Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Seestrasse 55, 16868 Wusterhausen, Germany.
Vet Parasitol. 2010 Oct 11;173(1-2):24-31. doi: 10.1016/j.vetpar.2010.06.009. Epub 2010 Jun 15.
Neosporosis is an important cause of reproductive failure in cattle worldwide. Two different abortion patterns associated with Neospora caninum infection have been observed in cattle herds: endemic and epidemic abortion outbreaks. The endemic pattern is characterized by an abortion problem in a herd persisting for several months or years, and is assumed to be caused by reactivation of a chronic infection. In epidemic outbreaks, abortions concentrate within a short period of time, most likely due to a recent point source exposure of naïve animals to N. caninum. The aim of the study was to characterize five N. caninum-associated epidemic abortion outbreaks in Germany by serological and molecular techniques, including a p38-avidity-ELISA and typing of N. caninum in clinical samples by multilocus-microsatellite analysis. DNA extracts from the brain of 18 N. caninum infected fetuses from epidemic abortion outbreaks were characterized using 10 N. caninum-microsatellite markers. Nested-PCR protocols were developed to amplify the marker regions MS1B, MS3, MS5, MS6A, MS6B, MS7, MS12 and MS21 from clinical samples for subsequent analysis by capillary electrophoresis. Microsatellites MS2 and MS10 were analyzed by previously reported sequencing techniques. Most dams which had aborted showed a low-avidity IgG response to the N. caninum p38-antigen, and in three of the five studied herds, the majority of the dams at risk, which had not aborted, had also low-avidity responses suggesting that infection with N. caninum had recently occurred in most animals. A common microsatellite pattern prevailed in all fetuses from each individual epidemic outbreak. This pattern was unique for each herd. Although the number of epidemic abortion outbreaks analyzed was limited, the observation of a common microsatellite pattern, accompanied by a low-avidity IgG response against N. caninum in the dams, supports the hypothesis of a recent infection from a common point source. The genetic diversity of N. caninum observed among these outbreaks may indicate that not a particular N. caninum genotype but the horizontal infection route determines the occurrence of epidemic abortions.
新生隐球菌病是全球牛群生殖失败的一个重要原因。在牛群中观察到两种与新生隐球菌感染相关的不同流产模式:地方性和流行性流产爆发。地方性模式的特点是牛群中的流产问题持续数月或数年,被认为是由慢性感染的再激活引起的。在流行爆发中,流产集中在短时间内,很可能是由于最近有一批新的动物接触了新生隐球菌。本研究的目的是通过血清学和分子技术对德国的五起新生隐球菌相关的流行流产爆发进行特征描述,包括 p38-亲和力 ELISA 和对临床样本中新生隐球菌的多基因座微卫星分析进行分型。从流行流产爆发中感染新生隐球菌的 18 个胎儿的脑组织中提取 DNA ,并使用 10 个新生隐球菌微卫星标记进行特征描述。开发巢式 PCR 方案从临床样本中扩增标记区域 MS1B、MS3、MS5、MS6A、MS6B、MS7、MS12 和 MS21,然后通过毛细管电泳进行分析。通过之前报道的测序技术分析微卫星 MS2 和 MS10。大多数流产的母畜对新生隐球菌 p38 抗原的 IgG 亲和力较低,在五个研究的牛群中,有三分之二的高危未流产的母畜也有低亲和力反应,表明大多数动物最近都感染了新生隐球菌。每个个体流行流产爆发的所有胎儿中都存在一种共同的微卫星模式。这种模式对每个牛群都是独特的。尽管分析的流行流产爆发数量有限,但观察到共同的微卫星模式,以及母畜对新生隐球菌的低亲和力 IgG 反应,支持了最近从共同的源头感染的假设。在这些爆发中观察到的新生隐球菌遗传多样性可能表明,不是特定的新生隐球菌基因型,而是水平感染途径决定了流行流产的发生。