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基于 16S rDNA 高通量测序鉴定犬外耳炎相关细菌。

Identification of bacteria associated with canine otitis externa based on 16S rDNA high-throughput sequencing.

机构信息

Princess Srisavangavadhana College of Medicine, Chulabhorn Royal Academy, Bangkok, 10210, Thailand.

Division of Biochemistry, Department of Preclinical Science, Faculty of Medicine, Thammasat University, Pathum Thani, 10120, Thailand.

出版信息

Braz J Microbiol. 2023 Dec;54(4):3283-3290. doi: 10.1007/s42770-023-01166-0. Epub 2023 Oct 27.

Abstract

Bacteria are regarded as predisposing and perpetuating factors causing otitis externa (OE), whereas auricular anatomy is a predisposing factor. This study aims to investigate bacterial populations in the external auditory canals of healthy dogs and dogs with OE. Four categories of ear swabs included healthy erect-ear dogs, erect-ear dogs with OE, healthy pendulous-ear dogs and pendulous-ear dogs with OE. After bacterial DNA extraction, 16S rDNAs were amplified using specific primers within a V3/V4 region. Following DNA library construction, high-throughput sequencing was performed on MiSeq (Illumina). CLC Microbial Genomics Module was used to determine the rarefaction curve, bacterial classification, relative abundance, richness and diversity index. The results demonstrated that healthy dogs had higher bacterial richness and diversity than the dogs with OE. Comparable with culture-dependent methods described previously, this study revealed predominant Corynebacterium spp., Pseudomonas spp., Staphylococcus spp., and Proteus spp. in OE cases. Furthermore, high-throughput sequencing might disclose some potential emerging pathogens including Tissierella spp., Acinetobacter spp., and Achromobacter spp., which have not been reported in previous canine OE cases. Nevertheless, larger sample sizes are further required for an extensive evidence-based investigation.

摘要

细菌被认为是引起外耳炎(OE)的诱发和持续因素,而耳廓解剖结构是诱发因素。本研究旨在调查健康犬和患有 OE 的犬的外耳道中的细菌种群。四类耳拭子包括健康直立耳犬、患有 OE 的直立耳犬、健康垂耳犬和患有 OE 的垂耳犬。在提取细菌 DNA 后,使用特定引物扩增 16S rDNA,该引物位于 V3/V4 区域内。在构建 DNA 文库后,在 MiSeq(Illumina)上进行高通量测序。CLC 微生物基因组学模块用于确定稀疏曲线、细菌分类、相对丰度、丰富度和多样性指数。结果表明,健康犬的细菌丰富度和多样性高于患有 OE 的犬。与以前描述的基于培养的方法类似,本研究在外耳炎病例中发现了主要的棒状杆菌属、假单胞菌属、葡萄球菌属和变形杆菌属。此外,高通量测序可能会揭示一些以前未在犬 OE 病例中报道的潜在新兴病原体,包括 Tissierella 属、不动杆菌属和无色杆菌属。然而,需要进一步扩大样本量以进行广泛的基于证据的调查。

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