An investigation into the microflora of heroin
An investigation into the microflora of heroin
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DOI：
10.1099/0022-1317-51-11-1001
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发表时间：
2002-11-01
影响因子：
3
通讯作者：
Ashton, JR
中科院分区：
文献类型：
--
作者：
McLauchlin, J;Mithani, V;Ashton, JR
2000年，据报道，英国和爱尔兰非法注射吸毒者的发病率和死亡率异常增加，诺维梭菌被确定为严重感染的可能来源，尽管也有肉毒杆菌和蜡样芽孢杆菌引起的感染的报道。由于海洛因是一种可能的感染源，本研究调查了 2000 年至 2002 年在英国缉获的海洛因样本的微生物群落。开发了两种检查海洛因微生物群落的方法。第一种方法是将药物悬浮在最大回收稀释剂 (MRD) 中，将其直接接种到肉毒杆菌分离熟肉汤 (CBI) 中。第二种方法使海洛因溶于柠檬酸，通过过滤浓缩颗粒材料（和细菌细胞），并通过用柠檬酸和磷酸盐缓冲盐水洗涤去除海洛因残留物，然后将过滤器放入CBI肉汤中。将两种方法的 CRI 肉汤副本在不加热的情况下以及在 60°C 加热 30 分钟后进行培养。孵育7天和14天后在八种不同的固体培养基上进行继代培养。使用掺有肉毒梭菌或诺维梭菌孢子悬浮液的海洛因样品对这些方法进行了评估；证明了原始样品中 10 个孢子的回收率。通过柠檬酸溶解技术测试了 58 个海洛因样品，通过 MRD 悬浮技术测试了 34 个海洛因样品。识别出四个属的十五种不同的革兰氏阳性菌。没有分离到真菌。需氧内生孢子形成细菌（芽孢杆菌属和浸软类芽孢杆菌）是分离的主要微生物群，并且从每个样品中分离出至少一种物种。 R cereus 是最常见的物种，从 95% 的样品中分离出来，其中 40% 的样品中分离出地衣芽孢杆菌。一到五个样品产生了凝结芽孢杆菌、侧孢芽孢杆菌、短小芽孢杆菌、枯草芽孢杆菌和浸软芽孢杆菌的培养物。葡萄球菌属从 23 个（40%）样本中分离出来； S. warneri 和 S. epidermidis 是最常见的，分别从 13 个（22%）和 6 个（10%）样本中培养出来。一两个样品产生了金黄色葡萄球菌、头状葡萄球菌和溶血葡萄球菌的培养物。检测到的其余菌群包括两个被产气荚膜梭菌污染的样品和两个被索氏梭菌或铽梭菌污染的样品。从 43 个样品 (74%) 中分离出多种细菌，从其余 15 个样品中分离出单一细菌。在 13 个样品中仅分离出蜡状芽孢杆菌，在一个样品中仅分离出枯草芽孢杆菌，在一个样品中仅分离出短小芽孢杆菌。没有从任何海洛因样品中分离出肉毒梭菌和诺维梭菌。给出了对海洛因微生物群落进行最佳检查的建议。
In 2000, an unusual increase of morbidity and mortality among illegal injecting drug users in the UK and Ireland was reported and Clostridium novyi was identified as the likely source of the serious infection, although infections due to C botulinum and Bacillus cereus were also reported. Because heroin was a possible source of infection, this study investigated the microflora of heroin samples seized in England during 2000 and 2002. Two methods were developed for the examination of the microflora of heroin. The first consisted of suspension of the drug in maximum recovery diluent (MRD) which was inoculated directly into Clostridium Botulinum Isolation Cooked Meat Broth (CBI). The second method rendered the heroin soluble in citric acid, concentrated particulate material (and bacterial cells) by filtration and removed heroin residues by washing with citric acid and phosphate-buffered saline before placing the filter in CBI broth. Duplicate CRI broths from both methods were incubated without heating and after heating at 60degreesC for 30 min. Subcultures were made after incubation for 7 and 14 days on to eight different solid media. The methods were evaluated with heroin samples spiked with either C. botulinum or C. novyi spore suspensions; recovery of 10 spores in the original sample was demonstrated. Fifty-eight heroin samples were tested by citric acid solubilisation and 34 by the MRD suspension technique. Fifteen different gram-positive species of four genera were recognised. No fungi were isolated. Aerobic endospore-forming bacteria (Bacillus spp. and Paenibacillus macerans) were the predominant microflora isolated and at least one species was isolated from each sample. R cereus was the most common species and was isolated from 95% of all samples, with B. licheniformis isolated from 40%. Between one and five samples yielded cultures of B. coagulans, B. laterosporus, B. pumilus, B. subtilis and R macerans. Staphylococcus spp. were isolated from 23 (40%) samples; S. warneri and S. epidermidis were the most common and were cultured from 13 (22%) and 6 (10%) samples respectively. One or two samples yielded cultures of S. aureus, S. capitis and S. haemolyticus. The remainder of the flora detected comprised two samples contaminated with C perfringens and two samples with either C sordellii or C tertium. Multiple bacterial species were isolated from 43 (74%) samples, a single species from the remaining 15. In 13 samples B. cereus alone was isolated, in one B. subtilis alone and in one sample B. pumilus alone. C botulinum and C novyi were not isolated from any of the heroin samples. Recommendations for the optimal examination of the microflora of heroin are given.