全沟硬蜱围食膜初步观察研究

本工作用光镜和电镜技术对全沟硬蜱的围食膜进行了初步观察研究。结果表明，饥饿幼虫、若虫、成虫以及吸血期雄性成虫不具围食膜。吸血期幼虫、若虫及雌性成虫最晚分别于吸血开始后20、20及18小时出现了单层结构的围食膜；缓慢吸血期围食膜的厚度持续增加，而在快速吸血期其厚度却会显著减小。饱血后发育期围食膜不断增厚，愈来愈拱曲，与肠壁的距离不断扩大，并形成多层结构；至少分别于饱血后12、25及11天，仍能在幼虫、若虫及雌性成虫肠腔观察到完好状态的围食膜。全沟硬蜱是我国北方地区多种血液传播病原体的重要媒介，其围食膜的发现及围食膜形态变化的初步描述，对进一步认识这些病原体在全沟硬蜱体内迁移扩散规律、发育动态以及向宿主的传播途径，可提供有益的启示。     

二棘血蜱叮咬中华硬蜱纯化抗原免疫接种宿主后的交叉免疫反应

选用中华硬蜱105kD柱层析纯化抗原对新西兰兔进行免疫接种,再用二棘血蜱进行叮咬,并与单纯二棘血蜱叮咬组和佐剂对照组进行对比,以探讨不同种属硬蜱之间的交叉免疫抗性。结果显示:单纯二棘血蜱叮咬组吸血量为181 3±44 3mg,而二棘血蜱叮咬由中华硬蜱105kD纯化抗原免疫接种组和佐剂对照组新西兰兔后其吸血量分别为102 1±25 3mg和168 5±40 9mg,纯化抗原免疫接种组较单纯二棘血蜱叮咬组和佐剂对照组吸血量分别下降43 7%和39 4%(P<0 01)。同时,纯化抗原免疫接种组也较单纯二棘血蜱叮咬组和佐剂对照组产卵量有显著下降。该研究结果表明中华硬蜱和二棘血蜱之间存在着交叉免疫反应。     

Detection of Invasive Borrelia burgdorferi Strains in North‐Eastern Piedmont,Italy

Following reports of human cases of Lyme borreliosis from the Ossola Valley, a mountainous area of Piemonte, north‐western Italy, the abundance and altitudinal distribution of ticks, and infection of these vectors with Borrelia burgdorferi sensu lato were evaluated. A total of 1662 host‐seeking Ixodes ricinus were collected by dragging from April to September 2011 at locations between 400 and 1450 m above sea level. Additional 104 I. ricinus were collected from 35 hunted wild animals (4 chamois, 8 roe deer, 23 red deer). Tick density, expressed as the number of ticks per 100 m², resulted highly variable among different areas, ranging from 0 to 105 larvae and from 0 to 22 nymphs. A sample of 352 ticks (327 from dragging and 25 from wild animals) was screened by a PCR assay targeting a fragment of the 16S rRNA gene of B. burgdorferi s.l. Positive samples were confirmed with a PCR assay specific for the 5S‐23S rRNA intergenic spacer region and sequenced. Four genospecies were found: B. afzelii (prevalence 4.0%), B. lusitaniae (4.0%), B. garinii (1.5%) and B. valaisiana (0.3%). Phylogenetic analysis based on the ospC gene showed that most of the Borrelia strains from pathogenic genospecies had the potential for human infection and for invasion of secondary body sites.     

中华硬蜱唾液腺抗原成分的分析研究

中华硬蜱唾液腺提取物（ＳＧＥ）经ＳＤＳ聚丙烯酰胺凝胶电泳显示出２４条蛋白带，其中主带有６条，分子量分别为１４２、１０５、９４、６６、６４和５６ＫＤ。用中华硬蜱叮刺家兔后血清做免疫印渍，特异性抗原带为１０５和９４ＫＤ。说明中华硬蜱１０５和９４ＫＤ的蜱唾液腺蛋白原能够有效地诱慢宿主产生抗体。     

Retrospective study of 103 presumed cases of tick (Ixodes holocyclus) envenomation in the horse

Objective Review 103 cases of presumed tick envenomation in horses. Design Retrospective study. Method Variables, including date of presentation, age, breed, weight, presence of ticks, gait and respiration scores, duration of recumbency, treatment, outcome and complications were recorded. A series of univariable screening tests were performed and used in a multivariable logistic regression model. Results There were a total of 103 cases affecting 10 breeds, aged between 1 week and 18 years of age. Horses >6 months old and weighing >100 kg had a higher odds of death than those <6 months old and <100 kg. Cases were seen from North Queensland to the central coast of New South Wales and were more likely to present in the warmer months. There was no association between the number of ticks found on an animal and death. Horses with a higher respiratory score had higher odds of dying, but there was no association between gait score and survival. Horses recumbent >120 h after presentation had higher odds of dying. Complications were reported in 35% of horses. The odds ratio for survival was higher for horses receiving >0.5 mL/kg of tick antiserum. Overall, 74% of horses survived. Multivariable modelling was limited by the small sample size. Conclusion In general, tick envenomation in horses follows the geographic distribution of Ixodes holocyclus. Tick antiserum administered at >0.5 mL/kg increases the odds of survival. It would appear that the complications associated with managing a recumbent horse increase the odds of death.     

Following a tick bite: double infections by tick-borne encephalitis virus and the spirochete Borrelia and other potential multiple infections

In Central Europe and large parts of Asia, tick-borne-encephalitis (TBE) and Lyme borreliosis caused by the spirochetal bacterium of the genus Borrelia are among the most common diseases transmitted by the bite of a tick. When in regions with overlapping TBE virus and Borrelia endemicity, a tick bite causes the victim to become ill, it is important that appropriate serological and other laboratory investigations form part of the differential diagnosis. Account must always be taken of the fact that a tick bite may be followed by a double infection with the TBE virus and Borrelia. For this reason, a comprehensive diagnostic work-up aimed at detecting co-infection by both pathogens, even when the tick bite occurs in an endemic region for both pathogens but the initial clinical symptoms suggest an infection with only one of the two pathogens. The present article discusses a number of published cases of a co-infection with TBE virus and Borrelia and other potential multiple infections.     

Virus Detection in Questing Ticks is not a Sensitive Indicator for Risk Assessment of Tick‐Borne Encephalitis in Humans

Tick‐borne encephalitis virus (TBEV) is the most important tick‐transmitted arbovirus causing human disease in Europe, but information on its endemic occurrence varies between countries because of differences in surveillance systems. Objective data are necessary to ascertain the disease risk for vaccination recommendations and other public health interventions. In two independent, separately planned projects, we used real‐time RT‐PCR to detect TBE virus in questing ticks. In Poland, 32 sampling sites were selected in 10 administrative districts located in regions where sporadic TBE cases were reported. In Germany, 18 sampling sites were selected in two districts located in a region with high TBE incidence. Altogether, >16 000 ticks were tested by real‐time RT‐PCR, with no sample testing positive for TBEV. A systematic search for published studies on TBEV prevalence in ticks in Poland and Germany also suggested that testing large numbers of collected ticks could not consistently assure virus detection in known endemic foci. Although assignment of results to administrative regions is essential for TBE risk mapping, this was possible in only 10 (investigating 22 417 ticks) of 15 published studies (>50 000 ticks) identified. We conclude that the collection and screening of ticks by real‐time RT‐PCR cannot be recommended for assessment of human TBE risk. Alternative methods of environmental TBEV monitoring should be considered, such as serological monitoring of rodents or other wildlife.     

Modelling the Phenological Relationships of Questing Immature Ixodes Ricinus (Ixodidae) Using Temperature and NDVI Data

All active stages of the tick Ixodes ricinus were collected monthly at two sites in northern Spain between the years 2000 and 2007. We used percentile accumulation of the active stage in the environment to evaluate simple and coherent correlations between accumulation of the active stages of larvae and nymphs and medium‐resolution MODIS satellite‐derived information on the climate, including monthly and accumulated temperature and the Normalized Difference Vegetation Index (NDVI). This framework is not intended to predict the actual abundance of ticks in the field as a measure of the hazard to humans, but to provide a basic structure for addressing the phenology of the tick in its geographic range. We demonstrated that the accumulation of larval ticks in the active stage is a sigmoid function of the accumulated temperature from the beginning of the calendar year. We also demonstrated that the accumulated temperature necessary to recruit nymphs from the questing larval stage is a function of the changes in accumulated larvae and nymphs and the accumulated temperature and NDVI recorded by the Aqua sensor. The low p‐values obtained in the regressions confirmed that such recruitment can be calculated using time intervals to estimate, for example, the beginning of the questing period or the time of the year when a population peak can be expected. The comparison among predicted and actual accumulated temperatures between larvae and nymph recruitment had an averaged error of ±20 days in one complete year. The use of accumulated temperature and NDVI proposed in this study opens up the re‐evaluation of reports on the phenology of the tick in Europe. This framework is intended to evaluate the same correlations along the tick's range and predict its phenological patterns in areas of pathogen transmission risk for humans.     

Experimental in vitro transmission of Babesia sp. (EU1) by Ixodes ricinus

Babesia sp. (EU1), first characterized in 2003, has been implicated in human cases of babesiosis in Italy, Austria and Germany. It has been identified in roe deer and in its suspected tick vector, Ixodes ricinus, in several European countries. The aim of the present study was to validate the competence of I. ricinus as a vector of Babesia sp. (EU1) via experimental infections. For this purpose, a parasite strain isolated from roe deer was cloned in sheep erythrocytes. After experimental infections, parasite DNA was successfully amplified by PCR in both eggs and larvae originating from infected I. ricinus females and in the salivary glands of females exposed to Babesia sp. (EU1) as nymphs. We also demonstrate that infected females were able to transmit parasite DNA during a new blood meal. Together with previous epidemiological studies, these results validate I. ricinus as a competent vector for Babesia sp. (EU1).     

Detection and molecular characterization of Theileria sp. in fallow deer (Dama dama) and ticks from an Italian natural preserve

The prevalence of piroplasms in a closed population of fallow deer (Dama dama L.) living in the Italian preserve of “Bosco della Mesola” - Ferrara (Mesola wood) was investigated. Blood samples and ticks were collected from 62 fallow deer. On microscopic observation, 28 (45.0%) blood samples were positive for piroplasms while PCR provided evidence for piroplasms infection in 47 (75.8%) fallow deer. The 67 ticks, collected from positive and negative animals, were identified as Ixodesricinus L., 1758 (89.6%) and Haemaphysalisconcinna Koch, 1844 (10.4%). At the PCR, four samples of I. ricinus were positive for piroplasms. The sequences of the 18S rRNA gene from both blood and ticks were identical and showed high identity (99.6%) with Theileria sp. 3185/02 (DQ866842) and Theileria capreoli (AY726011) from roe deer. Interestingly, the phylogenetical analyses evidenced differences between the Theileria strain from Mesola wood and the ones isolated in fallow deer from other Italian areas.