Kurzejamska-Parafiniuk M
Katedry i Zakładu Medycyny Sadowej Pomorskiej Akademii Medycznej w Szczecinie.
Ann Acad Med Stetin. 1996;42:85-104.
Evaluating the results of sectional blood examinations for ethyl alcohol content is difficult due to the proceeding putrid and fermentative processes, in consequence of which endogenic ethyl alcohol is produced. Some difficulties also arise from estimating the results of serological investigations concerning the biological traces having been changed by putrefaction, where the presence of heterogenic antigens may be suspected. The putrid-fermentative processes are linked with the activity of microorganisms, particularly bacteria and yeast-like fungi. The first part of the paper deals with the bacterial flora in 50 sectional blood samples taken for routine determinations of ethyl alcohol content, thus with added sodium fluoride as bacteriostatic agent. The identification of the microorganisms cultured on differentiating and selectively differentiating media was carried out on the basis of the culture appearance, specimens stained by Gram's method, as well as biochemical examinations. From 16 studied samples of the sectional blood no strain of bacteria was cultured, mixed bacterial flora was isolated from the remaining ones (Tab. 1). Most numerous were Gram-negative bacteria (71%) among which E.coli appeared most frequently. Gram-positive claimed 28% of the cultured microflora, while anaerobes hardly 4%. In the second part of the paper, the selected strains of E. coli pertaining to serological groups: 02, 04, 06, 08, 09, 022, 025 were studied with regard to their possibility to produce ethanol as well as antigens A and B of AB0 system. E.coli strains were grown on broth medium containing glucose in concentration of from 0.00 to 27.75 mmol/l and human blood of 0 group collected from blood-donors on sodium citrate and CPD preservative with glucose in its content. Ethanol concentration in cultures was determined after 24 and 72 hours of incubation, by gas chromatography method, and glucose by enzymatic method. In serological investigations the material consisted of linen pieces being covered with 72-hour cultures of E.coli on broth and human blood of group 0. The study for the presence of the group substances A and B of AB0 system was performed by absorption method according to Holzer, and by absorption and elution method. In consequence of the studies it has been ascertained that sodium fluoride added as a bacteriostatic agent does not entirely inhibit the growth of bacteria, and especially bacteria Gram-negative appeared to be least sensitive to its action (Tab. 1). Selected strains of E.coli have differed with regard to the efficiency of ethanol production (Tab. 2). The level of produced ethanol depended on glucose concentration in the medium, temperature and the incubation time (Tab. 2, 3, 4 and Fig. 1). Under almost similar conditions the same strains produce more alcohol than on blood, which may give rise to supposition that the blood modifies the metabolism of bacteria (Tab. 4 and 5). The cultures of selected strains studied failed to reveal the presence of heteroantigens with properties of antigen A and B of AB0 system.
由于存在腐败和发酵过程,会产生内源性乙醇,因此评估血液中乙醇含量的分段检测结果很困难。在估计因腐败而发生变化的生物痕迹的血清学调查结果时也会出现一些困难,在这些调查中可能会怀疑存在异源抗原。腐败发酵过程与微生物的活动有关,特别是细菌和酵母样真菌。本文的第一部分研究了50份用于常规乙醇含量测定的分段血液样本中的细菌菌群,这些样本添加了氟化钠作为抑菌剂。在鉴别和选择性鉴别培养基上培养的微生物的鉴定是根据培养物外观、革兰氏染色标本以及生化检查进行的。在16份研究的分段血液样本中未培养出细菌菌株,其余样本分离出混合细菌菌群(表1)。革兰氏阴性菌最多(71%),其中大肠杆菌出现频率最高。革兰氏阳性菌占培养菌群的28%,厌氧菌几乎占4%。在本文的第二部分,研究了属于血清学组02、04、06、08、09、022、025的大肠杆菌选定菌株产生乙醇以及AB0系统抗原A和B的可能性。大肠杆菌菌株在含有浓度为0.00至27.75 mmol/l葡萄糖的肉汤培养基以及从使用柠檬酸钠和CPD防腐剂且含有葡萄糖的献血者采集的0型人血中生长。培养24小时和72小时后,通过气相色谱法测定培养物中的乙醇浓度,通过酶法测定葡萄糖浓度。在血清学研究中,材料包括覆盖有大肠杆菌在肉汤和0型人血中72小时培养物的亚麻布片。根据霍尔泽的吸收法以及吸收和洗脱法对AB0系统中A和B组物质的存在进行研究。研究结果表明,添加作为抑菌剂的氟化钠并不能完全抑制细菌生长,尤其是革兰氏阴性菌似乎对其作用最不敏感(表1)。选定的大肠杆菌菌株在乙醇产生效率方面存在差异(表2)。产生的乙醇水平取决于培养基中的葡萄糖浓度、温度和培养时间(表2、3、4和图1)。在几乎相同的条件下,相同菌株在培养基上产生的酒精比在血液中更多,这可能会让人推测血液会改变细菌的代谢(表4和5)。对选定菌株的培养物进行研究未发现具有AB0系统抗原A和B特性的异源抗原。