. An evaluation of disinfectants for the sanitation of porcine reproductive and respiratory syndrome virus-contaminated transport vehicles at cold temperatures.

    Science.gov (United States)

    Dee, Scott; Deen, John; Burns, Danny; Douthit, George; Pijoan, Carlos

    2005-01-01

    The objective of this study was to evaluate the efficacy of commercially available disinfectants to sanitize porcine reproductive and respiratory syndrome virus (PRRSV) contaminated trailer models in cold climates (-20 degrees C and 4 degrees C). Disinfectants evaluated included Synergize, Aseptol 2000, Biophene, Sentramax, Virkon, Tek Trol, and DC&R. All products were applied to trailers via fumigation at 4 degrees C. Following experimental contamination of model trailers with PRRSV MN 30-100 (5 x 10(5) TCID50), models were tested for the presence or absence of PRRSV-RNA by polymerase chain reaction (PCR) on swabs collected 0, 30, and 60 min after treatment. Treatments included washing only, washing plus disinfectant fumigation, washing plus fumigation, and washing plus overnight drying. The PRRSV-RNA detected across trailers ranged from 0/12 replicates in trailers treated with Synergize or allowed to dry for 8 h. These trailers were also negative for the presence of infectious PRRSV, based on the lack of sentinel pig infection (0/4 replicates). In contrast, the detection of PRRSV-positive swabs by PCR ranged from 3/12 (Aseptol) to 10/12 (Biophene). Based on these results, the efficacy of Synergize was evaluated at -20 degrees C. In an attempt to reduce the impact of freezing on disinfectant activity, 30 mL of disinfectant was added to a 3840 mL of a 40% methanol solution, a 10% propylene glycol (PG) solution, or water alone. The PRRSV-contaminated trailers were treated with 1 of 3 disinfectant mixtures via fumigation, stored for 8 h at -20 degrees C, allowed to thaw, and sampled as described. Trailers treated with 40% methanol or 10% PG did not freeze and were negative for PRRSV-RNA and infectious virus following thawing. In contrast, trailers treated with disinfectant and water were frozen within 60 min at -20 degrees C, and decontamination was not successful.

  8. Estimating Parameters Related to the Lifespan of Passively Transferred and Vaccine-Induced Porcine Reproductive and Respiratory Syndrome Virus Type I Antibodies by Modeling Field Data

    Directory of Open Access Journals (Sweden)

    Mathieu Andraud

    2018-01-01

    Full Text Available The outputs of epidemiological models are strongly related to the structure of the model and input parameters. The latter are defined by fitting theoretical concepts to actual data derived from field or experimental studies. However, some parameters may remain difficult to estimate and are subject to uncertainty or sensitivity analyses to determine their variation range and their global impact on model outcomes. As such, the evaluation of immunity duration is often a puzzling issue requiring long-term follow-up data that are, most of time, not available. The present analysis aims at characterizing the kinetics of antibodies against Porcine Reproductive and Respiratory Syndrome virus (PRRSv from longitudinal data sets. The first data set consisted in the serological follow-up of 22 vaccinated gilts during 21 weeks post-vaccination (PV. The second one gathered the maternally derived antibodies (MDAs kinetics in piglets from three different farms up to 14 weeks of age. The peak of the PV serological response against PRRSv was reached 6.9 weeks PV on average with an average duration of antibodies persistence of 26.5 weeks. In the monitored cohort of piglets, the duration of passive immunity was found relatively short, with an average duration of 4.8 weeks. The level of PRRSv-MDAs was found correlated with the dams’ antibody titer at birth, and the antibody persistence was strongly related to the initial MDAs titers in piglets. These results evidenced the importance of PRRSv vaccination schedule in sows, to optimize the delivery of antibodies to suckling piglets. These estimates of the duration of active and passive immunity could be further used as input parameters of epidemiological models to analyze their impact on the persistence of PRRSv within farms.

  9. Type 2 porcine reproductive and respiratory syndrome virus infection increases apoptosis at the maternal-fetal interface in late gestation pregnant gilts

    Science.gov (United States)

    Harding, John C. S.; Al-Dissi, Ahmad N; Detmer, Susan E.

    2017-01-01

    The pathogenesis of fetal death associated with porcine reproductive and respiratory syndrome (PRRS) is hypothesized to be a consequence of PRRS virus-induced apoptosis at the maternal-fetal interface (MFI). The objectives of this study were to evaluate distribution and degree of apoptosis in the uterine and fetal placental tissues during the experimental type 2 PRRS virus (PRRSV) infection and determine associations between apoptosis at the MFI, PRRSV RNA concentration and antigen staining intensity, PRRSV-induced microscopic lesions, and fetal preservation status. A total of 114 naïve, high-health pregnant gilts were inoculated with type 2 PRRSV on gestation day 85±1 with euthanasia 21 days later; 19 sham-inoculated gilts served as controls. Two hundred and fifty samples of uterine tissue with fetal placenta were selected based on negative, low PRRSV RNA, and high PRRSV RNA concentration (0, 2.7 log10 copies/mg, respectively). TUNEL assay was used to detect apoptosis in the endometrium and at the MFI. PRRSV RNA concentration and numbers of PRRSV immunopositive cells in uterine and placental tissue were positively associated with the severity of apoptosis in the endometrium and the MFI (P<0.001, P<0.05 and P<0.001, respectively). The number of TUNEL positive cells at the MFI was also positively associated with the severity (P<0.001) of vasculitis, but not total numbers of inflammatory cells in the endometrium. Increased numbers of TUNEL positive cells at the MFI were associated with PRRSV load in the fetal thymus, and greater odds of meconium staining of the fetus at 21 days post infection (P<0.001 for both). These findings suggest an important role of apoptosis in the pathogenesis of uterine epithelial and trophoblastic cell death at the MFI. Moreover, apoptosis at the MFI is significantly associated with fetal demise during in utero type 2 PRRSV infection. PMID:28253336

  10. Estimating Parameters Related to the Lifespan of Passively Transferred and Vaccine-Induced Porcine Reproductive and Respiratory Syndrome Virus Type I Antibodies by Modeling Field Data.

    Science.gov (United States)

    Andraud, Mathieu; Fablet, Christelle; Renson, Patricia; Eono, Florent; Mahé, Sophie; Bourry, Olivier; Rose, Nicolas

    2018-01-01

    The outputs of epidemiological models are strongly related to the structure of the model and input parameters. The latter are defined by fitting theoretical concepts to actual data derived from field or experimental studies. However, some parameters may remain difficult to estimate and are subject to uncertainty or sensitivity analyses to determine their variation range and their global impact on model outcomes. As such, the evaluation of immunity duration is often a puzzling issue requiring long-term follow-up data that are, most of time, not available. The present analysis aims at characterizing the kinetics of antibodies against Porcine Reproductive and Respiratory Syndrome virus (PRRSv) from longitudinal data sets. The first data set consisted in the serological follow-up of 22 vaccinated gilts during 21 weeks post-vaccination (PV). The second one gathered the maternally derived antibodies (MDAs) kinetics in piglets from three different farms up to 14 weeks of age. The peak of the PV serological response against PRRSv was reached 6.9 weeks PV on average with an average duration of antibodies persistence of 26.5 weeks. In the monitored cohort of piglets, the duration of passive immunity was found relatively short, with an average duration of 4.8 weeks. The level of PRRSv-MDAs was found correlated with the dams' antibody titer at birth, and the antibody persistence was strongly related to the initial MDAs titers in piglets. These results evidenced the importance of PRRSv vaccination schedule in sows, to optimize the delivery of antibodies to suckling piglets. These estimates of the duration of active and passive immunity could be further used as input parameters of epidemiological models to analyze their impact on the persistence of PRRSv within farms.

  11. Evaluation of Control Strategies for Porcine Reproductive and Respiratory Syndrome (PRRS) in Swine Breeding Herds Using a Discrete Event Agent-Based Model.

    Science.gov (United States)

    Arruda, Andréia Gonçalves; Friendship, Robert; Carpenter, Jane; Greer, Amy; Poljak, Zvonimir

    2016-01-01

    The objective of this study was to develop a discrete event agent-based stochastic model to explore the likelihood of the occurrence of porcine reproductive and respiratory syndrome (PRRS) outbreaks in swine herds with different PRRS control measures in place. The control measures evaluated included vaccination with a modified-live attenuated vaccine and live-virus inoculation of gilts, and both were compared to a baseline scenario where no control measures were in place. A typical North American 1,000-sow farrow-to-wean swine herd was used as a model, with production and disease parameters estimated from the literature and expert opinion. The model constructed herein was not only able to capture individual animal heterogeneity in immunity to and shedding of the PRRS virus, but also the dynamic animal flow and contact structure typical in such herds under field conditions. The model outcomes included maximum number of females infected per simulation, and time at which that happened and the incidence of infected weaned piglets during the first year of challenge-virus introduction. Results showed that the baseline scenario produced a larger percentage of simulations resulting in outbreaks compared to the control scenarios, and interestingly some of the outbreaks occurred over long periods after virus introduction. The live-virus inoculation scenario showed promising results, with fewer simulations resulting in outbreaks than the other scenarios, but the negative impacts of maintaining a PRRS-positive population should be considered. Finally, under the assumptions of the current model, neither of the control strategies prevented the infection from spreading to the piglet population, which highlights the importance of maintaining internal biosecurity practices at the farrowing room level.

  12. Estimating Parameters Related to the Lifespan of Passively Transferred and Vaccine-Induced Porcine Reproductive and Respiratory Syndrome Virus Type I Antibodies by Modeling Field Data

    Science.gov (United States)

    Andraud, Mathieu; Fablet, Christelle; Renson, Patricia; Eono, Florent; Mahé, Sophie; Bourry, Olivier; Rose, Nicolas

    2018-01-01

    The outputs of epidemiological models are strongly related to the structure of the model and input parameters. The latter are defined by fitting theoretical concepts to actual data derived from field or experimental studies. However, some parameters may remain difficult to estimate and are subject to uncertainty or sensitivity analyses to determine their variation range and their global impact on model outcomes. As such, the evaluation of immunity duration is often a puzzling issue requiring long-term follow-up data that are, most of time, not available. The present analysis aims at characterizing the kinetics of antibodies against Porcine Reproductive and Respiratory Syndrome virus (PRRSv) from longitudinal data sets. The first data set consisted in the serological follow-up of 22 vaccinated gilts during 21 weeks post-vaccination (PV). The second one gathered the maternally derived antibodies (MDAs) kinetics in piglets from three different farms up to 14 weeks of age. The peak of the PV serological response against PRRSv was reached 6.9 weeks PV on average with an average duration of antibodies persistence of 26.5 weeks. In the monitored cohort of piglets, the duration of passive immunity was found relatively short, with an average duration of 4.8 weeks. The level of PRRSv-MDAs was found correlated with the dams’ antibody titer at birth, and the antibody persistence was strongly related to the initial MDAs titers in piglets. These results evidenced the importance of PRRSv vaccination schedule in sows, to optimize the delivery of antibodies to suckling piglets. These estimates of the duration of active and passive immunity could be further used as input parameters of epidemiological models to analyze their impact on the persistence of PRRSv within farms. PMID:29435455

  13. Evaluation of Control Strategies for Porcine Reproductive and Respiratory Syndrome (PRRS) in Swine Breeding Herds Using a Discrete Event Agent-Based Model

    Science.gov (United States)

    Arruda, Andréia Gonçalves; Friendship, Robert; Carpenter, Jane; Greer, Amy; Poljak, Zvonimir

    2016-01-01

    The objective of this study was to develop a discrete event agent-based stochastic model to explore the likelihood of the occurrence of porcine reproductive and respiratory syndrome (PRRS) outbreaks in swine herds with different PRRS control measures in place. The control measures evaluated included vaccination with a modified-live attenuated vaccine and live-virus inoculation of gilts, and both were compared to a baseline scenario where no control measures were in place. A typical North American 1,000-sow farrow-to-wean swine herd was used as a model, with production and disease parameters estimated from the literature and expert opinion. The model constructed herein was not only able to capture individual animal heterogeneity in immunity to and shedding of the PRRS virus, but also the dynamic animal flow and contact structure typical in such herds under field conditions. The model outcomes included maximum number of females infected per simulation, and time at which that happened and the incidence of infected weaned piglets during the first year of challenge-virus introduction. Results showed that the baseline scenario produced a larger percentage of simulations resulting in outbreaks compared to the control scenarios, and interestingly some of the outbreaks occurred over long periods after virus introduction. The live-virus inoculation scenario showed promising results, with fewer simulations resulting in outbreaks than the other scenarios, but the negative impacts of maintaining a PRRS-positive population should be considered. Finally, under the assumptions of the current model, neither of the control strategies prevented the infection from spreading to the piglet population, which highlights the importance of maintaining internal biosecurity practices at the farrowing room level. PMID:27875546

  14. [Correlation between antibodies against porcine reproductive and respiratory syndrome virus and pathological-anatomical organ findings in slaughter pigs at farm level].

    Science.gov (United States)

    Grünberger, B; Schleicher, C; Stüger, H-P; Reisp, K; Schmoll, F; Köfer, J; Sattler, T

    2015-01-01

    The porcine reproductive and respiratory syndrome (PRRS) worldwide causes important economic losses in pig production. Its causative agent, the PRRS virus (PRRSV), is one of the most frequently detected infectious agents in relation to respiratory diseases in pigs in Austria. We investigated the correlation between the PRRSV status of pig farms, determined by detection of PRRSV antibodies in the serum of slaughter pigs, and the prevalence of pathological-anatomical lung lesions in slaughter pigs of the respective farms. Between December 1, 2011 and April 16, 2012, a total of 1056 serum samples of slaughter pigs from 66 pig farms were collected at an Austrian abattoir. The presence of PRRSV antibodies was tested by enzyme-linked immunosorbent assay in each sample and the PRRSV status of the respective farms was determined. No PRRSV vaccination was performed on any of the farms. In addition, the pathological-anatomical lung lesions of all slaughter pigs of the 66 farms that were slaughtered between September 1, 2011 and December 11, 2012 were recorded by authorized veterinarians at the abattoir. The prevalence of lung lesions and pleuritis in PRRSV-positive and unsuspected farms was compared and statistically interpreted. Slaughter pigs of PRRSV positive farms had a significantly higher prevalence of severe lung lesions and pleuritis visceralis and parietalis than slaughter pigs of PRRSV unsuspected farms. Pigs of combined farms (nursery and fattening unit at the same location) displayed a tendency for more moderate and severe lung lesions than pigs of exclusive fattening farms. In the present study, the PRRSV status of pig farms displayed a significant influence on the prevalence of lung lesions in the slaughter pigs. Findings untypical for PRRS, including pleuritis, were also found significantly more often on those farms. This leads to the conclusion that other primary and/or secondary infections are involved, which can be exacerbated by the immunosuppressive

  15. Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain.

    Science.gov (United States)

    Wang, Qian; Li, Yanwei; Dong, Hong; Wang, Li; Peng, Jinmei; An, Tongqing; Yang, Xufu; Tian, Zhijun; Cai, Xuehui

    2017-02-22

    The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) continues to pose one of the greatest threats to the swine industry. M protein is the most conserved and important structural protein of PRRSV. However, information about the host cellular proteins that interact with M protein remains limited. Host cellular proteins that interact with the M protein of HP-PRRSV were immunoprecipitated from MARC-145 cells infected with PRRSV HuN4-F112 using the M monoclonal antibody (mAb). The differentially expressed proteins were identified by LC-MS/MS. The screened proteins were used for bioinformatics analysis including Gene Ontology, the interaction network, and the enriched KEGG pathways. Some interested cellular proteins were validated to interact with M protein by CO-IP. The PRRSV HuN4-F112 infection group had 10 bands compared with the control group. The bands included 219 non-redundant cellular proteins that interact with M protein, which were identified by LC-MS/MS with high confidence. The gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway bioinformatic analyses indicated that the identified proteins could be assigned to several different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with protein translation, infectious disease, and signal transduction. Two interested cellular proteins-nuclear factor of activated T cells 45 kDa (NF45) and proliferating cell nuclear antigen (PCNA)-that could interact with M protein were validated by Co-IP and confocal analyses. The interactome data between PRRSV M protein and cellular proteins were identified and contribute to the understanding of the roles of M protein in the replication and pathogenesis of PRRSV. The interactome of M protein will aid studies of virus/host interactions and provide means to decrease the threat of PRRSV to the swine industry in the future.

  16. Improved Cytotoxic T Lymphocyte Responses to Vaccination with Porcine Reproductive and Respiratory Syndrome Virus in 4-1BB Transgenic Pigs

    Directory of Open Access Journals (Sweden)

    Guangping Huang

    2017-12-01

    Full Text Available Vaccination is the most reliable measure to prevent infectious diseases in domestic animals. Development of novel vaccines demands extensive studies with new technologies, such as using novel adjuvants and immunomodulatory molecules. The co-stimulatory molecule 4-1BB provides a key signal that directs the fate of T cells during activation, and thus is important to their function in immune protection. To determine whether host immune responses to viral infection could be promoted by enhancing 4-1BB co-stimulation, in this study, we produced transgenic pig clones expressing an extra copy of the 4-1BB gene by clustered regularly interspaced short palindromic repeats/CRISPR-associated gene 9-mediated homologous recombination at the Rosa26 locus. The immune responses of transgenic pigs to porcine reproductive and respiratory syndrome virus (PRRSV vaccine were determined on day 14. We show that peripheral blood lymphocytes of transgenic pigs expressed around twice the level of 4-1BB mRNA than those of control pigs. We also found IL-2, TNF-α, and granzyme B mRNA levels as well as PRRSV-specific IFN-γ response were significantly upregulated in 4-1BB transgenic pigs, leading to more efficient cytotoxic T lymphocyte (CTL killing, whereas the expressions of IL-4, IL-17, and Foxp3 were not affected. These results indicate that higher levels of 4-1BB expression involve in promoting Th1 differentiation and enhancing specific CTL responses to PRRSV, and provide a novel approach to increase the efficacy of current vaccines to control the infectious diseases.

  17. reproduction

    African Journals Online (AJOL)

    progress in terms of increasing healthy live births but decreasing multiple pregnancy rates.10. Development of assisted reproduction techniques. Alternatives to IVF and transcervical embryo transfer. Over the years IVF treatment has seen many modifications, and other options have been introduced. Prepared sperm may be ...

  18. reproduction

    African Journals Online (AJOL)

    Examination ofHurnan Semen and Semen-Cervical Mucus. Interaction.20 Furthermore, organisations such as the. WHO and the European Society of Human Reproduction and Embryology (ESHRE) now set up international training courses aiming at global standardisation. Micromanipulation of gametes and male infertility.

  19. Serum resistance of Pasteurella multocida in avian and porcine sera, and comparative virulence investigations of selected serum-sensitive and resistant strains in chickens.

    Science.gov (United States)

    Muhairwa, Amandus P; Christensen, Jens P; Bisgaard, Magne

    2002-04-01

    Growth in serum of Pasteurella multocida and related species in chicken, turkey, duck and pig sera were compared, and selected serum-resistant and serum-sensitive strains were inoculated into 18-week-old layers. Eighty-seven field strains of Pasteurella spp. and nine reference strains representing different clones defined by restriction endonuclease analysis (REA) profiles were used in the study. Serum activity was measured by changes in the optical density (OD) of the serum after inoculation and incubation at 41 degrees C for chicken, turkey and duck serum and 39 degrees C for pig serum. Serum activity was measured by comparison with previously determined serum-resistant (P-1059) and serum-sensitive (CU vaccine) strains, and classified into highly serum-resistant, moderately serum-resistant and serum-sensitive. Strains of the same REA type were found to have identical growth curves and the same maximum OD values when tested in serum from the same host species. Turkey serum was shown to be less inhibitory to a wide range of P. multocida strains than chicken, duck and pig sera. Serum-resistant strains were demonstrated among avian as well as mammalian strains. Among the avian strains, the proportion of serum-resistant strains was higher in outbreak strains than in strains from apparently healthy carriers. Removal of the capsule from selected strains by hyaluronidase treatment failed to change the serum activity. The most severe lesions in experimentally infected chickens were produced by a serum-resistant strain; however, lesions were also found in chickens infected by serum-sensitive strains, indicating the involvement of multiple factors in the virulence of P. multocida. Further investigations on serum resistance are indicated in order to relate other host and bacterial factors responsible for the development of fowl cholera.

  20. A major gene for host response to porcine reproductive and respiratory syndrome is not unfavorably associated with overall performance under nonchallenging conditions in commercial pig lines.

    Science.gov (United States)

    Dunkelberger, J R; Mathur, P K; Lopes, M S; Knol, E F; Dekkers, J C M

    2017-07-01

    A QTL for host response to porcine reproductive and respiratory syndrome (PRRS) was identified in a previous study. The SNP WUR10000125 (WUR) is used as a tag SNP for this QTL. The favorable (B) allele at this SNP is in low frequency in commercial populations, possibly because this allele is unfavorably associated with an important trait under nonchallenging conditions and, therefore, may have been selected against. Therefore, objectives of this study were to estimate the effect of WUR on traits under selection in commercial lines under nonchallenging conditions and to estimate the effect of WUR genotype of parents on performance of crossbred progeny in a commercial-like environment. Data were collected on 4 purebred lines: a Landrace dam line (D1), a Large White dam line (D2), a synthetic sire line (S1), and a Pietrain sire line (S2). Traits analyzed included total number born, number stillborn, farrowing survival, lactation survival, litter mortality, daily feed intake (DFI), backfat, average test daily gain (TGR), average lifetime daily gain (LGR), and Topigs Norsvin selection index (TSI) value, indicative of overall economic value. Deregressed EBV were calculated for each trait (except TSI) and analyzed within line. In the S1 line, AB and BB pigs had significantly lower TGR ( = 0.002) and LGR ( = 0.001) than AA pigs but also lower DFI ( = 0.004). Conversely, AB and BB pigs had significantly higher DFI ( < 0.001) and AB pigs had significantly higher TGR ( = 0.03) than AA pigs in the S2 line. The effect of WUR on TSI was not significant for any line ( ≥ 0.15). Analyses of phenotypic records collected on crossbred progeny of S1 sires and D1 × D2 F females showed no significant effect of parent WUR genotype on DFI, backfat, TGR, or LGR ( ≥ 0.07). In conclusion, the effect of WUR was nonsignificant for most traits but the magnitude and direction of the effect differed by trait and by line. The favorable allele for host response to PRRS was associated with

  1. Genome-wide analysis of the transcriptional response to porcine reproductive and respiratory syndrome virus infection at the maternal/fetal interface and in the fetus.

    Science.gov (United States)

    Wilkinson, Jamie M; Bao, Hua; Ladinig, Andrea; Hong, Linjun; Stothard, Paul; Lunney, Joan K; Plastow, Graham S; Harding, John C S

    2016-05-20

    Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection of pregnant pigs can result in congenital infection and ultimately fetal death. Little is known about immune responses to infection at the maternal-fetal interface and in the fetus itself, or the molecular events behind virus transmission and disease progression in the fetus. To investigate these processes, RNA-sequencing of two sites, uterine endothelium with adherent placental tissue and fetal thymus, was performed 21 days post-challenge on four groups of fetuses selected from a large PRRSV challenge experiment of pregnant gilts: control (CON), uninfected (UNINF), infected (INF), and meconium-stained (MEC) (n = 12/group). Transcriptional analyses consisted of multiple contrasts between groups using two approaches: differential gene expression analysis and weighted gene co-expression network analysis (WGCNA). Biological functions, pathways, and regulators enriched for differentially expressed genes or module members were identified through functional annotation analyses. Expression data were validated by reverse transcription quantitative polymerase chain reaction (RTqPCR) carried out for 16 genes of interest. The immune response to infection in endometrium was mainly adaptive in nature, with the most upregulated genes functioning in either humoral or cell-mediated immunity. In contrast, the expression profile of infected fetal thymus revealed a predominantly innate immune response to infection, featuring the upregulation of genes regulated by type I interferon and pro-inflammatory cytokines. Fetal infection was associated with an increase in viral load coupled with a reduction in T cell signaling in the endometrium that could be due to PRRSV-controlled apoptosis of uninfected bystander cells. There was also evidence for a reduction in TWIST1 activity, a transcription factor involved in placental implantation and maturation, which could facilitate virus transmission or fetal pathology

  2. Measuring Progress on the Control of Porcine Reproductive and Respiratory Syndrome (PRRS at a Regional Level: The Minnesota N212 Regional Control Project (Rcp as a Working Example.

    Directory of Open Access Journals (Sweden)

    Pablo Valdes-Donoso

    Full Text Available Due to the highly transmissible nature of porcine reproductive and respiratory syndrome (PRRS, implementation of regional programs to control the disease may be critical. Because PRRS is not reported in the US, numerous voluntary regional control projects (RCPs have been established. However, the effect of RCPs on PRRS control has not been assessed yet. This study aims to quantify the extent to which RCPs contribute to PRRS control by proposing a methodological framework to evaluate the progress of RCPs. Information collected between July 2012 and June 2015 from the Minnesota Voluntary Regional PRRS Elimination Project (RCP-N212 was used. Demography of premises (e.g. composition of farms with sows = SS and without sows = NSS was assessed by a repeated analysis of variance. By using general linear mixed-effects models, active participation of farms enrolled in the RCP-N212, defined as the decision to share (or not to share PRRS status, was evaluated and used as a predictor, along with other variables, to assess the PRRS trend over time. Additionally, spatial and temporal patterns of farmers' participation and the disease dynamics were investigated. The number of farms enrolled in RCP-N212 and its geographical coverage increased, but the proportion of SS and NSS did not vary significantly over time. A significant increasing (p<0.001 trend in farmers' decision to share PRRS status was observed, but with NSS producers less willing to report and a large variability between counties. The incidence of PRRS significantly (p<0.001 decreased, showing a negative correlation between degree of participation and occurrence of PRRS (p<0.001 and a positive correlation with farm density at the county level (p = 0.02. Despite a noted decrease in PRRS, significant spatio-temporal patterns of incidence of the disease over 3-weeks and 3-kms during the entire study period were identified. This study established a systematic approach to quantify the effect of RCPs on

  3. Import risk assessment incorporating a dose-response model: introduction of highly pathogenic porcine reproductive and respiratory syndrome into Australia via illegally imported raw pork.

    Science.gov (United States)

    Brookes, V J; Hernández-Jover, M; Holyoake, P; Ward, M P

    2014-03-01

    Highly pathogenic porcine reproductive and respiratory syndrome (PRRS) has spread through parts of south-east Asia, posing a risk to Australia. The objective of this study was to assess the probability of infection of a feral or domestic pig in Australia with highly pathogenic PRRS following ingestion of illegally imported raw pork. A conservative scenario was considered in which 500 g of raw pork was imported from the Philippines into Australia without being detected by border security, then discarded from a household and potentially accessed by a pig. Monte Carlo simulation of a two-dimensional, stochastic model was used to estimate the probability of entry and exposure, and the probability of infection was assessed by incorporating a virus-decay and mechanistic dose-response model. Results indicated that the probability of infection of a feral pig after ingestion of raw meat was higher than the probability of infection of a domestic pig. Sensitivity analysis was used to assess the influence of input parameters on model output probability estimates, and extension of the virus-decay and dose-response model was used to explore the impact of different temperatures and time from slaughter to ingestion of the meat, different weights of meat, and the level of viraemia at slaughter on the infectivity of meat. Parameters with the highest influence on the model output were the level of viraemia of a pig prior to slaughter and the probability of access by a feral pig to food-waste discarded on property surrounding a household. Extension of the decay and dose-response model showed that small pieces of meat (10 g) from a highly pathogenic PRRS viraemic pig could contain enough virus to have a high probability of infection of a pig, and that routes to Australia by sea or air from all highly pathogenic PRRS virus endemic countries were of interest dependent on the temperature of the raw meat during transport. This study highlighted the importance of mitigation strategies such

  4. Honeybee (Apis mellifera Venom Reinforces Viral Clearance during the Early Stage of Infection with Porcine Reproductive and Respiratory Syndrome Virus through the Up-Regulation of Th1-Specific Immune Responses

    Directory of Open Access Journals (Sweden)

    Jin-A Lee

    2015-05-01

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS is a chronic and immunosuppressive viral disease that is responsible for substantial economic losses for the swine industry. Honeybee venom (HBV is known to possess several beneficial biological properties, particularly, immunomodulatory effects. Therefore, this study aimed at evaluating the effects of HBV on the immune response and viral clearance during the early stage of infection with porcine reproductive and respiratory syndrome virus (PRRSV in pigs. HBV was administered via three routes of nasal, neck, and rectal and then the pigs were inoculated with PRRSV intranasally. The CD4+/CD8+ cell ratio and levels of interferon (IFN-γ and interleukin (IL-12 were significantly increased in the HBV-administered healthy pigs via nasal and rectal administration. In experimentally PRRSV-challenged pigs with virus, the viral genome load in the serum, lung, bronchial lymph nodes and tonsil was significantly decreased, as was the severity of interstitial pneumonia, in the nasal and rectal administration group. Furthermore, the levels of Th1 cytokines (IFN-γ and IL-12 were significantly increased, along with up-regulation of pro-inflammatory cytokines (TNF-α and IL-1β with HBV administration. Thus, HBV administration—especially via the nasal or rectal route—could be a suitable strategy for immune enhancement and prevention of PRRSV infection in pigs.

  5. Generation of an infectious clone of VR-2332, a highly virulent North American type isolate of porcine reproductive and respiratory syndrome virus

    DEFF Research Database (Denmark)

    Nielsen, H.S.; Liu, G.; Nielsen, Jens

    2003-01-01

    -2332 strain. However, the cloned virus was clearly distinguishable from the parental VR-2332 strain by an engineered marker, a BstZ171 restriction site. The full-length cDNA clone had 11 nucleotide changes, 2 of which affected coding, compared to the parental VR-2332 strain. Additionally...... virus were consistently slightly lower. In experimentally infected 5.5-week-old pigs, the cloned virus produced blue discoloration of the ears, a classical clinical symptom of PRRSV. Also, the seroconversion kinetics of pigs infected with the cloned virus and VR-2332 were very similar. Hence, virus...

  6. Evaluation of protective immunity in gilts inoculated with the NADC-8 isolate of porcine reproductive and respiratory syndrome virus (PRRSV) and challenge-exposed with an antigenically distinct PRRSV isolate.

    Science.gov (United States)

    Lager, K M; Mengeling, W L; Brockmeier, S L

    1999-08-01

    To determine whether intrauterine inoculation of porcine reproductive and respiratory syndrome virus (PRRSV) interferes with conception and whether exposure to one strain of PRRSV provides protection against challenge-exposure (CE) with homologous or heterologous strains of PRRSV. 40 gilts. Gilts were inoculated by intrauterine administration of a PRRSV isolate (NADC-8) at breeding. Inoculated and noninoculated gilts were exposed oronasally to homologous (NADC-8) or heterologous (European isolate) PRRSV during late gestation. Specimens from gilts and fetuses were tested against CE virus. Lack of virus in gilts indicated protective immunity for the dam, in fetuses indicated protection of gilt from reproductive losses, and in both groups indicated complete protection. In the homologous CE group, interval from inoculation to CE ranged from 90 to 205 days, and protection was complete. In the heterologous CE group, interval from inoculation to CE ranged from 90 to 170 days, and protection was incomplete. The CE virus was detected in gilts necropsied 134 to 170 days after CE and in a litter necropsied 170 days after CE. Homologous protection can be induced in gilts by exposure to live PRRSV. Heterologous protection from reproductive losses can be induced in gilts by exposure to live PRRSV; however, this protection is incomplete and may have a shorter duration than homologous protection. Exposure of swine to enzootic PRRSV will provide protection against homologous PRRSV-induced reproductive losses. Extent and duration of protection against heterologous PRRSV may be variable and dependent on antigenic relatedness of the virus strains used for inoculation and CE.

  7. Reproduction of postweaning multisystemic wasting syndrome (PMWS) in immunostimulated and non-immunostimulated 3-week-old piglets experimentally infected with porcine circovirus type 2 (PCV2)

    DEFF Research Database (Denmark)

    Ladekjær-Mikkelsen, A.S.; Nielsen, Jens; Stadejek, T.

    2002-01-01

    Postweaning multisystemic wasting syndrome (PMWS) in swine is causally associated with the newly recognised pathogen, porcine circovirus type 2 (PCV2). In this study, 3-week-old SPF PCV2-seronegative piglets were inoculated intranasally with PCV2. The effect of immunostimulation on the induction...

  8. Dietary Probiotic Compound Improves Reproductive Performance of Porcine Epidemic Diarrhea Virus-Infected Sows Reared in a Japanese Commercial Swine Farm under Vaccine Control Condition

    Directory of Open Access Journals (Sweden)

    Takio Inatomi

    2017-12-01

    Full Text Available Lactogenic immunity transferred to piglets after inoculation of a live vaccine to pregnant sows was proved limited to control porcine epidemic diarrhea (PED. Hence, here we evaluated the efficacy of administration of a probiotic compound containing Bacillus mesentericus, Clostridium butyricum, and Enterococcus faecalis together with a commercial live-attenuated PED vaccine (Nisseiken PED Live Vaccine, Nisseiken, Tokyo, Japan to improve the health and reproductive performance of PED-infected sows. Twenty pregnant sows in a PED-positive farm were equally divided into probiotics-administered (VP and control (VC sow groups. A commercial live-attenuated vaccine was injected as per the manufacturer’s instruction. The probiotic compound (15 g/day was orally administered to VP from 6 weeks pre-parturition to 7 days post-parturition (ppd7. VP had a significantly higher body weight at ppd7 than VC (191 vs 186 kg; P < 0.05. At day 3 post-parturition (ppd3 (4.18 vs 3.63 kg/day and ppd7 (5.14 vs 4.34 kg/day, milk produced by VP was significantly (P < 0.05 greater than that by VC. Total immunoglobulin (IgA and IgG concentrations at day 0 were significantly (P < 0.05 higher in whey of VP (1.9 and 6.6 g/dL, respectively than in that of VC (1.7 and 6.1 g/dL, respectively. However, total IgG concentration in whey of VP and VC at ppd3 and ppd7 did not differ. Antibody titer was significantly higher at day 0 in serum of VP than it was that of VC (60 vs 37 in geometric mean; P < 0.05. Likewise, the antibody titer in whey of VP and VC was found to be similar at day 0 (416 vs 208 in geometric mean; P = 0.13. Consequently, VP had fewer days between weaning and return to estrus than did VC (7 vs 10 days; P < 0.05. Moreover, piglets of VP had a significantly (P < 0.05 higher litter weight at birth (9,252 g/litter and a lower mortality (12% during suckling than those of VC (8,686 g/litter and 28%, respectively. In

  9. In utero infection with porcine reproductive and respiratory syndrome virus modulates leukocyte subpopulations in peripheral blood and bronchoalveolar fluid of surviving piglets.

    Science.gov (United States)

    Nielsen, J; Bøtner, A; Tingstedt, J-E; Aasted, B; Johnsen, C K; Riber, U; Lind, P

    2003-06-20

    It is well known that piglets congenitally infected with porcine reproductive and respiratory syndrome virus (PRRSV) can be viremic at birth, and that preweaning mortality due to secondary infections often increases during acute outbreaks of PRRS. Therefore, an immunosuppressive effect of in utero infection has been suggested. The aim of the present study was to characterise the changes of leukocyte populations in piglets surviving in utero infection with PRRSV. A total of 27 liveborn uninfected control piglets and 22 piglets infected transplacentally with a Danish strain of PRRSV were included. At 2 and 4 weeks of age, 21 of 22 (96%) and 7 of 14 (50%) examined infected piglets were still viremic, whereas PRRSV could not be detected in the six infected piglets examined at 6 weeks of age. Flow cytometry analysis was used to determine the phenotypic composition of leukocytes in peripheral blood and bronchoalveolar lavage fluid (BALF) of 2-, 4- and 6-week-old infected piglets and age-matched uninfected controls. The key observation in the present study is that high levels of CD8(+) cells constitute a dominant feature in peripheral blood and BALF of piglets surviving in utero infection with PRRSV. In BALF, the average high level of CD8(+) cells in 2-week-old infected piglets (33.4 +/- 12.6%) was followed by a decline to 7.3 +/- 3.0 and 11.1 +/- 3.0% at 4 and 6 weeks of age. BALF of control piglets contained 1.6 +/- 0.9, 2.3 +/- 1.8 and 1.9 +/- 0.5% CD8(+) cells, only. In peripheral blood, however, the average number of CD8(+) cells remained at high levels in the infected piglets throughout the post-natal experimental period (2.8 +/- 1.9, 2.9 +/- 1.8 and 3.2 +/- 1.7 x 10(6) CD8(+) cells/ml at 2, 4 and 6 weeks, respectively). In the controls, the average levels of CD8(+) cells were 0.9+/-0.2, 1.9 +/- 1.7 and 1.6 +/- 0.5 x 10(6)/ml, respectively. Furthermore, the numbers of CD2(+) , CD4(+)CD8(+) and SLA-classII(+) cells, respectively, in peripheral blood, together with

  10. 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

    Directory of Open Access Journals (Sweden)

    William A. Cafruny

    2008-01-01

    Full Text Available 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 quinolonecontaining compound Plasmocin™, as well as the quinolones nalidixic acid and ciprofloxacin, have potent anti-PRRSV activity in vitro. PRRSV replication was inhibited by these antibiotics in both cultured MARC-145 cells and cultured primary alveolar porcine macrophages (PAMs. Furthermore, sub-optimal concentrations of nalidixic acid synergized with antiviral cytokines (AK-2 or IFN-γ to quantitatively and qualitatively inhibit PRRSV replication in MARC-145 cells or PAMs. The antiviral activity of Plasmocin and nalidixic acid correlated with reduced actin expression in MARC-145 cells. Replication of the related lactate dehydrogenase-elevating virus (LDV was also inhibited in primary mouse macrophages by Plasmocin. These results are significant to the development of antiviral strategies with potentially reduced toxicity, and provide a model system to better understand regulation of arterivirus replication.

  11. Evaluation of the effectiveness of an antimicrobial air filter to avoid porcine reproductive and respiratory syndrome virus (PRRSV) aerosol transmission, after 16 months of exposure to a commercial swine environmental conditions

    Energy Technology Data Exchange (ETDEWEB)

    Batista, L. [Boehringer Ingelheim Vetmedica Inc., St. Joseph, MO (United States); Pouliot, F.; Urizar, L. [Centre de developpement du porc du Quebec Inc., Quebec City, PQ (Canada)

    2010-07-01

    The effectiveness of Noveko's antimicrobial filter was evaluated after 16 months of exposure to commercial swine production. The experiment involved the use of a scaled model of a commercial swine facility consisting of 2 small chambers connected by a duct containing the filters. A 5 kg naive pig was placed in the reception chamber for a period of 6 hours after aerosolization with porcine reproductive and respiratory syndrome virus (PRRSV). Blood samples from pigs were collected before and after aerosolization to test for the presence of PRRSV RNA. Only blood samples were tested for PRRSV antibodies by IDEXX 2XR ELISA. None of the 9 pigs tested were found to be infected. The study showed that the technology used to integrate the antimicrobial agent into the filter fibers allows the filter combination to withstand extreme weather and endure commercial swine production for at least 16 months, and can maintain its effectiveness to avoid airborne transmission of PRRSV.

  12. Robust Protection against Highly Virulent Foot-and-Mouth Disease Virus in Swine by Combination Treatment with Recombinant Adenoviruses Expressing Porcine Alpha and Gamma Interferons and Multiple Small Interfering RNAs

    Science.gov (United States)

    Park, Jong-Hyeon; Lee, Kwang-Nyeong; Kim, Se-Kyung; You, Su-Hwa; Kim, Taeseong; Tark, Dongseob; Lee, Hyang-Sim; Seo, Min-Goo; Kim, Byounghan

    2015-01-01

    ABSTRACT Because the currently available vaccines against foot-and-mouth disease (FMD) provide no protection until 4 to 7 days postvaccination, the only alternative method to halt the spread of the FMD virus (FMDV) during outbreaks is the application of antiviral agents. Combination treatment strategies have been used to enhance the efficacy of antiviral agents, and such strategies may be advantageous in overcoming viral mechanisms of resistance to antiviral treatments. We have developed recombinant adenoviruses (Ads) for the simultaneous expression of porcine alpha and gamma interferons (Ad-porcine IFN-αγ) as well as 3 small interfering RNAs (Ad-3siRNA) targeting FMDV mRNAs encoding nonstructural proteins. The antiviral effects of Ad-porcine IFN-αγ and Ad-3siRNA expression were tested in combination in porcine cells, suckling mice, and swine. We observed enhanced antiviral effects in porcine cells and mice as well as robust protection against the highly pathogenic strain O/Andong/SKR/2010 and increased expression of cytokines in swine following combination treatment. In addition, we showed that combination treatment was effective against all serotypes of FMDV. Therefore, we suggest that the combined treatment with Ad-porcine IFN-αγ and Ad-3siRNA may offer fast-acting antiviral protection and be used with a vaccine during the period that the vaccine does not provide protection against FMD. IMPORTANCE The use of current foot-and-mouth disease (FMD) vaccines to induce rapid protection provides limited effectiveness because the protection does not become effective until a minimum of 4 days after vaccination. Therefore, during outbreaks antiviral agents remain the only available treatment to confer rapid protection and reduce the spread of foot-and-mouth disease virus (FMDV) in livestock until vaccine-induced protective immunity can become effective. Interferons (IFNs) and small interfering RNAs (siRNAs) have been reported to be effective antiviral agents against

  13. Colostral transmission of porcine circovirus 2 (PCV-2): reproduction of post-weaning multisystemic wasting syndrome in pigs fed milk from PCV-2-infected sows with post-natal porcine parvovirus infection or immunostimulation.

    Science.gov (United States)

    Ha, Yooncheol; Shin, Jeoung Hwa; Chae, Chanhee

    2010-06-01

    Post-weaning multisystemic wasting syndrome (PMWS) was reproduced in pigs fed colostrum and milk from porcine circovirus 2 (PCV-2)-infected sows and infected post-natally with porcine parvovirus (PPV) or immunostimulated. Pregnant sows were inoculated intranasally with either PCV-2 (n=5) or PCV-2-free PK-15 cell lysates (control, n=10) 3 weeks before the expected farrowing date. Newborn piglets from five of the control sows were introduced to PCV-2-infected sows (n=6 for each sow) and allowed to feed on the colostrum for 12 h and then given 15 ml milk five times a day for 7 days. Newborn piglets from the other five control sows were fed colostrum and milk from their own sows. After 7 days, two piglets from each group were randomly selected to confirm PCV-2 infection. Twenty-one pigs fed by PCV-2-infected sows were randomly divided into three groups and subjected to post-natal PPV infection (group 1), immunostimulation (group 2) or no post-natal treatment (group 3). Twenty-one pigs fed by uninfected sows were also randomly divided and subjected to post-natal PCV-2 and PPV infection (group 4), post-natal PCV-2 infection (group 5) or no treatment (group 6, negative control). Body weight was significantly greater in group 6 than in groups 1, 2 and 4 at 49, 52, 56, 59 and 63 days of age. The typical granulomatous inflammatory reaction and lymphoid depletion of PMWS was observed in the lymph nodes of groups 1, 2 and 4 at 63 days of age. Group 3 had significantly fewer PCV-2-positive cells than groups 1, 2 and 4. In conclusion, PCV-2 shed from colostrum and milk is infectious and reproduces PMWS with post-natal PPV infection or immune stimulation.

  14. Porcine eperythrozoonosis in China.

    Science.gov (United States)

    Wu, Jiansan; Yu, Jianmin; Song, Cuiping; Sun, Shengjun; Wang, Zhiliang

    2006-10-01

    Eperythrozoonosis of swine (also designated as porcine mycoplasmosis) is a disease of swine under stress, expressed as a febrile condition with development of an acute ictero-anemia. It is caused by Eperythrozoon suis and usually causes a subclinical infection with a latent carrier state that persists for extended periods. In China, this disease has gradually developed as an important intercurrent disease and an emerging swine disease that, in recent years, has spread throughout all provinces except Tibet. Classical swine fever (hog cholera), porcine influenza, swine enzootic pneumonia, porcine reproductive and respiratory syndrome (blue ear disease), streptococci, and toxoplasmosis were detected in Eperythrozoonosis-infected pig herds, and caused serious economic losses. National epidemiology surveillance in 2002 revealed that this disease caused a total morbidity of 30% and a mortality of 10-20%. Total mortality (which includes culling sick pigs) was more than 60%. The morbidity within infected herds was near 100%, has spread throughout with a total mortality rate usually over 50%. Mortality of piglets in some districts was as high as 50%. The highest infection rate on pig farms was more than 90%. The farms with higher infection rates occurred in pig-raising areas during epidemic seasons. New diagnostic tests, such as ELISA and PCR, have been developed for the detection of porcine eperythrozoonosis, but traditionally the diagnosis of the disease is still based on clinical history and optical microscopic examination of the causative agent in blood smears. Efficient preventive and control measures include the detection of carriers in pig herds and treatment of sick pigs with drugs, such as long-acting oxytetracycline, doxycycline, or aceturate of diminazene. Oxytetracyclines as feed additives have been introduced for eperythrozoonosis prevention in uninfected pig herds, and pig producers have taken measures to reduce stress and improve sanitary conditions.

  15. Plasmids expressing interleukin-10 short hairpin RNA mediate IL-10 knockdown and enhance tumor necrosis factor alpha and interferon gamma expressions in response to porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Charerntantanakul, Wasin; Kasinrerk, Watchara

    2012-04-15

    Porcine reproductive and respiratory syndrome virus (PRRSV) has been suggested to exploit interleukin-10 (IL-10) to suppress immune defense of infected pigs. The present study constructed plasmids encoding selected short hairpin RNA specific to porcine IL-10 mRNA (pIL-10sh) to knockdown IL-10 transcription and investigated the suppressive effect of PRRSV-induced IL-10 on various immune marker expressions. Naïve blood monocytes from eight PRRSV-seronegative pigs were transfected with pIL-10sh and pNeg (plasmid vector) prior to PRRSV inoculation and subsequent lipopolysaccharide (LPS) stimulation. The mRNA expressions of IL-10, IL-1β, IL-12p40, tumor necrosis factor alpha (TNFα), interferon gamma (IFNγ), transforming growth factor beta (TGFβ), CD80, and CD86 were evaluated by real-time PCR. The IL-10, TNFα, and IFNγ protein productions were determined by ELISA. Compared with non-transfected monocyte control, transfection with selected pIL-10sh (pIL-10sh1), but not other pIL-10sh nor pNeg, significantly reduced IL-10 expression and significantly enhanced TNFα and IFNγ expressions. Slight increases in IL-1β, IL-12p40, CD80, and CD86 expressions were also observed. Neither pIL-10sh1 nor pNeg transfection affected TGFβ expression. Our results indicate that PRRSV does exploit IL-10 to suppress the expressions of pro-inflammatory cytokines, mainly TNFα and IFNγ, and co-stimulatory molecules, CD80 and CD86. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Variation in Fetal Outcome, Viral Load and ORF5 Sequence Mutations in a Large Scale Study of Phenotypic Responses to Late Gestation Exposure to Type 2 Porcine Reproductive and Respiratory Syndrome Virus

    Science.gov (United States)

    Ladinig, Andrea; Wilkinson, Jamie; Ashley, Carolyn; Detmer, Susan E.; Lunney, Joan K.; Plastow, Graham; Harding, John C. S.

    2014-01-01

    In spite of extensive research, the mechanisms of reproductive disease associated with Porcine Reproductive and Respiratory Syndrome virus (PRRSv) are still poorly understood. The objectives of this large scale study were to evaluate associations between viral load and fetal preservation, determine the impact of type 2 PRRSv on fetal weights, and investigate changes in ORF5 PRRSv genome in dams and fetuses during a 21-day period following challenge. At gestation day 85 (±1), 114 gilts were experimentally infected with type 2 PRRSv, while 19 gilts served as reference controls. At necropsy, fetuses were categorized according to their preservation status and tissue samples were collected. PRRSv RNA concentrations were measured in gilt serum collected on days 0, 2, 6, and 21 post-infection, as well as in gilt and fetal tissues collected at termination. Fetal mortality was 41±22.8% in PRRS infected litters. Dead fetuses appeared to cluster in some litters but appeared solitary or random in others. Nine percent of surviving piglets were meconium-stained. PRRSv RNA concentration in fetal thymus, fetal serum and endometrium differed significantly across preservation category and was greatest in tissues of meconium-stained fetuses. This, together with the virtual absence of meconium staining in non-infected litters indicates it is an early pathological condition of reproductive PRRS. Viral load in fetal thymus and in fetal serum was positively associated with viral load in endometrium, suggesting the virus exploits dynamic linkages between individual maternal-fetal compartments. Point mutations in ORF5 sequences from gilts and fetuses were randomly located in 20 positions in ORF5, but neither nucleotide nor amino acid substitutions were associated with fetal preservation. PRRSv infection decreased the weights of viable fetuses by approximately 17%. The considerable variation in gilt and fetal outcomes provides tremendous opportunity for more detailed investigations of

  17. Complete Genome Sequence of the Porcine Isolate Enterococcus faecalis D32

    DEFF Research Database (Denmark)

    Zischka, Melanie; Kuenne, Carsten; Blom, Jochen

    2012-01-01

    The complete and annotated genome sequence of Enterococcus faecalis D32, a commensal strain isolated from a Danish pig, suggests putative adaptation to the porcine host and absence of distinct virulence-associated traits.......The complete and annotated genome sequence of Enterococcus faecalis D32, a commensal strain isolated from a Danish pig, suggests putative adaptation to the porcine host and absence of distinct virulence-associated traits....

  18. Effect of porcine reproductive and respiratory syndrome virus (PRRSV) (isolate ATCC VR-2385) infection on bactericidal activity of porcine pulmonary intravascular macrophages (PIMs): in vitro comparisons with pulmonary alveolar macrophages (PAMs).

    Science.gov (United States)

    Thanawongnuwech, R; Thacker, E L; Halbur, P G

    1997-11-01

    Porcine pulmonary intravascular macrophages (PIMs) were recovered by in situ pulmonary vascular perfusion with 0.025% collagenase in saline from six 8-week old, crossbred pigs. Pulmonary alveolar macrophages (PAMs) were recovered by bronchoalveolar lavage from the same pigs for comparisons in each assay. The macrophages were exposed to PRRSV (ATCC VR-2385) in vitro for 24 h and infection was confirmed by an indirect immunofluorescence test or transmission electron microscopy. Viral particles tended to accumulate in the vesicles of the Golgi apparatus or endoplasmic reticulum. Bactericidal function assays were performed on the recovered macrophages to determine the effects of the virus on macrophage functions. In vitro PRRSV infection reduced the bactericidal ability of PIMs from 68.3% to 56.4% (P 0.1) at 24 h post-infection. The mean percentage of bacteria killed by macrophages after PRRSV infection was not significantly different among the treatment groups or between the treatment groups and non-infected controls based on colorimetric MTT bactericidal (Staphylococcus aureus) assay. PRRSV did not affect the ability of PIMs or PAMs to internalize opsonized 125I-iododeoxyuridine-labeled S. aureus (P > 0.05). PRRSV infection significantly decreased the production of superoxide anion (P PIMs and by 69.4% in PAMs. PRRSV reduced the myeloperoxidase-H2O2-halide product (P PIMs and by 48.1% for PAMs. The results suggest: (1) PIMs should be considered as an important replication site of PRRSV; (2) PRRSV may have a detrimental effect on both PIMs and PAMs; (3) loss of bactericidal function in PIMs may facilitate hematogenous bacterial infections.

  19. Characterization of the Interactome of the Porcine Reproductive and Respiratory Syndrome Virus Nonstructural Protein 2 Reveals the Hyper Variable Region as a Binding Platform for Association with 14-3-3 Proteins.

    Science.gov (United States)

    Xiao, Yihong; Wu, Weining; Gao, Jiming; Smith, Nikki; Burkard, Christine; Xia, Dong; Zhang, Minxia; Wang, Chengbao; Archibald, Alan; Digard, Paul; Zhou, En-Min; Hiscox, Julian A

    2016-05-06

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry worldwide and hence global food security, exacerbated by a newly emerged highly pathogenic (HP-PRRSV) strain from China. PRRSV nonstructural protein 2 (nsp2) is a multifunctional polypeptide with strain-dependent influences on pathogenicity. A number of discrete functional regions have been identified on the protein. Quantitative label free proteomics was used to identify cellular binding partners of nsp2 expressed by HP-PRRSV. This allowed the identification of potential cellular interacting partners and the discrimination of nonspecific interactions. The interactome data were further investigated and validated using biological replicates and also compared with nsp2 from a low pathogenic (LP) strain of PRRSV. Validation included both forward and reverse pulldowns and confocal microscopy. The data indicated that nsp2 interacted with a number of cellular proteins including 14-3-3, CD2AP, and other components of cellular aggresomes. The hyper-variable region of nsp2 protein was identified as a binding platform for association with 14-3-3 proteins.

  20. A Review of Quantitative Tools Used to Assess the Epidemiology of Porcine Reproductive and Respiratory Syndrome in U.S. Swine Farms Using Dr. Morrison's Swine Health Monitoring Program Data.

    Science.gov (United States)

    Vilalta, Carles; Arruda, Andreia G; Tousignant, Steven J P; Valdes-Donoso, Pablo; Muellner, Petra; Muellner, Ulrich; Alkhamis, Moh A; Morrison, Robert B; Perez, Andres M

    2017-01-01

    Porcine reproductive and respiratory syndrome (PRRS) causes far-reaching financial losses to infected countries and regions, including the U.S. The Dr. Morrison's Swine Health Monitoring Program (MSHMP) is a voluntary initiative in which producers and veterinarians share sow farm PRRS status weekly to contribute to the understanding, in quantitative terms, of PRRS epidemiological dynamics and, ultimately, to support its control in the U.S. Here, we offer a review of a variety of analytic tools that were applied to MSHMP data to assess disease dynamics in quantitative terms to support the decision-making process for veterinarians and producers. Use of those methods has helped the U.S. swine industry to quantify the cyclical patterns of PRRS, to describe the impact that emerging pathogens has had on that pattern, to identify the nature and extent at which environmental factors (e.g., precipitation or land cover) influence PRRS risk, to identify PRRS virus emerging strains, and to assess the influence that voluntary reporting has on disease control. Results from the numerous studies reviewed here provide important insights into PRRS epidemiology that help to create the foundations for a near real-time prediction of disease risk, and, ultimately, will contribute to support the prevention and control of, arguably, one of the most devastating diseases affecting the North American swine industry. The review also demonstrates how different approaches to analyze and visualize the data may help to add value to the routine collection of surveillance data and support infectious animal disease control.

  1. Evaluation of monoclonal antibody-based immunohistochemistry for the detection of European and North American Porcine reproductive and respiratory syndrome virus and a comparison with in situ hybridization and reverse transcription polymerase chain reaction.

    Science.gov (United States)

    Han, Kiwon; Seo, Hwi Won; Oh, Yeonsu; Kang, Ikjae; Park, Changhoon; Kang, Sang Hoon; Kim, Sung-Hoon; Lee, Bog-Hieu; Kwon, Byungjoon; Chae, Chanhee

    2012-07-01

    The objective of the present study was to compare the ability of 2 monoclonal antibodies (mAbs; SDOW17 and SR30) to detect types 1 and 2 Porcine reproductive and respiratory syndrome virus (PRRSV) in formalin-fixed, paraffin-embedded (FFPE) lung tissues by immunohistochemistry (IHC) and to compare the immunohistochemical results with in situ hybridization (ISH) and reverse transcription nested polymerase chain reaction (RT-nPCR) detection techniques. Lungs from 30 experimentally infected pigs (15 pigs with each genotype of PRRSV) and 20 naturally infected pigs (10 pigs with each genotype of PRRSV) with types 1 and 2 PRRSV, respectively, were used for the IHC, ISH, and RT-nPCR analyses. The SR30 mAb-based IHC detected significantly more type 1 PRRSV-positive cells in the accessory and caudal lobes from the experimentally infected pigs at 7 (P = 0.025) and 14 (P = 0.018) days postinoculation, respectively, compared to the SDOW17 mAb-based IHC. The results demonstrated that SR30 mAb-based IHC is useful for detecting both types 1 and 2 PRRSV antigen in FFPE lung tissues.

  2. DExD/H-Box Helicase 36 Signaling via Myeloid Differentiation Primary Response Gene 88 Contributes to NF-κB Activation to Type 2 Porcine Reproductive and Respiratory Syndrome Virus Infection

    Directory of Open Access Journals (Sweden)

    Huiyuan Jing

    2017-10-01

    Full Text Available DExD/H-box helicase 36 (DHX36 is known to be an ATP-dependent RNA helicase that unwinds the guanine-quadruplexes DNA or RNA, but emerging data suggest that it also functions as pattern recognition receptor in innate immunity. Porcine reproductive and respiratory syndrome virus (PRRSV is an Arterivirus that has been devastating the swine industry worldwide. Interstitial pneumonia is considered to be one of the most obvious clinical signs of PRRSV infection, suggesting that the inflammatory response plays an important role in PRRSV pathogenesis. However, whether DHX36 is involved in PRRSV-induced inflammatory cytokine expression remains unclear. In this study, we found that PRRSV infection increased the expression of DHX36. Knockdown of DHX36 and its adaptor myeloid differentiation primary response gene 88 (MyD88 by small-interfering RNA in MARC-145 cells significantly reduced NF-κB activation and pro-inflammatory cytokine expression after PRRSV infection. Further investigation revealed that PRRSV nucleocapsid protein interacted with the N-terminal quadruplex binding domain of DHX36, which in turn augmented nucleocapsid protein-induced NF-κB activation. Taken together, our results suggest that DHX36–MyD88 has a relevant role in the recognition of PRRSV nucleocapsid protein and in the subsequent activation of pro-inflammatory NF-κB pathway.

  3. Cytokine mRNA profiles in bronchoalveolar cells of piglets experimentally infected in utero with porcine reproductive and respiratory syndrome virus: Association of sustained expression of IFN-gamma and IL-10 after viral clearance

    DEFF Research Database (Denmark)

    Johnsen, C. K.; Bøtner, Anette; Kamstrup, Søren

    2002-01-01

    An experimental model was used to investigate mRNA cytokine profiles in bronchoalvolar cells (BALC) from piglets, infected in utero with porcine reproductive and respiratory syndrome virus (PRRSV). The BALC's were analyzed for the cytokines TNF-alpha, IFN-gamma, IL-8, IL-10, and IL-12(p40) by real......-time TaqMan polymerase chain reaction in 2-, 4-, and 6-week-old piglets, respectively. High levels of IFN-gamma mRNA was detected in all piglets, while IL-10 was upregulated in 2-week-old piglets, was at normal levels in 4-week-old piglets, and elevated again in 6-week-old piglets. IL-12 was weakly...... elevated in all three age groups. Virus was reduced by 50% in 4-week-old piglets and cleared by 6 weeks of age. The sustained expression of IFNgamma and reduction of IL-10 production indicate an important role for these cytokines in immunity to PRRSV....

  4. Influence of pig age on virus titer and bactericidal activity of porcine reproductive and respiratory syndrome virus (PRRSV)-infected pulmonary intravascular macrophages (PIMs).

    Science.gov (United States)

    Thanawongnuwech, R; Thacker, E L; Halbur, P G

    1998-10-01

    Twelve pigs (six 4-week-old and six 4-month-old cross-bred, specific pathogen free pigs) were used as donors for both pulmonary intravascular macrophages (PIMs) and pulmonary alveolar macrophages (PAMs). The PIMs and PAMs were infected in vitro with low (ISU-55) or high (VR-2385) virulence strains of PRRSV at 1 multiplicity of infection (m.o.i.) for comparisons of virus titers at 48 h post infection (PI). PIMs were as permissive as PAMs to infection with both PRRSV isolates yielding similar progeny titers (10(4.81) vs. 10(5.22) TCID50/ml, respectively). Both ISU-55 and VR-2385 were able to infect PIMs and no significant difference in virus replication as measured by virus titers between isolates was found (10(5.33) vs. 10(4.69) TCID50/ml, respectively). PIMs from 4-weak-old pigs yielded a higher virus titer following PRRSV infection than PIMs from 4-month-old pigs (10(5.43) vs. 10(4.59) TCID50/ml, respectively; p PIMs had significantly decreased bactericidal (Staphylococcus aureus) activity compared with uninfected PIMS at 48 h PI (p PIMs and VR-2385 (high virulence)-infected PIMs. Both ISU-55 and VR-2385 infection significantly decreased the production of superoxide anion (SOA) at 24 and 48 h PI (p PIMs, (2) PIMs from younger pigs were more permissive to PRRSV infection, and (3) the selected PRRSV strains, which differ in their abilities to induce pneumonia in vivo were not different when tested in vitro by measuring virus titer and bactericidal functions.

  5. Stress-activated protein kinases are involved in porcine reproductive and respiratory syndrome virus infection and modulate virus-induced cytokine production.

    Science.gov (United States)

    Lee, Yoo Jin; Lee, Changhee

    2012-06-05

    The present study examined the role of the p38 MAPK and JNK pathways during PRRSV infection in immortalized porcine alveolar macrophage (PAM) cells. Infection with PRRSV was found to progressively activate p38 and JNK1/2 up to 36 h postinfection and then their phosphorylation levels dramatically decreased to baseline at 48 h postinfection. In contrast, UV-inactivated PRRSV failed to trigger phosphorylation of these SAPKs, indicating that the post-entry process is responsible for their activation. Independent treatment of cells with a selective p38 or JNK inhibitor markedly impaired PRRSV infection, resulting in significant reduction in synthesis of viral genomic and subgenomic RNAs, viral protein expression, and progeny virus production. Notably, cytokine production in PAM cells infected with PRRSV was shown to be altered by inhibiting these SAPKs. Altogether, our data suggest that the p38 and JNK signaling pathways play pivotal roles in PRRSV replication and may regulate immune responses during virus infection. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Isolation of porcine reproductive and respiratory syndrome (PRRS) virus in a Danish swine herd and experimental infection of pregnant gilts with the virus

    DEFF Research Database (Denmark)

    Bøtner, Anette; Nielsen, Jens; Bille-Hansen, Vivi

    1994-01-01

    , the virus particle was found to be spherical and enveloped, measuring 45–55 nm in diameter and containing a 30–35 nm nucleocapsid. Only minor antigenic differences were found between the Danish and a Dutch isolate. Following intranasal inoculation of 3 pregnant gilts with the Danish isolate transplacental...... infection was demonstrated by the re-isolation of PRRS virus from approximately 45% of the piglets from the experimentally infected gilts. However, the experimental infection produced no significant reproductive disorders or other clinical signs. At autopsy, histopathological examination revealed slight...

  7. Experimental reproduction of porcine circovirus type 2 (PCV2)-associated enteritis in pigs infected with PCV2 alone or concurrently with Lawsonia intracellularis or Salmonella typhimurium.

    Science.gov (United States)

    Opriessnig, T; Madson, D M; Roof, M; Layton, S M; Ramamoorthy, S; Meng, X J; Halbur, P G

    2011-01-01

    Porcine circovirus (PCV)-associated disease (PCVAD) has emerged to become one of the most economically important pig diseases globally. One of the less commonly recognized clinical manifestations of PCVAD is PCV2 type 2 (PCV2)-associated enteritis in growing pigs; however, experimental confirmation of the ability of PCV2 alone or PCV2 coinfection with other agent(s) to induce enteritis is lacking. In this study, 120 specific-pathogen-free (SPF) pigs were divided randomly into six groups: controls (negative control pigs), PCV2 (inoculated with PCV2), LAW (inoculated with Lawsonia intracellularis), SALM (inoculated with Salmonella typhimurium), PCV2-LAW (concurrently inoculated with PCV2 and Lawsonia intracellularis) and PCV2-SALM (concurrently inoculated with PCV2 and Salmonella typhimurium). One half of the pigs in each group were subject to necropsy examination 14 days postinoculation (dpi) and the remaining pigs were examined at 28 dpi. The average daily weight gain was not different (P>0.05) between groups. Individual pigs inoculated orally with PCV2 regardless of coinfection status (2/10 PCV2, 1/10 PCV2-LAW, 3/10 PCV2-SALM) developed PCVAD with diarrhoea and reduced weight gain or weight loss between 14 and 28 dpi. Those pigs had characteristic microscopic lesions in lymphoid and enteric tissues associated with abundant PCV2 antigen. Enteric lesions were characterized by necrosuppurative and proliferative enteritis with crypt elongation and epithelial hyperplasia in LAW and PCV2-LAW pigs by 14 dpi, ulcerative and necrosuppurative colitis in SALM and PCV2-SALM pigs by 14 dpi, and lymphohistiocytic enteritis with depletion of Peyer's patches in PCV2, PCV2-SALM and PCV2-LAW pigs by 28 dpi. To the authors' knowledge, this is the first report documenting that under experimental conditions, PCV2 can induce enteritis independently from other enteric pathogens and that oral challenge is a potentially important route and perhaps the natural route of PCV2 transmission in

  8. Association of porcine heparanase and hyaluronidase 1 and 2 with reproductive and production traits in a Landrace-Duroc-Yorkshire population

    Directory of Open Access Journals (Sweden)

    Lea A. Rempel

    2011-05-01

    Full Text Available The ovary and placenta are dynamic structures requiring constant modification both structurally and through cell-cell communication capabilities. The extracellular matrix and basement membranes are primarily composed of a milieu of glycosaminoglycans, including heparan sulfate and hyaluronan. Heparanase (HPSE and hyaluronidases (HYAL are responsible for degrading heparan sulfate and hyaluronan, respectively. Therefore, the objective of this study was to evaluate the relationship of SNPs distinct to HPSE, HYAL1, and HYAL2 with measurements of reproduction and production traits in swine. Single trait associations were performed on a Landrace-Duroc-Yorkshire population using SNPs discovered and identified in HPSE, HYAL1, and HYAL2. Analyses were conducted on an extended pedigree and SNPs were found to be associated with reproductive and production traits. Prior to multiple-testing corrections, SNPs within HPSE were weakly associated (P < 0.03 having additive effects with age at puberty (-2.5d + 1.08, ovulation rate (0.5 corpora lutea + 0.24, and number of piglets born alive (0.9 piglets + 0.44. A HYAL1 and two HYAL2 SNP were nominally associated (P < 0.0063 with number of piglets born alive after multiple-testing corrections (effects between 1.02 and 1.44 piglets, while one of the same HYAL2 markers maintained a modest association (P = 0.0043 having a dominant effect with number of piglets weaned (1.2 piglets + 0.41 after multiple-testing correction. Functionally, HPSE and HYAL1 and 2 have been shown to participate in events related to ovarian and placental activity. SNPs from these studies could potentially assist with understanding genetic components underlying sow lifetime productivity as measured by piglet survivability based on number born alive and number weaned, thereby contributing to a greater number of pigs/sow/year.

  9. [Biological characterization of three natural isolates of the porcine rubulavirus (Mexico)].

    Science.gov (United States)

    Borraz-Argüello, María del Tránsito; Santos-López, Gerardo; Vallejo-Ruiz, Verónica; Herrera-Camacho, Irma; Reyes-Leyva, Julio

    2008-06-01

    Biological characterization of three natural isolates of the porcine rubulavirus (Mexico). Porcine rubulavirus (PoRV) produces a neurological and reproductive syndrome in pigs called the blue-eye disease, known only from Mexico. Several isolates were grouped by the main symptoms presented during outbreaks: a) neurotropic in piglets, (b) broadly neurotropic in piglets and gonadotropic in adults, and (c) gonadotropic in adults. We studied some biological properties of three strains, which fall in one of each virus group: La Piedad Michoacán (LPM) and Producci6n Animal Cerdos 1 (PAC1) and 3 (PAC3), respectively. The analyzed viral properties are mainly related with the trans-membrane hemagglutinin-neuraminidase (HN) and fusion (F) proteins, such as cytopathic effect, hemolysis, hemagglutinating (HA) and neuraminidase (NA) activities. In the infection assays PAC1 strain presented the highest fusogenicity level; however, the most cytolytic strain was PAC3. In addition, HA and NA activities and viral genome of PAC3 strain was detected in supernatants during cell infection earlier than in the other two strains, which shows that PAC3 virions release from the host cell earlier than LPM and PAC1. Experimental determination in purified viruses shows that PAC3 presented a higher HA and NA activities; however, PAC1 shows other interesting properties, such as a high thermostability of HN and differences about substrate profile respect to LPM and PAC3. Our data suggest that NA activity is associated with the virulence of RVP.

  10. Glycoprotein 5 of porcine reproductive and respiratory syndrome virus strain SD16 inhibits viral replication and causes G2/M cell cycle arrest, but does not induce cellular apoptosis in Marc-145 cells

    Energy Technology Data Exchange (ETDEWEB)

    Mu, Yang, E-mail: muyang@nwsuaf.edu.cn [Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A& F University, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture of the People' s Republic of China, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Li, Liangliang, E-mail: lifeiyang2007@126.com [Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A& F University, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture of the People' s Republic of China, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Zhang, Beibei, E-mail: diana851218@163.com [Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A& F University, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture of the People' s Republic of China, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Huang, Baicheng, E-mail: hbch228@163.com [Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A& F University, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture of the People' s Republic of China, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Gao, Jiming, E-mail: jimingao2006@163.com [Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A& F University, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture of the People' s Republic of China, No. 22 Xinong Road, Yangling, Shaanxi 712100 (China); and others

    2015-10-15

    Cell apoptosis is common after infection with porcine reproductive and respiratory syndrome virus (PRRSV). PRRSV GP5 has been reported to induce cell apoptosis. To further understand the role of GP5 in PRRSV induced cell apoptosis, we established Marc-145 cell lines stably expressing full-length GP5, GP5{sup Δ84-96} (aa 84-96 deletion), and GP5{sup Δ97-119} (aa 97-119 deletion). Cell proliferation, cell cycle progression, cell apoptosis and virus replication in these cell lines were evaluated. Neither truncated nor full-length GP5 induced cell apoptosis in Marc-145 cells. However, GP5{sup Δ97-119}, but not full-length or GP5{sup Δ84-96}, induced a cell cycle arrest at the G2/M phase resulting in a reduction in the growth of Marc-145 cells. Additionally, GP5{sup Δ84-96} inhibited the replication of PRRSV in Marc-145 cells through induction of IFN-β. These findings suggest that PRRSV GP5 is not responsible for inducing cell apoptosis in Marc-145 cells under these experimental conditions; however it has other important roles in virus/host cell biology. - Highlights: • Marc-145 cell lines stable expression PRRSV GP5 or truncated GP5 were constructed. • GP5{sup Δ97-119} expression in Marc-145 cell induced cell cycle arrest at G2/M phase. • Expression of GP5 and truncated GP5 could not induce Marc-145 cells apoptosis. • PRRSV replication in Marc-145-GP5{sup Δ84-96} was significantly inhibited.

  11. A Review of Quantitative Tools Used to Assess the Epidemiology of Porcine Reproductive and Respiratory Syndrome in U.S. Swine Farms Using Dr. Morrison’s Swine Health Monitoring Program Data

    Directory of Open Access Journals (Sweden)

    Carles Vilalta

    2017-06-01

    Full Text Available Porcine reproductive and respiratory syndrome (PRRS causes far-reaching financial losses to infected countries and regions, including the U.S. The Dr. Morrison’s Swine Health Monitoring Program (MSHMP is a voluntary initiative in which producers and veterinarians share sow farm PRRS status weekly to contribute to the understanding, in quantitative terms, of PRRS epidemiological dynamics and, ultimately, to support its control in the U.S. Here, we offer a review of a variety of analytic tools that were applied to MSHMP data to assess disease dynamics in quantitative terms to support the decision-making process for veterinarians and producers. Use of those methods has helped the U.S. swine industry to quantify the cyclical patterns of PRRS, to describe the impact that emerging pathogens has had on that pattern, to identify the nature and extent at which environmental factors (e.g., precipitation or land cover influence PRRS risk, to identify PRRS virus emerging strains, and to assess the influence that voluntary reporting has on disease control. Results from the numerous studies reviewed here provide important insights into PRRS epidemiology that help to create the foundations for a near real-time prediction of disease risk, and, ultimately, will contribute to support the prevention and control of, arguably, one of the most devastating diseases affecting the North American swine industry. The review also demonstrates how different approaches to analyze and visualize the data may help to add value to the routine collection of surveillance data and support infectious animal disease control.

  12. Evaluation of a DNA vaccine candidate co-expressing GP3 and GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) with interferon α/γ in immediate and long-lasting protection against HP-PRRSV challenge.

    Science.gov (United States)

    Du, Yijun; Qi, Jing; Lu, Yu; Wu, Jiaqiang; Yoo, Dongwan; Liu, Xing; Zhang, Xiumei; Li, Jun; Sun, Wenbo; Cong, Xiaoyan; Shi, Jianli; Wang, Jinbao

    2012-12-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) has become one of the most economically important diseases to the global pork industry. Current vaccination strategies only provide a limited protective efficacy. In this study, a DNA vaccine, pVAX1(©)-α-γ-GP35, co-expressing GP3 and GP5 of PRRSV with interferon α/γ was constructed, and its immediate and long-lasting protection against highly pathogenic PRRSV (HP-PRRSV) challenge were examined in pigs. For immediate protection, the results showed that pVAX1(©)-α-γ-GP35 could provide partially protective efficacy, which was similar to the pVAX1(©)-α-γ (expressing interferon α/γ). For long-lasting protection, pigs inoculated with pVAX1(©)-α-γ-GP35 developed significantly higher PRRSV-specific antibody response, T cell proliferation, IFN-γ, and IL-4, than those vaccinated with pVAX1(©)-GP35 (expressing GP3 and GP5 of PRRSV). Following homologous challenge with HP-PRRSV strain SD-JN, pigs inoculated with pVAX1(©)-α-γ-GP35 showed almost no clinical signs, no lung lesions, and significantly lower viremia, as compared to those in pVAX1(©)-GP35 group. It indicated that pVAX1(©)-α-γ-GP35 could induce enhanced immune responses and provide both immediate and long-lasting protection against HP-PRRSV challenge in pigs. The DNA vaccine pVAX1(©)-α-γ-GP35 might be an attractive candidate vaccine for the prevention and control of HP-PRRSV infections.

  13. Potentiation of Taishan Pinus massoniana pollen polysaccharide on the immune response and protection elicited by a highly pathogenic porcine reproductive and respiratory syndrome virus glycoprotein 5 subunit in pigs.

    Science.gov (United States)

    Peng, Jun; Yuan, Yanmei; Du, Yijun; Wu, Jiaqiang; Li, Baoquan; Li, Jun; Yu, Jiang; Hu, Liping; Shen, Si; Wang, Jinbao; Zhu, Ruiliang

    2016-04-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) heavily affects the global pork industry. Current available vaccine strategies have inherent drawbacks. In this work, the immune enhancement from Taishan Pinus massoniana pollen polysaccharide (TPPPS) and Freund's adjuvant on the efficacy of a PRRSV subunit vaccine were examined. Titers of specific anti-highly pathogenic PRRSV (HP-PRRSV) ELISA antibody and neutralizing antibody were significantly higher in pigs from the groups inoculated with medium- and high-dose TPPPS (mTPPPS, hTPPPS) adjuvant co-administered with a recombinant HP-PRRSV glycoprotein 5 subunit (GP5) than those from other groups (P 0.05). The ratio between CD3(+)CD4(+) and CD3(+)CD8(+) T lymphocyte subpopulations indicated the inoculums of GP5 + mTPPPS and GP5 + hTPPPS induced consistently higher CD3(+)CD4(+) T lymphocyte subpopulations than other inoculums (P 0.05). The low-dose TPPPS (lTPPPS) adjuvant also exhibited enhancement effects on humoral immune and T lymphocyte proliferation responses but these were significantly lower than the mTPPPS and hTPPPS doses (P < 0.05). Pigs challenged with HP-PRRSV from the GP5 + mTPPPS, GP5 + hTPPPS, and GP5 + Freund's adjuvant groups showed lower viremia, fewer clinical signs, and fewer pathological lung lesions compared with the groups of GP5-alone and GP5 + lTPPPS (P < 0.05). There were significant differences between the GP5-alone and GP5 + lTPPPS groups in detection indexes after viral challenge (P < 0.05). In conclusion, moderate doses of TPPPS as an adjuvant with GP5 show promise as a candidate for a HP-PRRSV subunit vaccine to efficiently prevent and control HP-PRRSV. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Glycosyl-phosphatidylinositol (GPI)-anchored membrane association of the porcine reproductive and respiratory syndrome virus GP4 glycoprotein and its co-localization with CD163 in lipid rafts

    Energy Technology Data Exchange (ETDEWEB)

    Du, Yijun [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan (China); Pattnaik, Asit K. [School of Veterinary Medicine and Biomedical Sciences and the Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583-0900 (United States); Song, Cheng [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Yoo, Dongwan, E-mail: dyoo@illinois.edu [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Li, Gang, E-mail: dyoo@illinois.edu [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing (China)

    2012-03-01

    The porcine reproductive and respiratory syndrome virus (PRRSV) glycoprotein 4 (GP4) resembles a typical type I membrane protein in its structure but lacks a hydrophilic tail at the C-terminus, suggesting that GP4 may be a lipid-anchored membrane protein. Using the human decay-accelerating factor (DAF; CD55), a known glycosyl-phosphatidylinositol (GPI) lipid-anchored protein, chimeric constructs were made to substitute the GPI-anchor domain of DAF with the putative lipid-anchor domain of GP4, and their membrane association and lipase cleavage were determined in cells. The DAF-GP4 fusion protein was transported to the plasma membrane and was cleaved by phosphatidylinositol-specific phospholipase C (PI-PLC), indicating that the C-terminal domain of GP4 functions as a GPI anchor. Mutational studies for residues adjacent to the GPI modification site and characterization of respective mutant viruses generated from infectious cDNA clones show that the ability of GP4 for membrane association corresponded to virus viability and growth characteristics. The residues T158 ({omega} - 2, where {omega} is the GPI moiety at E160), P159 ({omega} - 1), and M162 ({omega} + 2) of GP4 were determined to be important for virus replication, with M162 being of particular importance for virus infectivity. The complete removal of the peptide-anchor domain in GP4 resulted in a complete loss of virus infectivity. The depletion of cholesterol from the plasma membrane of cells reduced the virus production, suggesting a role of lipid rafts in PRRSV infection. Remarkably, GP4 was found to co-localize with CD163 in the lipid rafts on the plasma membrane. Since CD163 has been reported as a cellular receptor for PRRSV and GP4 has been shown to interact with this receptor, our data implicates an important role of lipid rafts during entry of the virus.

  15. Intranasal immunization of pigs with porcine reproductive and respiratory syndrome virus-like particles plus 2', 3'-cGAMP VacciGrade™ adjuvant exacerbates viremia after virus challenge.

    Science.gov (United States)

    Van Noort, Alexandria; Nelsen, April; Pillatzki, Angela E; Diel, Diego G; Li, Feng; Nelson, Eric; Wang, Xiuqing

    2017-04-12

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes reproductive failure in pregnant sows and acute respiratory disease in young pigs. It is a leading infectious agent of swine respiratory complex, which has significant negative economic impact on the swine industry. Commercial markets currently offer both live attenuated and killed vaccines; however, increasing controversy exists about their efficacy providing complete protection. Virus-like particles (VLPs) possess many desirable features of a potent vaccine candidate and have been proven to be highly immunogenic and protective against virus infections. Here we explored the efficacy of PRRSV VLPs together with the use of a novel 2', 3'-cGAMP VacciGrade™ adjuvant. Animals were immunized twice intranasally with phosphate buffered saline (PBS), PRRSV VLPs, or PRRSV VLPs plus 2', 3'-cGAMP VacciGrade™ at 2 weeks apart. Animals were challenged with PRRSV-23983 at 2 weeks post the second immunization. PRRSV specific antibody response and cytokines were measured. Viremia, clinical signs, and histological lesions were evaluated. PRRSV N protein specific antibody was detected in all animals at day 10 after challenge, but no significant difference was observed among the vaccinated and control groups. Surprisingly, a significantly higher viremia was observed in the VLPs and VLPs plus the adjuvant groups compared to the control group. The increased viremia is correlated with a higher interferon-α induction in the serum of the VLPs and the VLPs plus the adjuvant groups. Intranasal immunizations of pigs with PRRSV VLPs and VLPs plus the 2', 3'-cGAMP VacciGrade™ adjuvant exacerbates viremia. A higher level of interferon-α production, but not interferon-γ and IL-10, is correlated with enhanced virus replication. Overall, PRRSV VLPs and PRRSV VLPs plus the adjuvant fail to provide protection against PRRSV challenge. Different dose of VLPs and alternative route of vaccination such as intramuscular

  16. Analysis of ORF5 and full-length genome sequences of porcine reproductive and respiratory syndrome virus isolates of genotypes 1 and 2 retrieved worldwide provides evidence that recombination is a common phenomenon and may produce mosaic isolates.

    Science.gov (United States)

    Martín-Valls, G E; Kvisgaard, L K; Tello, M; Darwich, L; Cortey, M; Burgara-Estrella, A J; Hernández, J; Larsen, L E; Mateu, E

    2014-03-01

    Recombination is currently recognized as a factor for high genetic diversity, but the frequency of such recombination events and the genome segments involved are not well known. In the present study, we initially focused on the detection of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) isolates by examining previously published data sets of ORF5 sequences (genotypes 1 and 2) obtained worldwide. We then examined full-length genome sequences in order to determine potential recombination breakpoints along the viral genome. For ORF5, 11 sets of genotype 1 sequences from different geographical areas, including 2 Asian, 1 American, and 7 European regions, and three sets of genotype 2, including sets from China, Mexico, and the United States, were analyzed separately. Potential recombination breakpoints were detected in 10/11 genotype 1 sets, including 9 cases in which the clustering of at least one isolate was different before and after the breakpoints. In genotype 2, potential breakpoints and different tree clustering of at least one strain before and after the breakpoint were observed in 2 out of 3 sets. The results indicated that most of the ORF5 data sets contained at least one recombinant sequence. When the full-length genome sequences were examined, both genotype 1 and 2 sets presented breakpoints (10 and 9, respectively), resulting in significantly different topologies before and after the breakpoints. Mosaic genomes were detected in genotype 1 sequences. These results may have significant implications for the understanding of the molecular epidemiology of PRRSV. PRRSV is one of the most important viruses affecting swine production worldwide, causing big economic losses and sanitary problems. One of the key questions on PRRSV arises from its genetic diversity, which is thought to have a direct impact on immunobiology, epidemiology, diagnosis, and vaccine efficacy. One of the causes of this genetic diversity is recombination among strains

  17. Cellular cholesterol is required for porcine nidovirus infection.

    Science.gov (United States)

    Jeon, Ji Hyun; Lee, Changhee

    2017-12-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine epidemic diarrhea virus (PEDV) are porcine nidoviruses that are considered emerging and re-emerging viral pathogens of pigs that pose a significant economic threat to the global pork industry. Although cholesterol is known to affect the replication of a broad range of viruses in vitro, its significance and role in porcine nidovirus infection remains to be elucidated. Therefore, the present study was conducted to determine whether cellular or/and viral cholesterol levels play a role in porcine nidovirus infection. Our results showed that depletion of cellular cholesterol by treating cells with methyl-β-cyclodextrin (MβCD) dose-dependently suppressed the replication of both nidoviruses. Conversely, cholesterol depletion from the viral envelope had no inhibitory effect on porcine nidovirus production. The addition of exogenous cholesterol to MβCD-treated cells moderately restored the infectivity of porcine nidoviruses, indicating that the presence of cholesterol in the target cell membrane is critical for viral replication. The antiviral activity of MβCD on porcine nidovirus infection was found to be predominantly exerted when used as a treatment pre-infection or prior to the viral entry process. Furthermore, pharmacological sequestration of cellular cholesterol efficiently blocked both virus attachment and internalization and, accordingly, markedly affected subsequent post-entry steps of the replication cycle, including viral RNA and protein biosynthesis and progeny virus production. Taken together, our data indicate that cell membrane cholesterol is required for porcine nidovirus entry into cells, and pharmacological drugs that hamper cholesterol-dependent virus entry may have antiviral potential against porcine nidoviruses.

  18. Virulence Attributes of Low-Virulence Organisms

    Directory of Open Access Journals (Sweden)

    Bryan Larsen

    1994-01-01

    virulence organisms present in the female lower genital tract, we are beginning to identify some of their virulence attributes. Examples from the work of our laboratory include the hemolysin of Gardnerella vaginalis and an immunosuppressive mycotoxin produced by Candida albicans. Demonstrating the coordinate expression (or other control mechanisms of virulence factors in these sometimes innocuous and sometimes inimical organisms represents the next frontier in the study of normal vaginal microbiology.

  19. Dioxin exposure and porcine reproductive hormonal activity

    Directory of Open Access Journals (Sweden)

    Gregoraszczuk Ewa L.

    2002-01-01

    Full Text Available To characterize the action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD during both the follicular and luteal phases of the ovarian cycle, the direct effect of TCDD was investigated in vitro using a system of primary monolayer cell culture. Granulosa and theca cells were collected from the preovulatory follicles and cultured as a co-culture, thus resembling follicles in vivo. Luteal cells were isolated from the corpora lutea collected during the midluteal phase. In both cases cells were isolated from the ovaries of animals exhibiting natural estrus cycle. Results of these experiments suggest that TCDD decreases estradiol secretion by follicular cells and progesterone secretion by luteal cells in a dose-dependent manner. It was also shown that TCDD disrupts steroidogenesis through its influence on the activity of enzymes involved in the steroid biosynthesis cascade. In luteal cells, its action is mediated via the aryl hydrocarbon receptor (AhR and is probably independent of estrogen receptor (ER stimulation. Endocrine disruptors that interfere with estradiol production in the follicles can act as ovulatory disruptors, and while interfering with progesterone production by luteal cells they can act as abortifacients.

  20. Brucella spp. Virulence Factors and Immunity.

    Science.gov (United States)

    Byndloss, Mariana X; Tsolis, Renee M

    2016-01-01

    Brucellosis, caused by bacteria of the genus Brucella, is an important zoonotic infection that causes reproductive disease in domestic animals and chronic debilitating disease in humans. An intriguing aspect of Brucella infection is the ability of these bacteria to evade the host immune response, leading to pathogen persistence. Conversely, in the reproductive tract of infected animals, this stealthy pathogen is able to cause an acute severe inflammatory response. In this review, we discuss the different mechanisms used by Brucella to cause disease, with emphasis on its virulence factors and the dichotomy between chronic persistence and reproductive disease.

  1. Investigation of SNPs in the porcine desmoglein 1 gene

    DEFF Research Database (Denmark)

    Daugaard, L.; Andresen, Lars Ole; Fredholm, M.

    2007-01-01

    Background: Desmoglein I (DSGI) is the target protein in the skin disease exudative epidermitis in pigs caused by virulent strains of Staphylococcus hyicus. The exfoliative toxins produced by S. hyicus digest the porcine desmoglein I (PIG)DSGI by a very specific reaction. This study investigated...... the location of single nucleotide polymorphisms (SNPs) in the porcine desmoglein I gene (PIG)DSGI in correlation to the cleavage site as well as if the genotype of the SNPs is correlated to susceptibility or resistance to the disease. Results: DNA from 32 affected and 32 unaffected piglets with exudative...... the genotypes of two out of seven SNPs found in the porcine desmoglein I gene and the susceptibility to exudative epidermitis....

  2. Blood gas and hematological changes in experimental peracute porcine pleuropneumonia.

    OpenAIRE

    Kiorpes, A L; MacWilliams, P S; Schenkman, D I; Bäckström, L R

    1990-01-01

    The effect of experimental, peracute, porcine pleuropneumonia on arterial blood gases, acid base status, the leukogram, and gross and microscopic lung structure was studied in nine growing pigs (mean weight +/- SD 10.6 +/- 2.0 kg). Pigs were inoculated intranasally with a virulent serotype 5 isolate of Actinobacillus pleuropneumoniae, and all showed signs typical of the disease within four hours. Death occurred in all pigs from 4.5 to 32 hours postinoculation (mean 14 hours). Gross and micros...

  3. Neuraminidase activity of blue eye disease porcine rubulavirus: Specificity, affinity and inhibition studies.

    Science.gov (United States)

    Santos-López, Gerardo; Borraz-Argüello, María T; Márquez-Domínguez, Luis; Flores-Alonso, Juan Carlos; Ramírez-Mendoza, Humberto; Priem, Bernard; Fort, Sébastien; Vallejo-Ruiz, Verónica; Reyes-Leyva, Julio; Herrera-Camacho, Irma

    2017-10-01

    Porcine rubulavirus (PorPV), also known as La Piedad Michoacan Virus (LPMV) causes encephalitis and reproductive failure in newborn and adult pigs, respectively. The hemagglutinin-neuraminidase (HN) glycoprotein is the most exposed and antigenic of the virus proteins. HN plays central roles in PorPV infection; i.e., it recognizes sialic acid-containing cell receptors that mediate virus attachment and penetration; in addition, its neuraminidase (sialic acid releasing) activity has been proposed as a virulence factor. This work describes the purification and characterization of PorPV HN protein (isolate PAC1). The specificity of neuraminidase is restricted to sialyl(α2,3)lactose (3SL). HN showed typical Michaelis-Menten kinetics with fetuin as substrate (km=0.029μM, Vmax=522.8nmolmin-1mg-1). When 3SL was used as substrate, typical cooperative kinetics were found (S50=0.15μM, Vmax=154.3nmolmin-1mg-1). The influenza inhibitor zanamivir inhibited the PorPV neuraminidase with IC50 of 0.24μM. PorPV neuraminidase was activated by Ca2+ and inhibited by nucleoside triphosphates with the level of inhibition depending on phosphorylation level. The present results open possibilities to study the role of neuraminidase in the pathogenicity of PorPV infection and its potential inhibitors. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. An Investigation of the Pathology and Pathogens Associated with Porcine Respiratory Disease Complex in Denmark

    DEFF Research Database (Denmark)

    Hansen, Mette Sif; Pors, S. E.; Jensen, H. E.

    2010-01-01

    Respiratory infections are among the most important diseases of growing pigs. In order to elucidate the multifactorial aetiology of porcine respiratory disease complex (PRDC) in Denmark, lungs from 148 finishing pigs with cranioventral bronchopneumonia (case group) and 60 pigs without lung lesions......), porcine reproductive and respiratory syndrome virus (both European and US type), porcine circovirus type 2 (PCV2), porcine respiratory coronavirus, porcine cytomegalovirus, Mycoplasma hyopneumoniae and Mycoplasma hyorhinis. All cases had cranioventral lobular bronchopneumonia consistent with PRDC....... There was a broad range of microscopical lesions and the cases were characterized as acute (n=10), subacute (n=24) or chronic (n=114) bronchopneumonia. Five bacterial species, five viruses and two Mycoplasma spp. were detected in different combinations. PCV2, M. hyopneumoniae, M. hyorhinis and Pasteurella multocida...

  5. Circovirose suína Porcine circovirosis: a review

    Directory of Open Access Journals (Sweden)

    Ticiana do Nascimento França

    2005-06-01

    Full Text Available Por meio de revisão da literatura pertinente foram coligidos e são apresentados os principais dados relativos aos aspectos epidemiológicos, clínicos, anátomo e histopatológicos observados na infecção por Circovírus Porcino tipo 2 (PCV-2 em suínos. São abordados a Síndrome Definhante Multissistêmica dos Suínos Desmamados (SDMDS, o Tremor Congênito Suíno (TCS, a Síndrome da Nefropatia e Dermatite Porcina (SNDP, bem como outras enfermidades associadas ou correlatas, a Síndrome Respiratória e Reprodutiva Porcina (SRRP, a Pneumonia Necrotizante Proliferativa (PNP e as falhas reprodutivas. Uma vez que a SDMSD já foi registrada na Região Sul do Brasil e no Estado do Rio de Janeiro esse estudo objetiva chamar a atenção para o especial significado dessa virose para a suinocultura brasileira, em função dos prejuízos econômicos por ela determinados.The literature of Porcine Circovirosis, including the main data on epidemiology and clinical, macroscopic and microscopic alterations of the infection of swine by Porcine Circovirus type 2 (PCV-2, is reviewed. There are various forms of infection: the [Porcine] Postweaning Multisystemic Wasting Syndrome (PMWS, Porcine Congenital Tremor, Porcine Dermatitis and Nephropathy Syndrome, and other associated or correlated diseases as the Porcine Reproductive and Respiratory Syndrome, Proliferative Necrotizing Pneumonia, and reproductive disorders. As PMWS already has been reported from southern Brazil and from the state of Rio de Janeiro, the objective of this review is to draw attention to the implications of this virosis for swine production in Brazil and its economical importance.

  6. Beyond Mortality: Sterility As a Neglected Component of Parasite Virulence.

    Science.gov (United States)

    Abbate, Jessica L; Kada, Sarah; Lion, Sébastien

    2015-12-01

    Virulence is generally defined as the reduction in host fitness following infection by a parasite (see Box 1 for glossary) [1]. In general, parasite exploitation of host resources may reduce host survival (mortality virulence), decrease host fecundity (sterility virulence), or even have sub-lethal effects that disturb the way individuals interact within a community (morbidity) [2,3]. In fact, the virulence of many parasites involves a combination of these various effects (Box 2). In practice, however, virulence is most often defined as disease-induced mortality [1, 4-6]. This is especially true in the theoretical literature, where the evolution of sterility virulence, morbidity, and mixed strategies of host exploitation have received relatively little attention. While the focus on mortality effects has allowed for easy comparison between models and, thus, rapid advancement of the field, we ask whether these theoretical simplifications have led us to inadvertently minimize the evolutionary importance of host sterilization and secondary virulence effects. As explicit theoretical work on morbidity is currently lacking (but see [7]), our aim in this Opinion piece is to discuss what is understood about sterility virulence evolution, its adaptive potential, and the implications for parasites that utilize a combination of host survival and reproductive resources.

  7. Blood gas and hematological changes in experimental peracute porcine pleuropneumonia.

    Science.gov (United States)

    Kiorpes, A L; MacWilliams, P S; Schenkman, D I; Bäckström, L R

    1990-01-01

    The effect of experimental, peracute, porcine pleuropneumonia on arterial blood gases, acid base status, the leukogram, and gross and microscopic lung structure was studied in nine growing pigs (mean weight +/- SD 10.6 +/- 2.0 kg). Pigs were inoculated intranasally with a virulent serotype 5 isolate of Actinobacillus pleuropneumoniae, and all showed signs typical of the disease within four hours. Death occurred in all pigs from 4.5 to 32 hours postinoculation (mean 14 hours). Gross and microscopic changes were typical of porcine pleuropneumonia in all pigs. Changes in the leukogram included a rapid decline in total white cells, segmented neutrophils, lymphocytes, monocytes, and eosinophils. Pigs maintained alveolar ventilation throughout the study as arterial CO2 tension was unchanged; however, arterial O2 tension and pH decreased from (mean +/- SD) 95.2 +/- 5.7 torr and 7.463 +/- 0.018 at baseline to 62.1 +/- 12.3 torr and 7.388 +/- 0.045, respectively, within 90 minutes prior to death. The data showed that in this model of peracute porcine pleuropneumonia, progressive ventilatory failure was not a feature of the disease, and the blood gas values and acid base status were maintained within physiological ranges. The histopathological hematological and physiological findings were consistent with the hypothesis that peracute porcine pleuropneumonia resembles septic shock. Images Fig. 2. Fig. 3. PMID:2106382

  8. Porcine embryonic stem cells

    DEFF Research Database (Denmark)

    Hall, Vanessa Jane

    2008-01-01

    The development of porcine embryonic stem cell lines (pESC) has received renewed interest given the advances being made in the production of immunocompatible transgenic pigs. However, difficulties are evident in the production of pESCs in-vitro. This may largely be attributable to differences...

  9. Porcine SLITRK1

    DEFF Research Database (Denmark)

    Larsen, Knud Erik; Momeni, Jamal; Farajzadeh, Leila

    2014-01-01

    The membrane protein SLITRK1 functions as a developmentally regulated stimulator of neurite outgrowth and variants in this gene have been implicated in Tourette syndrome. In the current study we have cloned and characterized the porcine SLITRK1 gene. The genomic organization of SLITRK1 lacks intr...

  10. Effects of human and porcine bile on the proteome of Helicobacter hepaticus

    Directory of Open Access Journals (Sweden)

    Okoli Arinze S

    2012-04-01

    Full Text Available Abstract Background Helicobacter hepaticus colonizes the intestine and liver of mice causing hepatobiliary disorders such as hepatitis and hepatocellular carcinoma, and has also been associated with inflammatory bowel disease in children. In its habitat, H. hepaticus must encounter bile which has potent antibacterial properties. To elucidate virulence and host-specific adaptation mechanisms of H. hepaticus modulated by human or porcine bile, a proteomic study of its response to the two types of bile was performed employing two-dimensional gel electrophoresis (2-DE and mass spectrometry. Results The 2-DE and mass spectrometry analyses of the proteome revealed that 46 proteins of H. hepaticus were differentially expressed in human bile, 18 up-regulated and 28 down-regulated. In the case of porcine bile, 32 proteins were differentially expressed of which 19 were up-regulated, and 13 were down-regulated. Functional classifications revealed that identified proteins participated in various biological functions including stress response, energy metabolism, membrane stability, motility, virulence and colonization. Selected genes were analyzed by RT-PCR to provide internal validation for the proteomic data as well as provide insight into specific expressions of motility, colonization and virulence genes of H. hepaticus in response to human or porcine bile. Conclusions Overall, the data suggested that bile is an important factor that determines virulence, host adaptation, localization and colonization of specific niches within host environment.

  11. Bacterial proteases and virulence

    DEFF Research Database (Denmark)

    Frees, Dorte; Brøndsted, Lone; Ingmer, Hanne

    2013-01-01

    Bacterial pathogens rely on proteolysis for variety of purposes during the infection process. In the cytosol, the main proteolytic players are the conserved Clp and Lon proteases that directly contribute to virulence through the timely degradation of virulence regulators and indirectly by providing....... These extracellular proteases are activated in complex cascades involving auto-processing and proteolytic maturation. Thus, proteolysis has been adopted by bacterial pathogens at multiple levels to ensure the success of the pathogen in contact with the human host....

  12. Isolation and open reading frame 5 gene analysis of porcine ...

    African Journals Online (AJOL)

    ARL

    2012-11-08

    Nov 8, 2012 ... Isolation and open reading frame 5 gene analysis of porcine reproductive and respiratory syndrome virus in. Yunnan Province, China. WANG Qing-lu1, ZHANG Shu-guang1, LI Fu-xiang2, BI Bao-liang1, CHAI Jun1,. WANG Sheng-kui1* and ZHANG Yi-fang1*. 1College of Animal Science and Technology, ...

  13. Clonal analysis and virulent traits of pathogenic extraintestinal Escherichia coli isolates from swine in China

    Directory of Open Access Journals (Sweden)

    Ding Yi

    2012-08-01

    Full Text Available Abstract Background Extraintestinal pathogenic Escherichia coli (ExPEC can cause a variety of infections outside the gastrointestinal tract in humans and animals. Infections due to swine ExPECs have been occurring with increasing frequency in China. These ExPECs may now be considered a new food-borne pathogen that causes cross-infections between humans and pigs. Knowledge of the clonal structure and virulence genes is needed as a framework to improve the understanding of phylogenetic traits of porcine ExPECs. Results Multilocus sequence typing (MLST data showed that the isolates investigated in this study could be placed into four main clonal complexes, designated as CC10, CC1687, CC88 and CC58. Strains within CC10 were classified as phylogroup A, and these accounted for most of our porcine ExPEC isolates. Isolates in the CC1687 clonal complex, formed by new sequence types (STs, was classified as phylogroup D, with CC88 isolates considered as B2 and CC58 isolates as B1. Porcine ExPECs in these four clonal complexes demonstrated significantly different virulence gene patterns. A few porcine ExPECs were indentified in phylogroup B2, the phylogroup in which human ExPECs mainly exist. However some STs in the four clonal groups of porcine ExPECs were reported to cause extraintestinal infections in human, based on data in the MLST database. Conclusion Porcine ExPECs have different virulence gene patterns for different clonal complexes. However, these strains are mostly fell in phylogenentic phylogroup A, B1 and D, which is different from human ExPECs that concentrate in phylogroup B2. Our findings provide a better understanding relating to the clonal structure of ExPECs in diseased pigs and indicate a need to re-evaluate their contribution to human ExPEC diseases.

  14. Use of an inactivated vaccine for prevention of parvovirus-induced reproductive failure in gilts.

    Science.gov (United States)

    Brown, T T; Whitacre, M D; Robison, O W

    1987-01-15

    Gilts from dams that had been inoculated with inactivated porcine parvovirus (PPV) vaccine before breeding became seronegative to PPV by 26 weeks of age. Vaccination of these gilts with inactivated PPV vaccine at 32 weeks of age resulted in an antibody response that peaked at about 2 weeks after vaccination, with -log10 mean hemagglutination inhibiting (HI) antibody titers of less than 2. In the first-year group (82 gilts), HI titers gradually decreased, 20% of the gilts being seronegative by 6 to 7 weeks after vaccination and 75% being seronegative by 16 weeks after vaccination. In the second-year group, 93 gilts were infected naturally by a field strain of PPV at about 11 weeks after single vaccination with inactivated PPV. Additionally, in the second year, 20 vaccinated and 6 nonvaccinated gilts were immune-challenged with virulent PPV at 10 to 12 weeks after vaccination. Neither field nor challenge PPV infection of vaccinated pregnant gilts caused reproductive failure, even though some of the gilts became seronegative for PPV before challenge. Our findings suggest that single vaccination of gilts with inactivated PPV vaccine should give adequate protection from PPV-induced reproductive failure, even though serum HI titers decrease to an undetectable level shortly before PPV infection.

  15. Novel porcine repetitive elements

    Directory of Open Access Journals (Sweden)

    Nonneman Dan J

    2006-12-01

    Full Text Available Abstract Background Repetitive elements comprise ~45% of mammalian genomes and are increasingly known to impact genomic function by contributing to the genomic architecture, by direct regulation of gene expression and by affecting genomic size, diversity and evolution. The ubiquity and increasingly understood importance of repetitive elements contribute to the need to identify and annotate them. We set out to identify previously uncharacterized repetitive DNA in the porcine genome. Once found, we characterized the prevalence of these repeats in other mammals. Results We discovered 27 repetitive elements in 220 BACs covering 1% of the porcine genome (Comparative Vertebrate Sequencing Initiative; CVSI. These repeats varied in length from 55 to 1059 nucleotides. To estimate copy numbers, we went to an independent source of data, the BAC-end sequences (Wellcome Trust Sanger Institute, covering approximately 15% of the porcine genome. Copy numbers in BAC-ends were less than one hundred for 6 repeat elements, between 100 and 1000 for 16 and between 1,000 and 10,000 for 5. Several of the repeat elements were found in the bovine genome and we have identified two with orthologous sites, indicating that these elements were present in their common ancestor. None of the repeat elements were found in primate, rodent or dog genomes. We were unable to identify any of the replication machinery common to active transposable elements in these newly identified repeats. Conclusion The presence of both orthologous and non-orthologous sites indicates that some sites existed prior to speciation and some were generated later. The identification of low to moderate copy number repetitive DNA that is specific to artiodactyls will be critical in the assembly of livestock genomes and studies of comparative genomics.

  16. Investigation of SNPs in the porcine desmoglein 1 gene

    Directory of Open Access Journals (Sweden)

    Andresen Lars

    2007-03-01

    Full Text Available Abstract Background Desmoglein 1 (DSG1 is the target protein in the skin disease exudative epidermitis in pigs caused by virulent strains of Staphylococcus hyicus. The exfoliative toxins produced by S. hyicus digest the porcine desmoglein 1 (PIGDSG1 by a very specific reaction. This study investigated the location of single nucleotide polymorphisms (SNPs in the porcine desmoglein 1 gene (PIGDSG1 in correlation to the cleavage site as well as if the genotype of the SNPs is correlated to susceptibility or resistance to the disease. Results DNA from 32 affected and 32 unaffected piglets with exudative epidermitis were diagnosed clinically as affected or unaffected. Two regions of the desmoglein 1 gene were sequenced and genotypes of the SNPs were established. Seven SNPs (823T>C, 828A>G, 829A>G, 830A>T, 831A>T, 838A>C and 1139C>T were found in the analysed sequences and the allele frequencies were determined for the SNPs resulting in amino acid change. Four of the seven polymorphisms were situated in the motif known to be important for toxin cleavage. The distribution of the genotypes between affected and unaffected animals was analysed. Conclusion The study indicated a possible correlation between the genotypes of two out of seven SNPs found in the porcine desmoglein 1 gene and the susceptibility to exudative epidermitis.

  17. Serial transvaginal ultrasound-guided biopsy of the porcine corpus luteum in vivo

    NARCIS (Netherlands)

    Björkman, S.; Yun, J.; Niku, M.; Oliviero, C.; Soede, N.M.; Peltoniemi, O.A.T.

    2017-01-01

    The aims of the present study was to develop and describe a transvaginal ultrasound-guided biopsy method for luteal tissue in the porcine and to evaluate the effects of the method on the reproductive tract, ovarian status and pregnancy status. Biopsies were performed in four multiparous sows on Days

  18. Porcine prion protein amyloid.

    Science.gov (United States)

    Hammarström, Per; Nyström, Sofie

    2015-01-01

    Mammalian prions are composed of misfolded aggregated prion protein (PrP) with amyloid-like features. Prions are zoonotic disease agents that infect a wide variety of mammalian species including humans. Mammals and by-products thereof which are frequently encountered in daily life are most important for human health. It is established that bovine prions (BSE) can infect humans while there is no such evidence for any other prion susceptible species in the human food chain (sheep, goat, elk, deer) and largely prion resistant species (pig) or susceptible and resistant pets (cat and dogs, respectively). PrPs from these species have been characterized using biochemistry, biophysics and neurobiology. Recently we studied PrPs from several mammals in vitro and found evidence for generic amyloidogenicity as well as cross-seeding fibril formation activity of all PrPs on the human PrP sequence regardless if the original species was resistant or susceptible to prion disease. Porcine PrP amyloidogenicity was among the studied. Experimentally inoculated pigs as well as transgenic mouse lines overexpressing porcine PrP have, in the past, been used to investigate the possibility of prion transmission in pigs. The pig is a species with extraordinarily wide use within human daily life with over a billion pigs harvested for human consumption each year. Here we discuss the possibility that the largely prion disease resistant pig can be a clinically silent carrier of replicating prions.

  19. Towards the establishment of a porcine model to study human amebiasis.

    Directory of Open Access Journals (Sweden)

    Fabienne Girard-Misguich

    Full Text Available BACKGROUND: Entamoeba histolytica is an important parasite of the human intestine. Its life cycle is monoxenous with two stages: (i the trophozoite, growing in the intestine and (ii the cyst corresponding to the dissemination stage. The trophozoite in the intestine can live as a commensal leading to asymptomatic infection or as a tissue invasive form producing mucosal ulcers and liver abscesses. There is no animal model mimicking the whole disease cycle. Most of the biological information on E. histolytica has been obtained from trophozoite adapted to axenic culture. The reproduction of intestinal amebiasis in an animal model is difficult while for liver amebiasis there are well-described rodent models. During this study, we worked on the assessment of pigs as a new potential model to study amebiasis. METHODOLOGY/PRINCIPAL FINDINGS: We first co-cultured trophozoites of E. histolytica with porcine colonic fragments and observed a disruption of the mucosal architecture. Then, we showed that outbred pigs can be used to reproduce some lesions associated with human amebiasis. A detailed analysis was performed using a washed closed-jejunal loops model. In loops inoculated with virulent amebas a severe acute ulcerative jejunitis was observed with large hemorrhagic lesions 14 days post-inoculation associated with the presence of the trophozoites in the depth of the mucosa in two out four animals. Furthermore, typical large sized hepatic abscesses were observed in the liver of one animal 7 days post-injection in the portal vein and the liver parenchyma. CONCLUSIONS: The pig model could help with simultaneously studying intestinal and extraintestinal lesion development.

  20. Transient virulence of emerging pathogens.

    Science.gov (United States)

    Bolker, Benjamin M; Nanda, Arjun; Shah, Dharmini

    2010-05-06

    Should emerging pathogens be unusually virulent? If so, why? Existing theories of virulence evolution based on a tradeoff between high transmission rates and long infectious periods imply that epidemic growth conditions will select for higher virulence, possibly leading to a transient peak in virulence near the beginning of an epidemic. This transient selection could lead to high virulence in emerging pathogens. Using a simple model of the epidemiological and evolutionary dynamics of emerging pathogens, along with rough estimates of parameters for pathogens such as severe acute respiratory syndrome, West Nile virus and myxomatosis, we estimated the potential magnitude and timing of such transient virulence peaks. Pathogens that are moderately evolvable, highly transmissible, and highly virulent at equilibrium could briefly double their virulence during an epidemic; thus, epidemic-phase selection could contribute significantly to the virulence of emerging pathogens. In order to further assess the potential significance of this mechanism, we bring together data from the literature for the shapes of tradeoff curves for several pathogens (myxomatosis, HIV, and a parasite of Daphnia) and the level of genetic variation for virulence for one (myxomatosis). We discuss the need for better data on tradeoff curves and genetic variance in order to evaluate the plausibility of various scenarios of virulence evolution.

  1. Langerhans cells in porcine skin.

    Science.gov (United States)

    Nfon, Charles K; Dawson, Harry; Toka, Felix N; Golde, William T

    2008-12-15

    Langerhans cells (LCs) are resident dendritic cells (DCs) of skin and mucosal epithelium. The standard for identifying skin DCs as LCs is expression of langerin (CD207), a surface protein that mediates Birbeck granule (BG) formation upon internalization. Reports of BGs in porcine skin DC are contradictory, due to lack of langerin detection. Here, we present the sequence of porcine langerin/CD207, showing that the predicted porcine protein shares 75%/86% amino acid identity/similarity with human. Langerin mRNA was detected in porcine skin DCs by PCR and langerin protein was detected in both isolated skin DCs and skin sections by immunostaining. Approximately, 50-70% of skin DCs expressed langerin, demonstrating that the majority of porcine skin DCs are LCs. The full length sequence combined with the identification of antibodies reactive with porcine langerin, facilitates the study of LCs in swine, and advances the use of swine for studying skin diseases and infectious disease processes involving skin.

  2. Biology of Porcine Parvovirus (Ungulate parvovirus 1

    Directory of Open Access Journals (Sweden)

    István Mészáros

    2017-12-01

    Full Text Available Porcine parvovirus (PPV is among the most important infectious agents causing infertility in pigs. Until recently, it was thought that the virus had low genetic variance, and that prevention of its harmful effect on pig fertility could be well-controlled by vaccination. However, at the beginning of the third millennium, field observations raised concerns about the effectiveness of the available vaccines against newly emerging strains. Subsequent investigations radically changed our view on the evolution and immunology of PPV, revealing that the virus is much more diverse than it was earlier anticipated, and that some of the “new” highly virulent isolates cannot be neutralized effectively by antisera raised against “old” PPV vaccine strains. These findings revitalized PPV research that led to significant advancements in the understanding of early and late viral processes during PPV infection. Our review summarizes the recent results of PPV research and aims to give a comprehensive update on the present understanding of PPV biology.

  3. Virulence and Pre-Lethal Reproductive Effects of Metarhizium anisopliae var. anisopliae on Pseudococcus viburni (Hemiptera: Pseudococcidae Virulencia y Efectos Pre-Letales en la Reproducción de Metarhizium anisopliae var. anisopliae en Pseudococcus viburni (Hemiptera: Pseudococcidae

    Directory of Open Access Journals (Sweden)

    Aldo Pereira

    2011-12-01

    Full Text Available Obscure mealybug, Pseudococcus viburni (Signoret, is a cosmopolitan pest that causes damage by suction of vascular juices and the production of honeydew, as well as for being a quarantine insect. Within control options, entomopathogenic fungi are a good alternative, nevertheless, more research is needed. In this research, the Metarhizium anisopliae var. anisopliae (Metschnikoff isolate Qu-M984 was evaluated on P. viburni under laboratory conditions. Virulence was evaluated by lethal dose 50 (LD50 and lethal time 50 (LT50, for each of the four life stages of the female. The doses tested were 10(5, 10(6, 10(7 and 10(8 conidia mL-1. Also fecundity, egg size, fertility, and longevity of adult females were evaluated at doses of 10(5 and 10(6 conidia mL-1, the evaluations were made every 2 d throughout the insect life time. The LD50 and LT50 obtained were variable for each life stage, although without statistical differences among life stages (P > 0.05, ranging from LD50 = 7.3 x 10(5 to 4.9 x 10(9 conidia mL-1 and LT50 = 7.74 to 9.97 d at 10(8 conidia mL-1. Significant differences (P 0.05.Chanchito blanco de la vid, Pseudococcus viburni (Signoret, es una plaga cosmopolita que causa daños tanto por succión de jugos vasculares como por su producción de mielecilla, así como también por ser un insecto cuarentenario. Dentro de las opciones de control, hongos entomopatógenos son una buena alternativa, sin embargo, más investigación es necesaria. En esta investigación fue evaluado Metarhizium anisopliae var. anisopliae (Metschnikoff aislamiento Qu-M984 en P. viburni bajo condiciones de laboratorio. Fue evaluada virulencia según dosis letal 50 (LD50 y tiempo letal 50 (LT50 para cada uno de los cuatro estados de desarrollo de la hembra. Las dosis evaluadas fueron 10(5, 10(6, 10(7 y 10(8 conidias mL-1. Fecundidad, tamaño de huevos, fertilidad y longevidad de hembras adultas fueron evaluados con dosis 10(5 y 10(6 conidias mL-1, las evaluaciones fueron

  4. Efficacy of the porcine species in biomedical research

    Directory of Open Access Journals (Sweden)

    Karina eGutierrez

    2015-09-01

    Full Text Available Since domestication, pigs have been used extensively in agriculture and kept as companion animals. More recently they have been used in biomedical research, given they share many physiological and anatomical similarities with humans. Recent technological advances in assisted reproduction, somatic cell cloning, stem cell culture, genome editing and transgenesis now enable the creation of unique porcine models of human diseases. Here we highlight the potential applications and advantages of using pigs, particularly minipigs, as indispensable large animal models in fundamental and clinical research, including the development of therapeutics for inherited and chronic disorders, and cancers.

  5. 7 CFR 1230.611 - Porcine animal.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Porcine animal. 1230.611 Section 1230.611 Agriculture... CONSUMER INFORMATION Procedures for the Conduct of Referendum Definitions § 1230.611 Porcine animal. The term Porcine animal means a swine, that is raised: (a) As a feeder pig, that is, a young pig sold to...

  6. The impact of resource quality on the evolution of virulence in spatially heterogeneous environments.

    Science.gov (United States)

    Su, Min; Boots, Mike

    2017-03-07

    Understanding the drivers of parasite evolution and in particular disease virulence remains a major focus of evolutionary theory. Here, we examine the role of resource quality and in particular spatial environmental heterogeneity in the distribution of these resources on the evolution of virulence. There may be direct effects of resources on host susceptibility and pathogenicity alongside effects on reproduction that indirectly impact host-parasite population dynamics. Therefore, we assume that high resource quality may lead to both increased host reproduction and/or increased disease resistance. In completely mixed populations there is no effect of resource quality on the outcome of disease evolution. However, when there are local interactions higher resource quality generally selects for higher virulence/transmission for both linear and saturating transmission-virulence trade-off assumptions. The exception is that in castrators (i.e., infected hosts have no reproduction), higher virulence is selected for both low and high resource qualities at mixed local and global infection. Heterogeneity in the distribution of environment resources only has an effect on the outcome in castrators where random distributions generally select for higher virulence. Overall, our results further underline the importance of considering spatial structure in order to understand evolutionary processes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Brucella, nitrogen and virulence.

    Science.gov (United States)

    Ronneau, Severin; Moussa, Simon; Barbier, Thibault; Conde-Álvarez, Raquel; Zuniga-Ripa, Amaia; Moriyon, Ignacio; Letesson, Jean-Jacques

    2016-08-01

    The brucellae are α-Proteobacteria causing brucellosis, an important zoonosis. Although multiplying in endoplasmic reticulum-derived vacuoles, they cause no cell death, suggesting subtle but efficient use of host resources. Brucellae are amino-acid prototrophs able to grow with ammonium or use glutamate as the sole carbon-nitrogen source in vitro. They contain more than twice amino acid/peptide/polyamine uptake genes than the amino-acid auxotroph Legionella pneumophila, which multiplies in a similar vacuole, suggesting a different nutritional strategy. During these two last decades, many mutants of key actors in nitrogen metabolism (transporters, enzymes, regulators, etc.) have been described to be essential for full virulence of brucellae. Here, we review the genomic and experimental data on Brucella nitrogen metabolism and its connection with virulence. An analysis of various aspects of this metabolism (transport, assimilation, biosynthesis, catabolism, respiration and regulation) has highlighted differences and similarities in nitrogen metabolism with other α-Proteobacteria. Together, these data suggest that, during their intracellular life cycle, the brucellae use various nitrogen sources for biosynthesis, catabolism and respiration following a strategy that requires prototrophy and a tight regulation of nitrogen use.

  8. Tachykinins in the porcine pancreas

    DEFF Research Database (Denmark)

    Schmidt, P T; Tornøe, K; Poulsen, Steen Seier

    2000-01-01

    The localization, release, and effects of substance P and neurokinin A were studied in the porcine pancreas and the localization of substance P immunoreactive nerve fibers was examined by immunohistochemistry. The effects of electrical vagus stimulation and capsaicin infusion on tachykinin release...... and the effects of substance P and neurokinin A infusion on insulin, glucagon, somatostatin, and exocrine secretion were studied using the isolated perfused porcine pancreas with intact vagal innervation. NK-1 and NK-2 receptor antagonists were used to investigate receptor involvement. Substance P immunoreactive...

  9. Reproductive emergencies.

    Science.gov (United States)

    Jutkowitz, L Ari

    2005-03-01

    The emergency clinician is frequently called on to manage problems relating to the female reproductive tract. Because owners sel-dom have the medical knowledge needed to differentiate normal from abnormal reproductive behaviors, they frequently look to the emergency veterinarian for guidance and information during and after parturition. For this reason, it is essential that the veterinarian have a good understanding of the normal reproductive cycle as well as the common emergencies that may occur. This article reviews the events surrounding normal parturition in the dog and cat and the reproductive emergencies seen most commonly in practice.

  10. First report of Porcine teschovirus (PTV), Porcine sapelovirus (PSV) and Enterovirus G (EV-G) in pig herds of Brazil.

    Science.gov (United States)

    Donin, Daiane Güllich; de Arruda Leme, Raquel; Alfieri, Alice Fernandes; Alberton, Geraldo Camilo; Alfieri, Amauri Alcindo

    2014-03-01

    Porcine teschovirus (PTV), Porcine sapelovirus (PSV) and Enterovirus G (EV-G) have been associated with enteric, respiratory, reproductive and neurological disorders. Although Brazil is the world's fourth largest producer and exporter of pork, no information on the occurrence of PTV, PSV and EV-G infections is available for Brazilian pig herds. This study aimed to investigate the occurrence of Porcine enteric picornavirus infections in pig farms located in three distinct geographical regions of Brazil. Forty randomly selected diarrhoeic and normal consistency faeces of suckling (n = 22) and nursery (n = 18) pigs from farms located in 21 distinct cities of the Southern, Southeast, and Midwest regions of Brazil were evaluated by nested-RT-PCR assays. Suckling piglets presented the expected amplicon size for PTV (158 bp) and EV-G (313 bp) in single and mixed infections in 40.9 % (9/22) of the faecal samples. PSV amplicon (212 bp) was not detected in this age group. For nursery pigs, Porcine enteric picornaviruses amplicons were present in 77.8 % (14/18) of the faecal samples. PTV and EV-G were detected in single and mixed infections, while PSV was detected only in two samples in co-infection with PTV and EV-G in this age group. The Brazilian regions evaluated presented at least two of the tested viruses. Sequencing analysis revealed high similarities to the related viruses (95.3 to 99.2 % for PTV, 94.2 to 98.5 % for PSV and 86 to 100 % for EV-G). For the first time PTV, PSV and EV-G have been molecularly detected and characterised in pig faecal samples in Brazil.

  11. Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2

    Directory of Open Access Journals (Sweden)

    Yang Yang

    2017-01-01

    Full Text Available Porcine circovirus virus type II (PCV2 is the etiology of postweaning multisystemic wasting syndrome (PMWS, porcine dermatitis, nephropathy syndrome (PDNS, and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA assays using a real-time fluorescent detection (PCV2 real-time RPA assay and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 102 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 102 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2.

  12. Dimethyl Sulfoxide Perturbs Cell Cycle Progression and Spindle Organization in Porcine Meiotic Oocytes.

    Directory of Open Access Journals (Sweden)

    Xuan Li

    Full Text Available Meiotic maturation of mammalian oocytes is a precisely orchestrated and complex process. Dimethyl sulfoxide (DMSO, a widely used solvent, drug, and cryoprotectant, is capable of disturbing asymmetric cytokinesis of oocyte meiosis in mice. However, in pigs, DMSO's effect on oocyte meiosis still remains unknown. We aimed to evaluate if DMSO treatment will affect porcine oocyte meiosis and the underlying molecular changes as well. Interestingly, we did not observe the formation of the large first polar body and symmetric division for porcine oocytes treated with DMSO, contrary to findings reported in mice. 3% DMSO treatment could inhibit cumulus expansion, increase nuclear abnormality, disturb spindle organization, decrease reactive oxygen species level, and elevate mitochondrial membrane potential of porcine oocytes. There was no effect on germinal vesicle breakdown rate regardless of DMSO concentration. 3% DMSO treatment did not affect expression of genes involved in spindle organization (Bub1 and Mad2 and apoptosis (NF-κB, Pten, Bcl2, Caspase3 and Caspase9, however, it significantly decreased expression levels of pluripotency genes (Oct4, Sox2 and Lin28 in mature oocytes. Therefore, we demonstrated that disturbed cumulus expansion, chromosome alignment, spindle organization and pluripotency gene expression could be responsible for DMSO-induced porcine oocyte meiotic arrest and the lower capacity of subsequent embryo development. Our results provide new insights on DMSO's effect on porcine oocyte meiosis and raise safety concerns over DMSO's usage on female reproduction in both farm animals and humans.

  13. Swine torque teno virus detection in pig commercial vaccines, enzymes for laboratory use and human drugs containing components of porcine origin.

    Science.gov (United States)

    Kekarainen, Tuija; Martínez-Guinó, Laura; Segalés, Joaquim

    2009-03-01

    Torque teno viruses (TTVs) are vertebrate infecting, single-stranded circular DNA viruses. Two genetically distinct TTV genogroups (TTV1 and TTV2) infect swine worldwide with high prevalence. Currently, swine TTVs are considered non-pathogenic, although TTV2 has been linked to post-weaning multisystemic wasting syndrome, a porcine circovirus disease. On the other hand, pig materials are an important source of components used in porcine vaccine manufacturing, human drugs and commercial enzyme products. However, there is little information about the possible existence of extraneous viruses in products containing porcine-derived components. In the present study, 26 commercial swine vaccines, seven human drugs and three enzyme products from porcine origin were tested for the presence of TTV1 and TTV2 genomes by PCR. Four vaccines against Mycoplasma hyopneumoniae were positive for TTV2 by PCR. Three M. hyopneumoniae, one porcine parvovirus and one porcine reproductive and respiratory syndrome virus vaccines were PCR positive for TTV1. One human drug contained TTV1 DNA as well as a trypsin enzyme; a porcine-derived elastase product was positive for both TTV genogroups. These results show that swine TTVs are contaminants not only of swine vaccines but also of human drugs containing porcine components and enzymes for laboratory use.

  14. Reproductive Health and Reproductive Vulnerability

    Directory of Open Access Journals (Sweden)

    Bojan Žikić

    2016-03-01

    Full Text Available Reproductive health represents, almost to an equal extent, a socio-cultural and a medical fact. What influences it, both positively and negatively, stems from the ways in which we culturally cognize and act with regard to reproductive behavior. These thoughts and actions are conditioned by a culturally contextualized conceptualization of human physiology which is, in turn, based on the conceptualization of sexuality, and especially, the normativization of gender roles. Therefore, reproductive health is, above all, female health, when viewed as a socio-cultural category, meaning that reproductive vulnerability mostly refers to those factors that negatively influence female reproductive health. These factors are social – they negatively influence reproductive health through the institutional and legally normative aspects, they are economic – they decrease the number of those who, in a certain socio-cultural context, have timely access to quality medical care, and they are cultural – they reinforce modes of thinking and behavior which do not take into consideration the right of every human being to his or her own sexual and reproductive life, but rather insist on conforming individual sexuality and reproductive desires and capacities to the dominant cultural norm.

  15. Rapid evolution of virulence leading to host extinction under host-parasite coevolution.

    Science.gov (United States)

    Rafaluk, Charlotte; Gildenhard, Markus; Mitschke, Andreas; Telschow, Arndt; Schulenburg, Hinrich; Joop, Gerrit

    2015-06-13

    Host-parasite coevolution is predicted to result in changes in the virulence of the parasite in order to maximise its reproductive success and transmission potential, either via direct host-to-host transfer or through the environment. The majority of coevolution experiments, however, do not allow for environmental transmission or persistence of long lived parasite stages, in spite of the fact that these may be critical for the evolutionary success of spore forming parasites under natural conditions. We carried out a coevolution experiment using the red flour beetle, Tribolium castaneum, and its natural microsporidian parasite, Paranosema whitei. Beetles and their environment, inclusive of spores released into it, were transferred from generation to generation. We additionally took a modelling approach to further assess the importance of transmissive parasite stages on virulence evolution. In all parasite treatments of the experiment, coevolution resulted in extinction of the host population, with a pronounced increase in virulence being seen. Our modelling approach highlighted the presence of environmental transmissive parasite stages as being critical to the trajectory of virulence evolution in this system. The extinction of host populations was unexpected, particularly as parasite virulence is often seen to decrease in host-parasite coevolution. This, in combination with the increase in virulence and results obtained from the model, suggest that the inclusion of transmissive parasite stages is important to improving our understanding of virulence evolution.

  16. Meloidogyne virulence locus molecular marker for characterization of selected mi-virulent populations of Meloidogyne spp. is correlated with several genera of betaproteobacteria.

    Science.gov (United States)

    Cortada, Laura; Sakai, Hiromichi; Verdejo-Lucas, Soledad; Mizukubo, Takayuki

    2011-04-01

    Resistance to root-knot nematodes in tomato is conferred by the Mi resistance gene to the three most important species of Meloidogyne: M. arenaria, M. incognita, and M. javanica. Nevertheless, the Mi gene is unable to inhibit the reproduction of selected and naturally Mi-virulent populations of root-knot nematodes. As pathogenicity assays are time consuming, molecular markers were developed for the easy identification of Mi-virulent populations of Meloidogyne. The sequence characterized amplified region-Meloidogyne virulence locus (MVC) molecular marker is reported to differentiate Mi-avirulent and naturally Mi-virulent from selected Mi-virulent populations. This marker was used to compare acquired virulence in populations of M. javanica from Spain. The original populations used to develop the MVC marker were included as control for reference. Results showed that this marker did not amplify genomic DNA extracted from single juveniles or females of any of the populations tested either from Spain or Japan. In silico analyses performed with the recently published complete genome of M. incognita, indicated that the MVC marker is not correlated to a MVC or to any eukaryotic organism but to several betaproteobacteria genus from the family Comamonadaceae.

  17. Responses of tomato genotypes to avirulent and Mi-virulent Meloidogyne javanica isolates occurring in Israel.

    Science.gov (United States)

    Iberkleid, Ionit; Ozalvo, Rachel; Feldman, Lidia; Elbaz, Moshe; Patricia, Bucki; Horowitz, Sigal Brown

    2014-05-01

    The behavior of naturally virulent Meloidogyne isolates toward the tomato resistance gene Mi in major tomato-growing areas in Israel was studied for the first time. Virulence of seven selected isolates was confirmed over three successive generations on resistant (Mi-carrying) and susceptible (non-Mi-carrying) tomato cultivars. Diagnostic markers verified the predominance of Meloidogyne javanica among virulent isolates selected on resistant tomato cultivars or rootstocks. To better understand the determinants of nematode selection on Mi-carrying plants, reproduction of Mi-avirulent and virulent isolates Mjav1 and Mjv2, respectively, measured as eggs per gram of root, on non-Mi-carrying, heterozygous (Mi/mi) and homozygous (Mi/Mi) genotypes was evaluated. Although no reproduction of Mjav1 was observed on Mi/Mi genotypes, some reproduction was consistently observed on Mi/mi plants; reproduction of Mjv2 on the homozygous and heterozygous genotypes was similar to that on susceptible cultivars, suggesting a limited quantitative effect of the Mi gene. Histological examination of giant cells induced by Mi-virulent versus avirulent isolates confirmed the high virulence of Mjv2 on Mi/mi and Mi/Mi genotypes, allowing the formation of well-developed giant-cell systems despite the Mi gene. Analysis of the plant defense response in tomato Mi/Mi, Mi/mi, and mi/mi genotypes to both avirulent and virulent isolates was investigated by quantitative real-time polymerase chain reaction. Although the jasmonate (JA)-signaling pathway was clearly upregulated by avirulent and virulent isolates on the susceptible (not carrying Mi) and heterozygous (Mi/mi) plants, no change in signaling was observed in the homozygous (Mi/Mi) resistant line following incompatible interaction with the avirulent isolate. Thus, similar to infection promoted by the avirulent isolate on the susceptible genotype, the Mi-virulent isolate induced the JA-dependent pathway, which might promote tomato susceptibility

  18. Characterization and virulence clustering analysis of extraintestinal pathogenic Escherichia coli isolated from swine in China.

    Science.gov (United States)

    Zhu, Yinchu; Dong, Wenyang; Ma, Jiale; Yuan, Lvfeng; Hejair, Hassan M A; Pan, Zihao; Liu, Guangjin; Yao, Huochun

    2017-04-08

    Swine extraintestinal pathogenic Escherichia coli (ExPEC) is an important pathogen that leads to economic and welfare costs in the swine industry worldwide, and is occurring with increasing frequency in China. By far, various virulence factors have been recognized in ExPEC. Here, we investigated the virulence genotypes and clonal structure of collected strains to improve the knowledge of phylogenetic traits of porcine ExPECs in China. We isolated 64 Chinese porcine ExPEC strains from 2013 to 14 in China. By multiplex PCR, the distribution of isolates belonging to phylogenetic groups B1, B2, A and D was 9.4%, 10.9%, 57.8% and 21.9%, respectively. Nineteen virulence-related genes were detected by PCR assay; ompA, fimH, vat, traT and iutA were highly prevalent. Virulence-related genes were remarkably more prevalent in group B2 than in groups A, B1 and D; notably, usp, cnf1, hlyD, papA and ibeA were only found in group B2 strains. Genotyping analysis was performed and four clusters of strains (named I to IV) were identified. Cluster IV contained all isolates from group B2 and Cluster IV isolates had the strongest pathogenicity in a mouse infection model. As phylogenetic group B2 and D ExPEC isolates are generally considered virulent, multilocus sequence typing (MLST) analysis was performed for these isolates to further investigate genetic relationships. Two novel sequence types, ST5170 and ST5171, were discovered. Among the nine clonal complexes identified among our group B2 and D isolates, CC12 and CC95 have been indicated to have high zoonotic pathogenicity. The distinction between group B2 and non-B2 isolates in virulence and genotype accorded with MLST analysis. This study reveals significant genetic diversity among ExPEC isolates and helps us to better understand their pathogenesis. Importantly, our data suggest group B2 (Cluster IV) strains have the highest risk of causing animal disease and illustrate the correlation between genotype and virulence.

  19. 7 CFR 1230.18 - Porcine animal.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Porcine animal. 1230.18 Section 1230.18 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING AGREEMENTS... animal. Porcine animal means a swine, that is raised as (a) a feeder pig, that is, a young pig sold to...

  20. Differences in virulence of Naegleria fowleri.

    Science.gov (United States)

    De Jonckheere, J

    1979-10-01

    All pathogenic Naegleria fowleri isolated from the environment were highly virulent to mice when instilled intranasally. Axenic cultivation gradually decreased virulence of highly virulent strains. This decrease was most pronounced in environmental isolates and of minor importance in N. fowleri isolated from human cerebrospinal fluid. The low virulent strains obtained by continuous axenic cultivation appeared after clonation to consist of individuals with different virulence. Virulence could be enhanced in low virulent strains by brain passage and passages in Vero cell cultures, but could not be induced by these methods in nonvirulent strains isolated from the environment. Different mice strains showed different sensitivities to infection with pathogenic Naegleria. In addition, older mice were less sensitive than younger animals to low virulent strains.

  1. Complete Genome Sequence of a Porcine Epidemic Diarrhea Virus Strain from Vietnam, HUA-14PED96, with a Large Genomic Deletion.

    Science.gov (United States)

    Choe, Se-Eun; Park, Kee-Hwan; Lim, Seong-In; Le, Van Phan; Hien, Nguyen Ba; Thach, Pham Ngoc; Phuong, Le Huynh Thanh; An, Byung-Hyun; Han, Song Hee; Cho, In-Soo; An, Dong-Jun

    2016-02-18

    A highly virulent strain of Porcine epidemic diarrhea virus (PEDV) causing severe diarrhea has recently emerged in Vietnam. Genomic sequences from a novel strain, HUA-14PED96, isolated from a Vietnamese piglet with serious diarrhea show relatively high identity with U.S.-like PEDV strains, and have a 72-nt deletion in the open reading frame 1a (ORF1a) gene. Copyright © 2016 Choe et al.

  2. Antimicrobial compounds of porcine mucosa

    Science.gov (United States)

    Kotenkova, E. A.; Lukinova, E. A.; Fedulova, L. V.

    2017-09-01

    The aim of the study was to investigate porcine oral cavity mucosa (OCM), nasal cavity mucosa (NCM), rectal mucosa (RM) and tongue mucosa (TM) as sources of antimicrobial compounds. Ultrafiltrates with MW >30 kDa, MW 5-30 kDa and MW antimicrobial activity against Escherichia coli and Proteus vulgaris. NCM ultrafiltrates revealed the highest antibacterial activity in respect to negative control: for the fraction with MW >30 kDa, the zone of microbial growth inhibition was 7.5 mm, for the MWantimicrobial compounds, which could be an actual alternative for reduction of microbial spoilage of foods.

  3. Inheritance of porcine stress syndrome.

    Science.gov (United States)

    Mabry, J W; Christian, L L; Kuhlers, D L

    1981-01-01

    A total of 66 litters were farrowed in a Yorkshire herd of pigs selected for porcine stress syndrome (PSS) susceptibility. These litters included all possible combinations of matings between stress-susceptible, stress-carrier, and stress-resistant animals. When the data were analyzed by within-litter chi-square analysis, the null hypothesis of recessive inheritance could not be rejected (P less than 0.05). In addition, when the data were analyzed across litters, the null hypothesis of autosomal recessive inheritance could not be rejected (P less than 0.05).

  4. Preliminary findings of a previously unrecognized porcine primary immunodeficiency disorder.

    Science.gov (United States)

    Ozuna, A G Cino; Rowland, R R R; Nietfeld, J C; Kerrigan, M A; Dekkers, J C M; Wyatt, C R

    2013-01-01

    Weaned pigs from a line bred for increased feed efficiency were enrolled in a study of the role of host genes in the response to infection with Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). Four of the pigs were euthanatized early in the study due to weight loss with illness and poor body condition; 2 pigs before PRRSV infection and the other 2 pigs approximately 2 weeks after virus inoculation. The 2 inoculated pigs failed to produce PRRSV-specific antibodies. Gross findings included pneumonia, absence of a detectable thymus, and small secondary lymphoid tissues. Histologically, lymph nodes, spleen, tonsils, and Peyer's patches were sparsely cellular with decreased to absent T and B lymphocytes.

  5. Transcriptomic Analysis Reveals Selective Metabolic Adaptation of Streptococcus suis to Porcine Blood and Cerebrospinal Fluid

    Directory of Open Access Journals (Sweden)

    Anna Koczula

    2017-02-01

    Full Text Available Streptococcus suis is a zoonotic pathogen that can cause severe pathologies such as septicemia and meningitis in its natural porcine host as well as in humans. Establishment of disease requires not only virulence of the infecting strain but also an appropriate metabolic activity of the pathogen in its host environment. However, it is yet largely unknown how the streptococcal metabolism adapts to the different host niches encountered during infection. Our previous isotopologue profiling studies on S. suis grown in porcine blood and cerebrospinal fluid (CSF revealed conserved activities of central carbon metabolism in both body fluids. On the other hand, they suggested differences in the de novo amino acid biosynthesis. This prompted us to further dissect S. suis adaptation to porcine blood and CSF by RNA deep sequencing (RNA-seq. In blood, the majority of differentially expressed genes were associated with transport of alternative carbohydrate sources and the carbohydrate metabolism (pentose phosphate pathway, glycogen metabolism. In CSF, predominantly genes involved in the biosynthesis of branched-chain and aromatic amino acids were differentially expressed. Especially, isoleucine biosynthesis seems to be of major importance for S. suis in CSF because several related biosynthetic genes were more highly expressed. In conclusion, our data revealed niche-specific metabolic gene activity which emphasizes a selective adaptation of S. suis to host environments.

  6. Transcriptomic Analysis Reveals Selective Metabolic Adaptation of Streptococcus suis to Porcine Blood and Cerebrospinal Fluid.

    Science.gov (United States)

    Koczula, Anna; Jarek, Michael; Visscher, Christian; Valentin-Weigand, Peter; Goethe, Ralph; Willenborg, Jörg

    2017-02-15

    Streptococcus suis is a zoonotic pathogen that can cause severe pathologies such as septicemia and meningitis in its natural porcine host as well as in humans. Establishment of disease requires not only virulence of the infecting strain but also an appropriate metabolic activity of the pathogen in its host environment. However, it is yet largely unknown how the streptococcal metabolism adapts to the different host niches encountered during infection. Our previous isotopologue profiling studies on S. suis grown in porcine blood and cerebrospinal fluid (CSF) revealed conserved activities of central carbon metabolism in both body fluids. On the other hand, they suggested differences in the de novo amino acid biosynthesis. This prompted us to further dissect S. suis adaptation to porcine blood and CSF by RNA deep sequencing (RNA-seq). In blood, the majority of differentially expressed genes were associated with transport of alternative carbohydrate sources and the carbohydrate metabolism (pentose phosphate pathway, glycogen metabolism). In CSF, predominantly genes involved in the biosynthesis of branched-chain and aromatic amino acids were differentially expressed. Especially, isoleucine biosynthesis seems to be of major importance for S. suis in CSF because several related biosynthetic genes were more highly expressed. In conclusion, our data revealed niche-specific metabolic gene activity which emphasizes a selective adaptation of S. suis to host environments.

  7. Reproductive epidemiology

    DEFF Research Database (Denmark)

    Olsen, Jørn; Nøhr, Ellen Aagaard

    2010-01-01

    Reproductive health covers a broad category of health and disease conditions, according to the Cairo Statement. This chapter focuses on subfecundity fertility, fetal death, malformations, pregnancy complications, sexual health, and diseases that may have their origin in fetal life, but which...

  8. Reproductive physiology

    Science.gov (United States)

    Gee, G.F.; Russman, S.E.; Ellis, David H.; Gee, George F.; Mirande, Claire M.

    1996-01-01

    Conclusions: Although the general pattern of avian physiology applies to cranes, we have identified many physiological mechanisms (e.g., effects of disturbance) that need further study. Studies with cranes are expensive compared to those done with domestic fowl because of the crane's larger size, low reproductive rate, and delayed sexual maturity. To summarize, the crane reproductive system is composed of physiological and anatomical elements whose function is controlled by an integrated neural-endocrine system. Males generally produce semen at a younger age than when females lay eggs. Eggs are laid in clutches of two (1 to 3), and females will lay additional clutches if the preceding clutches are removed. Both sexes build nests and incubate the eggs. Molt begins during incubation and body molt may be completed annually in breeding pairs. However, remiges are replaced sequentially over 2 to 3 years, or abruptly every 2 to 3 years in other species. Most immature birds replace their juvenal remiges over a 2 to 3 year period. Stress interferes with reproduction in cranes by reducing egg production or terminating the reproductive effort. In other birds, stress elevates corticosterone levels and decreases LHRH release. We know little about the physiological response of cranes to stress.

  9. Genetic diversity, virulotyping and antimicrobial resistance susceptibility of Yersinia enterocolitica isolated from pigs and porcine products in Malaysia.

    Science.gov (United States)

    Thong, Kwai Lin; Tan, Lai Kuan; Ooi, Peck Toung

    2018-01-01

    The objectives of the present study were to determine the antimicrobial resistance, virulotypes and genetic diversity of Yersinia enterocolitica isolated from uncooked porcine food and live pigs in Malaysia. Thirty-two non-repeat Y. enterocolitica strains of three bioserotypes (3 variant/O:3, n = 27; 1B/O:8, n = 3; 1A/O:5, n = 2) were analysed. Approximately 90% of strains were multidrug-resistant with a multiple antibiotic resistance index Yersinia enterocolitica could be distinguished distinctly into three clusters by pulsed-field gel electrophoresis, with each belonging to a particular bioserotype. Strains of 3 variant/O:3 were more heterogeneous than others. Eleven of the 15 virulence genes tested (hreP, virF, rfbC, myfA, sat, inv, ail, ymoA, ystA, tccC, yadA) and pYV virulence plasmid were present in all the bioserotpe 3 variant/03 strains. The occurrence of virulent strains of Y. enterocolitica in pigs and porcine products reiterated that pigs are important reservoirs for Y. enterocolitica. The increasing trend of multidrug resistant strains is a public health concern. This is the first report on the occurrence of potential pathogenic and resistant strains of Y. enterocolitica in pigs in Malaysia. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  10. Tribological evaluation of porcine skin.

    Science.gov (United States)

    Xiao, Huaping; Ariyasinghe, Nethika; He, Xingliang; Liang, Hong

    2014-04-01

    This research studies the effects of external parameters on the friction of porcine skin. A tribometer was used to evaluate the frictional behavior of the same. The effects of DI water and body oil on porcine skin against steel and glass balls were evaluated in terms of coefficient of friction (COF). The COF dropped rapidly when DI water/body oil was introduced into the sliding system and remained stable when the volume of the liquid exceeded a certain value. The COF increased with increasing sliding speed under dry conditions and decreased in wet. Under an increasing normal force, the COF decreased regardless of the presence of liquid. The ratio of the real contact area to the nominal contact area of the skin with the steel/glass ball was found to increase with a power law as the applied force was increased. These results reveal basic tribological properties of the skin in contact with a hard slider. These properties could be used as reference for the design and development of artificial skin in prosthetic applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Existence of proviral porcine endogenous retrovirus in fresh and decellularised porcine tissues

    OpenAIRE

    Prabha S; Verghese S

    2008-01-01

    Purpose: Swine are expected to be utilized as xenograft donors for both whole organ and cellular transplantation. A major concern in using porcine organs for transplantation is the potential of transmission of porcine endogenous retrovirus (PERV). Tissue-engineered or decellularised heart valves have already been implanted in humans and have been marketed by certain companies after Food and Drug Administration (FDA) approval. The aim of this study was to examine the existence of porcine endo...

  12. Men's Reproductive Health

    Science.gov (United States)

    ... Submit Cancel Close Email Share Dialog × Print Men's Reproductive Health Reproductive health is an important component of men's overall health ... often, males have been overlooked in discussions of reproductive health, especially when reproductive issues such as contraception and ...

  13. Male Reproductive System

    Science.gov (United States)

    ... Kidney Transplant Vision Facts and Myths Male Reproductive System KidsHealth > For Parents > Male Reproductive System Print A ... son's reproductive health. continue About the Male Reproductive System Most species have two sexes: male and female. ...

  14. Phenotyping and susceptibility of established porcine cells lines to African Swine Fever Virus infection and viral production.

    Science.gov (United States)

    Sánchez, Elena G; Riera, Elena; Nogal, Marisa; Gallardo, Carmina; Fernández, Paloma; Bello-Morales, Raquel; López-Guerrero, José Antonio; Chitko-McKown, Carol G; Richt, Jürgen A; Revilla, Yolanda

    2017-09-04

    African swine fever virus (ASFV) is a highly pathogenic, double-stranded DNA virus with a marked tropism for cells of the monocyte-macrophage lineage, affecting swine species and provoking severe economic losses and health threats. In the present study, four established porcine cell lines, IPAM-WT, IPAM-CD163, C∆2+ and WSL, were compared to porcine alveolar macrophage (PAM) in terms of surface marker phenotype, susceptibility to ASFV infection and virus production. The virulent ASFV Armenia/07, E70 or the naturally attenuated NHV/P68 strains were used as viral models. Cells expressed only low levels of specific receptors linked to the monocyte/macrophage lineage, with low levels of infection overall, with the exception of WSL, which showed more efficient production of strain NHV/P68 but not of strains E70 and Armenia/07.

  15. Identification of species-specific novel transcripts in pig reproductive tissues using RNA-seq

    NARCIS (Netherlands)

    Du, Z.; Eisley, C.J.; Onteru, S.K.; Madsen, O.; Groenen, M.; Ross, J.W.; Rothschild, M.F.

    2014-01-01

    Although structural properties of the porcine reproductive system are shared by many placental mammals, some combination of these properties is unique to pigs. To explore whether genomic elements specific to pigs could potentially underlie this uniqueness, we made the first step to identify novel

  16. Porcine head response to blast.

    Science.gov (United States)

    Shridharani, Jay K; Wood, Garrett W; Panzer, Matthew B; Capehart, Bruce P; Nyein, Michelle K; Radovitzky, Raul A; Bass, Cameron R 'dale'

    2012-01-01

    Recent studies have shown an increase in the frequency of traumatic brain injuries related to blast exposure. However, the mechanisms that cause blast neurotrauma are unknown. Blast neurotrauma research using computational models has been one method to elucidate that response of the brain in blast, and to identify possible mechanical correlates of injury. However, model validation against experimental data is required to ensure that the model output is representative of in vivo biomechanical response. This study exposes porcine subjects to primary blast overpressures generated using a compressed-gas shock tube. Shock tube blasts were directed to the unprotected head of each animal while the lungs and thorax were protected using ballistic protective vests similar to those employed in theater. The test conditions ranged from 110 to 740 kPa peak incident overpressure with scaled durations from 1.3 to 6.9 ms and correspond approximately with a 50% injury risk for brain bleeding and apnea in a ferret model scaled to porcine exposure. Instrumentation was placed on the porcine head to measure bulk acceleration, pressure at the surface of the head, and pressure inside the cranial cavity. Immediately after the blast, 5 of the 20 animals tested were apneic. Three subjects recovered without intervention within 30 s and the remaining two recovered within 8 min following respiratory assistance and administration of the respiratory stimulant doxapram. Gross examination of the brain revealed no indication of bleeding. Intracranial pressures ranged from 80 to 390 kPa as a result of the blast and were notably lower than the shock tube reflected pressures of 300-2830 kPa, indicating pressure attenuation by the skull up to a factor of 8.4. Peak head accelerations were measured from 385 to 3845 G's and were well correlated with peak incident overpressure (R(2) = 0.90). One SD corridors for the surface pressure, intracranial pressure (ICP), and head acceleration are

  17. Campylobacter virulence and survival factors.

    Science.gov (United States)

    Bolton, Declan J

    2015-06-01

    Despite over 30 years of research, campylobacteriosis is the most prevalent foodborne bacterial infection in many countries including in the European Union and the United States of America. However, relatively little is known about the virulence factors in Campylobacter or how an apparently fragile organism can survive in the food chain, often with enhanced pathogenicity. This review collates information on the virulence and survival determinants including motility, chemotaxis, adhesion, invasion, multidrug resistance, bile resistance and stress response factors. It discusses their function in transition through the food processing environment and human infection. In doing so it provides a fundamental understanding of Campylobacter, critical for improved diagnosis, surveillance and control. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Appraisal of the porcine kidney autotransplantation model

    NARCIS (Netherlands)

    Post, Ivo C. J. H.; Dirkes, Marcel C.; Heger, Michal; van Loon, Johannes P. A. M.; Swildens, Bas; Huijzer, Goos M.; van Gulik, Thomas M.

    2012-01-01

    Animal models are extensively used for transplantation related research, especially kidney transplantation. Porcine autotransplantation models are considered to be favorable regarding translatability to the human setting. The key determinants for translatability of the model are discussed,

  19. Unisexual reproduction of Cryptococcus gattii.

    Directory of Open Access Journals (Sweden)

    Sujal S Phadke

    Full Text Available Cryptococcus gattii is a basidiomycetous human fungal pathogen that typically causes infection in tropical and subtropical regions and is responsible for an ongoing outbreak in immunocompetent individuals on Vancouver Island and in the Pacific Northwest of the US. Pathogenesis of this species may be linked to its sexual cycle that generates infectious propagules called basidiospores. A marked predominance of only one mating type (α in clinical and environmental isolates suggests that a-α opposite-sex reproduction may be infrequent or geographically restricted, raising the possibility of an alternative unisexual cycle involving cells of only α mating type, as discovered previously in the related pathogenic species Cryptococcus neoformans. Here we report observation of hallmark features of unisexual reproduction in a clinical isolate of C. gattii (isolate 97/433 and describe genetic and environmental factors conducive to this sexual cycle. Our results are consistent with population genetic evidence of recombination in the largely unisexual populations of C. gattii and provide a useful genetic model for understanding how novel modes of sexual reproduction may contribute to evolution and virulence in this species.

  20. Appraisal of the porcine kidney autotransplantation model.

    Science.gov (United States)

    Post, Ivo C J H; Dirkes, Marcel C; Heger, Michal; van Loon, Johannes P A M; Swildens, Bas; Huijzer, Goos M; van Gulik, Thomas M

    2012-01-01

    Animal models are extensively used for transplantation related research, especially kidney transplantation. Porcine autotransplantation models are considered to be favorable regarding translatability to the human setting. The key determinants for translatability of the model are discussed, comprising animal age, development, anatomy, anesthesia and surgical protocols, and perioperative care. With the detailed discussion of these determinants and the pitfalls in diagnosing animal discomfort, an attempt is made to provide a uniform porcine kidney autotransplantation model with tools to improve currently used models.

  1. Evaluation of Galleria mellonella larvae for studying the virulence of Streptococcus suis.

    Science.gov (United States)

    Velikova, Nadya; Kavanagh, Kevin; Wells, Jerry M

    2016-12-15

    Streptococcus suis is an encapsulated Gram-positive bacterium and the leading cause of sepsis and meningitis in young pigs, resulting in considerable economic losses in the porcine industry. S. suis is considered an emerging zoonotic agent with increasing numbers of human cases over the last years. In the environment, both avirulent and virulent strains occur in pigs, with no evidence for consistent adapatation of virulent strains to the human host. Currently, there is an urgent need for a convenient, reliable and standardised animal model to rapidly assess S. suis virulence. Wax moth (Galleria mellonella) larvae have successfully been used in human and animal infectious disease studies. Here, we developed G. mellonella larvae as a model to assess virulence of S. suis strains. Fourteen isolates of S. suis belonging to different serotypes killed G. mellonella larvae in a dose-dependent manner. Larvae infected with the virulent serotype 2 strain, S. suis S3881/S10, were rescued by antibiotic therapy. Crucially, the observed virulence of the different serotypes and mutants was in agreement with virulence observed in piglets (Sus scrofa) and the zebrafish larval infection model. Infection with heat-inactivated bacteria or bacteria-free culture supernatants showed that in most cases live bacteria are needed to cause mortality in G. mellonella. The G. mellonella model is simple, cost-efficient, and raises less ethical issues than experiments on vertebrates and reduces infrastructure requirements. Furthermore, it allows experiments to be performed at the host temperature (37 °C). The results reported here, indicate that the G. mellonella model may aid our understanding of veterinary microbial pathogens such as the emerging zoonotic pathogen S. suis and generate hypotheses for testing in the target animal host. Ultimately, this might lead to the timely introduction of new effective remedies for infectious diseases. Last but not least, use of the G. mellonella

  2. Porcine head response to blast

    Directory of Open Access Journals (Sweden)

    Jay eShridharani

    2012-05-01

    Full Text Available Recent studies have shown an increase in the frequency of traumatic brain injuries related to blast exposure. However, the mechanisms that cause blast neurotrauma are unknown. Blast neurotrauma research using computational models has been one method to elucidate that response of the brain in blast, and to identify possible mechanical correlates of injury. However, model validation against experimental data is required to ensure that the model output is representative of in vivo biomechanical response. This study exposed porcine subjects to primary blast overpressures generated using a compressed-gas shock tube. Shock tube blasts were directed to the unprotected head of each animal while the lungs and thorax were protected using ballistic protective vests similar to those employed in theater. The test conditions ranged from 110-740 kPa peak incident overpressure with scaled durations from 1.3-6.9 ms and correspond approximately with a 50% injury risk for brain bleeding and apnea in a ferret model scaled to porcine exposure. The bulk head acceleration and the pressure at the surface of the head and in the cranial cavity were measured. Immediately after the blast, 5 of the 20 animals tested were apneic. Three subjects recovered without intervention within thirty seconds and the remaining two recovered within 8 minutes following bagging and administration of the respiratory stimulant doxapram. Gross examination of the brain revealed no indication of bleeding. Intracranial pressures ranged from 80-685 kPa as a result of the blast and were notably lower than the shock tube reflected pressures of 300-2830 kPa, indicating pressure attenuation by the skull up to a factor of 8.4. Peak head accelerations were measured from 385-3845 G’s and were well correlated with peak incident overpressure (R2=0.90. One standard deviation corridors for the surface pressure, intracranial pressure, and head acceleration are presented to provide experimental data for

  3. Porcine heart interatrial septum anatomy.

    Science.gov (United States)

    Holda, Mateusz K; Holda, Jakub; Koziej, Mateusz; Piatek, Katarzyna; Klimek-Piotrowska, Wieslawa

    2018-02-16

    The left-sided atrial septal pouch (SP), a recently re-discovered anatomical structure within the human interatrial septum, has emerged as a possible source of thrombi formation and a trigger for atrial fibrillation, thereby potentially increasing the risk for ischemic stroke. In many studies, the swine interatrial septum has been used as model of the human heart. Also, possible new strategies and devices for management of the SPs may first be tested in this pig model. Therefore, in this study, we aimed to evaluate swine interatrial septum morphology and to compare it with the human analog, especially in the light of SP occurrence. A total of 75 swine (Sus scrofa f. domestica) hearts were examined. The interatrial septum morphology was assessed, and SPs were measured. The most common variant of the interatrial septum was smooth septum (26.6%) followed by the patent foramen ovale channel and right SP (both 22.7%). No left or double SPs were observed. In 28.0% of all cases the fold of tissue (left septal ridge) was observed on the left side of the interatrial septum in the location where the left-sided SP should be expected. The mean length of the patent foramen ovale channel was 7.1±1.5mm. The mean right SP depth was 6.3±2.2mm, and its ostium width and height were 5.8±1.2 and 5.3±1.6mm, respectively. There are significant differences between human and porcine interatrial septum morphology that should be taken into account during experimental studies. The absence of the left SP in swine results in the inability to use porcine heart as an experimental model for left-sided SP management. Copyright © 2018 Elsevier GmbH. All rights reserved.

  4. Effect of bovine apo-lactoferrin on the growth and virulence of Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Luna-Castro, Sarahí; Aguilar-Romero, Francisco; Samaniego-Barrón, Luisa; Godínez-Vargas, Delfino; de la Garza, Mireya

    2014-10-01

    Actinobacillus pleuropneumoniae (App) is a Gram-negative bacterium that causes porcine pleuropneumonia, leading to economic losses in the swine industry. Due to bacterial resistance to antibiotics, new treatments for this disease are currently being sought. Lactoferrin (Lf) is an innate immune system glycoprotein of mammals that is microbiostatic and microbicidal and affects several bacterial virulence factors. The aim of this study was to investigate whether bovine iron-free Lf (BapoLf) has an effect on the growth and virulence of App. Two serotype 1 strains (reference strain S4074 and the isolate BC52) and a serotype 7 reference strain (WF83) were analyzed. First, the ability of App to grow in iron-charged BLf was discarded because in vivo, BapoLf sequesters iron and could be a potential source of this element favoring the infection. The minimum inhibitory concentration of BapoLf was 14.62, 11.78 and 10.56 µM for the strain BC52, S4074 and WF83, respectively. A subinhibitory concentration (0.8 µM) was tested by assessing App adhesion to porcine buccal epithelial cells, biofilm production, and the secretion and function of toxins and proteases. Decrease in adhesion (24-42 %) was found in the serotype 1 strains. Biofilm production decreased (27 %) for only the strain 4074 of serotype 1. Interestingly, biofilm was decreased (60-70 %) in the three strains by BholoLf. Hemolysis of erythrocytes and toxicity towards HeLa cells were not affected by BapoLf. In contrast, proteolytic activity in all strains was suppressed in the presence of BapoLf. Finally, oxytetracycline produced synergistic effect with BapoLf against App. Our results suggest that BapoLf affects the growth and several of the virulence factors in App.

  5. Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi

    Energy Technology Data Exchange (ETDEWEB)

    Whittingham, Jean L.; Blagova, Elena V. [University of York, Heslington, York YO10 5DD (United Kingdom); Finn, Ciaran E.; Luo, Haixia; Miranda-CasoLuengo, Raúl [University College Dublin, Dublin (Ireland); Turkenburg, Johan P.; Leech, Andrew P.; Walton, Paul H. [University of York, Heslington, York YO10 5DD (United Kingdom); Murzin, Alexey G. [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Meijer, Wim G. [University College Dublin, Dublin (Ireland); Wilkinson, Anthony J., E-mail: tony.wilkinson@york.ac.uk [University of York, Heslington, York YO10 5DD (United Kingdom)

    2014-08-01

    VapD is one of a set of highly homologous virulence-associated proteins from the multi-host pathogen Rhodococcus equi. The crystal structure reveals an eight-stranded β-barrel with a novel fold and a glycine rich ‘bald’ surface. Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-d-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins.

  6. Activity of Zearalenone in the Porcine Intestinal Tract

    Directory of Open Access Journals (Sweden)

    Magdalena Gajęcka

    2016-12-01

    Full Text Available This study demonstrates that low doses (somewhat above the No Observed Adverse Effect Level, NOAEL of the mycoestrogen zearalenone (ZEN and its metabolites display multispecificity towards various biological targets in gilts. The observed responses in gilts were surprising. The presence of ZEN and zearalenols (ZELs did not evoke a response in the porcine gastrointestinal tract, which was attributed to dietary tolerance. Lymphocyte proliferation was intensified in jejunal mesenteric lymph nodes, and lymphocyte counts increased in the jejunal epithelium with time of exposure. In the distal digestive tract, fecal bacterial counts decreased, the activity of fecal bacterial enzymes and lactic acid bacteria increased, and cecal water was characterized by higher genotoxicity. The accompanying hyperestrogenism led to changes in mRNA activity of selected enzymes (cytochrome P450, hydroxysteroid dehydrogenases, nitric oxide synthases and receptors (estrogen and progesterone receptors, and it stimulated post-translational modifications which play an important role in non-genomic mechanisms of signal transmission. Hyperestrogenism influences the regulation of the host’s steroid hormones (estron, estradiol and progesteron, it affects the virulence of bacterial genes encoding bacterial hydroxysteroid dehydrogenases (HSDs, and it participates in detoxification processes by slowing down intestinal activity, provoking energy deficits and promoting antiporter activity at the level of enterocytes. In most cases, hyperestrogenism fulfils all of the above roles. The results of this study indicate that low doses of ZEN alleviate inflammatory processes in the digestive system, in particular in the proximal and distal intestinal tract, and increase body weight gains in gilts.

  7. Differential interactions of virulent and non-virulent H. parasuis strains with naïve or swine influenza virus pre-infected dendritic cells

    Directory of Open Access Journals (Sweden)

    Mussá Tufária

    2012-11-01

    Full Text Available Abstract Pigs possess a microbiota in the upper respiratory tract that includes Haemophilus parasuis. Pigs are also considered the reservoir of influenza viruses and infection with this virus commonly results in increased impact of bacterial infections, including those by H. parasuis. However, the mechanisms involved in host innate responses towards H. parasuis and their implications in a co-infection with influenza virus are unknown. Therefore, the ability of a non-virulent H. parasuis serovar 3 (SW114 and a virulent serovar 5 (Nagasaki strains to interact with porcine bone marrow dendritic cells (poBMDC and their modulation in a co-infection with swine influenza virus (SwIV H3N2 was examined. At 1 hour post infection (hpi, SW114 interaction with poBMDC was higher than that of Nagasaki, while at 8 hpi both strains showed similar levels of interaction. The co-infection with H3N2 SwIV and either SW114 or Nagasaki induced higher levels of IL-1β, TNF-α, IL-6, IL-12 and IL-10 compared to mock or H3N2 SwIV infection alone. Moreover, IL-12 and IFN-α secretion differentially increased in cells co-infected with H3N2 SwIV and Nagasaki. These results pave the way for understanding the differences in the interaction of non-virulent and virulent strains of H. parasuis with the swine immune system and their modulation in a viral co-infection.

  8. Field trials of an inactivated virus vaccine against porcine parvovirus.

    Science.gov (United States)

    Castro, J M; del Pozo, M; Simarro, I

    1992-07-01

    Serological response and reproductive performance were estimated in field trials of an inactivated virus vaccine against porcine parvovirus. Experiments were carried out in 10 selected pig breeding herds. A total of 277 seronegative gilts were used. Two hundred and twenty animals were vaccinated twice before mating, fourteen days apart and revaccinated after farrowing. Blood samples were obtained from both vaccinated and non-vaccinated (57 animal) control gilts, one week after the 2nd dose of vaccination, at farrowing time and one week after revaccination. Although there were considerable variations among the herds, the number of returns to oestrus in all herds was higher in vaccinated gilts (11.81%) than in the controls (10.52%). This difference, however, was not statistically significant. The reproductive performance results revealed the absence of an increase in the total born, as pooled values, in vaccinated gilts compared to controls. However, when these results are interpreted in relation to serological data, many control gilts were already seropositive before mating, or remained seronegative at farrowing. According to our results, the duration of immunity with this vaccine is apparently short, as there is a clear decrease in the titres between the 1st and the 2nd sampling times (2.35 +/- 0.14 and 1.97 +/- 0.08, respectively).

  9. Selective Reproduction

    DEFF Research Database (Denmark)

    Svendsen, Mette N.

    2015-01-01

    This article employs a multi-species perspective in investigating how life's worth is negotiated in the field of neonatology in Denmark. It does so by comparing decision-making processes about human infants in the Danish neonatal intensive care unit with those associated with piglets who serve as...... as expectations within linear or predictive time frames are key markers in both sites. Exploring selective reproductive processes across human infants and research piglets can help us uncover aspects of the cultural production of viability that we would not otherwise see or acknowledge....

  10. The emergence of a new strain of porcine circovirus-2 in Ontario and Quebec swine and its association with severe porcine circovirus associated disease--2004-2006.

    Science.gov (United States)

    Carman, Susy; Cai, Hugh Y; DeLay, Josepha; Youssef, Sameh A; McEwen, Beverly J; Gagnon, Carl A; Tremblay, Donald; Hazlett, Murray; Lusis, Peter; Fairles, Jim; Alexander, Hazel S; van Dreumel, Tony

    2008-04-01

    In the late fall of 2004 more severe lesions of porcine circovirus-2 associated disease (PCVAD) than usual occurred during an outbreak of porcine circovirus-2 (PCV-2) infection in Ontario nursery and grower/finisher pigs. The lesions were of unprecedented severity and included diffuse bronchointerstitial pneumonia, granulomatous enteritis, vasculitis, interstitial nephritis, and new lesions of splenic infarction. Some affected herds had up to 50% mortality. The outbreak correlated with the sudden emergence of a variant PCV-2, with PCR restriction fragment length polymorphism (RFLP) type 321. Phylogenetic comparison of ORF2 sequences and full genome sequences showed the new variant to be different from the previously dominant RFLP type 422 viruses, and similar to viruses that had occurred in France and other European and Asian countries. A subsequent retrospective study showed a statistically significant increase in the frequency of histological lesions in lymph node, spleen, lung, small intestine, colon and kidney, for pigs spontaneously infected with RFLP type 321, compared with the older RFLP type 422 strain. Viral burden, based on IHC staining in lymph node, also showed a statistically significant increase in pigs infected with the newer variant RFLP type 321, compared with the older RFLP type 422 strain. This enhanced virulence in pigs infected with PCV-2 RFLP type 321 strain may be related to the genetic differences in this new strain of PCV-2. This virus is now the dominant strain of PCV-2 virus found in Ontario and Quebec swine.

  11. The emergence of a new strain of porcine circovirus-2 in Ontario and Quebec swine and its association with severe porcine circovirus associated disease — 2004–2006

    Science.gov (United States)

    Carman, Susy; Cai, Hugh Y.; DeLay, Josepha; A.Youssef, Sameh; McEwen, Beverly J.; Gagnon, Carl A.; Tremblay, Donald; Hazlett, Murray; Lusis, Peter; Fairles, Jim; Alexander, Hazel S.; van Dreumel, Tony

    2008-01-01

    In the late fall of 2004 more severe lesions of porcine circovirus-2 associated disease (PCVAD) than usual occurred during an outbreak of porcine circovirus-2 (PCV-2) infection in Ontario nursery and grower/finisher pigs. The lesions were of unprecedented severity and included diffuse bronchointerstitial pneumonia, granulomatous enteritis, vasculitis, interstitial nephritis, and new lesions of splenic infarction. Some affected herds had up to 50% mortality. The outbreak correlated with the sudden emergence of a variant PCV-2, with PCR restriction fragment length polymorphism (RFLP) type 321. Phylogenetic comparison of ORF2 sequences and full genome sequences showed the new variant to be different from the previously dominant RFLP type 422 viruses, and similar to viruses that had occurred in France and other European and Asian countries. A subsequent retrospective study showed a statistically significant increase in the frequency of histological lesions in lymph node, spleen, lung, small intestine, colon and kidney, for pigs spontaneously infected with RFLP type 321, compared with the older RFLP type 422 strain. Viral burden, based on IHC staining in lymph node, also showed a statistically significant increase in pigs infected with the newer variant RFLP type 321, compared with the older RFLP type 422 strain. This enhanced virulence in pigs infected with PCV-2 RFLP type 321 strain may be related to the genetic differences in this new strain of PCV-2. This virus is now the dominant strain of PCV-2 virus found in Ontario and Quebec swine. PMID:18505190

  12. Influences of ORF1 on the virulence and immunogenicity of Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Yuan, Fangyan; Liu, Jinlin; Guo, Yi; Tan, Chen; Fu, Shulin; Zhao, Jin; Chen, Huanchun; Bei, Weicheng

    2011-12-01

    Actinobacillus pleuropneumoniae is a Gram-negative pathogen that causes porcine pleuropneumonia. The pathogenicity of A. pleuropneumoniae is strongly correlated with the production of active repeat-in-toxin (RTX) proteins such as ApxIVA. We evaluated the contribution of a potential ApxIVA activator, ORF1, to the virulence and immunogenicity of A. pleuropneumoniae in pigs. The orf1 gene in A. pleuropneumoniae SLW03 (serovar 1, ΔapxICΔapxIIC) was deleted, producing strain SLW05 (ΔapxICΔapxIICΔorf1). The virulence of strains SLW03 and SLW05 was compared in pigs. Clinical signs and pulmonary lesions induced by strain SLW05 were slighter than that of strain SLW03 (P pigs immunized with strain SLW03 or SLW05 developed high antibody titers against ApxIA, ApxIIA, and ApxIVA before challenge. Two weeks after a second immunization, pigs were challenged intratracheally with either a fully virulent A. pleuropneumoniae serovar 1 or serovar 3 strain. Vaccination with strains SLW03 or SLW05 provided significantly greater protection compared to the negative control (P Immunized pigs displayed significantly fewer clinical signs and lower lung lesion scores than non-immunized pigs. These results suggested that ORF1 plays an important role in the development of ApxIVA toxicity. Furthermore, strain SLW05 is a highly attenuated strain able to induce protective immunity against A. pleuropneumoniae infection.

  13. Lichtheimia species exhibit differences in virulence potential.

    Directory of Open Access Journals (Sweden)

    Volker U Schwartze

    Full Text Available Although the number of mucormycosis cases has increased during the last decades, little is known about the pathogenic potential of most mucoralean fungi. Lichtheimia species represent the second and third most common cause of mucormycosis in Europe and worldwide, respectively. To date only three of the five species of the genus have been found to be involved in mucormycosis, namely L. corymbifera, L. ramosa and L. ornata. However, it is not clear whether the clinical situation reflects differences in virulence between the species of Lichtheimia or whether other factors are responsible. In this study the virulence of 46 strains of all five species of Lichtheimia was investigated in chicken embryos. Additionally, strains of the closest-related genus Dichotomocladium were tested. Full virulence was restricted to the clinically relevant species while all strains of L. hyalospora, L. sphaerocystis and Dichotomocladium species were attenuated. Although virulence differences were present in the clinically relevant species, no connection between origin (environmental vs clinical or phylogenetic position within the species was observed. Physiological studies revealed no clear connection of stress resistance and carbon source utilization with the virulence of the strains. Slower growth at 37°C might explain low virulence of L. hyalospora, L. spaherocystis and Dichotomocladium; however, similarly slow growing strains of L. ornata were fully virulent. Thus, additional factors or a complex interplay of factors determines the virulence of strains. Our data suggest that the clinical situation in fact reflects different virulence potentials in the Lichtheimiaceae.

  14. Structural Genomics of Bacterial Virulence Factors

    National Research Council Canada - National Science Library

    Liddington, Robert

    2004-01-01

    We are applying a comprehensive yet focused structural genomics approach to determine the atomic resolution crystal structures of key bacterial virulence factors from high priority bacterial pathogens...

  15. Virulence assessment of Portuguese isolates of potato cyst nematodes (Globodera spp.

    Directory of Open Access Journals (Sweden)

    Maria José M. DA CUNHA

    2012-05-01

    Full Text Available Identification of species and virulence groups of potato cyst nematodes (PCN, Globodera pallida and G. rostochiensis, present in field populations is important in the control of these nematodes by means of resistant cultivars. In order to characterize the virulence of Globodera spp. isolates from Portugal, 43 G. rostochiensis and three G. pallida isolates were evaluated by measuring their multiplication rates on a susceptible potato cultivar and five differential potato genotypes in a growth chamber pot experiment. Principal Component Analysis and Hierarchical Cluster Analysis showed that the reproduction rates were different in terms of both the numbers of eggs and the numbers of cysts produced. Portuguese isolates of PCN were more virulent on genotypes derived from Solanum vernei than on genotypes derived from other Solanum resistance sources, and there was a significant nematode isolate × host genotype interaction. The virulence bioassay clearly distinguished the two PCN species but failed to differentiate isolates into pathotypes. There was a wide and continuous range of virulence to the resistant genotypes, especially in G. rostochiensis isolates.

  16. Comparison of pathogenicities and nucleotide changes between porcine and bovine reassortant rotavirus strains possessing the same genotype constellation in piglets and calves.

    Science.gov (United States)

    Park, Jun-Gyu; Kim, Deok-Song; Matthijnssens, Jelle; Kwon, Hyoung-Jun; Zeller, Mark; Alfajaro, Mia Madel; Son, Kyu-Yeol; Hosmillo, Myra; Ryu, Eun-Hye; Kim, Ji-Yun; Lee, Ju-Hwan; Park, Su-Jin; Kang, Mun-Il; Kwon, Joseph; Choi, Jong-Soon; Cho, Kyoung-Oh

    2014-08-06

    Although reassortment is one of the most important characteristics of group A rotavirus (RVA) evolution, the host range restriction and/or virulence of reassortant RVAs remain largely unknown. The porcine 174-1 strain isolated from a diarrheic piglet was identified as a reassortant strain, harboring the same genotype constellation as the previously characterized bovine strain KJ56-1. Owing to its same genotype constellation, the pathogenicity of the porcine strain 174-1 in piglets and calves was examined for comparison with that of the bovine reassortant KJ56-1 strain, whose pathogenicity has already been demonstrated in piglets and calves. The porcine 174-1 strain induced diarrhea and histopathological changes in the small intestine of piglets and calves, whereas KJ56-1 had been reported to be virulent only in piglets, but not in calves. Therefore, full genomic sequences of 174-1 and KJ56-1 strains were analyzed to determine whether specific mutations might be associated with clinical and pathological phenotypes. Sequence alignment between the 174-1 and KJ56-1 strains detected one nucleotide substitution at the 3' untranslated region of the NSP3 gene and 16 amino acid substitutions at the VP7, VP4, VP1, VP3, NSP1 and NSP4 genes. These mutations may be critical molecular determinants for different virulence and/or pathogenicity of each strain. This study presents new insights into the host range restriction and/or virulence of RVAs. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Snatch-farrowed, porcine-colostrum-deprived (SF-pCD) pigs as a model for swine infectious disease research.

    Science.gov (United States)

    Huang, Yanyun; Haines, Deborah M; Harding, John C S

    2013-04-01

    The current study tested the benefit of commercially available spray-dried bovine colostrum (The Saskatoon Colostrum Company, Saskatoon, Saskatchewan) in raising snatch-farrowed, porcine-colostrum-deprived (SF-pCD) pigs. In experiment 1, 12 SF-pCD pigs received a liquid diet composed mainly of bovine colostrum from birth to day 10; 6 remained on the same liquid diet (COL), and the other 6 were fed a diet composed mainly of milk replacer (RPL) until weaning. In experiment 2, 12 SF-pCD pigs were fed mainly bovine colostrum before weaning; after weaning, 6 were fed a starter diet containing 20% (w/w) bovine colostrum powder (STARTER-COL), and the other 6 were fed a starter diet without any bovine colostrum (STARTER-CTRL) until termination (day 42 or day 49). In experiment 1 the COL pigs had significantly fewer fever-days than did the RPL pigs. In experiment 2 diarrhea, typhlocolitis, and pancreatic degeneration developed in 4 of the STARTER-COL pigs after weaning. In both experiments all the pigs fed mainly bovine colostrum before weaning survived until termination. All pigs tested free of swine influenza virus H1N1 and H3N2, Porcine reproductive and respiratory syndrome virus, and Porcine parvovirus. In experiment 2 all the pigs tested free of Porcine circovirus type 2 (PCV2), but some in both groups tested positive for Torque teno virus genogroups 1 and 2. In conclusion, with the use of snatch-farrowing and bovine colostrum, pigs can be raised in the absence of porcine maternal antibodies with 100% survival and freedom from most porcine pathogens of biologic relevance. This model is potentially suitable for animal disease research.

  18. Existence of proviral porcine endogenous retrovirus in fresh and decellularised porcine tissues

    Directory of Open Access Journals (Sweden)

    Prabha S

    2008-01-01

    Full Text Available Purpose: Swine are expected to be utilized as xenograft donors for both whole organ and cellular transplantation. A major concern in using porcine organs for transplantation is the potential of transmission of porcine endogenous retrovirus (PERV. Tissue-engineered or decellularised heart valves have already been implanted in humans and have been marketed by certain companies after Food and Drug Administration (FDA approval. The aim of this study was to examine the existence of porcine endogenous retrovirus (PERV in fresh and decellularised porcine tissues. Methods: Porcine tissues (both fresh and decellularised were analysed using validated assays specific for PERV: polymerase chain reaction (PCR, reverse transcriptase polymerase chain reaction (RT-PCR. Results: PERV specific GAG sequences were found in the porcine heart tissue samples using PCR for DNA and RT- PCR for RNA. All tissue samples (both fresh and treated tissues like aortic valve, pulmonary valve and heart muscle showed the presence of PERV DNA. RT PCR for PERV was positive in all fresh tissues and was found to be negative in decellularised treated tissues. Conclusions: PCR is a rapid, specific test for the detection of PERV virus in xenografts. These findings have demonstrated that the presence of proviral DNA form of PERV in porcine tissues needs to be carefully considered when the infectious disease potential of xenotransplantation is being assessed.

  19. Assisted Reproductive Technology (ART)

    Science.gov (United States)

    ... com/wp-content/uploads/2016/04/third-party-reproduction-booklet.pdf (PDF - 902 KB) [top] American Society for Reproductive Medicine. (2015). Assisted reproductive technologies: A guide for patients . Retrieved May ...

  20. Female Reproductive System

    Science.gov (United States)

    ... Kidney Transplant Vision Facts and Myths Female Reproductive System KidsHealth > For Parents > Female Reproductive System Print A ... or sperm. continue Parts of the Female Reproductive System Unlike the male, the human female has a ...

  1. Female reproductive disorders

    DEFF Research Database (Denmark)

    Crain, D Andrew; Janssen, Sarah J; Edwards, Thea M

    2008-01-01

    To evaluate the possible role of endocrine-disrupting compounds (EDCs) on female reproductive disorders emphasizing developmental plasticity and the complexity of endocrine-dependent ontogeny of reproductive organs. Declining conception rates and the high incidence of female reproductive...

  2. Reproduction, physiology and biochemistry

    Science.gov (United States)

    This chapter summarizes fundamental knowledge and recent discoveries about the reproduction, physiology and biochemistry of plant-parasitic nematodes. Various types of reproduction are reviewed, including sexual reproduction and mitotic and meiotic parthenogenesis. Although much is known about the p...

  3. Assisted Reproductive Technology (ART)

    Science.gov (United States)

    ... Cancel Close Email Share Dialog × Print Assisted Reproductive Technology (ART) ART refers to treatments and procedures that ... American Society for Reproductive Medicine. (2015). Assisted reproductive technologies: A guide for patients . Retrieved May 31, 2016, ...

  4. Normal Female Reproductive Anatomy

    Science.gov (United States)

    ... historical Searches are case-insensitive Reproductive System, Female, Anatomy Add to My Pictures View /Download : Small: 720x756 ... Large: 3000x3150 View Download Title: Reproductive System, Female, Anatomy Description: Anatomy of the female reproductive system; drawing ...

  5. Porcine model of hemophilia A.

    Science.gov (United States)

    Kashiwakura, Yuji; Mimuro, Jun; Onishi, Akira; Iwamoto, Masaki; Madoiwa, Seiji; Fuchimoto, Daiichiro; Suzuki, Shunichi; Suzuki, Misae; Sembon, Shoichiro; Ishiwata, Akira; Yasumoto, Atsushi; Sakata, Asuka; Ohmori, Tsukasa; Hashimoto, Michiko; Yazaki, Satoko; Sakata, Yoichi

    2012-01-01

    Hemophilia A is a common X chromosome-linked genetic bleeding disorder caused by abnormalities in the coagulation factor VIII gene (F8). Hemophilia A patients suffer from a bleeding diathesis, such as life-threatening bleeding in the brain and harmful bleeding in joints and muscles. Because it could potentially be cured by gene therapy, subhuman animal models have been sought. Current mouse hemophilia A models generated by gene targeting of the F8 have difficulties to extrapolate human disease due to differences in the coagulation and immune systems between mice and humans. Here, we generated a porcine model of hemophilia A by nuclear transfer cloning from F8-targeted fibroblasts. The hemophilia A pigs showed a severe bleeding tendency upon birth, similar to human severe hemophiliacs, but in contrast to hemophilia A mice which rarely bleed under standard breed conditions. Infusion of human factor VIII was effective in stopping bleeding and reducing the bleeding frequency of a hemophilia A piglet but was blocked by the inhibitor against human factor VIII. These data suggest that the hemophilia A pig is a severe hemophilia A animal model for studying not only hemophilia A gene therapy but also the next generation recombinant coagulation factors, such as recombinant factor VIII variants with a slower clearance rate.

  6. Porcine model of hemophilia A.

    Directory of Open Access Journals (Sweden)

    Yuji Kashiwakura

    Full Text Available Hemophilia A is a common X chromosome-linked genetic bleeding disorder caused by abnormalities in the coagulation factor VIII gene (F8. Hemophilia A patients suffer from a bleeding diathesis, such as life-threatening bleeding in the brain and harmful bleeding in joints and muscles. Because it could potentially be cured by gene therapy, subhuman animal models have been sought. Current mouse hemophilia A models generated by gene targeting of the F8 have difficulties to extrapolate human disease due to differences in the coagulation and immune systems between mice and humans. Here, we generated a porcine model of hemophilia A by nuclear transfer cloning from F8-targeted fibroblasts. The hemophilia A pigs showed a severe bleeding tendency upon birth, similar to human severe hemophiliacs, but in contrast to hemophilia A mice which rarely bleed under standard breed conditions. Infusion of human factor VIII was effective in stopping bleeding and reducing the bleeding frequency of a hemophilia A piglet but was blocked by the inhibitor against human factor VIII. These data suggest that the hemophilia A pig is a severe hemophilia A animal model for studying not only hemophilia A gene therapy but also the next generation recombinant coagulation factors, such as recombinant factor VIII variants with a slower clearance rate.

  7. Evolution of viral virulence: empirical studies

    Science.gov (United States)

    Kurath, Gael; Wargo, Andrew R.

    2016-01-01

    The concept of virulence as a pathogen trait that can evolve in response to selection has led to a large body of virulence evolution theory developed in the 1980-1990s. Various aspects of this theory predict increased or decreased virulence in response to a complex array of selection pressures including mode of transmission, changes in host, mixed infection, vector-borne transmission, environmental changes, host vaccination, host resistance, and co-evolution of virus and host. A fundamental concept is prediction of trade-offs between the costs and benefits associated with higher virulence, leading to selection of optimal virulence levels. Through a combination of observational and experimental studies, including experimental evolution of viruses during serial passage, many of these predictions have now been explored in systems ranging from bacteriophage to viruses of plants, invertebrates, and vertebrate hosts. This chapter summarizes empirical studies of viral virulence evolution in numerous diverse systems, including the classic models myxomavirus in rabbits, Marek's disease virus in chickens, and HIV in humans. Collectively these studies support some aspects of virulence evolution theory, suggest modifications for other aspects, and show that predictions may apply in some virus:host interactions but not in others. Finally, we consider how virulence evolution theory applies to disease management in the field.

  8. Anaerobiosis induced virulence of Salmonella typhi

    DEFF Research Database (Denmark)

    Kapoor, Sarika; Singh, R D; Sharma, P C

    2002-01-01

    BACKGROUND & OBJECTIVES: Anaerobic conditions are frequently encountered by pathogens invading the gastrointestinal tract due to low/limiting oxygen conditions prevalent in the small intestine. This anaerobic stress has been suggested to enhance the virulence of gut pathogens. In the present stud...... dismutase (SOD) and catalase. INTERPRETATION & CONCLUSION: Our results suggest that exposure of S. Typhi to anaerobic conditions enhances its virulence....

  9. THE EFFECT OF GREEN TEA EXTRACT - EPIGALLOCATECHIN GALLATE (EGCG ON PORCINE OVARIAN GRANULOSA CELL

    Directory of Open Access Journals (Sweden)

    Attila Kádasi

    2014-02-01

    Full Text Available The aim of our study was to elucidate the potential effect of green tea substance on basic ovarian functions. For this purpose, we examined the action of green tea bioactive molecule, epigallocatechin gallate (given at doses 0, 1, 10, 100 μg/mL, on cultured porcine ovarian granulosa cell functions - proliferation, apoptosis and steroidogenesis. Accumulation of PCNA (marker of proliferation, BAX (marker of apoptosis and the release of steroid hormones (progesterone and testosterone were analysed by immunocytochemistry and RIA respectively. It was observed that epigallocatechin gallate addition decreased the percentage of proliferative (PCNA-positive cells at all used doses (1, 10 and 100 μg/mL. The percentage of apoptotic (BAX-positive cells was increased at the highest used dose (100 μg/mL, but not a lower doses. Epigallocatechin gallate stimulated progesterone release (at 10 μg/mL but not at 1 and 100 μg/mL and diminished testosterone release (at 1 μg/mL but not at 10 and 100 μg/mL by porcine granulosa cells. Our results suggest a direct effect of epigallocatechin gallate on proliferation, apoptosis and steroidogenesis in porcine ovaries. Taken together, these data suggest that green tea molecule epigallocatechin gallate can negatively affect reproductive (ovarian functions – suppress ovarian cell proliferation, promote their apoptosis and alter release of steroid hormones.

  10. Establishment and characterization of a differentiated epithelial cell culture model derived from the porcine cervix uteri

    Directory of Open Access Journals (Sweden)

    Miessen Katrin

    2012-03-01

    Full Text Available Abstract Background Cervical uterine epithelial cells maintain a physiological and pathogen-free milieu in the female mammalian reproductive tract and are involved in sperm-epithelium interaction. Easily accessible, differentiated model systems of the cervical epithelium are not yet available to elucidate the underlying molecular mechanisms within these highly specialized cells. Therefore, the aim of the study was to establish a cell culture of the porcine cervical epithelium representing in vivo-like properties of the tissue. Results We tested different isolation methods and culture conditions and validated purity of the cultured cells by immunohistochemistry against keratins. We could reproducibly culture pure epithelial cells from cervical tissue explants. Based on a morphology score and the WST-1 Proliferation Assay, we optimized the growth medium composition. Primary porcine cervical cells performed best in conditioned Ham's F-12, containing 10% FCS, EGF and insulin. After cultivation in an air-liquid interface for three weeks, the cells showed a discontinuously multilayered phenotype. Finally, differentiation was validated via immunohistochemistry against beta catenin. Mucopolysaccharide production could be shown via alcian blue staining. Conclusions We provide the first suitable protocol to establish a differentiated porcine epithelial model of the cervix uteri, based on easily accessible cells using slaughterhouse material.

  11. Virulence of Fusarium species to alfalfa seedlings

    Directory of Open Access Journals (Sweden)

    Krnjaja Vesna

    2005-01-01

    Full Text Available In in vitro conditions, virulence of 91 isolates of species Fusarium genus (F. oxysporum, F. solani, F. acuminatum, F. equiseti, F. arthrosporioides, F. prolifera- tum, F. avenaceum, F. semitectum, F. tricinctum, F. sporotrichioides and F. graminearum towards alfalfa seedlings was investigated. Isolates of investigated species originated from diseased alfalfa plants collected at four locations in Serbia based on symptoms of wilting caused by Fusarium and root rotting. Pathogenicity and virulence of investigated isolates of Fusarium spp. were determined by visual evaluation of inoculated seedlings of cultivar K28 in laboratory conditions. All isolated of investigated species had pathogenic effect on alfalfa seedlings which expressed symptoms such as necrosis of root, moist rotting and "melting of seedlings". Colour of necrotic root tissue varied from light brown, brown lipstick red to explicit black, depending on the Fusarium species. Strong virulence was established in 48 isolates, medium virulence in 31 and weak virulence in 12 isolates.

  12. Virulence of Fusarium species to alfalfa seedlings

    Directory of Open Access Journals (Sweden)

    Krnjaja Vesna

    2005-01-01

    Full Text Available In in vitro conditions, virulence of 91 isolates of species Fusarium genus (F. oxysporum, F. solani, F. acuminatum, F. equiseti, F. arthrosporioides, F. proliferatum, F. avenaceum, F. semitectum, F. tricinctum, F. sporotrichioides and F. graminearum towards alfalfa seedlings was investigated. Isolates of investigated species originated from diseased alfalfa plants collected on four locations in Serbia based on symptoms of wilting caused by fusarium and root rotting. Pathogenicity and virulence of investigated isolates of Fusarium spp. were determined by visual evaluation of inoculated seedlings of cultivars K28 in laboratory conditions. All isolated of investigated species had pathogenic effect on alfalfa seedlings, which expressed symptoms such as necrosis of root, moist rotting and "melting of seedlings". Colour of necrotic root tissue varied from light brown, brown, lipstick red to explicit black, depending on the Fusarium species. Strong virulence was established in 48 isolates, medium virulence in 31 and weak virulence in 12 isolates.

  13. Identification and Characterization of IgdE, a Novel IgG-degrading Protease of Streptococcus suis with Unique Specificity for Porcine IgG*

    Science.gov (United States)

    Spoerry, Christian; Seele, Jana; Valentin-Weigand, Peter; Baums, Christoph G.; von Pawel-Rammingen, Ulrich

    2016-01-01

    Streptococcus suis is a major endemic pathogen of pigs causing meningitis, arthritis, and other diseases. Zoonotic S. suis infections are emerging in humans causing similar pathologies as well as severe conditions such as toxic shock-like syndrome. Recently, we discovered an IdeS family protease of S. suis that exclusively cleaves porcine IgM and represents the first virulence factor described, linking S. suis to pigs as their natural host. Here we report the identification and characterization of a novel, unrelated protease of S. suis that exclusively targets porcine IgG. This enzyme, designated IgdE for immunoglobulin G-degrading enzyme of S. suis, is a cysteine protease distinct from previous characterized streptococcal immunoglobulin degrading proteases of the IdeS family and mediates efficient cleavage of the hinge region of porcine IgG with a high degree of specificity. The findings that all S. suis strains investigated possess the IgG proteolytic activity and that piglet serum samples contain specific antibodies against IgdE strongly indicate that the protease is expressed in vivo during infection and represents a novel and putative important bacterial virulence/colonization determinant, and a thus potential therapeutic target. PMID:26861873

  14. Proteomic Characterization of Yersinia pestis Virulence

    Energy Technology Data Exchange (ETDEWEB)

    Chromy, B; Murphy, G; Gonzales, A; Fitch, J P; McCutchen-Maloney, S L

    2005-01-05

    Yersinia pestis, the etiological agent of plague, functions via the Type III secretion mechanism whereby virulence factors are induced upon interactions with a mammalian host. Here, the Y. pestis proteome was studied by two-dimensional differential gel electrophoresis (2-D DIGE) under physiologically relevant growth conditions mimicking the calcium concentrations and temperatures that the pathogen would encounter in the flea vector and upon interaction with the mammalian host. Over 4100 individual protein spots were detected of which hundreds were differentially expressed in the entire comparative experiment. A total of 43 proteins that were differentially expressed between the vector and host growth conditions were identified by mass spectrometry. Expected differences in expression were observed for several known virulence factors including catalase-peroxidase (KatY), murine toxin (Ymt), plasminogen activator (Pla), and F1 capsule antigen (Caf1), as well as putative virulence factors. Chaperone proteins and signaling molecules hypothesized to be involved in virulence due to their role in Type III secretion were also identified. Other differentially expressed proteins not previously reported to contribute to virulence are candidates for more detailed mechanistic studies, representing potential new virulence determinants. For example, several sugar metabolism proteins were differentially regulated in response to lower calcium and higher temperature, suggesting these proteins, while not directly connected to virulence, either represent a metabolic switch for survival in the host environment or may facilitate production of virulence factors. Results presented here contribute to a more thorough understanding of the virulence mechanism of Y. pestis through proteomic characterization of the pathogen under induced virulence.

  15. Oxidative stress induced by zearalenone in porcine granulosa cells and its rescue by curcumin in vitro.

    Directory of Open Access Journals (Sweden)

    Xunsi Qin

    Full Text Available Oxidative stress (OS, as a signal of aberrant intracellular mechanisms, plays key roles in maintaining homeostasis for organisms. The occurrence of OS due to the disorder of normal cellular redox balance indicates the overproduction of reactive oxygen species (ROS and/or deficiency of antioxidants. Once the balance is broken down, repression of oxidative stress is one of the most effective ways to alleviate it. Ongoing studies provide remarkable evidence that oxidative stress is involved in reproductive toxicity induced by various stimuli, such as environmental toxicants and food toxicity. Zearalenone (ZEA, as a toxic compound existing in contaminated food products, is found to induce mycotoxicosis that has a significant impact on the reproduction of domestic animals, especially pigs. However, there is no information about how ROS and oxidative stress is involved in the influence of ZEA on porcine granulosa cells, or whether the stress can be rescued by curcumin. In this study, ZEA-induced effect on porcine granulosa cells was investigated at low concentrations (15 μM, 30 μM and 60 μM. In vitro ROS levels, the mRNA level and activity of superoxide dismutase, glutathione peroxidase and catalase were obtained. The results showed that in comparison with negative control, ZEA increased oxidative stress with higher ROS levels, reduced the expression and activity of antioxidative enzymes, increased the intensity of fluorogenic probes 2', 7'-Dichlorodihydrofluorescin diacetate and dihydroethidium in flow cytometry assay and fluorescence microscopy. Meanwhile, the activity of glutathione (GSH did not change obviously following 60 μM ZEA treatment. Furthermore, the underlying protective mechanisms of curcumin on the ZEA-treated porcine granulosa cells were investigated. The data revealed that curcumin pre-treatment significantly suppressed ZEA-induced oxidative stress. Collectively, porcine granulosa cells were sensitive to ZEA, which may induce

  16. Simplified cryopreservation of porcine cloned blastocysts

    DEFF Research Database (Denmark)

    Du, Yutao; Zhang, Yunhai; Li, Juan

    2007-01-01

    )â€"handmade cloning (HMC)â€"to establish a simplified and efficient cryopreservation system for porcine cloned embryos. In Experiment 1, zonae pellucidae of oocytes were partially digested with pronase, followed by centrifugation to polarize lipid particles. Ninety percent (173/192) oocytes were successfully......). Our results prove that porcine embryos produced from delipated oocytes by PA or HMC can be cryopreserved effectively by ultrarapid vitrification. Further experiments are required to assess the in vivo developmental competence of the cloned-vitrified embryos  ...

  17. Susceptibility of human liver cells to porcine endogenous retrovirus.

    Science.gov (United States)

    Lin, Xinzi; Qi, Lin; Li, Zhiguo; Chi, Hao; Lin, Wanjun; Wang, Yan; Jiang, Zesheng; Pan, Mingxin; Gao, Yi

    2013-12-01

    The risk of porcine endogenous retrovirus infection is a major barrier for pig-to-human xenotransplant. Porcine endogenous retrovirus, present in porcine cells, can infect many human and nonhuman primate cells in vitro, but there is no evidence available about in vitro infection of human liver cells. We investigated the susceptibility of different human liver cells to porcine endogenous retrovirus. The supernatant from a porcine kidney cell line was added to human liver cells, including a normal hepatocyte cell line (HL-7702 cells), primary hepatocytes (Phh cells), and a liver stellate cell line (Lx-2 cells), and to human embryonic kidney cells as a reference control. Expression of the porcine endogenous retrovirus antigen p15E in the human cells was evaluated with polymerase chain reaction, reverse transcription-polymerase chain reaction, and Western blot. The porcine endogenous retrovirus antigen p15E was not expressed in any human liver cells (HL-7702, Phh, or Lx-2 cells) that had been exposed to supernatants from porcine kidney cell lines. Porcine endogenous retrovirus-specific fragments were amplified in human kidney cells. Human liver cells tested were not susceptible to infection by porcine endogenous retrovirus. Therefore, not all human cells are susceptible to porcine endogenous retrovirus.

  18. Rapid detection of porcine circovirus type 2 using a TaqMan-based real-time PCR

    Directory of Open Access Journals (Sweden)

    Zhang Chunling

    2010-12-01

    Full Text Available Abstract Porcine circovirus type 2 (PCV2 and the associated disease postweaning multisystemic wasting syndrome (PMWS have caused heavy losses in global agriculture in recent decades. Rapid detection of PCV2 is very important for the effective prophylaxis and treatment of PMWS. To establish a sensitive, specific assay for the detection and quantitation of PCV2, we designed and synthesized specific primers and a probe in the open reading frame 2. The assay had a wide dynamic range with excellent linearity and reliable reproducibility, and detected between 102 and 1010 copies of the genomic DNA per reaction. The coefficient of variation for Ct values varied from 0.59% to 1.05% in the same assay and from 1.9% to 4.2% in 10 different assays. The assay did not cross-react with porcine circovirus type 1, porcine reproductive and respiratory, porcine epidemic diarrhea, transmissible gastroenteritis of pigs and rotavirus. The limits of detection and quantitation were 10 and 100 copies, respectively. Using the established real-time PCR system, 39 of the 40 samples we tested were detected as positive.

  19. Virulence Factors of A Review

    Directory of Open Access Journals (Sweden)

    Bruna M. Roesler

    2014-07-01

    Full Text Available Helicobacter pylori is a spiral-shaped Gram-negative bacterium that colonizes the human stomach and can establish a long-term infection of the gastric mucosa, a condition that affects the relative risk of developing various clinical disorders of the upper gastrointestinal tract, such as chronic gastritis, peptic ulcer disease, mucosa-associated lymphoid tissue (MALT lymphoma, and gastric adenocarcinoma. H. pylori presents a high-level of genetic diversity, which can be an important factor in its adaptation to the host stomach and also for the clinical outcome of infection. There are important H. pylori virulence factors that, along with host characteristics and the external environment, have been associated with the different occurrences of diseases. This review is aimed to analyzing and summarizing the main of them and possible associations with the clinical outcome.

  20. Interlaboratory testing of porcine sera for antibodies to porcine circovirus type 2

    DEFF Research Database (Denmark)

    McNair, I.; Marshall, M.; McNeilly, F.

    2004-01-01

    A panel of 20 porcine sera was distributed to 5 laboratories across Europe and Canada. Each center was requested to test the sera for the presence of porcine circovirus type 2 antibodies using the routine assays, indirect immunofluorescence assay (IFA) and indirect immunoperoxidase monolayer assa...... than did IFA, and paraformaldehyde gave higher titers than did acetone or ethyl alcohol. This report highlights the need for standardized procedures and biologicals for this virus....

  1. Pyocyanin inhibits both nitric oxide-dependent and -independent relaxation in porcine coronary arteries.

    Science.gov (United States)

    Hempenstall, Allison; Grant, Gary D; Anoopkumar-Dukie, Shailendra; Johnson, Peter J

    2015-02-01

    The effects of the Pseudomonas aeruginosa virulence factor pyocyanin (PCN) on the contractile function of porcine coronary arteries was investigated in vitro. Artery rings (5 mm) were suspended in organ baths containing Krebs' solution for the measurement of isometric tension. The effect of PCN on resting and precontracted coronary arteries was initially investigated with various agents. Arteries were precontracted with prostaglandin (PG) F2α or potassium chloride and endothelium-dependent relaxations were induced by various agents in the presence of PCN. Pyocyanin (0.1-10 μmol/L) evoked small-amplitude, dose-dependent contractions in resting porcine coronary arteries. In addition, PCN amplified the contractile response to PGF2α , but did not alter responses to carbachol. Pyocyanin (0.1-10 μmol/L) significantly inhibited endothelium-dependent relaxations evoked by neurokinin A. Pyocyanin also inhibited relaxations evoked by diethylamine nitric oxide (a nitric oxide donor), forskolin (an adenylate cyclase activator), dibuytyryl-cAMP (a cAMP analogue), 8-bromo-cGMP (a cGMP analogue) and P1075 (a KATP channel activator), but not isoprenaline (β-adrenoceceptor agonist). These results indicate that physiological concentrations of PCN interfere with multiple intracellular processes involved in vascular smooth muscle relaxation, in particular pathways downstream of nitric oxide release. Thus, PCN may alter normal vascular function in patients infected with P. aeruginosa. © 2014 Wiley Publishing Asia Pty Ltd.

  2. Distribution of virulence plasmids within Salmonellae.

    Science.gov (United States)

    Woodward, M J; McLaren, I; Wray, C

    1989-03-01

    The virulence region of the Salmonella dublin 50 MDa plasmid shared homology with 678 of 1021 salmonellae tested in colony hybridization experiments. The majority of S. dublin, S. typhimurium and S. enteritidis isolates tested hybridized with the region whereas, with the exception of S. hessarek, S. pullorum and S. gallinarum, other serotypes did not. Homologous virulence regions were plasmid encoded. In S. typhimurium a common 60 MDa plasmid was present in all phage types tested but not in DT4, DT37 and DT170. Smaller plasmids showing partial homology were found in DT12, DT18, DT193 and DT204C. In S. enteritidis a distinct plasmid profile for each of eight phage types was observed. Hybridizing plasmids were found in DT3, DT4, DT8, DT9 and DT11 whereas DT7, which was plasmid free, and DT10 and DT14, which harboured plasmids, did not hybridize. The extent of homology shared between S. dublin, S. typhimurium and S. enteritidis virulence plasmids was about 10 MDa and appeared conserved. Virulence plasmids from S. typhimurium and S. enteritidis did not show homology with a region of the S. dublin 50 MDa plasmid which was not associated with virulence functions whereas plasmids of about 24 MDa and 38 MDa in some S. typhimurium phage types did. The association of conserved virulence regions upon differing plasmids within salmonellae is discussed with reference to possible mechanisms of distribution and evolution of virulence genes.

  3. Female reproductive disorders

    DEFF Research Database (Denmark)

    Crain, D Andrew; Janssen, Sarah J; Edwards, Thea M

    2008-01-01

    To evaluate the possible role of endocrine-disrupting compounds (EDCs) on female reproductive disorders emphasizing developmental plasticity and the complexity of endocrine-dependent ontogeny of reproductive organs. Declining conception rates and the high incidence of female reproductive...... disruptions warrant evaluation of the impact of EDCs on female reproductive health....

  4. Reproductive Medicine in Amphibians.

    Science.gov (United States)

    Chai, Norin

    2017-05-01

    Reproduction of amphibians includes ovulation, spermiation, fertilization, oviposition, larval stage and development, and metamorphosis. A problem at any stage could lead to reproductive failure. To stimulate reproduction, environmental conditions must be arranged to simulate changes in natural habits. Reproductive life history is well documented in amphibians; a thorough knowledge of this subject will aid the practitioner in diagnosis and treatment. Technologies for artificial reproduction are developing rapidly, and some protocols may be transferable to privately kept or endangered species. Reproductive tract disorders are rarely described; no bacterial or viral diseases are known that specifically target the amphibian reproductive system. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. The Porcine TSPY Gene Is Tricopy but Not a Copy Number Variant.

    Directory of Open Access Journals (Sweden)

    Anh T Quach

    Full Text Available The testis-specific protein Y-encoded (TSPY gene is situated on the mammalian Y-chromosome and exhibits some remarkable biological characteristics. It has the highest known copy number (CN of all protein coding genes in the human and bovine genomes (up to 74 and 200, respectively and also shows high individual variability. Although the biological function of TSPY has not yet been elucidated, its specific expression in the testis and several identified binding domains within the protein suggests roles in male reproduction. Here we describe the porcine TSPY, as a multicopy gene with three copies located on the short arm of the Y-chromosome with no variation at three exon loci among 20 animals of normal reproductive health from four breeds of domestic pigs (Piétrain, Landrace, Duroc and Yorkshire. To further investigate the speculation that porcine TSPY is not a copy number variant, we have included five Low-fertility boars and five boars with exceptional High-fertility records. Interestingly, there was no difference between the High- and Low-fertile groups, but we detected slightly lower TSPY CN at all three exons (2.56-2.85 in both groups, as compared to normal animals, which could be attributed to technical variability or somatic mosaicism. The results are based on both relative quantitative real-time PCR (qPCR and droplet digital PCR (ddPCR. Chromosomal localization of the porcine TSPY was done using fluorescence in situ hybridization (FISH with gene specific PCR probes.

  6. Experimentally induced Porcine Coccidiosis | Onawunmi | Nigerian ...

    African Journals Online (AJOL)

    Journal Home > Vol 4, No 2 (1977) >. Log in or Register to get access to full text downloads. Username, Password, Remember me, or Register. DOWNLOAD FULL TEXT Open Access DOWNLOAD FULL TEXT Subscription or Fee Access. Experimentally induced Porcine Coccidiosis. OA Onawunmi. Abstract. No abstract.

  7. porcine anaesthesia for advanced trauma operative management

    African Journals Online (AJOL)

    David Ofori-Adjei

    2008-09-01

    Sep 1, 2008 ... 120. PORCINE ANAESTHESIA FOR ADVANCED TRAUMA OPERATIVE. MANAGEMENT (ATOM). H. BADDOO, F. AHIAKU, E. FORDJUOR, I. WULFF, D. AKUOKU and D. KWAMI. Department of Anaesthesia, University of Ghana Medical School, Accra, Ghana. Author for correspondence: Dr Henry Baddoo.

  8. Short communication Expression profiling analyses of porcine ...

    African Journals Online (AJOL)

    p2492989

    (ORF) of the porcine MuRF1 gene consisting of 354 amino acids was obtained and it shared 93% and 90% identity with those of ... Real-time quantitative reverse transcriptase-polymerase chain reaction ... identified as one of the members of the E3 ubiquitin ligases and it regulates myofibrillar protein metabolism. (Bodine et ...

  9. Porcine Circovirus Diseases: A review of PMWS

    DEFF Research Database (Denmark)

    Baekbo, P.; Kristensen, C. S.; Larsen, L. E.

    2012-01-01

    Porcine Circo Virus type 2 have been coming on the market and many studies have shown great benefits of these to control PMWS. Today, sow vaccines as well as piglet vaccines are available in most countries. An extensive meta‐analysis of many of the vaccines has shown a comparable good efficacy...

  10. Mapping markers linked to porcine salmonellosis susceptibility

    NARCIS (Netherlands)

    Galina-Pantoja, L.; Siggens, K.; Schriek, M.G.; Heuven, H.C.M.|info:eu-repo/dai/nl/314417818

    2009-01-01

    Anim Genet. 2009 Jun 3. [Epub ahead of print] Mapping markers linked to porcine salmonellosis susceptibility. Galina-Pantoja L, Siggens K, van Schriek MG, Heuven HC. PIC/Genus, 100 Bluegrass Commons Blvd, Hendersonville, TN 37075, USA. The goal of this study was to identify pig chromosomal regions

  11. Porcine Tricuspid Valve Anatomy and Human Compatibility

    DEFF Research Database (Denmark)

    Waziri, Farhad; Lyager Nielsen, Sten; Hasenkam, J. Michael

    2016-01-01

    before clinical use. The study aim was to evaluate and compare the tricuspid valve anatomy of porcine and human hearts. METHODS: The anatomy of the tricuspid valve and the surrounding structures that affect the valve during a cardiac cycle were examined in detail in 100 fresh and 19 formalin...

  12. The extinction differential induced virulence macroevolution

    Science.gov (United States)

    Zhang, Feng; Xu, Liufang; Wang, Jin

    2014-04-01

    We apply the potential-flux landscape theory to deal with the large fluctuation induced extinction phenomena. We quantify the most probable extinction pathway on the landscape and measure the extinction risk by the landscape topography. In this Letter, we investigate the disease extinction through an epidemic model described by a set of chemical reaction. We found the virulence-differential-dependent symbioses between mother and daughter pathogen species: mutualism and parasitism. The symbioses, whether mutualism or parasitism, benefit the higher virulence species. This implies that speciation towards lower virulence is an effective strategy for a pathogen species to reduce its extinction risk.

  13. Reproductive Disorders in Snakes.

    Science.gov (United States)

    Di Girolamo, Nicola; Selleri, Paolo

    2017-05-01

    Reproduction of snakes is one of the challenging aspects of herpetology medicine. Due to the complexity of reproduction, several disorders may present before, during, or after this process. This article describes the physical examination, and radiographic, ultrasonographic, and endoscopic findings associated with reproductive disorders in snakes. Surgical techniques used to resolve reproductive disorders in snakes are described. Finally, common reproductive disorders in snakes are individually discussed. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Validation and aplication of a polymerase chain reaction (PCR to detect porcine circovirus type 2 (PCV-2 in swine sera

    Directory of Open Access Journals (Sweden)

    Luisa Fernanda Villadiego Marmolejo

    2007-12-01

    Full Text Available Porcine circovirosis is an infectious-contagious syndrome caused by porcine circovirus type 2 (PCV-2 found mainly in recently weaned piglets causing dermatitis, neurological and reproductive disorders, pneumonia and encephalitis. The objectives of the present study were to validate a Polymerase Chain Reaction (PCR technique to detect PCV-2 in swine serum and to apply the validated technique in swine serum samples to detect PCV-2. After the application of two different PCRs, 100% of the surveyed animals were negative to PCV-2; furthermore, the PCR targeted to a region between ORFs 1 and 2 of the virus was found more sensitive when compared to another PCR targeted to the capsid protein gene. As a conclusion, PCR is a valid technique to detect PCV-2 in swine serum and the surveyed population was free of the virus.

  15. Full-length genomic analysis of Korean porcine Sapelovirus strains.

    Science.gov (United States)

    Son, Kyu-Yeol; Kim, Deok-Song; Kwon, Joseph; Choi, Jong-Soon; Kang, Mun-Il; Belsham, Graham J; Cho, Kyoung-Oh

    2014-01-01

    Porcine sapelovirus (PSV), a species of the genus Sapelovirus within the family Picornaviridae, is associated with diarrhea, pneumonia, severe neurological disorders, and reproductive failure in pigs. However, the structural features of the complete PSV genome remain largely unknown. To analyze the structural features of PSV genomes, the full-length nucleotide sequences of three Korean PSV strains were determined and analyzed using bioinformatic techniques in comparison with other known PSV strains. The Korean PSV genomes ranged from 7,542 to 7,566 nucleotides excluding the 3' poly(A) tail, and showed the typical picornavirus genome organization; 5'untranslated region (UTR)-L-VP4-VP2-VP3-VP1-2A-2B-2C-3A-3B-3C-3D-3'UTR. Three distinct cis-active RNA elements, the internal ribosome entry site (IRES) in the 5'UTR, a cis-replication element (CRE) in the 2C coding region and 3'UTR were identified and their structures were predicted. Interestingly, the structural features of the CRE and 3'UTR were different between PSV strains. The availability of these first complete genome sequences for PSV strains will facilitate future investigations of the molecular pathogenesis and evolutionary characteristics of PSV.

  16. Biochemical Basis of Virulence in Epidemic Typhus.

    Science.gov (United States)

    1983-01-01

    properties include antibiotic resistance, toxin production , fertility, bacteriocin production , and production of virulence factors. (4). Numerous...sample. The rickettsial leukotoxin was probably not a soluble product , was active in the absence of phagocytosis, and was inhibited by inactivation of

  17. Virulence Factors IN Fungi OF Systemic Mycoses

    Directory of Open Access Journals (Sweden)

    KUROKAWA Cilmery Suemi

    1998-01-01

    Full Text Available Pathogenic fungi that cause systemic mycoses retain several factors which allow their growth in adverse conditions provided by the host, leading to the establishment of the parasitic relationship and contributing to disease development. These factors are known as virulence factors which favor the infection process and the pathogenesis of the mycoses. The present study evaluates the virulence factors of pathogenic fungi such as Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum and Paracoccidioides brasiliensis in terms of thermotolerance, dimorphism, capsule or cell wall components as well as enzyme production. Virulence factors favor fungal adhesion, colonization, dissemination and the ability to survive in hostile environments and elude the immune response mechanisms of the host. Both the virulence factors presented by different fungi and the defense mechanisms provided by the host require action and interaction of complex processes whose knowledge allows a better understanding of the pathogenesis of systemic mycoses.

  18. Structural Genomics of Bacterial Virulence Factors

    Science.gov (United States)

    2006-05-01

    plant proteins. Its presence in the virulence-related pXO1 plasmid of Bacillus anthracis (pX01-01) as well as in several other pathogens makes it a...from several other bacilli; Enterococcus, Listeria , Lactococcus, Lactobacillus, or other Bacillus species. Function of proteins from this family is...virulence plasmids. Infect Immun 71: 2736-2743. Braun, L. and P. Cossart. 2000. Interactions between Listeria monocytogenes and host mammalian cells

  19. E. coli Nissle 1917 Affects Salmonella adhesion to porcine intestinal epithelial cells.

    Directory of Open Access Journals (Sweden)

    Peter Schierack

    Full Text Available BACKGROUND: The probiotic Escherichia coli strain Nissle 1917 (EcN has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we investigated the inhibitory effects of EcN on Salmonella Typhimurium invasion of porcine intestinal epithelial cells, focusing on EcN effects on the various stages of Salmonella infection including intracellular and extracellular Salmonella growth rates, virulence gene regulation, and adhesion. We show that EcN affects the initial Salmonella invasion steps by modulating Salmonella virulence gene regulation and Salmonella SiiE-mediated adhesion, but not extra- and intracellular Salmonella growth. However, the inhibitory activity of EcN against Salmonella invasion always correlated with EcN adhesion capacities. EcN mutants defective in the expression of F1C fimbriae and flagellae were less adherent and less inhibitory toward Salmonella invasion. Another E. coli strain expressing F1C fimbriae was also adherent to IPEC-J2 cells, and was similarly inhibitory against Salmonella invasion like EcN. CONCLUSIONS: We propose that EcN affects Salmonella adhesion through secretory components. This mechanism appears to be common to many E. coli strains, with strong adherence being a prerequisite for an effective reduction of SiiE-mediated Salmonella adhesion.

  20. Preclinical detection of porcine circovirus type 2 infection using an ultrasensitive nanoparticle DNA probe-based PCR assay.

    Directory of Open Access Journals (Sweden)

    Yong Huang

    Full Text Available Porcine circovirus type 2 (PCV2 has emerged as one of the most important pathogens affecting swine production globally. Preclinical identification of PCV2 is very important for effective prophylaxis of PCV2-associated diseases. In this study, we developed an ultrasensitive nanoparticle DNA probe-based PCR assay (UNDP-PCR for PCV2 detection. Magnetic microparticles coated with PCV2 specific DNA probes were used to enrich PCV2 DNA from samples, then gold nanoparticles coated with PCV2 specific oligonucleotides were added to form a sandwich nucleic acid-complex. After the complex was formed, the oligonucleotides were released and characterized by PCR. This assay exhibited about 500-fold more sensitive than conventional PCR, with a detection limit of 2 copies of purified PCV2 genomic DNA and 10 viral copies of PCV2 in serum. The assay has a wide detection range for all of PCV2 genotypes with reliable reproducibility. No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 1, porcine parvovirus, porcine pseudorabies virus, porcine reproductive and respiratory syndrome virus and classical swine fever virus. The positive detection rate of PCV2 specific UNDP-PCR in 40 preclinical field samples was 27.5%, which appeared greater than that by conventional and real-time PCR and appeared application potency in evaluation of the viral loads levels of preclinical infection samples. The UNDP-PCR assay reported here can reliably rule out false negative results from antibody-based assays, provide a nucleic acid extraction free, specific, ultrasensitive, economic and rapid diagnosis method for preclinical PCV2 infection in field, which may help prevent large-scale outbreaks.

  1. Morphological assessment of sucrose preservation for porcine heart valves.

    OpenAIRE

    Drury, P J; Olsen, E G; Ross, D N

    1982-01-01

    Porcine aortic valves stored in various concentrations of sucrose (50-80%) for up to 52 weeks were examined both histologically and by electron microscopy. The valves were compared with porcine aortic valves stored in a nutrient and antibiotic medium for 12 weeks. Overall preservation was better in those porcine valves stored in sucrose solution than in nutrient and antibiotic medium, the best preservation being in 50% sucrose. Despite wide separation of collagen at that concentration seen on...

  2. Comparison of high and low virulence serotypes of Actinobacillus pleuropneumoniae by quantitative real-time PCR

    DEFF Research Database (Denmark)

    Schou, Kirstine Klitgaard; Angen, Øystein; Boye, Mette

    of high virulence while serotype 6 strains are normally found to be less pathogenic. To gain an understanding of the differential virulence of serotype 2 and 6, the expression of a panel of Ap genes during infection of porcine epithelial lung cells (SJPL) were examined by quantitative real-time PCR (qPCR...... to be important for early establishment of the bacteria in the host were examined by qPCR. The genes examined were apfA, coding for a subunit of Type IV pili, kdsB coding for a gene involved in lippopolysacceride biosynthesis, and pgaB which is involved in biofilm formation, all three believed to be important...... with respect to host cells adhesion. Also included in the analysis were the capsular gene, cpxB, the RTX toxin genes apxII, and apxIV and the gene exbD, involved in binding of iron from host cells. Finally, three previously validated reference genes, glyA, pykA and tpiA were included for normalization of the qPCR...

  3. Monoclonal antibodies specific to heat-treated porcine blood.

    Science.gov (United States)

    Raja Nhari, Raja Mohd Hafidz; Hamid, Muhajir; Rasli, Nurmunirah Mohamad; Omar, Abdul Rahman; El Sheikha, Aly Farag; Mustafa, Shuhaimi

    2016-05-01

    Porcine blood is potentially being utilized in food as a binder, gelling agent, emulsifier or colorant. However, for certain communities, the usage of animal blood in food is strictly prohibited owing to religious concerns and health reasons. This study reports the development of monoclonal antibodies (MAbs) against heat-treated soluble proteins (HSPs) of autoclaved porcine blood; characterization of MAbs against blood, non-blood and plasma from different animal species using qualitative indirect non-competitive enzyme-linked immunosorbent assay (ELISA); and immunoblotting of antigenic components in HSPs of porcine blood. Fifteen MAbs are specific to heat-treated and raw porcine blood and not cross-reacted with other animal blood and non-blood proteins (meat and non-meat). Twelve MAbs are specific to porcine plasma, while three MAbs specific to porcine plasma are cross-reacted with chicken plasma. Immunoblotting revealed antigenic protein bands (∼60, ∼85-100 and ∼250 kDa) in porcine blood and plasma recognized by the MAbs. Selection of MAbs that recognized 60 kDa HSPs of porcine blood and plasma as novel monoclonal antibodies would be useful for detection of porcine plasma in processed food using the immunoassay method. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  4. Detection of porcine circovirus genotypes 2a and 2b in aborted foetuses from infected swine herds in the State of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    de Castro Alessandra MMG

    2012-05-01

    Full Text Available Abstract Background Porcine circovirus type 2 (PCV2 has been associated with several disease complexes, including reproductive failure. The aim of this study was to identify the subtypes of PCV2 that are associated with reproductive failure in pigs from the State of São Paulo, Brazil and to investigate co-infections with other infectious organisms. Findings Samples of 168 aborted foetuses or mummified foetuses from five farrow-to-finish swine farms known to be infected with PCV2 and located in the State of São Paulo were tested for PCV2 by polymerase chain reaction (PCR. Positive samples were additionally tested for porcine parvovirus (PPV, Leptospira spp. and Brucella spp. by PCR. PCV2 was detected in 18 of the samples (10.7%. PPV, Brucella spp. and Leptospira spp were found in 2, 10 and 0 cases, respectively. Eleven PCV2 strains were sequenced and determined to be either genotype 2a (n = 1 or 2b (n = 10. Conclusions The findings indicate that the frequency of PCV2 infections in aborted porcine foetuses from the State of São Paulo is rather low (10.7% and that co-infection with other pathogens is common and may be involved in PCV2 associated reproductive failure. No repeatable, characteristic amino acid motifs for regions of the PCV2 capsid protein seemed to be associated with abortion in sows.

  5. Identification of Secreted Exoproteome Fingerprints of Highly-Virulent and Non-Virulent Staphylococcus aureus Strains.

    Science.gov (United States)

    Bonar, Emilia; Wojcik, Iwona; Jankowska, Urszula; Kedracka-Krok, Sylwia; Bukowski, Michal; Polakowska, Klaudia; Lis, Marcin W; Kosecka-Strojek, Maja; Sabat, Artur J; Dubin, Grzegorz; Friedrich, Alexander W; Miedzobrodzki, Jacek; Dubin, Adam; Wladyka, Benedykt

    2016-01-01

    Staphylococcus aureus is a commensal inhabitant of skin and mucous membranes in nose vestibule but also an important opportunistic pathogen of humans and livestock. The extracellular proteome as a whole constitutes its major virulence determinant; however, the involvement of particular proteins is still relatively poorly understood. In this study, we compared the extracellular proteomes of poultry-derived S. aureus strains exhibiting a virulent (VIR) and non-virulent (NVIR) phenotype in a chicken embryo experimental infection model with the aim to identify proteomic signatures associated with the particular phenotypes. Despite significant heterogeneity within the analyzed proteomes, we identified alpha-haemolysin and bifunctional autolysin as indicators of virulence, whereas glutamylendopeptidase production was characteristic for non-virulent strains. Staphopain C (StpC) was identified in both the VIR and NVIR proteomes and the latter fact contradicted previous findings suggesting its involvement in virulence. By supplementing NVIR, StpC-negative strains with StpC, and comparing the virulence of parental and supplemented strains, we demonstrated that staphopain C alone does not affect staphylococcal virulence in a chicken embryo model.

  6. Targeted Porcine Genome Engineering with TALENs

    DEFF Research Database (Denmark)

    Luo, Yonglun; Lin, Lin; Golas, Mariola Monika

    2015-01-01

    Genetically modified pigs are becoming an invaluable animal model for agricultural, pharmaceutical, and biomedical applications. Unlike traditional transgenesis, which is accomplished by randomly inserting an exogenous transgene cassette into the natural chromosomal context, targeted genome editing...... confers precisely editing (e.g., mutations or indels) or insertion of a functional transgenic cassette to user-designed loci. Techniques for targeted genome engineering are growing dramatically and include, e.g., zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs......, including construction of sequence-specific TALENs, delivery of TALENs into primary porcine fibroblasts, and detection of TALEN-mediated cleavage, is described. This chapter is useful for scientists who are inexperienced with TALEN engineering of porcine cells as well as of other large animals....

  7. Splicing variants of porcine synphilin-1

    DEFF Research Database (Denmark)

    Larsen, Knud Erik; Madsen, Lone Bruhn; Farajzadeh, Leila

    2015-01-01

    %) and to mouse (84%) synphilin-1. Three shorter transcript variants of the synphilin-1 gene were identified, all lacking one or more exons. SNCAIP transcripts were detected in most examined organs and tissues and the highest expression was found in brain tissues and lung. Conserved splicing variants and a novel......RNA was investigated by RNAseq. The presented work reports the molecular cloning and characterization of the porcine (Sus scrofa) synphilin-1 cDNA (SNCAIP) and three splice variants hereof. The porcine SNCAIP cDNA codes for a protein (synphilin-1) of 919 amino acids which shows a high similarity to human (90...... splice form of synhilin-1 were found in this study. All synphilin-1 isoforms encoded by the identified transcript variants lack functional domains important for protein degradation....

  8. The effect of yucca on proliferation, apoptosis, and steroidogenesis of porcine ovarian granulosa cells

    Directory of Open Access Journals (Sweden)

    Aneta Štochmaľová

    2014-02-01

    Full Text Available Yucca shidigera is a medicinal plant native to Mexico. Is a plant widely used in folk medicine to treat a variety of ailmentary disorders, but its action on reproductive processes and possible mechanisms of such action remains unknown. Yucca schidigera extract contains a number of steroidal saponins that, because of their biological activity, have attracted attention from the food industry for many years. Yucca extract is used as a natural feed additive with positive effect to microflora, digestion, metabolism and to improve animal muscle growth. Its extract has been used as a foodstuff and folk medicine to treat a wide variety of diseases for many years. Nevertheless, it remaines unknown, whether consumption of yucca can affect reproductive system. The aim of this study was to examine the effects of yucca on basic ovarian cell functions - proliferation, apoptosis and steroidogenesis. Porcine ovarian granulosa cells were cultured with and without yucca extract (added at doses 0; 1; 10 and 100 μg.mL-1 of medium. Markers of proliferation (% of PCNA-positive cells and apoptosis (% cells containing bax were analysed by immunocytochemistry. Release of steroid hormones (progesterone and testosterone was measured by EIA. It was observed, that addition of yucca inhibited proliferation (expression of PCNA, increased apoptosis (expression of bax, stimulated progesterone and inhibited testosterone release. The ability of yucca to reduce ovarian cell proliferation, to promote ovarian cell apoptosis and affect steroidogenesis demonstrates the direct influence of yucca on female gonads. Furthermore, our observations suggest the multiple sites of action (proliferation, apoptosis, steroidogenesis of yucca on porcine ovarian cell functions. It is not to be excluded, that consumption of yucca can suppress female reproductive functions.

  9. Squalus cubensis Reproduction Data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Reproductive data from Squalus cubensis (Cuban dogfish) were opportunistically collected from 2005-2012. Data include those necessary to examine reproductive cycle,...

  10. Thermal and biomechanical parameters of porcine cornea.

    Science.gov (United States)

    Kampmeier, J; Radt, B; Birngruber, R; Brinkmann, R

    2000-05-01

    New methods in refractive surgery require a considerable understanding of the material "cornea" and are often studied by theoretical modeling in order to gain insight into the procedure and an optimized approach to the technique. The quality of these models is highly dependent on the preciseness of its input parameters. Porcine cornea often is used as a model in preclinical studies because of its similarity to man and its availability. The important physical parameters for biomechanical deformation, heat conduction, and collagen denaturation kinetics have been determined for porcine cornea. Experimental methods include densitometry, calorimetry, turbidimetry, tensile tests, stress relaxation, and hydrothermal isometric tension measurements. The density of porcine cornea was measured as p = 1062+/-5 kg/m3, the heat capacity gave c = 3.74+/-0.05 J/gK. The stress-strain relation for corneal strips is represented by a third order approximation where the secant modulus yields about Esec approximately equal to 0.4 MPa for small strains less than 2%. The normalized stress relaxation is described by an exponential fit over time. The denaturation process of cornea is characterized by specific temperatures which can be related to the change of the mechanical properties. Denaturation kinetics are described according to the model of Arrhenius yielding the activation energy deltaEa = 106 kJ/mol and the phase transition entropy deltaS = 39 J/(mol x K). The established set of parameters characterizes the porcine cornea in a reliable way that creates a basis for corneal models. It furthermore gives direct hints of how to treat cornea in certain refractive techniques.

  11. Porcine stress syndrome (PSS) in Mangalitsa pigs

    OpenAIRE

    Stanišić, N.; Aleksić, S.; Di, L.; Stanimirović, Z.; Zhenhua, G.; Petrović, M.; Delić, N.; Radović, Č.; Parunović, N.; Gogić, M.

    2012-01-01

    Porcine stress syndrome (PSS) is one kind of molecular genetics defect which will cause malignant hyperthermia syndrome in pigs. It was reported that mutation of pig rynodine receptor (RYR1) gene is the main reason for PSS. The aim of this study was to test the RYR1 genotype of 10 Mangalitsa pigs using a polymerase chain reaction-restriction endonuclease fragment length polymorphism (PCR-RFLP) technique, which is a reliable and simple method for RYR1 gene t...

  12. A proteomic approach to porcine saliva.

    Science.gov (United States)

    Gutiérrez, Ana M; Cerón, José J; Fuentes-Rubio, María; Tecles, Fernando; Beeley, Josie A

    2014-02-01

    This paper reviews recent progress in salivary animal proteomics, with special reference to the porcine proteome. Until fairly recently, most studies on saliva as a diagnostic fluid have focused on humans, primates and rodents, and the development of salivary analysis in monitoring health in farm animals including pigs has received only limited consideration. The porcine salivary proteome has been characterised by 2D-electrophoresis followed by mass spectrometry. Major and minor proteins have been identified. The use of saliva as a non-invasive biological fluid in monitoring health and disease in pigs will be reviewed, together with the potential use of proteomics for the development of biomarkers. In this review, methods of collection and the composition of porcine saliva will be considered, together with saliva handling and analysis. The overall findings indicate that there is considerable potential for the development of salivary analysis as a non-invasive diagnostic fluid in the pig, and that it offers advantages over other body fluids in this animal.

  13. Posterior repair with perforated porcine dermal graft

    Directory of Open Access Journals (Sweden)

    G. Bernard Taylor

    2008-02-01

    Full Text Available OBJECTIVE: To compare postoperative vaginal incision separation and healing in patients undergoing posterior repair with perforated porcine dermal grafts with those that received grafts without perforations. Secondarily, the tensile properties of the perforated and non-perforated grafts were measured and compared. MATERIALS AND METHODS: This was a non-randomized retrospective cohort analysis of women with stage II or greater rectoceles who underwent posterior repair with perforated and non-perforated porcine dermal grafts (PelvicolTM CR Bard Covington, GA USA. The incidence of postoperative vaginal incision separation (dehiscence was compared. A secondary analysis to assess graft tensile strength, suture pull out strength, and flexibility after perforation was performed using standard test method TM 0133 and ASTM bending and resistance protocols. RESULTS: Seventeen percent of patients (21/127 who received grafts without perforations developed vaginal incision dehiscence compared to 7% (5/71 of patients who received perforated grafts (p = 0.078. Four patients with vaginal incision dehiscence with non-perforated grafts required surgical revision to facilitate healing. Neither tensile strength or suture pull out strength were significantly different between perforated and non-perforated grafts (p = 0.81, p = 0.29, respectively. There was no difference in the flexibility of the two grafts (p = 0.20. CONCLUSION: Perforated porcine dermal grafts retain their tensile properties and are associated with fewer vaginal incision dehiscences.

  14. Unique virulence properties of Yersinia enterocolitica O:3--an emerging zoonotic pathogen using pigs as preferred reservoir host.

    Science.gov (United States)

    Valentin-Weigand, Peter; Heesemann, Jürgen; Dersch, Petra

    2014-10-01

    Enteropathogenic Yersinia enterocolitica bioserotype 4/O:3 are the most frequent cause of human yersiniosis worldwide with symptoms ranging from mild diarrhea to severe complications of mesenteric lymphadenitis, liver abscesses and postinfectious extraintestinal sequelae. The main reservoir host of 4/O:3 strains are pigs, which represent a substantial disease-causing potential for humans, as they are usually asymptomatic carriers. Y. enterocolitica O:3 initiates infections by tight attachment to the intestinal mucosa. Colonization of the digestive tract is frequently followed by invasion of the intestinal layer primarily at the follicle-associated epithelium, allowing the bacteria to propagate in the lamina propria and disseminate into deeper tissues. Molecular characterization of Y. enterocolitica O:3 isolates led to the identification of (i) alternative virulence and fitness factors and (ii) small genetic variations which cause profound changes in their virulence gene expression pattern (e.g. constitutive expression of the primary invasion factor InvA). These changes provoke a major difference in the virulence properties, i.e. reduced colonization of intestinal tissues in mice, but improved long-term colonization in the pig intestine. Y. enterocolitica O:3 strains cause also a considerably lower level of proinflammatory cytokine IL-8 and higher levels of the anti-inflammatory cytokine IL-10 in porcine primary macrophages, as compared to murine macrophages, which could contribute to limiting inflammation, immunopathology and severity of the infection in pigs. Copyright © 2014 Elsevier GmbH. All rights reserved.

  15. Male Reproductive System (For Teens)

    Science.gov (United States)

    ... Why Exercise Is Wise Are Detox Diets Safe? Male Reproductive System KidsHealth > For Teens > Male Reproductive System ... and female reproductive systems. continue What Is the Male Reproductive System? Most species have two sexes: male ...

  16. Salmonella promotes virulence by repressing cellulose production.

    Science.gov (United States)

    Pontes, Mauricio H; Lee, Eun-Jin; Choi, Jeongjoon; Groisman, Eduardo A

    2015-04-21

    Cellulose is the most abundant organic polymer on Earth. In bacteria, cellulose confers protection against environmental insults and is a constituent of biofilms typically formed on abiotic surfaces. We report that, surprisingly, Salmonella enterica serovar Typhimurium makes cellulose when inside macrophages. We determine that preventing cellulose synthesis increases virulence, whereas stimulation of cellulose synthesis inside macrophages decreases virulence. An attenuated mutant lacking the mgtC gene exhibited increased cellulose levels due to increased expression of the cellulose synthase gene bcsA and of cyclic diguanylate, the allosteric activator of the BcsA protein. Inactivation of bcsA restored wild-type virulence to the Salmonella mgtC mutant, but not to other attenuated mutants displaying a wild-type phenotype regarding cellulose. Our findings indicate that a virulence determinant can promote pathogenicity by repressing a pathogen's antivirulence trait. Moreover, they suggest that controlling antivirulence traits increases long-term pathogen fitness by mediating a trade-off between acute virulence and transmission.

  17. Salmonella promotes virulence by repressing cellulose production

    Science.gov (United States)

    Pontes, Mauricio H.; Lee, Eun-Jin; Choi, Jeongjoon; Groisman, Eduardo A.

    2015-01-01

    Cellulose is the most abundant organic polymer on Earth. In bacteria, cellulose confers protection against environmental insults and is a constituent of biofilms typically formed on abiotic surfaces. We report that, surprisingly, Salmonella enterica serovar Typhimurium makes cellulose when inside macrophages. We determine that preventing cellulose synthesis increases virulence, whereas stimulation of cellulose synthesis inside macrophages decreases virulence. An attenuated mutant lacking the mgtC gene exhibited increased cellulose levels due to increased expression of the cellulose synthase gene bcsA and of cyclic diguanylate, the allosteric activator of the BcsA protein. Inactivation of bcsA restored wild-type virulence to the Salmonella mgtC mutant, but not to other attenuated mutants displaying a wild-type phenotype regarding cellulose. Our findings indicate that a virulence determinant can promote pathogenicity by repressing a pathogen's antivirulence trait. Moreover, they suggest that controlling antivirulence traits increases long-term pathogen fitness by mediating a trade-off between acute virulence and transmission. PMID:25848006

  18. Soybean aphid (Hemiptera: Aphididae) response to soybean plant defense: stress levels, tradeoffs, and cross-virulence.

    Science.gov (United States)

    Enders, Laramy; Bickel, Ryan; Brisson, Jennifer; Heng-Moss, Tiffany; Siegfried, Blair; Zera, Anthony; Miller, Nick

    2014-02-01

    A variety of management methods to control the soybean aphid (Aphis glycines Matsumura) have been investigated since its invasion into North America in 2000, among them plant resistance has emerged as a viable option for reducing aphid damage to soybeans and preventing outbreaks. Plant resistance methods often use natural soybean plant defenses that impose stress on aphids by reducing fitness and altering behavior. Research efforts have heavily focused on identification and development of aphid resistant soybean varieties, leaving much unknown about soybean aphid response to stressful host plant defenses. In this study, we aimed to 1) evaluate lifetime fitness consequences and phenotypic variation in response to host plant-induced stress and 2) investigate whether trade-offs involving fitness costs and/or cross-virulence to multiple antibiotic soybean varieties exists. We compared aphid survival and reproduction during and after a short period of exposure to soybeans with the Rag2 resistance gene and measured aphid clonal variation in response to Rag2 soybeans. In addition, we measured the performance of Rag2 virulent and avirulent aphids on five soybean varieties with various forms of antibiotic resistance. Our results indicate that plant defenses impose high levels of stress and have long-term fitness consequences, even after aphids are removed from resistant plants. We identified one aphid clone that was able to colonize Rag2 among the seven clones tested, suggesting that virulent genotypes may be prevalent in natural populations. Finally, although we did not find evidence of cross-virulence to multiple antibiotic soybean varieties, our results suggest independent mechanisms of aphid virulence to Rag1 and Rag2 that may involve fitness costs.

  19. Virulence factors of the Mycobacterium tuberculosis complex

    Science.gov (United States)

    Forrellad, Marina A.; Klepp, Laura I.; Gioffré, Andrea; Sabio y García, Julia; Morbidoni, Hector R.; Santangelo, María de la Paz; Cataldi, Angel A.; Bigi, Fabiana

    2013-01-01

    The Mycobacterium tuberculosis complex (MTBC) consists of closely related species that cause tuberculosis in both humans and animals. This illness, still today, remains to be one of the leading causes of morbidity and mortality throughout the world. The mycobacteria enter the host by air, and, once in the lungs, are phagocytated by macrophages. This may lead to the rapid elimination of the bacillus or to the triggering of an active tuberculosis infection. A large number of different virulence factors have evolved in MTBC members as a response to the host immune reaction. The aim of this review is to describe the bacterial genes/proteins that are essential for the virulence of MTBC species, and that have been demonstrated in an in vivo model of infection. Knowledge of MTBC virulence factors is essential for the development of new vaccines and drugs to help manage the disease toward an increasingly more tuberculosis-free world. PMID:23076359

  20. Reproductive isolation during domestication.

    Science.gov (United States)

    Dempewolf, Hannes; Hodgins, Kathryn A; Rummell, Sonja E; Ellstrand, Norman C; Rieseberg, Loren H

    2012-07-01

    It has been hypothesized that reproductive isolation should facilitate evolution under domestication. However, a systematic comparison of reproductive barrier strength between crops and their progenitors has not been conducted to test this hypothesis. Here, we present a systematic survey of reproductive barriers between 32 economically important crop species and their progenitors to better understand the role of reproductive isolation during the domestication process. We took a conservative approach, avoiding those types of reproductive isolation that are poorly known for these taxa (e.g., differences in flowering time). We show that the majority of crops surveyed are isolated from their progenitors by one or more reproductive barriers, despite the fact that the most important reproductive barrier in natural systems, geographical isolation, was absent, at least in the initial stages of domestication for most species. Thus, barriers to reproduction between crops and wild relatives are closely associated with domestication and may facilitate it, thereby raising the question whether reproductive isolation could be viewed as a long-overlooked "domestication trait." Some of the reproductive barriers observed (e.g., polyploidy and uniparental reproduction), however, may have been favored for reasons other than, or in addition to, their effects on gene flow.

  1. Glucose starvation boosts Entamoeba histolytica virulence.

    Directory of Open Access Journals (Sweden)

    Ayala Tovy

    2011-08-01

    Full Text Available The unicellular parasite, Entamoeba histolytica, is exposed to numerous adverse conditions, such as nutrient deprivation, during its life cycle stages in the human host. In the present study, we examined whether the parasite virulence could be influenced by glucose starvation (GS. The migratory behaviour of the parasite and its capability to kill mammalian cells and to lyse erythrocytes is strongly enhanced following GS. In order to gain insights into the mechanism underlying the GS boosting effects on virulence, we analyzed differences in protein expression levels in control and glucose-starved trophozoites, by quantitative proteomic analysis. We observed that upstream regulatory element 3-binding protein (URE3-BP, a transcription factor that modulates E.histolytica virulence, and the lysine-rich protein 1 (KRiP1 which is induced during liver abscess development, are upregulated by GS. We also analyzed E. histolytica membrane fractions and noticed that the Gal/GalNAc lectin light subunit LgL1 is up-regulated by GS. Surprisingly, amoebapore A (Ap-A and cysteine proteinase A5 (CP-A5, two important E. histolytica virulence factors, were strongly down-regulated by GS. While the boosting effect of GS on E. histolytica virulence was conserved in strains silenced for Ap-A and CP-A5, it was lost in LgL1 and in KRiP1 down-regulated strains. These data emphasize the unexpected role of GS in the modulation of E.histolytica virulence and the involvement of KRiP1 and Lgl1 in this phenomenon.

  2. Gastrin-releasing peptide in the porcine pancreas

    DEFF Research Database (Denmark)

    Holst, J J; Poulsen, Steen Seier

    1987-01-01

    to consist of one main form, namely the 27-amino acid peptide originally extracted from porcine stomach, and small amounts of a C-terminal fragment identical with the C-terminal 10-amino acid peptide. Gastrin-releasing peptide-like immunoreactivity released from the isolated perfused porcine pancreas during...

  3. Growth of cultured porcine retinal pigment epithelial cells

    DEFF Research Database (Denmark)

    Wiencke, A.K.; Kiilgaard, Jens Folke; Nicolini, Jair

    2003-01-01

    To establish and characterize cultures of porcine retinal pigment epithelial (pRPE) cells in order to produce confluent monolayers of cells for transplantation.......To establish and characterize cultures of porcine retinal pigment epithelial (pRPE) cells in order to produce confluent monolayers of cells for transplantation....

  4. Comparison of gene expression patterns between porcine cumulus ...

    African Journals Online (AJOL)

    UPuser

    1 Department of Gene and Cell Engineering, Institute of Animal Science, Chinese Academy of Agricultural Sciences,. Beijing 100094 ... quantitative RT-PCR methods, we compared the mRNA expression patterns in porcine oocytes from two ... Keywords: Differential gene expression, DD-RT-PCR, porcine oocytes, cumulus.

  5. Factors influencing transmission of porcine cysticercosis in Tanzania

    DEFF Research Database (Denmark)

    Braae, Uffe Christian; Wendy, Harrison; Magnussen, Pascal

    porcine cysticercosis could be associated with absence or completely open latrines (p=0.035, OR 5.98, CI: 1.33- 43.02) compared to enclosed latrines, and feeding potato peels to pigs (P=0.007, OR 3.45, CI: 1.43-8.79). Prevalence of porcine cysticercosis fluctuated throughout the seasons, and confined pigs...

  6. Determination of free amino acids of porcine serum responsible for ...

    African Journals Online (AJOL)

    Determination of free amino acids of porcine serum responsible for the meat quality by 1 H NMR and HPLC analyses. ... African Journal of Biotechnology ... The pH24h correlated well with the water holding capacity (WHC) of porcine meat, whereas a strongly negative correlation was observed between pH24h and serum ...

  7. Comparative histology and immunohistochemistry of porcine versus human skin.

    Science.gov (United States)

    Debeer, Sabine; Le Luduec, Jean-Benoît; Kaiserlian, Dominique; Laurent, Philippe; Nicolas, Jean-François; Dubois, Bertrand; Kanitakis, Jean

    2013-01-01

    Porcine skin is increasingly being employed as a model of human skin in various research fields, including pharmacology, toxicology and immunology, with particular interest in percutaneous permeation and organ transplantation. Porcine skin shows several anatomical and physiological similarities, but also some differences, with human skin, but few in depth comparative studies are so far available. To study the immunohistochemical properties of normal porcine skin in comparison with human skin. We performed a histological and immunohistochemical study on frozen and formalin-fixed, paraffin-embedded skin biopsies from domestic swine and normal human skin, using a panel of 93 monoclonal or polyclonal antibodies recognizing various human and porcine skin cell types or structures. We found that several antibodies used to detect normal human skin cells showed equivalent immunoreactivity on normal porcine skin. However, some antibodies commonly used to detect human skin antigens remained unreactive on porcine skin. Our findings highlight the main immunohistochemical properties of porcine skin in comparison with those of human skin and provide a morphological and immunohistochemical basis useful to researchers using porcine skin.

  8. Reproduction (II): Human Control of Reproductive Processes

    Science.gov (United States)

    Jost, Alfred

    1970-01-01

    Describes methods of intervening in reproduction of animals and humans (artificial insemination, contraception, ovular and blastodisc transplants, pre selection of sex, cloning) and discusses the social implications of their use with humans. (AL)

  9. BMP15 Prevents Cumulus Cell Apoptosis Through CCL2 and FBN1 in Porcine Ovaries

    Directory of Open Access Journals (Sweden)

    Bo Zhai

    2013-07-01

    Full Text Available Background: Bone morphogenetic protein-15 (BMP15 is a maternal gene necessary for mammalian reproduction. BMP15 expression increased in oocytes accompanied by follicle growth and development. The function and regulation mechanism of BMP15 in porcine cumulus cell apoptosis process is still unclear now. Methods: In this study, flow cytometry (FCM was used to analyze the effects of BMP15 with different concentrations to cumulus cell apoptosis. High-throughput sequencing technology was carried out to screen regulatory genes linked closely with BMP15. In order to confirm the function of (MCP-1/CCL2 and FBN1 in cumulus cell apoptosis, RNA interference (RNAi method was used to inhibit the expression of (MCP-1/CCL2 and FBN1. Apoptosis and proliferation of cumulus cell treated with siRNA transfection technology were measured by FCM, 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide, quantitative real time-PCR (RT-qPCR and western blotting. Results: The results showed that the apoptosis levels of cumulus cell treated by BMP15 decreased significantly in a dose-dependent manner. The expression of related genes protein 1 (MCP-1/CCL2 and fibrillin1 (FBN1 were both regulated by BMP15. After transfection, the proliferation of porcine cumulus cells increased significantly and apoptosis of cumulus cells was prevented while FBN1 was silenced after BMP15 treatment. The proliferation of cumulus cells decreased significantly and apoptosis rate of cumulus cells increased significantly while CCL2 was silenced. Conclusion: The results obtained in this study firstly demonstrated that CCL2 and FBN1 are important regulatory factors of BMP15 in preventing cumulus cell apoptosis in porcine ovaries.

  10. Mycotoxin alpha-zearalenol impairs the quality of preimplantation porcine embryos.

    Science.gov (United States)

    Wang, Hongfeng; Camargo Rodriguez, Omar; Memili, Erdogan

    2012-01-01

    Alpha-Zearalenol (α-ZEA) is one of derivatives from Zearalenone (ZEA) which impacts mammalian reproduction and development. Previous studies have shown that pigs are sensitive to the estradiol-like effects of α-ZEA. However, the effect of α-ZEA for the early embryonic development has not been fully studied. The objective of this study was to identify the direct toxicity of α-ZEA on porcine preimplantation embryonic development, embryo quality and expression of developmentally important genes. Presumptive zygotes were cultured in porcine zygote medium 3 (PZM-3) in the presence of α-ZEA (n=2,957) or 17β-estradiol (E2) (n=1,333) dissolved in 0.1% Dimethyl Sulfoxide (DMSO) from 24 to 84 h post insemination followed by determination of apoptotic cell numbers and transcript levels of BAX, BCL2L1 and POU5F1 in blastocysts. Cleavage rates on day 2 were significantly decreased in 10, 30 and 60 µM α-ZEA groups; whereas blastocyst rates on day 6 were significantly decreased in the 30 and 60 µM of α-ZEA groups. Only the 100 µM E2 group significantly decreased cleavage and blastocyst rates. Total cell numbers (TCN) in blastocysts were significantly lower in the 10 µM α-ZEA group, but no differences in apoptotic cell rates were found. The expression levels of POU5F1 and BCL2L1 transcripts were similar; however, levels of BAX transcripts and the BAX/BCL2L1 ratio were increased in both α-ZEA groups. Since α-ZEA and E2 did not elicit similar effects, results suggest that α-ZEA might impact porcine preimplantation embryonic development through pathways other than estrogen receptor binding.

  11. Analysis of the Binding Sites of Porcine Sialoadhesin Receptor with PRRSV

    Directory of Open Access Journals (Sweden)

    Yibo Jiang

    2013-12-01

    Full Text Available Porcine reproductive and respiratory syndrome virus (PRRSV can infect pigs and cause enormous economic losses to the pig industry worldwide. Porcine sialoadhesin (pSN and CD163 have been identified as key viral receptors on porcine alveolar macrophages (PAM, a main target cell infected by PRRSV. In this study, the protein structures of amino acids 1–119 from the pSN and cSN (cattle sialoadhesin N-termini (excluding the 19-amino acid signal peptide were modeled via homology modeling based on mSN (mouse sialoadhesin template structures using bioinformatics tools. Subsequently, pSN and cSN homology structures were superposed onto the mSN protein structure to predict the binding sites of pSN. As a validation experiment, the SN N-terminus (including the wild-type and site-directed-mutant-types of pSN and cSN was cloned and expressed as a SN-GFP chimera protein. The binding activity between SN and PRRSV was confirmed by WB (Western blotting, FAR-WB (far Western blotting, ELISA (enzyme-linked immunosorbent assay and immunofluorescence assay. We found that the S107 amino acid residue in the pSN N-terminal played a crucial role in forming a special cavity, as well as a hydrogen bond for enhancing PRRSV binding during PRRSV infection. S107 may be glycosylated during PRRSV infection and may also be involved in forming the cavity for binding PRRSV along with other sites, including W2, Y44, S45, R97, R105, W106 and V109. Additionally, S107 might also be important for pSN binding with PRRSV. However, the function of these binding sites must be confirmed by further studies.

  12. Progress, problems and prospects of porcine pluripotent stem cells

    Directory of Open Access Journals (Sweden)

    Hanning WANG,Yangli PEI,Ning LI,Jianyong HAN

    2014-02-01

    Full Text Available Pluripotent stem cells (PSCs, including embryonic stem cells (ESCs and induced PSCs (iPSCs, can differentiate into cells of the three germ layers, suggesting that PSCs have great potential for basic developmental biology research and wide applications for clinical medicine. Genuine ESCs and iPSCs have been derived from mice and rats, but not from livestock such as the pig─an ideal animal model for studying human disease and regenerative medicine due to similarities with human physiologic processes. Efforts to derive porcine ESCs and iPSCs have not yielded high-quality PSCs that can produce chimeras with germline transmission. Thus, exploration of the unique porcine gene regulation network of preimplantation embryonic development may permit optimization of in vitro culture systems for raising porcine PSCs. Here we summarize the recent progress in porcine PSC generation as well as the problems encountered during this progress and we depict prospects for generating porcine naive PSCs.

  13. Splice variants of porcine PPHLN1 encoding periphilin-1

    DEFF Research Database (Denmark)

    Larsen, Knud Erik; Momeni, Jamal; Farajzadeh, Leila

    2017-01-01

    of the periphilin-1 protein. Thus, variants Sp1 and Sp1 are the result of alternative splicing. The porcine PPHLN1 gene was mapped to chromosome 5. The porcine PPHLN1 gene was found to be differentially expressed in various porcine organs and tissues. The sequence of the porcine PPHLN1 cDNA, encoding the periphilin......The periphilin-1 protein is encoded by the PPHLN1 gene. Periphilin-1 is found in the cornified cell envelope during the terminal differentiation of keratinocyte at the outer layer of epidermis. In the current study we report on the cloning and characterization of the porcine PPHLN1 cDNA and two...... splice variants hereof. RT-PCR cloning using oligonucleotide primers derived from in silico sequences resulted in three PPHLN1 transcripts: a full-length mRNA and two transcript variant resulting in shorter proteins. The longest encoded periphilin-1, consisting of 373 amino acids, displays a high...

  14. Synthesis of biologically active porcine secretin and [ITyr10] porcine secretin

    DEFF Research Database (Denmark)

    Kofod, Hans

    1991-01-01

    Porcine secretin, [Tyr10] secretin, and [Tyr13] secretin were synthesized by solid phase methodology and purified by stepwise gradient elution from a short reversed-phase column with ethanol and acetic acid as organic modifiers. [Tyr10] secretin and [Tyr13] secretin were iodinated by the chloramine-T...

  15. Rapid and specific detection of porcine parvovirus using real-time PCR and high resolution melting (HRM) analysis.

    Science.gov (United States)

    Yu, Hai-Qiong; Cai, Xian-Quan; Lin, Zhi-Xiong; Li, Xiang-Li; Yue, Qiao-Yun; Li, Rong; Zhu, Xing-Quan

    2015-02-28

    Porcine parvovirus (PPV) is the important causative agent for infectious infertility, which is a fairly tough virus that multiplies normally in the intestine of pigs without causing clinical signs in the world. We developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the detection of PPV. Primers targeting the VP gene were highly specific, as evidenced by the negative amplification of closely related viruses, such as porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The performance of unlabeled real time PCR was compared to TaqMan real time PCR, and the detection limits of the two methods were nearly equal. Moreover, there was good correlation between Cp and diluted genomic DNA when tested with the two methods. The assay has the accuracy of 100% in reference to labeled real time PCR, when it was tested on 45 clinical samples. The present study demonstrated that the established assay integrating real-time PCR and HRM is relatively cost-effective and more stable, which provides an alternative tool for rapid, simple, specific and sensitive detection of PPV.

  16. A magnetic nanoparticle-based multiple-gene delivery system for transfection of porcine kidney cells.

    Directory of Open Access Journals (Sweden)

    Yan Wang

    Full Text Available Superparamagnetic nanoparticles are promising candidates for gene delivery into mammalian somatic cells and may be useful for reproductive cloning using the somatic cell nuclear transfer technique. However, limited investigations of their potential applications in animal genetics and breeding, particularly multiple-gene delivery by magnetofection, have been performed. Here, we developed a stable, targetable and convenient system for delivering multiple genes into the nuclei of porcine somatic cells using magnetic Fe3O4 nanoparticles as gene carriers. After surface modification by polyethylenimine, the spherical magnetic Fe3O4 nanoparticles showed strong binding affinity for DNA plasmids expressing the genes encoding a green (DNAGFP or red (DNADsRed fluorescent protein. At weight ratios of DNAGFP or DNADsRed to magnetic nanoparticles lower than or equal to 10∶1 or 5∶1, respectively, the DNA molecules were completely bound by the magnetic nanoparticles. Atomic force microscopy analyses confirmed binding of the spherical magnetic nanoparticles to stretched DNA strands up to several hundred nanometers in length. As a result, stable and efficient co-expression of GFP and DsRed in porcine kidney PK-15 cells was achieved by magnetofection. The results presented here demonstrate the potential application of magnetic nanoparticles as an attractive delivery system for animal genetics and breeding studies.

  17. Pathotyping and antibiotic resistance of porcine enterovirulent Escherichia coli strains from Switzerland (2014-2015).

    Science.gov (United States)

    Brand, P; Gobeli, S; Perreten, V

    2017-07-01

    A total of 131 porcine E. coli were isolated in 2014 and 2015 from the gut of 115 pigs raised in Switzerland and suffering from diarrhea. The isolates were tested for antibiotic resistance, serotypes, virulence factors and genetic diversity. Serotypes were assigned by agglutination tests and virulence genes were identified by polymerase chain reaction (PCR). Antibiotic resistance profile was determined by the measurement of the MIC of 14 antibiotics and by the detection of the corresponding genes using microarray and PCR approaches. Genetic diversity was determined by repetitive palindromic PCR (rep- PCR) revealing a heterogenous population. Half of the E. coli isolates possessing virulence factors could not be assigned to any of the 19 serotypes tested, but contained toxins and adhesins similarly to the sero-typable E. coli isolates. The most prevalent E. coli serotypes found were K88ac (18%), O139:K82 (6%), O141:K85ac (5%), O108:K`V189` (5%), O119:K`V113` (3%) and O157:K`V17` (2%). The combination of toxins EAST-1, STb and LT-I and adhesin F4 characterizing ETEC was the most frequent. The shigatoxin Stx2e (STEC) and intimin Eae (EPEC) were also detected, but less frequently. Seventy percent of the isolates were resistant to at least one antibiotic and 29% were resistant to more than 3 antibiotics. Isolates exhibited resistance to tetracycline (50%) associated to resistance genes tet(A), tet(B) and tet(C), sulfamethoxazole (49%) [sul1, sul2 and sul3], trimethoprim (34%) [dfr], nalidixic acid (29%), ampicillin (26%) [blaTEM-1], gentamicin (17%) [aac(3) -IIc, aac(3) -IVa and aac(3) -VIa], chloramphenicol (17%) [catAI and catAIII], and ciprofloxacin (8%) [mutations in GyrA (S83L) and ParC (S80I)]. All isolates were susceptible to 3rd generation cephalosporins, carbapenems, colistin and tigecycline. Pathogenic E. coli isolates from pigs in Switzerland could frequently not be assigned to a known serotype even if they contained diarrhea-causing virulence factors. They

  18. Rare Helicobacter pylori Virulence Genotypes in Bhutan.

    Science.gov (United States)

    Matsunari, Osamu; Miftahussurur, Muhammad; Shiota, Seiji; Suzuki, Rumiko; Vilaichone, Ratha-Korn; Uchida, Tomohisa; Ratanachu-ek, Thawee; Tshering, Lotay; Mahachai, Varocha; Yamaoka, Yoshio

    2016-03-02

    Both the prevalence of Helicobacter pylori infection and the incidence of gastric cancer are high in Bhutan. The high incidence of atrophic gastritis and gastric cancer suggest the phylogeographic origin of an infection with a more virulent strain of H. pylori. More than 90% of Bhutanese strains possessed the highly virulent East Asian-type CagA and all strains had the most virulent type of vacA (s1 type). More than half also had multiple repeats in East Asian-type CagA, which are rare in other countries and are reported characteristictly found in assciation with atrophic gastritis and gastric cancer consistent with Bhutanese strains having multiple H. pylori virulence factors associated with an increase in gastric cancer risk. Phylogeographic analyses showed that most Bhutanese strains belonged to the East Asian population type with some strains (17.5%) sharing East Asian and Amerindian components. Only 9.5% belonged to the European type consistant with H. pylori in Bhutan representing an intermediate evolutionary stage between H. pylori from European and East Asian countries.

  19. NEW VIRULENCE FACTORS OF STREPTOCOCCUS PNEUMONIAE

    NARCIS (Netherlands)

    Hermans, Peter Wilhelmus Maria; Bootsma, Jeanette Hester; Burghout, Pieter Jan; Kuipers, Oscar; Bijlsma, Johanna Jacoba Elisabeth; Kloosterman, Tomas Gerrit; Andersen, Christian O.

    2011-01-01

    The present invention provides proteins/genes, which are essential for survival, and consequently, for virulence of Streptococcus pneumoniae in vivo, and thus are ideal vaccine candidates for a vaccine preparation against pneumococcal infection. Further, also antibodies against said protein(s) are

  20. Salmonella virulence plasmid: pathogenesis and ecology.

    Science.gov (United States)

    Silva, Claudia; Puente, José Luis; Calva, Edmundo

    2017-06-22

    A current view on the role of the Salmonella virulence plasmid in the pathogenesis of animal and human hosts is discussed; including the possible relevance in secondary ecological niches. Various strategies towards further studies in this respect are proposed within the One Health Concept. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Efflux inhibitor suppresses Streptococcus mutans virulence properties.

    Science.gov (United States)

    Zeng, Huihui; Liu, Jia; Ling, Junqi

    2017-04-01

    It is well established that efflux pumps play important roles in bacterial pathogenicity and efflux inhibitors (EIs) have been proved to be effective in suppressing bacterial virulence properties. However, little is known regarding the EI of Streptococcus mutans, a well-known caries-inducing bacterium. In this study, we identified the EI of S. mutans through ethidium bromide efflux assay and investigated how EI affected S. mutans virulence regarding the cariogenicity and stress response. Results indicated that reserpine, the identified EI, suppressed acid tolerance, mutacin production and transformation efficiency of S. mutans, and modified biofilm architecture and extracellular polysaccharide distribution. Suppressed glycosyltransferase activity was also noted after reserpine exposure. The data from quantitative real-time-PCR demonstrated that reserpine significantly altered the expression profile of quorum-sensing and virulence-associated genes. These findings suggest that reserpine represents a promising adjunct anticariogenic agent in that it suppresses virulence properties of S. mutans. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Porcine lung surfactant protein B gene (SFTPB)

    DEFF Research Database (Denmark)

    Cirera Salicio, Susanna; Fredholm, Merete

    2008-01-01

    The porcine surfactant protein B (SFTPB) is a single copy gene on chromosome 3. Three different cDNAs for the SFTPB have been isolated and sequenced. Nucleotide sequence comparison revealed six nonsynonymous single nucleotide polymorphisms (SNPs), four synonymous SNPs and an in-frame deletion of 69...... bp in the region coding for the active protein. Northern analysis showed lung-specific expression of three different isoforms of the SFTPB transcript. The expression level for the SFTPB gene is low in 50 days-old fetus and it increases during lung development. Quantitative real-time polymerase chain...

  3. Survey on porcine trichinellosis in Ecuador

    DEFF Research Database (Denmark)

    Chávez-Larrea, M. A.; Dorny, P.; Møller, L. N.

    2004-01-01

    A survey on porcine trichinellosis was organised in Ecuador between 2000 and 2003. Blood samples were taken in slaughterhouses (study 1, n = 2000; study 2, n = 331) and in a remote village where pigs are free roaming (study 3, n = 646) and examined by ELISA using excretory/secretory (E/S) antigens...... that Trichinella is present in Ecuador; however, prevalence and parasite burdens are likely to be very low. The likelihood of detecting trichinellosis are higher in traditional settings than in pigs raised on improved farms...

  4. Survey of porcine parvovirus infection in swine fetuses and their dams at a Minnesota abattoir

    Energy Technology Data Exchange (ETDEWEB)

    Thacker, B.J.; Leman, A.D.; Hurtgen, J.P.; Sauber, T.E.; Joo, H.S.

    1981-05-01

    Reproductive tracts were recovered from 209 sow and 32 gilt carcasses at slaughter; animals had been pregnant not less than 27 days. Of 241 litters examined, 28 (11.6%) contained one or more porcine parvovirus (PPV)-infected fetuses, as determined by immunofluorescent microscopy. The frequencies in sow and gilt litters were 12.0% and 9.4%, respectively. The PPV antigen was detected in 219 of 334 (65.6%) dead or mummified fetuses and in 12 of 2,172 (0.5%) live fetuses examined. The 18 litters which contained only dead or mummified fetuses were infected with PPV. As the percentage of litter mummification increased, the likelihood of finding PPV increased. The PPV antibody was detected in ovarian follicular fluids of 94.3% of the sows and 78.1% of the gilts. These findings indicate that PPV is highly associated with fetal mummification and that some pregnant gilts and sows are susceptible to infection.

  5. Systemic Approach to Virulence Gene Network Analysis for Gaining New Insight into Cryptococcal Virulence

    Directory of Open Access Journals (Sweden)

    Antoni N Malachowski

    2016-10-01

    Full Text Available Cryptococcus neoformans is pathogenic yeast, responsible for highly lethal infections in compromised patients around the globe. C. neoformans typically initiates infections in mammalian lung tissue and subsequently disseminates to the central nervous system where it causes significant pathologies. Virulence genes of C. neoformans are being characterized at an increasing rate, however, we are far from a comprehensive understanding of their roles and genetic interactions. Some of these reported virulence genes are scattered throughout different databases, while others are not yet included. This study gathered and analyzed 150 reported virulence associated factors (VAFs of C. neoformans. Using the web resource STRING database, our study identified different interactions between the total VAFs and those involved specifically in lung and brain infections and identified a new strain specific virulence gene, sho1, involved in the mitogen-activated protein kinase signaling pathway. As predicted by our analysis, sho1 expression enhanced C. neoformans virulence in a mouse model of pulmonary infection, contributing to enhanced non-protective immune Th2 bias and progressively enhancing fungal growth in the infected lungs. Sequence analysis indicated 77.4% (116 of total studied VAFs are soluble proteins, and 22.7% (34 are transmembrane proteins. Motifs involved in regulation and signaling such as protein kinases and transcription factors are highly enriched in Cryptococcus VAFs. Altogether, this study represents a pioneering effort in analysis of the virulence composite network of C. neoformans using a systems biology approach.

  6. Sample collection of virulent and non-virulent B. anthracis and Y. pestis for bioforensics analysis

    Energy Technology Data Exchange (ETDEWEB)

    Hong-geller, Elizabeth [Los Alamos National Laboratory; Valdez, Yolanda E [Los Alamos National Laboratory; Shou, Yulin [Los Alamos National Laboratory; Yoshida, Thomas M [Los Alamos National Laboratory; Marrone, Babetta L [Los Alamos National Laboratory; Dunbar, John [Los Alamos National Laboratory

    2009-01-01

    Validated sample collection methods are needed for recovery of microbial evidence in the event of accidental or intentional release of biological agents into the environment. To address this need, we evaluated the sample recovery efficiencies of two collection methods -- swabs and wipes -- for both non-virulent and virulent strains of B. anthracis and Y. pestis from four types of non-porous surfaces: two hydrophilic surfaces, stainless steel and glass, and two hydrophobic surfaces, vinyl and plastic. Sample recovery was quantified using Real-time qPCR to assay for intact DNA signatures. We found no consistent difference in collection efficiency between swabs or wipes. Furthermore, collection efficiency was more surface-dependent for virulent strains than non-virulent strains. For the two non-virulent strains, B. anthracis Sterne and Y. pestis A1122, collection efficiency was approximately 100% and 1 %, respectively, from all four surfaces. In contrast, recovery of B. anthracis Ames spores and Y. pestis C092 from vinyl and plastic was generally lower compared to collection from glass or stainless steel, suggesting that surface hydrophobicity may playa role in the strength of pathogen adhesion. The surface-dependent collection efficiencies observed with the virulent strains may arise from strain-specific expression of capsular material or other cell surface receptors that alter cell adhesion to specific surfaces. These findings contribute to validation of standard bioforensics procedures and emphasize the importance of specific strain and surface interactions in pathogen detection.

  7. Systemic Approach to Virulence Gene Network Analysis for Gaining New Insight into Cryptococcal Virulence.

    Science.gov (United States)

    Malachowski, Antoni N; Yosri, Mohamed; Park, Goun; Bahn, Yong-Sun; He, Yongqun; Olszewski, Michal A

    2016-01-01

    Cryptococcus neoformans is pathogenic yeast, responsible for highly lethal infections in compromised patients around the globe. C. neoformans typically initiates infections in mammalian lung tissue and subsequently disseminates to the central nervous system where it causes significant pathologies. Virulence genes of C. neoformans are being characterized at an increasing rate, however, we are far from a comprehensive understanding of their roles and genetic interactions. Some of these reported virulence genes are scattered throughout different databases, while others are not yet included. This study gathered and analyzed 150 reported virulence associated factors (VAFs) of C. neoformans. Using the web resource STRING database, our study identified different interactions between the total VAFs and those involved specifically in lung and brain infections and identified a new strain specific virulence gene, SHO1, involved in the mitogen-activated protein kinase signaling pathway. As predicted by our analysis, SHO1 expression enhanced C. neoformans virulence in a mouse model of pulmonary infection, contributing to enhanced non-protective immune Th2 bias and progressively enhancing fungal growth in the infected lungs. Sequence analysis indicated 77.4% (116) of total studied VAFs are soluble proteins, and 22.7% (34) are transmembrane proteins. Motifs involved in regulation and signaling such as protein kinases and transcription factors are highly enriched in Cryptococcus VAFs. Altogether, this study represents a pioneering effort in analysis of the virulence composite network of C. neoformans using a systems biology approach.

  8. Reproductive Disorders in Parrots.

    Science.gov (United States)

    Scagnelli, Alyssa M; Tully, Thomas N

    2017-05-01

    Disease affecting the reproductive tract of the companion parrot is often impacted by physiologic and environmental stimuli. In conjunction with appropriate medical management, some birds diagnosed with reproductive disorders may be successfully treated. Once the bird is diagnosed with a disease condition affecting the reproductive tract, therapeutic measures are focused on stabilizing and supporting the patient, and surgical intervention is required only in the most severe cases. Hormonal therapy with synthetic, long-acting GnRH agonists should be considered for chronic reproductive disease conditions in which decreasing ovarian activity can help alleviate certain disease processes. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Reproductive and developmental toxicology

    National Research Council Canada - National Science Library

    Gupta, Ramesh C

    2011-01-01

    .... Reproductive and Developmental Toxicology is a comprehensive and authoritative resource providing the latest literature enriched with relevant references describing every aspect of this area of science...

  10. Short Rotations in Forest Plantations Accelerate Virulence Evolution in Root-Rot Pathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Jean-Paul Soularue

    2017-06-01

    Full Text Available As disease outbreaks in forest plantations are causing concern worldwide, a clear understanding of the influence of silvicultural practices on the development of epidemics is still lacking. Importantly, silvicultural practices are likely to simultaneously affect epidemiological and evolutionary dynamics of pathogen populations. We propose a genetically explicit and individual-based model of virulence evolution in a root-rot pathogenic fungus spreading across forest landscapes, taking the Armillaria ostoyae–Pinus pinaster pathosystem as reference. We used the model to study the effects of rotation length on the evolution of virulence and the propagation of the fungus within a forest landscape composed of even-aged stands regularly altered by clear-cutting and thinning operations. The life cycle of the fungus modeled combines asexual and sexual reproduction modes, and also includes parasitic and saprotrophic phases. Moreover, the tree susceptibility to the pathogen is primarily determined by the age of the stand. Our simulations indicated that the shortest rotation length accelerated both the evolution of virulence and the development of the epidemics, whatever the genetic variability in the initial fungal population and the asexuality rate of the fungal species

  11. Microarray analysis of the effect of Streptococcus equi subsp. zooepidemicus M-like protein in infecting porcine pulmonary alveolar macrophage.

    Directory of Open Access Journals (Sweden)

    Zhe Ma

    Full Text Available Streptococcus equi subsp. zooepidemicus (S. zooepidemicus, which belongs to Lancefield group C streptococci, is an important pathogen of domesticated species, causing septicemia, meningitis and mammitis. M-like protein (SzP is an important virulence factor of S. zooepidemicus and contributes to bacterial infection and antiphagocytosis. To increase our knowledge of the mechanism of SzP in infection, we profiled the response of porcine pulmonary alveolar macrophage (PAM to infection with S. zooepidemicus ATCC35246 wild strain (WD and SzP-knockout strain (KO using the Roche NimbleGen Porcine Genome Expression Array. We found SzP contributed to differential expression of 446 genes, with upregulation of 134 genes and downregulation of 312 genes. Gene Ontology category and KEGG pathway were analyzed for relationships among differentially expressed genes. These genes were represented in a variety of functional categories, including genes involved in immune response, regulation of chemokine production, signal transduction and regulation of apoptosis. The reliability of the data obtained from the microarray was verified by performing quantitative real-time PCR on 12 representative genes. The data will contribute to understanding of SzP mediated mechanisms of S. zooepidemicus pathogenesis.

  12. Growth and survival of the fish pathogenic bacterium, Flavobacterium columnare, in tilapia mucus and porcine gastric mucin.

    Science.gov (United States)

    Shoemaker, Craig A; LaFrentz, Benjamin R

    2015-02-01

    Flavobacterium columnare, an economically important Gram-negative bacterium of freshwater farmed fish, colonizes the skin and gills in the initial steps of pathogenesis. The surface of fish is coated with mucus made up of high molecular weight glycoproteins. Limited studies have described the ability of bacterial pathogens to grow in fish mucus. Our objective was to determine if F. columnare isolates could grow and survive in formulated water (FW) containing autoclaved tilapia mucus or porcine gastric mucin. We demonstrated the ability of F. columnare genomovars I, II, II-B and III to replicate (2-3 logs) and survive (21 to >100 days) in FW containing tilapia mucus. In a second experiment, genomovar I and II isolates were found to replicate in FW containing tilapia mucus or porcine mucin but not in FW only. From a practical standpoint, fish handling and/or hauling results in stress that leads to mucus sloughing often with subsequent F. columnare infection. Flavobacterium columnare utilizes fish mucus as a nutrient source, and studies are underway to determine if growth in mucus or mucin results in differential protein expression and/or increased virulence of F. columnare towards fish. © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. [Porcine stress syndrome and PSE meat: clinical symptoms, pathogenesis, etiology and animal rights aspects].

    Science.gov (United States)

    Wendt, M; Bickhardt, K; Herzog, A; Fischer, A; Martens, H; Richter, T

    2000-05-01

    A review is given about the clinical symptoms, pathogenesis and aetiology of the porcine stress syndrome, furthermore aspects of animal welfare are discussed. The current breeding programmes of pig industry in Germany in many cases include animals with a mutation of the ryanodine-receptor (RYR-1)-gene--homozygous or heterozygous. This situation is the result of an intensive breeding of pigs during the last decades with the intention of increased lean carcass content and corresponding proceeds. The homozygous pigs are more stress susceptible (porcine stress syndrome) and produce meat of poor quality (PSE), which is also the case to some extend in heterozygous animals. The clinical symptoms of this muscle disease are characterised by a deficit of oxygen and a rapid glycolysis accompanied by a production of lactic acid and acidosis primarily in II B white muscle fibres. There is no doubt that a very close causal relation exists between the mutation of the RYR-1 and the porcine stress syndrome as well as the poor meat quality. The present knowledge of this disease, the genetic background, the physiology and pathophysiology of the mutation of the RYR-1 leads to the imperative conclusion to eliminate this mutated RYR-1 by selection of healthy pigs, which has been done successfully in other countries with important pig production. This conclusion is also supported by simple economic reasons because fertility, reproduction and daily weight gain are significantly reduced in stress susceptible pigs. Furthermore, it should be emphasised that regular breeding with the mutated RYR-1 is also a matter of animal welfare. The evident correlation between the mutated RYR-1 and the porcine stress syndrome, which includes degeneration of the muscle, pain and even life threatening malignant hyperthermia, can easily lead to the accusation in the public that diseased animals are used for pig meat production. Consequently, the authors would like to urge the breeding companies and the

  14. Telomere reprogramming and maintenance in porcine iPS cells.

    Directory of Open Access Journals (Sweden)

    Guangzhen Ji

    Full Text Available Telomere reprogramming and silencing of exogenous genes have been demonstrated in mouse and human induced pluripotent stem cells (iPS cells. Pigs have the potential to provide xenotransplant for humans, and to model and test human diseases. We investigated the telomere length and maintenance in porcine iPS cells generated and cultured under various conditions. Telomere lengths vary among different porcine iPS cell lines, some with telomere elongation and maintenance, and others telomere shortening. Porcine iPS cells with sufficient telomere length maintenance show the ability to differentiate in vivo by teratoma formation test. IPS cells with short or dysfunctional telomeres exhibit reduced ability to form teratomas. Moreover, insufficient telomerase and incomplete telomere reprogramming and/or maintenance link to sustained activation of exogenous genes in porcine iPS cells. In contrast, porcine iPS cells with reduced expression of exogenous genes or partial exogene silencing exhibit insufficient activation of endogenous pluripotent genes and telomerase genes, accompanied by telomere shortening with increasing passages. Moreover, telomere doublets, telomere sister chromatid exchanges and t-circles that presumably are involved in telomere lengthening by recombination also are found in porcine iPS cells. These data suggest that both telomerase-dependent and telomerase-independent mechanisms are involved in telomere reprogramming during induction and passages of porcine iPS cells, but these are insufficient, resulting in increased telomere damage and shortening, and chromosomal instability. Active exogenes might compensate for insufficient activation of endogenous genes and incomplete telomere reprogramming and maintenance of porcine iPS cells. Further understanding of telomere reprogramming and maintenance may help improve the quality of porcine iPS cells.

  15. Telomere reprogramming and maintenance in porcine iPS cells.

    Science.gov (United States)

    Ji, Guangzhen; Ruan, Weimin; Liu, Kai; Wang, Fang; Sakellariou, Despoina; Chen, Jijun; Yang, Yang; Okuka, Maja; Han, Jianyong; Liu, Zhonghua; Lai, Liangxue; Gagos, Sarantis; Xiao, Lei; Deng, Hongkui; Li, Ning; Liu, Lin

    2013-01-01

    Telomere reprogramming and silencing of exogenous genes have been demonstrated in mouse and human induced pluripotent stem cells (iPS cells). Pigs have the potential to provide xenotransplant for humans, and to model and test human diseases. We investigated the telomere length and maintenance in porcine iPS cells generated and cultured under various conditions. Telomere lengths vary among different porcine iPS cell lines, some with telomere elongation and maintenance, and others telomere shortening. Porcine iPS cells with sufficient telomere length maintenance show the ability to differentiate in vivo by teratoma formation test. IPS cells with short or dysfunctional telomeres exhibit reduced ability to form teratomas. Moreover, insufficient telomerase and incomplete telomere reprogramming and/or maintenance link to sustained activation of exogenous genes in porcine iPS cells. In contrast, porcine iPS cells with reduced expression of exogenous genes or partial exogene silencing exhibit insufficient activation of endogenous pluripotent genes and telomerase genes, accompanied by telomere shortening with increasing passages. Moreover, telomere doublets, telomere sister chromatid exchanges and t-circles that presumably are involved in telomere lengthening by recombination also are found in porcine iPS cells. These data suggest that both telomerase-dependent and telomerase-independent mechanisms are involved in telomere reprogramming during induction and passages of porcine iPS cells, but these are insufficient, resulting in increased telomere damage and shortening, and chromosomal instability. Active exogenes might compensate for insufficient activation of endogenous genes and incomplete telomere reprogramming and maintenance of porcine iPS cells. Further understanding of telomere reprogramming and maintenance may help improve the quality of porcine iPS cells.

  16. Melatonin regulates lipid metabolism in porcine oocytes.

    Science.gov (United States)

    Jin, Jun-Xue; Lee, Sanghoon; Taweechaipaisankul, Anukul; Kim, Geon A; Lee, Byeong Chun

    2017-03-01

    It is being increasingly recognized that the processes of lipogenesis and lipolysis are important for providing an essential energy source during oocyte maturation and embryo development. Recent studies demonstrated that melatonin has a role in lipid metabolism regulation, including lipogenesis, lipolysis, and mitochondrial biogenesis. In this study, we attempted to investigate the effects of melatonin on lipid metabolism during porcine oocyte in vitro maturation. Melatonin treatment significantly enhanced the number of lipid droplets (LDs) and upregulated gene expression related to lipogenesis (ACACA, FASN, PPARγ, and SREBF1). Oocytes treated with melatonin formed smaller LDs and abundantly expressed several genes associated with lipolysis, including ATGL, CGI-58, HSL, and PLIN2. Moreover, melatonin significantly increased the content of fatty acids, mitochondria, and ATP, as indicated by fluorescent staining. Concomitantly, melatonin treatment upregulated gene expression related to fatty acid β-oxidation (CPT1a, CPT1b, CPT2, and ACADS) and mitochondrial biogenesis (PGC-1α, TFAM, and PRDX2). Overall, melatonin treatment not only altered both the morphology and amount of LDs, but also increased the content of fatty acids, mitochondria, and ATP. In addition, melatonin upregulated mRNA expression levels of lipogenesis, lipolysis, β-oxidation, and mitochondrial biogenesis-related genes in porcine oocytes. These results indicated that melatonin promoted lipid metabolism and thereby provided an essential energy source for oocyte maturation and subsequent embryonic development. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Observations on the epidemiology of porcine parvovirus.

    Science.gov (United States)

    Johnson, R H; Donaldson-Wood, C; Allender, U

    1976-02-01

    Evidence presented suggests that porcine parvovirus is highly stable and infective. Introduction of virus to susceptible herds results in 100% infection rate within the following 3 months. Active immunity is associated with high persistent levels of haemagglutination-inhibitating (HI) antibody (greater than 256), piglets suckling immune sows acquiring HI titres between 10,000 and 40,000. Loss of passive immunity, measured by HI, occurs in a majority of pigs between 14 and 26 weeks of age (mean 21 weeks), whilst an average of 25% (2-47%) of pigs lose HI titres between 26 and 36 weeks of age. Susceptibility to challenge with virus does not occur until 3-5 weeks following loss of HI titres. In endemically infected herds 98-100% of adult pigs show serological evidence of active immunity. A significant proportion of gilts may not be actively immune to porcine parvovirus at the time of first service, and subsequent infection may occur while these gilts are pregnant.

  18. Phenol esterase activity of porcine skin.

    Science.gov (United States)

    Laszlo, Joseph A; Smith, Leslie J; Evans, Kervin O; Compton, David L

    2015-01-01

    The alkyl esters of plant-derived phenols may serve as slow-release sources for cutaneous delivery of antioxidants. The ability of skin esterases to hydrolyze phenolic esters was examined. Esters of tyrosol and hydroxytyrosol were prepared from decanoic and lipoic acids. Ferulic acid was esterified with octadecanol, glycerol, and dioleoylglycerol. These phenolic derivatives were treated in taurodeoxycholate microemulsion and unilamellar liposomes with ex vivo porcine skin and an aqueous extract of the skin. Extracted esterases hydrolyzed the microemulsions at rates in the order: tyrosyl lipoate > tyrosyl decanoate > hydroxytyrosyl lipoate > hydroxytyrosyl decanoate. The tyrosyl decanoate was subject to comparatively little hydrolysis (10-30% after 24h) when incorporated into liposomes, while hydroxytyrosyl decanoate in liposomes was not hydrolyzed at all by the skin extract. Ferulate esters were not hydrolyzed by the extract in aqueous buffer, microemulsion, nor liposomes. Tyrosyl decanoate applied topically to skin explants in microemulsion were readily hydrolyzed within 4h, while hydrolysis was minimal when applied in liposomes. These findings indicate that porcine skin displays a general esterase activity toward medium-chain esters of tyrosol and hydroxytyrosol, which can be moderated by the physiochemical properties of the lipid vehicle, but no feruloyl esterase activity. Published by Elsevier B.V.

  19. Genetic diversity of porcine Pasteurella multocida strains from the respiratory tract of healthy and diseased swine.

    Science.gov (United States)

    Bethe, Astrid; Wieler, Lothar H; Selbitz, Hans-J; Ewers, Christa

    2009-10-20

    A total of 382 porcine Pasteurella multocida strains, isolated from cases of pneumonia and progressive atrophic rhinitis (PAR) as well as from clinically healthy pigs of more than 150 German husbandries were characterized by detection of virulence-associated genes (VAGs) and ribotyping to understand the relationships between "commensal" and "pathogenic" strains, enabling a rational choice of vaccine strains. The diversity of the strains according to VAGs was low and mainly limited to capsular type genes (capA: 53.4%; capD: 45.8%; capF: 0.3%; cap-negative: 0.5%; hssB: 95.3%), dermonecrotoxin gene toxA (3.4%), as well as adhesion-related genes pfhaB (20.9%) and hgbB (84.3%). Ribotyping identified 13 patterns, but the vast majority of strains (95.8%) clustered in only three of these, namely IA-1 (45.5%), IA-7 (30.1%), and IIA-1 (20.2%). Pattern IA-1 was associated with capD(+) strains (93.6%) and harboured the majority of toxA(+) strains (84.6%). Pattern IA-7 mostly contained pfhaB(-), toxA(-)capA(+) strains (93.9%), while pattern IIA-1 was predominantly composed of pfhaB(+), toxA(-)capA(+) strains (87.0%). Clinical strains associated with pneumonia or PAR shared the above mentioned major ribotypes in comparable proportions with strains derived from healthy pigs, suggesting P. multocida to act more as an opportunistic than as an obligate pathogen in pigs. The limited number of subpopulations may either reflect a recent evolution of P. multocida in pigs or a selection by means of horizontal transfer of capsular genes, toxA or pfhaB. These data enforce further phylogenetic and epidemiological studies, examining the properties of different subpopulations of porcine P. multocida strains as well as factors of the porcine hosts themselves, which might be involved in disease susceptibility.

  20. Genetic Fusions of Heat-Labile Toxoid (LT) and Heat-Stable Toxin b (STb) of Porcine Enterotoxigenic Escherichia coli Elicit Protective Anti-LT and Anti-STb Antibodies ▿

    Science.gov (United States)

    Zhang, Weiping; Francis, David H.

    2010-01-01

    Enterotoxigenic Escherichia coli (ETEC)-associated diarrhea causes a substantial economic loss to swine producers worldwide. The majority of ETEC strains causing porcine diarrhea, especially postweaning diarrhea (PWD), produce heat-labile toxin (LT) and heat-stable toxin b (STb). LT is commonly used in vaccine development, but STb has not been included because of its poor immunogenicity. As a virulence factor in porcine diarrhea, STb needs to be included as an antigen for development of broad-spectrum vaccines. In this study, we used an LT toxoid (LTR192G [hereafter, LT192]) derived from porcine ETEC to carry a mature STb peptide for LT192-STb fusions to enhance STb immunogenicity for potential vaccine application. Anti-LT and anti-STb antibodies were detected in immunized rabbits and pigs. In addition, when challenged with an STb-positive ETEC strain, all 10 suckling piglets borne by immunized gilts remained healthy, whereas 7 out 9 piglets borne by unimmunized gilts developed moderate diarrhea. This study indicates that the LT192-STb fusion enhanced anti-STb immunogenicity and suggests the LT192-STb fusion antigen can be used in future vaccine development against porcine ETEC diarrhea. PMID:20505006

  1. Sequence conservation between porcine and human LRRK2

    DEFF Research Database (Denmark)

    Larsen, Knud; Madsen, Lone Bruhn

    2009-01-01

     Leucine-rich repeat kinase 2 (LRRK2) is a member of the ROCO protein superfamily (Ras of complex proteins (Roc) with a C-terminal Roc domain). Mutations in the LRRK2 gene lead to autosomal dominant Parkinsonism. We have cloned the porcine LRRK2 cDNA in an attempt to characterize conserved and th...... and expression patterns are conserved across species. The porcine LRRK2 gene was mapped to chromosome 5q25. The results obtained suggest that the LRRK2 gene might be of particular interest in our attempt to generate a transgenic porcine model for Parkinson's disease...

  2. A porcine model of haematogenous brain infectionwith staphylococcus aureus

    DEFF Research Database (Denmark)

    Astrup, Lærke Boye; Agerholm, Jørgen Steen; Nielsen, Ole Lerberg

    2012-01-01

    A PORCINE MODEL OF HAEMATOGENOUS BRAIN INFECTION WITH STAPHYLOCOCCUS AUREUS Astrup Lærke1, Agerholm Jørgen1, Nielsen Ole1, Jensen Henrik1, Leifsson Páll1, Iburg Tine2. 1: Faculty of Health and Medical Sciences, University of Copenhagen, Denmark boye@life.ku.dk 2: National Veterinary Institute......, Uppsala, Sweden Introduction Staphylococcus aureus (S.aureus) is a common cause of sepsis and brain abscesses in man and a frequent cause of porcine pyaemia. Here we present a porcine model of haematogenous S. aureus-induced brain infection. Materials and Methods Four pigs had two intravenous catheters...

  3. Reproductive Physiology of Marsupials

    Science.gov (United States)

    Sharman, G. B.

    1970-01-01

    Describes some unique features of marsupial reproduction which include (1) chromosomal sex determination, (2) reproductive system, (3) birth, (4) location, and (5) embryonic diapause. These features suggest that viviparity evolved separately in eutherian and marsupial stocks after their derivation from a common oviparous ancestor. Bibliography.…

  4. Multichannel Sound Reproduction

    Science.gov (United States)

    Pulkki, Ville

    Spatial reproduction of sound is a field in which the spatial attributes of a real recording room or a virtual space are reproduced to the listener. Spatial attributes include for example directions of sound sources, directions of reflections and envelopment by reverberation. Many such systems employ more than two loudspeakers to create virtual sources. This is called multichannel sound or spatial sound reproduction.

  5. The Reproduction of Intelligence

    Science.gov (United States)

    Meisenberg, Gerhard

    2010-01-01

    Although a negative relationship between fertility and education has been described consistently in most countries of the world, less is known about the relationship between intelligence and reproductive outcomes. Also the paths through which intelligence influences reproductive outcomes are uncertain. The present study uses the NLSY79 to analyze…

  6. Genetic Characterization of Porcine Circovirus Type 2 from Pigs with Porcine Circovirus Associated Diseases in Argentina

    Science.gov (United States)

    Pereda, Ariel; Piñeyro, Pablo; Bratanich, Ana; Quiroga, María Alejandra; Bucafusco, Danilo; Craig, María Isabel; Cappuccio, Javier; Machuca, Mariana; Rimondi, Agustina; Dibárbora, Marina; Sanguinetti, Hector Ramón; Perfumo, Carlos Juan

    2011-01-01

    Porcine circovirus type 2 (PCV-2) has been associated with syndromes grouped by the term porcine circovirus associated diseases (PCVAD). The PCV-2 isolates have been grouped into two major groups or genotypes according to their nucleotide sequence of whole genomes and/or ORF-2: PCV-2b, which have, in turn, been subdivided into three clusters (1A–1C), and PCV-2a, which has been subdivided into five clusters (2A–2E). In the present study, we obtained 16 sequences of PCV-2 from different farms from 2003 to 2008, from animals with confirmatory diagnosis of PCVAD. Since results showed an identity of 99.8% among them, they were grouped within a common cluster 1A-B. This preliminary study suggests a stable circulation of PCV-2b among the Argentinean pig population. PMID:23738099

  7. Genetic characterization of porcine circovirus type 2 from pigs with porcine circovirus associated diseases in Argentina.

    Science.gov (United States)

    Pereda, Ariel; Piñeyro, Pablo; Bratanich, Ana; Quiroga, María Alejandra; Bucafusco, Danilo; Craig, María Isabel; Cappuccio, Javier; Machuca, Mariana; Rimondi, Agustina; Dibárbora, Marina; Sanguinetti, Hector Ramón; Perfumo, Carlos Juan

    2011-01-01

    Porcine circovirus type 2 (PCV-2) has been associated with syndromes grouped by the term porcine circovirus associated diseases (PCVAD). The PCV-2 isolates have been grouped into two major groups or genotypes according to their nucleotide sequence of whole genomes and/or ORF-2: PCV-2b, which have, in turn, been subdivided into three clusters (1A-1C), and PCV-2a, which has been subdivided into five clusters (2A-2E). In the present study, we obtained 16 sequences of PCV-2 from different farms from 2003 to 2008, from animals with confirmatory diagnosis of PCVAD. Since results showed an identity of 99.8% among them, they were grouped within a common cluster 1A-B. This preliminary study suggests a stable circulation of PCV-2b among the Argentinean pig population.

  8. Reproductive health and justice.

    Science.gov (United States)

    Petchesky, R

    1993-01-01

    This article was based on a speech given in Rio de Janeiro in January 1994 at the Reproductive Health and Justice Conference. Questions were raised about the universality of reproductive rights. The suggestion was that Western norms and principles subordinated Southern meanings. A women's health advocate in Nigeria believed that poor and oppressed women were not able to consider limiting family size or to consider reproductive health when the critical concerns were health care, education, livelihood, and basic needs. Rights and needs go together. Reproductive and sexual rights must be understood in terms of social, economic, and political enabling conditions. The respect for women's bodily integrity and reproductive and sexual well-being was viewed as integral to being an effective social and political agent. Women group's have carved out distinct concepts of work, economic resources, education, and political empowerment. The differences in experiences between the North and the South must not be used to diminish the impact of population control forces and fundamentalists. Reproductive rights means giving women the power to make informed decisions about individual fertility, childrearing, and health and sexual activity and means the resources to make decisions effectively and safely. The origin of the definition must not be confused with the process of debate. Rights can be approached either as legal and formal entities and/or as political claims to change existing power structures. Reproductive rights when construed to be liberties or choices were viewed as ineffectual; the focus must be on gender, class, culture, ethnicity, and national needs. Social rights must be incorporated in the concept of reproductive rights and as such challenge structural adjustment programs that reduce expenditures on health and social services. Terminology that focused on "reproduction" obscured the larger focus on personal health and well being. The principles of reproductive rights

  9. Copper tolerance and virulence in bacteria

    Science.gov (United States)

    Ladomersky, Erik; Petris, Michael J.

    2015-01-01

    Copper (Cu) is an essential trace element for all aerobic organisms. It functions as a cofactor in enzymes that catalyze a wide variety of redox reactions due to its ability to cycle between two oxidation states, Cu(I) and Cu(II). This same redox property of copper has the potential to cause toxicity if copper homeostasis is not maintained. Studies suggest that the toxic properties of copper are harnessed by the innate immune system of the host to kill bacteria. To counter such defenses, bacteria rely on copper tolerance genes for virulence within the host. These discoveries suggest bacterial copper intoxication is a component of host nutritional immunity, thus expanding our knowledge of the roles of copper in biology. This review summarizes our current understanding of copper tolerance in bacteria, and the extent to which these pathways contribute to bacterial virulence within the host. PMID:25652326

  10. Riboregulators: Fine-Tuning Virulence in Shigella.

    Science.gov (United States)

    Fris, Megan E; Murphy, Erin R

    2016-01-01

    Within the past several years, RNA-mediated regulation (ribo-regulation) has become increasingly recognized for its importance in controlling critical bacterial processes. Regulatory RNA molecules, or riboregulators, are perpetually responsive to changes within the micro-environment of a bacterium. Notably, several characterized riboregulators control virulence in pathogenic bacteria, as is the case for each riboregulator characterized to date in Shigella. The timing of virulence gene expression and the ability of the pathogen to adapt to rapidly changing environmental conditions is critical to the establishment and progression of infection by Shigella species; ribo-regulators mediate each of these important processes. This mini review will present the current state of knowledge regarding RNA-mediated regulation in Shigella by detailing the characterization and function of each identified riboregulator in these pathogens.

  11. The expression of VEGF, myoglobin and CRP2 proteins regulating endometrial remodeling in the porcine endometrial tissues during follicular and luteal phase.

    Science.gov (United States)

    Lee, Seunghyung; Lee, Sang-Hee; Yang, Boo-Keun; Park, Choon-Keun

    2017-09-01

    Endometrial remodeling is important for successful embryo development and implantation in pigs. Therefore, this study investigated change of proteins regulating endometrial remodeling on follicular and luteal phase in porcine endometrial tissues. The endometrial tissue samples were collected from porcine uterus during follicular and luteal phase, vascular endothelial growth factor (VEGF), myoglobin and cysteine-rich protein 2 (CRP2) proteins were expressed by immnofluorescence, immunoblotting, and determined by 2-DE and MALDI-TOF/MS. We found that VEGF, myoglobin and CRP2 were strongly localized in endometrial tissues during luteal phase, but not follicular phase. The protein levels of VEGF, myoglobin and CRP2 in endometrial tissues were higher than luteal phase (P < 0.05). These results may provide understanding of intrauterine environment during estrous cycle in pigs, and will be used in animal reproduction for developing specific biomarkers in the future. © 2017 Japanese Society of Animal Science.

  12. Porcine aminopeptidase N mediated polarized infection by porcine epidemic diarrhea virus in target cells

    Energy Technology Data Exchange (ETDEWEB)

    Cong, Yingying; Li, Xiaoxue; Bai, Yunyun [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China); Lv, Xiaonan [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China); CAS Key Lab for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience & Technology of China, Beijing 100090 (China); Herrler, Georg [Institute for Virology, University of Veterinary Medicine, Hannover D-30559 (Germany); Enjuanes, Luis [Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC), Campus Universidad Autónoma de Madrid, Cantoblanco, Madrid (Spain); Zhou, Xingdong [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China); Qu, Bo [Faculty of Life Sciences, Northeast Agricultural University, Harbin 150030 (China); Meng, Fandan [Institute for Virology, University of Veterinary Medicine, Hannover D-30559 (Germany); Cong, Chengcheng [College Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161 (China); Ren, Xiaofeng; Li, Guangxing [College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030 (China)

    2015-04-15

    Infection of polarized intestinal epithelial cells by porcine epidemic diarrhea virus (PEDV) was characterized. Indirect immunofluorescence assay, real-time PCR, and transmission electron microscopy confirmed PEDV can be successfully propagated in immortalized swine small intestine epithelial cells (IECs). Infection involved porcine aminpeptidase N (pAPN), a reported cellular receptor for PEDV, transient expression of pAPN and siRNA targeted pAPN increased and decreased the infectivity of PEDV in IECs, respectively. Subsequently, polarized entry into and release from both Vero E6 and IECs was analyzed. PEDV entry into polarized cells and pAPN grown on membrane inserts occurs via apical membrane. The progeny virus released into the medium was also quantified which demonstrated that PEDV is preferentially released from the apical membrane. Collectively, our data demonstrate that pAPN, the cellular receptor for PEDV, mediates polarized PEDV infection. These results imply the possibility that PEDV infection may proceed by lateral spread of virus in intestinal epithelial cells. - Highlights: • PEDV infection of polarized intestinal epithelial cells (IECs) was characterized. • Porcine aminpeptidase N (pAPN) facilitated PEDV infection in IECs. • PEDV entry into and release from polarized cell via its apical membrane. • PEDV infection may proceed by lateral spread of virus in IECs.

  13. Rapamycin-induced autophagy restricts porcine epidemic diarrhea virus infectivity in porcine intestinal epithelial cells.

    Science.gov (United States)

    Ko, Seongyeol; Gu, Min Jeong; Kim, Cheol Gyun; Kye, Yoon Chul; Lim, Younggap; Lee, Ji Eun; Park, Byung-Chul; Chu, Hyuk; Han, Seung Hyun; Yun, Cheol-Heui

    2017-10-01

    Porcine epidemic diarrhea virus (PEDV) invades porcine intestinal epithelial cells (IECs) and causes diarrhea and dehydration in pigs. In the present study, we showed a suppression of PEDV infection in porcine jejunum intestinal epithelial cells (IPEC-J2) by an increase in autophagy. Autophagy was activated by rapamycin at a dose that does not affect cell viability and tight junction permeability. The induction of autophagy was examined by LC3I/LC3II conversion. To confirm the autophagic-flux (entire autophagy pathway), autophagolysosomes were examined by an immunofluorescence assay. Pre-treatment with rapamycin significantly restricted not only a 1 h infection but also a longer infection (24 h) with PEDV, while this effect disappeared when autophagy was blocked. Co-localization of PEDV and autophagosomes suggests that PEDV could be a target of autophagy. Moreover, alleviation of PEDV-induced cell death in IPEC-J2 cells pretreated with rapamycin demonstrates a protective effect of rapamycin against PEDV-induced epithelial cell death. Collectively, the present study suggests an early prevention against PEDV infection in IPEC-J2 cells via autophagy that might be an effective strategy for the restriction of PEDV, and opens up the possibility of the use of rapamycin in vivo as an effective prophylactic and prevention treatment. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Interphase chromosome positioning in in vitro porcine cells and ex vivo porcine tissues

    Directory of Open Access Journals (Sweden)

    Foster Helen A

    2012-11-01

    Full Text Available Abstract Background In interphase nuclei of a wide range of species chromosomes are organised into their own specific locations termed territories. These chromosome territories are non-randomly positioned in nuclei which is believed to be related to a spatial aspect of regulatory control over gene expression. In this study we have adopted the pig as a model in which to study interphase chromosome positioning and follows on from other studies from our group of using pig cells and tissues to study interphase genome re-positioning during differentiation. The pig is an important model organism both economically and as a closely related species to study human disease models. This is why great efforts have been made to accomplish the full genome sequence in the last decade. Results This study has positioned most of the porcine chromosomes in in vitro cultured adult and embryonic fibroblasts, early passage stromal derived mesenchymal stem cells and lymphocytes. The study is further expanded to position four chromosomes in ex vivo tissue derived from pig kidney, lung and brain. Conclusions It was concluded that porcine chromosomes are also non-randomly positioned within interphase nuclei with few major differences in chromosome position in interphase nuclei between different cell and tissue types. There were also no differences between preferred nuclear location of chromosomes in in vitro cultured cells as compared to cells in tissue sections. Using a number of analyses to ascertain by what criteria porcine chromosomes were positioned in interphase nuclei; we found a correlation with DNA content.

  15. Preliminary evaluation of porcine zona pellucida (PZP) immunocontraception for behavioral effects in feral horses (Equus caballus).

    Science.gov (United States)

    Powell, D M

    1999-01-01

    Successful management of captive populations of wild animals requires effective control of reproduction. Contraception is one tool for controlling reproduction of animals in zoos; however, the options available to the animal manager are limited. Contraceptives vary in efficacy, reversibility, and side effects, and thus may not be suitable for widespread use. One consideration when selecting a contraceptive is its potential for side effects on behavior, especially given the fact that reproduction plays such a prominent role in the biology of any species. To date, there have been few evaluations of contraceptives for behavioral effects, and those that have been conducted have focused on hormone-based contraceptives. This study sought to evaluate a novel method of population control, immunocontraception, for behavioral effects in a population of feral horses. Porcine zona pellucida (PZP) immunocontraception prevents fertilization of ova and does not alter normal hormone secretion patterns. It therefore should leave the animal behaviorally intact in terms of reproductive behavior. The study examined the behavior of 43 sexually mature mares on Assateague Island during the 1997 breeding season and, with help from Earthwatch volunteers, collected observations over a 3-month period. The study found no significant differences between treated and untreated mares in general activity budget, aggression given or received, and spatial relationships relative to the stallion. These preliminary findings indicate that PZP contraception seems to have no acute behavioral effects on the behavior of individuals. The study findings also suggest that PZP could be a desirable and effective management tool for captive species in which social behavior plays an integral role in group dynamics. Analyses of group level effects and population level effects are continuing.

  16. Helicobacter pylori virulence factors in gastric carcinogenesis

    OpenAIRE

    Wen, Sicheng; Moss, Steven F.

    2008-01-01

    Helicobacter pylori infection is the most important risk factor in the development of non-cardia gastric adenocarcinoma; host genetic variability and dietary co-factors also modulate risk. Because most H. pylori infections do not cause cancer, H. pylori heterogeneity has been investigated to identify possible virulence factors. The strongest candidates are genes within the cag (cytotoxin associated antigen) pathogenicity island, including the gene encoding the CagA protein, as well as polymor...

  17. Surgical induction of choroidal neovascularization in a porcine model

    DEFF Research Database (Denmark)

    Lassota, Nathan; Kiilgaard, Jens Folke; Prause, Jan Ulrik

    2007-01-01

    PURPOSE: To develop a reproducible surgical technique for the induction of choroidal neovascularization (CNV) in the subretinal space of porcine eyes and to analyse the resulting CNV clinically and histologically. METHODS: Two different modifications of a surgical technique previously described...

  18. Porcine radial artery decellularization by high hydrostatic pressure.

    Science.gov (United States)

    Negishi, Jun; Funamoto, Seiichi; Kimura, Tsuyoshi; Nam, Kwangoo; Higami, Tetsuya; Kishida, Akio

    2015-11-01

    Many types of decellularized tissues have been studied and some have been commercially used in clinics. In this study, small-diameter vascular grafts were made using HHP to decellularize porcine radial arteries. One decellularization method, high hydrostatic pressure (HHP), has been used to prepare the decellularized porcine tissues. Low-temperature treatment was effective in preserving collagen and collagen structures in decellularized porcine carotid arteries. The collagen and elastin structures and mechanical properties of HHP-decellularized radial arteries were similar to those of untreated radial arteries. Xenogeneic transplantation (into rats) was performed using HHP-decellularized radial arteries and an untreated porcine radial artery. Two weeks after transplantation into rat carotid arteries, the HHP-decellularized radial arteries were patent and without thrombosis. In addition, the luminal surface of each decellularized artery was covered by recipient endothelial cells and the arterial medium was fully infiltrated with recipient cells. Copyright © 2012 John Wiley & Sons, Ltd.

  19. Acute Simvastatin Inhibits KATP Channels of Porcine Coronary Artery Myocytes

    National Research Council Canada - National Science Library

    Seto, Sai Wang; Au, Alice Lai Shan; Poon, Christina Chui Wa; Zhang, Qian; Li, Rachel Wai Sum; Yeung, John Hok Keung; Kong, Siu Kai; Ngai, Sai Ming; Wan, Song; Ho, Ho Pui; Lee, Simon Ming Yuen; Hoi, Maggie Pui Man; Chan, Shun Wan; Leung, George Pak Heng; Kwan, Yiu Wa

    2013-01-01

    ...]o uptake measurements. Results The cromakalim (10 nM to 10 µM)- and pinacidil (10 nM to 10 µM)-induced concentration-dependent relaxation of porcine coronary artery was inhibited by simvastatin...

  20. A comparative anatomic and physiologic overview of the porcine heart

    National Research Council Canada - National Science Library

    Lelovas, Pavlos P; Kostomitsopoulos, Nikolaos G; Xanthos, Theodoros T

    2014-01-01

    .... The porcine heart bears a close resemblance to the human heart in terms of its coronary circulation and hemodynamic similarities and offers ease of implementation of methods and devices from human healthcare facilities...

  1. Purification and characterization of acylation stimulating protein from porcine serum.

    Science.gov (United States)

    Zhang, Hong; Jacobi, Sheila K; Toombs, Candice F; Cianflone, Katherine H; Nersesian, Natalya; Sarath, Gautam; Miner, Jess L

    2002-07-01

    A method for purifying acylation stimulating protein (ASP) from porcine serum is described. The mRNA encoding ASP was cloned by reverse transcriptase-polymerase chain reaction which predicted a 76 residue peptide. Based on this sequence, we generated antisera to a C-terminal peptide (ASP(1-20)) which aided ASP purification. Identity of the purified protein was verified by N-terminal sequencing. The molecular mass of porcine ASP is 8926. Porcine ASP stimulated esterification of fatty acid into triacylglycerol in cultured human cells with potency similar to that of human ASP (twofold at 5 microM). Based on this evidence that ASP exists in porcine blood, and that it has acylation stimulating activity, we propose that ASP may play a role in regulation of energy storage in adipose tissue in the pig.

  2. The future of human reproduction

    National Research Council Canada - National Science Library

    Overall, Christine

    1989-01-01

    ... Contradictions III SOCIAL POLICY QUESTIONS Pregnancy as Justification for Loss of Juridical Autonomy Sanda Rodgers 174 Prenatal Diagnosis: Reproductive Choice? Reproductive Control? Abby Lippman ...

  3. The politics of reproduction.

    Science.gov (United States)

    Ginsburg, F; Rapp, R

    1991-01-01

    The topic of human reproduction encompasses events throughout the human and especially female life-cycle as well as ideas and practices surrounding fertility, birth, and child care. Most of the scholarship on the subject, up through the 1960s, was based on cross-cultural surveys focused on the beliefs, norms, and values surrounding reproductive behaviors. Multiple methodologies and subspecialties, and fields like social history, human biology, and demography were utilized for the analysis. The concept of the politics of reproduction synthesizes local and global perspectives. The themes investigated include: the concept of reproduction, population control, and the internationalization of state and market interests (new reproductive technologies); social movements and contested domains; medicalization and its discontents; fertility and its control; adolescence and teen pregnancy; birth; birth attendants; the construction of infancy and the politics of child survival; rethinking the demographic transition; networks of nurturance; and meanings of menopause. The medicalization of reproduction is a central issue of studies of birth, midwifery, infertility, and reproductive technologies. Scholars have also analyzed different parts of the female life-cycle as medical problems. Other issues worth analysis include the internationalization of adoption and child care workers; the crisis of infertility of low-income and minority women who are not candidates for expensive reproductive technologies; the concerns of women at high risk for HIV whose cultural status depends on their fertility; questions of reproduction concerning, lesbians and gay men (artificial insemination and discrimination in child rearing); the study of menopause; and fatherhood. New discourse analysis is used to analyze state eugenic policies; conflicts over Western neocolonial influences in which women's status as childbearers represent nationalist interests; fundamentalist attacks on abortion rights; and

  4. Tiamulin resistance in porcine Brachyspira pilosicoli isolates.

    Science.gov (United States)

    Pringle, M; Landén, A; Franklin, A

    2006-02-01

    There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified.

  5. A Novel Porcine Graft for Regeneration of Bone Defects

    Directory of Open Access Journals (Sweden)

    Eisner Salamanca

    2015-05-01

    Full Text Available Bone regeneration procedures require alternative graft biomaterials to those for autogenous bone. Therefore, we developed a novel porcine graft using particle sizes of 250–500 μm and 500–1000 μm in rabbit calvarial bone defects and compared the graft properties with those of commercial hydroxyapatite (HA/beta-tricalcium phosphate (β-TCP over eight weeks. Surgery was performed in 20 adult male New Zealand white rabbits. During a standardized surgical procedure, four calvarial critical-size defects of 5 mm diameter and 3 mm depth were prepared. The defects were filled with HA/β-TCP, 250–500 μm or 500–1000 μm porcine graft, and control defects were not filled. The animals were grouped for sacrifice at 1, 2, 4, and 8 weeks post-surgery. Subsequently, sample blocks were prepared for micro-computed tomography (micro-CT scanning and histological sectioning. Similar bone formations were observed in all three treatment groups, although the 250–500 μm porcine graft performed slightly better. Rabbit calvarial bone tissue positively responded to porcine grafts and commercial HA/β-TCP, structural analyses showed similar crystallinity and porosity of the porcine and HA/β-TCP grafts, which facilitated bone formation through osteoconduction. These porcine grafts can be considered as graft substitutes, although further development is required for clinical applications.

  6. Expression of bioactive porcine interferon-alpha in Lactobacillus casei.

    Science.gov (United States)

    Ma, Shi-jie; Li, Kun; Li, Xin-Sheng; Guo, Xiao-Qing; Fu, Peng-Fei; Yang, Ming-Fan; Chen, Hong-Ying

    2014-09-01

    In this study, we constructed an expression cassette containing the inducible lac promoter and the secretion signal from an S-layer protein of Lactobacillus brevis for the expression of porcine interferon-alpha (IFN-α) in Lactobacillus casei (Lb. casei). Reverse-transcriptase PCR verified the presence of porcine IFN-α mRNA in the recombinant Lb. casei. The porcine IFN-α protein expressed in the recombinant Lb. casei was identified by both Western blot analysis and ELISA. We used various pH values and induction times to optimize the yield of IFN-α, and found that induction with 0.8% lactose for 16 h under anaerobic conditions produced the highest concentrations of IFN-α. Furthermore, the activity of porcine IFN-α in the cultural supernatant was evaluated on ST cells infected with pseudorabies virus. The results revealed that porcine IFN-α inhibited virus replication in vitro. The findings of our study indicate that recombinant Lb. casei producing porcine IFN-α has great potential for use as a novel oral antiviral agent in animal healthcare.

  7. Porcine bioengineered scaffolds as new frontiers in regenerative medicine.

    Science.gov (United States)

    Park, K M; Woo, H M

    2012-05-01

    Porcine organs are attractive for xenotransplantation, if severe immunologic concerns can be overcome. Recently, reengineered organs, with heterologous cellular materials removed but preserved organ architecture and vasculature have been created using small rodents in an effort to produce customized bioengineered organs. However, few studies have been performed to generate bioengineered organs from porcine sources. The aim of this work was to produce 3-D bioengineered scaffolds from major porcine organs, preserving the native morphology and vascular structures with complete removal of cellular and nuclear materials. We decellularized porcine heart, liver, and kidney using a peristaltic pump system with 1% sodium dodecyl sulfate. The preservation of major architecture and vasculature was confirmed by gross findings, ultrasonography, and angiography. Hematoxylin and eosin staining revealed no evidence of nuclear or cytoplasmic residues. Quantitative DNA analysis demonstrated a substantial reduction (0%-8%) of porcine DNA in the scaffolds. These results suggested that 3-D bioengineered scaffolds of porcine organs may have tremendous potential to produce non-immunogenic transplantable organs as well as beneficial tools for biomedical studies on organ re-engineering and repair. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Regional variations in the histology of porcine skin.

    Science.gov (United States)

    Turner, Neill J; Pezzone, Dominic; Badylak, Stephen F

    2015-04-01

    Porcine skin is commonly used as a model for human skin injury and as a source material for biologic scaffold materials. Although remarkable similarities between porcine and human skin exist, regional anatomic variations present in human skin are also present in porcine skin. The objective of this study was to evaluate the structure of porcine skin from 11 different anatomic regions in the American Yorkshire crossbreed. Both qualitative and quantitative methods were used, with emphasis on epidermal and dermal thickness, hair follicle density, and collagen and elastin composition and distribution. The results showed that significant regional differences in skin histology exist, particularly with regard to the thickness of the dermis and epidermis and the amount of collagen and elastin within each tissue. Differences were also seen in the distribution of type I and type III collagen within the dermis. Therefore, while porcine skin shares many similarities with human skin, distinct regional differences in composition and morphology exist. This study highlights the importance of appreciating these regional differences to avoid misinterpretation of experimental results when using porcine skin as a human analogue.

  9. How Active Are Porcine Endogenous Retroviruses (PERVs?

    Directory of Open Access Journals (Sweden)

    Joachim Denner

    2016-08-01

    Full Text Available Porcine endogenous retroviruses (PERVs represent a risk factor if porcine cells, tissues, or organs were to be transplanted into human recipients to alleviate the shortage of human transplants; a procedure called xenotransplantation. In contrast to human endogenous retroviruses (HERVs, which are mostly defective and not replication-competent, PERVs are released from normal pig cells and are infectious. PERV-A and PERV-B are polytropic viruses infecting cells of several species, among them humans; whereas PERV-C is an ecotropic virus infecting only pig cells. Virus infection was shown in co-culture experiments, but also in vivo, in the pig, leading to de novo integration of proviruses in certain organs. This was shown by measurement of the copy number per cell, finding different numbers in different organs. In addition, recombinations between PERV-A and PERV-C were observed and the recombinant PERV-A/C were found to be integrated in cells of different organs, but not in the germ line of the animals. Here, the evidence for such in vivo activities of PERVs, including expression as mRNA, protein and virus particles, de novo infection and recombination, will be summarised. These activities make screening of pigs for provirus number and PERV expression level difficult, especially when only blood or ear biopsies are available for analysis. Highly sensitive methods to measure the copy number and the expression level will be required when selecting pigs with low copy number and low expression of PERV as well as when inactivating PERVs using the clustered regularly interspaced short palindromic repeats (CRISPR/CRISPR-associated nuclease (CRISPR/Cas technology.

  10. Porcine circovirus diseases: a review of PMWS.

    Science.gov (United States)

    Baekbo, P; Kristensen, C S; Larsen, L E

    2012-03-01

    This article is a review on post-weaning multisystemic wasting syndrome (PMWS), the first described disease among the porcine circovirus diseases (PCVD). Post-weaning multisystemic wasting syndrome has, since its appearance in Canada in 1991, been seen in all major pig producing countries. To diagnose PMWS at herd level typical clinical appearance consisting of wasting and increased mortality must be combined with finding at autopsy of diseased pigs, where typical microscopic findings in the lymphatic tissue must be present. Post-weaning multisystemic wasting syndrome significantly increases the mortality and reduces the daily weight gain in weaner pig and/or in finishing pigs. Post-weaning multisystemic wasting syndrome can be transmitted by pig-to-pig contact and some studies point at airborne transmission as a possibility. Studies in Europe have shown several risk factors that either increase or decrease the risk for a pig herd to be affected by PMWS. At the pig level, studies have shown the importance of maternal immunity as protection for subsequent development of PMWS. To control PMWS, good production management and control of other diseases are crucial. Since 2004, commercial vaccines against Porcine Circo Virus type 2 have been coming on the market and many studies have shown great benefits of these to control PMWS. Today, sow vaccines as well as piglet vaccines are available in most countries. An extensive meta-analysis of many of the vaccines has shown a comparable good efficacy of the vaccines in significantly reducing mortality and increasing weight gain of the pigs. © 2012 Blackwell Verlag GmbH.

  11. Cod reproductive ecology

    DEFF Research Database (Denmark)

    Røjbek, Maria

    reproduction. The overall objective of the thesis is to investigate the role of lipids in reproduction of cod (Gadus morhua) in the Central Baltic Sea. The first objective is to examine the seasonal variation in content of lipid and EFA in whole prey species of cod (Paper I). The second objective...... is to investigate the variation in lipid content, EFA and antioxidants of female Baltic cod gonads and livers during the reproductive cycle (Paper II) and to examine whether there is a deficiency in lipid energy and dietary EFA that could explain the delayed spawning time observed in the Baltic cod (Paper III...

  12. Transcription analysis of the responses of porcine heart to Erysipelothrix rhusiopathiae.

    Science.gov (United States)

    Kang, Chao; Zhang, Qiang; Zhu, Weifeng; Cai, Chengzhi; Sun, Xiaomei; Jin, Meilin

    2017-01-01

    Erysipelothrix rhusiopathiae (E. rhusiopathiae) is the causative agent of swine erysipelas. This microbe has caused great economic losses in China and in other countries. In this study, high-throughput cDNA microarray assays were employed to evaluate the host responses of porcine heart to E. rhusiopathiae and to gain additional insights into its pathogenesis. A total of 394 DE transcripts were detected in the active virulent E. rhusiopathiae infection group compared with the PBS group at 4 days post-infection. Moreover, 262 transcripts were upregulated and 132 transcripts were downregulated. Differentially expressed genes were involved in many vital functional classes, including inflammatory and immune responses, signal transduction, apoptosis, transport, protein phosphorylation and dephosphorylation, metabolic processes, chemotaxis, cell adhesion, and innate immune responses. Pathway analysis demonstrated that the most significant pathways were Chemokine signaling pathway, NF-kappa B signaling pathway, TLR pathway, CAMs, systemic lupus erythematosus, chemokine signaling pathway, Cytokine-cytokine receptor interaction, PI3K-Akt signaling pathway, Phagosome, HTLV-I infection, Measles, Rheumatoid arthritis and natural-killer-cell-mediated cytotoxicity. The reliability of our microarray data was verified by performing quantitative real-time PCR. This study is the first to document the response of piglet heart to E. rhusiopathiae infection. The observed gene expression profile could help screen potential host agents that can reduce the prevalence of E. rhusiopathiae. The profile might also provide insights into the underlying pathological changes that occur in pigs infected with E. rhusiopathiae.

  13. Effects of Actinobacillus pleuropneumoniae cytotoxins on generation of oxygen radicals by porcine neutrophils

    Directory of Open Access Journals (Sweden)

    Simson Tarigan

    1999-03-01

    Full Text Available Cytotoxins produced by Actinobacillus pleuropneumoniae (App suggested to be the most important pathogenic and virulent factors for this organism. However, the mechanisms on how the cytotoxins contribute to the disease process remain unclear. The purpose of this study is to investigate the effect of the cytotoxins on the oxidative-burst metabolism of porcine neutrophils. In this study, neutrophils were firstly loaded with an oxidative probe dichlorofluorescin diacetate (DCFHDA then expose to cytotoxins. Cells producing oxygen radicals emitted fluorescence and its intensity was measured with a FACScan flow cytometer. All cytotoxins derived from either App serotypes producing ApxI and ApxII, App serotypes producing ApxII only, or App serotypes producing ApxII and ApxIII were capable of stimulating neutrophils for oxygen-radical generation. However, compared with phorbol myristate acetate (PMA, App cytotoxins were much weaker as stimulants for oxygen radicals. In addition, Apx preparation stimulated an oxidative-burst metabolism of neutrophils at a low, narrow range of Apx doses. At higher doses, the toxins inhibit the oxidative burst metabolism. The effects of cytotoxins produced by App during infection on recruited neutrophils into the lungs are assumed to be comparable to those observed in this in vitro study. Neutrophils, and other host cells, adjacent to the bacteria become lysis due to high toxin concentration, whereas those at some distance to the bacteria produce oxygen radicals which in turn cause tissue damage or necrosis.

  14. The effect of depopulation and restocking on reproductive and growth performances on Japanese commercial swine farms.

    Science.gov (United States)

    Sasaki, Yosuke; Sekiguchi, Satoshi; Uemura, Ryoko; Sueyoshi, Masuo

    2016-02-01

    This study compared the reproductive and growth performances of pigs before and after depopulation and restocking after a foot-and-mouth disease outbreak in Japan. Data for the time period before and after depopulation and restocking were obtained from three farrow-to-finish farms. As a result of depopulation and restocking, hygiene levels were improved, and common infectious diseases, such as porcine reproductive and respiratory syndrome and Aujeszky's disease, remained undetected on the farms. Compared with before depopulation, reproductive and growth performances improved after depopulation; the number of total pigs born was higher, the postweaning mortality rate was lower, and the age at slaughter was lower (Preproductive and growth performances of pigs.

  15. Bacterial virulence in the moonlight: multitasking bacterial moonlighting proteins are virulence determinants in infectious disease.

    Science.gov (United States)

    Henderson, Brian; Martin, Andrew

    2011-09-01

    Men may not be able to multitask, but it is emerging that proteins can. This capacity of proteins to exhibit more than one function is termed protein moonlighting, and, surprisingly, many highly conserved proteins involved in metabolic regulation or the cell stress response have a range of additional biological actions which are involved in bacterial virulence. This review highlights the multiple roles exhibited by a range of bacterial proteins, such as glycolytic and other metabolic enzymes and molecular chaperones, and the role that such moonlighting activity plays in the virulence characteristics of a number of important human pathogens, including Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Helicobacter pylori, and Mycobacterium tuberculosis.

  16. The phytoplasmal virulence factor TENGU causes plant sterility by downregulating of the jasmonic acid and auxin pathways.

    Science.gov (United States)

    Minato, Nami; Himeno, Misako; Hoshi, Ayaka; Maejima, Kensaku; Komatsu, Ken; Takebayashi, Yumiko; Kasahara, Hiroyuki; Yusa, Akira; Yamaji, Yasuyuki; Oshima, Kenro; Kamiya, Yuji; Namba, Shigetou

    2014-12-10

    Despite plants infected by pathogens are often unable to produce offspring, it remains unclear how sterility is induced in host plants. In this study, we demonstrate that TENGU, a phytoplasmal virulence peptide known as a dwarfism inducer, acts as an inducer of sterility. Transgenic expression of TENGU induced both male and female sterility in Arabidopsis thaliana flowers similar to those observed in double knockout mutants of auxin response factor 6 (ARF6) and ARF8, which are known to regulate floral development in a jasmonic acid (JA)-dependent manner. Transcripts of ARF6 and ARF8 were significantly decreased in both tengu-transgenic and phytoplasma-infected plants. Furthermore, JA and auxin levels were actually decreased in tengu-transgenic buds, suggesting that TENGU reduces the endogenous levels of phytohormones by repressing ARF6 and ARF8, resulting in impaired flower maturation. TENGU is the first virulence factor with the effects on plant reproduction by perturbation of phytohormone signaling.

  17. Reproductive prognosis in endometriosis

    DEFF Research Database (Denmark)

    Hjordt Hansen, Maj V; Dalsgaard, Torur; Hartwell, Dorthe

    2014-01-01

    OBJECTIVE: To assess the reproductive long-term prognosis of women with and without endometriosis, to explore changes over time, and to quantify the contribution of artificial reproductive techniques. DESIGN: Cohort study. SETTING: Denmark 1977-2009. SAMPLE: Data retrieved from four national...... registries. Among 15-49-year-old women during the period 1977-82, 24 667 were diagnosed with endometriosis and 98 668 (1:4) women without endometriosis were age-matched. METHODS: To assess long-term reproductive prognosis, all pregnancy outcomes were identified among the women with and without endometriosis......, but this was restricted to pregnancies from assisted reproduction. CONCLUSION: Women with endometriosis have slightly fewer children, but this lessened over time due to artificially conceived pregnancies. The risk for miscarriages and ectopic pregnancies was increased compared with women without the disease....

  18. Selective Reproductive Technologies

    DEFF Research Database (Denmark)

    Gammeltoft, Tine; Wahlberg, Ayo

    2014-01-01

    From a historical perspective, selective reproduction is nothing new. Infanticide, abandonment, and selective neglect of children have a long history, and the widespread deployment of sterilization and forced abortion in the twentieth century has been well documented. Yet in recent decades...... selective reproduction has been placed under the aegis of science and expertise in novel ways. New laboratory and clinical techniques allow for the selective fertilization of gametes, implantation of embryos, or abortion of fetuses. Although they will often overlap with assisted reproductive technologies...... (ARTs), what we term selective reproductive technologies (SRTs) are of a more specific nature: Rather than aiming to overcome infertility, they are used to prevent or allow the birth of certain kinds of children. This review highlights anthropological research into SRTs in different parts of the world...

  19. Reproductive and developmental toxicology

    National Research Council Canada - National Science Library

    Gupta, Ramesh C

    2011-01-01

    .... With a special focus on placental toxicity, this book is the only available reference to connect the three key risk stages, and is the only resource to include reproductive and developmental toxicity in domestic animals, fish, and wildlife.

  20. Reproductive data for groundfish

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The ROCKFISH database houses data from rockfish species collected by the SWFSC FED along the California coast as part of a reproductive study originating in the...

  1. Avian reproductive physiology

    Science.gov (United States)

    Gee, G.F.; Gibbons, Edward F.; Durrant, Barbara S.; Demarest, Jack

    1995-01-01

    Knowledge of the many physiological factors associated with egg production , fertility, incubation, and brooding in nondomestic birds is limited. Science knows even less about reproduction in most of the 238 endangered or threatened birds. This discussion uses studies of nondomestic and, when necessary, domestic birds to describe physiological control of reproduction. Studies of the few nondomestic avian species show large variation in physiological control of reproduction. Aviculturists, in order to successfully propagate an endangered bird, must understand the bird's reproductive peculiarities. First, investigators can do studies with carefully chosen surrogate species, but eventually they need to confirm the results in the target endangered bird. Studies of reproduction in nondomestic birds increased in the last decade. Still, scientists need to do more comparative studies to understand the mechanisms that control reproduction in birds. New technologies are making it possible to study reproductive physiology of nondomestic species in less limiting ways. These technologies include telemetry to collect information without inducing stress on captives (Howey et al., 1987; Klugman, 1987), new tests for most of the humoral factors associated with reproduction, and the skill to collect small samples and manipulate birds without disrupting the physiological mechanisms (Bercovitz et al., 1985). Managers are using knowledge from these studies to improve propagation in zoological parks, private and public propagation facilities, and research institutions. Researchers need to study the control of ovulation, egg formation, and oviposition in the species of nondomestic birds that lay very few eggs in a season, hold eggs in the oviduct for longer intervals, or differ in other ways from the more thoroughly studied domestic birds. Other techniques that would enhance propagation for nondomestlc birds include tissue culture of cloned embryonic cells, cryopreservation of embryos

  2. Reproductive Liberty and Overpopulation

    OpenAIRE

    Carol A. Kates

    2004-01-01

    Despite substantial evidence pointing to a looming Malthusian catastrophe, governmental measures to reduce population have been opposed both by religious conservatives and by many liberals, especially liberal feminists. Liberal critics have claimed that 'utilitarian' population policies violate a 'fundamental right of reproductive liberty'. This essay argues that reproductive liberty should not be considered a fundamental human right, or certainly not an indefeasible right. It should, instead...

  3. Thyroid and male reproduction

    Directory of Open Access Journals (Sweden)

    Anand Kumar

    2014-01-01

    Full Text Available Male reproduction is governed by the classical hypothalamo-hypophyseal testicular axis: Hypothalamic gonadotropin releasing hormone (GnRH, pituitary luteinizing hormone (LH and follicle stimulating hormone (FSH and the gonadal steroid, principally, testosterone. Thyroid hormones have been shown to exert a modulatory influence on this axis and consequently the sexual and spermatogenic function of man. This review will examine the modulatory influence of thyroid hormones on male reproduction.

  4. Thyroid and male reproduction.

    Science.gov (United States)

    Kumar, Anand; Shekhar, Skand; Dhole, Bodhana

    2014-01-01

    Male reproduction is governed by the classical hypothalamo-hypophyseal testicular axis: Hypothalamic gonadotropin releasing hormone (GnRH), pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) and the gonadal steroid, principally, testosterone. Thyroid hormones have been shown to exert a modulatory influence on this axis and consequently the sexual and spermatogenic function of man. This review will examine the modulatory influence of thyroid hormones on male reproduction.

  5. Pseudomonas aeruginosa Virulence Analyzed in a Dictyostelium discoideum Host System

    OpenAIRE

    Cosson, Pierre; Zulianello, Laurence; Join-Lambert, Olivier; Faurisson, François; Gebbie, Leigh; Benghezal, Mohammed; Van Delden, Christian; Kocjancic Curty, Lasta; Köhler, Thilo

    2002-01-01

    Pseudomonas aeruginosa is an important opportunistic pathogen that produces a variety of cell-associated and secreted virulence factors. P. aeruginosa infections are difficult to treat effectively because of the rapid emergence of antibiotic-resistant strains. In this study, we analyzed whether the amoeba Dictyostelium discoideum can be used as a simple model system to analyze the virulence of P. aeruginosa strains. The virulent wild-type strain PAO1 was shown to inhibit growth of D. discoide...

  6. Unravelling the contribution of the Penicillium expansum PeSte12 transcription factor to virulence during apple fruit infection.

    Science.gov (United States)

    Sánchez-Torres, Paloma; Vilanova, Laura; Ballester, Ana Rosa; López-Pérez, Mario; Teixidó, Neus; Viñas, Inmaculada; Usall, Josep; González-Candelas, Luis; Torres, Rosario

    2018-02-01

    Blue mould disease caused by Penicillium expansum infection is one of the most important diseases of pome fruit accounting for important economic losses. In the present study, the PeSte12 transcription factor gene was identified, and deletant mutants were produced by gene replacement. Knockout mutants showed a significant decrease of virulence during apple fruit infection. Virulence was affected by the maturity stage of the fruit (immature, mature and over-mature), and disease severity was notably reduced when the apples were stored at 0 °C. The ΔPeSte12 mutants resulted defective in asexual reproduction, producing less conidia, but this characteristic did not correlate with differences in microscopic morphology. In addition, the ΔPeSte12 mutants produced higher quantity of hydrogen peroxide than the wild type strain. Gene expression analysis revealed that PeSte12 was induced over time during apple infection compared to axenic growth, particularly from 2 dpi, reinforcing its role in virulence. Analysis of transcriptional abundance of several genes in ΔPeSte12 mutants showed that in most of the evaluated genes, PeSte12 seemed to act as a negative regulator during axenic growth, as most of them exhibited an increasing expression pattern along the time period evaluated. The highest expression values corresponded to detoxification, ATPase activity, protein folding and basic metabolism. Gene expression analysis during apple infection showed that 3 out of 9 analysed genes were up regulated; thus, PeSte12 seemed to exert a positive control to particular type of aldolase. These results demonstrate the PeSte12 transcription factor could play an important role in P. expansum's virulence and asexual reproduction. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Asexual Reproduction in Holothurians

    Science.gov (United States)

    Dolmatov, Igor Yu.

    2014-01-01

    Aspects of asexual reproduction in holothurians are discussed. Holothurians are significant as fishery and aquaculture items and have high commercial value. The last review on holothurian asexual reproduction was published 18 years ago and included only 8 species. An analysis of the available literature shows that asexual reproduction has now been confirmed in 16 holothurian species. Five additional species are also most likely capable of fission. The recent discovery of new fissiparous holothurian species indicates that this reproduction mode is more widespread in Holothuroidea than previously believed. New data about the history of the discovery of asexual reproduction in holothurians, features of fission, and regeneration of anterior and posterior fragments are described here. Asexual reproduction is obviously controlled by the integrated systems of the organism, primarily the nervous system. Special molecular mechanisms appear to determine the location where fission occurs along the anterior-posterior axis of the body. Alteration of the connective tissue strength of the body wall may play an important role during fission of holothurians. The basic mechanism of fission is the interaction of matrix metalloproteinases, their inhibitors, and enzymes forming cross-link complexes between fibrils of collagen. The population dynamics of fissiparous holothurians are discussed. PMID:25405228

  8. Virulence of lizard malaria: the evolutionary ecology of an ancient parasite-host association.

    Science.gov (United States)

    Schall, J J

    1990-01-01

    The negative consequences of parasitic infection (virulence) were examined for two lizard malaria parasite-host associations: Plasmodium agamae and P. giganteum, parasites of the rainbow lizard, Agama agama, in Sierra Leone, West Africa; and P. mexicanum in the western fence lizard, Sceloporus occidentalis, in northern California. These malaria species vary greatly in their reproductive characteristics: P. agamae produces only 8 merozoites per schizont, P. giganteum yields over 100, and P. mexicanum an intermediate number. All three parasites appear to have had an ancient association with their host. In fence lizards, infection with malaria is associated with increased numbers of immature erythrocytes, decreased haemoglobin levels, decreased maximal oxygen consumption, and decreased running stamina. Not affected were numbers of erythrocytes, resting metabolic rate, and sprint running speed which is supported by anaerobic means in lizards. Infected male fence lizards had smaller testes, stored less fat in preparation for winter dormancy, were more often socially submissive and, unexpectedly, were more extravagantly coloured on the ventral surface (a sexually dimorphic trait) than non-infected males. Females also stored less fat and produced smaller clutches of eggs, a directly observed reduction in fitness. Infected fence lizards do not develop behavioural fevers. P. mexicanum appears to have broad thermal buffering abilities and thermal tolerance; the parasite's population growth was unaffected by experimental alterations in the lizard's body temperature. The data are less complete for A. agama, but infected lizards suffered similar haematological and physiological effects. Infected animals may be socially submissive because they appear to gather less insect prey, possibly a result of being forced into inferior territories. Infection does not reduce clutch size in rainbow lizards, but may lengthen the time between clutches. These results are compared with

  9. Development of a novel multiplex PCR assay for rapid detection of virulence associated genes of Pasteurella multocida from pigs.

    Science.gov (United States)

    Rajkhowa, S

    2015-09-01

    As the pathogenicity of Pasteurella multocida is associated with various virulence factors (VFs), the aim of the study was to develop a novel multiplex PCR (m-PCR) assay for the rapid detection of important virulence associated genes (VAGs) of P. multocida isolates from pigs. The target recognized VFs used in the study were diverse adhesins (ptfA and pfhA), toxins (toxA), siderophores (tonB and hgbA), sialidases (nanB, nanH) and outer membrane proteins (ompA, ompH, oma87 and plpB). The primers for the genes encoding these VFs were designed by primer3 software (http://bioinfo.ut.ee/primer3-0.4.0/) using gene sequences available in Genbank. The detection limit of the developed assay was 10(2)  CFU ml(-1) . The m-PCR did not produce any nonspecific amplification products when tested against Bordetella bronchiseptica which also commonly infects pigs. We applied m-PCR to the field samples, and the results obtained were the same as the single PCR results. The developed assay would be very useful for veterinary diagnostic laboratories and for others interested in the rapid virulence profiling of porcine P. multocida isolates circulating in the piggeries. The study reports the development and evaluation of a novel multiplex PCR assay for the rapid detection of 11 important VAGs of Pasteurella multocida isolates from pigs. Rapid and simultaneous detection of recognized VFs of the organism are essential to know the virulo-types of P. multocida isolates circulating in the piggeries. The developed novel assay will be very useful for the rapid detection of VAGs of P. multocida isolates from pigs. © 2015 The Society for Applied Microbiology.

  10. Genomic characterization of Haemophilus parasuis SH0165, a highly virulent strain of serovar 5 prevalent in China.

    Directory of Open Access Journals (Sweden)

    Zhuofei Xu

    Full Text Available Haemophilus parasuis can be either a commensal bacterium of the porcine respiratory tract or an opportunistic pathogen causing Glässer's disease, a severe systemic disease that has led to significant economical losses in the pig industry worldwide. We determined the complete genomic sequence of H. parasuis SH0165, a highly virulent strain of serovar 5, which was isolated from a hog pen in North China. The single circular chromosome was 2,269,156 base pairs in length and contained 2,031 protein-coding genes. Together with the full spectrum of genes detected by the analysis of metabolic pathways, we confirmed that H. parasuis generates ATP via both fermentation and respiration, and possesses an intact TCA cycle for anabolism. In addition to possessing the complete pathway essential for the biosynthesis of heme, this pathogen was also found to be well-equipped with different iron acquisition systems, such as the TonB system and ABC-type transport complexes, to overcome iron limitation during infection and persistence. We identified a number of genes encoding potential virulence factors, such as type IV fimbriae and surface polysaccharides. Analysis of the genome confirmed that H. parasuis is naturally competent, as genes related to DNA uptake are present. A nine-mer DNA uptake signal sequence (ACAAGCGGT, identical to that found in Actinobacillus pleuropneumoniae and Mannheimia haemolytica, followed by similar downstream motifs, was identified in the SH0165 genome. Genomic and phylogenetic comparisons with other Pasteurellaceae species further indicated that H. parasuis was closely related to another swine pathogenic bacteria A. pleuropneumoniae. The comprehensive genetic analysis presented here provides a foundation for future research on the metabolism, natural competence and virulence of H. parasuis.

  11. Genomic Characterization of Haemophilus parasuis SH0165, a Highly Virulent Strain of Serovar 5 Prevalent in China

    Science.gov (United States)

    Zhou, Rui; Jin, Qi; Fan, Yang; Bei, Weicheng; Chen, Huanchun

    2011-01-01

    Haemophilus parasuis can be either a commensal bacterium of the porcine respiratory tract or an opportunistic pathogen causing Glässer's disease, a severe systemic disease that has led to significant economical losses in the pig industry worldwide. We determined the complete genomic sequence of H. parasuis SH0165, a highly virulent strain of serovar 5, which was isolated from a hog pen in North China. The single circular chromosome was 2,269,156 base pairs in length and contained 2,031 protein-coding genes. Together with the full spectrum of genes detected by the analysis of metabolic pathways, we confirmed that H. parasuis generates ATP via both fermentation and respiration, and possesses an intact TCA cycle for anabolism. In addition to possessing the complete pathway essential for the biosynthesis of heme, this pathogen was also found to be well-equipped with different iron acquisition systems, such as the TonB system and ABC-type transport complexes, to overcome iron limitation during infection and persistence. We identified a number of genes encoding potential virulence factors, such as type IV fimbriae and surface polysaccharides. Analysis of the genome confirmed that H. parasuis is naturally competent, as genes related to DNA uptake are present. A nine-mer DNA uptake signal sequence (ACAAGCGGT), identical to that found in Actinobacillus pleuropneumoniae and Mannheimia haemolytica, followed by similar downstream motifs, was identified in the SH0165 genome. Genomic and phylogenetic comparisons with other Pasteurellaceae species further indicated that H. parasuis was closely related to another swine pathogenic bacteria A. pleuropneumoniae. The comprehensive genetic analysis presented here provides a foundation for future research on the metabolism, natural competence and virulence of H. parasuis. PMID:21611187

  12. Preservation of enucleated porcine eyes for use in a wet laboratory

    NARCIS (Netherlands)

    Nibourg, Lisanne M.; Koopmans, Steven A.

    PURPOSE: To design a method to preserve enucleated porcine eyes for use in a wet laboratory. SETTING: Laboratory of Experimental Ophthalmology, University Medical Center Groningen, the Netherlands. DESIGN: Experimental study. METHODS: Porcine eyes were preserved using 15 methods including salt

  13. Virulence Plasmids of Spore-Forming Bacteria.

    Science.gov (United States)

    Adams, Vicki; Li, Jihong; Wisniewski, Jessica A; Uzal, Francisco A; Moore, Robert J; McClane, Bruce A; Rood, Julian I

    2014-12-01

    Plasmid-encoded virulence factors are important in the pathogenesis of diseases caused by spore-forming bacteria. Unlike many other bacteria, the most common virulence factors encoded by plasmids in Clostridium and Bacillus species are protein toxins. Clostridium perfringens causes several histotoxic and enterotoxin diseases in both humans and animals and produces a broad range of toxins, including many pore-forming toxins such as C. perfringens enterotoxin, epsilon-toxin, beta-toxin, and NetB. Genetic studies have led to the determination of the role of these toxins in disease pathogenesis. The genes for these toxins are generally carried on large conjugative plasmids that have common core replication, maintenance, and conjugation regions. There is considerable functional information available about the unique tcp conjugation locus carried by these plasmids, but less is known about plasmid maintenance. The latter is intriguing because many C. perfringens isolates stably maintain up to four different, but closely related, toxin plasmids. Toxin genes may also be plasmid-encoded in the neurotoxic clostridia. The tetanus toxin gene is located on a plasmid in Clostridium tetani, but the botulinum toxin genes may be chromosomal, plasmid-determined, or located on bacteriophages in Clostridium botulinum. In Bacillus anthracis it is well established that virulence is plasmid determined, with anthrax toxin genes located on pXO1 and capsule genes on a separate plasmid, pXO2. Orthologs of these plasmids are also found in other members of the Bacillus cereus group such as B. cereus and Bacillus thuringiensis. In B. thuringiensis these plasmids may carry genes encoding one or more insecticidal toxins.

  14. Metal acquisition and virulence in Brucella

    Science.gov (United States)

    Roop, R. Martin

    2013-01-01

    Similar to other bacteria, Brucella strains require several biologically essential metals for their survival in vitro and in vivo. Acquiring sufficient levels of some of these metals, particularly iron, manganese and zinc, is especially challenging in the mammalian host, where sequestration of these micronutrients is a well-documented component of both the innate and acquired immune responses. This review describes the Brucella metal transporters that have been shown to play critical roles in the virulence of these bacteria in experimental and natural hosts. PMID:22632611

  15. Microbial virulence and interactions with metals

    DEFF Research Database (Denmark)

    German, N.; Lüthje, F.; Hao, X

    2016-01-01

    Transition metals, such as iron, copper, zinc, and manganese play an important role in many bacterial biological processes that add to an overall evolutional fitness of bacteria. They are often involved in regulation of bacterial virulence as a mechanism of host invasion. However, the same transi...... reconstruction of Fe-S clusters and the use of Mn as a protectant against reactive oxygen species. Therefore, tight regulation of transition metal distribution in bacteria and hosts is a vital part of host-pathogen interactions....

  16. Identification of Secreted Exoproteome Fingerprints of Highly-Virulent and Non-Virulent Staphylococcus aureus Strains

    NARCIS (Netherlands)

    Bonar, Emilia; Wojcik, Iwona; Jankowska, Urszula; Kedracka-Krok, Sylwia; Bukowski, Michal; Polakowska, Klaudia; Lis, Marcin W; Kosecka-Strojek, Maja; Sabat, Artur J; Dubin, Grzegorz; Friedrich, Alexander W; Miedzobrodzki, Jacek; Dubin, Adam; Wladyka, Benedykt

    2016-01-01

    Staphylococcus aureus is a commensal inhabitant of skin and mucous membranes in nose vestibule but also an important opportunistic pathogen of humans and livestock. The extracellular proteome as a whole constitutes its major virulence determinant; however, the involvement of particular proteins is

  17. Reproductive endocrinology of vitamin D

    DEFF Research Database (Denmark)

    Lorenzen, Mette; Boisen, Ida Marie; Mortensen, Li Juel

    2017-01-01

    in the reproductive tissues. The reproductive organs are therefore responsive to and able to metabolize vitamin D locally. The exact role remains to be clarified but several studies have suggested a link between vitamin D and production/release of reproductive hormones into circulation, which will be the main focus...... suffering from reproductive problems and abnormal endocrinology research addressing the role of vitamin D in reproductive endocrinology is of clinical importance....

  18. Selenium toxicity and porcine focal symmetrical poliomyelomalacia: description of a field outbreak and experimental reproduction.

    OpenAIRE

    Wilson, T.M.; SCHOLZ, R. W.; Drake, T R

    1983-01-01

    An acute afebrile paretic condition was diagnosed in 18 of 225 feeder pigs between eight to ten weeks of age. Nine pigs died acutely, seven pigs were euthanatized and two appeared to recover. Macroscopic lesions in the ventral horns of the cervical and lumbar/sacral spinal cord enlargements consisted of focal, bilateral, depressed areas. Histopathologically, the lesion consisted of endothelial proliferation, glial cell reaction and microcavitation. Similar lesions were observed in some brain ...

  19. Selenium toxicity and porcine focal symmetrical poliomyelomalacia: description of a field outbreak and experimental reproduction.

    Science.gov (United States)

    Wilson, T M; Scholz, R W; Drake, T R

    1983-10-01

    An acute afebrile paretic condition was diagnosed in 18 of 225 feeder pigs between eight to ten weeks of age. Nine pigs died acutely, seven pigs were euthanatized and two appeared to recover. Macroscopic lesions in the ventral horns of the cervical and lumbar/sacral spinal cord enlargements consisted of focal, bilateral, depressed areas. Histopathologically, the lesion consisted of endothelial proliferation, glial cell reaction and microcavitation. Similar lesions were observed in some brain stem motor nuclei. High selenium levels were detected in the pig feed and in pig tissues and blood. Two of five experimental pigs fed a commercial grower ration and supplemented with 52 ppm selenium as sodium selenite developed paresis and paralysis after a 29 day feeding trial. Histopathological lesions of focal symmetrical poliomyelomalacia confined to the cervical and lumbar/sacral spinal cord enlargements, and identical to those in the field cases, were produced. Select brain stem motor nuclei were also affected.

  20. Heme oxygenase-1 metabolite biliverdin, not iron, inhibits porcine reproductive and respiratory syndrome virus replication.

    Science.gov (United States)

    Zhang, Angke; Duan, Hong; Li, Na; Zhao, Lijuan; Pu, Fengxing; Huang, Baicheng; Wu, Chunyan; Nan, Yuchen; Du, Taofeng; Mu, Yang; Zhao, Qin; Sun, Yani; Zhang, Gaiping; Hiscox, Julian A; Zhou, En-Min; Xiao, Shuqi

    2017-01-01

    Porcinereproductiveandrespiratorysyndromevirus (PRRSV) causes significant economic losses to the pork industry worldwide. Previously, we demonstrated that heme oxygenase-1 (HO-1) interferes with PRRSV replication. To elucidate the mechanisms involved, here we assess whether the HO-1 downstream metabolites biliverdin (BV) and/or iron mediate the HO-1 antiviral effect. We demonstrate a BV concentration-dependent suppression of PRRSV replication and show that virions are not directly inactivated by BV. Additionally, BV or N-acetyl cysteine (NAC) significantly reduced reactive oxygen species (ROS) in PRRSV-infected MARC-145 cells; however, because NAC did not reduce viral load, the BV antiviral effect is independent of decreased ROS levels. Moreover, a secondary metabolite of BV, bilirubin (BR), specifically mediates this anti-PRRSV activity via a nitric oxide (NO)-dependent cGMP/PKG signaling pathway. While increased iron via addition of FeCl3 did not interfere with PRRSV replication, iron depletion by deferoxamine (DFO) after cobalt-protoporphyrin IX induction of HO-1 did not restore PRRSV replication. Collectively, our findings identify a HO-1-BV/BR-NO-cGMP/PKG cascade as a novel pathway underlying the host cell antiviral effect. These results provide a unique insight into the molecular mechanisms underlying the antiviral effects of the stress-responsive protein HO-1 during PRRSV infection. Copyright © 2016 Elsevier Inc. All rights reserved.