Lenchenko Ekaterina, Blumenkrants Dmitry, Sachivkina Nadezhda, Shadrova Nadezhda, Ibragimova Alfia
Department of Veterinary Medicine, Moscow State University of Food Production, Moscow, Russia.
Department of Microbiology and Virology, Medical Institute, Peoples' Friendship University of Russia, Moscow, Russia.
Vet World. 2020 Jan;13(1):197-200. doi: 10.14202/vetworld.2020.197-200. Epub 2020 Jan 28.
The study of biofilm-forming ability of Gram-negative microflora has great practical importance for assessing the effectiveness of antibiotic therapy and finding new ways to diagnose and inhibit the growth of biofilms. This is because poor penetration of antibacterial drugs into the biofilm can lead to the selection of resistant strains and has a consequence evident by the occurrence of relapse of infection in animals. This study aimed to evaluate morphological and densitometric indicators of biofilm formation as well as adhesive properties of .
was cultured at 37°C for 2-144 h . The specimens for optical microscopy were prepared by fixation with a 1:1 alcohol-ether mixture for 10 min and stained with a 0.5% solution of gentian violet for 2 min, and the optical density index was evaluated at a wavelength of 490 nm. Further, the adhesive properties of the microorganisms were determined at a concentration of 1 billion/ml and a suspension of ram erythrocytes at a concentration of 100 million/ml when cultured at 37°C for 24 h. Blood smears were prepared and stained with 0.5% gentian violet.
cultured at 37°C after 24 h on the meat peptone agar formed large, convex, mucous, and white colonies (d=3.0-6.0 mm). With the growth in the meat and peptone broth, uniform turbidity of the medium was observed. Analyzing the optical density indices (density, D), it was found that were good producers of biofilms (D=0.528±0.31). Data for indicators of adhesive properties of were as follows: Average adhesion index, 4.56±0.14; adhesion coefficient, 1.07±0.52; and adhesion index, 4.26±0.07. The studied bacteria had high adhesive activity. A direct correlation dependence (R=0.94) of the optical density of biofilms (D≥0.514-0.551) and AAI (4.15±0.28-4.76±0.75) was established.
This study has demonstrated that had high adhesive activity, was strong producer of biofilms, and the optical density of the sample exceeded the optical density of the control by more than 4 times.
革兰氏阴性微生物生物膜形成能力的研究对于评估抗生素治疗效果以及寻找诊断和抑制生物膜生长的新方法具有重要的实际意义。这是因为抗菌药物难以穿透生物膜,可能导致耐药菌株的产生,其后果表现为动物感染复发。本研究旨在评估生物膜形成的形态学和光密度指标以及[具体微生物名称未给出]的黏附特性。
[具体微生物名称未给出]在37°C下培养2 - 144小时。光学显微镜检查标本的制备方法为:用1:1的酒精 - 乙醚混合物固定10分钟,然后用0.5%的龙胆紫溶液染色2分钟,并在490nm波长下评估光密度指数。此外,在浓度为10亿/ml时测定微生物的黏附特性,同时在37°C下培养24小时时,使用浓度为1亿/ml的绵羊红细胞悬液。制备血涂片并用0.5%的龙胆紫染色。
[具体微生物名称未给出]在37°C下于肉蛋白胨琼脂上培养24小时后形成大的、凸起的、黏液状的白色菌落(直径d = 3.0 - 6.0mm)。在肉蛋白胨肉汤中生长时,观察到培养基均匀浑浊。分析光密度指数(密度,D)发现,[具体微生物名称未给出]是生物膜的良好生产者(D = 0.528±0.31)。[具体微生物名称未给出]黏附特性指标的数据如下:平均黏附指数为4.56±0.14;黏附系数为1.07±0.52;黏附指数为4.26±0.07。所研究的细菌具有高黏附活性。建立了生物膜光密度(D≥0.514 - 0.551)与平均黏附指数(AAI,4.15±0.28 - 4.76±0.75)之间的直接相关性(R = 0.94)。
本研究表明,[具体微生物名称未给出]具有高黏附活性,是生物膜的强力生产者,且样品的光密度超过对照光密度4倍以上。