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使用16S rRNA基因测序和定量实时PCR分析患有牛蹄腐病和牛趾间皮炎病变的育肥牛的细菌群落。

Profiling bacterial communities in feedlot cattle affected with bovine foot rot and bovine digital dermatitis lesions using 16S rRNA gene sequencing and quantitative real-time PCR.

作者信息

Pyakurel Susan, Caddey Benjamin Jordan, Dias Angelica Petersen, De Buck Jeroen, Morck Douglas Walter, Orsel Karin

机构信息

Faculty of Veterinary Medicine, Health Sciences Center, University of Calgary, 3330 Hospital Drive, Calgary, NW, AB, T2N 4N1, Canada.

Department of Biological Sciences, Faculty of Science, University of Calgary, Calgary, Canada.

出版信息

BMC Microbiol. 2025 Mar 20;25(1):158. doi: 10.1186/s12866-025-03869-w.

Abstract

BACKGROUND

The primary infectious foot diseases in cattle, bovine foot rot (BFR) and bovine digital dermatitis (BDD), commonly associated with Fusobacterium necrophorum and Treponema spp., respectively, are considered polybacterial in etiology with several additional bacteria involved such as Porphyromonas levii, Bacteroides pyogenes, and Fusobacterium mortiferum. BDD is further classified into several M-stages (M2: active and ulcerative; M4: chronic proliferative). Using quantitative real-time PCR and 16S rRNA gene (V3-V4 region) sequencing, we quantified several specific bacteria and analyzed bacterial communities present in biopsies of visually diagnosed cases of BFR (n = 32), M2 (n = 17), and M4 (n = 12) stages of BDD in feedlot cattle in contrast to inconclusive (n = 14) clinical cases and healthy (n = 25) cattle.

RESULTS

Bacterial composition of healthy skin differed significantly from that of skin lesions, and between BFR and both lesion stages of BDD, which also differed from each other. All animal groups had generally the same bacterial species, albeit in distinct ratios. Differential abundance analysis relative to the healthy group identified a higher abundance of Fusobacterium spp. in BFR and Treponema spp. in both BDD-M2 and BDD-M4. P. levii had the highest absolute abundance in all animal groups. A significantly higher abundance of F. necrophorum was observed in BFR compared to BDD-M2, and F. mortiferum in both stages of BDD compared to the inconclusive group. Both BDD M-stages had a significantly higher abundance of Treponema phagedenis and Treponema pedis. Treponema medium was significantly more abundant in BDD-M4 compared to BDD-M2.

CONCLUSION

These results further the evidence of the involvement of Treponema spp., in BDD in feedlot cattle. However, it suggests further exploration of the role of Fusobacterium spp. in BFR and BDD. Importantly, a discriminating polybacterial involvement in these infections was evident demonstrated by changes in the population of multiple bacteria when compared to healthy animals.

摘要

背景

牛的主要传染性足部疾病,牛蹄腐病(BFR)和牛趾间皮炎(BDD),通常分别与坏死梭杆菌和密螺旋体属有关,病因被认为是多细菌的,还涉及其他几种细菌,如莱氏卟啉单胞菌、化脓拟杆菌和致死梭杆菌。BDD进一步分为几个M期(M2:活跃性和溃疡性;M4:慢性增生性)。我们使用定量实时PCR和16S rRNA基因(V3-V4区域)测序,对来自饲养场牛的经视觉诊断的BFR病例(n = 32)、BDD的M2期(n = 17)和M4期(n = 12)活检样本中的几种特定细菌进行了定量,并分析了细菌群落,同时与不确定的临床病例(n = 14)和健康牛(n = 25)进行了对比。

结果

健康皮肤的细菌组成与皮肤病变的细菌组成有显著差异,BFR与BDD的两个病变阶段之间也存在差异,且这两个病变阶段彼此也不同。所有动物组的细菌种类总体相同,尽管比例不同。相对于健康组的差异丰度分析表明,BFR中梭杆菌属的丰度较高,BDD-M2和BDD-M4中密螺旋体属的丰度较高。莱氏卟啉单胞菌在所有动物组中的绝对丰度最高。与BDD-M2相比,BFR中坏死梭杆菌的丰度显著更高,与不确定组相比,BDD两个阶段中致死梭杆菌的丰度更高。BDD的两个M期噬齿密螺旋体和足密螺旋体的丰度均显著更高。与BDD-M2相比,BDD-M4中中间密螺旋体的丰度显著更高。

结论

这些结果进一步证明了密螺旋体属在饲养场牛BDD中的参与。然而,这表明需要进一步探索梭杆菌属在BFR和BDD中的作用。重要的是,与健康动物相比,多种细菌数量的变化明显表明这些感染存在多细菌的区别性参与。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bbfd/11924851/83e6c34a2799/12866_2025_3869_Fig6_HTML.jpg

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