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1.
Int J Mol Sci ; 25(14)2024 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-39062990

RESUMO

Johne's disease (JD), also known as paratuberculosis, is a chronic, untreatable gastroenteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP) infection. Evidence for host genetic resistance to disease progression exists, although it is limited due to the extended incubation period (years) and diagnostic challenges. To overcome this, previously restored formalin-fixed paraffin embedded tissue (FFPE) DNA from archived FFPE tissue cassettes was utilized for a novel retrospective case-control genome-wide association study (GWAS) on ovine JD. Samples from known MAP-infected flocks with ante- and postmortem diagnostic data were used. Cases (N = 9) had evidence of tissue infection, compared to controls (N = 25) without evidence of tissue infection despite positive antemortem diagnostics. A genome-wide efficient mixed model analysis (GEMMA) to conduct a GWAS using restored FFPE DNA SNP results from the Illumina Ovine SNP50 Bead Chip, identified 10 SNPs reaching genome-wide significance of p < 1 × 10-6 on chromosomes 1, 3, 4, 24, and 26. Pathway analysis using PANTHER and the Kyoto Encyclopedia of Genes and Genomes (KEGG) was completed on 45 genes found within 1 Mb of significant SNPs. Our work provides a framework for the novel use of archived FFPE tissues for animal genetic studies in complex diseases and further evidence for a genetic association in JD.


Assuntos
Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Inclusão em Parafina , Paratuberculose , Polimorfismo de Nucleotídeo Único , Doenças dos Ovinos , Animais , Paratuberculose/genética , Paratuberculose/microbiologia , Ovinos , Doenças dos Ovinos/genética , Doenças dos Ovinos/microbiologia , Estudos Retrospectivos , Mycobacterium avium subsp. paratuberculosis/genética , DNA/genética , Formaldeído , Estudos de Casos e Controles , Resistência à Doença/genética
2.
Vet Res Commun ; 47(1): 131-139, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35618986

RESUMO

Archived formalin fixed paraffin-embedded (FFPE) tissues are powerful tools in medicine, capable of harboring diagnostic and genetic answers to challenging clinical questions. Successful utilization of DNA derived from FFPE samples is dependent upon repairing DNA damage generated from the fixation process. Methods to repair FFPE DNA have been successful in human medicine for a variety of research and clinical applications, yet remain underutilized in veterinary medicine. Despite the available technology, our study is the first to evaluate the repair of FFPE derived DNA from veterinary species for single-nucleotide polymorphism (SNP) analysis using the Illumina OvineSNP50 BeadChip and Illumina FFPE QC and DNA Restore kit. To accomplish this, 48 ovine FFPE samples were run using the Illumina OvineSNP50 BeadChip with and without restoration. Compared to pre-restore data, we found increased sample call rates, SNP call frequency, and assay metrics for all samples post-restoration. Further, we utilized four sheep with available parallel fresh DNA and FFPE DNA to compare assay metrics and genotype calls between the two starting sample types. Although fresh samples generated increased call rates, we found 99% concordance in allele calls between restored FFPE and fresh DNA for all four samples. Our results indicate successful restoration and genotyping of ovine FFPE samples using this technology, with potential for utilization in other veterinary species.


Assuntos
Formaldeído , Polimorfismo de Nucleotídeo Único , Humanos , Animais , Ovinos/genética , Fixação de Tecidos/veterinária , Inclusão em Parafina/veterinária , DNA/genética
3.
Trop Anim Health Prod ; 50(4): 793-799, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29302775

RESUMO

A cross-sectional study was carried out to identify risk factors for bovine viral diarrhea virus (BVDV) infection in 62 randomly selected dairy herds which were tested for BVD serum antibodies by using an indirect ELISA kit (IDEXX). Results from the chi-square test analysis were interpreted by analyzing by chi-square test. A sum of 500 sera samples were screened and 66 animals (13.20%) showed positive for BVDV antibody. Within herd, BVD seroprevalence was 12-65%. This study concluded that epidemiological risk factors like location, herd size, housing patterns like, tail to tail system, roofing pattern, distance between the manure pit and farm, and distance between farms were significantly associated with BVDV serological status (P < 0.05).


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Animais , Anticorpos Antivirais/sangue , Bovinos , Estudos Transversais , Diarreia , Vírus da Diarreia Viral Bovina Tipo 1 , Ensaio de Imunoadsorção Enzimática/veterinária , Fazendas , Feminino , Índia/epidemiologia , Fatores de Risco , Estudos Soroepidemiológicos
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