Theileria ovis (Piroplasmida: Theileriidae) Detected in Melophagus ovinus (Diptera: Hippoboscoidea) and Ornithodoros lahorensis (Ixodida: Argasidae) Removed From Sheep in Xinjiang, China.

Theileria spp. are tick-transmitted, intracellular apicomplexan protozoan parasites that infect a wide range of animals and, as such, can cause significant economic losses. The aim of the present study was to detect and analyze apicomplexan parasites from two different ectoparasites that were collected from Xinjiang Uygur Autonomous Region, China. The PCR-based detection of 18S rRNA indicated that Ornithodoros lahorensis specimens from Kashgar, Xinjiang, and Aksu were positive for Theileria spp., as were Melophagus ovinus specimens from Aksu. Meanwhile, phylogenetic analysis, based on the 18S rRNA gene sequences, revealed that the four amplified Theileria sequences could be attributed to T. ovis. To the best of our knowledge, this is the first study to report the detection of T. ovis DNA in M. ovinus and the first molecular identification study to confirm the detection of T. ovis in O. lahorensis in China. Accordingly, the present study extends the known distribution of T. ovis.

Detected microorganisms and new geographic records of Ornithodoros rietcorreai (Acari: Argasidae) from northern Brazil.

Reliable data on distributional ranges of soft ticks (Argasidae) and assessments of putative tick-borne agents enhance the understanding on tick-associated microorganisms. A total of 96 ticks morphologicaly and molecularly identified as Ornithodoros rietcorreai were collected in Tocantins State, Brazil, using Noireau traps with living bait as CO2 source. Ninety-six ticks (54 nymphs, 32 males, 10 females) with different engorgement degrees were collected. Fourty-seven (48.9%) of them were individually screened by PCR for detecting bacteria of Anaplasmataceae family and genera Rickettsia, and Borrelia. The presence of protozoans of the genus Babesia was assessed as well. Fourty seven ticks were submitted to analysis. Nine ticks (19.1%) yielded sequences for gltA and htrA genes most identical with a series of endosymbiont rickettsiae and Rickettsia bellii, respectively. Upon two ticks (4.2%) we retrieved DNA of a potential new Wolbachia sp., and DNA of a putative novel Hepatozoon was characterized from three (6.4%) specimens. No DNA of Babesia or Borrelia was detected. Remarkably, amplicons of unidentified eukaryotic organisms, most closely related with apicomplexans but also with dinoflagellates (91% of identity after BLAST analyses), were recovered from two ticks (4.2%) using primers designed for Babesia 18S rRNA gene. Our records expand the distribution of O. rietcorreai into Brazilian Cerrado biome and introduce the occurrence of microorganisms in this tick species.

Shifts in Borrelia burgdorferi (s.l.) geno-species infections in Ixodes ricinus over a 10-year surveillance period in the city of Hanover (Germany) and Borrelia miyamotoi-specific Reverse Line Blot detection.

BACKGROUND: Lyme borreliosis caused by spirochetes of the Borrelia burgdorferi (sensu lato) complex is still the most common tick-borne disease in Europe, posing a considerable threat to public health. The predominant vector in Europe is the widespread hard tick Ixodes ricinus, which also transmits the relapsing fever spirochete B. miyamotoi as well as pathogenic Rickettsiales (Anaplasma phagocytophilum, Rickettsia spp.). To assess the public health risk, a long-term monitoring of tick infection rates with the named pathogens is indispensable. METHODS: The present study is the first German 10-year follow-up monitoring of tick infections with Borrelia spp. and co-infections with Rickettsiales. Furthermore, a specific Reverse Line Blot (RLB) protocol for detection of B. miyamotoi and simultaneous differentiation of B. burgdorferi (s.l.) geno-species was established. RESULTS: Overall, 24.0% (505/2100) of ticks collected in the city of Hanover were infected with Borrelia. In detail, 35.4% (203/573) of adult ticks [38.5% females (111/288) and 32.3% males (92/285)] and 19.8% nymphs (302/1527) were infected, representing consistent infection rates over the 10-year monitoring period. Geno-species differentiation using RLB determined B. miyamotoi in 8.9% (45/505) of positive ticks. Furthermore, a significant decrease in B. afzelii and B. spielmanii infection rates from 2010 to 2015 was observed. Co-infections with Rickettsia spp. and A. phagocytophilum increased between 2010 and 2015 (7.3 vs 10.9% and 0.3 vs 1.1%, respectively). CONCLUSIONS: Long-term monitoring is an essential part of public health risk assessment to capture data on pathogen occurrence over time. Such data will reveal shifts in pathogen geno-species distribution and help to answer the question whether or not climate change influences tick-borne pathogens.

Conspecific hyperparasitism: An alternative route for Borrelia hermsii transmission by the tick Ornithodoros hermsi.

Ixodid and argasid ticks may hyperparasitize other individuals of their own species to acquire a blood meal, however most accounts are based on single observations and the behavior has rarely been studied. While maintaining laboratory colonies of Ornithodoros species, we noticed that unfed ticks occasionally fed on other ticks that were feeding on mice, and unfed ticks parasitized engorged ticks when confined together in tubes. Therefore, we investigated hyperparasitism by Ornithodoros hermsi and the ability of these ticks to acquire and transmit the relapsing fever spirochete Borrelia hermsii when feeding on other ticks. Various combinations of unfed and recently engorged male, female and nymphal ticks were confined for 1-2h as individual pairs or in groups, then examined to determine the number of ticks that acquired blood by feeding on others. Unfed O. hermsi males were far more likely to hyperparasitize other ticks than were females and nymphs, as 78.6% of males (114 of 145 ticks) fed when confined with recently engorged ticks. Unfed females and nymphs also hyperparasitized other ticks but far less frequently (only 6.7% combined; 17 of 254 ticks). Infection experiments demonstrated that unfed males acquired B. hermsii when parasitizing nymphs that had recently engorged on a spirochetemic mouse, and unfed infected males transmitted spirochetes to recently engorged nymphs. Some ticks infected via hyperparasitism subsequently transmitted B. hermsii to mice. Hyperparasitism by O. hermsi occurred more frequently than expected, although possibly influenced by our experimental design. The significance of this behavior as it may influence the horizontal transfer of B. hermsii in nature is not known but worthy of future consideration.

Detection of relapsing fever Borrelia spp., Bartonella spp. and Anaplasmataceae bacteria in argasid ticks in Algeria.

BACKGROUND: Argasid ticks (soft ticks) are blood-feeding arthropods that can parasitize rodents, birds, humans, livestock and companion animals. Ticks of the Ornithodoros genus are known to be vectors of relapsing fever borreliosis in humans. In Algeria, little is known about relapsing fever borreliosis and other bacterial pathogens transmitted by argasid ticks. METHODOLOGY/PRINCIPAL FINDINGS: Between May 2013 and October 2015, we investigated the presence of soft ticks in 20 rodent burrows, 10 yellow-legged gull (Larus michahellis) nests and animal shelters in six locations in two different bioclimatic zones in Algeria. Six species of argasid ticks were identified morphologically and through 16S rRNA gene sequencing. The presence and prevalence of Borrelia spp., Bartonella spp., Rickettsia spp. and Anaplasmataceae was assessed by qPCR template assays in each specimen. All qPCR-positive samples were confirmed by standard PCR, followed by sequencing the amplified fragments. Two Borrelia species were identified: Borrelia hispanica in Ornithodoros occidentalis in Mostaganem, and Borrelia cf. turicatae in Carios capensis in Algiers. One new Bartonella genotype and one new Anaplasmataceae genotype were also identified in Argas persicus. CONCLUSIONS: The present study highlights the presence of relapsing fever borreliosis agents, although this disease is rarely diagnosed in Algeria. Other bacteria of unknown pathogenicity detected in argasid ticks which may bite humans deserve further investigation.

Novel Babesia and Hepatozoon agents infecting non-volant small mammals in the Brazilian Pantanal, with the first record of the tick Ornithodoros guaporensis in Brazil.

Taking into account the diversity of small terrestrial mammals of the Pantanal, the present study aimed to verify the occurrence of infection by Ehrlichia spp., Anaplasma spp., Rickettsia spp., Hepatozoon spp., Babesia spp. and parasitism by ticks in non-volant small mammals collected in the Brazilian Pantanal. Samples of blood, liver and spleen were collected from 64 captured animals, 22 marsupials and 42 rodents. Pathogen detection was performed by the use of genus-specific Polymerase Chain Reaction (PCR) assays. Ticks collected from the animals consisted of Amblyomma sculptum and Amblyomma triste nymphs, and Ornithodoros guaporensis larvae. None of the vertebrate samples (blood, liver, or spleen) yielded detectable DNA of Rickettsia spp. or Ehrlichia spp. The blood of the rodent Hylaeamys megacephalus yielded an Anaplasma sp. genotype (partial 16S rRNA gene) 99% similar to multiple Anaplasma spp. genotypes around the world. The blood of three rodents of the species Calomys callosus were positive for a novel Hepatozoon sp. agent, phylogenetically related (18S rDNA gene) to distinct Hepatozoon genotypes that have been detected in rodents from different parts of the world. One marsupial (Monodelphis domestica) and three rodents (Thrichomys pachyurus) were positive to novel piroplasmid genotypes, phylogenetically (18S rDNA gene) related to Theileria bicornis, Cytauxzoon manul, and Cytauxzoon felis. The present study provides the first molecular detection of Hepatozoon sp. and piroplasmids in small mammals in Brazil. Additionally, we expanded the distribution of O. guaporensis to Brazil, since this tick species was previously known to occur only in Bolivia.

Diversity and distribution of Borrelia hermsii.

Borrelia hermsii is the most common cause of tickborne relapsing fever in North America. DNA sequences of the 16S-23S rDNA noncoding intergenic spacer (IGS) region were determined for 37 isolates of this spirochete. These sequences distinguished the 2 genomic groups of B. hermsii identified previously with other loci. Multiple IGS genotypes were identified among isolates from an island, which suggested that birds might play a role in dispersing these spirochetes in nature. In support of this theory, all stages of the tick vector Ornithodoros hermsi fed successfully on birds in the laboratory and advanced in their life cycle. B. hermsii produced a detectable spirochetemia in 1 chicken inoculated subcutaneously. Additional work is warranted to explore the role of birds as enzootic hosts for this relapsing fever spirochete.

Babesia parasites develop and are transmitted by the non-vector soft tick Ornithodoros moubata (Acari: Argasidae).

Ornithodoros moubata ticks were fed on blood infected with Babesia equi. However, the parasites were quickly cleared as evidenced by the disappearance of B. equi-specific ribosomal RNA from the ticks. We hypothesized that if the Babesia parasite can escape midgut-associated barriers a non-vector tick can become infected with Babesia. To test this hypothesis, B. equi parasite-infected blood from in vitro culture was injected into the haemocoel of ticks. B. equi-specific rRNA was surprisingly detected 45 days after injection even in the eggs. Babesia-free dogs were infested with O. moubata ticks that were infected by inoculation with B. gibsoni-infected red blood cells. Parasitaemia and antibody production against Bg-TRAP of B. gibsoni increased gradually. These results indicate that O. moubata may be a useful vector model for Babesia parasites and also a very important tool for studies on tick immunity against Babesia parasites and tick-Babesia interactions.