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1.
Virol J ; 19(1): 116, 2022 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-35831876

RESUMO

BACKGROUND: Bovine parainfluenza virus type 3 (BPIV3) infection often causes respiratory tissue damage and immunosuppression and further results in bovine respiratory disease complex (BRDC), one of the major diseases in dairy cattle, caused huge economical losses every year. However, the pathogenetic and immunoregulatory mechanisms involved in the process of BPIV3 infection remain unknown. However, the pathogenetic and immunoregulatory mechanisms involved in the process of BPIV3 infection remain unknown. Proteomics is a powerful tool for high-throughput identification of proteins, which has been widely used to understand how viruses interact with host cells. METHODS: In the present study, we report a proteomic analysis to investigate the whole cellular protein alterations of MDBK cells infected with BPIV3. To investigate the infection process of BPIV3 and the immune response mechanism of MDBK cells, isobaric tags for relative and absolute quantitation analysis (iTRAQ) and Q-Exactive mass spectrometry-based proteomics were performed. The differentially expressed proteins (DEPs) involved in the BPIV3 invasion process in MDBK cells were identified, annotated, and quantitated. RESULTS: A total of 116 proteins, which included 74 upregulated proteins and 42 downregulated proteins, were identified as DEPs between the BPIV3-infected and the mock-infected groups. These DEPs included corresponding proteins related to inflammatory response, immune response, and lipid metabolism. These results might provide some insights for understanding the pathogenesis of BPIV3. Fluorescent quantitative PCR and western blotting analysis showed results consistent with those of iTRAQ identification. Interestingly, the upregulated protein MKK3 was associated with the p38 MAPK signaling pathway. CONCLUSIONS: The results of proteomics analysis indicated BPIV3 infection could activate the p38 MAPK pathway to promote virus replication.


Assuntos
Vírus da Parainfluenza 3 Humana , Proteômica , Animais , Bovinos , Vírus da Parainfluenza 3 Bovina/fisiologia , Replicação Viral/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno
2.
Vet Microbiol ; 271: 109488, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35691094

RESUMO

Bovine parainfluenza virus 3 (BPIV3) is an important respiratory pathogen of both young and adult cattle. The pathways of cell entry are highly related to viral transmission and pathogenicity. In previous studies, we demonstrated that macropinocytosis and clathrin-dependent endocytosis play critical roles in the entry of BPIV3 into MDBK cells. Macropinocytosis is special endocytic process which need to activate signaling pathways that remodle the actin cytoskeleton. Parainfluenza viruses (PIVs) initiate infection by binding to sialic acid receptors on cell surfaces. Nevertheless, sialic acids are not able to transmit signals across the plasma membrane, indicating the necessity for additional signaling receptors. Here, we have demonstrated that specific inhibitors and siRNAs targeting EGFR inhibit the entry of BPIV3 into MDBK cells. BPIV3 productive infection in MDBK cells led to activation of EGFR. Inactivation of EGFR suppressed BPIV3-induced rearrangement of the F-actin cytoskeleton. In addition, PI3K-Akt and ERK1/2 were activated in an EGFR-dependent manner during BPIV3 infection. Specific inhibitors targeting these canonical downstream effectors of EGFR could significantly reduce viral entry efficacy. Moreover, we also demonstrated that the important regulators of macropinocytosis, Rac1 and Pak1, are downstream mediators of EGFR during BPIV3 internalization. These results indicated that EGFR is a host-entry cofactor used by BPIV3 to enter MDBK cells.


Assuntos
Vírus da Parainfluenza 3 Bovina , Vírus da Parainfluenza 3 Humana , Animais , Bovinos , Receptores ErbB/genética , Receptores ErbB/metabolismo , Vírus da Parainfluenza 3 Bovina/fisiologia , Fosfatidilinositol 3-Quinases , Internalização do Vírus
3.
Vet Microbiol ; 268: 109415, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35395543

RESUMO

Bovine parainfluenza virus type 3 (BPIV3) is one of the most important viral respiratory pathogens of cattle. No specific therapies are available for BPIV3 infection; vaccination is one of the most effective ways to prevent BPIV3 infection. We therefore prepared the self-assembled BPIV3 nanoparticles by genetically fusing the ectodomain of BPIV3 haemagglutinin-neuraminidase (HN) (HNex) to the NH2 terminus of ferritin (HNex-RFNp) using a baculovirus expression system. It was found that HNex-RFNp-induced bone marrow-derived dendritic cell (BMDC) maturation through the upregulated expression of surface molecules (MHC II, CD80, CD86, and CD40), increased the secretion of inflammatory cytokines (IL-6, IL-12, TNF-α, and IFN-γ), and reduced antigen phagocytosis and T cell activation capacity. HNex-RFNp positively regulated IκBα and NF-κB (p65) phosphorylation and facilitated NF-κB (p65) translocation into the nuclei of mature BMDCs. Incubating RFNp-treated BMDCs with TLR4 and NF-κB (p65) inhibitors, suppressed surface molecule expression as well as pro-inflammatory cytokine production and IκBα and NF-κB (p65) activities. The BPIV3 HNex protein induced BMDC maturation to some extent but was significantly weaker than HNex-RFNp. We found that HNex-RFNp induced a higher titre of specific antibodie, haemagglutinin inhibition (HI) antibody, and virus neutralisation (VN) antibody, and a comprehensive cellular immune response. We examined protection against BPIV3 challenge in a mouse model. Pathological changes were not observed in the lungs of HNex-RFNp-vaccinated mice. Levels of BPIV3 RNA and virus titres in the lungs and trachea were significantly lower in the HNex-RFNp, than HNex, inactivated BPIV3, and PBS groups. In summary, HNex-RFNp elicited better immunogenicity than HNex or inactivated BPIV3 and could be developed as an effective vaccine to protect against BPIV3 infection.


Assuntos
Células Dendríticas , NF-kappa B , Nanopartículas , Vírus da Parainfluenza 3 Bovina , Vacinas Virais , Viroses , Animais , Bovinos , Células Dendríticas/imunologia , Hemaglutininas/metabolismo , Imunogenicidade da Vacina , Ativação Linfocitária , Camundongos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Vacinas Virais/imunologia , Viroses/prevenção & controle , Viroses/veterinária
4.
J Dairy Sci ; 105(1): 560-571, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34763911

RESUMO

The role of colostrum management in providing adequate immunological protection to neonatal calves has been widely investigated, and thresholds for colostrum quality, as well as optimum volume and timing for colostrum feeding have been established. However, limited information is available on the effect of colostrum source (single dam or pooled) on passive immunity, as well as subsequent antibody survival in the calf. This study aimed to assess the effect of feeding single-dam colostrum (own and other dam) or pooled colostrum on transfer of passive immunity, and also investigate the rate of depletion of disease-specific antibodies among dairy calves. In total, 320 cows and 119 dairy heifer calves were enrolled in the study. Calves were blood-sampled immediately after birth and received either own-dam, other-dam, or pooled colostrum. Calves were blood-sampled at 24 h to assess serum IgG concentrations and at monthly intervals thereafter to document disease-specific antibody survival. Mean colostrum IgG concentration was higher for other-dam treatment group, whereas own-dam and pooled treatments were similar. For all treatment groups, the mean IgG concentration was >80 mg/mL, exceeding the quality threshold of 50 mg/mL. Mean calf serum IgG concentration was lower for calves fed pooled colostrum compared with those that received colostrum from a single cow. There was a negative association with 24-h serum IgG and calf birth bodyweight; calves <30 kg at birth had the highest 24-h serum IgG concentration. Survival of antibodies to bovine viral diarrhea, Salmonella infection, leptospirosis, bovine parainfluenza 3 virus, bovine respiratory syncytical virus, rotavirus, and coronavirus was not associated with colostrum source; however, antibodies to infectious bovine rhinotracheitis had a greater period of survival among calves fed own-dam colostrum. We found that feeding single-dam colostrum can thus improve calf immunity through increased serum IgG levels and antibody survival rates. Furthermore, we hypothesize that immune exclusion may occur with pooled colostrum; therefore, providing pooled colostrum may still be a good practice as long as it can be ensured that enough antibodies are absorbed into the blood stream to deal with pathogens calves may encounter because different dams may have antibodies against different strains of viruses and bacteria, yielding cross protection.


Assuntos
Colostro , Vírus da Parainfluenza 3 Bovina , Animais , Animais Recém-Nascidos , Bovinos , Feminino , Imunidade Materno-Adquirida , Imunoglobulinas , Parto , Gravidez
5.
Proc Natl Acad Sci U S A ; 118(50)2021 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-34876520

RESUMO

Single-dose vaccines with the ability to restrict SARS-CoV-2 replication in the respiratory tract are needed for all age groups, aiding efforts toward control of COVID-19. We developed a live intranasal vector vaccine for infants and children against COVID-19 based on replication-competent chimeric bovine/human parainfluenza virus type 3 (B/HPIV3) that express the native (S) or prefusion-stabilized (S-2P) SARS-CoV-2 S spike protein, the major protective and neutralization antigen of SARS-CoV-2. B/HPIV3/S and B/HPIV3/S-2P replicated as efficiently as B/HPIV3 in vitro and stably expressed SARS-CoV-2 S. Prefusion stabilization increased S expression by B/HPIV3 in vitro. In hamsters, a single intranasal dose of B/HPIV3/S-2P induced significantly higher titers compared to B/HPIV3/S of serum SARS-CoV-2-neutralizing antibodies (12-fold higher), serum IgA and IgG to SARS-CoV-2 S protein (5-fold and 13-fold), and IgG to the receptor binding domain (10-fold). Antibodies exhibited broad neutralizing activity against SARS-CoV-2 of lineages A, B.1.1.7, and B.1.351. Four weeks after immunization, hamsters were challenged intranasally with 104.5 50% tissue-culture infectious-dose (TCID50) of SARS-CoV-2. In B/HPIV3 empty vector-immunized hamsters, SARS-CoV-2 replicated to mean titers of 106.6 TCID50/g in lungs and 107 TCID50/g in nasal tissues and induced moderate weight loss. In B/HPIV3/S-immunized hamsters, SARS-CoV-2 challenge virus was reduced 20-fold in nasal tissues and undetectable in lungs. In B/HPIV3/S-2P-immunized hamsters, infectious challenge virus was undetectable in nasal tissues and lungs; B/HPIV3/S and B/HPIV3/S-2P completely protected against weight loss after SARS-CoV-2 challenge. B/HPIV3/S-2P is a promising vaccine candidate to protect infants and young children against HPIV3 and SARS-CoV-2.


Assuntos
Vacinas contra COVID-19/administração & dosagem , COVID-19/prevenção & controle , SARS-CoV-2/imunologia , Administração Intranasal , Animais , Anticorpos Antivirais/sangue , Vacinas contra COVID-19/genética , Vacinas contra COVID-19/imunologia , Cricetinae , Vetores Genéticos , Imunização , Vírus da Parainfluenza 3 Bovina/genética , Vírus da Parainfluenza 3 Humana/genética , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/imunologia , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
6.
Vet Microbiol ; 261: 109185, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34364015

RESUMO

Bovine parainfluenza-3 virus (BPIV-3) is one of the main viruses associated with bovine respiratory disease complex (BRDC) worldwide. BPIV-3 infect the bovine respiratory tract causing from subclinical infections to severe pneumonia with significant economic losses in the cattle industry. BPIV-3 is a RNA virus with high genetic variability, nevertheless, the contribution of recombination events to its variability has not been assessed so far. In this study the 25 complete genome sequences (CGS) reported so far and 215 partial sequences of different viral genes of BPIV-3 were analyzed to determine their genotypes and subgenotypes, distribution, and the existence of potential recombination events. Based on the analysis of the HN, M, N, and P genes one hypothetical subgenotype was found (subgenotype A4). Four recombination events between sequences of swine and cattle were detected by RDP4 analysis in conjunction with phylogenetic incongruences in the L gene. In addition, 9 sequences reported from Argentina were found to be miss-classified. These results reveal that homologous recombination events have a relevant role in the evolution of BPIV-3 and highlight the importance of implement advanced molecular characterization to better understand the variability and evolution of BPIV-3 as a component of BRDC.


Assuntos
Variação Genética/genética , Recombinação Homóloga/genética , Vírus da Parainfluenza 3 Bovina/genética , Proteínas Virais/genética , Animais , Bovinos , Doenças dos Bovinos/virologia , Genótipo , Vírus da Parainfluenza 3 Bovina/classificação , Filogenia , Infecções por Respirovirus/virologia , Ovinos , Doenças dos Ovinos/virologia
7.
Viruses ; 13(6)2021 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-34072688

RESUMO

Bovine parainfluenza virus 3 (BPIV3) is a crucial causative agent of respiratory disease in young and adult cattle. No specific therapies are available for BPIV3 infection. Understanding the internalization pathway of the virus will provide a new strategy for the development of antiviral therapy. Here, the mechanism of BPIV3 entry into HeLa cells was analyzed using RNA silencing and pharmacological inhibitors. Treatment of HeLa cells with hypertonic medium prevented BPIV3 internalization. These results indicated that BPIV3 entered HeLa cells via receptor-mediated endocytosis. Moreover, removing cell membrane cholesterol through MßCD treatment hampered viral penetration but not viral replication. In addition, BPIV3 infection was inhibited by pretreatment with dynasore or chlorpromazine (CPZ) or knockdown of dynamin II or clathrin heavy chain. However, virus entry was unaffected by nystatin, EIPA, wortmannin, or cytochalasin D treatment or caveolin-1 knockdown. These data demonstrated that the entry of BPIV3 into HeLa cells was dependent on clathrin-mediated endocytosis but not on caveolae-mediated endocytosis or the macropinocytosis pathway. Many viruses are transported to endosomes, which provide an acidic environment and release their genome upon separation from primary endocytic vesicles. However, we found that BPIV3 infection required endosomal cathepsins, but not a low pH. In summary, we show, for the first time, that BPIV3 enters HeLa cells through the clathrin-mediated endocytosis pathway, presenting novel insights into the invasion mechanism of Paramyxoviridae.


Assuntos
Colesterol/metabolismo , Clatrina/metabolismo , Dinaminas/metabolismo , Endocitose , Vírus da Parainfluenza 3 Bovina/fisiologia , Internalização do Vírus , Animais , Bovinos , Células HeLa , Humanos
8.
Virology ; 561: 17-27, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34130198

RESUMO

Paramyxovirus matrix (M) proteins are key drivers of virus particle assembly and budding at the plasma membrane. To identify regions important for the M protein function, we generated a series of deletion mutants of the bovine parainfluenza virus type 3 (BPIV3) M protein. We found that M proteins lacking 10 amino acids in the amino-terminal end (ΔN10) or 4 amino acids in the carboxyl-terminal end (ΔC4) did not support M-deficient BPIV3 virion release and M protein-induced virus-like particle (VLP) release. Both ΔN10 and ΔC4 retained M protein-M protein and M protein-nucleocapsid (N) protein interactions. However, neither was transported to the plasma membrane. Our results indicate that both amino- and carboxyl-terminal ends of the BPIV3 M protein are essential for M protein transport to the plasma membrane, where it facilitates virion and VLP release.


Assuntos
Vírus da Parainfluenza 3 Bovina/fisiologia , Proteínas da Matriz Viral/química , Proteínas da Matriz Viral/metabolismo , Vírion/fisiologia , Liberação de Vírus , Animais , Membrana Celular/metabolismo , Chlorocebus aethiops , Proteínas Mutantes/metabolismo , Proteínas do Nucleocapsídeo/metabolismo , Vírus da Parainfluenza 3 Bovina/química , Transporte Proteico , Deleção de Sequência , Células Vero , Proteínas da Matriz Viral/genética
9.
Vet Microbiol ; 259: 109148, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34147763

RESUMO

Bovine parainfluenza virus 3 (BPIV3) is an important respiratory pathogen of both young and adult cattle. No specific therapies are available for BPIV3. Understanding the viral internalization pathway of BPIV3 will provide new strategies for the development of antiviral treatments. Here, the entry mechanism of BPIV3 into MDBK cells was analyzed using chemical inhibitors and RNA silencing. Our data demonstrated that treatment with an inhibitor targeting the clathrin-mediated pathway or clathrin heavy chain (CHC) knockdown suppressed the entry of BPIV3 into MDBK cells. In contrast, sequestration of cellular cholesterol by nystatin or silencing of caveolin-1 had no effect on viral entry. Moreover, inhibition of critical modulators of macropinocytosis significantly reduced BPIV3 uptake. In addition, fluid-phase uptake was significantly increased in cells infected with BPIV3, which is indicative of virus-induced facilitation of macropinocytosis. These results suggest that BPIV3 enters MDBK cells via macropinocytosis and clathrin- but not caveolar-dependent endocytosis. Furthermore, inhibition of endosomal acidification and activation of cathepsin blocked BPIV3 entry, demonstrating that BPIV3 entered MDBK cells in a acid-dependent manner and required cathepsin L. Finally, we demonstrated that macropinocytosis but not clathrin-mediated endocytosis is dependent on actin dynamics during BPIV3 infection.


Assuntos
Ácidos/metabolismo , Cadeias Pesadas de Clatrina/genética , Clatrina/metabolismo , Endocitose , Vírus da Parainfluenza 3 Bovina/fisiologia , Pinocitose , Internalização do Vírus , Animais , Catepsina L/metabolismo , Bovinos , Linhagem Celular , Cadeias Pesadas de Clatrina/metabolismo
10.
Vet Med Sci ; 7(5): 1625-1632, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34031994

RESUMO

Bovine parainfluenza virus-3 (BPIV-3), also known as bovine respirovirus 3, causes serious respiratory infection in ungulates, often involving other pathogens, such as viruses, bacteria and mycoplasmas. In this study, we evaluated antibody titers against virus genotypes A (BPIV-3a) and C (BPIV-3c). We conducted a serological survey and comparison analysis of archived serum samples from small and large ruminants reared in four Turkish provinces. A total of 1,307 samples, consisting of sheep (n = 444), cattle (n = 402), water buffalo (n = 261) and goat (n = 200) sera, were randomly selected from stock samples collected between 2015 and 2019 and screened by standard virus neutralisation assay. We found that 49.9% (653/1307) of all samples were positive for neutralising antibody titers. Goats had the highest titer, with total seropositivity of 63% (126/200), followed in descending order by cattle, sheep and water buffalo at 56.2% (226/402), 32.2% (143/444) and 26% (68/261) total seropositivity, respectively. BPIV-3c had the highest neutralising antibody rate at 34.3% (448/1307), whereas BPIV-3a had a 24.3% (317/1307) seropositivity rate. Neutralising antibody titers for positive samples ranged between 1/4 and 1/512 per the SN50 test. Seropositivity rates ranged from a low of 8.9% to a high of 18.3%. Our study was the first to compare antibody seroprevalence for two BPIV-3 genotypes in small and large domestic ruminants, which were shown to be more commonly exposed to BPIV-3c than BPIV-3a. This finding could have significant implications as current vaccines mainly use the BPIV-3a genotype. Further research can determine if current vaccines protect against different BPIV-3 virus genotypes.


Assuntos
Cabras , Vírus da Parainfluenza 3 Bovina , Animais , Búfalos , Bovinos , Genótipo , Vírus da Parainfluenza 3 Bovina/genética , Estudos Soroepidemiológicos , Ovinos
11.
Can J Vet Res ; 85(2): 101-105, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33883816

RESUMO

Abruptly weaned crossbred steer calves (N = 271) were used in a randomized, blinded 2-arm clinical trial to assess the impact of a long-acting non-steroidal anti-inflammatory drug on bovine herpesvirus type 1, bovine respiratory syncytial virus, parainfluenza virus type 3, and coronavirus titers and health outcomes when administered concurrently with a modified live respiratory vaccine upon arrival at a feedlot. Treatment groups included a control (saline; n = 135) and an experimental group (injectable meloxicam; n = 136). Viral antibody titers and body weight were measured on arrival, day 7, and day 21, along with a final weight on day 45. Body weight and antibody titers for all viruses increased over time (P < 0.001); however, there were no differences by treatment group or a significant group × time interaction when evaluated using repeated measures analysis of variance. Interestingly, the use of meloxicam was associated with increased treatment risk (P < 0.05). In conclusion, the administration of meloxicam may adversely affect health; however, a decreased vaccine response is likely not a contributing factor.


Des bouvillons croisés sevrés rapidement (N = 271) ont été utilisés dans un essai clinique randomisé en aveugle à deux bras pour évaluer l'impact d'un anti-inflammatoire non stéroïdien à action prolongée sur les titres du virus de la rhinotrachéite infectieuse bovine, du virus respiratoire syncytial bovin, du virus parainfluenza 3 et du coronavirus, et les résultats pour la santé lorsqu'administré en même temps qu'un vaccin vivant modifié respiratoire à l'arrivée dans un parc d'engraissement. Les groupes de traitement comprenaient un témoin (solution saline; n = 135) et un groupe expérimental (méloxicam injectable; n = 136). Les titres d'anticorps viraux et le poids corporel ont été mesurés à l'arrivée, au jour 7 et au jour 21, ainsi qu'un poids final au jour 45. Le poids corporel et les titres d'anticorps pour tous les virus ont augmenté avec le temps (P < 0,001); cependant, il n'y avait aucune différence selon le groupe de traitement ou une interaction groupe × temps significative lors de l'évaluation à l'aide de mesures répétées d'analyse de la variance. Fait intéressant, l'utilisation du méloxicam était associée à un risque de traitement accru (P < 0,05). En conclusion, l'administration de méloxicam peut nuire à la santé; cependant, une réponse vaccinale réduite n'est probablement pas un facteur contributif.(Traduit par Docteur Serge Messier).


Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Meloxicam/administração & dosagem , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Anticorpos Antivirais/sangue , Bovinos , Coronavirus Bovino/imunologia , Herpesvirus Bovino 1/imunologia , Masculino , Meloxicam/farmacologia , Meloxicam/uso terapêutico , Vírus da Parainfluenza 3 Bovina/imunologia , Vírus Sincicial Respiratório Bovino/imunologia , Desmame
12.
Vet Q ; 41(1): 97-106, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33375918

RESUMO

BACKGROUND: The knowledge on bovine vaccines against respiratory viruses on bronchoalveolar fluid cells is scarce. OBJECTIVE: To compare the effects of a commercial intranasal (IN) and intramuscular (IM) vaccine against bovine respiratory disease (BRD) complex viruses on bronchoalveolar fluid cells of healthy heifers. METHODS: 21 healthy heifers were assigned to three treatment groups: control (CO, N = 7), intranasally vaccinated (IN) (n = 7), and intramuscularly vaccinated (IM) (n = 7). The IN group received 1 mL of the commercial vaccine in each nostril once containing attenuated BoHV-1, bPIV-3, and BRSV. The IM group was vaccinated with two doses of 2 mL with an interval of 21 days of the commercial vaccine containing attenuated BoHV-1, bPIV-3, and BRSV plus inactivated BVDV. At day 0 (D0), before the first vaccine dose, and at D3, D7, and D21, after the last vaccine dose, airway bronchoscopy was performed to observe local irritation and collect bronchoalveolar lavage fluid (BALF). The bronchoalveolar count, cytological evaluation, bronchoalveolar cell oxidative metabolism, and total bronchoalveolar IgA and IgG were measured. RESULTS: The IN vaccine increased neutrophil cellularity at D7 and D21 and total IgA at D3 in BALF. Total IgA in BALF also increased at D3 and oxidative metabolism of bronchoalveolar cells at D21 lowered compared to the CO group. Following IM vaccination there was no alteration of immunoglobulins or cell oxidative metabolism in BALF. Both vaccines reduced the number of alveolar macrophages. CONCLUSION: Both vaccines induced bronchoalveolar inflammation during the establishment of the vaccine immunity, which was more expressive in the IN protocol.


Assuntos
Líquido da Lavagem Broncoalveolar/citologia , Doenças dos Bovinos/prevenção & controle , Vacinação/veterinária , Administração Intranasal/efeitos adversos , Administração Intranasal/veterinária , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/prevenção & controle , Líquido da Lavagem Broncoalveolar/química , Bovinos , Doenças dos Bovinos/virologia , Vírus da Diarreia Viral Bovina , Feminino , Infecções por Herpesviridae/prevenção & controle , Herpesvirus Bovino 1 , Imunoglobulina A , Imunoglobulina G , Injeções Intramusculares/efeitos adversos , Injeções Intramusculares/veterinária , Vírus da Parainfluenza 3 Bovina , Infecções por Vírus Respiratório Sincicial/prevenção & controle , Infecções por Vírus Respiratório Sincicial/veterinária , Vírus Sincicial Respiratório Bovino , Infecções por Respirovirus/prevenção & controle , Infecções por Respirovirus/veterinária , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/efeitos adversos , Vacinas Virais/administração & dosagem
13.
Vet Immunol Immunopathol ; 230: 110130, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33068827

RESUMO

Almost all bovine intranasal respiratory vaccines require a spraying device, but there are no published reports which prove the necessity of this. The objective of this investigation was to compare the efficacy of the Bovilis® INtranasal RSP™ Live vaccine when applied with or without an (intra)nasal spraying device. This vaccine contains live, attenuated strains of bovine respiratory syncytial virus (BRSV) and bovine parainfluenza virus type 3 (BPI3V) and is licensed to protect young cattle against respiratory infections with wild-type field isolates of these viruses. Two efficacy studies, for BRSV and BPI3V respectively, were performed in which the vaccine was administered using a spraying device or directly from the tip of a syringe as a liquid stream. Both BRSV-vaccinated groups showed a reduction of nasal shedding, BRSV titres in lung washings and clinical symptoms. The BPI3V vaccinated groups showed a significant reduction of nasal shedding and a non-significant reduction of clinical symptoms. Overall, in both studies, the groups in which vaccine was administered without the spraying device performed better than the groups with the spraying device, although this difference was not statistically significant. In conclusion, a spraying device to administer Bovilis® INtranasal RSP™ Live was not required and both application methods induced a protective immune response. This makes application more convenient and flexible for future users and animals.


Assuntos
Administração Intranasal/instrumentação , Administração Intranasal/veterinária , Infecções Respiratórias/prevenção & controle , Infecções Respiratórias/veterinária , Vacinação/métodos , Vacinação/veterinária , Vacinas Virais/administração & dosagem , Aerossóis/administração & dosagem , Fatores Etários , Animais , Anticorpos Antivirais/sangue , Bovinos , Doenças dos Bovinos/prevenção & controle , Sprays Nasais , Vírus da Parainfluenza 3 Bovina/imunologia , Vírus Sincicial Respiratório Bovino/imunologia , Vacinação/instrumentação , Vacinação/normas , Vacinas Atenuadas/administração & dosagem
14.
Vet J ; 263: 105532, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32928493

RESUMO

Bovine respiratory syncytial virus (BRSV) and bovine parainfluenza-3 virus (bPI3V) are major causes of bovine respiratory disease (BRD) in newborn calves worldwide. Vaccination is widely used to prevent BRD, and intranasal vaccines for BRSV and bPI3V were developed to overcome interference from BRSV and bPI3V-specific maternally derived antibodies. Many experimental challenge trials have demonstrated that intranasal vaccines for BRSV and bPI3V are efficacious, but effectiveness under field conditions has been demonstrated less often, especially for newborn beef calves. The objective of this field trial was to compare the effectiveness of a newly available commercial BRSV-bPI3V intranasal vaccine with that of a benchmarked one in newborn beef calves reared in a cow-calf system. A total of 935 calves from 39 farms were randomized into two vaccine groups (Bovalto Respi Intranasal [Vaccine A], n=468; Rispoval RS+PI3 Intranasal [Vaccine B], n=467), and monitored during the in-house risk period up to three months after vaccination. Non-inferiority analysis was performed by calculating the difference in BRD prevalence between the two vaccine groups. No significant differences were observed between vaccines regarding clinical outcomes of morbidity, mortality, duration between vaccination and BRD occurrence, or treatments required. Because the upper limit of the 2-sided 95% confidence interval of the difference in BRD prevalence between the two treatment groups (0.8%) was less than the margin of non-inferiority (δ=5%), a non-inferiority of Vaccine A was concluded. In conclusion, Vaccine A is at least as effective as Vaccine B for the prevention of BRD in newborn beef cattle in a cow-calf system under field conditions.


Assuntos
Animais Recém-Nascidos , Doenças dos Bovinos/prevenção & controle , Vírus da Parainfluenza 3 Bovina/imunologia , Vírus Sincicial Respiratório Bovino/imunologia , Infecções Respiratórias/veterinária , Vacinas Virais/administração & dosagem , Administração Intranasal/veterinária , Animais , Bovinos , Feminino , Masculino , Infecções por Vírus Respiratório Sincicial/prevenção & controle , Infecções por Vírus Respiratório Sincicial/veterinária , Infecções Respiratórias/prevenção & controle , Infecções Respiratórias/virologia , Infecções por Respirovirus/prevenção & controle , Infecções por Respirovirus/veterinária , Resultado do Tratamento
15.
Vet Microbiol ; 247: 108774, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32768220

RESUMO

Bovine parainfluenza virus type 3 (BPIV3) is one of the most important viral respiratory pathogens of cattle. In addition to the classical BPIV3 genotype A (BPIV3a), new genetic groups, genotype B (BPIV3b) and C (BPIV3c), have been identified and isolated in certain parts of the world. The present study aimed to investigate the genetic and antigenic characteristics of BPIV3 circulating in Japan. Seventy-three BPIV3 field strains were isolated from nasal samples of cattle between 2002 and 2019. Phylogenetic analysis of the phosphoprotein and hemagglutinin-neuraminidase genes showed that the isolates clustered into two genotypes, BPIV3a (49 %) and BPIV3c (51 %). The BPIV3a strains had more wide genetic variation than the rest of the genotypes. Additionally, new variants were obtained and designated them tentatively as subgroup 4 of the BPIV3a. The first Japanese BPIV3c was isolated in 2012, but here the BPIV3c NM2 strain was isolated from a sample collected four years earlier than the previous report. The antigenicity of ten BPIV3 strains including all three genotypes was assessed with a viral cross-neutralization test. Anti-sera against BPIV3a and BPIV3b cross-reacted well with both homologous and heterologous viruses. On the other hand, anti-sera against BPIV3c had reduced cross-reactivity to the heterologous viruses. Overall, our findings showed that genetically and antigenically divergent BPIV3 is prevalent in cattle in Japan. These results could provide a reference for molecular epidemiological characterization of BPIV3 and vaccine development.


Assuntos
Antígenos Virais/genética , Vírus da Parainfluenza 3 Bovina/classificação , Vírus da Parainfluenza 3 Bovina/imunologia , Filogenia , Infecções por Respirovirus/epidemiologia , Infecções por Respirovirus/veterinária , Animais , Bovinos/virologia , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Indústria de Laticínios , Feminino , Genótipo , Japão/epidemiologia , Masculino , Nariz/virologia , Prevalência
16.
Can J Vet Res ; 84(3): 163-171, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32801450

RESUMO

Bovine respiratory disease complex is etiologically complex and usually involves co-infection by several agents, including bovine parainfluenza virus-3 (BPIV-3), bovine respiratory syncytial virus (BRSV), and bovine coronavirus (BCoV). Traditionally, vaccines have been tested in seronegative calves infected with a single in vitro-passaged agent, often with little disease, resulting in unvaccinated subjects. To overcome the potential problem of attenuation coincident with in vitro culture of the viruses, cocktails of field isolates of BPIV-3s and BCoVs were passaged in the lungs of neonatal colostrum-deprived calves. Lung lavage fluids were used as inocula, alone and in combination with in-vivo passaged BRSV, and aerosolized into a trailer containing conventionally reared 9-week-old weaned Holstein calves with decayed, but still measurable, maternal antibodies. Calves developed acute respiratory disease of variable severity. Upon necropsy, there were characteristic gross and histologic lesions in the respiratory tract, associated immunohistochemically with BPIV-3, BRSV, and BCoV. In-vivo passage of viruses is an alternative to in vitro culture to produce inocula to better study the pathogenesis of infection and more rigorously and relevantly assess vaccine efficacy.


Le complexe des maladies respiratoires bovines possède une étiologie complexe et implique habituellement une co-infection par plusieurs agents, incluant le virus parainfluenza bovin 3 (BPIV-3), le virus respiratoire syncitial bovin (BRSV) et le coronavirus bovin (BCoV). Traditionnellement, les vaccins ont été testés chez des veaux séronégatifs infectés avec un seul agent cultivé in vitro, présentant souvent peu de maladie, résultant en des sujets non-vaccinés. Afin de contrecarrer le problème potentiel d'atténuation associé à la culture in vitro des virus, des cocktails d'isolats de champs de BPIV-3 et de BCoV furent passés dans des poumons de veaux nouveau-nés privés de colostrum. Les liquides de lavage pulmonaire furent utilisés comme inoculum, seul et en combinaison avec des BRSV passés in vivo, et aérosolisés dans une remorque contenant des veaux Holstein sevrés élevés de manière conventionnelle âgés de 9 semaines ayant des anticorps maternels en déclin mais toujours mesurables. Les veaux ont développé une maladie respiratoire aiguë de sévérité variable. Lors de la nécropsie, il y avait des lésions macroscopiques et histologiques caractéristiques dans le tractus respiratoire, associées immuno-histochimiquement avec BPIV-3, BRSV et BCoV. Le passage in vivo de virus est une alternative à la culture in vitro afin de produire un inoculum permettant de mieux étudier la pathogénie de l'infection et d'évaluer plus rigoureusement et plus pertinemment l'efficacité de vaccins.(Traduit par Docteur Serge Messier).


Assuntos
Doenças dos Bovinos/virologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/patogenicidade , Vírus da Parainfluenza 3 Bovina/patogenicidade , Infecções por Vírus Respiratório Sincicial/veterinária , Infecções por Respirovirus/veterinária , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/isolamento & purificação , Bovinos , Doenças dos Bovinos/patologia , Infecções por Coronavirus/complicações , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Coronavirus Bovino/isolamento & purificação , Imuno-Histoquímica/veterinária , Pulmão/patologia , Pulmão/virologia , Vírus da Parainfluenza 3 Bovina/imunologia , Vírus da Parainfluenza 3 Bovina/isolamento & purificação , Atelectasia Pulmonar/patologia , Atelectasia Pulmonar/veterinária , Infecções por Vírus Respiratório Sincicial/complicações , Infecções por Vírus Respiratório Sincicial/patologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Vírus Sinciciais Respiratórios/patogenicidade , Infecções por Respirovirus/complicações , Infecções por Respirovirus/patologia , Infecções por Respirovirus/virologia , Traqueia/patologia , Traqueia/virologia
17.
BMC Vet Res ; 16(1): 72, 2020 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-32127006

RESUMO

BACKGROUND: Bovine parainfluenza virus type 3 (BPIV3) is one of the important viral respiratory agents associated with the bovine respiratory disease complex (BRDC) in cattle. Previous study has demonstrated that infection of BPIV3 causes innate immune response within the host cell. ß-catenin is a key component of the Wnt/ß-catenin signal pathway which is involved in the regulation of interferon-beta (IFN-ß) transcription. Some viruses can activate while others can inhibit the Wnt/ß-catenin signaling pathway. However, the role of ß-catenin in BPIV3 infection remains unclear. RESULTS: Here we found that the expression of ß-catenin mRNA was up-regulated and ß-catenin protein was down-regulated after BPIV3 infection in MDBK cells. Moreover, it was confirmed that overexpression of ß-catenin suppressed BPIV3 replication and knockdown of ß-catenin promoted viral replication, suggesting that ß-catenin inhibits BPIV3 replication. Furthermore, IFN-ß signal pathway and virus titer analysis using the GSK3ß inhibitor (LiCl) revealed that Wnt/ß-catenin can serve as a mechanism to suppress virus replication in infected cells. The results indicated that LiCl promoted the expression and accumulation in the nucleus of ß-catenin, which further promoted the expression of IFN-ß and OSA1 and suppressed BPIV3 replication. Most importantly, BPIV3 down-regulating ß-catenin protein expression was due to degradation of GSK3ß mediated proteasome pathway. CONCLUSIONS: In summary, we discovered the relationship between ß-catenin and BPIV3 replication. These results provided further insight into the study of BPIV3 pathogenesis.


Assuntos
Imunidade Inata , Vírus da Parainfluenza 3 Bovina/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , beta Catenina/metabolismo , Animais , Bovinos , Linhagem Celular , Glicogênio Sintase Quinase 3 beta/efeitos dos fármacos , Cloreto de Lítio/farmacologia , RNA Mensageiro , Infecções por Respirovirus/imunologia , Infecções por Respirovirus/veterinária , Transdução de Sinais , beta Catenina/genética
18.
J Dairy Sci ; 103(3): 2556-2566, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31954585

RESUMO

Respiratory tract infections (bovine respiratory disease) are a major concern in calf rearing. The objective of this study was to identify pathogen-specific risk factors associated with epidemic respiratory disease in calves. A cross-sectional study was conducted, involving 128 outbreaks (29 dairy, 58 dairy-mixed, and 41 beef) in Belgium (2016-2018). A semiquantitative PCR for 7 respiratory pathogens was done on a pooled nonendoscopic bronchoalveolar lavage sample for each herd. Potential risk factors were collected by questionnaire and derived from the national cattle registration databank. Most outbreaks occurred between October and March, and single and multiple viral infections were detected in 58.6% (75/128) and 13.3% (17/128), respectively. Bovine coronavirus (BCV) was the most frequently isolated virus (38.4%), followed by bovine respiratory syncytial virus (bRSV; 29.4%) and parainfluenzavirus type 3 (PI-3; 8.1%). Mycoplasma bovis, Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni were detected in 33.3, 41.2, 89.1, and 36.4% of the herds, respectively. Specific risk factors for BCV detection were detection of M. haemolytica [odds ratio (OR) = 2.8 (95% confidence interval = 1.1-7.5)], increasing herd size [OR = 1.3 (1.0-1.8) for each increase with 100 animals] and detection of BCV by antigen ELISA on feces in calves in the last year [OR = 3.6 (1.2-11.1)]. A seasonal effect was shown for bRSV only {more in winter compared with autumn [OR = 10.3 (2.8-37.5)]}. Other factors associated with bRSV were PI-3 detection [OR = 13.4 (2.1-86.0)], prevalence of calves with respiratory disease [OR = 1.02 (1.00-1.04) per 1% increase], and number of days with respiratory signs before sampling [OR = 0.99 (0.98-0.99) per day increase]. Next to its association with BCV, M. haemolytica was more frequently detected in herds with 5 to 10 animals per pen [OR = 8.0 (1.4-46.9)] compared with <5 animals, and in herds with sawdust as bedding [OR = 18.3 (1.8-191.6)]. Also, for H. somni, housing on sawdust was a risk factor [OR = 5.2 (1.2-23.0)]. Purchase of cattle [OR = 2.9 (1.0-8.0)] and housing of recently purchased animals in the same airspace [OR = 5.0 (1.5-16.5)] were risk factors for M. bovis. This study identified pathogen-specific risk factors that might be useful for the development of customized control and prevention and for the design of decision support tools to justify antimicrobial use by predicting the most likely pathogen before sampling results are available.


Assuntos
Doenças dos Bovinos/epidemiologia , Coronavirus Bovino/isolamento & purificação , Surtos de Doenças/veterinária , Infecções Respiratórias/veterinária , Animais , Bélgica/epidemiologia , Lavagem Broncoalveolar/veterinária , Bovinos , Doenças dos Bovinos/microbiologia , Estudos Transversais , Fezes/microbiologia , Feminino , Masculino , Mannheimia haemolytica/isolamento & purificação , Mycoplasma bovis/isolamento & purificação , Vírus da Parainfluenza 3 Bovina/isolamento & purificação , Pasteurella multocida/isolamento & purificação , Pasteurellaceae/isolamento & purificação , Vírus Sincicial Respiratório Bovino/isolamento & purificação , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/microbiologia , Fatores de Risco , Especificidade da Espécie , Inquéritos e Questionários
19.
Transbound Emerg Dis ; 67 Suppl 2: 82-93, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31232526

RESUMO

The etiology and pathologic findings of bovine respiratory disease (BRD) in adult dairy cows (n = 35) from a commercial dairy herd in Southern Brazil were investigated. Pulmonary samples were examined for histopathologic patterns and specific features within these patterns, while immunohistochemical (IHC) assays were designed to detect the intralesional antigens of viral infectious disease agents and Mycoplasma bovis. Pneumonia was diagnosed in 91.4% (32/35) of these cases; neither pneumonia nor any of the infectious disease pathogens evaluated occurred in three cows. The presence of multiple respiratory pathogens in 75% (24/32) of these cases indicated the complex origin of pneumonia in cattle. Interstitial pneumonia, necrosuppurative bronchopneumonia and suppurative bronchopneumonia were the principal patterns of pulmonary disease identified by histopathology. The most frequent pathogens identified by IHC were bovine viral diarrhea virus (BVDV; n = 18), M. bovis (n = 16) and bovine alphaherpesvirus type 1 (BoHV-1; n = 14), followed by bovine respiratory syncytial virus (BRSV; n = 11) and bovine parainfluenza virus type 3 (BPIV-3; n = 5). Obliterative bronchiolitis and peribronchial lymphocytic cuffings were the characteristic histopathologic features associated with M. bovis. Necrohemorrhagic bronchitis with bronchial angiogenesis was associated with BoHV-1. Necrotizing bronchitis and bronchiolitis were associated with BVDV, BoHV-1 and BRSV. Ballooning degeneration of the bronchial and bronchiolar epithelia was associated with BRSV and BoHV-1. This is the first report from Brazil that correlated the histopathologic findings of BRD with the associated infectious disease agents by immunohistochemistry. M. bovis was frequently detected in the tissues of cows with fatal pulmonary disease during this study and may be a possible primary disease pathogen associated with the development of BRD in dairy cows. Additionally, the histopathologic features identified within patterns of pulmonary disease during this investigation may be an efficient diagnostic tool to associate histopathologic findings with specific agents of BRD in dairy cows.


Assuntos
Complexo Respiratório Bovino/virologia , Herpesvirus Bovino 1/isolamento & purificação , Infecções por Mycoplasma/microbiologia , Mycoplasma bovis/isolamento & purificação , Vírus da Parainfluenza 3 Bovina/isolamento & purificação , Vírus Sincicial Respiratório Bovino/isolamento & purificação , Animais , Anticorpos Antivirais/sangue , Complexo Respiratório Bovino/diagnóstico , Brasil , Bovinos , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Herpesvirus Bovino 1/imunologia , Infecções por Mycoplasma/diagnóstico , Vírus da Parainfluenza 3 Bovina/imunologia , Transtornos Respiratórios/veterinária , Vírus Sincicial Respiratório Bovino/imunologia
20.
Pol J Vet Sci ; 23(4): 481-489, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33480488

RESUMO

Bovine parvovirus (BPV), bovine coronavirus (BCoV) and bovine parainfluenza virus (BPIV) are common etiologies causing gastrointestinal and respiratory diseases in dairy herds. However, there are few reports on the synchronous detection of BPV, BCoV and BPIV. The present article aimed to develop a quick and accurate RT-PCR assay to synchronously detect BPV, BCoV and BPIV based on their specific probes. One pair universal primers, one pair specific primers and one specific probe was designed and synthesized. After the concentrations of primer and probe and annealing temperature were strictly optimized, the specificity, sensitivity and repeatability of the established triplex probe qRT-PCR were evaluated, respectively. The results showed the recombinant plasmids of pMD18-T-BPV, pMD18-T-BCoV and pMD18-T-BPIV were 554bp, 699bp and 704bp, respectively. The optimal annealing temperature was set at 45.0°C for triplex qRT-PCR. The triplex probe qRT-PCR can only synchronously detect BPV, BCoV and BPIV. Detection sensitivities were 2.0×102, 2.0×102 and 2.0×101 copies/µL for BPV, BCoV and BPIV, being 1000-fold greater than that in the conventional PCR. Detection of clinical samples demonstrated that triplex probe qRT-PCR had a higher sensitivity and specificity. The intra-assay and inter-assay coefficient of variation were lower than 2.0%. Clinical specimens verified that the triplex qRT-PCR had a higher sensitivity and specificity than universal PCR. In conclusion, this triplex probe qRT-PCR could detect only BPV, BCoV and BPIV. Minimum detection limits were 2.0×102 copies/µL for BPV and BCoV, and 2.0×101 copies/µL for BPIV. The sensitivity of this triplex probe qRT-PCR was 1000-fold greater than that in the conventional PCR. The newly qRT-PCR could be used to monitor or differentially diagnose virus infection.


Assuntos
Bocavirus/isolamento & purificação , Coronavirus Bovino/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/veterinária , Vírus da Parainfluenza 3 Bovina/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Bocavirus/genética , Bovinos , Coronavirus Bovino/genética , DNA Viral/isolamento & purificação , Vírus da Parainfluenza 3 Bovina/genética , Plasmídeos/genética , RNA Viral/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
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