Phagocytophilum Prevalence Research Articles

Anaplasma phagocytophilum in Ticks Blood-Feeding on Migratory Birds in Sweden.

Migratory birds play a dual role as potential reservoirs of tick-borne pathogens, and potential dispersers of pathogen-containing ticks during their migratory journeys. Ixodes ricinus, a prevalent tick species in Northern and Western Europe, serves as a primary vector for Anaplasma phagocytophilum-a bacterium with implications for human and animal health. There is limited information available regarding A. phagocytophilum in birds. Our investigation focused on A. phagocytophilum prevalence in ticks collected from migratory birds in southeastern Sweden. The identification of ticks involved both molecular analyses for species determination and morphological classification to ascertain the developmental stage. The presence of A. phagocytophilum was determined using real-time PCR. Of the 1115 ticks analyzed from 4601 birds, 0.9% (n = 10), including I. ricinus and Ixodes frontalis, tested positive for A. phagocytophilum. Notably, common blackbirds (Turdus merula) yielded the highest number of A. phagocytophilum-infected ticks. The findings suggest that A. phagocytophilum is present in a small proportion of ticks infesting migratory birds in southeastern Sweden. Consequently, the role of birds as hosts for ticks infected with A. phagocytophilum appears to be low, suggesting that birds seem to play a minor indirect role in the geographic dispersal of A. phagocytophilum.

Frequency of Anaplasma phagocytophilum, Borrelia spp., and coinfections in Ixodes ricinus ticks collected from dogs and cats in Germany

BackgroundChanging geographical and seasonal activity patterns of ticks may increase the risk of tick infestation and tick-borne pathogen (TBP) transmission for both humans and animals.MethodsTo estimate TBP exposure of dogs and cats, 3000 female I. ricinus from these hosts were investigated for Anaplasma phagocytophilum and Borrelia species.ResultsqPCR inhibition, which was observed for ticks of all engorgement stages but not questing ticks, was eliminated at a template volume of 2 µl. In ticks from dogs, A. phagocytophilum and Borrelia spp. prevalence amounted to 19.0% (285/1500) and 28.5% (427/1500), respectively, while ticks from cats showed significantly higher values of 30.9% (464/1500) and 55.1% (827/1500). Accordingly, the coinfection rate with both A. phagocytophilum and Borrelia spp. was significantly higher in ticks from cats (17.5%, 262/1500) than dogs (6.9%, 104/1500). Borrelia prevalence significantly decreased with increasing engorgement duration in ticks from both host species, whereas A. phagocytophilum prevalence decreased only in ticks from dogs. While A. phagocytophilum copy numbers in positive ticks did not change significantly over the time of engorgement, those of Borrelia decreased initially in dog ticks. In ticks from cats, copy numbers of neither A. phagocytophilum nor Borrelia spp. were affected by engorgement. Borrelia species differentiation was successful in 29.1% (365/1254) of qPCR-positive ticks. The most frequently detected species in ticks from dogs were B. afzelii (39.3% of successfully differentiated infections; 70/178), B. miyamotoi (16.3%; 29/178), and B. valaisiana (15.7%; 28/178), while B. afzelii (40.1%; 91/227), B. spielmanii (21.6%; 49/227), and B. miyamotoi (14.1%; 32/227) occurred most frequently in ticks from cats.ConclusionsThe differences in pathogen prevalence and Borrelia species distribution between ticks collected from dogs and cats may result from differences in habitat overlap with TBP reservoir hosts. The declining prevalence of A. phagocytophilum with increasing engorgement duration, without a decrease in copy numbers, could indicate transmission to dogs over the time of attachment. The fact that this was not observed in ticks from cats may indicate less efficient transmission. In conclusion, the high prevalence of A. phagocytophilum and Borrelia spp. in ticks collected from dogs and cats underlines the need for effective acaricide tick control to protect both animals and humans from associated health risks.Graphical

A 15-year monitoring of Rickettsiales (Anaplasma phagocytophilum and Rickettsia spp.) in questing ticks in the city of Hanover, Germany

Rickettsiales (Anaplasma phagocytophilum and Rickettsia spp.) are regarded as potentially emerging tick-borne pathogens and may change in abundance in response to global climate change. However, continuous monitoring on their prevalence in questing ticks is only available for the northern German city of Hanover. In the presented follow-up of this long-term study, 2100 questing ticks of the Ixodes ricinus/Ixodes inopinatus-complex collected from April to October 2020 at ten different recreation sites in Hanover were individually analysed for Rickettsia and A. phagocytophilum infection by quantitative real-time PCR. Together with previous results from years 2005, 2010 and 2015, the current study allows to assess potential changes in tick infection rates with Rickettsiales over a 15-year monitoring period. In 2020, 3.0% (63/2100) of ticks were infected with A. phagocytophilum, 36.0% (756/2100) with Rickettsia spp. and 1.2% (26/2100) with both pathogens. Regarding the different developmental tick stages, nymphs showed a significantly lower A. phagocytophilum prevalence of 0.5% (5/1050) than adult ticks (5.5% [58/1050]) as well as compared to females (5.4% [38/700]) and males (5.7% [20/350]). For Rickettsia spp., nymphs also showed a lower prevalence of 33.2% (349/1050) with a significant difference to adult ticks (38.8% [407/1050]) and female ticks (40.7% [285/700]), while males had a Rickettsia infection rate of 34.9% (122/350). Comparison with previous years indicated a stable A. phagocytophilum prevalence over the 15-year monitoring period. In contrast, fluctuating Rickettsia prevalences were observed, with a peak in 2015 in all developmental stages, but similar infection rates in 2005 and 2020. Therefore, epidemiological changes in response to climate change are not (yet) evident. Nevertheless, the long-term monitoring study will be continued in the future, as climatic impacts on tick and reservoir host populations may have a delayed effect on pathogen prevalence and, consequently, transmission to humans and domestic animals.

Prevalence of Anaplasma phagocytophilum in questing Ixodes ricinus nymphs across twenty recreational areas in England and Wales

Human granulocytic anaplasmosis and tick-borne fever, affecting livestock, are diseases caused by an infection with the bacterium Anaplasma phagocytophilum. Its transmission dynamics between vertebrate hosts and ticks remain largely unknown and the potential impact on public health in the United Kingdom is unclear. This study aimed to assess the distribution and estimate the prevalence of A. phagocytophilum in questing Ixodes ricinus at recreational locations across England and Wales over six years. An additional objective was to investigate possible associations between prevalence, habitat and presence of ruminant hosts. Ixodes ricinus ticks were collected each spring at 20 recreational locations across England and Wales between 2014 and 2019. Nymphs were tested for infection with A. phagocytophilum by detection of bacterial genome in DNA extracts, targeting the msp2 gene locus. Positive samples were further investigated for the presence of different ecotypes based on the GroEL region. Of 3,919 nymphs tested, the mean infection prevalence was 3.6% [95%CI: 3.1-4.3] and ranged from 0 to 20.4%. Northern England had a higher overall prevalence (4.7% [95%CI: 3.4-6.4]) compared to Southern England (1.8% [95%CI: 1.3-2.5]) and the presence of sheep was associated with higher A. phagocytophilum prevalence (8.4% [95%CI: 6.9-10.1] vs 1.2% [95%CI: 0.8-1.7] when absent). There was also a negative correlation with the prevalence of Borrelia burgdorferi s.l. (causing Lyme borreliosis). When investigating the diversity of A. phagocytophilum, ecotype I accounted for 86.8% of samples and ecotype II for 13.2%. Our study presents an overview of A. phagocytophilum prevalence in questing I. ricinus in recreational areas across England and Wales and discusses the potential public and veterinary health relevance.

Relationships between landscape structure and the prevalence of two tick-borne infectious agents, Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato, in small mammal communities

By modifying ecosystems, land cover changes influence the emergence, the spread and the incidence of vector-borne diseases. This study aimed at identifying associations between landscape structure and the prevalence of two tick-borne infectious agents, Anaplasma phagocytophilum and Borrelia burgdorferi s.l., in small mammal communities. Small mammals were sampled in 24 sites along a gradient of woodland fragmentation and hedgerow network density, and screened for infectious agents with real-time PCR techniques. For each site, structural variables (composition and configuration) of the surrounding landscape at various scales (0–500 m) and variables of wooded habitats connectivity based on graph theory and least cost path distances for the two dominant species, bank voles (Myodes glareolus) and wood mice (Apodemus sylvaticus), were computed. The A. phagocytophilum prevalence increased with wooded habitats cover (0–500 m), likely through host population size, and increased slightly with bank vole abundance, which has a higher reservoir competence than wood mouse. The B. burgdorferi s.l. prevalence increased with wooded ecotones only at local scales (50–100 m). Wooded habitats connectivity measures did not improve models built with simple land cover variables. A more marked spatial pattern was observed for the prevalence of A. phagocytophilum than B.burgdorferi s.l. This study highlights the interest of considering together the ecology of infectious agents (e.g. host specificity) and the host species community ecology to better understand the influence of the landscape structure on the spatial distribution of vector-borne infectious agents.

Prevalence and molecular characterization of Anaplasma phagocytophilum in roe deer (Capreolus capreolus) from Spain

Anaplasma phagocytophilum can infect a wide range of vertebrates; nevertheless, some genetic variants are associated with particular species of tick vectors and animal hosts. It has been suggested that roe deer (Capreolus capreolus) mainly acts as a reservoir of several A. phagocytophilum non-pathogenic variants for other animal species. The aim of this study was to determine the prevalence and identify the genetic variants of A. phagocytophilum in roe deer from Spain in order to assess host-pathogen associations and their pathogenic potential.The spleens of 212 roe deer hunted in Spain were individually collected and analysed by a commercial qPCR kit in order to detect the presence of A. phagocytophilum DNA. Positive samples were further characterized at groESL, 16S rRNA and msp2 partial genes. The possible influence of several intrinsic (age and sex) and extrinsic factors (ecological area) on A. phagocytophilum prevalence was analysed using a logistic regression.Overall, 41.5 % of the samples resulted positive to A. phagocytophilum. The percentage of infected roe deer was significantly higher in the Mediterranean and Oceanic areas than in the Continental and Mountain regions; nevertheless, prevalence was not related to age or sex. Sequence analysis at groESL and 16S rRNA genes allowed the identification of three ecotypes (I to III) and four variants (“Y”, “X”, “W”, “I”), respectively. A high percentage of roe deer from Spain is infected with different variants of A. phagocytophilum; these results have implications for public and animal health since some of these ecotypes and variants have been previously identified in both human and animal clinical cases.

PCR detection of Anaplasma phagocytophilum in stray dogs in Batman, Turkey

Anaplasma phagocytophilum causes high-grade fever in humans, ruminants, horses, cats, and dogs. A. phagocytophilum, which causes granulocytic anaplasmosis in dogs, has a wide geographical distribution in the world, but there are not many studies on the Anaplasma species in dogs in Turkey. A. phagocytophilum, which infects leukocytes in dog, might also be zoonotic. This study aimed to investigate the A. phagocytophilum prevalence in stray dogs in Batman province of Turkey using Polymerase Chain Reaction (PCR) technique. Blood samples were drawn into EDTA tubes from the venae cephalica antebrachii of randomly selected 97 asymptomatic dogs in total. Identification of A. phagocytophilum was carried out using the conventional PCR method that was conducted by using the DNAs obtained from the samples. A. phagocytophilum prevalence was found to be 3.1% in the examined blood samples. In conclusion, infection of dogs in Batman province with A. phagocytophilum has been revealed molecularly for the first time with this study. It is considered that detection of this species, which can cause disease (Human granulocytic anaplasmosis) in humans as well, has contributed epidemiologically to the data on the spread of the disease throughout Turkey. Moreover, investigating the tick species that transmit the disease in the region where the study was conducted is important for developing effective control strategies against the disease.

666. Human Granulocytic Anaplasmosis and Ehrlichiosis Presenting to an Upstate New York Emergency Department

BackgroundHuman granulocytic anaplasmosis (HGA) is an increasingly common tickborne infection in the North-East United States. Statewide incidence of human cases of anaplasmosis in New York (excluding New York City) increased 407% from 2010–2017, with a corresponding increase in Anaplasma phagocytophilum prevalence in ticks. While previous observational studies have described the clinical features of HGA, there has been little documentation of HGA diagnosed in the emergency department (ED) setting.MethodsDemographic and clinical data were extracted from electronic records of cases with a positive polymerase chain reaction (PCR) for HGA or the closely related clinical entity ehrlichiosis from 2016 to 2017. HGA and ehrlichiosis PCR were performed by the Albany Medical Center laboratory on patients treated in the ED. Basic descriptive analyses were performed.ResultsDuring the 2-year period, there were 37 cases of PCR-positive HGA and four cases of PCR-positive ehrlichiosis treated in the ED. Electronic data were available for extraction for 40 of these cases. Mean age was 54 years old (range 7–94 years). Thirty-four (85%) patients presented with fever, and 9 (23%) reported a tick bite or tick exposure. Twenty-two (55%) patients had leukopenia; 32 (80%) thrombocytopenia; 26 (65%) hyponatremia; 24 (60%) elevated transaminases; 38 (95%) at least one of these laboratory abnormality; and 13 (33%) all four laboratory abnormalities. Twenty-four patients (60%) were given the empiric diagnosis of a tickborne illness upon disposition from the ED, with 19 (48%) patients admitted to the hospital.ConclusionTo our knowledge, this study represents the first description of patients diagnosed with HGA (and ehrlichiosis) in the ED setting. Because of the inherent testing bias, further study is needed to establish the true ED prevalence of HGA in highly endemic regions. Notably, only 23% reported either a tick bite or tick exposure, highlighting the need to consider this for diagnosis in patients presenting to the ED with consistent symptoms and laboratory findings in endemic areas. Further study might explore whether there exists a collection of laboratory findings that could accurately identify HGA in ED patients.Disclosures All authors: No reported disclosures.

Seasonal Patterns in the Prevalence and Diversity of Tick-Borne Borrelia burgdorferi Sensu Lato, Anaplasma phagocytophilum and Rickettsia spp. in an Urban Temperate Forest in South Western Slovakia.

In Europe, Ixodes ricinus is the most important vector of tick-borne zoonotic bacteria. It transmits spirochaetes from the Borrelia burgdorferi sensu lato complex, Anaplasma phagocytophilum and Rickettsia spp. Although spatial differences in the prevalence of tick-borne pathogens have been intensively studied, seasonal (within-year) fluctuations in the prevalence of these pathogens within sites are often overlooked. We analyzed the occurrence and seasonal dynamics of Ixodes ricinus in an urban forest in Bratislava, Slovakia. Furthemore, we examined temporal trends in the community structure of B. burgdorferi s.l., A. phagocytophilum and Rickettsia spp. in questing and bird-feeding ticks. The total prevalence for B. burgdorferi s.l. in questing I. ricinus was 6.8%, involving six genospecies with the dominance of bird-associated B. garinii and B. valaisiana. A. phagocytophilum, R. helvetica and R. monacensis occurred in 5.9%, 5.0% and 0.2% of questing ticks, respectively. In total, 12.5% and 4.4% of bird-feeding I. ricinus ticks carried B. burgdorferi s.l. and R. helvetica. The total prevalence of B. burgdorferi s.l. in our study site was two times lower than the mean prevalence for Europe. In contrast, A. phagocytophilum prevalence was significantly higher compared to those in other habitats of Slovakia. Our results imply that tick propagation and the transmission, suppression and seasonal dynamics of tick-borne pathogens at the study site were primarily shaped by abundance and temporal population fluctuations in ruminant and bird hosts.

Altitude-Dependent Prevalence of Canine Granulocytic Anaplasmosis in Romania.

Canine granulocytic anaplasmosis (CGA) is an important tick-borne disease with worldwide distribution. The importance of this disease resides in the ability of Anaplasma phagocytophilum to infect humans and several animal species. The aim of the study was to evaluate the prevalence rate of CGA in different altitudinal areas of Romania. A total of 357 canine blood samples were collected during 2010-2013 from eight counties. To assess the influence of the altitude on A. phagocytophilum prevalence, the samples were collected from four different altitude areas (coastal 0-5 meters; lowland 6-100 meters; hilly areas 200-300 meters; low mountain areas >500 meters). These samples were evaluated for the presence of A. phagocytophilum DNA by amplifying part of the Ankyrin repeat protein (AnkA) gene. A higher prevalence was obtained for coastal compared with remaining areas, suggesting an influence of altitude on the CGA. Moreover, the results suggest an influence of climate and rainfall. In the present research work, we highlight the risk of granulocytic anaplasmosis in Central and Southern Romania, with a greater risk associated to Southern lowland region, especially in coastal areas. The importance of these results resides in the zoonotic potential of the canine A. phagocytophilum strains. In conclusion, the altitude and precipitation level may be risk factors for A. phagocytophilum infection in dogs and other hosts, including humans.

Prevalence of Anaplasma phagocytophilum in small rodents in France

Anaplasma phagocytophilum is an emerging zoonotic tick-borne pathogen affecting a wide range of mammals. Rodents are suspected to be natural reservoirs for this bacterium, but their role in the epidemiologic cycles affecting domestic animals and wild ungulates has not been demonstrated. This study aimed to improve our knowledge on A. phagocytophilum prevalence in Apodemus sylvaticus, A. flavicollis and Myodes glareolus using data collected in 2010 in one area in eastern France and in 2012–2013 in two others areas in western France. Rodents were captured in each site and infection was tested using qualitative real-time PCR assays on either blood or spleen samples. Prevalence showed high variability among sites. The highest prevalence was observed in the most eastern site (with an average infection rate of 22.8% across all species), whereas no rodent was found to be PCR positive in the south-west site and only 6.6% were positive in the north-west of France. Finally, a significant increase in prevalence was observed in autumn samples compared to spring samples in the north-west, but no change was found in the other two sites.

Anaplasma phagocytophilum prevalence in ticks and rodents in an urban and natural habitat in South-Western Slovakia.

BackgroundIxodes ricinus is the principal vector of Anaplasma phagocytophilum, the ethiological agent of granulocytic anaplasmosis in Europe. Anaplasmosis is an emerging zoonotic disease with a natural enzootic cycle. The reservoir competence of rodents is unclear. Monitoring of A. phagocytophilum prevalence in I. ricinus and rodents in various habitat types of Slovakia may contribute to the knowledge about the epidemiology of anaplasmosis in Central Europe.MethodsOver 4400 questing ixodid ticks, 1000 rodent-attached ticks and tissue samples of 606 rodents were screened for A. phagocytophilum DNA by real-time PCR targeting the msp2 gene. Ticks and rodents were captured along six transects in an urban/suburban and natural habitat in south-western Slovakia during 2011–2014. Estimates of wildlife (roe deer, red deer, fallow deer, mouflon, wild boar) densities in the study area were taken from hunter’s yearly reports. Spatial and temporal differences in A. phagocytophilum prevalence in questing I. ricinus and relationships with relative abundance of ticks and wildlife were analysed.ResultsOverall prevalence of A. phagocytophilum in questing I. ricinus was significantly higher in the urban/suburban habitat (7.2 %; 95 % CI: 6.1–8.3 %) compared to the natural habitat (3.1 %; 95 % CI: 2.5–3.9 %) (χ2 = 37.451; P < 0.001). Significant local differences in prevalence of infected questing ticks were found among transects within each habitat as well as among years and between seasons. The trapped rodents belonged to six species. Apodemus flavicollis and Myodes glareolus prevailed in both habitats, Microtus arvalis was present only in the natural habitat. I. ricinus comprised 96.3 % of the rodent-attached ticks, the rest were Haemaphysalis concinna, Ixodes trianguliceps and Dermacentor reticulatus. Only 0.5 % of rodent skin and 0.6 % of rodent-attached ticks (only I. ricinus) were infected with A. phagocytophilum. Prevalence of A. phagocytophilum in questing I. ricinus did not correlate significantly with relative abundance of ticks or with abundance of wildlife in the area.ConclusionThe study confirms that urban I. ricinus populations are infected with A. phagocytophilum at a higher rate than in a natural habitat of south-western Slovakia and suggests that rodents are not the main reservoirs of the bacterium in the investigated area.

Detection ofAnaplasma phagocytophilumin Ixodid Ticks from Equine-Inhabited Sites in the Southeastern United States

Anaplasma phagocytophilum is a vector-borne, obligate intracellular bacterium that invades the neutrophils and eosinophils of infected individuals, causing granulocytic anaplasmosis. Equine cases have previously been reported in the United States from California, Florida, and Connecticut, but limited surveillance studies in the Southeast have been conducted. The objective of this study was to determine A. phagocytophilum prevalence in Ixodes scapularis ticks at southeastern U.S. horse-inhabited sites to evaluate the potential risk for equine exposure to A. phagocytophilum-infected ticks in these areas. Samples of I. scapularis were collected from selected barrier islands and Georgia mainland sites where feral and domestic equine populations are present, respectively. Ticks were individually tested for infection by amplification of the A. phagocytophilum ankA gene. The collective prevalence of A. phagocytophilum in I. scapularis ticks was 20% (n=808).

Detection and identification of Anaplasma phagocytophilum, Borrelia burgdorferi, and Rickettsia helvetica in Danish Ixodes ricinus ticks

Borreliosis is an endemic infection in Denmark. Recent serosurveys have indicated that human anaplasmosis may be equally common. The aim of this study was to look for Anaplasma phagocytophilum and related pathogens in Ixodes ricinus ticks and estimate their prevalence, compared to Borrelia, using PCR. Ticks were collected from three locations in Denmark: Jutland, Funen, and Bornholm. Ticks from Jutland and Funen were analysed individually, ticks from Bornholm were analysed in pools of 20. A. phagocytophilum was found in ticks from all areas. A. phagocytophilum was found in 23.6% of ticks from Jutland and Funen, while 11% were positive for Borrelia burgdorferi. The Borrelia genotype B. afzelii was most prevalent, followed by B. valaisiana, B. burgdorferi s.s. and B. garinii.A. phagocytophilum was found in 14.5% of nymphs and 40.5% of adult ticks, while Borrelia was found in 13% of nymphs and 8% of adult ticks. The difference in prevalence between Anaplasma and Borrelia in adult ticks supports the idea that their maintenance cycles in nature may be different. Ticks were also infected with Rickettsia helvetica. Our study indicates that A. phagocytophilum prevalence in ticks in Denmark is as high as Borrelia prevalence and that human anaplasmosis may be unrecognized.

Ticks and tick-borne disease in Guatemalan cattle and horses

Blood samples and ticks were collected from 48 cattle and 74 horses from seven sites in the Peten region of Guatemala. Data on body condition, mucous membrane capillary refill time and tick infestation levels were recorded for each animal in the study. Horses had significantly higher levels of tick infestation than cattle, as well as poorer body condition scores. Seroprevalence of Babesia spp. was 95.8% for B. bovis in cattle, 89.6% for B. bigemina in cattle, and 92.7% for B. equi in horses. Seroprevalence of Anaplasma marginale in cattle was 87.5%, similar to reports in animals from other regions of Central America. This is the first time that A. phagocytophilum has been reported in animals from this region, with overall PCR-prevalence of 27.6% in cattle and horses, and seroprevalence of 28.4% (52% in cattle and 13% in horses). An agent was identified with serological cross-reactivity and close genetic relatedness to Ehrlichia ruminantium, but further testing confirmed that the agent in Guatemalan cows was not the agent of heartwater. Ticks were identified to species with the predominant species identified on cattle as Boophilus microplus and Amblyomma cajennense, while Anocentor nitens and A. cajennense were most commonly found on horses. Prevalence of infection, tick infestation levels, host factors and environmental data were analyzed for association; A. nitens was significantly associated with A. phagocytophilum prevalence by village.