Inkoo and Sindbis viruses in blood sucking insects, and a serological study for Inkoo virus in semi-domesticated Eurasian tundra reindeer in Norway.

BACKGROUND: Mosquito-borne viruses pose a serious threat to humans worldwide. There has been an upsurge in the number of mosquito-borne viruses in Europe, mostly belonging to the families Togaviridae, genus Alphavirus (Sindbis, Chikungunya), Flaviviridae (West Nile, Usutu, Dengue), and Peribunyaviridae, genus Orthobunyavirus, California serogroup (Inkoo, Batai, Tahyna). The principal focus of this study was Inkoo (INKV) and Sindbis (SINV) virus circulating in Norway, Sweden, Finland, and some parts of Russia. These viruses are associated with morbidity in humans. However, there is a knowledge gap regarding reservoirs and transmission. Therefore, we aimed to determine the prevalence of INKV and SINV in blood sucking insects and seroprevalence for INKV in semi-domesticated Eurasian tundra reindeer (Rangifer tarandus tarandus) in Norway. MATERIALS AND METHODS: In total, 213 pools containing about 25 blood sucking insects (BSI) each and 480 reindeer sera were collected in eight Norwegian reindeer summer pasture districts during 2013-2015. The pools were analysed by RT-PCR to detect INKV and by RT-real-time PCR for SINV. Reindeer sera were analysed for INKV-specific IgG by an Indirect Immunofluorescence Assay (n = 480, IIFA) and a Plaque Reduction Neutralization Test (n = 60, PRNT). RESULTS: Aedes spp. were the most dominant species among the collected BSI. Two of the pools were positive for INKV-RNA by RT-PCR and were confirmed by pyrosequencing. The overall estimated pool prevalence (EPP) of INKV in Norway was 0.04%. None of the analysed pools were positive for SINV. Overall IgG seroprevalence in reindeer was 62% positive for INKV by IIFA. Of the 60 reindeer sera- analysed by PRNT for INKV, 80% were confirmed positive, and there was no cross-reactivity with the closely related Tahyna virus (TAHV) and Snowshoe hare virus (SSHV). CONCLUSION: The occurrence and prevalence of INKV in BSI and the high seroprevalence against the virus among semi-domesticated reindeer in Norway indicate that further studies are required for monitoring this virus. SINV was not detected in the BSI in this study, however, human cases of SINV infection are yearly reported from other regions such as Rjukan in south-central Norway. It is therefore essential to monitor both viruses in the human population. Our findings are important to raise awareness regarding the geographical distribution of these mosquito-borne viruses in Northern Europe.

Correction to: Detection of Candidatus Neoehrlichia mikurensis in Norway up to the northern limit of Ixodes ricinus distribution using a novel real time PCR test targeting the groEL gene.

After publication of our article [1] it came to our notice that the source of the sequence for the control plasmid, pNeo (Materials and methods: Controls) was incorrectly stated as AB094461. The correct accession number is AB074461. The authors apologize for any confusion this may have caused.

Detection of Candidatus Neoehrlichia mikurensis in Norway up to the northern limit of Ixodes ricinus distribution using a novel real time PCR test targeting the groEL gene.

BACKGROUND: Candidatus Neoehrlichia mikurensis is an emerging tick-borne pathogen. It is widely distributed in Ixodes ricinus ticks in Europe, but knowledge of its distribution in Norway, where I. ricinus reaches its northern limit, is limited. In this study we have developed a real time PCR test for Ca. N. mikurensis and used it to investigate the distribution of Ca. N. mikurensis in Norway. RESULTS: Real time PCR targeting the groEL gene was developed and shown to be highly sensitive. It was used to detect Ca. N. mikurensis in 1651 I. ricinus nymphs and adults collected from twelve locations in Norway, from the eastern Oslo Fjord in the south to near the Arctic Circle in the north. The overall prevalence was 6.5% and varied locally between 0 and 16%. Prevalence in adults and nymphs was similar, suggesting that ticks acquire Ca. N. mikurensis predominantly during their first blood meal. In addition, 123 larvae were investigated; Ca. N. mikurensis was not found in larvae, suggesting that transovarial transmission is rare or absent. Sequence analysis suggests that a single variant dominates in Norway. CONCLUSIONS: Ca. N. mikurensis is widespread and common in ticks in Norway and reaches up to their northern limit near the Arctic Circle. Ticks appear to acquire Ca. N. mikurensis during their first blood meal. No evidence for transovarial transmission was found.

Predicting and mapping human risk of exposure to Ixodes ricinus nymphs using climatic and environmental data, Denmark, Norway and Sweden, 2016.

BackgroundTick-borne diseases have become increasingly common in recent decades and present a health problem in many parts of Europe. Control and prevention of these diseases require a better understanding of vector distribution.AimOur aim was to create a model able to predict the distribution of Ixodes ricinus nymphs in southern Scandinavia and to assess how this relates to risk of human exposure.MethodsWe measured the presence of I. ricinus tick nymphs at 159 stratified random lowland forest and meadow sites in Denmark, Norway and Sweden by dragging 400 m transects from August to September 2016, representing a total distance of 63.6 km. Using climate and remote sensing environmental data and boosted regression tree modelling, we predicted the overall spatial distribution of I. ricinus nymphs in Scandinavia. To assess the potential public health impact, we combined the predicted tick distribution with human density maps to determine the proportion of people at risk.ResultsOur model predicted the spatial distribution of I. ricinus nymphs with a sensitivity of 91% and a specificity of 60%. Temperature was one of the main drivers in the model followed by vegetation cover. Nymphs were restricted to only 17.5% of the modelled area but, respectively, 73.5%, 67.1% and 78.8% of the human populations lived within 5 km of these areas in Denmark, Norway and Sweden.ConclusionThe model suggests that increasing temperatures in the future may expand tick distribution geographically in northern Europe, but this may only affect a small additional proportion of the human population.

Head lice predictors and infestation dynamics among primary school children in Norway.

BACKGROUND: Health providers need to know which measures to take and children to prioritize in order to decrease costs associated with head lice infestations. OBJECTIVE: Our aim was to determine the most important predictors for head lice and identify the major drivers of an infestation outbreak in a low-prevalence area. METHODS: The study was based on three datasets of head lice prevalence (retrospective, point prevalence and prospective approach) from primary school children (ages 6-12) at 12 schools in Oslo, Norway. The tested predictors were siblings with lice, individual and household characteristics as well as class and school affiliation. Self-reported monthly incidences (prospective approach) of head lice were used to evaluate infestation dynamics. RESULTS: Infested siblings strongly increased the odds of head lice infestation of school children (odds ratio 36, 26 and 7 in the three datasets) whereas having short hair halved the odds. Household characteristics were of minor importance, and class affiliation proved more important than school affiliation. Having head lice in one school term increased the odds of an infestation in the next, but this effect diminished over time. About 97% of all self-reported infestations were noted in two consecutive months or less. CONCLUSIONS: With the exception of hair length, we have found that individual and household characteristics are of minor importance to predict head lice infestations in a low-prevalence country and that unnoticed transmissions in school classes and families are likely to be the major driver upon outbreaks.

Detection of specific IgG antibodies in blood donors and tick-borne encephalitis virus in ticks within a non-endemic area in southeast Norway.

BACKGROUND: Tick-borne encephalitis (TBE) is an emerging tick-borne disease in Europe. In Norway, the first TBE case occurred in 1997, and since then 1-14 cases have been detected annually along the southern coast. No TBE cases have yet been notified from the eastern coastal area. This study was conducted to assess the need for diagnostic tests and vaccine recommendation for this part of Norway. METHODS: Four hundred and sixty-one blood donors living in the county of Østfold were enrolled. After informed consent was obtained, the participants submitted a blood sample and filled out a questionnaire regarding tick bites, outdoor activities, and Flavivirus vaccines and diseases. Ixodes ricinus ticks were collected from the immediate vicinity and were examined in pools of 10 for TBE virus. RESULTS: Eight human samples were TBE virus IgG-positive by ELISA and 5 of these samples were confirmed positive by neutralization test. Excluding the 2 samples from participants who had reported previous TBE vaccination, this shows a seroprevalence among blood donors of 0.65%. The existence of TBEV in the region was verified in nymphs of Ixodes ricinus by a prevalence of 0.14%. CONCLUSIONS: The seroprevalence of TBE virus IgG and the TBE virus detected in ticks, indicate that TBE cases could occasionally occur in the area. The results should be made available to health care personnel to raise awareness for preventative measures.

Socioeconomic status, family background and other key factors influence the management of head lice in Norway.

How head lice infestations are managed by households is an important but generally neglected issue in head lice research. In the present study, we investigate actions taken against head lice by Norwegian households in association with socioeconomic status, family background, school-related variables and other key factors. Repeat questionnaires distributed to caretakers of the same elementary school children during a 2-year period enabled us to study both previous head lice management and any changes in this management through time. Households from 12 schools spanning the main socioeconomic variation found in Norway participated in the study. All students with active head lice infestation were treated in the four investigated periods. Most caretakers used a thorough head lice checking technique and informed others of own infestation. Checking frequency was low as most children were inspected less than monthly. The best determinant of increased checking frequency and thoroughness was personal experience with head lice. The increased awareness, however, seemed to be somewhat short-lived, as there was a decrease in checking frequency and thoroughness within 1 year after infestation. Personal experience with head lice also increased general knowledge related to the parasite. Parents born in developing countries checked their children for head lice more frequently, although less thoroughly, informed fewer contacts when infested, used pediculicides preventively more often and knew less about head lice than parents born in developed countries. Households with highly educated mothers had a lower checking frequency, but their knowledge and willingness to inform others was high. Single parents were more concerned about economic costs and kept children home from school longer while infested than other parents. As head lice management varied among socioeconomic groups and with parental background, differentiated advice should be considered in the control of head lice. The biannual focus on head lice during the 2 years of investigation increased checking thoroughness, while checking frequency remained unchanged. Based on the results, we suggest new head lice management guidelines for health authorities.

Serological screening for tick-borne encephalitis virus in eight Norwegian herds of semi-domesticated reindeer (Rangifer tarandus tarandus).

Tick-borne encephalitis virus (TBEV) is found in Ixodes ricinus ticks throughout the area where viable tick populations exist. In Norway, TBEV is found in I. ricinus from the south coast until Brønnøy municipality in Nordland County and the range of the vector is expanding due to changes in climate, vegetation, host animals and environmental conditions. TBEV might thus have the potential to establish in new areas when I. ricinus expand its geographical distribution. At present, there is little knowledge on the status of the virus in high-altitude areas of inland regions in Norway. It has previously been indicated that reindeer may be an important sentinel species and indicator of the spread of ticks and TBEV in high-altitude regions. In this study, 408 semi-domesticated Eurasian tundra reindeer (Rangifer tarandus tarandus) from eight herds, from Tana in Troms and Finnmark County in northern Norway to Filefjell in Innlandet and Viken Counties in southern Norway, were screened for TBEV antibodies using a commercial enzyme-linked immunosorbent assay (ELISA). We found 16 TBEV reactive reindeer samples by ELISA; however, these results could not be confirmed by the serum neutralization test (SNT). This could indicate that a flavivirusand not necessarily TBEV, may be circulating among Norwegian semi-domesticated reindeer. The results also indicate that TBEV was not enzootic in Norwegian semi-domesticated reindeer in 2013-2015. This knowledge is important as an information base for future TBEV and flavivirus surveillance in Norway.

Prevalence of tick-borne encephalitis virus in questing Ixodes ricinus nymphs in southern Scandinavia and the possible influence of meteorological factors.

Ixodes ricinus ticks are Scandinavia's main vector for tick-borne encephalitis virus (TBEV), which infects many people annually. The aims of the present study were (i) to obtain information on the TBEV prevalence in host-seeking I. ricinus collected within the Øresund-Kattegat-Skagerrak (ØKS) region, which lies in southern Norway, southern Sweden and Denmark; (ii) to analyse whether there are potential spatial patterns in the TBEV prevalence; and (iii) to understand the relationship between TBEV prevalence and meteorological factors in southern Scandinavia. Tick nymphs were collected in 2016, in southern Scandinavia, and screened for TBEV, using pools of 10 nymphs, with RT real-time PCR, and positive samples were confirmed with pyrosequencing. Spatial autocorrelation and cluster analysis was performed with Global Moran's I and SatScan to test for spatial patterns and potential local clusters of the TBEV pool prevalence at each of the 50 sites. A climatic analysis was made to correlate parameters such as minimum, mean and maximum temperature, relative humidity and saturation deficit with TBEV pool prevalence. The climatic data were acquired from the nearest meteorological stations for 2015 and 2016. This study confirms the presence of TBEV in 12 out of 30 locations in Denmark, where six were from Jutland, three from Zealand and two from Bornholm and Falster counties. In total, five out of nine sites were positive from southern Sweden. TBEV prevalence of 0.7%, 0.5% and 0.5%, in nymphs, was found at three sites along the Oslofjord (two sites) and northern Skåne region (one site), indicating a potential concern for public health. We report an overall estimated TBEV prevalence of 0.1% in questing I. ricinus nymphs in southern Scandinavia with a region-specific prevalence of 0.1% in Denmark, 0.2% in southern Sweden and 0.1% in southeastern Norway. No evidence of a spatial pattern or local clusters was found in the study region. We found a strong correlation between TBEV prevalence in ticks and relative humidity in Sweden and Norway, which might suggest that humidity has a role in maintaining TBEV prevalence in ticks. TBEV is an emerging tick-borne pathogen in southern Scandinavia, and we recommend further studies to understand the TBEV transmission potential with changing climate in Scandinavia.

Suspected rodenticide exposures in humans and domestic animals: Data from inquiries to the Norwegian Poison Information Centre, 2005-2020.

Rodent control is necessary to prevent damage and spread of disease, and the most common pesticides used for urban and rural rodent control are anticoagulant rodenticides. The aim of this present study was to present data on suspected exposure to rodenticides in humans and domestic animals in Norway based on inquiries to the Norwegian Poison Information Centre in the 16-year period from 2005 through 2020. A total of 4235 inquiries regarding suspected exposures to rodenticides were registered in the study period. Of these, 1486 inquiries involved humans and 2749 animals. Second generation anticoagulants were involved in 68% of human exposures and 79% of animal exposures. Dogs were the most frequent species involved in the animal exposures with 93% of the inquiries, while cats were second most frequent involved. Around 50% of the human inquiries concerned children at the age of 0-4 years. Only 2% of the cases were in the age group 10-19 years, while adults comprised 35% of the inquiries. Acute poisonings accounted for almost 100% of the inquiries among both humans and animals. The exposure was accidental in 99% of the animal exposures and in 85% of the human exposures. In humans, only 14 inquiries were regarding occupational related accidents. Misdeed or self-inflicted injury accounted for 15% of the human inquiries and were the cause of 79% of the severe poisonings. Severe poisoning was only assessed in 1% of the cases involving children under 5 years. In contrast, 17% of the inquiries concerning adults (≥20 years) were assessed as severe. Subsequently, to prevent human and animal rodenticide exposure, we urge the use of non-chemical methods such as sanitation, rodent proofing (a form of construction which will impede or prevent rodents access to or from a given space or building) and mechanical traps. Restricting the use of rodenticides to professional pest controllers (or other persons with authorisation), reinforcing high quality education of these persons, and securing compliance of the best codes of practice could be advocated to reduce accidental exposure to rodenticides in humans and animals.

Head lice prevalence among households in Norway: importance of spatial variables and individual and household characteristics.

Head lice prevalence varies greatly between and within countries, and more knowledge is needed to approach causes of this variation. In the present study, we investigated head lice prevalence among elementary school students and their households in relation to individual and household characteristics as well as spatial variables. The investigation included households from 5 geographically separated municipalities. Present infestations among household members as well as previous infestations in the household were reported in a questionnaire. In elementary school students prevalence was low (1·63%), but more than one-third of the households (36·43%) had previously experienced pediculosis. Prevalence was higher in elementary school students than in other household members, and highest in third-grade children. Prevalence was also influenced by the school attended, which suggested that interactions between children in the same school are important for head lice transmission. Previous occurrence of head lice in homes also increased the risk of present infestation. Prevalence of previous infestations was higher in households with more children and in more densely populated municipalities, indicating that the density of hosts or groups of hosts influences transmission rates. These results demonstrate that information of hosts' spatial distribution as well as household and individual characteristics is needed to better understand head lice population dynamics.

Distribution of Neoehrlichia mikurensis in Ixodes ricinus ticks along the coast of Norway: The western seaboard is a low-prevalence region.

Neoehrlichia mikurensis is a tick-borne pathogen widespread among ticks and rodents in Europe and Asia. A previous study on Ixodes ricinus ticks in Norway suggested that N. mikurensis was scarce or absent on the south-west coast of Norway, but abundant elsewhere. The aim of this study was to further investigate the prevalence and distribution of N. mikurensis along the western seaboard of Norway in comparison with more eastern and northern areas. The second aim of the study was to examine seasonal variation of the bacterium in one specific location in the south-eastern part of Norway. Questing I. ricinus were collected from 13 locations along the coast of Norway, from Brønnøysund in Nordland County to Spjaerøy in Østfold County. In total, 11,113 nymphs in 1,113 pools and 718 individual adult ticks were analysed for N. mikurensis by real-time PCR. The mean prevalence of N. mikurensis in adult ticks was 7.9% while the estimated pooled prevalence in nymphs was 3.5%. The prevalence ranged from 0% to 25.5%, with the highest prevalence in the southernmost and the northernmost locations. The pathogen was absent, or present only at low prevalence (<5%), at eight locations, all located in the west, from 58.9°N to 64.9°N. The prevalence of N. mikurensis was significantly different between counties (p < .0001). No significant seasonal variation of N. mikurensis prevalence was observed in the period May to October 2015. Our results confirm earlier findings of a low prevalence of N. mikurensis in the western seaboard of Norway.

Author Correction: Spatial data of Ixodes ricinus instar abundance and nymph pathogen prevalence, Scandinavia, 2016-2017.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Spatial data of Ixodes ricinus instar abundance and nymph pathogen prevalence, Scandinavia, 2016-2017.

Ticks carry pathogens that can cause disease in both animals and humans, and there is a need to monitor the distribution and abundance of ticks and the pathogens they carry to pinpoint potential high risk areas for tick-borne disease transmission. In a joint Scandinavian study, we measured Ixodes ricinus instar abundance at 159 sites in southern Scandinavia in August-September, 2016, and collected 29,440 tick nymphs at 50 of these sites. We additionally measured abundance at 30 sites in August-September, 2017. We tested the 29,440 tick nymphs in pools of 10 in a Fluidigm real-time PCR chip to screen for 17 different tick-associated pathogens, 2 pathogen groups and 3 tick species. We present data on the geolocation, habitat type and instar abundance of the surveyed sites, as well as presence/absence of each pathogen in all analysed pools from the 50 collection sites and individual prevalence for each site. These data can be used alone or in combination with other data for predictive modelling and mapping of high-risk areas.

Spatial patterns of pathogen prevalence in questing Ixodes ricinus nymphs in southern Scandinavia, 2016.

Tick-borne pathogens cause diseases in animals and humans, and tick-borne disease incidence is increasing in many parts of the world. There is a need to assess the distribution of tick-borne pathogens and identify potential risk areas. We collected 29,440 tick nymphs from 50 sites in Scandinavia from August to September, 2016. We tested ticks in a real-time PCR chip, screening for 19 vector-associated pathogens. We analysed spatial patterns, mapped the prevalence of each pathogen and used machine learning algorithms and environmental variables to develop predictive prevalence models. All 50 sites had a pool prevalence of at least 33% for one or more pathogens, the most prevalent being Borrelia afzelii, B. garinii, Rickettsia helvetica, Anaplasma phagocytophilum, and Neoehrlichia mikurensis. There were large differences in pathogen prevalence between sites, but we identified only limited geographical clustering. The prevalence models performed poorly, with only models for R. helvetica and N. mikurensis having moderate predictive power (normalized RMSE from 0.74-0.75, R2 from 0.43-0.48). The poor performance of the majority of our prevalence models suggest that the used environmental and climatic variables alone do not explain pathogen prevalence patterns in Scandinavia, although previously the same variables successfully predicted spatial patterns of ticks in the same area.

Geographical distribution and prevalence of tick-borne encephalitis virus in questing Ixodes ricinus ticks and phylogeographic structure of the Ixodes ricinus vector in Norway.

The tick-borne encephalitis virus (TBEV), a zoonotic flaviviral infection, is endemic in large parts of Norway and Eurasia. Humans are mainly infected with TBEV via bites from infected ticks. In Norway, the main geographical distribution of ticks is along the Norwegian coastline from southeast (~59°N) and up to the southern parts of Nordland County (~65°N). In this study, we collected ticks by flagging along the coast from Østfold County to Nordland County. By whole-genome sequencing of the mitochondrial genome of Ixodes ricinus, the phylogenetic tree suggests that there is limited phylogeographic structure both in Norway and in Europe. The overall TBEV prevalence is 0.3% for nymphs and 4.3% for adults. The highest estimated TBEV prevalence in adult ticks was detected in Rogaland and Vestfold County, while for nymphs it is highest in Vestfold, Vest-Agder and Rogaland. The present work is one of the largest studies on distribution and prevalence of TBEV in ticks in Scandinavia, showing that the virus is wider distributed in Norway than previously anticipated.

Tick-borne encephalitis virus in cows and unpasteurized cow milk from Norway.

Tick-borne encephalitis virus (TBEV) is recognized as the most important zoonotic tick-transmitted virus in Europe. TBEV is mainly transmitted to humans through bites from TBEV-infected ticks (Ixodes ricinus and Ixodes persulcatus). However, alimentary infection after consumption of unpasteurized milk and cheese from domestic ruminants has been reported. There is little information about TBEV in ruminants in Norway. The objectives of this study were to analyse unpasteurized cow milk for TBEV RNA and to study the presence of IgG antibodies to TBEV in the same animals. A total of 112 milk and blood samples were collected from cows from five different farms spread from southern to northern Norway. The milk samples were analysed by an in-house reverse transcription (RT) real-time polymerase chain reaction and confirmed by pyrosequencing. Serum samples were screened by a commercial enzyme-linked immunosorbent assay and verified by a TBEV-specific serum neutralization test. We found TBEV RNA in unpasteurized milk collected from farms in the municipalities of Mandal, Skedsmo and Brønnøy in 5.4% of the tested animals. Specific antibodies to TBEV were only detected in Arendal, where 88.2% of the tested animals were positive. Further studies on milk containing TBEV RNA should be performed to conclude if TBEV found in unpasteurized milk in Norway is infectious, which could be of great importance in a One Health perspective.

Predicting the spatial abundance of Ixodes ricinus ticks in southern Scandinavia using environmental and climatic data.

Recently, focus on tick-borne diseases has increased as ticks and their pathogens have become widespread and represent a health problem in Europe. Understanding the epidemiology of tick-borne infections requires the ability to predict and map tick abundance. We measured Ixodes ricinus abundance at 159 sites in southern Scandinavia from August-September, 2016. We used field data and environmental variables to develop predictive abundance models using machine learning algorithms, and also tested these models on 2017 data. Larva and nymph abundance models had relatively high predictive power (normalized RMSE from 0.65-0.69, R2 from 0.52-0.58) whereas adult tick models performed poorly (normalized RMSE from 0.94-0.96, R2 from 0.04-0.10). Testing the models on 2017 data produced good results with normalized RMSE values from 0.59-1.13 and R2 from 0.18-0.69. The resulting 2016 maps corresponded well with known tick abundance and distribution in Scandinavia. The models were highly influenced by temperature and vegetation, indicating that climate may be an important driver of I. ricinus distribution and abundance in Scandinavia. Despite varying results, the models predicted abundance in 2017 with high accuracy. The models are a first step towards environmentally driven tick abundance models that can assist in determining risk areas and interpreting human incidence data.

A large-scale screening for the taiga tick, Ixodes persulcatus, and the meadow tick, Dermacentor reticulatus, in southern Scandinavia, 2016.

The taiga tick, Ixodes persulcatus, has previously been limited to eastern Europe and northern Asia, but recently its range has expanded to Finland and northern Sweden. The species is of medical importance, as it, along with a string of other pathogens, may carry the Siberian and Far Eastern subtypes of tick-borne encephalitis virus. These subtypes appear to cause more severe disease, with higher fatality rates than the central European subtype. Until recently, the meadow tick, Dermacentor reticulatus, has been absent from Scandinavia, but has now been detected in Denmark, Norway and Sweden. Dermacentor reticulatus carries, along with other pathogens, Babesia canis and Rickettsia raoultii. Babesia canis causes severe and often fatal canine babesiosis, and R. raoultii may cause disease in humans. We collected 600 tick nymphs from each of 50 randomly selected sites in Denmark, southern Norway and south-eastern Sweden in August-September 2016. We tested pools of 10 nymphs in a Fluidigm real time PCR chip to screen for I. persulcatus and D. reticulatus, as well as tick-borne pathogens. Of all the 30,000 nymphs tested, none were I. persulcatus or D. reticulatus. Our results suggest that I. persulcatus is still limited to the northern parts of Sweden, and have not expanded into southern parts of Scandinavia. According to literature reports and supported by our screening results, D. reticulatus may yet only be an occasional guest in Scandinavia without established populations.

Tick-borne encephalitis virus, Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Anaplasma phagocytophilum and Candidatus Neoehrlichia mikurensis in Ixodes ricinus ticks collected from recreational islands in southern Norway.

The aim of this study was to determine the occurrence of tick-borne pathogens of medical importance in questing ticks collected from five recreationally used islands along the Norwegian coastline. Furthermore, since coinfection may affect the disease severity, this study aimed to determine the extent of coinfection in individual ticks or co-localization of tick-borne pathogens. In all, 4158 questing Ixodes ricinus ticks were analyzed. For detection of tick-borne encephalitis virus (TBEV), nymphs (3690) were analyzed in pools of ten. To detect Borrelia burgdorferi sensu lato, B. miyamotoi, Anaplasma phagocytophilum and Candidatus Neoehrlichia mikurensis, 468 nymphs were analyzed individually. A total of five nymph pools was infected with TBEV, giving an overall prevalence of 0.14%. In the individually analyzed ticks, B. burgdorferi s. l. (15.6%), Candidatus N. mikurensis (11%), A. phagocytophilum (1.4%) and B. miyamotoi (0.9%) were detected. Coinfection was found in 3.3% of the ticks, and the only dual infection observed was with B. afzelii and Candidatus N. mikurensis. This association was significantly higher than what would occur by random chance.