Department of Clinical Veterinary Studies, Faculty of Veterinary Science, University of Zimbabwe, Harare, Zimbabwe.
J Appl Microbiol. 2012 Dec;113(6):1389-95. doi: 10.1111/jam.12006. Epub 2012 Oct 1.
To isolate Bacillus anthracis from cattle carcass burial sites from high-risk districts in Zimbabwe.
Soil samples were collected from carcass burial sites from seven areas, including two national game parks. Samples were collected from top 5-10 cm, and for spore extraction, 25 g of soil was suspended in sterile distilled water overnight. Supernatants were filtered through 0.45-μm pore cellulose nitrate, deposits suspended in 5 ml phosphate-buffered saline, aliquoted and heated at temperature regimen of 65, 70, 75 and 80 °C for 15 min. Samples were plated onto PLET agar. B. anthracis isolates were identified using growth morphology and PCR detecting pXO1 and pXO2 virulence plasmids. From samples heated at 75 °C for 15 min, B. anthracis were isolated from 9 of 81 (11.1%) soil samples representing five of the seven sampled areas.
We isolated B. anthracis from soil collected from carcass burial sites. PCR targeting virulence plasmids provided a rapid confirmation of B. anthracis.
The positive isolation indicated that some carcass burial sites may retain viable spores for at least 12 months after the previous outbreak, which suggests that they may be important sources of B. anthracis and new disease outbreaks.
从津巴布韦高风险地区的牛尸埋葬地点分离炭疽杆菌。
从七个地区(包括两个国家野生动物园)的尸体埋葬地点采集土壤样本。采集表层 5-10cm 的土壤,为提取孢子,将 25g 土壤悬浮于无菌蒸馏水,过夜。上清液通过 0.45-μm 孔径的硝酸纤维素过滤,沉淀物悬浮于 5ml 磷酸盐缓冲盐水,等分并加热至 65、70、75 和 80°C 15 分钟。将样品接种于 PLET 琼脂。使用生长形态和 PCR 检测 pXO1 和 pXO2 毒力质粒鉴定炭疽杆菌分离株。从加热至 75°C 15 分钟的样品中,从代表七个采样区中的五个采样区的 81 个(11.1%)土壤样品中的 9 个分离出炭疽杆菌。
我们从尸体埋葬地点采集的土壤中分离出炭疽杆菌。针对毒力质粒的 PCR 快速确认了炭疽杆菌。
阳性分离表明,在前一次疫情发生后至少 12 个月,一些尸体埋葬地点可能仍保留有存活孢子,这表明它们可能是炭疽杆菌和新疾病爆发的重要来源。