Seroprevalence and associated factors of viral agents of the bovine respiratory disease complex in buffaloes of Colombia

ABSTRACT: The bovine respiratory disease complex causes sizable losses for livestock producers. Its presentation involves the interaction among multiple viral and bacterial pathogens. Viral agents include: bovine viral diarrhea virus (BVDV), bovine alphaherpesvírus 1 (BoHV-1), bovine parainfluenza virus (bPI3v) and bovine respiratory syncytial virus (BRSV); so far, these viruses have been little studied in the buffalo population. This study is aimed to assess the presence of these viruses in buffalo populations of the department of Córdoba, Colombia. A transversal study was conducted upon assessment of 37 farms and 861 buffaloes. Seroprevalence in animals were as follows: 21.7% (BVDV), 51.5% (BoHV-1), 73.6% (bPI3v) and 58.9% (BSRV), while seroprevalence in farms stood at: 94.6% (BVDV), 100% (BoHV-1), 100% (bPI3v) and 100% (BSRV). Conversely, the multiple seropositivity in buffaloes or past coinfection of viral agents amounted to 23.3% for a single virus, 31.8% for two viruses, 29.4% for three viruses and 7.1% for four viruses; only 8.4% of buffaloes were seronegative for four viruses. This is the first regional epidemiological study in the buffalo populations of Colombia and the results showed viral circulation and multiple seropositivity in animals; in addition, the high prevalence found pose a concerning epidemiological threat.

Quantitative Proteomic Analysis Shows Involvement of the p38 MAPK Pathway in Bovine Parainfluenza Virus type 3 Replication

Background: Bovine parainfluenza virus type 3 (BPIV3) infection often causes respiratory tissue damage and immunosuppression and results in bovine respiratory disease complex. Bovine respiratory disease complex is one of the major diseases in dairy cattle and causes huge economical losses every year. The pathogenetic and immunoregulatory mechanisms involved in the process of BPIV3 infection, however, remain unknown. Proteomics is a powerful tool for high-throughput identification of proteins and 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 MAPKpathway to promote virus replication.

Bovine Parainfluenza Virus Type 3 (BPIV3) Enters HeLa Cells via Clathrin-Mediated Endocytosis in a Cholesterol- and Dynamin-Dependent Manner

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 MβCD 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.

Quantitative Proteomic Analysis Shows Involvement of the p38 MAPK Pathway in Bovine Parainfluenza Virus Type 3 Replication

Bovine parainfluenza virus type 3 (BPIV3) infection often causes respiratory tissue damage and immunosuppression and results in bovine respiratory disease complex. Bovine respiratory disease complex is one of the major diseases in dairy cattle and causes huge economical losses every year. The pathogenetic and immunoregulatory mechanisms involved in the process of BPIV3 infection, however, remain unknown. Proteomics is a powerful tool for high-throughput identification of proteins and has been widely used to understand how viruses interact with host cells. 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 invasion 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. 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. The results of proteomics analysis indicated BPIV3 infection could activate the p38 MAPK pathway to promote virus replication.

Evolution of the Seroprevalence of Pestivirus and Respiratory Viral Infections in Spanish Feedlot Lambs

The presence of respiratory viruses and pestiviruses in sheep has been widely demonstrated, and their ability to cause injury and predispose to respiratory processes have been proven experimentally. A longitudinal observational study was performed to determine the seroprevalence of bovine parainfluenza virus type 3 (BPIV-3), bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1 (BHV-1) and pestiviruses in 120 lambs at the beginning and the end of the fattening period. During this time, the animals were clinically monitored, their growth was recorded, and post-mortem examinations were performed in order to identify the presence of pneumonic lesions in the animals. Seroconversion to all viruses tested except BHV-1 was detected at the end of the period. Initially, BPIV-3 antibodies were the most frequently found, while the most common seroconversion through the analysed period occurred to BRSV. Only 10.8% of the lambs showed no detectable levels of antibodies against any of the tested viruses at the end of the survey. In addition, no statistical differences were found in the presentation of respiratory clinical signs, pneumonic lesions nor in the production performance between lambs that seroconverted and those which did not, except in the case of pestiviruses. The seroconversion to pestiviruses was associated with a reduction in the final weight of the lambs.