Spotted fever group rickettsiae identified in Dermacentor marginatus and Ixodes ricinus ticks in Algeria

Our study was carried out using Ixodes ricinus ticks collected from cattle from Tizi-Ouzou and Dermacentor marginatus ticks collected from the vegetation of the Blida region, a tourist site, both regions situated in northern Algeria. The results of real-time quantitative PCR (qPCR) specific for a partial sequence of the citrate synthase gene (gltA) indicate that Rickettsia spp. were present in 11/23 (48%) and 4/9 (44%) of the examined ticks from Tizi-Ouzou and Blida, respectively. The sequences of Rickettsia helvetica and Ri. monacensis were found in I. ricinus ticks using gltA primers. In addition, Ri. slovaca was detected based on the sequences of the gltA and the outer membrane protein (OmpA) genes in D. marginatus ticks. DNA sequencing to identify the species revealed for the first time the presence of Ri. helvetica in I. ricinus ticks and Ri. slovaca in D. marginatus ticks from Algeria and confirmed the presence of Ri. monacensis.     

Tick-borne zoonotic pathogens in ticks feeding on the common nightingale including a novel strain of Rickettsia sp.

We examined 77 Ixodes ricinus ticks found on 33 out of 120 common nightingales (Luscinia megarhynchos) sampled in the Czech Republic in 2008 for the presence of Borrelia spirochetes, Anaplasma phagocytophilum, Rickettsia spp., and Babesia spp. We detected Borrelia garinii (in 4% of ticks), A. phagocytophilum (1%), Rickettsia helvetica (3%), a novel strain of Rickettsia sp. (sister taxon of R. bellii; 1%), and Babesia sp. EU1 (1%). Thus, we conclude that nightingales are unlikely to be important reservoir hosts for tick-borne pathogens.     

Rickettsia spp. in Ticks, Poland

Ticks are recognized as the main vectors and reservoirs of spotted fever group rickettsiae. We searched for the most prevalent Rickettsia spp. in Poland and found R. slovaca and R. helvetica bacteria in ticks in southern and central Poland; R. raoulti was found in ticks in all parts of Poland.     

Development of three quantitative real-time PCR assays for the detection of Rickettsia raoultii,Rickettsia slovaca,and Rickettsia aeschlimannii and their validation with ticks from the country of Georgia and the Republic of Azerbaijan

A previous surveillance study of human pathogens within ticks collected in the country of Georgia showed a relatively high infection rate for Rickettsia raoultii, R. slovaca, and R. aeschlimannii. These 3 spotted fever group rickettsiae are human pathogens: R. raoultii and R. slovaca cause tick-borne lymphadenopathy (TIBOLA), and R. aeschlimannii causes an infection characterized by fever and maculopapular rash. Three quantitative real-time polymerase chain reaction (qPCR) assays, Rraoul, Rslov, and Raesch were developed and optimized to detect R. raoultii, R. slovaca, and R. aeschlimannii, respectively, by targeting fragments of the outer membrane protein B gene (ompB) using species-specific molecular beacon or TaqMan probes. The 3 qPCR assays showed 100% specificity when tested against a rickettsiae DNA panel (n = 20) and a bacteria DNA panel (n = 12). The limit of detection was found to be at least 3 copies per reaction for all assays. Validation of the assays using previously investigated tick nucleic acid preparations, which included Rickettsia-free tick samples, tick samples that contain R. raoultii, R. slovaca, R. aeschlimannii, and other Rickettsia spp., gave 100% sensitivity for all 3 qPCR assays. In addition, a total of 65 tick nucleic acid preparations (representing 259 individual ticks) collected from the country of Georgia and the Republic of Azerbaijan in 2009 was tested using the 3 qPCR assays. R. raoultii, R. slovaca, and R. aeschlimannii were not detected in any ticks (n = 31) from the Republic of Azerbaijan, but in the ticks from the country of Georgia (n = 228) the minimal infection rate for R. raoultii and R. slovaca in Dermacentor marginatus was 10% and 4%, respectively, and for R. aeschlimannii in Haemaphysalis sulcata and Hyalomma spp. it was 1.9% and 20%, respectively.     

Prevalence of Rickettsia spp., Anaplasma phagocytophilum,and Coxiella burnetii in adult Ixodes ricinus ticks from 29 study areas in central and southern Sweden

A total of 887 adult Ixodes ricinus ticks (469 females and 418 males) from 29 different localities in Sweden were screened for Rickettsia, Anaplasma, and Coxiella DNA using PCR and then subjected to gene sequencing. Rickettsial DNA was detected in 9.5–9.6% of the ticks. Most of the positive ticks were infected with Rickettsia helvetica. One tick harbored another spotted fever rickettsia, closely related to or identical with R. sibirica not previously found in I. ricinus nor in Sweden. Six of the ticks (0.7%) were infected with an Anaplasma sp., presumably A. phagocytophilum. Coxiella burnetii DNA was not detected in any of the ticks. The detection of R. helvetica and A. phagocytophilum in several of the localities sampled suggests that these potentially human-pathogenic agents are common in Sweden.     

Detection of Rickettsia helvetica in Ixodes ricinus ticks collected from Pyrenean chamois in France

Seventy-one Ixodes ricinus ticks collected from Pyrenean chamois (Rupicapra pyrenaica) in the French Pyrenees were tested by real-time polymerase chain reaction to detect the presence of Rickettsia and Bartonella. Four ticks (6%) were positive for R. helvetica. The chamois carries infected ticks, and this enables the dissemination throughout the environment with this bacterium, a potential human pathogen.     

Rickettsia spp. in ticks (Acari: Ixodidae) from wild birds: First detection of Candidatus Rickettsia vini in Hesse,Germany

Tick-borne diseases are recognized as a growing public health concern and cause significant issues in humans and animals by serving as dispersal agents of ticks and their associated pathogens. Birds contribute to the spread of ticks and tick-borne diseases, with migratory birds playing a role in regional dispersal. Only a few studies have been carried out in Germany so far to investigate tick fauna on birds and the pathogens they harbour. In 2017 and 2018, we collected ticks from a total of 358 birds in the northern region of the Federal State of Hesse, Germany. Most ticks were Ixodes ricinus larvae and nymphs (95.4%). However, two ornithophilic species were also collected: Ixodes frontalis (larvae, nymphs and females) and Ixodes arboricola (larvae and nymphs). Ticks were tested individually or in pools, depending on the tick life stage and species for each bird host, in order to detect tick-borne encephalitis (TBE) virus and Rickettsia spp. All samples tested negative for TBE virus. Of 684 investigated tick pools, 162 tested positive for Rickettsia spp. by screening PCR. Of these, 117 carried Rickettsia helvetica detected by specific real-time PCR. The samples which were negative in the R. helvetica PCR were further investigated using five targets (gltA, 16S, ompA IV, ompB, 23S-5S intergenic spacer region) using multi locus sequence typing and phylogenetic analysis. The obtained sequences showed strong phylogenetic relationship to Candidatus Rickettsia vini, which is most closely related to R. japonica and R. heilongjiangensis, and occur in I. arboricola. Three more sequences were determined as R. helvetica in I. ricinus and I. frontalis. This is the first detection of Ca. R. vini in Germany and the first 23S-5S data published for Ca. R. vini, a useful target for Rickettsia species identification.     

Seasonal and habitat variation in the prevalence of Rickettsia helvetica in Ixodes ricinus ticks from Denmark

A total of 704 unfed ticks of the species Ixodes ricinus collected in Denmark were screened for Rickettsia DNA by a genus-specific real-time PCR. Of the nymphs, 4.7% (31/662) were positive for rickettsial DNA. Among the positive ticks, we observed a seasonal and habitat variation. The infection rate was highest in May as compared to July, August, and October. Ecotone (high tick density) showed an elevated prevalence as compared to spruce or beech forests. Sequencing revealed only DNA from R. helvetica.     

The potential role of migratory birds in transmission cycles of Babesia spp., Anaplasma phagocytophilum,and Rickettsia spp.

Babesia spp., Anaplasma phagocytophilum, and Rickettsia spp. are potentially emerging tick-borne pathogens, whereas many issues about their ecology, e.g. reservoir host specificity, are still unclear. In spring 2007, we collected 191 feeding Ixodes ricinus ticks from 99 birds of 11 different species on a German bird conservation island in the Baltic Sea. Babesia spp. were detected in 4.7% (9/191), A. phagocytophilum was present in 2.6% (5/191), and Rickettsia spp. were identified in 7.3% (14/191) of the investigated ticks. Further characterization of Babesia spp. infections resulted in B. divergens and B. microti. Among the Rickettsia spp. infections, we identified at least 2 different species: R. monacensis and R. helvetica. Furthermore, 2 ticks harboured mixed infections. Our study provides first interesting insights into the role of migratory birds in the distribution of several emerging tick-borne pathogens.     

Molecular identification of tick-borne pathogens (Rickettsia spp., Anaplasma phagocytophilum,Borrelia burgdorferi sensu lato,Coxiella burnetii and piroplasms) in questing and feeding hard ticks from North-Western Spain

The occurrence of tick-borne pathogens (TBPs) of human and veterinary interest was studied in questing and feeding ticks collected from wild animals in a region in North-Western Spain. A total of 529 ticks (489 questing, 40 feeding) of seven different species (386 Ixodes ricinus, 53 Haemaphysalis concinna, 27 Haemaphysalis punctata, 25 Dermacentor marginatus, 21 Haemaphysalis inermis, 15 Dermacentor reticulatus, and two Rhipicephalus bursa) were analyzed. Molecular analysis of the 16S rRNA gene in I. ricinus ticks, revealed the presence of two phylogenetic groups in the region. Most of the sequenced ticks (96%) were assigned to I. ricinus haplogroup and 4% of the ticks were phylogenetically related to I. inopinatus haplogroup. Feeding ticks were removed from 17 animals from seven wild species (seven roe deer -Capreolus capreolus-, three wolves -Canis lupus-, two Iberian red deer -Cervus elaphus hispanicus-, two European wild boar -Sus scrofa-, one Cantabrian brown bear -Ursus arctos-, one Eurasian badger -Meles meles-, and one red fox -Vulpes vulpes-). Presence of Rickettsia spp., Anaplasma phagocytophilum, piroplasms, Borrelia burgdorferi sensu lato (s.l.) and Coxiella burnetii were tested in ticks by specific PCR. A total of 92 (17.4%) of the 529 ticks analyzed were positive for at least one of the TBPs tested. Sequencing revealed the presence of the genospecies “Candidatus Rickettsia rioja”, Rickettsia raoultii, and Anaplasma phagocytophilum in both questing and feeding ticks. Rickettsia slovaca, Borrelia lusitaniae, Borrelia afzelii, Borrelia garinii, Borrelia burgdorferi sensu stricto and Babesia bigemina were only detected in questing ticks, while Babesia sp. badger type A, Theileria OT3 and Hepatozoon canis occurred only in engorged ticks. None of the ticks were positive for C. burnetii. The analysis of the 16S rRNA gene sequences of A. phagocytophilum revealed the presence of three variants (I, X and W) circulating in the region. New host-tick-pathogen interactions have been revealed, finding for the first time the human pathogen R. raoultii in D. reticulatus removed from a Cantabrian brown bear. Co-occurrence between different TBPs were detected in 4.3% of the ticks. The association B. burgdorferi s.l./Rickettsia spp. was detected in questing ticks; and Rickettsia spp./piroplasms and A. phagocytophilum/Theileria OT3 in feeding ticks. The presence of pathogenic agents constitutes a threat to human and animal health, and should be considered in the diagnosis and treatment after a tick bite. This study increases the knowledge on TBPs diversity of medical and veterinary interest circulating between ticks and their hosts in North-Western Spain.     

Prevalence of Coxiella burnetii and Rickettsia spp. in ticks and rodents in southern Germany

Coxiella burnetii, the causative agent of Q fever, and Rickettsia spp. are bacterial pathogens that can be transmitted by ticks of the genus Dermacentor (i.e., Dermacentor marginatus and D. reticulatus). In Germany, the occurrence of these ticks is currently limited to few areas. However, due to increasing temperatures, these vectors will likely extend their distribution in the future, and C. burnetii and Rickettsia spp. might spread with them. To assess the prospective risk of human infections by these agents, it is important to know their current distribution. We collected 666 adult Dermacentor spp. and 119 rodents, mainly Microtus arvalis, in 3 Q fever endemic areas in southern Germany. Ticks and rodent organ pools were screened by PCR for C. burnetii and Rickettsia spp. No evidence of C. burnetii infections could be found in ticks or rodents, suggesting that these animals do not play an essential role in the epidemiology of Q fever in Germany. Rickettsia raoultii and R. slovaca could be detected in 30.3% and 0.75% of all examined ticks, respectively. In contrast, no rickettsia infections could be found in any rodent samples. Both rickettsia species can cause tick-borne lymphadenopathy (TIBOLA), a usually mild human disease. Because of the possible transmission of these rickettsiae to humans, TIBOLA should be considered in the differential diagnosis of tick-borne diseases. Our data show that a spread of these rickettsiae is possible in Germany and that more studies on the distribution of these agents are necessary.     

Detection of Leishmania infantum,Babesia canis,and rickettsiae in ticks removed from dogs living in Italy

The aims of this study were to determine natural infections by Anaplasma phagocytophilum/Anaplasma platys, Bartonella henselae, Ehrlichia canis, Leishmania infantum, Rickettsia spp., Babesia spp., and Hepatozoon spp. by molecular methods in ticks (n = 91) removed from dogs with clinical signs and laboratory abnormalities compatible with tick-borne diseases (n = 22) living in Italy and to assess the distribution and species of ticks encountered. Ticks from dogs living in southern Italy were all identified as Rhipicephalus sanguineus (n = 25), ticks from central Italy included Rh. sanguineus (n = 8) and Ixodes ricinus (n = 9), ticks from northern Italy included Rh. sanguineus (n = 45), Dermacentor marginatus (n = 3), and one I. ricinus. Leishmania infantum, Rickettsia spp., and Babesia canis were the only pathogens detected in 7 (8%), 4 (4%), and 2 (2%) out of 91 ticks, respectively. L. infantum was detected in I. ricinus from central Italy and in Rh. sanguineus from northern and central Italy. Rickettsia conorii and Ri. massiliae were detected in Rh. sanguineus ticks from central and southern Italy (Sicily), respectively. Bab. canis was detected in D. marginatus ticks from northern Italy.     

Detection of Rickettsia spp. and Anaplasma phagocytophilum in Ixodes ricinus ticks in a region of Middle Germany (Thuringia)

Rickettsia spp. and Anaplasma spp. are regarded as potentially emerging tick-borne pathogens, but so far data on prevalence rates in questing ticks and reports on human diseases in several parts of Europe are rarely available.In this study, 430 nymphs and 570 adult Ixodes (I.) ricinus ticks were collected from a frequently visited forest region of Thuringia (Zeitzgrund, near Stadtroda) in 2006 (n=506) and 2007 (n=494). Individual ticks were investigated for a part of the citrate synthase gene (gltA) of Rickettsia spp. and the 16S rRNA gene of Anaplasma phagocytophilum. Positive amplicons were identified with restriction fragment length polymorphism (RFLP) and/or sequencing. Overall, 14.7% (147/1000) of investigated ticks were infected with Rickettsia spp. After sequencing of 64/147 positive amplicons R. helvetica (29/64) was detected predominantly. Prevalence varied in different developmental stages between 9.3% (40/430) in nymphs and 18.8% (107/570) in adults. A. phagocytophilum-specific DNA was detected in 5.4% (54/1000) of ticks with an infection rate of 4.7% (20/430) in nymphs and 6.0% (34/570) in adults. In 1% (10/1000) of ticks coinfections with Rickettsia spp. and A. phagocytophilum were found. Our study provides interesting insights into the circulation and cocirculation of different rickettsial species and A. phagocytophilum in the same biotope.     

Distribution and prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks of canton Ticino (Switzerland)

Free-living Ixodes ricinus ticks were collected from 12 different sites of canton Ticino, south of the Alps (Switzerland). Each tick was examined for the presence of Borrelia burgdorferi sensu lato (sl), the etiologic agent of Lyme borreliosis using direct fluorescent antibody assay, and isolation of the bacteria. Borreliae were characterized by PCR followed by RFLP. The abundance and infection rates of I. ricinus ticks varied greatly between the areas. Two localities were found free of Borrelia. The prevalence of infected ticks ranged from 5 to 19%. Most ticks (96%) were found infected by <50 spirochetes. Three B. burgdorferi sl species were successfully isolated: B. garinii dominated, followed by B. lusitaniae and B. valaisiana. Additionally, a mixed infection with B. garinii and B. valaisiana was observed. The distribution of the various Borrelia species in the different areas was heterogeneous. This is the first report of the presence of B. lusitaniae in I. ricinus in Switzerland.     

Ticks and tick-borne rickettsiae from dogs in El Salvador,with report of the human pathogen Rickettsia parkeri

Twelve tick species have been reported in El Salvador; however, information regarding ticks infesting domestic dogs is lacking, and pathogenic tick-borne Rickettsia species have never been reported in El Salvador. This work evaluated ticks infesting 230 dogs from ten municipalities in El Salvador from July 2019 to August 2020. A total of 1,264 ticks were collected and identified into five species: Rhipicephalus sanguineus sensu lato (s.l.), Rhipicephalus microplus, Amblyomma mixtum, Amblyomma ovale, and Amblyoma cf. parvum. The tick R. sanguineus s.l. was the most frequent species in all localities (81.3% of sampled dogs), followed by Amblyomma mixtum (13.0%), Amblyomma ovale (10.9%) and Amblyomma cf. parvum (10.4%). The overall mean intensity of tick infestation was 5.5 ticks/dog. The highest specific mean intensity value was for R. sanguineus s.l. (4.8 ticks/dog), varying from 1.6 to 2.7 ticks/dog for the three Amblyomma species. From a random sample of 288 tick specimens tested molecularly for the presence of rickettsial agents, three spotted fever group Rickettsia were detected: Rickettsia amblyommatis in 90% (36/40) A. mixtum, 46% (11/24) A. cf. parvum, 4% (7/186) R. sanguineus s.l., and 17% Amblyomma spp.; Rickettsia parkeri strain Atlantic rainforest in 4% (1/25) A. ovale; and an unnamed rickettsia agent, designated as ‘Rickettsia sp. ES-A.cf.parvum’, in 4% (1/24) A. cf. parvum. Our finding of R. parkeri strain Atlantic rainforest in A. ovale is highly relevant because this agent has been associated to spotted fever illness in other Latin American countries, where A. ovale is implicated as its main vector. These findings suggest that spotted fever cases caused by R. parkeri strain Atlantic rainforest could be occurring in El Salvador.     

Rickettsia slovaca from Dermacentor marginatus ticks in Sardinia,Italy

Nineteen ticks belonging to the species Dermacentor marginatus, Rhipicephalus bursa, and Haemaphysalis sulcata were collected from wild animals (wild boar, deer, and mouflon) in south-western Sardinia, Italy. Five D. marginatus ticks from wild boar were PCR-positive when analyzed using gltA-specific and ompA-specific primers, leading to the identification and first isolation in cell culture of Rickettsia slovaca, the causative agent of tick-borne lymphadenopathy (TIBOLA), on the island of Sardinia. This study confirms the detection of a new tick-borne rickettsia that can be added to the others already known to be present in Sardinia (Rickettsia aeschlimannii, R. massiliae, and Candidatus Rickettsia barbariae). These data increase our knowledge of tick-borne rickettsioses in Sardinia and, more generally, in the Mediterranean basin.     

Rickettsia slovaca Infection in Humans,Portugal

Fifteen years after the initial detection of Rickettsia slovaca in ticks in Portugal, 3 autochthonous cases of R. slovaca infection were diagnosed in humans. All patients had an eschar on the scalp and lymphadenopathy; 2 patients had facial edema. R. slovaca infection was confirmed by serologic testing, culture, and PCR.     

Molecular detection of Rickettsia amblyommatis and Rickettsia parkeri in ticks collected from wild pigs in Campeche,Mexico

Tick-borne rickettsioses are caused at least by 15 species of Rickettsia of the Spotted fever group, which represent a major emerging and re-emerging public health problem worldwide. Some of these microorganisms have complex cycles involving the interaction of multiple species of ticks and wild and domestic mammals. Rickettsia infection was investigated in ticks collected from wild pigs at six localities in southeastern Mexico. We collected and tested 196 ticks belonging to four species, including Amblyomma maculatum, Amblyomma mixtum, Amblyomma ovale and Riphicephalus microplus, from 13 of 20 (65%) wild pigs sampled. Overall, Rickettsia DNA was detected in 13.8% of ticks tested (10 ♂ and 17 ♀). Of the 27 Rickettsia-positive ticks, six were A. maculatum, and 21 A. mixtum. Sequencing and phylogenetic analysis of the gltA and ompB genes revealed the presence of Rickettsia parkeri sensu stricto in one female A. maculatum and Rickettsia amblyommatis in five A. maculatum (2 ♂, 3 ♀) and 21 A. mixtum ticks (8 ♂, 13 ♀). The finding of two rickettsial agents in ticks collected from a wild pig population that is regularly captured and kept in captivity or hunted as a source of food raises concern about potential disease transmission to humans and domestic animals. However, more investigations are needed to further understand the ecology of Rickettsia species in free-ranging animals and their implications for human health.     

Urban family cluster of spotted fever rickettsiosis linked to Rhipicephalus sanguineus infected with Rickettsia conorii subsp. caspia and Rickettsia massiliae

Here, we report an epidemiological and entomological investigation of a cluster of cases of spotted fever group (SFG) rickettsiosis occurring in southern France. A family of 3 (husband, wife, and their son) presented with symptoms compatible with SFG rickettsiosis. For 2 patients, serum samples presented increased levels of IgM and IgG for SFG Rickettsia. The patients’ home was investigated, and Rhipicephalus sanguineus ticks were collected from the floor from behind the furniture. Of 22 ticks collected, 20 tested positive for Rickettsia. As Rh. sanguineus serves as a vector for both Rickettsia conorii and Ri. massiliae in southern France, all Rh. sanguineus isolates were tested by real-time PCR and conventional PCR to detect the 2 species. Nine ticks tested positive for Ri. conorii subsp. caspia (marking the first documentation of this subspecies in France), 7 tested positive for Ri. massiliae, and 4 tested positive for both rickettsiae. This study is the first report of coinfection of Rh. sanguineus ticks with Ri. conorii and Ri. massiliae in southern France.     

Prevalence of Anaplasma phagocytophilum,Borrelia burgdorferi sensu lato,Rickettsia spp. and Babesia spp. in cattle serum and questing ticks from Belgium

BackgroundAnaplasmosis, borreliosis, rickettsiosis and babesiosis are tick-borne diseases of medical, veterinary and economic importance. In Belgium, little is known on the prevalence of these diseases in animals and previous screenings relate only to targeted geographic regions, clinical cases or a limited number of tested samples. We therefore performed the first nationwide seroprevalence study of Anaplasma spp., A. phagocytophilum, Borrelia spp., Rickettsia spp. and Babesia spp. in Belgian cattle. We also screened questing ticks for the aforementioned pathogens.MethodsELISAs and IFATs were performed on a representative sample set of cattle sera stratified proportionally to the number of cattle herds per province. Questing ticks were collected in areas where the highest prevalence for the forenamed pathogens in cattle serum were observed. Ticks were analyzed by quantitative PCR for A. phagocytophilum (n = 783), B. burgdorferi sensu lato (n = 783) and Rickettsia spp. (n = 715) and by PCR for Babesia spp. (n = 358).ResultsThe ELISA screening for antibodies to Anaplasma spp. and Borrelia spp. in cattle sera showed an overall seroprevalence of 15.6% (53/339) and 12.9% (52/402), respectively. The IFAT screening for antibodies against A. phagocytophilum, Rickettsia spp. and Babesia spp. resulted in an overall seroprevalence of 34.2% (116/339), 31.2% (99/317) and 3.4% (14/412), respectively. At the provincial level, the provinces of Liege and Walloon Brabant harboured the highest seroprevalence of Anaplasma spp. (44.4% and 42.7% respectively) and A. phagocytophilum (55.6% and 71.4%). East Flanders and Luxembourg exhibited the highest seroprevalence of Borrelia spp. (32.4%) and Rickettsia spp. (54.8%) respectively. The province of Antwerp showed the highest seroprevalence of Babesia spp. (11%). The screening of field-collected ticks resulted in a prevalence of 13.8% for B. burgdorferi s.l., with B. afzelii and B. garinii being the most common genospecies (65.7% and 17.1%, respectively). Rickettsia spp. was detected in 7.1% of the tested ticks and the only identified species was R. helvetica. A low prevalence was found for A. phagocytophilum (0.5%) and no Babesia positive tick was detected.ConclusionsThe seroprevalence data in cattle indicate hot spots for tick-borne pathogens in specific provinces and highlights the importance of veterinary surveillance in anticipating the emergence of diseases among humans. The detection of all pathogens, with the exception of Babesia spp. in questing ticks, underlines the need of raising awareness among public and professionals on other tick-borne diseases along with lyme borreliosis.