Italian peninsula as a hybridization zone of Ixodes inopinatus and I. ricinus and the prevalence of tick-borne pathogens in I. inopinatus, I. ricinus, and their hybrids.

BACKGROUND: Ixodes inopinatus was described from Spain on the basis of morphology and partial sequencing of 16S ribosomal DNA. However, several studies suggested that morphological differences between I. inopinatus and Ixodes ricinus are minimal and that 16S rDNA lacks the power to distinguish the two species. Furthermore, nuclear and mitochondrial markers indicated evidence of hybridization between I. inopinatus and I. ricinus. In this study, we tested our hypothesis on tick dispersal from North Africa to Southern Europe and determined the prevalence of selected tick-borne pathogens (TBPs) in I. inopinatus, I. ricinus, and their hybrids. METHODS: Ticks were collected in Italy and Algeria by flagging, identified by sequencing of partial TROSPA and COI genes, and screened for Borrelia burgdorferi s.l., B. miyamotoi, Rickettsia spp., and Anaplasma phagocytophilum by polymerase chain reaction and sequencing of specific markers. RESULTS: Out of the 380 ticks, in Italy, 92 were I. ricinus, 3 were I. inopinatus, and 136 were hybrids of the two species. All 149 ticks from Algeria were I. inopinatus. Overall, 60% of ticks were positive for at least one TBP. Borrelia burgdorferi s.l. was detected in 19.5% of ticks, and it was significantly more prevalent in Ixodes ticks from Algeria than in ticks from Italy. Prevalence of Rickettsia spotted fever group (SFG) was 51.1%, with significantly greater prevalence in ticks from Algeria than in ticks from Italy. Borrelia miyamotoi and A. phagocytophilum were detected in low prevalence (0.9% and 5.2%, respectively) and only in ticks from Italy. CONCLUSIONS: This study indicates that I. inopinatus is a dominant species in Algeria, while I. ricinus and hybrids were common in Italy. The higher prevalence of B. burgdorferi s.l. and Rickettsia SFG in I. inopinatus compared with that in I. ricinus might be due to geographical and ecological differences between these two tick species. The role of I. inopinatus in the epidemiology of TBPs needs further investigation in the Mediterranean Basin.

Detection of Anaplasma phagocytophilum in horses from Germany by molecular and serological testing (2008-2021).

BACKGROUND: Equine granulocytic anaplasmosis (EGA) is a tick-borne disease caused by Anaplasma (A.) phagocytophilum. In Germany, this pathogen is transmitted primarily by Ixodes ricinus. There is limited knowledge about its prevalence in horses in Germany. The aim of this retrospective study was to analyze the results of serological and molecular testing for A. phagocytophilum in horses which were done in a commercial laboratory in Germany over fourteen years. Additionally, risk factors were evaluated, and hematological abnormalities were addressed in horses with positive PCR results. METHODS: This retrospective study examined results of direct (Polymerase chain reaction [PCR]) and indirect (immunofluorescence antibody test [IFAT]) detection methods for A. phagocytophilum in horses on samples provided by German veterinarians and processed by the commercial laboratory LABOKLIN from 2008 to 2021. In horses with positive test results, a Complete Blood Count (CBC) and Serum Amyloid A (SAA) were also analyzed where possible. RESULTS: In total, 1217/4834 horses tested positive (PCR: 190/1246 horses, 15.2%; IFAT: 1036/3849 horses, 26.9%). Seasonality and location, as classified by federal state, had a statistically significant impact on PCR results (P < 0.001 for both). In horses with positive PCR results, hematological abnormalities were detected in 112/118 horses (95%), with thrombocytopenia (86%) and anemia (52%) representing the most common findings. The remaining 6/118 horses (5%) showed no hematological abnormalities on CBC. SAA was measured in 35 horses with positive PCR results, which exclusively showed marked elevation. CONCLUSIONS: The seasonality of A. phagocytophilum infections confirmed by PCR testing was consistent with known peaks in vector activity in Germany. The high rate of horses with positive PCR results when compared to dogs and cats may be due to a lack of ectoparasite prophylaxis. Infections with A. phagocytophilum should be considered as a differential diagnosis in horses with cytopenia on CBC and SAA elevation, especially in the summer and after any possible tick exposure.

Range Expansion of Ixodes scapularis and Borrelia burgdorferi in Ontario, Canada, from 2017 to 2019.

Range expansion of the vector tick species, Ixodes scapularis, has been detected in Ontario over the last two decades. This has led to elevated risk of exposure to Borrelia burgdorferi, the bacterium that causes Lyme disease. Previous research using passive surveillance data suggests that I. scapularis populations establish before the establishment of B. burgdorferi transmission cycles, with a delay of ∼5 years. The objectives of this research were to examine spatial and temporal patterns of I. scapularis and its pathogens from 2017 to 2019 in southwestern, eastern, and central Ontario, and to explore patterns of B. burgdorferi invasion. Over the 3-year study period, drag sampling was conducted at 48 sites across Ontario. I. scapularis ticks were tested for B. burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia species, including Babesia microti and Babesia odocoilei, and Powassan virus. I. scapularis was detected at 30 sites overall, 22 of which had no history of previous tick detection. B. burgdorferi was detected at nine sites, eight of which tested positive for the first time during this study and five of which had B. burgdorferi detected concurrently with initial tick detection. Tick and pathogen hotspots were identified in eastern Ontario in 2017 and 2018, respectively. These findings provide additional evidence on the range expansion and population establishment of I. scapularis in Ontario and help generate hypotheses on the invasion of B. burgdorferi in Ontario. Ongoing public health surveillance is critical to monitor changes in I. scapularis and its pathogens in Ontario.

Seroprevalence of Anaplasma spp. and Ehrlichia spp. and molecular detection of Anaplasma phagocytophilum and Anaplasma platys in stray dogs in Bosnia and Herzegovina.

Stray dogs may be highly exposed to vector-borne pathogens (VBPs), including zoonotic agents, and therefore may pose a high risk of spreading infections to other animals and humans. Among the Anaplasmataceae, Anaplasma phagocytophilum, A. platys and Ehrlichia canis are commonly identified species in dogs in Europe; however, information on the occurrence of these pathogens in canine populations from Bosnia and Herzegovina (B&H) is still lacking. Thus, the aim of this study was to determine the seroprevalence of Anaplasma spp. and Ehrlichia spp. in stray dogs in the Sarajevo region of B&H and to identify A. phagocytophilum, A. platys, E. canis and E. ewingii by molecular techniques. A total of 903 blood samples of stray dogs were screened by SNAP 4Dx Plus Test for the presence of antibodies against A. phagocytophilum/A. platys and E. canis/E. ewingii. Real-time PCR assays were performed for the detection of Anaplasmataceae, A. phagocytophilum, A. platys, E. canis and E. ewingii in seropositive dogs. Antibodies to A. phagocytophilum/A. platys and/or E. canis/E. ewingii were detected in 187 (20.7%) samples. Seroprevalence was highest for A. phagocytophilum/A. platys (184/903, 20.4%). Two dogs had antibodies to E. canis/E. ewingii, while one dog was found to have antibodies to A. phagocytophilum/A. platys and to E. canis/E. ewingii. Forty-eight (25.7%) of the 187 seropositive dogs examined by Real-time PCR were positive for Anaplasmataceae. A. phagocytophilum was detected in 45 (24%) samples, while one sample was positive for A. phagocytophilum and A. platys. Two samples positive for Anaplasmataceae tested negative in the species-specific PCRs. E. canis or E. ewingii could not be detected in any of the Ehrlichia-seropositive dogs. These findings highlight the need for dog health monitoring, improving the health and welfare of stray dog population, and establishment of effective surveillance systems to combat VBDs.

Impact of tick-borne Anaplasma phagocytophilum infections in calves of moose (Alces alces) in southern Norway.

The Gram-negative, obligate intracellular tick-transmitted pathogen Anaplasma phagocytophilum can cause acute febrile diseases in humans and domestic animals. The expansion of the tick Ixodes ricinus (Linnaeus, 1758) in northern Europe due to climate change is of serious concern for animal and human health. The aim of the present study was to investigate the impact of A. phagocytophilum infection in moose Alces alces (Linnaeus) calves by evaluating the carcass weights of infected and non-infected animals and examining animal tissues samples for co-infections with either species of Babesia Starcovici, 1893 or bacteria of the genus Bartonella. The carcasses of 68 free-ranging moose calves were weighed by hunters during the hunting seasons from 2014 to 2017 in two regions in southern Norway and spleen samples were collected. Anaplasma phagocytophilum was detected in moose sampled from locations infected with ticks with a prevalence of 82% (n = 46). The carcass weights of A. phagocytophilum-infected calves (n = 46) and non-infected (n = 22) calves were compared. Although the average weight of infected calves (45.6 kg) was lower than that of non-infected calves (46.5 kg), the difference was not statistically significant. Three different variants of the bacterium 16S rRNA gene were identified. The average weight of animals infected with variant I was 49.9 kg, whereas that of animals infected with variant III was 42.0 kg, but the difference was not statistically significant (p = 0.077). Co-infections of A. phagocytophilum with Bartonella spp. or with Babesia spp. were found in 20 and two calves, respectively. A triple infection was found in two calves. Sequence analysis of the 18S rRNA gene of Babesia-positive samples revealed the presence of Babesia cf. odocoilei (Emerson et Wright, 1970). Strains of Bartonella closely related to Bartonella bovis (Bermond, Boulouis, Heller, Laere, Monteil, Chomel, Sander, Dehio et Piemont, 2002) were identified based on phylogenetic analysis of the gltA and rpoB genes. The loss of body mass in moose calves in the tick-infected site was probably influenced by multiple factors.

Citizen Science Provides an Efficient Method for Broad-Scale Tick-Borne Pathogen Surveillance of Ixodes pacificus and Ixodes scapularis across the United States.

Tick-borne diseases have expanded over the last 2 decades as a result of shifts in tick and pathogen distributions. These shifts have significantly increased the need for accurate portrayal of real-time pathogen distributions and prevalence in hopes of stemming increases in human morbidity. Traditionally, pathogen distribution and prevalence have been monitored through case reports or scientific collections of ticks or reservoir hosts, both of which have challenges that impact the extent, availability, and accuracy of these data. Citizen science tick collections and testing campaigns supplement these data and provide timely estimates of pathogen prevalence and distributions to help characterize and understand tick-borne disease threats to communities. We utilized our national citizen science tick collection and testing program to describe the distribution and prevalence of four Ixodes-borne pathogens, Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti, across the continental United States. IMPORTANCE In the 21st century, zoonotic pathogens continue to emerge, while previously discovered pathogens continue to have changes within their distribution and prevalence. Monitoring these pathogens is resource intensive, requiring both field and laboratory support; thus, data sets are often limited within their spatial and temporal extents. Citizen science collections provide a method to harness the general public to collect samples, enabling real-time monitoring of pathogen distribution and prevalence.

A survey on equine tick-borne diseases: The molecular detection of Babesia ovis DNA in Turkish racehorses.

Common vector-borne diseases of horses include equine piroplasmosis (EP) caused by Babesia caballi and Theileria equi, and equine granulocytic anaplasmosis (EGA) caused by Anaplasma phagocytophilum. Equine piroplasmosis leads to severe health issues in horses and restrictions on the movement of horses internationally. Anaplasma phagocytophilum causes an acute febrile illness in horses and is also of zoonotic importance. In the present study, blood samples were collected from 152 Turkish racehorses from three different provinces (Izmir, Gaziantep, and Konya) of Turkey to investigate the prevalence of EP and EGA. Standard and nested polymerase chain reactions were performed to identify equine piroplasms and A. phagocytophilum, respectively. PCR primers targeting Babesia spp. 18S rRNA, B. caballi BC48, T. equi EMA-1, and A. phagocytophilum 16S rRNA genes were used for molecular diagnosis. Following the cloning and subsequent sequencing of PCR-positive samples, a total of 15 (9.9%) horses were found to be infected with at least one pathogen. Theileria equi and A. phagocytophilum were found in 3.3% (5/152) and 6.6% (10/152) of the samples, respectively. Although B. caballi specimens were not detected in any of the samples, a positive signal was detected for the Babesia genus-specific 18S rRNA PCR. Subsequent sequencing of this signal revealed 100% identity to Babesia ovis. This is the first detection of B. ovis DNA in racehorses in Turkey to the best of our knowledge. Additionally, this study also reports the first molecular identification of A. phagocytophilum in Turkish racehorses. Based on this report, it is recommended that future epidemiological studies on horses also take B. ovis, a parasite usually found in sheep, into consideration and that further detailed studies be conducted to unravel the transmission pathways and potential clinical effects of B. ovis in horses.

Molecular investigation of tick-borne pathogens in ticks removed from tick-bitten humans in the southwestern region of the Republic of Korea.

In this study, we investigated the presence of tick-borne pathogens in ticks removed from tick-bitten humans in the southwestern provinces of the Republic of Korea (ROK). We identified 33 ticks from three tick species, namely Amblyomma testudinarium (60.6%), Haemaphysalis longicornis (27.3%), and Ixodes nipponensis (12.1%) in order of occurrence via morphology and 16S rDNA-targeting polymerase chain reaction (PCR). Tick-borne pathogens were detected in 16 ticks using pathogen-specific PCR. From the results, 12 ticks (36.4%) tested positive for spotted fever group (SFG) Rickettsia: Rickettsia monacensis (1/12), R. tamurae (8/12), and Candidatus Rickettsia jingxinensis (3/12). Three ticks (9.1%) were positive for Anaplasma phagocytophilum. In addition, three ticks (9.1%) tested positive for Babesia gibsoni (1/3) and B. microti (2/3). In conclusion, we identified three tick species; the most common species was A. testudinarium, followed by H. longicornis and I. nipponensis. SFG Rickettsia, A. phagocytophilum, and Babesia spp. were the most frequently detected pathogens in ticks removed from tick-bitten humans. To our knowledge, this is the first report of R. tamurae and Ca. R. jingxinensis detection in Korea. The present results will contribute to the understanding of tick-borne infections in animals and humans in the ROK.

Are other tick-borne infections overlooked in patients investigated for Lyme neuroborreliosis? A large retrospective study from South-eastern Sweden.

In Europe, the hard tick Ixodes ricinus is considered the most important vector of human zoonotic diseases. Human pathogenic agents spread by I. ricinus in Sweden include Borrelia burgdorferi sensu lato (s.l.), Anaplasma phagocytophilum, Rickettsia helvetica, the recently described Neoehrlichia mikurensis, Borrelia miyamotoi, tick-borne encephalitis virus (TBEV), and Babesia spp. (Babesia microti, Babesia venatorum and Babesia divergens). Since these pathogens share the same vector, co-infections with more than one tick-borne pathogen may occur and thus complicate the diagnosis and clinical management of the patient due to possibly altered symptomatology. Borrelia burgdorferi s.l., TBEV and B. miyamotoi are well-known to cause infections of the central nervous system (CNS), whereas the abilities of other tick-borne pathogens to invade the CNS are largely unknown. The aim of this study was to investigate the presence and clinical impact of tick-borne pathogens other than B. burgdorferi s.l. in the cerebrospinal fluid (CSF) and serum samples of patients who were under investigation for Lyme neuroborreliosis (LNB) in a tick-endemic region of South-eastern Sweden. CSF and serum samples from 600 patients, recruited from the Regions of Östergötland County, Jönköping County and Kalmar County in South-eastern Sweden and investigated for LNB during the period of 2009-2013, were retrospectively collected for analysis. The samples were analysed by real-time PCR for the presence of nucleic acid from B. burgdorferi s.l., B. miyamotoi, A. phagocytophilum, Rickettsia spp., N. mikurensis, TBEV and Babesia spp. Serological analyses were conducted in CSF and serum samples for all patients regarding B. burgdorferi s.l., and for the patients with CSF mononuclear pleocytosis, analyses of antibodies to B. miyamotoi, A. phagocytophilum, spotted fever group (SFG) rickettsiae, TBEV and B. microti in serum were performed. The medical charts of all the patients with CSF mononuclear pleocytosis and patients with positive PCR findings were reviewed. Of the 600 patients, 55 (9%) presented with CSF mononuclear pleocytosis, 13 (2%) of whom had Borrelia-specific antibodies in the CSF. One patient was PCR-positive for N. mikurensis, and another one was PCR-positive for Borrelia spp. in serum. No pathogens were detected by PCR in the CSF samples. Four patients had serum antibodies to B. miyamotoi, four patients to A. phagocytophilum, five patients to SFG rickettsiae, and six patients to TBEV. One patient, with antibodies to SFG rickettsiae, had both clinical and laboratory signs suggestive of a current infection. Nine patients had serum antibodies to more than one pathogen, although none of these was assessed as a current co-infection. We can conclude from this study that tick-borne co-infections are uncommon in patients who are being investigated for suspected LNB in South-eastern Sweden, an area endemic for borreliosis and TBE.

Molecular detection of Anaplasma phagocytophilum in hair and spleen of cats revealed a possible underestimation of feline vector-borne pathogens.

Feline Vector-Borne Diseases show increased global prevalence and some Anaplasma and Ehrlichia species may pose a risk to human health. The diagnosis of Anaplasma and Ehrlichia species infection in cats is achieved by the combined use of different methods as cytologic examination evidencing intracytoplasmic morulae, serologic tests and molecular assays. The peripheral whole blood is considered the sample of choice for Anaplasma and Ehrlichia species DNA detection in cats, but false negative results are reported leading to underestimation of infection prevalence. In order to have a more accurate assessment of the spread of feline vector-borne pathogens, the presence of Anaplasma spp. and Ehrlichia spp. DNA in 37 owner and shelter cats subjected to necropsy were prospectively investigated by testing in end-point PCR spleen, bone marrow, blood clot and hair samples. The bacteria identified were genetically characterised. Three shelter cats tested positive for A. phagocytophilum DNA in spleen (one cat) or in hair samples (two cats). None of the cats tested positive in bone marrow and blood samples. From the results obtained, it can be assumed that the use of spleen or hair samples could allow a more reliable detection of A. phagocytophilum DNA in cats with blood tested negative. In the phylogeny constructed with a fragment of the heat shock (groEL) gene nucleotide sequences, all the identified A. phagocytophilum clustered with bacteria infecting a wide range of hosts, including humans, showing a potential zoonotic role.

Monitoring the patterns of submission and presence of tick-borne pathogens in Ixodes scapularis collected from humans and companion animals in Ontario, Canada (2011-2017).

BACKGROUND: The universal nature of the human-companion animal relationship and their shared ticks and tick-borne pathogens offers an opportunity for improving public and veterinary health surveillance. With this in mind, we describe the spatiotemporal trends for blacklegged tick (Ixodes scapularis) submissions from humans and companion animals in Ontario, along with pathogen prevalence. METHODS: We tested tick samples submitted through passive surveillance (2011-2017) from humans and companion animals for Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum and Babesia microti. We describe pathogen prevalence in ticks from humans and from companion animals and constructed univariable Poisson and negative binomial regression models to explore the spatiotemporal relationship between the rates of tick submissions by host type. RESULTS: During the study, there were 17,230 blacklegged tick samples submitted from humans and 4375 from companion animals. Tick submission rates from companion animals were higher than expected in several public health units (PHUs) lacking established tick populations, potentially indicating newly emerging populations. Pathogen prevalence in ticks was higher in PHUs where established blacklegged tick populations exist. Borrelia burgdorferi prevalence was higher in ticks collected from humans (maximum likelihood estimate, MLE = 17.5%; 95% confidence interval, CI 16.97-18.09%) than from companion animals (9.9%, 95% CI 9.15-10.78%). There was no difference in pathogen prevalence in ticks by host type for the remaining pathogens, which were found in less than 1% of tested ticks. The most common co-infection B. burgdorferi + B. miyamotoi occurred in 0.11% of blacklegged ticks from humans and animals combined. Borrelia burgdorferi prevalence was higher in unengorged (21.9%, 95% CI 21.12-22.65%) than engorged ticks (10.0%, 95% CI 9.45-10.56%). There were no consistent and significant spatiotemporal relationships detected via regression models between the annual rates of submission of each host type. CONCLUSIONS: While B. burgdorferi has been present in blacklegged ticks in Ontario for several decades, other tick-borne pathogens are also present at low prevalence. Blacklegged tick and pathogen surveillance data can be used to monitor risk in human and companion animal populations, and efforts are under consideration to unite surveillance efforts for the different target populations.

Development of a real-time PCR method for rapid diagnosis of canine babesiosis and anaplasmosis.

BACKGROUND: Canine babesiosis and anaplasmosis, caused by Babesia canis and Anaplasma phagocytophilum, respectively, are significant tick-borne diseases in Baltic countries. Both diseases can be diagnosed on the basis of clinicopathological findings, by direct pathogen detection in blood smears or by indirect pathogen detection; however, because of high selectivity and specificity, molecular methods may be advantageous. The goal of this study was to develop a duplex real-time polymerase chain reaction (RT-PCR) method for the detection of B. canis and A. phagocytophilum in canine clinical samples. METHODS: Sequence-based polymorphism analysis of genes encoding B. canis-specific merozoite surface protein Bc28.1 (Bc28.1) and A. phagocytophilum malate dehydrogenase (mdh) was performed on pathogen isolates present in Latvian domestic dogs. The obtained results were used to design a species-specific duplex RT-PCR assay. RESULTS: The presence of three B. canis Bc28.1 gene sequence types was revealed in canine samples with a nonuniform geographical distribution, and two types of A. phagocytophilum mdh genes were detected. The novel duplex RT-PCR assay provided correct classification of samples positive and negative for B. canis and A. phagocytophilum. The analytical sensitivity of this assay was ten gene copies/ reaction for both pathogens. CONCLUSIONS: A novel duplex RT-PCR molecular method was developed for the detection of B. canis and A. phagocytophilum in canine clinical samples. Sequence variability of Bc28.1 and mdh genes indicated the genetic variability of B. canis and A. phagocytophilum isolates occurring in Latvian domestic dogs.

Examining Prevalence and Diversity of Tick-Borne Pathogens in Questing Ixodes pacificus Ticks in California.

Tick-borne diseases in California include Lyme disease (caused by Borrelia burgdorferi), infections with Borrelia miyamotoi, and human granulocytic anaplasmosis (caused by Anaplasma phagocytophilum). We surveyed multiple sites and habitats (woodland, grassland, and coastal chaparral) in California to describe spatial patterns of tick-borne pathogen prevalence in western black-legged ticks (Ixodes pacificus). We found that several species of Borrelia-B. burgdorferi, Borrelia americana, and Borrelia bissettiae-were observed in habitats, such as coastal chaparral, that do not harbor obvious reservoir host candidates. Describing tick-borne pathogen prevalence is strongly influenced by the scale of surveillance: aggregating data from individual sites to match jurisdictional boundaries (e.g., county or state) can lower the reported infection prevalence. Considering multiple pathogen species in the same habitat allows a more cohesive interpretation of local pathogen occurrence. IMPORTANCE Understanding the local host ecology and prevalence of zoonotic diseases is vital for public health. Using tick-borne diseases in California, we show that there is often a bias to our understanding and that studies tend to focus on particular habitats, e.g., Lyme disease in oak woodlands. Other habitats may harbor a surprising diversity of tick-borne pathogens but have been neglected, e.g., coastal chaparral. Explaining pathogen prevalence requires descriptions of data on a local scale; otherwise, aggregating the data can misrepresent the local dynamics of tick-borne diseases.

One man, three tick-borne illnesses.

A 70-year-old man presented to the emergency department with fevers, ankle edema and nausea following a presumed insect bite on his ankle 1 month prior. On examination, he was febrile and had left leg pain with passive range of motion. Laboratory studies revealed anemia, thrombocytopenia, acute kidney injury and elevated aminotransaminases. Due to his recent travel to the Northeastern United States, he was suspected of having a possible tick-borne illness. Serologies were positive for Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti, and the patient was diagnosed with Lyme disease, babesiosis and anaplasmosis. He was treated with doxycyline, atovaquone and azithromycin, leading to resolution of symptoms. While co-infection with Lyme disease is common, infection with three tickborne illnesses at one time is relatively rare.

Prevalence and distribution of Anaplasma phagocytophilum in ticks collected from dogs in the United Kingdom.

BACKGROUND: Anaplasma phagocytophilum is the etiological agent of canine granulocytic anaplasmosis in dogs and causes human granulocytic anaplasmosis (HGA). Tick-borne anaplasmosis has been recognised as an emerging zoonotic health concern worldwide. The aim of the present study was to determine the prevalence of A. phagocytophilum in ticks collected from dogs in the UK and map its distribution. Routine surveillance of tick-borne disease is essential as part of a "One Health" approach to infectious disease management. METHODS: Tick DNA samples collected in 2015 as part of a large-scale tick surveillance programme were analysed using a previously validated diagnostic quantitative PCR for A. phagocytophilum. RESULTS: PCR analysis indicated that 138 out of 2994 tick DNA samples analysed were positive for A. phagocytophilum, a prevalence of 4.6% (95% CI: 3.89-5.42). Among these 138 tick DNA samples, 131 were from Ixodes ricinus, six were from Ixodes hexagonus and one was from Ixodes canisuga. Three of the I. ricinus tick DNA samples positive for A. phagocytophilum DNA were also positive for Borrelia spp. DNA and one was positive for Babesia spp. DNA, indicating co-infection. The ticks positive for the pathogen DNA were found widely distributed throughout the UK. CONCLUSIONS: These data provide important information on the prevalence and wide distribution of A. phagocytophilum in ticks infesting dogs within the UK.

Granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland.

BACKGROUND: Anaplasma are obligate intracellular bacteria and aetiological agents of tick-borne diseases of both veterinary and medical interest. The genus Anaplasma comprises six species: Anaplasma marginale, Anaplasma centrale, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma bovis and Anaplasma platys. They can infect humans, carnivores, ruminants, rodents, insectivores, birds and reptiles. The aim of this study was to present the first clinical case of granulocytic anaplasmosis in a captive ring-tailed lemur in Poland. CASE PRESENTATION: A 4-year-old female lemur presented anorexia, epistaxis and tick infestation. The microscopic examination of a blood smear revealed morulae in neutrophils. Polymerase chain reaction test and sequencing of obtained PCR product confirmed infection by the GU183908 Anaplasma phagocytophilum strain. Therapeutic protocol included doxycycline (2.5 mg/kg p.o., b.i.d.) for 3 weeks and the lemur recovered within 24 h. CONCLUSIONS: This is the first report on granulocytic anaplasmosis in a ring-tailed lemur in Europe, indicating that A. phagocytophilum infection must also be considered in differential diagnosis in this animal species, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.

The potential risk of exposure to Borrelia garinii, Anaplasma phagocytophilum and Babesia microti in the Wolinski National Park (north-western Poland).

Ixodes ricinus (Acari: Ixodida) is the main vector in Europe of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. Wolinski National Park (WNP) is situated by the Baltic Sea and is frequently visited by tourists. The aim of the study was to determine the potential risk of exposure to tick borne infection with B. burgdorferi s.l., A. phagocytophilum and B. microti on the areas of WNP. In total, 394 I. ricinus were tested. The pathogens in ticks were detected by PCR, nested PCR, RFLP and sequencing. Altogether, pathogens were detected in 12.69% of the studied ticks. B. burgdorferi s.l., was shown in 0.25% of the studied I. ricinus, while A. phagocytophilum and B. microti were detected in 1.01% and 10.65% of studied ticks, respectively. Co-infection by A. phagocytophilum and B. microti was shown in only one I. ricinus nymph. Analysis of B. burgdorferi s.l., genospecies showed that 0.25% of the studied ticks were infected with Borrelia garinii. The obtained results show the potentially high human risk of exposure to tick-borne infection with B. microti, and the low potential risk of infection with B. garinii and A. phagocytophilum on the studied areas of WNP.

Babesia spp. and Anaplasma phagocytophilum in free-ranging wild ungulates in central Austria.

Free-ranging wild ungulates are widespread in Austria, and act as hosts (i.e. feeding hosts) for ticks, including Ixodes ricinus, and as reservoir hosts for pathogens transmitted by I. ricinus. Due to climate change, the abundance of I. ricinus might be increasing, which could potentially lead to higher prevalences of tick-borne pathogens, such as Babesia spp. and Anaplasma phagocytophilum, some known for their zoonotic potential. Human babesiosis is classified as an emerging zoonosis, but sufficient data of these parasites in central Austria is lacking. In order to assess the abundance of vector-borne pathogens, blood of roe deer (Capreolus capreolus; n = 137), red deer (Cervus elaphus; n = 37), mouflons (Ovis gmelini; n = 2) and chamois (Rupicapra rupicapra; n = 1), was collected and tested for pathogen DNA in two different sampling sites in central Austria. DNA of tick-borne pathogens was detected in 15.5 % (n = 27) of these animals. Babesia capreoli (n = 22 in roe deer; n = 1 in mouflon), Babesia divergens (n = 1, in red deer), and Anaplasma phagocytophilum (n = 4, in roe deer) were detected. DNA sequencing of the 18S rRNA gene of two C. capreolus samples from Upper Austria featured another new genotype of Babesia, which differs in one nucleotide position to B. divergens and B. capreoli, and is intermediate between the main genotypes of B. capreoli and B. divergens within the partial gene sequence analyzed. This study thus confirms that B. capreoli, B. divergens, and A. phagocytophilum are present in free-ranging ungulates in central Austria. Further testing over a longer period is recommended in order to assess the impact of climate change on the prevalence of blood parasites in central Austria.

Molecular surveillance of tick-borne diseases affecting horses in Poland-Own observations.

The purpose of the study was to carry out the molecular surveillance of piroplasmosis, granulocytic anaplasmosis and lyme borreliosis in horses which originated from Poland and exhibited symptoms raising the suspicion of the aforementioned disease units. The presence of Theileria equi genetic material was detected in 37 out of 512 examined horses (7.2%), and Anaplasma phagocytophilum in 9 (1.8%). The DNA of Borrelia burgdorferi was found in 11 out of 204 examined horses (5.4%). The above-cited results indicate that the problem of tick-borne diseases affecting horses in Poland is not as significant as in other parts of Europe, however they have to be considered in differential diagnosis of the diseases with lethargy, fever, anaemia and thrombocytopenia.