Nymphal Ticks Research Articles

Amblyomma hebraeum is the predominant tick species on goats in the Mnisi Community Area of Mpumalanga Province South Africa and is co-infected with Ehrlichia ruminantium and Rickettsia africae

BackgroundIn sub-Saharan Africa, Amblyomma ticks are vectors of heartwater disease in domestic ruminants, caused by the rickettsial pathogen Ehrlichia ruminantium. Immature tick stages often bite humans, whereby they act as vectors of tick-bite fever caused by Rickettsia africae. Moreover, Amblyomma ticks cause damage to livestock due to their feeding behaviour. In South Africa, we studied the abundance of Amblyomma hebraeum ticks on goats of emerging farmers in Mpumalanga Province. A selected number of A. hebraeum nymphs and adult ticks was tested for co-infection with E. ruminantium and R. africae.MethodsA total of 630 indigenous goats, belonging to farmers in the Mnisi Community area, were examined for ticks in 2013 and 2014. All ticks were identified, and a selected number was tested by PCR with reverse line blot hybridisation.ResultsIn total, 13,132 ticks were collected from goats distributed over 17 different households. Amblyomma hebraeum was the predominant species, followed by R. microplus. Rhipicephalus appendiculatus, R. simus and R. zambeziensis were also identified. Amblyomma hebraeum was present throughout the year, with peak activity of adults in summer (November) and nymphs in winter (July). The ratio between adults and nymphs ranged from 1:2.7 in summer to 1:55.1 in winter. The mean prevalence of infection for E. ruminantium by PCR/RLB in adult ticks was 17.4% (31/178), whereas 15.7% (28/178) were infected with R. africae. In pooled nymphs, 28.4% were infected with E. ruminantium and 38.8% carried R. africae infection. Co-infections of E. ruminantium and R. africae in adult and pooled nymphal ticks were 3.9% (7/178) and 10% (14.9), respectively. Lameness of goats due to predilection of ticks for the interdigital space of their feet was observed in 89% of the households.ConclusionsGoats act as important alternative hosts for cattle ticks, which underscored the necessity to include goats in control programs. It is suggested to use acaricide-impregnated leg-bands as a sustainable method to kill ticks and prevent lameness in goats. The challenge of goats by considerable numbers of E. ruminantium-infected ticks is a major obstacle for upgrading the indigenous goat breeds. Humans may be at risk to contract tick-bite fever in this area.

Monitoring of Nesting Songbirds Detects Established Population of Blacklegged Ticks and Associated Lyme Disease Endemic Area in Canada.

This study provides a novel method of documenting established populations of bird-feeding ticks. Single populations of the blacklegged tick, Ixodes scapularis, and the rabbit tick, Haemaphysalis leporispalustris, were revealed in southwestern Québec, Canada. Blacklegged tick nymphs and, similarly, larval and nymphal rabbit ticks were tested for the Lyme disease bacterium, Borrelia burgdorferi sensu lato (Bbsl), using PCR and the flagellin (flaB) gene, and 14 (42%) of 33 of blacklegged tick nymphs tested were positive. In contrast, larval and nymphal H. leporsipalustris ticks were negative for Bbsl. The occurrence of Bbsl in I. scapularis nymphs brings to light the presence of a Lyme disease endemic area at this songbird nesting locality. Because our findings denote that this area is a Lyme disease endemic area, and I. scapularis is a human-biting tick, local residents and outdoor workers must take preventive measures to avoid tick bites. Furthermore, local healthcare practitioners must include Lyme disease in their differential diagnosis.

The distribution limit of the common tick, Ixodes ricinus, and some associated pathogens in north-western Europe

In north-western Europe, the common tick, Ixodes ricinus, is widely established, its distribution appears to be increasing and the spread of tick-borne diseases is of increasing concern. The project ‘Flått i Nord’ (Ticks in northern Norway) commenced in spring 2009 with the intention of studying the tick’s distribution and that of its pathogens in northern Norway. Several methods were used: cloth-dragging, collecting from trapped small mammals, and collecting from pets. Since 2010, the occurrence of ticks in the region of northern Norway was determined directly by cloth-dragging 167 times in 109 separate locations between the latitudes of 64 °N and 70 °N (included seven locations in the northern part of Trøndelag County). The northernmost location of a permanent I. ricinus population was found to be Nordøyvågen (66.2204 °N, 12.59 °E) on the Island of Dønna. In a sample of 518 nymphal and adult ticks, the Borrelia prevalence collected close to this distribution limit varied but was low (1–15 %) compared with the locations in Trøndelag, south of the study area (15–27 %). Five specimens (1 %) were positive for Rickettsia helvetica. The length of the vegetation growing season (GSL) can be used as an approximate index for the presence of established populations of I. ricinus. The present study suggests that the threshold GSL for tick establishment is about 170 days, because the median GSL from 1991 to 2015 was 174–184 days at sites with permanent tick populations, showing a clear increase compared with the period 1961–1990. This apparent manifestation of climate change could explain the northward extension of the range of I. ricinus.

Identification of bacteria in the Rocky Mountain wood tick, Dermacentor andersoni, using single-strand conformation polymorphism (SSCP) and DNA sequencing.

PCR-based single-strand conformation polymorphism (SSCP) analyses combined with DNA sequencing of the prokaryotic 16S ribosomal (r) RNA gene encompassing the hypervariable V4 region was used to determine the bacterial composition of Rocky Mountain wood ticks (Dermacentor andersoni) attached to Richardson's ground squirrels (Urocitellus richardsonii) and questing on vegetation in southern Saskatchewan, Canada. The bacteria present in questing adult ticks from Saskatchewan Landing Provincial Park included Rickettsia peacockii, a Francisella-like endosymbiont (FLE) and an Arsenophonus-like endosymbiont. Bacteria in the adult and nymphal ticks attached to U. richardsonii collected from Beechy included R. peacockii, a FLE, and several other genera (e.g., Ralstonia, Sphingobium, Comamonas and Pseudomonas). The bacteria detected in D. andersoni in the present study are consistent with the findings of other studies that have characterized the microbiome of this tick species in the USA using next generation sequencing. This result demonstrates that the SSCP-based approach used in this study is cost- and time-effective for examining bacterial composition in ticks.

The potential impact of climate change on the transmission risk of tick-borne encephalitis in Hungary

BackgroundImpact of climate change on tick-borne encephalitis (TBE) prevalence in the tick-host enzootic cycle in a given region depends on how the region-specific climate change patterns influence tick population development processes and tick-borne encephalitis virus (TBEV) transmission dynamics involving both systemic and co-feeding transmission routes. Predicting the transmission risk of TBEV in the enzootic cycle with projected climate conditions is essential for planning public health interventions including vaccination programs to mitigate the TBE incidence in the inhabitants and travelers. We have previously developed and validated a mathematical model for retroactive analysis of weather fluctuation on TBE prevalence in Hungary, and we aim to show in this research that this model provides an effective tool for projecting TBEV transmission risk in the enzootic cycle.MethodsUsing the established model of TBEV transmission and the climate predictions of the Vas county in western Hungary in 2021-2050 and 2071-2100, we quantify the risk of TBEV transmission using a series of summative indices - the basic reproduction number, the duration of infestation, the stage-specific tick densities, and the accumulated (tick) infections due to co-feeding transmission. We also measure the significance of co-feeding transmission by observing the cumulative number of new transmissions through the non-systemic transmission route.ResultsThe transmission potential and the risk in the study site are expected to increase along with the increase of the temperature in 2021-2050 and 2071-2100. This increase will be facilitated by the expected extension of the tick questing season and the increase of the numbers of susceptible ticks (larval and nymphal) and the number of infected nymphal ticks co-feeding on the same hosts, leading to compounded increase of infections through the non-systemic transmission.ConclusionsThe developed mathematical model provides an effective tool for predicting TBE prevalence in the tick-host enzootic cycle, by integrating climate projection with emerging knowledge about the region-specific tick ecological and pathogen enzootic processes (through model parametrization fitting to historical data). Model projects increasing co-feeding transmission and prevalence of TBEV in a recognized TBE endemic region, so human risk of TBEV infection is likely increasing unless public health interventions are enhanced.

Newly identified viral genomes in pangolins with fatal disease.

Epizootic pathogens pose a major threat to many wildlife species, particularly in the context of rapidly changing environments. Pangolins (order Pholidota) are highly threatened mammals, in large part due to the trade in illegal wildlife. During July to August 2018 four sick wild pangolins (three Manis javanica and one Manis pentadactyla) exhibiting a variety of clinical symptoms were rescued by the Jinhua Wildlife Protection Station in Zhejiang province, China. Although three of these animals died, fortunately one recovered after 2 weeks of symptomatic treatment. Using meta-transcriptomics combined with reverse transcription polymerase chain reaction (RT-PCR), we identified two novel RNA viruses in two of the dead pangolins. Genomic analysis revealed that these viruses were most closely related to pestiviruses and coltiviruses, although still highly genetically distinct, with more than 48 and 25 per cent sequence divergence at the amino acid level, respectively. We named these Dongyang pangolin virus (DYPV) and Lishui pangolin virus (LSPV) based on the sampling site and hosts. Although coltiviruses (LSPV) are known to be transmitted by ticks, we found no evidence of LSPV in ticks sampled close to where the pangolins were collected. In addition, although DYPV was present in nymph ticks (Amblyomma javanense) collected from a diseased pangolin, they were not found in the local tick population. Epidemiological investigation revealed that both novel viruses might have been imported following the illegal international trade of pangolins. Hence, these data indicate that illegal wildlife trafficking not only threatens the status of pangolin populations, but may also spread epizootic pathogens.

Auto-dissemination of commercially available fungal pathogens in a laboratory assay for management of the brown dog tick Rhipicephalus sanguineus.

Rhipicephalus sanguineus is a canine tick that infests dogs throughout the world and is frequently found in homes and dog kennels. Management of this tick species is complicated by the presence of resistance to commonly utilized acaricides. Fungal formulations could provide a valuable alternative tool for management and are especially relevant indoors where detrimental environmental effects on fungal spores are of less concern. Two commercially available fungal formulations, one containing Metarhizium anisopliae and the other containing Beauveria bassiana, were compared for time to death and sporulation in nymphal ticks exposed for 60 min in treated filter paper packets. Beauveria bassiana exposure killed ticks faster than M. anisopliae exposure and B. bassiana was more likely to sporulate on tick cadavers than M. anisopliae. To determine whether infected ticks could disseminate fungus to their conspecifics, ticks were marked and treated with fungus before being placed with untreated ticks. Fungus was successfully transmitted from treated to untreated ticks. Mortality of ticks exposed to B. bassiana-exposed conspecifics occurred sooner than for those exposed to M. anisopliae-exposed conspecifics, indicating faster dissemination in the former. Therefore, although both formulations resulted in decreased longevity of ticks compared with the controls, the B. bassiana formulation holds the most promise for direct or indirect application with respect to brown dog tick management.

Woodchip borders at the forest ecotone as an environmental control measure to reduce questing tick density along recreational trails in Ottawa, Canada

Recent studies have highlighted the occurrence and distribution of Ixodes scapularis ticks infected with Borrelia burgdorferi sensu stricto (s.s.) around the city of Ottawa, Ontario, Canada, and the need for strategies to mitigate the risk of human exposure and infection. We conducted a field study from July to October 2018 to examine the effectiveness of ecotonal woodchip borders as an environmental control method to suppress the density of host-seeking ticks along recreational trails in Ottawa. We used an experimental design with ten 100-m trail replicates randomized to intervention or control groups, and monitored questing tick density at weekly intervals in mid-summer and early fall. We compared questing tick density between woodchip-treated and untreated trails using a mixed-effects Poisson regression model. Of the 138 I. scapularis ticks collected, there were 86 adult and nymphal ticks, 37 (43 %) of which were positive for B. burgdorferi s.s. A total of 58 Haemaphysalis leporispalustris were also collected. Mean combined adult and nymphal I. scapularis density was 1.15 (1.40 standard deviation; SD) per 100 m in the control group compared to 0.28 (0.56 SD) per 100 m in the intervention group, reflecting a 75 % reduction in questing tick density on trail replicates treated with woodchip borders (p < 0.001). An effect of the intervention was observed in both sampling periods. This study indicates that woodchip borders may be an effective strategy to suppress questing tick density along trail margins where recreational trail users are more active, thereby reducing the likelihood of tick encounters.

First evidence of Ixodiphagus hookeri (Hymenoptera: Encyrtidae) parasitization in Finnish castor bean ticks (Ixodes ricinus)

Ixodiphagus hookeri (Hymenoptera: Encyrtidae) is a parasitoid wasp specialized in parasitizing the larvae and nymphs of ticks (Acari: Ixodidae). As parasitized ticks die prior to reproduction, I. hookeri is seen as a prime biological control agent candidate. Despite this, little is known of their occurrence or ecology in northern Europe. The main aim of the current study was to determine whether adult wasps or parasitized ticks can be found from a tick-infested island in southwestern Finland, using field collections and molecular methods. Following the initial discovery of an adult I. hookeri female on Seili Island, we set out to collect further specimens via sweep netting and Malaise trappings between May and October 2017. Furthermore, 1310 Ixodes ricinus (1220 nymphs, 90 adults) collected from the island during 2012–2014 were screened for I. hookeri DNA using qPCR. Whereas no further wasp specimens could be collected via sweep netting or Malaise trappings, I. hookeri DNA was consistently detected in I. ricinus nymphs (annual minimum infection rates in 2012, 2013, and 2014: 2.3, 0.4, and 0.5%, respectively), whereas all adult samples were negative. Although the annually repeated detections of parasitized ticks suggest that the wasp inhabits the island, further field and molecular surveys are needed to more comprehensively determine the status and stability of the population.

Incorporating tick feeding behaviour into [formula omitted] for tick-borne pathogens

Tick-borne pathogens pose a considerable disease burden in Europe and North America, where increasing numbers of human cases and the emergence of new tick-borne pathogens has renewed interest in resolving the mechanisms underpinning their geographical distribution and abundance. For Borrelia burgdorferi and tick-borne encephalitis (TBE) virus, transmission of infection from one generation of ticks to another occurs when older nymphal ticks infect younger larval ticks feeding on the same host, either indirectly via systemic infection of the vertebrate host or directly when feeding in close proximity. Here, expressions for the basic reproduction number, R0, and the related tick type-reproduction number, T, are derived that account for the observation that larval and nymphal ticks tend to aggregate on the same minority of hosts, a tick feeding behaviour known as co-aggregation. The pattern of tick blood meals is represented as a directed, acyclic, bipartite contact network, with individual vertebrate hosts having in-degree, kin, and out-degree, kout, that respectively represent cumulative counts of nymphal and larval ticks fed over the lifetime of the host. The in- and out-degree are not independent when co-aggregation occurs such that T∝〈kinkout〉〈kin〉,where 〈.〉 indicates expected value. When systemic infection in the vertebrate host is the dominant transmission route R02=T, whereas when direct transmission between ticks co-feeding on the same host is dominant then R0=T and the effect of co-aggregation on R0 is more pronounced. Simulations of B. burgdorferi and TBE virus transmission on theoretical tick-mouse contact networks revealed that aggregation and co-aggregation have a synergistic effect on R0 and T, that co-aggregation always increases R0 and T, and that aggregation only increases R0 and T when larvae and nymphs also co-aggregate. Co-aggregation has the greatest absolute effect on R0 and T when the mean larval burden of hosts is high, and the largest relative effect on R0 for pathogens sustained by co-feeding transmission, e.g. TBE virus in Europe, compared with those predominantly spread by systemic infection, e.g. B. burgdorferi. For both pathogens, though, co-aggregation increases the mean number of ticks infected per infectious tick, T, and so too the likelihood of pathogen persistence.

1614. Single-Dose Doxycycline as Lyme Disease Post-Exposure Prophylaxis in a National Commercial Insurance Claims Database―the United States, 2014–2017

BackgroundApproximately 300,000 cases of Lyme disease occur annually in the UNITED STATES, with children aged 5–9 years disproportionately affected. A single dose of doxycycline administered within 72 hours of a high-risk tick bite is recommended for post-exposure prophylaxis (PEP) to prevent Lyme disease in areas of high incidence. However, it is not known how often or for which patients PEP is used. We aimed to describe recent patterns of single-dose doxycycline medication claims in states with high and low Lyme disease incidence, and the associated patient and prescription characteristics in a large national commercial insurance claims database.MethodsOutpatient medication claims in the IBM Watson Health MarketScan Database®, a large nation-wide database of de-identified insurance claims filed between January 1, 2014–December 31, 2017 were reviewed. Claims of single-dose doxycycline were identified and associated patient demographics and medication characteristics were analyzed.ResultsDuring 2014–2017, 66,210 medication claims for single-dose doxycycline were filed by 63,112 enrollees; mean annual incidence of receiving at least one single-dose doxycycline prescription was 56 per 100,000 enrollees. Mean patient age was 43 years (IQR 33–56 years); only 8% were for children aged <18 years. About half (46%) were male patients. Most claims (71%) were made by patients residing in the 14 states with high Lyme disease incidence, defined as an average annual incidence of ≥ 10 confirmed Lyme disease cases per 100,000 population. The majority (80%) of medication claims were during the 6 months of peak tick activity (April–July for nymphal ticks and October–November for adult ticks).ConclusionSingle-dose doxycycline medication claims are common in states with high Lyme disease incidence and are highest during months of peak tick activity, consistent with the assumption that most single-dose doxycycline is used for Lyme disease PEP. Use of single-dose doxycycline to prevent Lyme disease is infrequent in children, despite being a group at high risk for Lyme disease. Efforts to educate pediatric healthcare providers and parents should be made to increase Lyme disease PEP access for children.Disclosures All authors: No reported disclosures.

How the Distance Between Drag-Cloth Checks Affects the Estimate of Adult and Nymphal Ixodes scapularis (Acari: Ixodidae) Density.

Drag-cloth sampling is the most commonly used method to sample for ticks. A cloth is dragged along the ground and checked for ticks at regular intervals to count ticks before they drop off. The distance between drag-cloth checks differs between studies, which could result in lower density estimates for studies with greater distances between checks. Here, we measured this effect by 1) calculating the rate at which nymphal and adult Ixodes scapularis Say ticks drop off the cloth per meter dragged and 2) measuring tick density by drag-cloth sampling with three different drag-cloth check interval distances. We found a higher drop-off rate for adult ticks, 0.083/m, than nymphal ticks, 0.047/m. The estimated density of ticks decreased with increasing check interval distance. Our results not only highlight the importance of accounting for check interval distance when estimating tick density, but also provide the first estimate of nymphal I. scapularis drop-off rate.

Habitat amount, quality, and fragmentation associated with prevalence of the tick-borne pathogen Ehrlichia chaffeensis and occupancy dynamics of its vector, Amblyomma americanum

Tick-borne diseases are becoming increasingly prevalent world-wide. This is likely due in part to land-cover change, particularly forest fragmentation, but this evidence is largely limited to Lyme disease. It is unknown whether this is generalizable to other, emergent tick-borne pathogens. Motivated by hypotheses regarding landscape context and vertebrate hosts, we asked how landscape context, namely habitat amount, quality, and fragmentation, is related to the distribution of Ehrlichia chaffeensis, a tick-borne pathogen of increasing concern, and the interannual occupancy dynamics of its vector, the lone star tick (Amblyomma americanum). We collected nymphal ticks from 130 plots in southeastern Virginia, U.S., for 5 years and tested for E. chaffeensis via targeted PCR. We derived metrics of landscape context from Landsat data and related these to pathogen prevalence and tick turnover using hierarchical Bayesian models. Landscape context was associated with both pathogen prevalence and tick turnover. Pathogen prevalence was negatively associated with total forest landcover, coniferous forest landcover, and forest edge density. Tick turnover was positively associated with coniferous landcover and with an interaction between total forest landcover and edge. This interaction was such that turnover was predicted to be lowest in small contiguous forests, and highest in small fragmented forests. Landscape context affects E. chaffeensis prevalence and occupancy dynamics of its tick host, though these processes appear decoupled. We hypothesize that pathogen prevalence may be more driven by reservoir host movement and social behavior and tick dynamics are more driven by host population density.

Evaluation of NEON Data to Model Spatio-Temporal Tick Dynamics in Florida.

In 2013, the National Ecological Observatory Network (NEON) started collecting 30-year multi-faceted ecological data at various spatial and temporal scales across the US including ticks. Understanding the abundance and dynamics of disease vectors under changing environmental conditions in the long-term is important to societies, but sustained long-term collection efforts are sparse. Using hard-bodied tick data collected by NEON, the vegetation and atmospheric data and a statistical state-space model, which included a detection probability component, this study estimated the abundance of tick nymphs and adult ticks across a Florida NEON location. It took into account the spatial and temporal variation, and factors affecting detection. Its purpose was to test the applicability of data collected thus far and evaluate tick abundance. The study found an increase in tick abundance at this Florida location, and was able to explain spatial and temporal variability in abundance and detection. This approach shows the potential of NEON data. The NEON data collection is unique in scale, and promises to be of great value to understand tick and disease dynamics across the US. From a public health perspective, the detection probability of vectors can be interpreted as the probability of encountering that vector, making these types of analyses useful for estimating disease risk.

Impacts of Deciduous Leaf Litter and Snow Presence on Nymphal Ixodes scapularis (Acari: Ixodidae) Overwintering Survival in Coastal New England, USA.

Blacklegged ticks (Ixodes scapularis Say) are the vector for pathogens that cause more cases of human disease than any other arthropod. Lyme disease is the most common, caused by the bacterial spirochete Borrelia burgdorferi (Johnson, Schmid, Hyde, Steigerwalt, and Brenner) in the northeastern United States. Further knowledge of seasonal effects on survival is important for management and modeling of both blacklegged ticks and tick-borne diseases. The focus of our study was on the impact of environmental factors on overwintering success of nymphal blacklegged ticks. In a three-year field study conducted in Connecticut and Maine, we determined that ground-level conditions play an important role in unfed nymphal overwintering survival. Ticks in plots where leaf litter and snow accumulation were unmanipulated had significantly greater survival compared to those where leaf litter was removed (p = 0.045) and where both leaf litter and snow were removed (p = 0.008). Additionally, we determined that the key overwintering predictors for nymphal blacklegged tick survival were the mean and mean minimum temperatures within a year. The findings of this research can be utilized in both small- and large-scale management of blacklegged ticks to potentially reduce the risk and occurrence of tick-borne diseases.

Epidemiological Investigation of Tick Species from Near Domestic Animal Farms and Cattle, Goat, and Wild Boar in Korea.

This study aimed to investigate the tick species and give background for tick-borne investigations in Korea. Ticks were collected from the area within 2 km radius of the 4 domestic animal farms, where they were located in mountainous areas and raising animals on pasture, and from animal bodies in 2014 and 2015. In total, 7,973 nymphal and adult ticks were collected from the farms - 7,758 Haemaphysalis longicornis, 198 Haemaphysalis flava, and 17 Ixodes nipponensis, and 1,763 were collected from animals - 729 H. longicornis from cattle; 569 H. longicornis from goats; and 297 H. longicornis, 118 H. flava, 1 I. nipponensis, and 49 Amblyomma testudinarium from wild boars. As more species of ticks were collected from wild boars than domesticated animals and their habitats, various animal hosts should be considered while investigating tick species.

Epidemiology of SFTSV in Ticks collected from National Park in the ROK, 2015-2018

ObjectiveTo survey the distribution of Ixodid tick and infection of severe fever with thrombocytopenia syndrome virus (SFTSV) in natural environments from Deogyusan national parks in Korea.IntroductionSevere fever with thrombocytopenia syndrome (SFTS) is an emerging viral disease in East-Asian countries, including China, Japan, and the Republic of Korea (ROK). The causative agent of SFTS is the SFTS virus (SFTSV) transmitted by hematophagous ticks.MethodsTo investigate the prevalence of SFTSV in the ROK, a total of 4,223 ticks were collected by flagging from Deogyusan National Park from 2015 to 2018. One-step reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR were used to detect SFTSV-specific gene fragment from each ticks. The sequence data were analyzed using Chromas and aligned using CLUSTAL X. The phylogenetic analysis was constructed using the neighbor-joining method in MEGA7.ResultsOf the collected adult and nymph ticks, Haemaphysalis longicornis (3611, 85.5%) were the most abundant, followed by H. flava (502, 11.88%), Ixodes nipponensis (109, 2.5%), and Ixodes ovatus (1, 0.02%). The infection rate of SFTSV in total ticks was 5.8% (245/4,223), and the infection rate by year was 3.69% in 2015, 7.97% in 2016, 5.08% in 2017 and 4.68% in 2018. The infection rates of SFTSV were getting decreased each year in Deogyusan National Park. In addition, infection rate was higher in spring and summer of each season. Phylogenetic analysis was performed and SFTSV sequences obtained in this study were included in Korean/Japanese SFTSV clade.ConclusionsIn conclusion, we confirmed the sequence of two clades, and it is thought that the epidemiological investigation of SFTSV is necessary through further studies.

Lyme neuroborreliosis and bird populations in northern Europe.

Many vector-borne diseases are transmitted through complex pathogen-vector-host networks, which makes it challenging to identify the role of specific host groups in disease emergence. Lyme borreliosis in humans is now the most common vector-borne zoonosis in the Northern Hemisphere. The disease is caused by multiple genospecies of Borrelia burgdorferi sensu lato bacteria transmitted by ixodid (hard) ticks, and the major host groups transmit Borrelia genospecies with different pathogenicity, causing variable clinical symptoms in humans. The health impact of a given host group is a function of the number of ticks it infects as well as the pathogenicity of the genospecies it carries. Borrelia afzelii, with mainly small mammals as reservoirs, is the most common pathogen causing Lyme borreliosis, and it is often responsible for the largest proportion of infected host-seeking tick nymphs in Europe. The bird-borne Borrelia garinii, though less prevalent in nymphal ticks, is more likely to cause Lyme neuroborreliosis, but whether B. garinii causes disseminated disease more frequently has not been documented. Based on extensive data of annual disease incidence across Norway from 1995 to 2017, we show here that 69% of disseminated Lyme borreliosis cases were neuroborreliosis, which is three times higher than predicted from the infection prevalence of B. garinii in host-seeking ticks (21%). The population estimate of migratory birds, mainly of thrushes, explained part of the annual variation in cases of neuroborreliosis, with a one-year time lag. We highlight the important role of the genospecies' pathogenicity and the host associations for understanding the epidemiology of disseminated Lyme borreliosis.

MicroRNA let-7 regulates the expression of ecdysteroid receptor (ECR) in Hyalomma asiaticum (Acari: Ixodidae) ticks

BackgroundTicks are blood-sucking arthropods that can transmit diseases to humans and animals. These arthropods are the second most important vectors of pathogens. MicroRNAs are a class of conserved small noncoding RNAs that play regulatory roles in gene expression at the post-transcriptional level. Molting is an important biological process in arthropods. Research on the molting process is important for understanding tick physiology and control.MethodsDual-luciferase reporter assays were used to assess the role of miRNA let-7 in ecdysteroid receptor (ECR) biology. The expression levels of ECR and let-7 were measured by real-time qPCR before and after tick molting. To explore the function of let-7 and ECR, we performed overexpression and knocking down of let-7 and RNAi of ECR in tick nymphs. The biological function of let-7 in molting was explored by injecting nymphs, ten days after engorgement, with let-7 agomir for overexpression and let-7 antagomir for knocking down. The rate of molting was then determined. ECR dsRNA was injected into ticks to evaluate the function of ECR by gene silencing. The expression of ECR and let-7 was measured using RT-qPCR. All data were analyzed using GraphPad Prism v.6.ResultsThe results of the luciferase assay using a eukaryotic expression system revealed that ECR was a natural target of let-7. Let-7 overexpressed by agomir affected the rate of molting (P < 0.01) and the period of molting (P < 0.01). Let-7 antagomir for knockdown affected the period of molting (P < 0.01), but there was no effect on the rate of molting (P = 0.27). ECR dsRNA gene silencing significantly affected the rate of molting (P < 0.05).ConclusionsThis study demonstrated that let-7 can regulate the expression of ECR and that let-7 can affect molting in ticks. Our results help to understand the regulation of let-7 by 20-hydroxyecdysone (20E) and will provide a reference for functional analysis studies of microRNAs in ticks.

Vector Competence for West Nile Virus and St. Louis Encephalitis Virus (Flavivirus) of Three Tick Species of the Genus Amblyomma (Acari: Ixodidae).

Many species of Amblyomma ticks are commonly found infesting wild birds in South America, where birds are important hosts for several arboviruses, such as West Nile virus (WNV) and St. Louis encephalitis virus (SLEV). In this study, WNV and SLEV transmission experiments were performed to evaluate the vector competence of three South American tick species: Amblyomma ovale, Amblyomma tigrinum, and Amblyomma tonelliae. Larval and nymphal ticks of each species were allowed to feed on chicks needle inoculated with WNV or SLEV. All three Amblyomma species acquired either WNV or SLEV through larval feeding, with infection rates varying from 3.1% to 100% for WNV and from 0% to 35.7% for SLEV in engorged larvae. Transstadial perpetuation of the viruses was demonstrated in the molted nymphs, with WNV infection rates varying from 0% to 33.7% and SLEV infection rates from 13.6% to 23.8%. Although nymphal ticks also acquired either virus through feeding, transstadial perpetuation to adult ticks was lower, with virus detection in only 3.2% of A. tigrinum and 11.5% of A. tonelliae unfed adult ticks. On the other hand, vector competence for nymphs (exposed to WNV or SLEV through larval feeding) and adult ticks (exposed to WNV or SLEV through larval or nymphal feeding) was null in all cases. Although our results indicate transstadial perpetuation of WNV or SLEV in the three tick species, the ticks were not competent to transmit these agents to susceptible hosts. The role of these ixodid tick species in the epidemiology of WNV and SLEV might be insignificant, even though at least A. ovale and A. tigrinum are frequent bird ticks in Latin America, so the virus could survive winter in the fed larvae. However, future studies are required to determine the implications that this could have, as well as analyze the vector competence of other common bird tick species in South America.