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肉牛弯曲杆菌的慢性排菌

Chronic shedding of Campylobacter species in beef cattle.

作者信息

Inglis G D, Kalischuk L D, Busz H W

机构信息

Agriculture and Agri-Food Canada Research Centre, Lethbridge, Canada.

出版信息

J Appl Microbiol. 2004;97(2):410-20. doi: 10.1111/j.1365-2672.2004.02313.x.

Abstract

AIMS

To determine the prevalence of chronic shedding of Campylobacter species by beef cattle, a longitudinal study of shedding patterns was conducted in a cohort of 60 beef steers over a 4-month period.

METHODS AND RESULTS

Steers were maintained in a simulated feedlot setting but individually in pens to minimize transmission among animals. At each collection time, campylobacters in faeces were detected using conventional PCR. In addition, quantities of Campylobacter jejuni and C. lanienae in faeces were measured using real-time quantitative (RTQ) PCR. All of the steers tested shed Campylobacter species during the course of the study, and overall, 90% of the 299 samples tested were positive for Campylobacter DNA. The majority of the animals (86%) shed campylobacters at >/=4 sample times. The most prevalent taxon detected in bovine faeces was C. lanienae (56% of samples) followed by C. jejuni (13%), C. hyointestinalis (8%), and C. fetus (2%). No C. coli was detected, and 13% of the faecal samples contained two or more of the above species. Seven (12%) and 34 (57%) animals shed C. jejuni and C. lanienae at >/=3 sample times, respectively. For both C. lanienae and C. jejuni, a substantial number of cells were detected in faeces using RTQ-PCR; 27% of the samples positive for C. jejuni contained populations >10(4) cells g(-1) (maximum of 5 x 10(5) cells g(-1)), and 44% of samples positive for C. lanienae possessed populations >10(6) cells g(-1) (maximum of 4 x 10(8) cells g(-1)). A significant correlation was observed between shedding of C. lanienae and the severity of liver abscesses. In 27% of the samples, an amplicon was obtained for genus-specific but not for the species-specific primers. Sequencing of the partial 16S rRNA gene suggested the presence of at least two undescribed Campylobacter species but this has yet to be confirmed.

CONCLUSIONS

A high percentage of feedlot cattle shed large quantities of Campylobacter species in their faeces over a protracted period of time (ca 112 days).

SIGNIFICANCE AND IMPACT OF THE STUDY

This is the first study of longitudinal shedding patterns of campylobacters in beef cattle using PCR-detection methods. In addition, this is the first use of RTQ-PCR to directly quantify C. jejuni or C. lanienae in faeces. The results of the study show that a large number of cattle (>85%) chronically shed campylobacters in feedlots.

摘要

目的

为确定肉牛弯曲杆菌慢性排菌的流行情况,对60头肉用公牛进行了为期4个月的排菌模式纵向研究。

方法与结果

公牛饲养在模拟饲养场环境中,但单独圈养以尽量减少动物间传播。每次采集粪便样本时,采用常规PCR检测弯曲杆菌。此外,采用实时定量(RTQ)PCR测定粪便中空肠弯曲杆菌和拉尼尔弯曲杆菌的数量。在研究过程中,所有检测的公牛均排出弯曲杆菌,总体而言,在检测的299份样本中,90%的样本弯曲杆菌DNA呈阳性。大多数动物(86%)在≥4个采样时间点排出弯曲杆菌。在牛粪中检测到的最常见分类群是拉尼尔弯曲杆菌(占样本的56%),其次是空肠弯曲杆菌(13%)、猪肠弯曲杆菌(8%)和胎儿弯曲杆菌(2%)。未检测到大肠弯曲杆菌,13%的粪便样本含有两种或更多上述菌种。分别有7头(12%)和34头(57%)动物在≥3个采样时间点排出空肠弯曲杆菌和拉尼尔弯曲杆菌。对于拉尼尔弯曲杆菌和空肠弯曲杆菌,使用RTQ-PCR在粪便中检测到大量细菌;空肠弯曲杆菌阳性样本中27%的菌量>10⁴ cfu/g(最大5×10⁵ cfu/g),拉尼尔弯曲杆菌阳性样本中44%的菌量>10⁶ cfu/g(最大4×10⁸ cfu/g)。观察到拉尼尔弯曲杆菌的排出与肝脓肿的严重程度之间存在显著相关性。在27%的样本中,获得了属特异性引物的扩增子,但未获得种特异性引物的扩增子。部分16S rRNA基因测序表明存在至少两种未描述的弯曲杆菌菌种,但尚未得到证实。

结论

在较长时间(约112天)内,很大比例的饲养场肉牛粪便中排出大量弯曲杆菌。

研究的意义和影响

这是首次采用PCR检测方法对肉牛弯曲杆菌纵向排菌模式进行的研究。此外,这是首次使用RTQ-PCR直接定量粪便中的空肠弯曲杆菌或拉尼尔弯曲杆菌。研究结果表明,大量肉牛(>85%)在饲养场中长期排出弯曲杆菌。

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