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Wu, Ying, and 武盈. "Discovery and characterization of a novel porcine paramyxovirus." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hdl.handle.net/10722/196086.
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Most emerging infectious diseases in humans are zoonotic agents. Since the emergence of severe acute respiratory syndrome (SARS), swine-origin influenza and avian influenza epidemics, the study of novel and emerging viruses with zoonotic potential has been considered more and more important. Paramyxoviruses have been known for their potential to cross species barrier and infect new hosts. In the last decade, a number of novel and emerging paramyxoviruses have been reported in various animals. Our research group recently identified three novel bat paramyxoviruses, Tuhoko viruses 1, 2 and 3 (ThkPV-1, 2, and 3) from fruit bats in mainland China, an unclassified paramyxovirus, named Tailam virus (TlmPV) from Sikkim rats and a novel feline paramyxovirus, called Feline morbillivirus(FmoPV) from domestic cats in Hong Kong, suggesting that there is still a diversity of undescribed paramyxoviruses in animals.
In this study, a novel porcine paramyxovirus, Swine parainfluenza virus 1 (SpiPV-1), was discovered and characterized from deceased pigs in Hong Kong. A total of 951 samples from 386 deceased pigs were collected, including 386 nasopharyngeal swab, 303 rectal swab, 153 blood, 56 lung and 53 liver samples. And SpiPV-1 was detected in 12 (3.1%) of 386 nasopharyngeal swab and 2 (0.7%) of 303 rectal swab samples by RT-PCR. All the blood, lung and liver samples showed negative results. The complete genome sequences of three strains (SpiPV-1 S033N, SpiPV-1 S119N and SpiPV-1 S206N) from three pigs were amplified and determined. The genome organization of SpiPV-1 is similar to that of viruses under genus Respirovirus, subfamily Paramyxovirinae. The genome contains six genes (3’-N-P/V/C-M-F-HN-L-5’) and putatively codes for the nucleocapsid (N), phosphoprotein (P/V/C), matrix (M), fusion (F), attachment (HN) and large (L) proteins.Like other respiroviruses, the P gene of SpiPV-1 can produce more than one protein, including P, V and W proteins by mRNA editing and C protein by alternative translation initiation. And phylogenetic analysis showed that in all six phylogenetic trees constructed byusing the N, P, M, F, HN and L genes, the three strains SpiPV-1 S033N, S119N and S206N formed a distinct cluster among the known respiroviruses and were most closely related to Sendai virus (SenPV) and Human parainfluenza virus 1 (HpiPV-1). The genome organization, P gene analysis and phylogenetic analysis all suggested that SpiPV-1 is a novel paramyxovirus under genus Respirovirus, subfamily Paramyxovirinae. Seven porcine samples positive for SpiPV-1 were cultured in five different cell lines for viral isolation. However, no cytopathic effect was observed and no viral replication was detected in any of the cell lines.
The pathogenicity and emergent potential of SpiPV-1 remain to be determined. Further studies on serology and development of cell cultures for viral isolation may provide better insight into this novel paramyxovirus.published_or_final_version
Microbiology
Master
Master of Philosophy
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Bamford, Anona Isabelle. "Interactions between cytotoxic effector cells and bovine parainfluenza type 3 virus." Thesis, Queen's University Belfast, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.241326.
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Chan, Yuk-on. "Impact of respiratory viruses on mortality." Click to view the E-thesis via HKUTO, 2005. http://sunzi.lib.hku.hk/hkuto/record/b39724025.
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Norsted, Hanna. "The effect of interferon on the transcription pattern of parainfluenza virus 5." Thesis, University of St Andrews, 2013. http://hdl.handle.net/10023/3403.
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Interferon (IFN) is activated in response to virus infections and upregulates interferon-stimulated genes (ISGs) resulting in the expression of hundreds of proteins, many of which have direct or indirect antiviral activity. Parainfluenza virus 5 (PIV5) of the Paramyxoviridae family is a non-segmented negative sense single-stranded RNA virus with seven genes encoding eight proteins. Here we present that IFN induces alterations in the pattern of both virus transcription and translation and that ISG56 is primarily responsible for these effects. We report that when cells were treated with IFN post-infection, virus protein synthesis was inhibited while virus transcription levels were increased. These results suggest that ISG56 selectively inhibits the translation of viral mRNAs. In addition, the relationship of various PIV5 isolates was analysed by next generation sequencing. Four areas with a high degree of single nucleotide polymorphisms (SNPs) were identified and mapped to the intergenic regions of NP-V/P, M-F and HN-L, as well as the entire SH gene. Three of the isolates, the porcine strain SER and the canine strains CPI+ and CPI-, did not express an SH protein due to the lack of a start codon. A low degree of variation was found in the amino acid sequence of the HN glycoprotein suggesting that PIV5 may be less pressured to evolve in order to evade immune responses, such as neutralising antibodies.
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Chan, Yuk-on, and 陳旭安. "Impact of respiratory viruses on mortality." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2005. http://hub.hku.hk/bib/B39724025.
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Alias, Nadiawati. "Multivalent sialic acid binding proteins as novel therapeutics for influenza and parainfluenza infection." Thesis, University of St Andrews, 2014. http://hdl.handle.net/10023/4479.
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In nature, proteins with weak binding affinity often use a multivalency approach to enhance protein affinity via an avidity effect. Interested in this multivalency approach, we have isolated a carbohydrate binding module (CBM) that recognises sialic acid (known as a CBM40 domain) from both Vibrio cholerae (Vc) and Streptococcus pneumoniae (Sp) NanA sialidases, and generated multivalent polypeptides from them using molecular biology. Multivalent CBM40 constructs were designed either using a tandem repeat approach to produce trimeric or tetrameric forms that we call Vc3CBM and Vc4CBM, respectively, or through the addition of a trimerization domain derived from Pseudomonas aeruginosa pseudaminidase to produce three trimeric forms of proteins known as Vc-CBMTD (WT), Vc-CBMTD (Mutant) and Sp-CBMTD). Due to the position and flexibility of the linker between the trimerization domain and the CBM40 domain, site directed mutagenesis was employed to introduce a disulphide bond between the monomers at positions S164C and T83C of the CBM40 domain in order to promote a stable orientation of the binding site for easier access of sialic acids. Data from isothermal titration calorimetry (ITC) reveals that interaction of multivalent CBM40 proteins with α(2,3)-sialyllactose was mainly enthalpy driven with entropy contributing unfavorably to the interaction suggesting that these proteins establish a strong binding affinity to their ligand minimizing dissociation to produce stable multivalent molecules. However, using surface plasmon resonance (SPR), a mixed balance of entropy and enthalpy contributions was found with all constructs as determined by Van't Hoff plots. This proved that binding does not occur through a simple protein-ligand interaction but through disruption of hydrophobic and/or ionic hydration that provide the driving force to the process. Interestingly, the valency of multiple-linked polypeptides also plays an important part in the protein stabilization. However, little is known about their detailed structure when in multivalent form, as attempts to crystallize the whole protein molecule of Vc-CBMTD (WT) failed due to linker and domain flexibility. Only the trimerization domain (TD) part from Pseudomonas aeruginosa pseudaminidase was successfully crystallized and structure was determined to 3.0 Å without its CBM40 domain attached. In this thesis, we have also reported on the potential anti-influenza and anti- parainfluenza properties of these proteins, which were found to block attachment and inhibit infection of several influenza A and parainfluenza virus strains in vitro. As widely mentioned in literature, terminal sialic acids on the cell surface of mammalian host tissue provide a target for various pathogenic organisms to bind. Levels of viral inhibition were greatest against A/Udorn/72 H3N2 virus for Vc4CBM and Vc3CBM constructs with the lowest EC50 of 0.59 µM and 0.94 µM respectively, however most of the multivalent proteins tested were also effective against A/WSN/33 H1N1 and A/PR8/34 H1N1 subtypes. For parainfluenza virus, all constructs containing V. cholerae sialidase CBM40 domain showed great effect in inhibiting virus infection during cell protection assay. The best EC50 values were 0.2 µM from Vc-CBMTD (WT) followed by 1.17 µM from Vc4CBM and 1.78 µM from Vc-CBMTD (Mutant) which was against hPIV2, hPIV3 and hPIV5 infections respectively. Only a construct from S. pneumoniae sialidase known as Sp-CBMTD showed negligible effect on cell protection. All constructs were further tested for cytotoxicity in mammalian cell culture as well as undergoing an inhibition study on viral replication proteins. For the in vivo study, we also demonstrated the effectiveness of Vc4CBM to protect cotton rats and mice from hPIV3 and Streptococcus pneumoniae infections, when given intranasally in advance or on the day of infection. Therefore, these novel multivalent proteins could be promising candidates as broad-spectrum inhibitors or as a prophylactic treatment for both influenza and parainfluenza associated diseases.
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Baumgärtner, Wolfgang K. "Mechanisms of in vitro persistence of two canine paramyxoviruses and in vivo neuropathogenecity of canine parainfluenza virus /." The Ohio State University, 1986. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487265555440015.
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Roth, Jason Peter. "The Use of Reverse Genetics to Clone and Rescue Infectious, Recombinant Human Parainfluenza Type 3 Viruses." DigitalCommons@USU, 2009. https://digitalcommons.usu.edu/etd/467.
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Reverse genetics is a discipline that involves the use of genetic manipulation and modification to study an organism's altered phenotype. In this study, infectious recombinant viruses were rescued from altered cDNA clones encoding the antigenome of human parainfluenza virus type 3 and the resulting phenotypes were examined. In one clone, the gene for the enhanced green fluorescent protein was inserted into the virus antigenome to be expressed during viral replication, resulting in infected cells emitting green fluorescence. Viral titers, mRNA replication, and genomic replication for the virus expressing the enhanced green fluorescent protein were reduced when compared to the human parainfluenza virus type 3 wild-type strain. In addition, the sensitivity of the virus expressing the enhanced green fluorescent protein to antiviral compounds is increased when compared to the wild-type strain, which may lead to the identification of false positive antiviral compounds. An assay that measures the enhanced green fluorescent protein as a direct indicator of virus replication can be shortened to 3 days in duration and is a more robust assay compared to assays that measure cellular viability. In other clones, mutations were introduced into the phosphoprotein gene to eliminate the expression of the D domain of the PD protein in order to understand its function. The titers of two recombinant knockout viruses that are deficient in the expression of the D domain are reduced when compared to the wild-type strain in both MA-104 and A549 cells. In MA 104 cells, viral mRNA transcription and genomic replication of the two knockout viruses are reduced when compared to the wild-type strain. In A549 cells, cellular expression and secretion of antiviral cytokines infected with the two knockout viruses are either reduced or remain unchanged when compared to the wild-type strain. These results suggest that the D domain may play a role in viral RNA synthesis and not in counteracting the host cell's antiviral response. The results of these studies shed light on the influence an additional gene has on viral replication and possible functions of the D domain.
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Lam, Siu-yan. "Multiplex reverse transcription-PCR for detection and identification of human parainfluenza viruses 1,2,3 and 4 infection in hospitalized children with respiratory disease in Hong Kong /." View the Table of Contents & Abstract, 2007. http://sunzi.lib.hku.hk/hkuto/record/B3848058X.
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Xiao, Han. "Virus and interferon : a fight for supremacy : comparison of the mechanisms of influenza A viruses and parainfluenza virus 5 in combatting a pre-existing IFN-induced antiviral state." Thesis, University of St Andrews, 2011. http://hdl.handle.net/10023/2070.
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The Interferon (IFN) family of cytokines are produced in direct response to virus infection and they constitute the first line of defence against virus infection by inducing hundreds of interferon stimulated genes (ISGs) which act in concert to establish the so-called “antiviral state”. Influenza A viruses and parainfluenza virus type 5 (PIV5) are both small negative strand RNA viruses that must circumvent their hosts’ interferon (IFN) response for replication. However, the ways in which these viruses interact with the IFN system are very different. Although PIV5 replication is initially severely impaired in cells in a pre-existing IFN-induced antiviral state, it manages to overcome the antiviral state by targeting an essential component of type I IFN signalling, STAT1, for degradation. Thus the cells cannot maintain the antiviral state indefinitely without continuous signalling. Consequently, the virus resumes its normal replication pattern after 24-48 hours post-infection. In clear contrast, influenza virus fails to establish its replication in the majority of infected cells (90-95%) with a pre-existing IFN-induced antiviral state, although a few cells are still able to produce viral antigens. To further investigate how influenza virus interacts with cells in a pre-existing IFN-induced antiviral state, I have used in situ hybridization to follow the fate of input and progeny genomes in cells that have, or have not, been treated with IFN prior to infection. Here I show for the first time that IFN pre-treatment blocks the nuclear import of influenza A virus genome, which prevents the establishment of virus replication, but this can be overcome by increasing multiplicities of infection. Of those IFN-induced antiviral molecules, human MxA is an essential component of the IFN-induced antiviral state in blocking influenza virus genome import, as this block can be abolished by lentivirus-mediated knockdown of MxA. I also show that in cells constitutively expressing MxA the viral genome still manages to be transported into the nucleus, indicating that MxA might require an unidentified IFN-induced factor to block nuclear import of the influenza virus genome. These results reveal that IFN exerts its action at an early stage of virus infection by inducing MxA to interfere with the transport of viral genome into the nucleus, which is the factory for viral RNA production.
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Beneli, Patrícia Costa. "Associação entre fatores meteorológicos, poluentes atmosféricos e ocorrência de viroses respiratórias em crianças: destaque ao Parainfluenza Vírus Humano (HPIV)." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/87/87131/tde-02032011-181725/.
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As infecções respiratórias agudas contribuem para elevada morbimortalidade na infância, destacando o Parainfluenza (HPIV) nos quadros de crupe viral. Pouco é conhecido a influência dos fatores ambientais (meteorológicos e de poluição atmosférica) nas infecções respiratórias. De 21/10/2004 a 01/06/2007 foi conduzido um estudo ecológico de séries temporais, em menores de 15 anos, com sintomas respiratórios atendidos na Santa Casa de São Paulo e no Hospital Universitário de Jundiaí para determinar a freqüência de HPIV, pela imunofluorescência indireta e verificar a relação entre poluentes atmosféricos, variáveis meteorológicas na infecção respiratória. Os dados meteorológicos e de poluição ambiental foram coletados diariamente. Das 1464 amostras o HPIV foi detectado em 49 (5,5%) amostras (SCSP) e em 29(5,0%) amostras (HUFMJ),sendo o HPIV3 mais prevalente.O Ozônio e Dióxido de Nitrogênio tiveram relação com vírus respiratórios em São Paulo. Em Jundiaí observou-se relação com material particulado (lag3) e HPIV.Acute respiratory infections contribute to high infant morbidity and mortality, highlighting the parainfluenza (HPIV) in the frames of viral croup. Little is known the influence of environmental factors (meteorological and air pollution) in respiratory infections. From 21/10/2004 to 01/06/2007 was an ecological study of time series, for children under 15 years, with respiratory symptoms treated at Santa Casa de São Paulo and the University Hospital of Jundiaí to determine the frequency of HPIV by immunofluorescence and verify the relationship between air pollutants, meteorological variables in respiratory infection. The meteorological and environmental pollution were collected daily. 1464 samples of the HPIV was detected in 49 (5.5%) samples (SCSP) and 29 (5.0%) samples (HUFMJ) and the HPIV3 more prevalente.O Ozone and nitrogen dioxide were associated with respiratory viruses in Sao Paulo. In Jundiaí was observed relationship with particulate matter (lag3) and HPIV.
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Lam, Siu-yan, and 林小欣. "Multiplex reverse transcription-PCR for detection and identification of human parainfluenza viruses 1,2,3 and 4 infection in hospitalizedchildren with respiratory disease in Hong Kong." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2007. http://hub.hku.hk/bib/B45011357.
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Amaral, Larissa Morais Bomilcar do. "Epidemiologia e caracterização molecular do Vírus Parainfluenza Humano 1, 2 e 3 em crianças menores de 5 anos de idade atendidas no Hospital Universitário em 2007, São Paulo - Brasil." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/42/42132/tde-02022010-093628/.
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O objetivo do presente trabalho é descrever o perfil epidemiológico e molecular dos vírus Parainfluenza em crianças menores de 5 ano de idade, com infecção das vias respiratórias. Por tanto, aspirados de nasofaringe de 742 crianças foram examinadas para os Vírus Parainfluenza Humano(HPIV1, HPIV2 e HPIV3), hRSV, hMPV e IA e IB pela técnica de RT-PCR, durante o ano de 2007. Ao todo foram identificados 52(7%) Parainfluenza vírus, sendo 9(17,3%) HPIV1, 8(15,4%) HPIV2 e 35(67,3%) HPIV3. Destas, 12 (23%) foram casos de coinfecções, sendo 3 (25%) de HPIV3 com VRS, 3 (25%) com MPV e 3 (25%) com HPIV1 e 1(8,33%) com HPIV2; além de 1(8,33%) do HPIV1 com hVSR; e 1(8,33%) de HPIV2 com o hMPV. Com relação ao setor de atendimento, dos 52 pacientes com HPIV, 8(5%) dos casos foram atendidos no PA; 38(8,4%) na ENF; 6(18,2%) na UTI. Obtivemos, no estudo, 21(5,3%) crianças do sexo masculino com infecção por HPIV e 31(9%) de sexo feminino. Quanto a idade, as crianças entre 1 a 23 meses foram as mais frequentemente infectadas. Com relação ao diagnóstico clínico tivemos 19(36,5%) casos com bronquiolites, 13(25%) casos com pneumonia, 7(13,4%) com broncopneumonia, 2(3,9%) de casos de dispnéia e 2(3,9%) de IVAS, além de 9(17,3%) sem informação da sintomatologia. A sazonalidade do HPIV foi marcada pela circulação do HPIV3 durante o ano inteiro, com maior incidência em outubro(primavera). O HPIV 1 e 2 se alternaram durante o ano, onde o HPIV1 circulou no primeiro semestre com maior incidência no mês de abril e maio e o HPIV2 circulou no segundo semestre com maior incidência em outubro, coincidindo com o HPIV3. O Estudo filogenético dos HPIV1 demonstrou 9 mutações nucleotídicas, sendo que 4 são características das amostras brasileiras. Entretanto, quando feito a transdução para aminoácido, apenas uma mutação foi não silenciosa, onde observamos a mudança de glicina, nas amostras do Genbank, para argenina, nas amostras brasileiras, deixando-as em um clado único na topologia da árvore obtida. O HPIV3 apresentou 4 subgrupos distintos durante o ano e suas mutações tanto nucleotídicas quanto de aminoácidos foram todas silenciosas. Em conclusão o vírus parainfluenza representaram a 3ª maior causa de infecção das vias respiratórias inferiores, precedido pelo vírus respiratório Sincicial e o Metapneumovirus.The goal of the present work was to characterize the epidemiologic and molecular profile of the parainfluenza virus in children with less than 5 years of age and presenting respiratory tract infection. Human Parainfluenza (HPIV1, HPIV2, HPIV3), and hRSV, hMPV e IA e IB were evaluated in nasopharyngeal aspirate from 742 children by RT-PCR during the year of 2007. Human parainfluenza was identified in 52(7%) of the samples, which include 9(17,3%) HPIV1, 8(15,4%) HPIV2 and 35(67,3%) HPIV3 17,3% (n=9). Among the 52 cases with parainfluenza, 12 (23%) presented coinfection with other viruses: 3 (25%) coinfection with HPIV3 and VRS, 3 (25%) MPV , 3 (25%) HPIV1 , 1(8,33%) HPIV2 . In addition, it was observed 1(8,33%) of cases with HPIV1 and hVSR; and 1(8,33%) of cases with HPIV2 and hMPV. With respect to the clinical care of the 52 patients with HPIV, 8(5%) were treated at the ER? ; 38(8,4%) at the ENF and 6(18,2%) at the ICU. With respect to gender, a total of 21(5,3%) of the participants with HPIV infections were male and 31(9%) were female. The prevalent age range of participants with infections was 1-23 months of age. The following clinical parameters were observed: bronchiolitis 19(36,5%), pneumonia 13(25%), bronchopneumonia 7(13,4%), dyspnea 2(3,9%) and IVAS 2(3,9%). No clinical information was available for 9(17,3%) cases. HPIV3 infections were detected all year long with an incidence peak in October (Spring). HPIV1 and 2 infections were intercalated during the year. HPIV1 was detected during the first semester with peaks of incidence in April and May whereas HPIV2 was detected during the second semester with peaks of incidence in October, coexisting with HPIV3. HPIV1phylogenetic studies revealed 9 genetic mutations. 4 out of the 9 mutations are exclusively found in the brazilian population. However, only one of these 9 mutations is non-silent and causes a glycine to argenine substitution. When compared to other brazilian sequences in the GenBank, this observation revealed um clado único na topologia da árvore obtida. HPIV3 genotyping included 4 distinct groups detected all year long. All mutations observed in HPIV3 were silent. In conclusion, the parainfluenza virus represent the 3rd major cause of infection of the lower respiratory tract, following the sincicial respiratory virus and the Metapneumovirus.
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Goka, Edward Anthony Chilongo. "Influenza A viruses dual and multiple infections with other respiratory viruses and risk of hospitalization and mortality." Thesis, University of Manchester, 2014. https://www.research.manchester.ac.uk/portal/en/theses/influenza-a-viruses-dual-and-multiple-infections-with-other-respiratory-viruses-and-risk-of-hospitalization-and-mortality(256eb122-a52a-4276-8dc1-28b5a2cc6662).html.
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Introduction: Epidemiological studies have indicated that 5-38% of influenza like illnesses (ILI) develop into severe disease due to, among others, factors such as; underlying chronic diseases, age, pregnancy, and viral mutations. There are suggestions that dual or multiple virus infections may affect disease severity. This study investigated the association between co-infection between influenza A viruses and other respiratory viruses and disease severity. Methodology: Datum for samples from North West England tested between January 2007 and June 2012 was analysed for patterns of co-infection between influenza A viruses and ten respiratory viruses. Risk of hospitalization to a general ward ICU or death in single versus mixed infections was assessed using multiple logistic regression models. Results: One or more viruses were identified in 37.8% (11,715/30,975) of samples, of which 10.4% (1,214) were mixed infections and 89.6% (10,501) were single infections. Among patients with influenza A(H1N1)pdm09, co-infections occurred in 4.7% (137⁄2,879) vs. 6.5% (59⁄902) in those with seasonal influenza A virus infection. In general, patients with mixed respiratory virus infections had a higher risk of admission to a general ward (OR: 1.43, 95% CI: 1.2 – 1.7, p = <0.0001) than those with a single infection. Co-infection between seasonal influenza A viruses and influenza B virus was associated with a significant increase in the risk of admission to ICU/ death (OR: 22.0, 95% CI: 2.21 – 219.8 p = 0.008). RSV/seasonal influenza A viruses co-infection also associated with increased risk but this was not statistically significant. For the pandemic influenza A(H1N1)pdm09 virus, RSV and AdV co-infection increased risk of hospitalization to a general ward, whereas Flu B increased risk of admission to ICU/ death, but none of these were statistically significant. Considering only single infections, RSV and hPIV1-3 increased risk of admission to a general ward (OR: 1.49, 95% CI: 1.28 – 1.73, p = <0.0001 and OR: 1.34, 95% CI: 1.003 – 1.8, p = 0.05) and admission to ICU/ death (OR: 1.5, 95% CI: 1.20 – 2.0, p = <0.0001 and OR: 1.60, 95% CI: 1.02 – 2.40, p = 0.04). Conclusion: Co-infection is a significant predictor of disease outcome; there is insufficient public health data on this subject as not all samples sent for investigation of respiratory virus infection are tested for all respiratory viruses. Integration of testing for respiratory viruses’ co-infections into routine clinical practice and R&D on integrated drugs and vaccines for influenza A&B, RSV, and AdV, and development of multi-target diagnostic tests is encouraged.
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Schomacker, Henrick. "Rekombinante bovin-humane Parainfluenzaviren Typ 3 als Impfvektoren gegen nicht-virale Antigene." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2008. http://dx.doi.org/10.18452/15785.
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Bei bhPIV3 handelt es sich um ein bovines Parainfluenzavirus Typ 3 (bPIV3), dessen Ober-flächenproteingene gegen jene des humanen Parainfluenzavirus Typ 3 (hPIV3) ausgetauscht wurden. Dieses ursprünglich als experimenteller Impfstoff gegen hPIV3 entwickelte Virus wurde darüber hinaus als Impfvektor zur Expression anderer viraler Antigene verwendet. Im Rahmen der hier vorgestellten Arbeit wurden die ersten bhPIV3-basierten Vektoren für nicht-virale Antigene hergestellt und in einem ersten Versuch evaluiert. Dazu wurden ein reverses Genetiksystem zur Herstellung rekombinanter bhPIV3 in einem neuen Labor aufgebaut und fünf neue rekombinante Viren erhalten, welche zusätzlich Antigene des Mycobacterium tuberculosis (M. tb.) exprimieren. Balb/c-Mäuse wurden intranasal mit den bhPIV3-Vektoren infiziert, so dass sowohl deren Replikation als auch der induzierte protektive Effekt gegenüber M. tb.-Neuinfektionen getestet werden konnte. In einem ersten Versuch zeigte sich, dass eine Immunisierung mit den rekombinanten Viren allein keine Schutzwirkung entfaltet. Als Boost-Impfung nach Gabe des Bacille Calmette Guérin (BCG) zeigten einige Vektoren jedoch einen signifikanten protektiven Effekt. In einem Folgeversuch konnten diese Beobachtungen jedoch bislang nicht bestätigt werden, so dass weitere Versuche durchzuführen sind, bevor eine endgültige Aussage bezüglich des hervorgerufenen Schutzeffektes getroffen werden kann. In einem weiteren Tierversuch wurde gezeigt, dass die Baumwollratte ein Tiermodell darstellt, in dem bhPIV3 erheblich schlechter repliziert als hPIV3. Trotz der eingeschränkten Replikation induzierte bhPIV3 neutralisierende Antikörpertiter gegen hPIV3, die mit durch hPIV3 induzierten Titern vergleichbar waren. Mit Hilfe eines neu generierten rekombinanten Virus, welches das grün fluoreszierende Protein EGFP exprimiert, konnte ein Weg aufgewiesen werden, die Bestimmung neutralisierender Antikörpertiter deutlich zu vereinfachen.The initial objective of this project was to establish a reverse genetic system for generation of recombinant bovine/human parainfluenza virus type 3 (bhPIV3), a bovine PIV3 (bPIV3) in which the bhPIV3 glycoprotein genes are replaced by their counterparts of human PIV3 (hPIV3). In addition, methods needed to characterise virus infectivity, genetic integrity and relevant in vitro phenotypes were established. The reverse genetics system was used to add individual mycobacterium tuberculosis (M. tb.) open reading frames (ORFs) as supernumerary gene units to the bhPIV3 genome and to rescue bhPIV3 vectors that expressed M. tb. antigens. In addition, a similar vector expressing the enhanced green fluorescent protein (EGFP) was constructed. Following the in vitro characterization of the derived viral vectors, the M. tb. vectors were evaluated for their efficacy to protect against M. tb. aerosole challenge in the Balb/c mouse model for tuberculosis. Although, in a single experiment, vaccination with bhPIV3 vectors alone did not confer any protection against M. tb. challenge, a boost with selected bhPIV3 vectors after Bacille Calmette Guérin (BCG) priming was successful in conferring protective efficacy against M. tb. challenge. A repeat of this challenge study could not confirm the initial observation, and further experiments are needed to determine whether the observed protection can be reliably reproduced. Evaluation of the bhPIV3 vectors in the cotton rat model showed that this small animal model is suitable to evaluate the attenuation phenotype of bhPIV3 compared to human parainfluenza virus type 3 (hPIV3). Although replication of bhPIV3 was highly restricted compared to hPIV3, hPIV3 neutralizing antibody titers induced by bhPIV3 infection were similar to those induced by hPIV3 infection. Studies with bhPIV3 expressing EGFP led to a new fluorescence based assay to determine hPIV3 neutralizing antibody titers. This assay could save time and resources in hPIV3 serology.
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Saffran, Holly Anne. "Regulation of human parainfluenza virus type 3 transcription." Thesis, University of Ottawa (Canada), 1995. http://hdl.handle.net/10393/9503.
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To elucidate the roles of the junctional elements in HPIV3 transcription, cDNAs were constructed containing CAT and luciferase reporter genes flanked by sequences representing the HPIV3 termini necessary for transcription, replication and packaging. Mutations to the gene end sequence abolished expression of the upstream and downstream genes. Deleting the gene start sequence at the junction resulted increased expression of the upstream gene, but abrogated downstream gene activity. Alterations in the length of the intergenic trinucleotide resulted in decreased expression of both upstream and downstream genes. Mutations in the sequence of this nontranscribed trinucleotide resulted in decreased activity of the upstream gene but no change in expression of the downstream gene. The gene end sequence does not appear to contain the only signals for termination of transcription. The purine trinucleotide intergenic region is important for termination, but only the presence of three nucleotides appears to be necessary and sufficient for expression of the following gene. Results obtained from assaying reporter activity could often be interpreted in several ways. For example, the data could not distinguish between polymerase readthrough and premature termination. Two RNA detection methods were investigated and show promise as means for detecting and analyzing specific RNA species in transfected cells. (Abstract shortened by UMI.)
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Chapman, Amanda Ruth. "Regulation of the human parainfluenza virus (hPIV3) fusion protein." View the abstract Download the full-text PDF version, 2008. http://etd.utmem.edu/ABSTRACTS/2008-048-Chapman-index.htm.
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Thesis (M.S.)--University of Tennessee Health Science Center, 2008.Title from title page screen (viewed on January 6, 2009). Research advisor: Charles J. Russell, Ph.D. Document formatted into pages (ix, 41p. : ill.). Vita. Abstract. Includes bibliographical references (p. 38-41).
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Smielewska, Anna Alexandra. "Human parainfluenza virus 3 : genetic diversity, virulence and antiviral susceptibility." Thesis, University of Cambridge, 2019. https://www.repository.cam.ac.uk/handle/1810/287954.
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Human parainfluenza 3 (HPIV3) is a member of the Paramyxoviridae, a single strain negative-sense non-segmented RNA virus in the order Mononegavirales. It is a respiratory pathogen with a broad spectrum of presentations for which there is currently neither a vaccine nor licensed treatment for HPIV3. To date most research on HPIV3 has been conducted using significantly culture adapted reference strains. Therefore, minimally adapted clinical strains were grown in two cell culture systems: immortalised and primary. Plaque phenotype, growth kinetics and inflammatory response triggered were evaluated and it was found that there is a range of phenotypes exhibited by clinical strains with potential implications in vivo. To examine the genetic diversity of circulating strains of HPIV3 in the UK, a new amplicon based sequencing pipeline for whole genome sequencing of HPIV3 was developed and validated. A short hypervariable region in the HPIV3 genome was identified and evaluated as a potential candidate for subsequent phylogenetic analysis compared to whole genome data. This method was then applied to tracking an HPIV3 outbreak that took place on a paediatric oncology ward. It was found to be a point-source outbreak and the clinical impact in this setting, as well as the infection control procedures involved were evaluated. Finally a robust in vitro model for the evaluation of potential therapeutic candidates for HPIV3, based on a panel of minimally passaged clinical strains as well as a culture-adapted reference strain, was set up. This model was applied to three potential inhibitors of HPIV3: ribavirin, favipiravir and zanamivir. The results showed that clinical strains were at least as susceptible to ribavirin and favipiravir as the laboratory reference strain and significantly more susceptible to zanamivir. This indicates that further work on minimally adapted clinical strains is essential to further the understanding of this important virus.
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Murphy, Lise. "Regulation of human parainfluenza virus type 3 fusion protein expression." Thesis, University of Ottawa (Canada), 2004. http://hdl.handle.net/10393/26723.
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Human parainfluenza virus type 3 (HPIV3) is an enveloped, negative-strand, non-segmented RNA virus. HPIV3 is a human respiratory pathogen that primarily causes diseases such as croup, bronchiolitis and pneumonia in children. The virus has two glycoproteins that allow it to interact with host cells, the receptor binding protein hemagglutinin-neuraminidase (HN) and the fusion protein (F), which enables the virus to enter the cell by fusion of the viral envelope to the target cell plasma membrane.
This research was initiated with the goal of determining mechanisms by which HPIV3 regulates the expression of the fusion protein. Evidence indicated that the transcription of the F gene differed from that of the other genes because there was a very high level of read-through transcription at the junction of the matrix (M) gene and the F gene. This read-through transcription caused >80% of the potential F gene transcripts to be present in biscistronic M/F mRNA, in which the F open reading frame cannot be translated. (Abstract shortened by UMI.)
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Jogalekar, Prachi. "Analysis of gene junction sequences of human parainfluenza virus type 3." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0019/MQ58465.pdf.
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Chidgey, Sharon Michelle. "Airway function, inflammation and pulmonary histopathology following parainfluenza-3 virus infection." Thesis, Cardiff University, 2007. http://orca.cf.ac.uk/55663/.
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Human parainfluenza viruses (PIV) 1, 2, 3 and 4 (A and B) cause approximately 39-40% of all acute respiratory infections in infants and children for which there is no effective therapy. Firstly, this thesis was aimed at ascertaining a suitable guinea pig model of PIV-3 infection by determining the most efficient route of virus application. Secondly, to ascertain if a temporal association may be established during the time course of infection (Day 1-40) by measuring the following parameters: body weight, rectal temperature, airway function (sGaw), airways reactivity to inhaled histamine, inflammatory cell infiltration to the lungs measured by bronchoalveolar lavage, wet lung weights, inflammatory markers (nitric oxide and total protein levels), lung histological analysis and recovery of the virus from bronchoalveolar lavage fluid and lung tissue. Further to this, experiments were also designed to determine the impact of the glucocorticoid, dexsamethasone (in vivo and in vitro) and the phosphodiesterase inhibitor, rolipram (in vivo). Lastly, this study ascertained the effect of pre-sensitisation with antigen on the responses of antigen in PIV-3 infected guinea pigs. These investigations have shown guinea pigs to be a suitable model for PIV-3 infection and intranasal inoculation is the most efficient route of virus application. In addition, these investigations also established a temporal association between the time course of PIV-3 infection including airway reactivity to histamine, airway function (sGBW), airways reactivity to inhaled histamine, inflammatory cell infiltration, wet lung weights, inflammatory markers (nitric oxide and total protein levels), pulmonary histopathology and recovery of the virus from bronchoalveolar lavage fluid and lung tissue. Pre-treatment of PIV-3 infected guinea pigs with the glucocorticoid, dexamethasone and the phosphodiesterase inhibitor, rolipram (in vitro) ameliorated the inflammatory response and airway hyperreactivity. Unlike rolipram, dexamethasone also reduced viral titre (in vivo and in vitro), supporting a role for the anti-viral effects of dexamethasone. The anti-inflammatory effects of dexamethasone are well known and it has been speculated by other authors which maybe caused by decreased viral receptors on the epithelial cells via inhibition of intracellular adhesion molecule-1 expression. Finally, in this study PTV-3 infection, enhanced the effect of pre-allergen sensitisation, which may arise from increased permeability of the airway mucosa to allergens, due to damage of the respiratory epithelium and increased recruitment of dendritic cells. In summary, this thesis has established a clearly defined efficacy of dexamethasone use in the treatment of PIV-3 infection.
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Bradford, Hannah Elizabeth Linda. "Cell-mediated immunity to parainfluenza type 3 virus in young calves." Thesis, Queen's University Belfast, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.334497.
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Sieg, Scott F. "Infection and immunoregulation of T lymphocytes by parainfluenza virus type 3." Case Western Reserve University School of Graduate Studies / OhioLINK, 1996. http://rave.ohiolink.edu/etdc/view?acc_num=case1057593999.
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Short, John A. L. "Defective interfering particles of parainfluenza virus subtype 5 and interferon induction." Thesis, University of St Andrews, 2015. http://hdl.handle.net/10023/7036.
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The innate immune response is the first line of defence against virus infection. Cells contain a diverse array of pathogen recognition receptors (PRRs) that are able to recognise multiple pathogen associated molecular patterns (PAMPS) that present themselves during virus infection. The RIG-I (Retinoic acid inducible–gene-I) and MDA5 (melanoma differentiation- associated gene 5) PRRs detect specific viral RNA ligands and subsequently induce the expression of the cytokine Interferon-β(IFN-β). IFN-βis secreted, acting on the infected cell and neighbouring uninfected cells to generate an antiviral state that is hostile to virus transcription, replication and dissemination, whilst also orchestrating adaptive immune responses. Given IFN-βs crucial cellular antiviral role, understanding its induction is of great importance to developing future antiviral drugs and vaccine strategies. Using A549 reporter cells in which GFP expression is under the control of the IFN-βpromoter, we show that there is a heterocellular response to parainfluenza virus 5 (PIV5) and infection with other negative sense RNA viruses. Only a limited number of infected cells are responsible for IFN-βinduction. Using PIV5 as a model, this thesis addresses the nature of the PAMPs that are responsible for inducing IFN-βfollowing PIV5 infection. The previous work has shown that PIV5 Defective Interfering particle (DI) rich virus preparations acted as a better inducer of IFN-βcompared to DI poor stocks. DIs are incomplete virus genomes produced during wild-type virus replication as a result of errors in the viral polymerase. To investigate this further, A549 Naïve, MDA5/RIG-I/LGP2 Knock down reporter cells were infected with PIV5 W3 at a low MOI to examine the inverse correlation of NP and GFP of DIs generated during virus replication and not from the initial infection. GFP+ve cells were cell sorted, and using QPCR it was found that cells that have the IFN-βpromoter activated contain large amounts of DIs relative to GFP-ve cells. This data supports the Randall group's findings that DIs generated during errors of wild-type replication by the viral RNA polymerase are the primary PAMPs that induce of IFN-β, as opposed to PAMPs being generated during normal wild-type virus replication.
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Storey, Douglas Gordon. "Structural characterization of human parainfluenza virus 3 and cloning of viral specific genes." Thesis, University of Ottawa (Canada), 1987. http://hdl.handle.net/10393/5476.
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Murphy, Donald G. "Defective interfering particles of human parainfluenza virus 3 and establishment of persistent infections." Thesis, University of Ottawa (Canada), 1990. http://hdl.handle.net/10393/5615.
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Defective interfering particles of human parainfluenza virus 3 (HPIV3) were generated and/or amplified by serial undiluted passage of the standard virus. Analysis of the progeny virus titer at each cell passage revealed a cyclic pattern of virus production. Viruses produced from serial passages 8 and 9 interfered with replication of the standard virus. When cells were mixedly infected with standard virus and virus from either serial passages 5 or 8, three subgenomic RNA species, in addition to the standard virus genomic RNA, were detected in the progeny virions. Northern blot analysis revealed that all three subgenomic RNAs contained sequences from the 5$\sp\prime$-end of the standard virus genome. The data indicates that 5$\sp\prime$ defective interfering (DI) particles were generated and/or amplified during serial undiluted passage of HPIV3. Two independent HPIV3 persistent infections of LLC-MK$\sb2$ cells were established. One persistently infected culture was established by propagating the few cells which survived from an acute standard virus infection. The other persistently infected culture was readily established with virus from serial undiluted passage 8. This suggests that the DI particles present in serial passage 8 virus protected the cells from standard virus destruction. The two persistently infected cultures were propagated for nearly three years and the virus released from both cell lines at all times examined was noncytocidal. Subgenomic RNAs (putative DI particle genomes) were detected in virions released from both persistently infected cultures at various cell passages. No apparent changes in the size of the viral proteins were observed throughout the persistent infections. The DI particle-size genomes and the noncytocidal mutant virus are both likely to be involved in the maintenance of the persistent state. The degree of genetic change undergone by the viral genome during long-term persistence was examined. Nucleotide sequence analysis of the M gene and flanking regions of virus recovered after 147 cell passages (29 months of persistence) revealed that the viral genome did not undergo extensive genetic change. In the M protein coding region, only one conservative amino acid change was observed indicating that M protein mutation is not likely to be involved in the maintenance of the persistent infections. In contrast, the genomic 3$\sp\prime$-termini of viruses recovered after 146 cell passages was found to be highly mutated. The mutations observed were either U to C or A to G changes. Such exclusive substitution of one type of nucleotide for another is unlikely to be due to intrinsic polymerase errors but to an extrinsic biased hypermutational activity. An RNA unwinding/modifying activity that could give rise to the biased hypermutations is discussed.
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Cote, Marie-Jose. "The human parainfluenza virus 3 fusion protein: Cloning, mapping, sequence analysis and expression." Thesis, University of Ottawa (Canada), 1989. http://hdl.handle.net/10393/20781.
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Hartwagner, Nadja A. "Dissection of cell entry of Sendai virus and bovine parainfluenza 3 virus by electron microscopy at high resolution /." [S.l.] : [s.n.], 2009. http://opac.nebis.ch/cgi-bin/showAbstract.pl?sys=000286607.
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Ebata, Sharon Nori. "Requirements for syncytium formation mediated by the fusion glycoprotein of human parainfluenza virus type 3." Thesis, University of Ottawa (Canada), 1996. http://hdl.handle.net/10393/10423.
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Recombinant vaccinia viruses VF and VHN were shown to express glycoproteins with molecular weights similar to authentic HPIV3 F and HN proteins that were transported to the cell surface and recognized by monoclonal antibodies specific for HPIV3 F and HN. The HN glycoprotein in VHN-infected cells exhibited both hemagglutinin and neuraminidase activities. Both cleaved and uncleaved forms of the F glycoprotein were immunizers from VF-infected cell lysates. Fusogenic activity of the F protein was demonstrated only upon coinfection of HeLa T4$\sp+$ cells with VF + VHN and resulted in syncytium formation and hemolysis. Recombinant adenoviruses AdF and AdHN, expressed the HPIV3 F and HN proteins on the surface of infected HeLa T4$\sp+$ cells, respectively. AdHN-infected cells produced HN protein that was glycosylated and exhibited both hemagglutinin and neuraminidase activities. AdF infection led to the synthesis of glycosylated F$\sb0$ that was proteolytically cleaved. As was observed for vaccinia virus recombinants, syncytium formation was only observed upon coinfection of HeLa T4$\sp+$ cells with AdF + AdHN The F and HN proteins expressed by recombinant adenoviruses were recognized by monoclonal antibodies specific for the HPIV3 F or HN protein and were immunogenic in mice. Sera from AdF-or AdHN-inoculated mice immunoprecipitated appropriate glycoproteins from lysates of HPIV3-infected cells and neutralized HPIV3. Amino acid substitutions were introduced into three regions of the HPIV3 F protein to identify sequences involved in fusogenic activity. Substitution of uncharged amino acids for positively charged residues within the cytoplasmic tail, or introduction of a charged residue within the transmembrane domain, did not appear to affect fusogenic activity. Several different mutations designed to disrupt the leucine zipper or the heptad repeat motifs resulted in mutant F proteins that not only exhibited decreased fusogenic activity but concomitantly lost the ability to react with monoclonal antibody c128, which recognized an epitope in antigenic site B of the HPIV3 F glycoprotein. An association between the heptad repeat and leucine zipper domains, therefore, appears to be required for the HPIV3 F glycoprotein to adopt a conformation that is recognized by antibody c128 and that appears to be important for fusogenic activity.
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Vaucher, Rodrigo de Almeida. "Desenvolvimento de técnicas de RT-PCR para seqënciamento do gene da hemaglutinina-neuraminidase (HN) e detecção do vírus Parainfluenza bovino tipo 3." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2005. http://hdl.handle.net/10183/8932.
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Existem diversos trabalhos publicados sobre a utilização de diferentes métodos imunológicos para diagnosticar infecções do trato respiratório causadas por vírus parainfluenza bovino tipo 3 (bPIV-3). Entretanto, é escassa a literatura sobre a utilização da técnica de isolamento viral. Até o presente momento não havia sido relatada a utilização da Transcrição Reversa - Reação em Cadeia da Polimerase (RT-PCR), na detecção de bPIV-3. O objetivo deste estudo foi contribuir para uma melhor caracterização dos bPIV-3 através do desenvolvimento de técnicas de RTPCR para a sua detecção. Utilizando-se uma amostra referência de bPIV-3 (SF-4) e uma amostra de bPIV-3 isolada no Rio Grande do Sul (amostra DIO) foram desenvolvidas técnicas de RT-PCR para a amplificação de diferentes regiões do gene da hemaglutinina-neuraminidase (HN). Após seqüenciamento parcial do gene HN e alinhamento das seqüências da amostra DIO, os resultados revelaram homologia de 99% em relação à amostra referência, 98% a 91% quando comparada com outras amostras de bPIV-3 previamente publicadas na rede e 79% a 80% quando comparada com as amostras de hPIV-3. Foi desenvolvida, também, uma técnica de RT-PCR para amplificação de parte do gene da HN do bPIV-3 e do hPIV-3. Nos experimentos de otimização, a técnica de RT-PCR, comparada com o isolamento viral, apresentou sensibilidade de 140 DICC50, boa especificidade e reprodutibilidade. Os resultados, após seqüenciamento da amostra de vírus bPIV-3 isolada no RS, apresentaram, grande homologia com os das amostras de bPIV-3 comparadas, especialmente a amostra referência. Os dados obtidos neste estudo mostraram que a RT-PCR desenvolvida para a detecção de PIV-3 foi capaz de amplificar um fragmento de 1009 bp do gene da HN de amostras de PIV-3 isoladas de bovinos e humanos, possibilitando a sua utilização em diagnóstico e em estudos epidemiológicos.There are several published studies on the application of different immunological methods for diagnosis of respiratory infections caused by bovine type 3 Parainfluenza virus (bPIV-3). However, the literature on viral isolation procedure is very scarce. At present there is no report about the utilization of reverse transcription-polymerase chain reaction (RT-PCR) for bPIV-3 detection. The aim of this study was to contribute to a better caracterization of bovine type 3 Parainfluenza virus by developing RT-PCR techniques to its detection. A reference sample (SF-4) and a sample of bPIV-3 isolated in Rio Grande do Sul (DIO sample) were used to develop RT-PCR techniques by amplification of different regions of hemagglutinin-neuraminidase gene (HN). After HN gene partial sequencing of DIO sample and sequence alignment, the results revealed 99% homology when compared to reference sample, 98% the 91% homology in relation to bPIV-3 samples previously published and 79% the 80% if compared to hPIV-3 samples. It was also developed a RT-PCR for amplification of a part of bPIV-3 and hPIV-3 NH gene. In optimization experiments, compared to viral isolation procedure, the RTPCR displayed a sensitivity of 140 DICC50, good specificity and reproductibility. Results after sequencing of bPIV-3 sample isolated in RS displayed strong homology with those of bPIV-3 tested samples, specially the reference sample. Data obtained in this study showed that the RT-PCR technique developed for PIV- 3 detection was able to amplify an 1009 bp HN gene fragment of bovine and human PIV-3 samples which enables its utilization in diagnostic and epidemiological studies.
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Terrier, Olivier. "Les glycoprotéines d'enveloppe des virus Parainfluenza : étude structure versus fonctions et développement d'applications diagnostiques et thérapeutiques." Lyon 1, 2009. http://www.theses.fr/2009LYO10011.
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Les virus parainfluenza humains (hPIV, Famille des Paramyxoviridae) sont des virus respiratoires, souventresponsables d'infections chez les jeunes enfants, les personnes âgées et également les patients immunodéprimés. Ces virus possèdent, à la surface de leur enveloppe, deux glycoprotéines qui jouent toutes les deux un rôle dansl’entrée du virus dans la cellule-cible. L'hémagglutinine-neuraminidase (HN) permet au virus de s'attacher aurécepteur cellulaire. Une fois liée au récepteur, HN "active" la seconde glycoprotéine, la protéine de fusion (F). Cette dernière réalise la fusion entre l'enveloppe du virus et la membrane cellulaire. Ce travail de thèse s’estfocalisé principalement sur les glycoprotéines d’enveloppe du virus parainfluenza humain de type 2 (hPIV-2)ainsi que celles d’un virus animal très proche, le virus parainfluenza de type 5 (PIV-5). Une première partie de ce projet a consisté à caractériser des souches circulantes hPIV-2 au niveau de leursglycoprotéines et analyser les différences observées au niveau des domaines fonctionnels des glycoprotéines. Cette étude a notamment souligné l’importance de la mise à jour des outils diagnostiques dans la surveillance deces virus respiratoires. Une seconde partie du projet a consisté à étudier une des étapes du mécanisme d’entrée des virus parainfluenza,La fusion membranaire induite par la glycoprotéine F, dans le modèle viral PIV-5. Ce travail a montré qu’il étaitpossible dans une certaine mesure, par une approche de mutagenèse combinatoire, de dissocier lescaractéristiques de F PIV-5 et ainsi pouvoir mieux les étudier. L’ingénierie d’une glycoprotéine virale telle quePIV-5 pourrait contribuer au développement d’outils en thérapeutique. La description en cryo-microscopie électronique de PIV-5 a été réalisée au cours de ce travail. Ce travail a révéléque la morphologie de PIV-5, dans un état proche de son état naturel, était bien moins hétérogène que ce qui étaitgénéralement décrit dans la littérature. L’analyse d’image a également permit de calculer la densité desglycoprotéines à la surface du virus, donnée nouvelle et intéressante, en regard des connaissances actuelles sur lemécanisme d’entrée des virus parainfluenzaHuman parainfluenza viruses (hPIV, family Paramyxoviridae) are respiratory viruses, often responsible ofinfections at the young children, the elderly and also the immuno-deficient patients. These viruses possess twoglycoproteins at the surface of their envelope, that both play a role in the viral entry into the target cell. Thehemagglutinin-neuraminidase glycoprotein (HN) allows the virus to become attached to the cellular receptor. Then, HN "activates" the second glycoprotein, the protein of fusion (F). This last achieves the fusion betweenthe envelope of the virus and the cellular membrane. The mechanism by which the HN protein "activates" the Fprotein remains unknowns, despite the recent determination of the glycoprotein structures. Several models arecurrently proposed in the literature. This thesis work was mainly focused on the human parainfluenza virus type 2 (hPIV-2) and the animalparainfluenza virus type 5 (PIV-5) envelope glycoproteins. A first step of this project has consisted to characterize several circulating hPIV-2 strains at the level of theirglycoproteins and to analyze the differences observed in their functional domains. This work has underlined theimportance of the diagnostic tools update especially in the surveillance of these respiratory viruses. A second part of the project has consisted to study one of viral entry steps of parainfluenza viruses, themembrane fusion led by the F glycoprotein, in a PIV-5 viral model This work has shown that it was to a certainextent possible, by a combinative mutagenesis approach, to dissociate the features of F PIV-5. The engineeringof a viral glycoprotein like PIV-5 F could contribute to the development of tools in therapeutic. The first description of PIV-5 in cryo-electron microscopy has been achieved during this work. This work hasrevealed that the morphology of PIV-5, in a state close to its fully hydrated natural state, was less heterogeneousthan what was generally described in the literature. It was possible to calculate the surface density of theglycoproteins, a new and interesting data, regarding to the actual knowledge on parainfluenza viruses viral entrymechanisms. Works done during this thesis were also accompanied by the development of diagnostic and therapeuticapplications
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Wheatley, Nicola. "Mapping the haemagglutinin and neuraminidase functions of the human parainfluenza virus type 3 haemagglutinin-neuraminidase protein." Thesis, University of Ottawa (Canada), 1991. http://hdl.handle.net/10393/7739.
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The haemagglutinin-neuraminidase protein (HN) of human parainfluenza virus type 3 is a bifunctional protein. To map the functions to the protein, three truncations of the gene were constructed by digesting the HN gene with the restriction endonucleases Hind III, Bgl II or Xbo I and inserting a stop codon. An internal deletion using Rsa I was also constructed. The four mutants were expressed in the recombinant vaccinia virus system. The expression of the mutant proteins was analysed by both Western Blot and immunoprecipitation. The products of the three truncations migrated at the molecular weights predicted for the truncated proteins. The product of the internal deletion migrated more quickly than predicted and upon sequencing revealed a frame shift mutation and, therefore, a fourth truncation. The migration of the gene product was consistent with the molecular weight predicted for this truncation. The full length HN was functional in both haemagglutination and neuraminidase assays. The four mutants were active only in the neuraminidase assay, none of them haemagglutinated Guinea pig erythrocytes. This allows us to predict that the haemagglutination region is near the carboxy-terminus of the protein while the neuraminidase region is amino-terminal of amino acid 212.
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Hartmann, Tamahine Larronda Schmidt. "Anticorpos neutralizantes contra os vírus da cinomose e parainfluenza caninos em cães e felinos silvestres em cativeiro." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2006. http://hdl.handle.net/10183/8197.
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O vírus da cinomose canina (CDV) e o vírus parainfluenza canino (CPIV) afetam uma ampla variedade de hospedeiros e encontram-se distribuídos mundialmente. O CDV é considerado um dos mais importantes agentes infecciosos dentro das populações caninas. Este vírus é o agente causal da cinomose, uma doença potencialmente letal em membros das famílias Canidae, Mustelidae e Procionidae, sendo recentemente detectado como causa de morbidade e mortalidade em carnívoros aquáticos e grandes felinos. O CPIV, por sua vez, é altamente contagioso entre cães, podendo infectar roedores e gatos em infecções experimentais. Geralmente, o CPIV produz uma traqueobronquite aguda auto-limitante, porém pode atuar sinergicamente com outros agentes infecciosos, como o CDV, causando sinais clínicos mais graves. Como em nosso meio são escassas as informações sobre estes vírus, o presente estudo visou aprofundar os conhecimentos sobre a prevalência de CDV e CPIV em cães e felinos silvestres mantidos em cativeiro. Para tanto, soros destes animais foram testados em busca de anticorpos neutralizantes contra amostras padrão do CDV (Rockborn e Snyder Hill) e do CPIV (V660). Inicialmente, foram testados soros de 173 cães de rua mantidos em canis municipais em Novo Hamburgo e Porto Alegre, RS. A prevalência de anticorpos neutralizantes anti-CDV frente às amostras de vírus da cinomose Rockborn e Snyder Hill, foi de 9,3 % e 4,1 %, respectivamente. Somente dois cães apresentaram títulos de anticorpos considerados protetores contra CDV Rockborn (igual ou maior que 100) e nenhum soro apresentou título de anticorpos neutralizantes considerado protetor para a amostra Snyder Hill (igual ou maior que 100). Contra a amostra de parainfluenza canino V660, a prevalência de anticorpos neutralizantes encontrada foi de 51,4 %. Conclui-se, portanto, que a população de cães de rua amostrada apresenta poucos indícios de contato prévio com CDV, sugerindo grande susceptibilidade à cinomose. Por outro lado, o CPIV parece circular amplamente nesta população. Na segunda parte do presente estudo, como no Brasil não existem relatos sobre CDV e CPIV em felinos silvestres, buscou-se verificar a possibilidade da ocorrência dessas infecções em felinos silvestres brasileiros. Para tanto, foram testados soros de 84 felinos silvestres de seis diferentes espécies nativas do Brasil (Leopardus tigrinus, Puma concolor, Leopardus wiedii, Herpailurus yaguarondi, Panthera onca), todos mantidos em cativeiro em criatórios de distintas regiões do País. Todos os felinos amostrados apresentaram-se soronegativos frente às amostras de CDV e CPIV utilizadas. Estes resultados indicam que CDV e CPIV parecem não circular nas populações de felinos silvestres amostradas.Canine distemper virus (CDV) and canine parainfluenza virus (CPIV) infect a great variety of hosts ranges and are distributed worldwide. CDV is one of the most important infectious agents in dogs. This virus may cause potentially lethal disease among members of the Canidae, Mustelidae and Procionidae families. It has also caused diseases of significant morbidity and mortality in aquatic carnivores and large felids. CPIV, on its turn, is highly contagious among dogs, whilst rodents and cats are susceptible to experimental infections. CPIV is usually associated with an acute selflimiting tracheobronchitis. However, it can act sinergistically with other infectious agents, such as CDV, and cause clinical signs of variable severity. As information on CDV and CPIV infections in our millieu are scarce, this study was carried out aiming to increase knowledge on the prevalence of CDV and CPIV in stray dogs as well as in captive Brazilian wild felids. In order to have an estimate on such prevalences, sera from these animals were tested for neutralizing antibodies to CDV strains Rockborn and Snyder Hill, and to CPIV strain V660. Initially, 173 sera from stray dogs kept in kennels from the municipalities of Novo Hamburgo and Porto Alegre, RS, were examined. The prevalences of neutralizing antibodies to CDV strains Rockborn and Snyder Hill were 9.3 % and 4.1 %, respectively. Only two dogs had antibody levels which could be correlated to protection (that is, titre ≥ 100) to CDV Rockborn whereas no sera presented antibody titres high enough to be considered protective to CDV strain Snyder Hill (that is, titre ≥ 100). Regarding CPIV, the prevalence of anti-V660 neutralizing antibodies was 51.4 %. It can be concluded that the stray dog populations under study shows few serological evidence of previous contact with CDV and seem largely susceptible to CDV infections. On the other hand, CPIV seems to circulate widely in the examined population. In the second part of this study, as there are no reports on CDV and CPIV infections in wild felids in Brazil, it was aimed to determine whether there would be any evidence of such infections among some of such species. For that, 84 sera from wild felids of six different Brazilian native species (Leopardus tigrinus, Puma concolor, Leopardus wiedii, Herpailurus yaguarondi, Panthera onca), all kept in captivity in different regions of the country, were tested for neutralizing antibodies to both CDV and CPIV. All wild felid sera tested were negative for antibodies to the two strains of CDV as well as to CPIV. These results indicate that CDV and CPIV do not seem to circulate among the wild felid populations examined.
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Ocadaque, Crister JosÃ. "Clinical and epidemiological aspects of children pneumonia associated with four types of parainfluenza virus in Fortaleza-CE." Universidade Federal do CearÃ, 2016. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=16327.
Full textAbstract:
FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgicoAs pneumonias sÃo problemas de saÃde publica mundial, especialmente em crianÃas menores que cinco anos de idade. Os vÃrus parainfluenza (VPI-1, 2 e 3) sÃo agentes frequentes de pneumonia, pouco se conhecendo sobre a participaÃÃo do VPI-4 devido a dificuldades do seu isolamento em cultura de cÃlulas, a ausÃncia de antÃgenos especÃficos para este vÃrus nos painÃis de rotina de detecÃÃo dos vÃrus respiratÃrios, alÃm de serem relacionados apenas a casos de infecÃÃes respiratÃrias leves. O objetivo do presente estudo à descrever o perfil epidemiolÃgico e clÃnico das pneumonias causadas pelos quatro tipos de VPI na populaÃÃo de estudo, no perÃodo de janeiro de 2013 a dezembro de 2014. Para tanto, aspirados nasofarÃngeos de 542 crianÃas de atà cinco anos atendidas no Hospital Infantil Albert Sabin (HIAS), que receberam o diagnÃstico de pneumonia, foram submetidos à RT-PCR para a detecÃÃo dos VPI-1, 2 e 3 e 4. Estas amostras tinham sido analisadas anteriormente por imunofluorescÃncia indireta para vÃrus sincicial respiratÃrio (VSR), influenza (A e B), adenovÃrus e VPI (1, 2 e 3). Os VPI foram detectados em 165 casos, seguido de VSR (136), adenovÃrus (34) e influenza (30). As caracterÃsticas clÃnicas e epidemiolÃgicas de pneumonias pelos VPI foram analisadas em 104 amostras que apresentaram infecÃÃo isolada por um dos quatro tipos de VPI. Os VPI mais frequentemente detectados, em ordem decrescente, foram os tipos VPI-3 (64,42%), VPI-4 (19,23%), VPI-1(14,42%) e VPI-2 (1,92%). O VPI-4 foi o mais associado a co-infecÃÃes. O VPI-4 foi o Ãnico VPI cuja circulaÃÃo esteve associada à estaÃÃo chuvosa dos dois anos de estudo (p<0,0001). O VPI-3 e o VPI-1 apresentaram pico de circulaÃÃo associado à estaÃÃo seca. Os VPI sÃo agentes frequentes de pneumonias em crianÃas menores que cinco anos na cidade de Fortaleza.
Pneumonia are public health problems worldwide, especially in children younger than five years old. Human parainfluenza virus (HPIV-1, 2 and 3) are common agents of pneumonia, little was know about the involvement of HPIV-4 due to difficulties of isolation in cell culture, the absence of antigens specific for this virus in panels routine detection of respiratory viruses, and are associated only with cases of mild respiratory infections. The aim of this study is to describe the clinical and epidemiological profile of pneumonia caused by four types of HPIV in the study population, from January 2013 to December 2014. To this end, nasopharyngeal aspirates of 542 children under five treated at Hospital Infantil Albert Sabin (HIAS), who were diagnosed with pneumonia, were subjected to RT-PCR for the detection of HPIV-1, 2, 3 and 4. These samples had been previously analyzed by indirect immunofluorescence for respiratory syncytial virus (RSV), influenza (A and B), adenovirus, and HPIV (1, 2 and 3). The HPIV were detected in 165 cases, followed by RSV (136), adenovirus (34) and influenza (30). Clinical and epidemiological characteristics of pneumonia by HPIV were analyzed in 104 samples with isolated infection with one of four types of HPIV. The HPIV most frequently detected, in descending order, were the HPIV-3 types (64.42%), HPIV-4 (19.23%), HPIV-1 (14.42%) and HPIV-2 (1.92 %). The HPIV-4 was the most associated with co-infection. The HPIV-4 was the only HPIV whose circulation was associated with the rainy season of two years of study (p <0.0001). The HPIV-3 and HPIV-1 had a circulation peak associated with the dry season. The HPIV are frequent agents of pneumonia in children younger than five years in the city of Fortaleza.
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Gonzalez, Carlos M. "Immunopathological studies in the ovine lung during the course of natural and experimental parainfluenza type 3 virus infection." Thesis, University of Edinburgh, 1996. http://hdl.handle.net/1842/30213.
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The pulmonary immunopathology of parainfluenza type 3(PIV-3) infection in sheep was investigated firstly by isolating the virus from field cases of sheep pneumonia, secondly by experimentally reproducing the disease with the isolated virus and finally by studying changes in lymphocytes subsets and alveolar macrophages, induced by PIV-3 in vivo and in vitro. Three ovine virus isolates(270-7, 390-10 and 430-7) were obtained and characterised, as PIV-3, according to virus morphology, under transmission electron microscopy(TEM); cytopathic effect(CPE); haemagglutination, of guinea pig erythrocytes; physicochemical properties; serological crossreactivity with antisera raised against PIV-3, isolated from different species; and reactivity with monoclonal antibodies anti PIV-3 structural proteins, crossreacting with human and bovine strains. The ability of the virus to induce respiratory disease was investigated by experimental inoculation of ovine PIV-3 isolate, 270-7, in colostrum deprived lambs. Clinical, pathological, bacteriological and virological studies were carried out on days 2,3,5 and 7 infection(p.i.). This PIV-3 ovine strain was able to induce clinical disease. Histopathological findings were interstitial pneumonia with hyperplasia of bronchiolar associated lymphoid tissue (BALT), degenerative bronchiolar epithelium with lymphocyte infiltration, areas of atelectasis and increased alveolar septa thickness due to proliferation of pneumocytes II, lymphocytes, macrophages and later to fibrosis. The average cell count per normal BALT was 1,307(range 1,205 to 1,480 cells) whereas in infected lungs on PID 7 the average count increased to 4,788 (range 4,390 to 4,930 cells). The large number of lymphocytes, particularly on days 5 and 7 p.i., combined with the minimal to moderate cytolysis of antigen bearing cells, suggests that PIV-3-induced pulmonary disease has an immunopathological component. PIV-3 particles were detected, by immunohistochemistry, more frequently in bronchiolar epithelium cells than in alveolar septal cells and macrophages by day 3 p.i., but the situation was opposite by day 7 p.i.
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Montesinos, Paredes Milagros de Jesús. "Anticuerpos contra el virus de la Parainfluenza 3 en cerdos de crianza tecnificada y no tecnificada beneficiados en dos mataderos de Lima." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2010. https://hdl.handle.net/20.500.12672/15628.
Full textAbstract:
Publicación a texto completo no autorizada por el autorEl documento digital no refiere asesor
El virus de la parinfluenza 3 (VPI3), es uno de los agentes virales involucrados en el complejo respiratorio que afecta principalmente a los bovinos y otras especies. El objetivo del presente estudio fue determinar la seroprevalencia de este agente viral en porcinos del valle de Lima beneficiados en dos mataderos de la ciudad de Lima. Para este fin se colectaron muestras de suero de porcinos de ambos sexos entre 2 a más de 6 meses de edad provenientes de granjas tecnificadas (n= 192) y de porcinos criados sin o con escasa tecnología (n= 192) para la detección de anticuerpos contra el VPI3 mediante la técnica de neutralización viral. El 5.5±2.3% (21/384) de los porcinos tuvieron anticuerpos contra el VPI3. De este total, el 5.2±1.1% (10/192) y 5.7±1.2% (11/192) de los porcinos de crianza tecnificada y no tecnificada respectivamente, presentaron anticuerpos contra el VPI3. Los títulos de anticuerpos tuvieron un rango de 4 a 64 y fueron detectados en animales mayores de 3 meses de edad. Los resultados indican que el VPI3 no estuvo involucrado en problemas respiratorios en los porcinos muestreados en 2008.
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Rai, Vijeta. "Screening of large collection of compounds for anti-human parainfluenza virus type-2 activity and evaluation of hit compounds." Thesis, Högskolan i Skövde, Institutionen för biovetenskap, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-14385.
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Human parainfluenza virus type-2 (HPIV-2) is a highly contagious respiratory pathogen that can cause severe respiratory disease known as laryngotracheobronchitis or croup-like disease in children. No specific vaccine or an antiviral drug is currently approved for treatment of HPIV-2 infections. In this project, a library of 14400 diverse compounds had been screened for anti-HPIV-2 activities in cultures of African green monkey kidney cells. All compounds that inhibited the virus induced syncytium-forming activity in these cells were considered as hit compounds. Three hit compounds showed moderate anti-HPIV-2 activity characterized by the IC50 values of 20 µM and selectivity indices of approximately 5. This suggests that the antiviral activity of these compounds was due to targeting activities of cellular rather than viral components. Another hit compound, referred to as compound 5, showed anti-HPIV-2 activity that was manifested as a reduction of area of the virus-induced plaques in cells at not cytotoxic concentrations. Interestingly, this compound did not inhibit initial infection nor the virus production in infected cells as revealed by the time-of-addition assay. Moreover, it showed no direct the virus-inactivating (virucidal activity) against HPIV-2 particles. However, relatively short pre-treatment (4 hours) of the cells with compound 5 prior to the virus infection was sufficient for its plaque size-reducing activity suggesting that anti-HPIV-2 activity of compound 5 was due to targeting activities of cellular rather than viral components. Further studies are needed to elucidate the anti-HPIV-2 mechanism of activity of hit compounds identified in the present study.
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FÃ, Mariana Mota Moura. "Perfil clÃnico-epidemiolÃgico das infecÃÃes respiratÃrias agudas causadas por vÃrus parainfluenza em crianÃas atendidas em um hospital de referÃncia da cidade de Fortaleza â CE." Universidade Federal do CearÃ, 2007. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=715.
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CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel SuperiorAs infecÃÃes respiratÃrias agudas (IRAs) sÃo um importante problema de saÃde pÃblica em todo o mundo, e os vÃrus parainfluenza estÃo entre os seus agentes mais freqÃentes. Este estudo teve como objetivos: determinar a freqÃÃncia de IRAs pelo vÃrus parainfluenza entre crianÃas atendidas no Hospital Infantil Albert Sabin, hospital pediÃtrico de referÃncia da cidade de Fortaleza â CE, de janeiro de 2001 a dezembro de 2006; descrever o padrÃo de sazonalidade e as caracterÃsticas clÃnico-epidemiolÃgicas destas infecÃÃes; e comparar as caracterÃsticas clÃnico-epidemiolÃgicas das infecÃÃes por parainfluenza com as das IRAs causadas por outros vÃrus. Foram coletados aspirados de nasofaringe de crianÃas com sintomas de IRAs, e foi utilizada a imunofluorescÃncia indireta para a detecÃÃo dos vÃrus: parainfluenza humano 1, 2 e 3 (VPIH-1, 2 e 3), vÃrus sincicial respiratÃrio (VSR), influenza A e B e adenovÃrus. Nos seis anos de estudo, foram colhidas amostras de 3070 crianÃas, a maioria delas previamente sadias, com a detecÃÃo de vÃrus respiratÃrios em 933 (30,39%), e dos vÃrus parainfluenza em 117 casos (3,81% do total). Dentre os casos de parainfluenza, o VPIH-3 foi o tipo mais freqÃente (83,76% dos casos), com menor detecÃÃo do VPIH-1 (11,96%) e do VPIH-2 (4,27%). A infecÃÃo pelo VPIH-3 apresentou comportamento sazonal, com maior detecÃÃo nos meses de setembro a novembro. Embora o total de casos de IRAs tenha apresentado relaÃÃo direta com os Ãndices pluviomÃtricos, o nÃmero de casos de VPIH-3 apresentou relaÃÃo inversa com a pluviometria, sendo maior nos meses secos. A maioria dos pacientes positivos para parainfluenza foi atendida na emergÃncia ou nos ambulatÃrios. A mÃdia de idades das crianÃas com infecÃÃo pelo VPIH-3 foi de 20 meses, sendo significativamente menor que a das crianÃas infectadas pelo vÃrus influenza A (34 meses), e maior que a das infectadas pelo VSR (15 meses). Os pacientes positivos para o VPIH-3 apresentaram significativamente menos dispnÃia, tosse, estertores, tiragem intercostal e alteraÃÃes radiolÃgicas que os positivos para VSR. As infecÃÃes de vias aÃreas superiores constituÃram a sÃndrome clÃnica mais freqÃente entre os casos de VPIH-3. Os pacientes positivos para VPIH-3 necessitaram menos de terapia com antibiÃticos, corticÃides, oxigÃnio, nebulizaÃÃo e/ou salbutamol que os positivos para VSR. Os resultados demonstraram um comportamento sazonal do VPIH-3, relacionado aos meses secos, o que nÃo tinha sido relatado previamente no Nordeste brasileiro, alÃm de apontarem para uma menor gravidade das infecÃÃes causadas pelo VPIH-3, na comparaÃÃo com o VSR, em crianÃas previamente sadias
Acute respiratory infections (ARI) are an important public health problem throughout the world and parainfluenza viruses are among the major etiologic agents. The objectives of this study were: a) to determine the frequency of parainfluenza infections among children attending Hospital Infantil Albert Sabin, a major pediatric hospital in Fortaleza - CE, from January 2001 to December 2006; b) to describe the seasonal pattern and the clinical and epidemiological characteristics of these infections; and c) to compare clinical and epidemiological characteristics of parainfluenza infections and infections caused by other respiratory viruses. Nasopharyngeal aspirates from children with acute respiratory symptoms were collected and submitted to indirect immunofluorescence assays to detect human parainfluenza virus 1, 2 and 3 (HPIV-1, 2 and 3), respiratory syncytial virus (RSV), influenza A and B and adenovirus. During the six-year study period, samples were collected from 3,070 generally healthy children and respiratory viruses were demonstrated in 933 cases (30.39%), of which 117 were positive for parainfluenza virus (3.81%). HPIV-3 was the most frequently detected type of parainfluenza virus accounting for 83.76% of cases, followed by HPIV-1 (11.96%) and HPIV-2 (4.27%). HPIV-3 infections were seasonal with most cases observed from September to November. Although the total number of ARIs was directly associated with the time of the rainy season, HPIV-3 infections were inversely related with rainfall indices. Most HPIV-3 infections were seen in outpatients. The mean age of patients infected by HPIV-3 was 20 months, which is significantly younger than for influenza A (mean age: 34 months) and significantly older than for RSV (mean age: 15 months). HPIV-3 patients presented significantly lower indices of dyspnea, cough, crackles, chest retractions and radiologic abnormalities than RSV patients. Upper airway infection was the most frequent clinical syndrome among HPIV-3 patients. HPIV-3 patients needed less oxygen, salbutamol, antibiotics, corticosteroids and nebulization than RSV patients. In contrast with earlier observations for Northeastern Brazil, our results demonstrate a seasonal pattern for the occurrence of HPIV-3 infections with most cases observed during the dry season. The results also suggest that infections caused by HPIV-3 are milder than infections caused by RSV in previously healthy children
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Sánchez, Salazar Manuel Rodolfo. "Permisibilidad de cultivos celulares secundarios de alpaca y llama a multiplicación viral de herpesvirus bovino, virus de la diarrea viral bovina, virus parainfluenza 3 bovina y virus respiratorio sincitial bovino." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2006. https://hdl.handle.net/20.500.12672/705.
Full textAbstract:
El objetivo del presente estudio fue determinar la permisibilidad de los cultivos celulares secundarios de alpaca y llama a la infección por distintos agentes virales de conocida seroprevalencia en este tipo de ganado. Se establecieron dos líneas celulares de cornete nasal y piel de alpaca y llama infectándose con Virus de la diarrea viral bovina (VDVB), Virus Herpes Bovino tipo 1 (VHB-1), Virus respiratorio Sincitial Bovino (VRSB) y Virus Parainfluenza bovino tipo 3(VPI-3). Se determinó y caracterizó la presentación de efectos citopatogénicos (ECP) por medio de microscopia óptica de las monocapas teñidas con Hematoxilina-Eosina (HE). Se confirmó la presencia de antígenos virales por medio de la prueba de Inmunofluorescencia Directa (IF). Los cultivos celulares secundarios de piel y cornete nasal de llama y alpaca fueron permisibles a la infección de los distintos virus, presentando los ECP característicos de cada uno de ellos. Esto demuestra que las células de alpaca y llama cultivadas in vitro presentan receptores homólogos a los presentes en células bovinas y determina que este tipo de cultivos es un modelo apropiado para ensayos de infección viral.
Palabras Claves: alpacas, llamas, líneas celulares, permisibilidad, Virus de la diarrea viral bovina (VDVB), Virus Herpes Bovino tipo 1 (VHB-1), Virus respiratorio Sincitial Bovino (VRSB) y Virus Parainfluenza bovino tipo 3(VPI-3), Efectos citopatogénicos (ECP).--- In order to determine the permisibility of alpaca and llama cell cultures to infection by various viral agents of known seroprevalence, nasal turbinate and skin cell lines of alpaca and llama were established and infected with Bovine Viral Diarrhoea Virus (BVDV), Bovine Herpes Virus type 1 (BHV-1), Bovine respiratory Sincitial Virus (BRSV) and Bovine Parainfluenza Virus type 3 (BPIV-3). Presentation of citopathogenic effect (CPE) was determined and characterized by optical microscopy of Hematoxilin-Eosine stained monolayers. The presence of viral antigen was confirmed by Direct Immunofluorescence. Every cell line was permisible to infection with the four viral strains, showing the characteristic CPE. These results prove that alpaca and llama cells cultured in vitro show homologue receptors to those found in bovine cells and determine that these type of cultured cells repesent an appropriate model for viral infection assays. Key Words: alpaca, llama, cell lines, permisibility, Bovine Viral Diarrhoea Virus (BVDV), Bovine Herpes Virus type 1 (BHV-1), Bovine respiratory Sincitial Virus (BRSV) and Bovine Parainfluenza Virus type 3 (BPIV-3), Citopathogenic effect (CPE).
Tesis
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Silva, Bruno Toledo. "Influência dos anticorpos maternos na resposta imune induzida pela vacinação em bezerros Holandeses." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/10/10136/tde-24032016-151529/.
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O objetivo geral desta pesquisa foi avaliar a transferência de imunidade passiva e a sua influência na resposta vacinal para as viroses envolvidas na Doença Respiratória Bovina (DRB). Os dados obtidos nesta pesquisa estão apresentados em dois capítulos. Capítulo 1 - Objetivou-se avaliar a dinâmica de anticorpos (Acs) específicos para as viroses respiratórias e subpopulações de linfócitos em bezerros do nascimento aos 240 dias (d) de vida. Para tanto, acompanhou-se a transferência de imunidade passiva de Acs específicos para as viroses respiratórias em 19 bezerros, destes cinco foram selecionados para acompanhamento da dinâmica de Acs neutralizantes e subpopulações de linfócitos dos 14 aos 240d. O colostro fornecido era proveniente de vacas doadoras vacinadas. A análise da qualidade individual do colostro revelou índice Brix ≥21%, observando-se forte correlação desses valores com proteína total (r= 0,942 e P= 0,001). Após 48 horas da ingestão do colostro, pôde-se observar soroconversão dos 19 animais (100%) para os agentes virais envolvidos na DBR. As medianas (Log2) encontradas foram de 12,3, 9,0, 5,0 e 8,5 para BVDV, BoHV-1, BRSV e BPIV-3. Dos 14 aos 240d os 5 bezerros avaliados, demonstraram declínio gradual dos títulos de Acs (Log2) para BVDV (12,8-3,3), BoHV-1 (10,0-3,3) e BPIV-3 (10,0-2,0), embora o BVDV não tenha apresentado soronegatividade até os 240d. Manifestações clínicas de broncopneumonia foram observadas em 4/5 (80%) bezerros dos 80 aos 135 d. BRSV (11,3-2,0) apresentou perfil diferenciado na dinâmica de anticorpos em relação às demais viroses. Não foram detectadas diferenças estatísticas entre os momentos para as viroses apesar das variações detectadas (P>0,05). A meia-vida e tempo para soronegatividade foram de 36,2±6,1 e 367,01±68,7d para BVDV, 50,7±18,0 e 239,67±66,88d para BoHV-1 e, 46,8±21,1 e 303,36±60,15d para BPIV- 3. BRSV não respeitou modelo de regressão para cálculos de meia-vida e soronegatividade. Valores absolutos e relativos das populações de linfócitos não revelaram diferenças estatísticas entre os momentos (P>0,05). Contudo, a dinâmica das subpopulações de linfócitos revelou aumento de células B CD21+ até 150d; aumento nos valores relativos das subpopulações CD4+, CD8+ e CD3+CD4-CD8- principalmente aos 74-90d. Assim, pôde-se determinar uma janela de susceptibilidade a partir dos 74d especialmente ao BRSV e BoHV-1, momento que precede o aumento dos títulos de Acs após exposição natural. Capítulo 2 Objetivou-se avaliar a influência dos Acs maternos na resposta imune para DRB induzida pela vacinação. Foram selecionados 23 bezerros recém-nascidos, distribuídos aleatoriamente entre 4 grupos experimentais: G1 vacinado aos 14d e booster aos 44d; G2 aos 90d e aos 120d; G3 aos 180d e aos 210d. Além disso manteve-se um grupo controle não vacinado CG1, CG2 e CG3. Os bezerros foram vacinados com a mesma vacina comercial empregada para vacinação das doadoras de colostro. Observou-se: (1) a vacina com BVDV inativado não promoveu aumento dos títulos de Acs para nenhum dos grupos avaliados; (2) G1 não demonstrou soroconversão para nenhuma das viroses, enquanto controle CG1 exibiu decréscimo dos títulos para BVDV, BoHV-1 e BPIV-3; (3) o BRSV apresentou baixa soroconversão no G2, enquanto o controle demonstrou altos títulos dos 44-120d; (4) não foi possível distinguir entre quais tempos ocorreram diferenças entre títulos (P>0,0167); (5) linfócitos B (CD21+) aumentaram do T0-T2 para G1, diminuíram para G2, e aumentaram no G3; (6) linfócitos T CD3+ diminuíram ao longo do tempo para todos os grupos, exceto CG3; (7) apesar de oscilações, linfócitos T CD4+, CD8+ e WC1+ se mantiveram praticamente constantes até 240d, exibindo maiores proporções nos grupos vacinados; (8) a expressão do marcador CD25+foi mantida pelo grupo G1 até o T2, mas apresentou aumento no G2 e G3; (9) manifestações de broncopneumonias foram identificadas nos bezerros do grupo controle (4/5 - 80%) e podem ter exercido influência nas diferenças encontradas para as células entre os grupos. Em geral, a vacinação dos bezerros aos 90 (G2) e 180d (G3) manteve ou estimulou a produção de Acs para o BoHV-1, BRSV e BPIV-3, e a ativação das células T expressas pelo marcador CD25+ pode ter sido responsável pela proteção dos bezerros frente à DRB. Assim, com base nos resultados, concluiu-se que a intensidade da imunidade dos bezerros induzida pela vacinação aumentou de acordo com o desenvolvimento etário e diminuição dos títulos de Acs maternos. A conclusão geral desta dissertação aponta para a necessidade precoce de imunização dos bezerros, especialmente pela susceptibilidade observada para BRSV e BPIV-3 aos 74-90d de vida. Entretanto, esta pesquisa não encontrou resposta humoral induzida pela vacinação no grupo de bezerros vacinados aos 14 e 44 dias, apesar dos indícios de resposta imune celular. Assim, estudos futuros devem ser elaborados considerando estratégias para amplificar a resposta imune precoce dos bezerros para os agentes virais envolvidos na DRBThe main purpose of this research was to evaluate the transfer of passive immunity and its influence on vaccine response to the viruses involved in bovine respiratory disease (BRD). The data obtained in this study are presented in two chapters. Chapter 1 - The objective was to evaluate the dynamics of specific antibodies to the respiratory viruses and lymphocyte subpopulations in the calves from birth to 240 days (d) of life. Thus, the transfer of passive immunity of specific antibodies to the respiratory viruses were assessed in 19 calves; from these, five were selected for monitoring the dynamic of neutralizing Abs and lymphocytes subpopulations from 14 to 240d. The colostrum was provided from donor vaccinated cows. The analysis of individual quality of the colostrum revealed Brix ≥21%, observing strong correlation of these values with total protein (r = 0.942 and P = 0.001). After 48 hours of colostrum intake, was observed seroconversion of the 19 animals (100%) for the viral agents involved in the DBR. The median (Log2) ratio found was 12.3, 9.0, 5.0 and 8.5 for BVDV, BoHV-1, BRSV and BPIV-3. The 5 calves followed from 14 to 240d showed gradual decline in antibody titers (Log2) to BVDV (12.8 to 3.3), BoHV-1 (10.0 to 3.3) and BPIV-3 (10, 0-2.0), although could not be detected seronegative calves for BVDV up to eight months of age. Clinical manifestations of bronchopneumonia were observed in 4/5 (80%) calves from 80 to 135 days of life. BRSV (11.3 to 2.0) showed a distinct profile in the dynamics of antibodies compared to other viruses. There were no statistical differences between times for viruses despite variations detected (P> 0.05). The half-life and time to become seronegative were 36.2±6.1 and 367.01±68.7d for BVDV, 50.7±18.0 and 239.67± 66.88d for BoHV-1 and, 46.8±21.1 and 303.36±60.15d for BPIV-3. BRSV did not respect regression model to perform half-life and seronegative calculations. Absolute and relative values of lymphocyte populations revealed no statistical differences between times (P> 0.05). However, the dynamics of lymphocyte subpopulations showed increase in B cells CD21+ up to 150d; increase in the relative values of CD4+, CD8+ and CD3+CD4-CD8- subpopulations, mainly to 74-90d. Thus, it was possible to determine a window of susceptibility since 74d especially for BRSV and BoHV-1, moment that precede the increase in antibody titers after natural exposure. Chapter 2 - The objective was to evaluate the influence of maternal antibodies in the immune response to respiratory viruses induced by vaccination. Was selected 23 newborn calves that were randomly distributed in four groups: G1 - vaccinated at 14d and booster at 44d; G2 - vaccinated at 90d and booster at 120d; G3 - vaccinated at 180d and booster at 210d. Furthermore were kept a non-vaccinated control group - CG1, CG2 and CG3. Calves were vaccinated with the same commercial vaccine in the colostrum donors. From these, could be observed: (1) the vaccine with inactivated BVDV did not promote increase of antibodies titers in any of the assessed groups; (2) G1 did not demonstrated seroconversion for any of the viruses while CG1 control exhibited decrease in the titers for BVDV, BoHV-1 and BPIV-3; (3) BRSV had low seroconversion in G2, while the control showed high titers from 44 to 120d; (4) where the differences between times occurred could not be distinguished (P <0.0167); (5) B cells (CD21+) increased from T0 to T2 for G1, decreased for G2, and increased for G3; (6) T lymphocytes CD3+ decreased over time for all groups except for CG3; (7) despite variations, T lymphocytes CD4+, CD8+ and WC1+ remained almost constant until 240d, displaying greater proportions in the vaccinated groups; (8) the expression of the CD25+ marker was maintained in the vaccinated group G1 up to T2, whereas vaccination promoted an increase of this expression in G2 and G3; (9) clinical manifestations of bronchopneumonia were identified in the control group (4/5 calves - 80%) and may have influenceon the differences found for the cells between the groups. In general, vaccination of calves at 90 (G2) and 180d (G3) maintained or stimulated the production of Abs to BoHV-1, BRSV and BPIV-3, and the activation of T cells expressed by the CD25+ marker may have been responsible for the protection of calves from BRD. Thus, based on the results, it was concluded that the intensity of the immunity induced by vaccination of calves increased according to the age of development and decay of maternal antibody titers. The general conclusion of this research, points to the need for early immunization of calves, especially by the susceptibility observed for BRSV and BPIV-3 from 74-90d of life. However, this research didnot found humoral response induced by vaccination in the group of calves vaccinated at 14 and 44 days, despite the evidence of cellular immune response. Thus, future studies should be designed considering strategies to amplify the early immune response of calves to the viral agents involved in the BRD
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Prinoski, Kevin Andrew. "Envelope associated proteins of human parainfluenza virus 3 I. M (matrix) gene sequence, and II. F (fusion) gene sequence comparison among ten isolates." Thesis, University of Ottawa (Canada), 1989. http://hdl.handle.net/10393/5655.
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Monteiro, Francielle Liz. "Detecção molecular de vírus respiratórios em cães." Universidade Federal de Santa Maria, 2015. http://repositorio.ufsm.br/handle/1/10238.
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Conselho Nacional de Desenvolvimento Científico e TecnológicoThe respiratory viruses of dogs are associated with a disease called canine infectious respiratory disease (CIRD). The main etiological agents of CIRD are canine distemper virus (CDV), canine parainfluenza virus (cPIV), canine adenovirus type 2 and canid herpesvirus type 1 (CaHV-1), which may cause single or mixed infections. CIRD occurs most frequently in places with high animal density and constant movement. CDV, cPIV, CAdV-2 and CaHV-1 infections have been described worldwide, however, few reports of molecular identification of these viruses are available in Brazil. Thus, the objective of this study was to investigate the occurrence of respiratory viruses in dogs in Santa Maria, RS, and in dog shelters in RS, trying to correlate their occurrence with the environmental conditions. Nasal secretions were collected from dogs with respiratory signs submitted to veterinary clinics in Santa Maria; and from dogs of three shelters of RS (Cachoeira do Sul [shelters #1 and #2] and Passo Fundo [shelter #3]). Viral detection/identification was performed by polymerase chain reaction (PCR) for CDV, cPIV, CAdV-2 and CaHV-1. Positive samples were sequenced and, for some viruses, phylogenetic analysis was performed, comparing with sequences deposited in GenBank. Samples of shelters #1 and #3 were obtained during the cold season. Shelter #1 presented poor sanitary and nutrition conditions, high animal density and constant direct contact among dogs. In this shelter 78% (58/74) of the respiratory samples were positive for at least one virus. The single infections were caused by cPIV in 30% (22/74) of the samples and CAdV-2 in 5% (4/74). Coinfections represented 23% (cPIV and CAdV-2); 13% cPIV, CDV and CAdV-2; 4% cPIV-2 and CDV; and 3% CDV and CAdV-2. Shelters #2 and #3 presented satisfactory sanitary and nutrition conditions, with large outdoors exercise areas (#2) and animal separation by groups (#3). In shelter #2, 8% (5/35) of the samples were positive to cPIV and 6% to CaHV-1; in shelter #3, 8% (7/77) of the samples were positive to CAdV-2 and 1% to CDV. Of samples obtained in Santa Maria, 40% (10/25) were positive for virus, being 28% (7/25) for cPIV, and 4% (1/25) to each of the other viruses. Thus, the results obtained demonstrate that infections and coinfections by respiratory viruses are common in shelter dogs in RS, and their occurrence is related to population density, health and nutritional conditions and season. These viruses are also circulating in domestic dogs in Santa Maria, associated with respiratory disease. This study reinforces the importance of preventive measures such as vaccination and good environmental conditions to prevent/reduce infections caused by respiratory viruses in dogs.
Os vírus respiratórios de cães estão associados com uma enfermidade denominada doença respiratória infecciosa canina (canine infectious respiratory disease - CIRD). Os principais agentes da CIRD são o vírus da cinomose (canine distemper virus - CDV), vírus da parainfluenza canina tipo 2 (canine parainfluenza virus - cPIV), adenovírus canino tipo 2 (canine adenovirus type 2 - CAdV-2) e herpesvírus canino tipo 1 (canid herpesvirus 1 - CaHV-1), que podem causar infecções simples ou mistas. A CIRD ocorre com maior frequência em locais com alta densidade populacional e constante fluxo de animais. Infecções pelo CDV, cPIV, CAdV-2 e CaHV-1 tem sido descritas em vários países, contudo, são escassos os relatos da identificação molecular desses agentes no Brasil. Além disso, há falta de estudos relacionados aos fatores que favorecem a ocorrência e disseminação desses agentes em cães de abrigos. Desta forma, o objetivo do presente estudo foi investigar a ocorrência de vírus respiratórios em cães do município de Santa Maria, Rio Grande do Sul, e em cães de abrigos, buscando-se associar a ocorrência das infecções com as condições ambientais. Para isso, foram coletadas secreções nasais de cães com sinais respiratórios em clínicas veterinárias de Santa Maria; e de cães de três abrigos do estado do RS (dois em Cachoeira do Sul [#1 e #2] e um em Passo Fundo [#3]). A identificação viral foi realizada por reação em cadeia de polimerase (PCR) para o CDV, cPIV, CAdV-2 e CaHV-1. As amostras positivas foram sequenciadas e, para alguns vírus, foi realizada a análise filogenética, comparando-se com sequências depositadas no GenBank. As amostras dos abrigos #1 e #3 foram obtidas durante épocas de baixas temperaturas. O abrigo #1 apresentava condições sanitárias e nutricionais precárias, além de alta densidade populacional e constante contato entre os cães. Neste abrigo, 78% (58/74) das amostras foram positivas para, pelo menos, um dos vírus investigados. As infecções simples foram causadas pelo cPIV em 30% (22/74) das amostras e CAdV-2 em 5% (4/74). As coinfecções totalizaram 23% (17/74) para o cPIV e CAdV-2; 13% (10/74) para o cPIV, CDV e CAdV-2; 4% (3/74) para o cPIV e CDV; e 3% (2/74) para o CDV e CAdV-2. Os abrigos #2 e #3 eram higienizados corretamente e os cães recebiam alimentação adequada, sendo que no abrigo #2 os animais possuíam amplo espaço para se exercitarem, e no abrigo #3 os animais eram separados em grupos e alojados em gaiolas. No abrigo #2 foram detectadas 8% de amostras positivas para o cPIV e 6% para o CaHV-1; e no abrigo #3, 8% de amostras positivas para o CAdV-2 e 1% para o CDV. Das amostras obtidas em clínicas de Santa Maria, 40% (10/25) foram positivas para um dos vírus pesquisados, sendo 28% (7/25) para o cPIV, e 4% (1/25) para cada um dos outros vírus. Assim, os resultados obtidos demonstram que infecções e coinfecções por vírus respiratórios são comuns em cães de abrigos no estado do RS, estando relacionadas com a densidade populacional, condições sanitárias e nutricionais e estação do ano. Estes vírus também circulam em cães domésticos em Santa Maria, estando associados com doença respiratória. Este estudo reforça a importância de medidas de prevenção, tendo em vista que a vacinação e boas condições ambientais podem reduzir e/ou prevenir as infecções causadas por vírus respiratórios em cães.
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Victorio, Cisneros Willy Mayk. "Seroprevalencia de los virus neumopatógenos de la Parainfluenza bovina 3 (VPI3), Herpesvirus bovino 1 (HVB1) y Virus respiratorio sincitial bovino (VRSB) en alpacas adultas de la provincia de Canchis - Cusco." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2004. https://hdl.handle.net/20.500.12672/2271.
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Con el objetivo de tener una información general sobre la presencia de agentes virales neumotrópicos, se muestreó 345 alpacas adultas procedentes de 31 hatos, pertenecientes a medianos y pequeños criadores de la provincia de Canchis - Cusco. Las muestras de sueros sanguíneos fueron procesadas para la detección de anticuerpos neutralizantes para los virus: Parainfluenza tipo 3 (PI3), Respiratorio Sincitial Bovino (RSB) y el Herpes virus bovino tipo 1 (VHB1). Todas las muestras fueron trabajadas mediante la prueba de neutralización viral utilizando cultivos celulares secundarios de origen bovino (cornetes nasales) y cepas virales citopáticas mantenidas en el laboratorio. Las pruebas de laboratorio evidencian que los 31 hatos muestreados tienen animales seropositivos a los virus RSB y PI3; y solamente 1/30 se encontró animales reactores al VHB1. El estudio, además determino una seroprevalencia de 80.16% ± 6.96% (101/126) y 67.54% ± 4.94% (233/345) del VRSB y el PI3 respectivamente. Estos resultados sugieren que los animales muestreados han estado expuestos a varios agentes potencialmente neumopatogénicos, y evidencian una activa replicación viral al momento del muestreo que coincidió con una severa ola de baja temperatura en la zona y mortalidad de animales por neumonías en el campo, quedando por demostrarse la presencia directa de estos agentes en muestras patológicas y verificar el posible rol causal en procesos neumónicos.The objective of the present study was determine the presence of respiratory virus. Three hundred forty five serum samples from thirty one farms were tested by virus neutralization test to detect neutralizing antibodies against PI-3, BHV1 and BRSV. The farms were small farms from the province of Canchis - Cusco. All the samples were worked by means of the test of viral neutralization using secondary cellular cultures of bovine origin and cytopathic virus from the laboratory. Thirty one farms had seroreactive animals to BRSV and PI-3; and one farm had seroreactive animals to BHV1. The 80,16% ± 6,96% (101/126) and 67,54% ± 4,94% (233/345) of the samples had antibodies against BRSV and PI-3 respectively. These results suggest that sample animals have been exposed potentially to respiratory virus, and demonstrate an active viral replication at the time of the sampling that agreed with a severe low temperature in the zone and mortality of animals by pneumonia in the field.
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Cabellos, Rojas Karina Olinda. "Seroprevalencia de los virus : parainfluenza 3, respiratorio sincitial, diarrea viral bovina, en un rebaño mixto de una comunidad campensina de la provincia de Calca, Cusco." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2006. https://hdl.handle.net/20.500.12672/684.
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El presente estudio tuvo como objetivo determinar la seroprevalencia de los agentes neumotrópicos como los virus de la Diarrea Viral Bovina (VDVB), Respiratorio Sincitial (VRS) y Parainfluenza 3 (VPI3) en rumiantes de la comunidad de Chahuaytiri, provincia de Calca, Cusco, a través de la detección de anticuerpos en el suero sanguíneo de alpacas (n igual 21), bovinos (n igual 66) y ovinos (n igual 152) mediante la prueba de neutralización viral (NV). El 15.8±16.4% (3/21), 4.8 ± 9.1% (1/21) y el 23.8±18.2% (5/21) de las alpacas presentaron anticuerpos neutralizantes contra los virus: DVB, RS y PI3. El 90.9±6.9% (60/66), 87.88±7.9% (58/66) y el 81.8 ± 9.3% (54/66) de los bovinos y el 28.29±7.1% (43/152), 49.34 ±7.9% (75/152) y 50.0 ±7.9% (76/152) de los ovinos presentaron anticuerpos contra la DVB, RS y PI3, respectivamente. Estos resultados confirman la presencia de infecciones virales en rumiantes bajo sistema de crianza mixto de la comunidad campesina del Cusco.--- The seroprevalence of Bovine Viral Diarrhoea Virus (BVDV), Parainfluenza Virus Type 3 (PI3V) and Respiratory Syncytial Virus (RSV) in serum samples of alpacas (n igual 21), bovine (n igual 66) and sheep (n igual 152) of a rural community of Cusco, Peru. It was carried out by virus-neutralization test. The 15.8±16.4% (3/21), 4.8 ±9.1% (1/21) and 23.8±18.2% (5/21) of alpacas had neutralizing antibodies against BVD, RS and PI3 virus. The 90.9±6.9% (60/66), 87.88±7.9% (58/66) and 81.8 ±9.3% (54/66) of bovine and the 28.29±7.1% (43/152), 49.34 ±7.9% (75/152) and 50.0 ±7.9% (76/152) of sheep had antibodies to BVDV, RSV and PI3V respectively. These results confirm the presence of viral infections in ruminants of mixed breeding system of a rural community.
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Grissett, Gretchen Phoebe. "Systematic review of cattle responses to viral and bacterial bovine respiratory disease pathogens and effect of high ambient temperaure on viral replication and serology to an intranasal modified-live (bovine rhinotracheitis-parainfluenza-3) viral vaccine in beef cattle." Thesis, Kansas State University, 2014. http://hdl.handle.net/2097/18169.
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Master of Veterinary Biomedical SciencesDepartment of Clinical Sciences
Bradley White
Objective- To compare serologic response and viral replication following intranasal administration of a modified-live bovine rhinotracheitis (IBR) parainfluenza-3 (PI-3) vaccine in high (32°C) and moderate (21°C) ambient temperatures. Animals- 28 heifers (mean body weight, 206.8 kg) Procedures- Heifers randomly allocated to treatment groups: High Ambient Temperature (HAT, n=10): received vaccine, housed outdoors, Moderate Ambient Temperature (MAT, n=10): received vaccine, housed indoors, High Ambient Control (HAC, n=4): no vaccine, housed outdoors, Moderate Ambient Control (MAC, n=4): no vaccine, housed indoors. Rectal and nasal mucosal temperatures were recorded every 2 hours from 8am to 8pm on trial days 0 and 1. Nasal swabs were collected on trial days 0 through 7 for virus isolation. Serum samples were collected for serology on trial days 0, 7, 14, and 28. Results- Rectal temperatures did not differ among treatment groups over the study period, but nasal temperatures were higher in the HAT calves compared to MAT group at study hours: 6, 24, 30, 32, and 38. Two weeks post-vaccination, IBR titers were significantly greater in vaccinates (HAT,MAT) relative to non-vaccinates (HAC, LAC), but no differences were identified among HAT and MAT. Viable IBR virus was recovered via virus isolation from all vaccinated calves (HAT,MAT) on trial days 1 through 6. Conclusions and Clinical Relevance- The ability to isolate IBR and stimulate the calf immune response following administration of a modified-live IBR-PI3 intranasal vaccine did not differ in calves housed in temperature-controlled and high ambient temperature environments.
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Paris, Fernanda de. "Epidemiologia dos vírus respiratórios e avaliação das características genéticas do vírus sincicial respiratório entre crianças atendidas no Hospital de Clínicas de Porto Alegre." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2012. http://hdl.handle.net/10183/70397.
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Introdução: As infecções respiratórias causam elevadas morbidade e mortalidade, sendo os vírus os principais agentes destas doenças. O monitoramento e vigilância de vírus respiratórios, desde os mais conhecidos até os emergentes, são importantes para a gestão em saúde, orientando tempo de profilaxia e minimizando o impacto de epidemias nas comunidades. Objetivos: Estudar a epidemiologia molecular do vírus sincicial respiratório (VSR) e descrever a epidemiologia dos seguintes vírus: influenza (IF), influenza A (H1N1), adenovírus (AdV) e parainfluenza (PIV) no Hospital de Clínicas de Porto Alegre. Para isso foram conduzidos três estudos: (1) caracterização das infecções respiratórias causadas por VSR, IF, AdV e PIV em crianças; (2) validação de uma técnica de reação em cadeia da polimerase em tempo real (RT-PCR) para detecção de VSR A/B e IF A/B e (3) caracterização de genótipos do VRS em crianças com infecções comunitárias e hospitalares. Métodos: No primeiro estudo foram levantadas as seguintes variáveis: casos de infecções respiratórias por VSR, AdV, PIV ou IF e H1N1; internações em enfermarias hospitalares e internações em unidades de terapia intensiva (UTI); infecções hospitalares e taxas de letalidade. As variáveis foram coletadas durante o atendimento de crianças (idade 0-12 anos) no HCPA entre 2007 e 2010. No segundo estudo, os alvos do ensaio de RT-PCR foram: o gene da proteína da matriz de IFA, o gene da hemaglutinina do IFB e o gene da nucleoproteína de RSVA/B. A especificidade do ensaio e seu limite de detecção foram determinados. Uma comparação entre RT-PCR e imunofluorescência indireta foi realizada. No terceiro estudo, 63 isolados de VSR (21 de origem nosocomial e 42 adquiridos na comunidade) foram sequenciados para estabelecer uma análise filogenética deste vírus. Resultados: Cada um dos vírus estudados apresentou um comportamento diferente. O VSR demonstrou ser o principal agente envolvido em infecções nosocomiais. Já os pacientes com AdV, bem como o VSR, apresentaram altas taxas de admissão em UTI em 2007 e 2010. O AdV e o H1N1 tiveram as maiores taxas de letalidade. O ensaio de RT-PCR mostrou-se específico e foi mais sensível do que a imunofluorescência, sendo capaz de detectar co-infecções. Os seguintes limites de detecção foram obtidos: 1 cópia/mL para a IFA, 10 cópias/mL para IFB, 5 cópias/mL para RSVA e 250 cópias/mL para RSVB. A investigação dos genótipos de VSR revelou que todos os isolados VSRA circulantes eram do mesmo grupo filogenético, o genótipo NA1 de origem japonesa. Por outro lado, os isolados VSRB foram distribuídos em grupos diferentes: BA4, POA1, POA2, POA3 e POA4. Este estudo foi o primeiro que descreveu a circulação do genótipo NA1 no Brasil, bem como quatro novos genótipos (POA1, POA2, POA3 e POA4). Conclusão: Os resultados obtidos no primeiro estudo demonstram o impacto das epidemias sazonais de vírus respiratórios. O segundo estudo corroborou relatos da literatura que técnicas moleculares, como RT-PCR, são adequadas para a detecção de vírus respiratórios. O terceiro estudo relatou genótipos emergentes de VSR. Estudos de vigilância como os descritos acima deveriam ser conduzidos periodicamente para acompanhar o padrão de comportamento dos vírus na população alvo.Background: Respiratory tract infections of viral origin are a major cause of morbidity and mortality worldwide. Surveillance of well-known viruses and emerging threats is important for management, prophylaxis and to minimize impact on the community. Objective: To study the molecular epidemiology of respiratory syncytial virus (RSV) and describe the epidemiology of viruses: influenza (IF), influenza A (H1N1), adenovirus (AdV) and parainfluenza (PIV) at Hospital de Clinicas de Porto Alegre. Three studies were performed: (1) characterization of respiratory infections caused by RSV, IF, AdV and PIV in children, (2) validation of a technique of polymerase chain reaction in real-time (RT-PCR) to detect RSVA/B and IFA/B and (3) detection of genotypes of RSV in children with communityand hospital-acquired infection. Methods: In the first study, variables such as number of cases of viral (RSV, AdV, PIV or IF and H1N1) infection, hospitalizations in general wards and intensive care units (ICUs), nosocomial infections and lethality rates were collected. These variables obtained from each children (age 0-12 years) between 2007 and 2010. In the second study the assay RT-PCR was used to target the matrix gene of IFA, the hemagglutinin gene of IFB and the nucleoprotein gene of RSVA/B. The specificity of the assay and its limit of detection were determined. A comparison of RT-PCR and indirect immunofluorescence was performed. In the third study, 63 RSV isolates (21 nosocomial and 42 community-acquired) were submitted to sequencing to establish a phylogenetic analysis of this virus. Results: The different viruses presented a diversity of behaviors according to hospitalization, nosocomial outbreaks or lethality in children. RSV accounted for most nosocomial infections. Rates of ICU admission for AdV and RSV infection were highest in 2007 and 2010. During 2008–2009, H1N1 and AdV had the highest ICU admission rates. AdV and H1N1 had the highest lethality rates. The RT-PCR assay was more sensitive than immunofluorescence and it was able to detect viral co-infections. Futhermore, the limits of detection were 1 copy/μL for IFA, 10 copies/μL for IFB, 5 copies/μL for RSVA, and 250 copies/μL for RSVB. The genotyping study showed that hospital- and community-acquired RSVA isolates were from the same phylogenetic group (the same group of the NA1 Japanese isolates). Conversely, RSVB isolates were distributed across different groups: BA4, POA1, POA2, POA3 and POA4. This was the first study to describe circulation of the NA1 genotype in Brazil, as well as four RSVB genotypes: POA1, POA2, POA3 and POA4. Conclusion: The results obtained in the first study demonstrate the impact of seasonal epidemics of respiratory viruses. The second study confirmed that molecular techniques such as RT-PCR, are suitable for the detection of respiratory viruses. The third study reported emerging genotypes of RSV. Surveillance studies such as this should be performed periodically to monitor the behavior pattern of the virus in the target population.
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Caignard, Grégory. "Cartographie des intéractions entre protéines virales codées par le gène P des Paramyxoviridae et protéines cellulaires." Paris 7, 2010. http://www.theses.fr/2010PA077020.
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La famille des Paramyxoviridae comprend plusieurs virus pathogènes pour l'homme, certains connus depuis longtemps comme le virus de la rougeole (MV), le virus des oreillons (MuV) ou le virus parainfluenza de type 3 (hPIV3), et d'autres identifiés plus récemment comme le virus Nipah (NiV). Dans le cadre d'un programme de cartographie des interactions virus-hôte, mon projet de thèse vise à identifier les cibles cellulaires des protéines codées par le gène P de ces quatre virus ainsi que du virus Tioman (TioV) dont le seul hôte connu à ce jour est la chauve-souris frugivore. J'ai ainsi réalisé 44 cribles double-hybride et identifié une centaine d'interactions virus-hôte nouvelles. L'analyse de ces interactions montre clairement un enrichissement pour des facteurs de transduction du signal, notamment des protéines impliquées dans la réponse immunitaire. L'identification de STAT1, STAT2 et Jakl, comme partenaires cellulaires de la protéine V du virus de la rougeole nous a permis d'expliquer l'inhibition de la réponse interféron de type I par cette protéine virale. Par ailleurs, nous avons démontré que la protéine V du virus Tioman n'est pas capable de bloquer efficacement le signal IFN-cc/|3 dans des cellules humaines, probablement à cause d'un défaut d'interaction avec STAT2. Une autre étude nous a permis de mettre en évidence l'activité duale portée par la protéine C du virus hPIV3 sur le signal IFN-ct/|3 et la voie MAPK/ERK. La potentialisation de la voie MAPK/ERK en réponse aux facteurs de croissance comme l'EGF par C-hPIV3 pourrait expliquer Phyper-inflammation de Pépithélium respiratoire observée dans le cas d'une infection par ce virusViruses belonging to Paramyxoviridae family are important human pathogens. Some, such as measles virus (MV), mumps virus (MuV) and human parainfluenza virus type 3 (hPIV3), have been known for a long time. Others, such as Nipah virus (NiV), have been recently identified. As part of a large-scale mapping project of virus-host protein interactions, the goal of my thesis is to identify interactions between viral proteins encoded by the P gene of these four paramyxoviruses and cellular proteins. Tioman virus (TioV), whose natural host is the flying fox, was also included in this project. Therefore, 44 yeast two-hybrid screens were performed and hundreds of new virus-host interactions were identified. The global analysis of these interactions has uncovered many signal transduction factors, in particular proteins involved in immune response. The identification of STAT1, STAT2 and Jakl as interactors of MV-V protein allowed us to explain type IIFN signaling inhibition by this viral protein. Futhermore, we demonstrated that TioV-V protein was unable to block efficiently type I IFN signaling in human cells probably because of its inability to interact with STAT2. Finally, we have shown that hPIV3 C protein both increases MAPK/ERK signaling in response to EGF and inhibits type I IFN signaling. These data suggest that an excessive activation of MAPK/ERK pathway by hPIV3-C contributes to the severe airway inflammation associated with hPIV3 infections
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Hanmod, Santosh S. Hewett-Emmett David Peters Ronald J. Chemaly Roy F. "The burden of parainfluenza virus infection in patients with hematological malignancy and hematopoietic stem cell transplant (HSCT) recipients in the absence of active immunization and approved therapy : the role of infection control." 2009. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:1470186.
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Li, Xiantang. "Effects of 4-ipomeanol on bovine parainfluenza type 3 virus induced pneumonia in calves." 1990. http://catalog.hathitrust.org/api/volumes/oclc/23066867.html.
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Thesis (Ph. D.)--University of Wisconsin--Madison, 1990.Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.
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Mao, Shin-Ting, and 毛詩婷. "Production and analysis of monoclonal antibodies against the fiber protein of canine adenovirus and the HN protein of canine parainfluenza virus." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/07685229383000947843.
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碩士國立中興大學
分子生物學研究所
103
Canine adenovirus (CAV) and canine parainfluenza virus (CPiV) were important pathogens to induce kennel cough. The fiber protein on the CAV coat protein is related to virus entry. The hemagglutinin-neuraminidase protein of canine parainfluenza virus (CPiV) could promote virus entry and productive infection. In this study, we deleted the N-terminal 41 and 94 amino acids of CAV and CPiV, respectively and named as CAV-fiber(d41) and CPiV-HN(d94). According to the antigenicity and hydrophilicity of CPiV-HN(d94), three fragments of aa 65-287, aa 288-480 and aa 142-482 we constructed, and named as CPiV-HN(A), CPiV-HN(B) and CPiV-HN(C), respectively. The expected sizes of the CAV-fiber(d41), CPiV-HN(A), CPiV-HN(B) and CPiV-HN(C) were 75 kDa, 47 kDa, 41 kDa and 58 kDa, respectively, and expressed by the E.coli expression system. BALB/c mice were immunized by respective CAV-fiber(d41) and CPiV-HN(d94) proteins to produce monoclonal antibodies. Antibodies secreted form hybridoma cells were screened by ELISA, and further confirmed by dot blot and Western blot assays. Three monoclonal antibodies were prepared in this study, including two anti-CAV-fiber(d41) and one anti-CPiV-HN(d94). Immuno dot and Western blot analyses suggested that both CPiV 72 and CAV 33 monoclonal antibodies recognized a linear epitope and CAV 234 monoclonal antibody recognized a conformation-dependent epitope. The immunoglobulin class of these 3 mAbs is determined by ELISA with a Mouse-Hybridoma Subtyping Kit (Zymed). The isotype of the anti-CPiV-HN(d94) mAb was IgM. The isotype of anti-CAV-fiber(d41) mAbs both were IgG1. Deletion mutation analysis showed that two epitopes recognized by anti-CAV-fiber(d41) mAbs were located in the amino acids residues 1-251 and 252-502 of CAV-fiber(d41) protein, respectively. The anti-CPiV-HN(d94) mAb was located in the amino acids residues 101-141 of CPiV-HN(d94) protein. In the present study, monoclonal antibodies prepared in this study will be used in development of canine multivalent vaccines and in preparation of immunochromatographic test (ICT) for rapid diagnosis of canine pathogen.