Q fever - from diagnosis to eco-epidemiological investigation of Coxiella burnetii in the context of human infection

Funder

Organizational Unit

Publications

REVIVE, a surveillance program on vectors and vector-borne pathogens in Portugal - four year experience on ticks

Publication . Santos, A.S.; Santos Silva, M.; Lopes de Carvalho, I.; Milhano, N.; Chaínho, L.; Luz, T.; Parreira, P.; Gomes, S.; De Sousa, R.; Núncio, M.S.; REVIVE Workgrup

REVIVE is a national wide surveillance program on vector and vector-borne agents implement and coordinate by the National Institute of Health (CEVDI/INSA) in collaboration with other institutions of the Health Ministry. The programme started in 2008 with the surveillance of mosquitoes and later in 2011 was extended to ticks. The main goals of this project are to collect and identify vectors, updating our knowledge in the distribution, hostassociations, seasonality and abundance of the Portuguese species. Additionally this project contributes for monitoring the introduction of exotic vector species. This work regards the 4-year REVIVE studies on ticks and Borrelia/Rickettsia surveillance, among other tick-borne agents, discussing the established circuits, obtained results and practical interventions. Over 29.000 ticks were collected on hosts or by flagging vegetation from 168 (60.4%) municipalities of mainland Portugal. Collection in humans reached the 583 specimens. In total, 13 autochthonous tick species were identified, including Dermacentor marginatus; D. reticulatus; Haemaphysalis punctata; Hyalomma lusitanicum; H. marginatum; Ixodes canisuga; I. hexagonus; I. ricinus; I. ventalloi; Rhipicephalus annulatus; R. bursa; R. pusillus; R. sanguineus. Of note is the identification of an exotic species, Amblyomma sp., attached to a Portuguese emigrant arriving from USA. The top three species collected during this surveillance program were R. sanguineus (69%), followed by R. pusillus (16.4%) and H. marginatum (9.7%). However regarding antropofilic behaviour, from the 11 species found in humans the most prevalent were I. ricinus (35%), followed by R. sanguineus (34%), and H. marginatum (14%). The abundance, distribution, host association and other relevant patterns are compared with previous existing records. Regarding the tick-borne agents, all ticks collected from humans and about 10% of the questing/host-attached ticks were tested for Borrelia and Rickettsia spp., among other agents. Ten bacteria were identified so far in single or multiple infection, including Borrelia afzelii, B. garinii, B. lusitaniae, Rickettsia aeschlimannii, R. conorii, R. helvetica, R. massiliae, R. monacensis, R. raoulti, and R. slovaca. The importance of including other tick-borne agents in routine screening is also discussed. The presented data reinforces the importance of the REVIVE. The program has contributed to call attention to tick-borne diseases not only among healthcare providers but also in the populations. The workflow established, has also enabled timely screeningof ticks removed from humans, animals or in a given environment, allowing the implementation of informed prevention/control strategies and directly contributing to improve Public Health in Portugal.

2015Conference object

Open access

Coxiella burnetii DNA detected in domestic ruminants and wildlife from Portugal

Publication . Cumbassá, A.; Barahona, M.J.; Cunha, M.V.; Azorín, B.; Fonseca, C.; Rosalino, L.M.; Tilburg, J.; Hagen, F.; Santos, A.S.; Botelho, A.

Coxiella burnetii is the etiological agent of Q fever or Coxiellosis, a zoonosis mainly affecting domestic ruminants. Information on the population structure and epidemiology of C. burnetii in animals is scarce in Portugal. Evidence of C. burnetti infection was sought in domestic, wild and captive animals based on the detection of bacterial DNA. Tissue samples from 152 domestic animals (cattle = 24, goats = 51, sheep = 76 and swine = 1), 55 wild carnivores (Egyptian mongoose = 45, red fox = 4, common genet = 3, weasel = 2 and European badger = 1) and 22 zoo animals (antelopes = 15, impala = 1; rhinoceros = 1, deer = 2, zebras = 2 and giraffe = 1) were screened by nested-touchdown PCR. Cloacae swabs from 19 griffon vultures were also analysed. Among the domestic ruminants, goats presented the highest prevalence of infection (23.53%), followed by cattle, (20.83%) and sheep (10.53%). C. burnetii DNA was also detected in five Egyptian mongooses and two antelopes and one giraffe. Using a 6-locus multiple-locus variable-number tandem repeat analysis (MLVA-6) six complete genotypes, T, I and CM and the first reported CN, CO and CP, were identified, respectively, in small ruminants and Egyptian mongooses. Clustering analysis of genotypes exposed four distinct groups, according to detection source, enlightening an apparent association between C. burnetii genotype and host.

2015-10-22Journal article

Restricted access

Anaplasma marginale and Theileria annulata in questing ticks from Portugal

Publication . Antunes, S.; Ferrolho, J.; Domingues, N.; Santos, A.S.; Santos-Silva, M.M.; Domingos, A.

Ticks are ubiquitous arthropods and vectors of several pathogenic agents in animals and humans. Monitoring questing ticks is of great importance to ascertain the occurrence of pathogens and the potential vector species, offering an insight into the risk of disease transmission in a given area. In this study 428 host-seeking ticks, belonging to nine species of Ixodidae and collected from 17 of the 23 Portuguese mainland subregions, were screened for several tick-borne agents with veterinary relevance: Anaplasma marginale, Anaplasma ovis, Anaplasma centrale, Babesia spp., Coxiella burnetii and Theileria spp. Prevalence was assessed by PCR and amplified amplicons sequenced for validation of results. Twenty ticks, in a total of 428, were found positive: one Ixodes ventalloi for Theileria annulata and four Dermacentor marginatus, one Haemaphysalis punctata, five Ixodes ricinus, five I. ventalloi, and four Rhipicephalus sanguineus sensu lato for A. marginale. According to the reviewed literature, this is the first report of A. marginale and T. annulata detection in I. ventalloi. Furthermore, the amplification of A. marginale DNA in several tick species suggests a broad range for this agent in Portugal that might include other uncommon species as R. sanguineus s.l. This work provides new data towards a better understanding of tick-pathogen associations and also contributes to the surveillance of tick-borne agents in geographic areas with limited information.

2016-07-09Journal article

Restricted access

Detection and phylogenetic characterization of Theileria spp. and Anaplasma marginale in Rhipicephalus bursa in Portugal

Publication . Ferrolho, A.; Antunes, S.; Santos, A.S.; Velez, R.; Padre, L.; Cabezas-Cruz, A.; Santos-Silva, M.M.; Domingos, A.

Ticks are obligatory blood-sucking arthropod (Acari:Ixodida) ectoparasites of domestic and wild animals as well as humans. The incidence of tick-borne diseases is rising worldwide, challenging our approach toward diagnosis, treatment and control options. Rhipicephalus bursa Canestrini and Fanzago, 1877, a two-host tick widely distributed in the Palearctic Mediterranean region, is considered a multi-host tick that can be commonly found on sheep, goats and cattle, and occasionally on horses, dogs, deer and humans. R. bursa is a species involved in the transmission of several tick-borne pathogens with a known impact on animal health and production. The aim of this study was to estimate R. bursa prevalence in Portugal Mainland and circulating pathogens in order to contribute to a better knowledge of the impact of this tick species. Anaplasma marginale and Theileria spp. were detected and classified using phylogenetic analysis. This is the first report of Theileria annulata and Theileria equi detection in R. bursa ticks feeding on cattle and horses, respectively, in Portugal. This study contributes toward the identification of currently circulating pathogens in this tick species as a prerequisite for developing future effective anti-tick control measures.

2016-04Journal article

Restricted access

PCR screening of tick-borne agents in sensitive conservation areas, Southeast Portugal

Publication . Santos-Silva, M.M.; Melo, P.; Santos, N.; Antunes, S.; Duarte, L.R.; Ferrolho, J.; Milhano, N.; Santos, P.T.; Domingos, A.; Santos, A.S.

The Southeast region of Portugal, particularly the Guadiana valley, is currently the reintroduction territory of Lynx pardinus (Iberian lynx), one of the most endangered felids in the world that is only found in the Iberian Peninsula. Over the last century, populations have declined, placing L. pardinus at extremely high risk of extinction in the wild and relying on reintroduction projects. Among the aspects taken into account in the establishment of new populations is the sanitary status of the selected habitats, especially concerning infectious diseases, including tick-borne pathogens (TBPs). This study presents the results of TBPs survey on ticks collected at sensitive conservation areas of Southeast Portugal. From 2012 to 2014, 231 ticks obtained from vegetation, sympatric domestic and wild animals were submitted for analysis. The presence of Babesia spp., Cytauxzoon spp., Theileria spp., Hepatozoon spp., Anaplasma spp., Ehrlichia spp., Candidatus Neoehrlichia mikurensis, among other Anaplasmataceae, and Coxiella burnetii were investigated by PCR. Six tick species were recorded, Dermacentor marginatus (n = 13/5.6%), Hyalomma lusitanicum (n = 175/75.8%), Ixodes ricinus (n = 4/1.7%), Rhipicephalus bursa (n = 7/3.0%), R. pusillus (n = 21/9.1%) and R. sanguineus sensu lato (n = 11/4.8%). The molecular screening confirmed the presence of two tick-borne pathogens, C. burnetii (N = 34) and Anaplasma platys (N = 1), and one tick-endosymbiont, Candidatus Midichloria mitochondrii (N = 45). The results obtained provide new information on the circulation of ticks and TBPs with potential veterinary importance in Iberian lynx habitat.

2016-11Journal article

Restricted access