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2018年至2022年下奥地利州通过基于DNA的检测(无需芽孢形成)进行的监测。

Monitoring of in Lower Austria through DNA-Based Detection without De-Sporulation: 2018 to 2022.

作者信息

Wilhelm Elfriede, Korschineck Irina, Sigmund Michael, Paulsen Peter, Hilbert Friederike, Rossmanith Wigbert

机构信息

NOE-Animal Health Service, Schillerring 13, 3130 Herzogenburg, Austria.

Ingenetix GmbH, Arsenalstraße 11, 1030 Vienna, Austria.

出版信息

Vet Sci. 2023 Mar 10;10(3):213. doi: 10.3390/vetsci10030213.

Abstract

American foulbrood is caused by the spore-forming . Although the disease effects honey bee larvae, it threatens the entire colony. Clinical signs of the disease are seen at a very late stage of the disease and bee colonies are often beyond saving. Therefore, through active monitoring based on screening, an infection can be detected early and bee colonies can be protected with hygiene measures. As a result, the pressure to spread in an area remains low. The cultural and molecular biological detection of is usually preceded by spore germination before detection. In this study, we compared the results of two methods, the culture detection and RT-PCR detection of DNA directly isolated from spores. Samples of honey and cells with honey surrounding the brood were used in a five-year voluntary monitoring program in a western part of Lower Austria. DNA-extraction from spores to speed up detection involved one chemical and two enzymatic steps before mechanical bashing-beat separation and additional lysis. The results are comparable to culture-based methods, but with a large time advantage. Within the voluntary monitoring program, the proportion of bee colonies without the detection of was high (2018: 91.9%, 2019: 72.09%, 2020: 74.6%, 2021: 81.35%, 2022: 84.5%), and in most -positive bee colonies, only a very low spore content was detected. Nevertheless, two bee colonies in one apiary with clinical signs of disease had to be eradicated.

摘要

美洲幼虫腐臭病由形成孢子的[病原体名称未给出]引起。虽然这种疾病影响蜜蜂幼虫,但它威胁着整个蜂群。该病的临床症状在疾病的非常晚期才会出现,蜂群往往无法挽救。因此,通过基于筛查的主动监测,可以早期检测到感染,并通过卫生措施保护蜂群。结果,该区域内的传播压力仍然较低。[病原体名称未给出]的培养和分子生物学检测通常在检测前先进行孢子萌发。在本研究中,我们比较了两种方法的结果,即从孢子直接分离的DNA的培养检测和RT-PCR检测。在奥地利下奥地利州西部的一项为期五年的自愿监测计划中,使用了蜂蜜样本以及育虫周围带有蜂蜜的巢脾样本。为了加快检测速度,从孢子中提取DNA在机械捣碎分离和额外裂解之前涉及一个化学步骤和两个酶促步骤。结果与基于培养的方法相当,但具有很大的时间优势。在自愿监测计划中,未检测到[病原体名称未给出]的蜂群比例很高(2018年:91.9%,2019年:72.09%,2020年:74.6%,2021年:81.35%,2022年:84.5%),并且在大多数呈阳性的蜂群中,仅检测到非常低的孢子含量。尽管如此,一个养蜂场中有两个出现疾病临床症状的蜂群不得不被根除。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/28cd/10054382/ee25af9168d4/vetsci-10-00213-g001.jpg

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