RESUMO
Background/aim/AIM: SARS-CoV-2 disease was announced as a pandemic by The World Health Organization in early 2020. It is still threatening the world population. Here, we aimed to produce hyperimmune sera that contain immunoglobulin G and F(ab')2 fragments sourced from horse antibodies as an urgent response to the pandemic. Materials and methods: SARS-CoV-2 was produced and inactivated with three different methods [formaldehyde (FA), formaldehyde, and binary ethylene amine (FA + BEI), and heat treatment]. After in vitro inactivation control, immunogens were mixed with Freund's adjuvant, thereafter horses (n: 2 for FA, 4 for FA + BEI, 2 for heat inactivation) and New Zealand rabbits (n: 6 for FA, 6 fo r FA + BEI, 6 for heat inactivation) were immunized four times. Neutralizing antibody levels of the sera were measured at the 4th, 6th, and 8th weeks. When the antibodies were detected at the peak level, plasma was collected from horses and hyperimmune sera procured after the purification process. Results: Horses and rabbits produced highly neutralizing antibodies against the SARS-CoV-2 in FA and FA + BEI inactivation groups, foreign proteins were removed effectively after purification. Conclusion: This study presents a profitable practice to develop specific antisera in horses against SARS-CoV-2 for emergency and low-cost response. In further studies, new purification methods can be used to increase the efficiency of the final product.
Assuntos
Soros Imunes/farmacologia , Fatores Imunológicos/farmacologia , SARS-CoV-2/efeitos dos fármacos , Animais , Cavalos , Coelhos , Tratamento Farmacológico da COVID-19RESUMO
Bluetongue is a significant arbovirus infection that has a negative impact on ruminant productivity in Turkey. Twenty-one Turkish BTV isolates were analyzed phylogenetically, based on genome segment 10 (Seg-10) nucleotide sequences. These analyses were used to explore the epidemiological background of individual isolates from both a regional and global perspective. In the regional analysis, the different BTV strains fell into two groups (Group 1 and Group 2). The Turkish virus isolates were localized in Group 1 which contains two sub-groups. The neighbor-joining analysis revealed that Seg-10 of majority of the Turkish viruses was closely related to certain other virus strains allocated in the eastern lineage. The Seg-10's of two viruses (TR25 and TR26) were more closely related to strains isolated in the Asia-Australia region. These strains belong to the 'eastern' topotype identified by [Maan, S., Maan, N.S., Ross-Smith, N., Batten, C.A., Shaw, A.E., Anthony, S.J., Samuel, A.R., Darpel, K.E., Veronesi, E., Oura, C.A.L., Singh,K.P., Nomikou, K., Potgieter, A.C., Attoui, H., van Rooij, E., van Rijn, P., De Clercq, K., Vandenbussche, F., Zientara, S., Bréard, E., Sailleau, C., Beer, M., Hoffman, B., Mellor, P.S., Mertens, P.P.C., 2008. Sequence analysis of bluetongue virus serotype 8 from the Netherlands 2006 and comparison to other European strains. Virology 377, 308-318]. Comparisons of amino acid sequences deduced from the Seg-10 genes showed a high level of conservation in the NS3/3A proteins from the Turkish viruses. The more frequent amino acid substitutions were identified by multiple alignment analysis, and one of the isolates (TR23) was remarkably found to be genetically quite distinct from the other isolates.
Assuntos
Vírus Bluetongue/genética , Vírus Bluetongue/isolamento & purificação , Bluetongue/epidemiologia , Sequência de Aminoácidos , Animais , Bluetongue/virologia , Vírus Bluetongue/química , Vírus Bluetongue/classificação , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Alinhamento de Sequência , Turquia/epidemiologia , Proteínas Virais/química , Proteínas Virais/genéticaRESUMO
Group A rotaviruses are major enteric pathogens of calves. In order to investigate the genetic diversity of bovine rotaviruses (BRVs), a collection of 53 BRVs, detected from diarrheic calves from several Turkish geographical areas, between 1997 and 2008 was analyzed by RT-PCR for specificities of the outer capsid proteins VP7 (G type) and VP4 (P type), for the first time. Overall, G6 was the predominant G type, detected in 40/53 samples (75.4%), while P[11] was the predominant P type, detected in 52/53 samples (98.1%). The most common VP7/VP4 combinations were G6P[11] (60.3%) and G10P[11] (24.5%). Mixed infections were identified in 7/53 samples (13.2%). In the VP7 region the G6P[11] viruses were similar to other ones detected worldwide, forming an independent G6 lineage, distantly related to the G6 gene of the vaccine G6P[1] strain NCDV (90.1% amino acid identity), and suggesting that G6P[11] viruses represent a genetically stable BRV strain. The study of G and P type diversity is pivotal to understand the efficacy of the existing rotavirus vaccines and to provide the basis of future prophylaxis tools against rotaviral diarrhea of calves.