Mazzei Maurizio, Sorvillo Benedetta, Sgorbini Micaela, Bindi Francesca, Perelli Alice, Laus Fulvio
Department of Veterinary Sciences, University of Pisa, Viale delle Piagge 2, 56124 Pisa, Italy.
School of Biosciences and Veterinary Medicine, University of Camerino, Via Pontoni 5, 62032 Camerino, Italy.
Animals (Basel). 2024 Nov 21;14(23):3347. doi: 10.3390/ani14233347.
(1) Background: Respiratory problems may be associated with pathogens among which viruses may play an important role in causing or promoting clinical signs. However, traditional diagnostic methods for equine infectious diseases, such as PCR, are limited to known pathogens and often miss rare or novel viruses. Metagenomic next-generation sequencing is a transformative method that allows the sequencing of all genomes present in a sample. Recent studies have used next-generation sequences to identify viral sequences in horses with unexplained respiratory symptoms, revealing potential links between viruses and respiratory problems. The aim of the study was to verify a relationship between respiratory diseases and the presence of viral agents through molecular analyses, applied to bronchoalveolar lavage fluid obtained from horses with history or presence of clinical signs of respiratory diseases. (2) Methods: A cohort of 14 horses was enrolled. All the subjects were submitted to a clinical assessment of the respiratory tract, thoracic ultrasound evaluation, respiratory tract endoscopy, and bronchoalveolar lavage. The latter one was used for cytological analysis, DNA and RNA extraction, and molecular biology analysis. (3) Results: No positive results were obtained in the molecular studies except for a sequence of 753 bp obtained by next-generation sequences, with complete homology to Equid gammaherpesvirus 2 strains. The samples were taken from a thoroughbred female horse aged 2 years old and referred for poor performance and sporadic cough at the beginning of the training session. (4) Conclusions: viral involvement may be not common in horses with respiratory diseases. We cannot exclude that the lack of virus detection may be due to the small sample size of the population included.
(1) 背景：呼吸问题可能与病原体有关，其中病毒在引发或加剧临床症状方面可能起重要作用。然而，马传染病的传统诊断方法，如聚合酶链反应（PCR），仅限于已知病原体，常常遗漏罕见或新型病毒。宏基因组下一代测序是一种变革性方法，可对样本中存在的所有基因组进行测序。最近的研究已利用下一代测序来鉴定有不明呼吸道症状马匹中的病毒序列，揭示了病毒与呼吸问题之间的潜在联系。本研究的目的是通过分子分析来验证呼吸道疾病与病毒病原体存在之间的关系，该分析应用于从有呼吸道疾病病史或临床症状的马匹获取的支气管肺泡灌洗液。(2) 方法：纳入了一组14匹马。所有受试者均接受了呼吸道临床评估、胸部超声检查、呼吸道内窥镜检查以及支气管肺泡灌洗。后者用于细胞学分析、DNA和RNA提取以及分子生物学分析。(3) 结果：分子研究未获得阳性结果，仅通过下一代测序获得了一段753 bp的序列，与马γ疱疹病毒2株完全同源。样本取自一匹2岁的纯种雌马，该马在训练初期因表现不佳和偶发咳嗽前来就诊。(4) 结论：病毒感染在患有呼吸道疾病 的马匹中可能并不常见。我们不能排除未检测到病毒可能是由于所纳入群体的样本量较小所致。
