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拭子类型有影响吗?比较育肥牛中溶血曼氏杆菌的回收方法及上呼吸道微生物组特征

Does swab type matter? Comparing methods for Mannheimia haemolytica recovery and upper respiratory microbiome characterization in feedlot cattle.

作者信息

Crosby William B, Pinnell Lee J, Richeson John T, Wolfe Cory, Castle Jake, Loy John Dustin, Gow Sheryl P, Seo Keun Seok, Capik Sarah F, Woolums Amelia R, Morley Paul S

机构信息

Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA.

Veterinary Education, Research, and Outreach Program, Texas A&M University, VERO Building, 3201 Russell Long Blvd., Canyon, TX, 79015, USA.

出版信息

Anim Microbiome. 2022 Aug 13;4(1):49. doi: 10.1186/s42523-022-00197-6.

Abstract

BACKGROUND

Bovine respiratory disease (BRD) is caused by interactions among host, environment, and pathogens. One standard method for antemortem pathogen identification in cattle with BRD is deep-guarded nasopharyngeal swabbing, which is challenging, costly, and waste generating. The objective was to compare the ability to recover Mannheimia haemolytica and compare microbial community structure using 29.5 inch (74.9 cm) deep-guarded nasopharyngeal swabs, 16 inch (40.6 cm) unguarded proctology swabs, or 6 inch (15.2 cm) unguarded nasal swabs when characterized using culture, real time-qPCR, and 16S rRNA gene sequencing. Samples for aerobic culture, qPCR, and 16S rRNA gene sequencing were collected from the upper respiratory tract of cattle 2 weeks after feedlot arrival.

RESULTS

There was high concordance of culture and qPCR results for all swab types (results for 77% and 81% of sampled animals completely across all 3 swab types for culture and qPCR respectively). Microbial communities were highly similar among samples collected with different swab types, and differences identified relative to treatment for BRD were also similar. Positive qPCR results for M. haemolytica were highly concordant (81% agreed completely), but samples collected by deep-guarded swabbing had lower amounts of Mh DNA identified (Kruskal-Wallis analysis of variance on ranks, P < 0.05; Dunn-test for pairwise comparison with Benjamini-Hochberg correction, P < 0.05) and lower frequency of positive compared to nasal and proctology swabs (McNemar's Chi-square test, P < 0.05).

CONCLUSIONS

Though differences existed among different types of swabs collected from individual cattle, nasal swabs and proctology swabs offer comparable results to deep-guarded nasopharyngeal swabs when identifying and characterizing M. haemolytica by culture, 16S rRNA gene sequencing, and qPCR.

摘要

背景

牛呼吸道疾病(BRD)是由宿主、环境和病原体之间的相互作用引起的。在患有BRD的牛中,一种用于生前病原体鉴定的标准方法是深部鼻咽拭子采样,这种方法具有挑战性、成本高且会产生废弃物。目的是比较使用29.5英寸(74.9厘米)深部鼻咽拭子、16英寸(40.6厘米)无防护直肠拭子或6英寸(15.2厘米)无防护鼻拭子在通过培养、实时定量PCR和16S rRNA基因测序进行特征分析时,回收溶血曼氏杆菌的能力以及比较微生物群落结构。在牛进入饲养场2周后,从其上呼吸道采集用于需氧培养、定量PCR和16S rRNA基因测序的样本。

结果

所有拭子类型的培养和定量PCR结果具有高度一致性(分别在所有3种拭子类型中,77%和81%的采样动物结果完全一致)。不同拭子类型采集的样本中微生物群落高度相似,相对于BRD治疗所确定的差异也相似。溶血曼氏杆菌的定量PCR阳性结果高度一致(81%完全一致),但深部拭子采样采集的样本中鉴定出的溶血曼氏杆菌DNA量较低(Kruskal-Wallis秩和方差分析,P<0.05;与Benjamini-Hochberg校正的成对比较的Dunn检验,P<0.05),与鼻拭子和直肠拭子相比,阳性频率较低(McNemar卡方检验,P<0.05)。

结论

尽管从个体牛采集的不同类型拭子之间存在差异,但在通过培养、16S rRNA基因测序和定量PCR鉴定和表征溶血曼氏杆菌时,鼻拭子和直肠拭子与深部鼻咽拭子的结果相当。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/64fb/9375289/c2dbc36b1887/42523_2022_197_Fig1_HTML.jpg

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