Risk factors affecting the feeding site predilection of ticks on cattle in Ghana.

Ticks are important disease vectors affecting animal health and causing substantial economic loss, especially in the tropics and subtropics. To examine the tick burden of cattle and associated risk factors for tick infestation, ticks were collected from 388 cattle within five regions in Ghana. Most of the cattle were males (50.3%) and generally older than 3 years (65%). Of the animals sampled, 2187 ticks were collected with a mean tick burden of 5.6 ticks per cattle, and the average tick burden on the udder/scrotum being significantly higher than in the anal region (Generalized Linear Mix Model [GLMM], p = 0.01197). The tick species identified were predominantly Amblyomma variegatum (42.6%) and Hyalomma rufipes (26.2%). High proportions of cattle examined were found to have A. variegatum infesting the udder/scrotum. Furthermore, H. rufipes infested mostly the anal region compared to other examined body parts (OR 14.8, 95% CI 8.6-25.4, p < 0.001). Using the GLMM, tick abundance was found to be significantly higher in cattle older than 3 years. The tick burden in the udder/scrotum was higher than that from the chest and leg/thigh of the cattle (GLMM, p < 0.05). The tick burden at the anal region was also significantly higher than the leg/thigh and chest. This study indicates that the preferred attachment sites of ticks on cattle are species-dependent and effective treatment with acaricides should take into consideration the udder/scrotum and anal regions as well as prioritizing older cattle.

Rickettsia africae and other unclassified Rickettsia species of the spotted fever group in ticks of the Western Ghats, India.

The spotted fever group (SFG) of Rickettsia are zoonotic disease-causing pathogens, commonly transmitted by hard ticks to a wide range of hosts, including humans. Rickettsia conorii is the common SFG recognised in India, whereas most of the infections due to other group species go undifferentiated at the species level. Hence, this study was conducted to screen host-seeking ticks in the Western Ghats region, India, for the DNA of SFG Rickettsia. The ticks were collected from Kerala, Goa, and Maharashtra states of India during a survey conducted between November 2017 and January 2018. In total, 288 tick pools were screened for Rickettsia spp. DNA using pan-Rickettsia real-time PCR, and conventional PCR targeting the gltA, OmpA and 17-kDa protein-coding genes. Nucleotide sequences were subjected to phylogenetic analysis using the NCBI BLAST tool to identify submitted sequences with higher homology. Neighbour-joining trees were constructed using the reference sequences of the GenBank database. Overall, Rickettsia spp. DNA was detected in 27.2% (62/228 pools) of host-seeking ticks across the Western Ghats region, with an estimated minimum infection rate of 0.057. Upon phylogenetic analysis, it was identified that the detected sequences were highly similar (> 99% sequence homology) to R. africae, Candidatus R. laoensis and an un-categorised Rickettsia species, and they were widely carried by Haemaphysalis ticks. The current study is the first report of R. africae and Candidatus R. laoensis in ticks in India. Although the pathogenicity of these species is not well documented, they may pose a potential threat to both animal and the human population in this geographical region.

Rickettsia africae and Rickettsia massiliae in ixodid ticks infesting small ruminants in agro-pastoral settlements in Plateau State, Nigeria.

Arthropods, especially ixodid ticks, have been incriminated in the epidemiology of Spotted Fever Group rickettsioses globally leading to an increasing spectrum of emerging and re-emerging zoonoses with attendant consequences on trade and tourism. The objective of this study was to determine the role of ixodid ticks infesting small ruminants in Plateau State, Nigeria, in the epidemiology of Spotted Fever Group Rickettsiae (SFGR) in the study area. DNA from 130 out of 323 ixodid ticks collected from 179 goats and 121 sheep owned by agro-pastoralists in Plateau State were screened for the evidence of SFGR by molecular methods. Six tick species from four genera were identified: Amblyomma, Hyalomma, Rhipicephalus (Boophilus) and Rhipicephalus. Rhipicephalus sanguineus sensu lato (s.l.) was the predominant (54.5%) species among collected ticks. Tick infestation was significantly associated with the species of small ruminants, the sex of the animals and the sampling locations except for Jos South. Conventional PCR targeting the 381 bp of the citrate synthase (gltA) and 820 bp of the outer membrane protein B (ompB) genes detected DNA of SFGR in nine and eight samples, respectively. Sequence analysis revealed that five sequences obtained from Amblyomma variegatum were 99-100% identical to Rickettsia africae and three sequences from Rh. sanguineus (s.l.) were 100% identical to Rickettsia massiliae reported from Spain. To our knowledge, this is the first report of the detection of R. africae DNA in Am. variegatum collected from small ruminants in Plateau State. Ixodid ticks infesting small ruminants in Plateau state harbor DNA of SFGR with potential veterinary and public health implications.

Infestation rates, seasonal distribution, and genetic diversity of ixodid ticks from livestock of various origins in two markets of Yaoundé, Cameroon.

Little is known about the impact of ticks on livestock and humans in Cameroon. This study aimed to determine the prevalence, seasonal variation, and genetic diversity of hard ticks in the country. Ticks were collected during a cross-sectional survey on domestic livestock in two markets of Yaoundé in 2019 and 2020 and identified using morphological keys, 16S ribosomal DNA, (16S rDNA), and the cytochrome c oxidase subunit 1 (Cox1) genes. The infestation rates were 39.18%, 11.53%, and 2.74% in cattle, sheep, and goats respectively. Three genera of ticks were identified, Rhipicephalus, Amblyomma, and Hyalomma comprising eleven tick species. The main species were Rhipicephalus decoloratus (30.25%), R. microplus (24.43%), and Amblyomma variegatum (12.96%). Rhipicephalus spp. (81.31%) and Amblyomma variegatum (51.54%) were abundant during the rainy season, while Hyalomma spp. (83.86%) during the dry season (p-value <0.00001). Cox1 and 16S rDNA analysis showed a high level of genetic diversity among tick species with sequences close to those observed across Africa. Phylogenetic analysis revealed that our R. microplus belong to clade A and we identified R. sanguineus s.l. as R. linnea. This study shows a high tick infestation rate in cattle, while low in small ruminants with an extensive diversity of tick species, including several known vectors of important tick-borne diseases.

Rickettsial infection in ticks from a natural area of Atlantic Forest biome in southern Brazil.

From June 2013 to January 2014, blood sera samples and ticks were collected from domestic dogs and wild small mammals, and ticks from the vegetation in a preservation area of the Atlantic Forest biome (Turvo State Park), and the rural area surrounding the Park in Derrubadas municipality, state of Rio Grande do Sul, southern Brazil. Dogs were infested by Amblyomma ovale and Amblyomma aureolatum adult ticks, whereas small mammals were infested by immature stages of A. ovale, Amblyomma yucumense, Amblyomma brasiliense, Ixodes loricatus, and adults of I. loricatus. Ticks collected on vegetation were A. brasiliense, A. ovale, A. yucumense, Amblyomma incisum, and Haemaphysalis juxtakochi. Three Rickettsia species were molecularly detected in ticks: Rickettsia bellii in I. loricatus (also isolated through cell culture inoculation), Rickettsia amblyommatis in A. brasiliense, and Rickettsia rhipicephali in A. yucumense. The latter two are tick-rickettsia associations reported for the first time. Seroreactivity to Rickettsia antigens were detected in 33.5% (55/164) small mammals and 8.3% (3/36) canine sera. The present study reveals a richness of ticks and associated-rickettsiae in the largest Atlantic Forest Reserve of the state of Rio Grande do Sul, which is characterized by a rich fauna of wild mammals, typical of more preserved areas of this biome. Noteworthy, none of the detected Rickettsia species have been associated to human or animal diseases. This result contrasts to other areas of this biome in Brazil, which are endemic for tick-borne spotted fever caused by Rickettsia rickettsii or Rickettsia parkeri.

Molecular identification of Theileria spp. in ruminants and ticks from southern littoral of Caspian Sea, Iran.

The present study aimed at evaluating the presence of tick-borne apicomplexan parasites including Theileria ovis, Theileria lestoquardi, Theileria annulata, and Theileria orientalis in 92 cattle and 105 sheep from 6 different districts of Guilan and Mazandaran Provinces, in the southern littoral of Caspian Sea. Furthermore, ixodid ticks were collected from the same animals. Stained blood smears were microscopically evaluated for the presence of blood parasites, and a specific PCR was applied for the detection of Theileria species. Besides, ticks were subsequently examined by species-specific PCR. Microscopic examination of blood smears demonstrated no evidence of intraerythrocytic piroplasms. Species-specific diagnostic PCRs demonstrated that 52.17% of sheep blood samples were positive for T. ovis. In addition, 31.03% and 24.13% of cattle blood samples were positive for T. annulata and T. orientalis, respectively. Moreover, 3 species of the ixodid ticks, namely, Rhipicephalus annulatus (58.47%), Ixodes ricinus (29.82%), and Haemaphysalis inermis (11.69%), were identified in Guilan Province, while Hyalomma detritum (73.03%) and Rhipicephalus sanguineus (26.92%) were found in Mazandaran Province. Additionally, by obtaining the data with respect to tick-borne apicomplexan parasites in 122 infected ticks, 35.24%, 22.95%, and 2.45% of tick samples were positive for T. annulata, T. orientalis, and T. ovis, respectively. Species-specific PCR revealed that H. inermis and R. annulatus were positive for T. orientalis. In addition, T. annulata was found in R. annulatus, H. inermis, and H. detritum. Besides, T. ovis was the only species of Theileria found in R. sanguineus. In conclusion, the results revealed that T. annulata infection was prevalent among cattle and ovine theileriosis caused by T. ovis was the only Theileria species found in sheep in the studied areas of the southern littoral of Caspian Sea. R. annulatus, H. inermis, and H. detritum were the main vectors for T. annulata, followed by H. inermis and R. annulatus for T. orientalis, and R. sanguineus for T. ovis.

Prevalence of causative agents of transmissive tick-borned ricketsious in the Crimean peninsula.

The paper presents the results of a study of the prevalence of Ixodid ticks - potential carriers of tick-borne rickettsiosis pathogens. Ectoparasites were collected in various natural and climatic zones of the Crimean Peninsula within the year 2016-2018. As a result of screening with the help of real-time PCR analysis (PCR-RT), a genetic marker (a section of the gltA gene) of the rickettsia group of tick-borne spotted fever was detected in ticks. The most common DNA marker of rickettsia was found in ticks in the eastern regions of the steppe zone - 50,6 %, in the north-western part of the steppe zone this value was 12,0 %. The least amount of rickettsia target DNA was detected in ticks collected in the mountain forest and south bank zones - 4,5 %. As a result of sequencing of positive DNA samples from fragments of the gltA, ompA, ompB, and sca4 genes, the species composition of rickettsias was established. The DNA of 8 species of rickettsia was identified: Circulation of three R. conorii, R. massiliae, R. sibirica subsp. mongolotimonae, R. slovaca, R. aeschlimannii, R. monacensis, R. helvetica, R. raoultii. R. massiliae, R. slovaca, and R. helvetica were established in the Crimean Peninsula for the first time. The peculiarities of the geographical distribution of the identified rickettsia species were determined, which was due to the spread of mites-carriers of pathogens. The revealed diversity of rickettsia species and their vectors, due to the isolation of the areas of the main feeding animals and the established routes of migratory birds, suggests the circulation of other rickettsia species on the territory of the Crimean Peninsula. The obtained results suggest that the diseases of tick-borne rickettsiosis in the Crimean Peninsula can be caused not only by R. conorii, as previously thought, but also by other types of rickettsii.

Analysis of effectivity of the tickborne encephalitis virus detection methods in ixodid ticks.

Tick-borne encephalitis (TBE) is transmissible viral disease widely common in temperate zone of Eurasia. ELISA and PCR are used for express identification of the vector's infection, but the results of the two methods often do not agree. Aim of the work is comparative analysis for TBE virus of Ixodid ticks from nature using complex of methods, including ELISA, PCR, and isolation of the virus in laboratory mice. 18608 Ixodid ticks were collected during 2013-2019 in TBE natural foci of the Baikal Region. The ticks suspensions were examined individually, using ELISA (n=17610) and PCR (n=2999). Suckling mice were inoculated with the suspensions positive in the both tests. The TBEV antigen was found in 1.2 % of ticks in average. All ticks positive in ELISA were examined in PCR (Group 1). Randomly selected part of negative-ELISA samples were examined in PCR too (Group 2). The PCR results were positive in 68.9±3.13 % of the Group 1, with average Ct index 24.6±0.38. Positive results of PCR in Group 2 accounted for just 2.7±0.31 % with average Ct index 31.0±0.70. The average Ct margin of the Groups 1 and 2 is statistically significant (p < 0.001; df = 118). Isolation of strains was significantly more successful in Group 1 (21.7±2.77 %), than in Group 2 (8.2±5.26 %; p < 0.05; df = 50). ELISA is more useful for examining large amounts of ticks. To get a more complex picture about epidemically dangerous part of the vectors in TBE natural foci, the results of the two express-methods is better to sum. The isolation of the virus is useful to carry out of the samples positive in ELISA and PCR concurrently.

Potential tick vectors for Theileria equi in Israel.

Theileria equi Mehlhorn and Schein, 1998 (Piroplasmida: Babesiidae) is an important tick-borne pathogen of horses that is highly endemic in many parts of the world, including Israel. The present study evaluated the potential roles of five hard tick species [Hyalomma excavatum Koch, 1844; Hyalomma marginatum Koch, 1844; Rhipicephalus turanicus Pomerantsev 1936; Rhipicephalus annulatus Say, 1821; Haemaphysalis parva (Neumann, 1897) (all: Ixodida: Ixodidae)], previously found to infest horses in Israel, in acting as vectors for piroplasmosis. For this, DNA was extracted from whole ticks and, when possible, from the salivary glands in each species (n = 10-59). Polymerase chain reaction amplification and sequencing of the 18S rRNA gene were used to detect T. equi in 48 of the 127 ticks (37.8%) and in 21 of the 90 extracted salivary glands (23.3%) in all five species. All but two sequences were classified as T. equi genotype A; the remaining two were classified as genotype D. The findings of this study point to Ha. parva and R. annulatus as potential novel vectors of T. equi, and suggest that parasite genotype selection occurs within the tick vector.

Morphological and molecular identification of ixodid tick species (Acari: Ixodidae) infesting cattle in Uganda.

In Uganda, the role of ticks in zoonotic disease transmission is not well described, partly, due to limited available information on tick diversity. This study aimed to identify the tick species that infest cattle. Between September and November 2017, ticks (n = 4362) were collected from 5 districts across Uganda (Kasese, Hoima, Gulu, Soroti, and Moroto) and identified morphologically at Uganda Virus Research Institute. Morphological and genetic validation was performed in Germany on representative identified specimens and on all unidentified ticks. Ticks were belonging to 15 species: 8 Rhipicephalus species (Rhipicephalus appendiculatus, Rhipicephalus evertsi evertsi, Rhipicephalus microplus, Rhipicephalus decoloratus, Rhipicephalus afranicus, Rhipicephalus pulchellus, Rhipicephalus simus, and Rhipicephalus sanguineus tropical lineage); 5 Amblyomma species (Amblyomma lepidum, Amblyomma variegatum, Amblyomma cohaerens, Amblyomma gemma, and Amblyomma paulopunctatum); and 2 Hyalomma species (Hyalomma rufipes and Hyalomma truncatum). The most common species were R. appendiculatus (51.8%), A. lepidum (21.0%), A. variegatum (14.3%), R. evertsi evertsi (8.2%), and R. decoloratus (2.4%). R. afranicus is a new species recently described in South Africa and we report its presence in Uganda for the first time. The sequences of R. afranicus were 2.4% divergent from those obtained in Southern Africa. We confirm the presence of the invasive R. microplus in two districts (Soroti and Gulu). Species diversity was highest in Moroto district (p = 0.004) and geographical predominance by specific ticks was observed (p = 0.001). The study expands the knowledge on tick fauna in Uganda and demonstrates that multiple tick species with potential to transmit several tick-borne diseases including zoonotic pathogens are infesting cattle.

Identification of tick-borne pathogens in ticks collected from wild animals in Turkey.

Many enzootic life cycles involving wild animals and non-nidicolous ixodids are still unknown. Therefore, the aim of this study was to report the identified tick species collected from seven different animal species (red deer, brown bear, gray wolf, Eurasian lynx, red fox, European hare, and Mediterranean spur-thighed tortoise) living in the wild in Turkey and to investigate the presence of a wide range of tick-borne microorganisms in the tick samples obtained from these animals. The collected ticks (n = 98) were identified as Dermacentor reticulatus, Haemaphysalis parva, Hyalomma aegyptium, Hyalomma excavatum, Hyalomma marginatum, Ixodes ricinus, and Rhipicephalus turanicus. All engorged ticks collected from the wild animals and unfed larvae (n = 30) obtained from a single Rh. turanicus female were also analyzed individually for tick-borne bacterial and protozoan agents via PCR-sequencing. The molecular analyses revealed the presence of Babesia sp. tavsan2, Theileria capreoli, four Hepatozoon spp. (Hep. ursi, Hep. canis, Hep. felis, and Hepatozoon sp.), Hemolivia mauritanica, and three SFG rickettsiae (Candidatus Rickettsia barbariae, Ca. R. goldwasserii, and Rickettsia hoogstraalii) in the collected ticks. This represents the first report of Th. capreoli, Hep. ursi, and Ca. R. barbariae in ticks from Turkey. The evolutionary relationships of microbes in the different host and tick species are also discussed. Multiple novel tick-host associations in the tick life cycle were also revealed.

Verification of mono- and mixed contaminations transmitting agents tick-borne infections.

In the territory of the Primorsky region, the nosoareas of causative agents of tick-borne infections often coincide and lead to mixed infection of ticks, which causes the mixed pathology in humans . We investigated ixodid ticks taken from people during the epidemic season of 2019 for the spectrum of pathogens of transmissible infections to determine the degree of their mono- and mixed infection. 651 specimens of ixodid ticks investigated. RNA/DNA pathogens was determined by the method of polymerase chain reaction (PCR), TBEV antigen by ELISA. The presence in ticks of pathogens of the following diseases was detected: Lyme borreliosis (35.3%), tick-borne replasing fever (11.3%), human granulocytic anaplasmosis (2.6%), human monocytic ehrlichiosis (2.4%), tick-borne encephalitis (1.8%), tick-borne ricketsiosis (1.0%). As agents of mono-infections, the virus of TBE, ehrlichia, anaplasma and rickettsia in ticks occurred less frequently than in combination with other pathogens. A high frequency of occurrence B. burgdorferi s.l. in mono-infection detected (58.7%). B. miyamotoi has met evenly at ticks with mono- and mixed infections. Total mixed infection of ticks was 23.1%. Borrelia have been identified in all mixed cases. Variants of double combinations (95.5 % of cases from all mixed infections) were as follows: B. burgdorferi s. l. + A. phagocytophilum, B. burgdorferi s.l. + E. chaffeensis / E. muris, B. burgdorferi s.l. + TBEV, B. burgdorferi s.l. + R. heilongjiangensis, B. burgdorferi s.l. + B. miyamotoi. The most frequent combination was B. burgdorferi s.l. + B. miyamotoi (37.8% of all mixed infections). In triple infection of B. burgdorferi s.l. + A. phagocytophilum + TBEV and B. burgdorferi s.l. + A. phagocytophilum + B. miyamotoi, the frequency of occurrence for each combination was 2.2%. Given the above, a complex laboratory diagnosis of tick-borne infections in individuals who have been bitten by a tick is necessary, including the detection of all pathogens common in focal areas.

Tamdy Virus in Ixodid Ticks Infesting Bactrian Camels, Xinjiang, China, 2018.

We isolated Tamdy virus (TAMV; strain XJ01/TAMV/China/2018) from Hyalomma asiaticum ticks infesting Bactrian camels in Xinjiang, China, in 2018. The genome of the strain showed high nucleotide similarity with previously described TAMV strains from Asia. Our study highlights the potential threat of TAMV to public health in China.

Preliminary investigation of ixodid ticks in Jiangxi Province of Eastern China.

In recent years, a large effort has been made for tick surveys for public health importance around China, especially after outbreaks of severe fever with thrombocytopenia syndrome (SFTS) occurred in humans in 2009. In this paper, the preliminary species composition and population distribution of ticks in Jiangxi Province of Eastern China is reported. Ticks were collected in three habitats (grassland, shrubs and woodland) and from nine host groups in 12 sampling sites throughout Jiangxi Province between 2011 and 2018. Six tick species including Haemaphysalis longicornis, Rhipicephalus sanguineus sensu lato, Haemaphysalis yeni, Haemaphysalis kitaoka, Ixodes sinensis and Dermacentor auratus were collected from the vegetation. Haemaphysalis longicornis was most abundant tick species, accounting for 90.6% of the total ticks. Haemaphysalis yeni and H. kitaoka were newly recorded tick species in Jiangxi Province. Tick presence was remarkably greater in grassland (89.4%) than in woodland (9.4%) and shrubs (1.2%), and nymphs (68.2%) and larvae (19.1%) were more frequently found than adult females (6.6%) and males (6.0%). On hosts, a total of 1513 ticks, from 13 species and four genera, were collected. These were H. longicornis, Haemaphysalis campanulata, Haemaphysalis flava, Haemaphysalis phasiana, H. yeni, H. kitaoka, Haemaphysalis hystricis, R. sanguineus (s.l.), Rhipicephalus haemaphysaloides, Rhipicephalus microplus, Ixodes granulatus, I. sinensis and Amblyomma testudinarium. Amblyomma testudinarium was a newly recorded tick species in Jiangxi Province. Based on this investigation, H. longicornis was the most frequently collected species (30.5%) and widely distributed tick species of the total collection ticks (in 11 sampling sites). Haemaphysalislongicornis had a broad host range and its presence (hosts with at least one tick) was significantly greater on Lepus sinensis (33.3%) than on Canis familiaris (2.3%) (χ2 = 23.68, p = 0.0013). In addition, the number of H. longicornis collected on L. sinensis (64.0%) was higher than on other host groups. Of all ticks collected on hosts, different developmental stages were obtained, which included 347 larvae (22.9%), 249 nymphs (16.5%), 404 adult males (26.7%) and 513 females (33.9%) and sex distribution was relatively uniform. These data indicate that a broad range of tick species is widely distributed throughout Jiangxi Province in Eastern China.

[Molecular-genetic monitoring as the basis of modern epidemiological surveillance of tick-borne infections.]

Molecular genetic monitoring of natural focal of tick-borne infections in the epidemic season of 2018 revealed infectiousness of ixodid ticks causative agents of tick-borne encephalitis (0.58% of cases), Lyme disease (31% of cases), human monocytic ehrlichiosis (1.6% of cases) and granulocytic anaplasmosis (3.9% of cases) is registered and also co-infections of ticks by these infections (2.9% of cases) is revealed in natural and anthropourgic foci (B. burgdorferi s.l.+A. phagocytophilum, B. burgdorferi s.l.+E. chaffeensis/E. muris and B. burgdorferi s.l.+tick-borne encephalitis virus). The major epidemiological importance of ticks of the species I. persulcatus is found, their share being 87,6%. The majority of patients being bitten by a tick were from the southern and southeast areas of Primorye. Contamination of ticks with Borrelia was revealed not only in I. persulcatus, but also in ticks of the Haemaphysalis and Dermacentor. The infectiousness of ticks of B. burgdorferi s.l. (42,3%), tick-borne encephalitis virus (7,7%) and A. phagocytophilum (15,4%) was highest on Russky Island.

Identification of an anellovirus and genomoviruses in ixodid ticks.

Ticks are blood-feeding arachnids that are vectors of several important human and animal pathogens. Little is known about single-stranded DNA (ssDNA) viruses that are associated with these ectoparasites. As part of a pilot study to identify ssDNA viruses present in ticks, female American dog ticks (Dermacentor variabilis) and blacklegged ticks (Ixodes scapularis) were collected in eastern Pennsylvania (USA), and the extracted viral DNA was analyzed using viral metagenomics approaches. Three genomoviruses were recovered from pooled samples of D. variabilis (n = 2) and I. scapularis (n = 1): two belonging to the genus Gemycircularvirus, sharing < 63% pairwise identity with other members within the genus; and the third belonging to the genus Gemykolovirus, sharing < 70% pairwise identity to other gemykoloviruses. Furthermore, a genome of an anellovirus belonging to the sharing 62-65% nucleotide identity with torque teno canis viruses (genus Thetatorquevirus) was also recovered from a D. variabilis pooled sample.

Some aspects on tick species in Mongolia and their potential role in the transmission of equine piroplasms, Anaplasma phagocytophilum and Borrelia burgdorferi L.

Ticks are cosmopolitan vectors of numerous diseases, and detection of various pathogens in ticks can help to assess their distribution. In the current study, 528 adult ticks were collected from grazing animals or the ground in ten different Mongolian provinces. Dermacentor nuttalli constituted 76.1% of them and was found in all ecozones except the eastern desert. Dermacentor marginatus (8.3%), Dermacentor silvarum (1.1%) and Ixodes persulcatus (3.0%) were found in the northern forest areas and Hyalomma asiaticum (11.4%) only in the southern (semi-)desert. Of these, 359 ticks were subjected to DNA extraction and PCR was carried out to detect various pathogens. Anaplasma spp. was found in D. marginatus and D. nuttalli (2.5% positive each), including flagged specimen and identified as Anaplasma phagocytophilum. Borrelia spp. were found in 2.5% of the ticks (mostly in I. persulcatus) and identified as Borrelia garinii. Babesia spp. (40%) identified as Babesia caballi were detected in all five tick species including flagged Dermacentor spp. and I. persulcatus, and 3.5% of the ticks (all species except D. silvarum) were positive for Theileria spp. identified as Theileria equi. The piroplasms were found in all provinces. Tick-borne encephalitis virus was not detected. The results highlight the high risk of equine piroplasmosis in Mongolia, which is a concern for both the nomadic population who rely on horses for transport and for conservation of Przewalski's horses in Mongolia. In addition, zoonotic agents such as the avian B. garinii and A. phagocytophilum were also detected, outlining a high risk for exposed humans.

Ixodid ticks of African buffalo (Syncerus caffer), impala (Aepyceros melampus) and elephant (Loxodonta africana) in five protected park estates in the Zambezi valley, Zimbabwe.

Wildlife hosts many pathogens of economic importance and is considered as a reservoir of important tick-borne diseases of livestock in southern Africa. The species composition of ticks parasitizing buffalo (Syncerus caffer), impala (Aepyceros melampus) and elephant (Loxodonta africana) was investigated in five protected parks in the Zambezi valley, Zimbabwe. A total of 1104 adult ticks was collected from 75 adult animals comprising five buffaloes, five elephants and five impalas drawn from five protected wildlife parks. Five tick species belonging to two genera were recovered, with Rhipicephalus decoloratus being the most prevalent species in all the three animal groups. Amblyomma hebraeum was only recovered from buffaloes whereas Rhipicephalus zambeziensis was recovered from buffalos and elephants. Significant differences in mean tick species distribution and concentration were observed amongst the wildlife parks and these appeared to be influenced by the number of hosts in each park. The study revealed that buffaloes are the major host of R. decoloratus in the Zambezi valley. The presence of these ixodid ticks within the Zambezi valley may have significant ecological and economic impacts on wildlife conservation, domestic animals and human health.

Importance of Common Wall Lizards in the Transmission Dynamics of Tick-Borne Pathogens in the Northern Apennine Mountains, Italy.

During the investigations on ticks and tick-borne pathogens (TBP) range expansion in the Northern Apennines, we captured 107 Podarcis muralis lizards. Sixty-eight animals were infested by immature Ixodes ricinus, Haemaphysalis sulcata and H. punctata. Borrelia burgdorferi s.l. was detected in 3.7% of I. ricinus larvae and 8.0% of nymphs. Together with the species-specific B. lusitaniae, we identified B. garinii, B. afzelii and B. valaisiana. Rickettsia spp. (18.1% larvae, 12.0% nymphs), namely R. monacensis, R. helvetica and R. hoogstraalii, were also found in I. ricinus. R. hoogstraalii was detected in H. sulcata nymphs as well, while the two H. punctata did not harbour any bacteria. One out of 16 lizard tail tissues was positive to R. helvetica. Our results support the hypothesis that lizards are involved in the epidemiological cycles of TBP. The heterogeneity of B. burgdorferi genospecies mirrors previous findings in questing ticks in the area, and their finding in attached I. ricinus larvae suggests that lizards may contribute to the maintenance of different genospecies. The rickettsiae are new findings in the study area, and R. helvetica infection in a tail tissue indicates a systemic infection. R. hoogstraalii is reported for the first time in I. ricinus ticks. Lizards seem to favour the bacterial exchange among different tick species, with possible public health consequences.

Francisella tularensis prevalence and load in Dermacentor reticulatus ticks in an endemic area in Central Europe.

A total of 7778 host-seeking adult Dermacentor reticulatus (Ixodida: Ixodidae) ticks were examined for the prevalence of Francisella tularensis holarctica (Thiotrichales: Francisellaceae) in a natural focus of tularaemia in the floodplain forest-meadow ecosystem along the lower reaches of the Dyje (Thaya) river in South Moravia (Czech Republic) between 1995 and 2013. Ticks were pooled (10 specimens per pool) and their homogenates inoculated subcutaneously in 4-week-old specific pathogen-free mice. Dead mice were sectioned, their spleens cultivated on thioglycollate-glucose-blood agar and impression smears from the spleen, liver and heart blood were Giemsa-stained. Sixty-four pools were positive for F. tularensis: the overall minimum infection rate (MIR) was 0.82%. Overall MIRs for the 4714 female and 3064 male D. reticulatus examined were 0.89 and 0.72%, respectively; MIRs fluctuated across years between 0.0 and 2.43%. The estimated bacterial load in infected ticks varied from 0.84 to 5.34 log10 infectious F. tularensis cells per tick (i.e. from about seven to 220 000 cells). Ticks with low loads were more prevalent; more than 1000 infectious cells were detected in 24 ticks (0.3% of all ticks and 37.5% of infected ticks). Monitoring of D. reticulatus for the presence and cell numbers of F. tularensis may be a valuable tool in the surveillance of tularaemia.