Wittenbrink M M, Reuter C, Manz M L, Krauss H
Institut für Hygiene und Infektionskrankheiten der Tiere, Fachbereich Veterinärmedizin, Justus-Liebig-Universität Giessen, Germany.
Zentralbl Bakteriol. 1996 Sep;285(1):20-8. doi: 10.1016/s0934-8840(96)80018-1.
A total of 971 Ixodes (I.) ricinus ticks field-collected in May and June 1993 and 1994 in two different geographic regions (Middle Hesse and Lower Saxony) were examined for the presence of Lyme borreliosis spirochetes (Borrelia burgdorferi sensu lato) by primary culture using nonselective and selective BSK II medium. Modifications of BSK II medium with varying concentrations of bovine serum albumin and with rabbit serum being replaced by horse serum were examined for their ability to support growth of borreliae from ticks. Spirochetes were isolated from 17.8% each of female and male I. ricinus from Lower Saxony. Isolation rates from female and male I. ricinus originating from Middle Hesse were 13.5% and 19.6%, respectively. In comparison to nonselective BSK II medium the isolation rate of borreliae from the tick midgut was increased significantly (35%) by additional culture in selective BSK II medium. Most spirochetal isolates (85%) were detected within three weeks of incubation. Compared to isolation in standard BSK II medium, reduction of bovine serum albumin by 50% did not diminish the isolation rate of borreliae from ticks. Spirochetal isolates were identified as Borrelia burgdorferi sensu lato by PCR using a primer set based on ribosomal DNA sequences to amplify the variable spacer region between two conserved structures, the 3'end of the 5S rRNA and the 5'end of the 23S rRNA.
1993年5月和6月以及1994年5月和6月在两个不同地理区域（黑森州中部和下萨克森州）野外采集的971只蓖麻硬蜱，使用非选择性和选择性BSK II培养基通过原代培养检测莱姆病螺旋体（广义伯氏疏螺旋体）的存在。研究了用不同浓度牛血清白蛋白修饰的BSK II培养基以及用马血清替代兔血清的培养基支持蜱中疏螺旋体生长的能力。在下萨克森州的雌性和雄性蓖麻硬蜱中，分别有17.8%分离出螺旋体。源自黑森州中部的雌性和雄性蓖麻硬蜱的分离率分别为13.5%和19.6%。与非选择性BSK II培养基相比，在选择性BSK II培养基中额外培养可使蜱中肠疏螺旋体的分离率显著提高（35%）。大多数螺旋体分离株（85%）在孵育三周内被检测到。与在标准BSK II培养基中分离相比，牛血清白蛋白减少50%并没有降低蜱中疏螺旋体的分离率。使用基于核糖体DNA序列的引物对通过PCR将螺旋体分离株鉴定为广义伯氏疏螺旋体，以扩增两个保守结构（5S rRNA的3'端和23S rRNA的5'端）之间的可变间隔区。
