Molecular Detection of Tick-Borne Pathogens in Humans with Tick Bites and Erythema Migrans,in the Netherlands

BackgroundTick-borne diseases are the most prevalent vector-borne diseases in Europe. Knowledge on the incidence and clinical presentation of other tick-borne diseases than Lyme borreliosis and tick-borne encephalitis is minimal, despite the high human exposure to these pathogens through tick bites. Using molecular detection techniques, the frequency of tick-borne infections after exposure through tick bites was estimated.MethodsTicks, blood samples and questionnaires on health status were collected from patients that visited their general practitioner with a tick bite or erythema migrans in 2007 and 2008. The presence of several tick-borne pathogens in 314 ticks and 626 blood samples of this cohort were analyzed using PCR-based methods. Using multivariate logistic regression, associations were explored between pathogens detected in blood and self-reported symptoms at enrolment and during a three-month follow-up period.ResultsHalf of the ticks removed from humans tested positive for Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, Rickettsia monacensis, Borrelia miyamotoi and several Babesia species. Among 92 Borrelia burgdorferi s. l. positive ticks, 33% carried another pathogen from a different genus. In blood of sixteen out of 626 persons with tick bites or erythema migrans, DNA was detected from Candidatus Neoehrlichia mikurensis (n = 7), Anaplasma phagocytophilum (n = 5), Babesia divergens (n = 3), Borrelia miyamotoi (n = 1) and Borrelia burgdorferi s. l. (n = 1). None of these sixteen individuals reported any overt symptoms that would indicate a corresponding illness during the three-month follow-up period. No associations were found between the presence of pathogen DNA in blood and; self-reported symptoms, with pathogen DNA in the corresponding ticks (n = 8), reported tick attachment duration, tick engorgement, or antibiotic treatment at enrolment.ConclusionsBased on molecular detection techniques, the probability of infection with a tick-borne pathogen other than Lyme spirochetes after a tick bite is roughly 2.4%, in the Netherlands. Similarly, among patients with erythema migrans, the probability of a co-infection with another tick-borne pathogen is approximately 2.7%. How often these infections cause disease symptoms or to what extend co-infections affect the course of Lyme borreliosis needs further investigations.     

Disease Risk in a Dynamic Environment: The Spread of Tick-Borne Pathogens in Minnesota,USA

EcoHealth - As humans and climate change alter the landscape, novel disease risk scenarios emerge. Understanding the complexities of pathogen emergence and subsequent spread as shaped by landscape...     

Influence of the Biotope on the Tick Infestation of Cattle and on the Tick-Borne Pathogen Repertoire of Cattle Ticks in Ethiopia

Background

The majority of vector-borne infections occur in the tropics, including Africa, but molecular eco-epidemiological studies are seldom reported from these regions. In particular, most previously published data on ticks in Ethiopia focus on species distribution, and only a few molecular studies on the occurrence of tick-borne pathogens or on ecological factors influencing these. The present study was undertaken to evaluate, if ticks collected from cattle in different Ethiopian biotopes harbour (had access to) different pathogens.

Methods

In South-Western Ethiopia 1032 hard ticks were removed from cattle grazing in three kinds of tick biotopes. DNA was individually extracted from one specimen of both sexes of each tick species per cattle. These samples were molecularly analysed for the presence of tick-borne pathogens.

Results

Amblyomma variegatum was significantly more abundant on mid highland, than on moist highland. Rhipicephalus decoloratus was absent from savannah lowland, where virtually only A. cohaerens was found. In the ticks Coxiella burnetii had the highest prevalence on savannah lowland. PCR positivity to Theileria spp. did not appear to depend on the biotope, but some genotypes were unique to certain tick species. Significantly more A. variegatum specimens were rickettsia-positive, than those of other tick species. The presence of rickettsiae (R. africae) appeared to be associated with mid highland in case of A. variegatum and A. cohaerens. The low level of haemoplasma positivity seemed to be equally distributed among the tick species, but was restricted to one biotope type.

Conclusions

The tick biotope, in which cattle are grazed, will influence not only the tick burden of these hosts, but also the spectrum of pathogens in their ticks. Thus, the presence of pathogens with alternative (non-tick-borne) transmission routes, with transstadial or with transovarial transmission by ticks appeared to be associated with the biotope type, with the tick species, or both, respectively.     

Crown Spread Patterns for Five Deciduous Broad-leaved Woody Species: Ecological Significance of the Retention Patterns of Larger Branches

Patterns of crown spread and branch retention of two shade-intolerantspecies (Betula platyphyllaandB. maximowicziana) were comparedwith three more tolerant species (Quercus mongolica,Acer sieboldianumandMagnoliaobovata). Branching height (height of the lowest living branch)rose more rapidly with age for the twoBetulaspecies than forthe shade tolerant species. Branching heights ofA. sieboldianumandM.obovatawere similar, irrespective of tree height and age, andlarger trees tended to produce wider crowns than theBetulaspecieswhen trees of similar height were compared. In all species,the branch basal area (cross-sectional area of a branch at itsbase) and the leaf area per branch generally increased as thebranch position on a stem became lower. Therefore, retaininglarger branches contributed significantly to the support ofa larger leaf area per tree. The number of larger branches (branchbasal area >80 cm²) for bothBetulaspecies was significantlysmaller than that of the shade tolerant species. The branchretention pattern ofBetulaspecies was probably a consequenceof intolerance of the leaves to shade. The decline ofBetulaspecieswith forest succession is likely to occur through their inabilityto retain branches with a large base area in closed forests.Copyright1997 Annals of Botany Company Shade tolerance; crown spread; branch retention; branch size; broad-leaved woody species; leaf area index per tree     

An Inhibitor of Thrombin-Stimulated Blood Platelet Aggregation from the Salivary Glands of the Hard Tick Amblyomma variegatum (Acari: Ixodidae)

The tropical bont tick, Amblyomma variegatum can cause intense skin irritation and inflammation and bites that often develop into septic wounds or abscess in their host. Crude salivary gland extract (SGE) of partially engorged A. variegatum females as well as SGE protein fractions purified by three-step reverse phase HPLC procedure were tested for their anti-aggregatory effect on isolated human blood platelets stimulated with thrombin and compared with the effect of recombinant hirudin. At concentrations 10⁻³ and 5 × 10⁻³ μg protein/ml the following rank order of antiplatelet activity was detected: AV 16/3 (inhibitor purified from AV-III, third purification) > SGE > AV-II (fraction from first purification) > AV-III (fraction from first purification) > hirudin. The effect of all fractions tested was dose-dependent. For fraction AV 16/3, the inhibitory effect was 49 and 61% for 10⁻³ and 5 × 10⁻³ μg protein/ml, respectively. The results suggest that protein fractions from A. variegatum SGE possess an antithrombin effect on human blood platelets with hirudin-like activity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mosaic Pattern of Borrelia Infection in a Continuous Population of the Tick Ixodes Ricinus (Acari: Ixodidae)

An array of 12 20 × 20 m quadrats in a mixed forest near Poteply, Central Bohemia, Czech Republic, was investigated for the abundance and spatial distribution of host-questing Ixodes ricinus nymphs and their infection with borreliae. Tick densities were estimated by flagging and their borrelia infection status was determined by direct immunofluorescence. While the tick population appeared to be continuous and homogeneous based on quadrat counts, their infection with borreliae evinced a mosaic pattern. The nymphal infection rates ranged between 1.6 and 10.5% (mean = 6.0%) with significant differences between adjacent quadrats. Home ranging of small rodents (Apodemus flavicollis and Clethrionomys glareolus) inhabiting the forest seems to be responsible for the spatial pattern of borrelial infection.     

Impact of the Infection Period Distribution on the Epidemic Spread in a Metapopulation Model

Epidemic models usually rely on the assumption of exponentially distributed sojourn times in infectious states. This is sometimes an acceptable approximation, but it is generally not realistic and it may influence the epidemic dynamics as it has already been shown in one population. Here, we explore the consequences of choosing constant or gamma-distributed infectious periods in a metapopulation context. For two coupled populations, we show that the probability of generating no secondary infections is the largest for most parameter values if the infectious period follows an exponential distribution, and we identify special cases where, inversely, the infection is more prone to extinction in early phases for constant infection durations. The impact of the infection duration distribution on the epidemic dynamics of many connected populations is studied by simulation and sensitivity analysis, taking into account the potential interactions with other factors. The analysis based on the average nonextinct epidemic trajectories shows that their sensitivity to the assumption on the infectious period distribution mostly depends on , the mean infection duration and the network structure. This study shows that the effect of assuming exponential distribution for infection periods instead of more realistic distributions varies with respect to the output of interest and to other factors. Ultimately it highlights the risk of misleading recommendations based on modelling results when models including exponential infection durations are used for practical purposes.     

Online Respondent-Driven Sampling for Studying Contact Patterns Relevant for the Spread of Close-Contact Pathogens: A Pilot Study in Thailand

Background

Information on social interactions is needed to understand the spread of airborne infections through a population. Previous studies mostly collected egocentric information of independent respondents with self-reported information about contacts. Respondent-driven sampling (RDS) is a sampling technique allowing respondents to recruit contacts from their social network. We explored the feasibility of webRDS for studying contact patterns relevant for the spread of respiratory pathogens.

Materials and Methods

We developed a webRDS system for facilitating and tracking recruitment by Facebook and email. One-day diary surveys were conducted by applying webRDS among a convenience sample of Thai students. Students were asked to record numbers of contacts at different settings and self-reported influenza-like-illness symptoms, and to recruit four contacts whom they had met in the previous week. Contacts were asked to do the same to create a network tree of socially connected individuals. Correlations between linked individuals were analysed to investigate assortativity within networks.

Results

We reached up to 6 waves of contacts of initial respondents, using only non-material incentives. Forty-four (23.0%) of the initially approached students recruited one or more contacts. In total 257 persons participated, of which 168 (65.4%) were recruited by others. Facebook was the most popular recruitment option (45.1%). Strong assortative mixing was seen by age, gender and education, indicating a tendency of respondents to connect to contacts with similar characteristics. Random mixing was seen by reported number of daily contacts.

Conclusions

Despite methodological challenges (e.g. clustering among respondents and their contacts), applying RDS provides new insights in mixing patterns relevant for close-contact infections in real-world networks. Such information increases our knowledge of the transmission of respiratory infections within populations and can be used to improve existing modelling approaches. It is worthwhile to further develop and explore webRDS for the detection of clusters of respiratory symptoms in social networks.     

节瓜蓟马空间分布图式研究

对节瓜蓟马在节瓜上的空间分布状况进行了调查和分析,结果表明,该虫的空间分布型前期符合奈曼分布,后期符合负二项分布;其空间图式是聚集的,分布的基本成分是个体群;在节瓜不同叶位上分布差异显著。明显向心叶聚集,在瓜栅向光面与背光面虫口比例方面,均以向光面较多。     

山东百部种群的空间分布格局

采用扩散系数的t检验、Poisson分布的x^2拟合检验及Morisita指数的F检验研究了山东百部（Stemona shandongensis）种群的空间分布类型;用负二项参数（K）、Cassie指标（1／K）、Green指数（GI）和Lloyd的平均拥挤度（m^＊）与聚块性指标（PAI）研究其聚集强度。同时利用上述指数研究了取样面积、苗龄对山东百部种群分布格局和聚集强度的影响,分析了分布格局形成的原因。结果表明,总体上山东百部种群的分布格局类型为集群分布,种群聚集尺度多为8m^2;当年生植株种群聚集强度高于多年生种群;整个种群分布格局的演化趋向于高聚集强度的集群分布。     

Interferon-Gamma and Nitric Oxide Synthase 2 Mediate the Aggregation of Resident Adherent Peritoneal Exudate Cells: Implications for the Host Response to Pathogens

Interferon-gamma (Ifnγ), a key macrophage activating cytokine, plays pleiotropic roles in host immunity. In this study, the ability of Ifnγ to induce the aggregation of resident mouse adherent peritoneal exudate cells (APECs), consisting primarily of macrophages, was investigated. Cell-cell interactions involve adhesion molecules and, upon addition of Ifnγ, CD11b re-localizes preferentially to the sites of interaction on APECs. A functional role of CD11b in enhancing aggregation is demonstrated using Reopro, a blocking reagent, and siRNA to Cd11b. Studies with NG-methyl-L-arginine (LNMA), an inhibitor of Nitric oxide synthase (Nos), NO donors, e.g., S-nitroso-N-acetyl-DL-penicillamine (SNAP) or Diethylenetriamine/nitric oxide adduct (DETA/NO), and Nos2 ^-/- mice identified Nitric oxide (NO) induced by Ifnγ as a key regulator of aggregation of APECs. Further studies with Nos2 ^-/- APECs revealed that some Ifnγ responses are independent of NO: induction of MHC class II and CD80. On the other hand, Nos2 derived NO is important for other functions: motility, phagocytosis, morphology and aggregation. Studies with cytoskeleton depolymerizing agents revealed that Ifnγ and NO mediate the cortical stabilization of Actin and Tubulin which contribute to aggregation of APECs. The biological relevance of aggregation of APECs was delineated using infection experiments with Salmonella Typhimurium (S. Typhimurium). APECs from orally infected, but not uninfected, mice produce high amounts of NO and aggregate upon ex vivo culture in a Nos2-dependent manner. Importantly, aggregated APECs induced by Ifnγ contain fewer intracellular S. Typhimurium compared to their single counterparts post infection. Further experiments with LNMA or Reopro revealed that both NO and CD11b are important for aggregation; in addition, NO is bactericidal. Overall, this study elucidates novel roles for Ifnγ and Nos2 in regulating Actin, Tubulin, CD11b, motility and morphology during the aggregation response of APECs. The implications of aggregation or “group behavior” of APECs are discussed in the context of host resistance to infectious organisms.     

Antibiotic Treatment in Mice Infected with Japanese Borrelia garinii: Efficacy of Ceftriaxone for Eradicating the Infection Induced by Ixodes persulcatus Tick Bites

We compared the efficacy of three antibiotics (ceftriaxone, erythromycin and clarithromycin) against Borrelia garinii infection in mice. The nymphal ticks of Ixodes persulcatus infected with the strain JEM6 of Japanese B. garinii were allowed to feed on female C3H mice. The mice were treated with each of the antibiotics for 5 consecutive days 1, 3, or 7 weeks after tick detachment. The doses of antibiotics per day were as follows: 5 mg intraperitoneal injection of ceftriaxone, 2 mg intraperitoneal injection of erythromycin and 1 mg peroral administration of clarithromycin. The infection status in treated mice was monitored by culturing their earlobes, hearts and urinary bladders in BSK II medium. Ceftriaxone eliminated borreliae completely; however, a recurrence of infection was observed in mice treated with erythromycin and clarithromycin.     

头部位移检测法动物悬尾实验仪的研制

采用红外传感、激光定位、51系列单片微机控制和记忆等技术,研制一种检测动物悬尾特性的仪器,用于药物研究中的动物悬尾实验。经30例小鼠实验证明,效果良好,该仪器为动物悬尾实验提供了一种全新的先进方法。     

The Known Distribution and Ecological Preferences of the Tick Subgenus Boophilus (Acari: Ixodidae) in Africa and Latin America

A compilation of the known distribution of Boophilus ticks in Africa and Latin America is presented, together with details on climate preferences. B. annulatus is recorded mainly in the western part of a strip from the equator to parallel 20° N. It associates with woodlands and forests (lowland rain forest and secondary grassland). This species is also present in the Mediterranean region, associated to woodland and open areas. B. decoloratus extends southern to parallel 20° N, in woodland with montane vegetation and Zambezian miombo; some records have been collected in the highveld grassland. B. geigyi is mainly collected in the western range of a stripe extending between parallels 5° N and 18° N, associated with Sudanian woodland, lowland rain forest with secondary grassland and woodland. Confirmed records of microplus in Africa are restricted to Malagasy region and south and eastern Africa, being predominant in the Zambezian miombo, deciduous forest with secondary grassland, and woodland. In Latin America, microplus is abundant in the Mesoamerican corridor to Venezuela and Colombia, and southern in Brazil and Argentina. The tick is mainly associated to the biomes of Chaco and Pampas in Argentina, the North-central moist Andes, the Atlantic forest (southern range) and the moist Meso-American vegetation (northern range). Most collections of B. annulatus and B. geigyi came from areas where winter minimum temperature is above 15 °C, maximum temperatures remain between 33 and 36 °C and maximum rainfall is recorded between June and September. B. decoloratus and African B. microplus are recorded in sites with low temperatures in May–September. Minimum temperature requirements are similar for both B. decoloratus and African B. microplus, and both are around 4 °C less than the value recorded for collections of Latin-American B. microplus. The rainfall pattern observed for decoloratus shows a minimum in May and June. The requirements of total rainfall are highest for B. microplus in Latin America, while records of African B. microplus are concentrated in areas of low rainfall between May and October, and high rainfall between November and March (low rainfall in the same period for B. decoloratus). Statistical analysis revealed the existence of populations (demes) with ecologically different requirements within each tick species. Both B. annulatus and B. decoloratus showed many different demes clearly associated to defined areas. The collections of Latin American B. microplus are very homogeneous according climate preferences and well separated from the African counterpart.     

Biosecurity and Vector Behaviour: Evaluating the Potential Threat Posed by Anglers and Canoeists as Pathways for the Spread of Invasive Non-Native Species and Pathogens

Invasive non-native species (INNS) endanger native biodiversity and are a major economic problem. The management of pathways to prevent their introduction and establishment is a key target in the Convention on Biological Diversity''s Aichi biodiversity targets for 2020. Freshwater environments are particularly susceptible to invasions as they are exposed to multiple introduction pathways, including non-native fish stocking and the release of boat ballast water. Since many freshwater INNS and aquatic pathogens can survive for several days in damp environments, there is potential for transport between water catchments on the equipment used by recreational anglers and canoeists. To quantify this biosecurity risk, we conducted an online questionnaire with 960 anglers and 599 canoeists to investigate their locations of activity, equipment used, and how frequently equipment was cleaned and/or dried after use. Anglers were also asked about their use and disposal of live bait. Our results indicate that 64% of anglers and 78.5% of canoeists use their equipment/boat in more than one catchment within a fortnight, the survival time of many of the INNS and pathogens considered in this study and that 12% of anglers and 50% of canoeists do so without either cleaning or drying their kit between uses. Furthermore, 8% of anglers and 28% of canoeists had used their equipment overseas without cleaning or drying it after each use which could facilitate both the introduction and secondary spread of INNS in the UK. Our results provide a baseline against which to evaluate the effectiveness of future biosecurity awareness campaigns, and identify groups to target with biosecurity awareness information. Our results also indicate that the biosecurity practices of these groups must improve to reduce the likelihood of inadvertently spreading INNS and pathogens through these activities.     

Effect of Rate of Photosynthesis on the Pattern of Assimilate Distribution in the Graminaceous Plant

Infrared gas analysis and a quantitative radiocarbon tracertechnique were used to measure photosynthesis and the distributionof ¹⁴C-labelled assimilate in Lolium temulentum and a uniculmbarley exposed continuously or intermittently to contrastinglight intensities. Plants grown for 10–20 d in low light(<50 W m^–2 of visible light) exported a greater proportionof their assimilate to growing leaves at the terminal meristemand a smaller proportion to their roots and tillers than equivalentplants in high light (150 W m^–2). A single day's exposure(8.4 h photoperiod) to a contrasting light regime elicited achange in the pattern of assimilate distribution in the samediurnal period, but 2–3 d exposure was required for asubstantial shift in the pattern of supply of assimilate tomeristems. The data indicated that in terms of assimilate distributioncomplete adaption to a new light regime is attained in about7 d.     

Effects of Heterogeneous Competitor Distribution and Ramet Aggregation on the Growth and Size Structure of a Clonal Plant

Spatially heterogeneous distribution of interspecific competitors and intraspecific aggregation of offspring ramets may affect the growth and size structure of clonal plant populations, but these have been rarely studied. We conducted a greenhouse experiment in which we grew a population of eight offspring ramets (plants) of the stoloniferous clonal plant Hydrocotyle vulgaris aggregately or segregately in two homogeneous treatments with or without a competing grass Festuca elata and a heterogeneous treatment with a patchy distribution of the grass. In patchy grass treatments, H. vulgaris produced markedly more biomass, ramets and stolons in open patches (without grasses) than in grass patches, but displayed lower size variations as measured by coefficient of variation of biomass, ramets and stolons among the eight plants. In open areas, H. vulgaris produced statistically the same amounts of biomass and even more stolons and showed higher size variations in patchy grass treatments than in open (no grass) treatments. In grass areas, H. vulgaris grew much worse and displayed higher size variations in patchy grass treatments than in full grass treatments. Ramet aggregation decreased the growth of H. vulgaris in open treatments and in both open and grass patches in patchy grass treatments, but had little effect in full grass treatments. Ramet aggregation had little effect on size variations. Therefore, heterogeneous distribution of competitors can affect the growth and size structure of clonal plant populations, and ramet aggregation may decrease population growth when they grow in open environments or heterogeneous environments with a patchy distribution of interspecific competitors.     

孑遗植物裸果木历史分布格局模拟及避难区研究

裸果木(Gymnocarpos przewalskii)是国家一级保护植物,亦是古地中海第三纪孑遗成分。该文基于Maxent模型结合ArcGIS预测裸果木种群自冰期以来的分布变化、影响种群分布的主导气候因子及气候避难所位置。结果表明:随历史气候的变迁裸果木由末次间冰期经末次盛冰期进入全新中期,分布区面积呈现显著下降的趋势,而适应区面积呈先降低后增加趋势。目前主要分布于新疆中西部、甘肃河西走廊中部、玉门关以西北缘、内蒙古西部及宁夏北部等地。因祁连山山脉、阿尔金山山脉、昆仑山山脉及贺兰山山脉的阻挡,冰期在甘肃的河西走廊(疏勒河、党河流域)、玉门关西北缘(哈密)、内蒙古西部(雅布赖)及新疆(塔里木)等盆地形成了部分避难区。采用刀切法筛选出影响种群分布变化的4个主导因子及变化区间,即年降雨量变化范围为64.49~116.36mm、最湿季度降雨量变化范围为35.99~72.82mm、最冷季节温度变化范围为-13.31~-8.18℃及年均温变化范围为6.20~7.31℃。相对于温度的影响,水分对种群分布的影响更大。