Aránzazu Portillo

The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes species of medical and veterinary importance. The presence of Anaplasma spp. in ticks from birds, as well as in Haemaphysalis punctata (Ixodida: Ixodidae) specimens collected from cattle and vegetation in northern Spain was investigated. A total of 336 ticks from birds [174 Ixodes frontalis (Ixodida: Ixodidae), 108 H. punctata, 34 Hyalomma marginatum (Ixodida: Ixodidae), 17 Ixodes ricinus (Ixodida: Ixodidae) and three Ixodes spp.], and 181 H. punctata specimens collected from cattle (n = 71) and vegetation (n = 110) were analysed. Anaplasma bovis was detected in five H. punctata, including two from birds (1.9%) and three from vegetation (2.7%). Four I. frontalis (2.3%) (one co-infected with 'Candidatus Midichloria mitochondrii') and one I. ricinus (5.9%) removed from birds, as well as four H. punctata (5.6%) collected from cattle showed Anaplasma phagocytophilum infection. In addition, Anaplasma centrale was found in two H. punctata, one from a cow (1.4%) and the other from vegetation (0.9%). This study represents the first evidence of the presence of A. bovis in European ticks, and reports the first detection of A. bovis and A. centrale in H. punctata, and the first finding of A. phagocytophilum and 'Ca. Midichloria mitochondrii' in I. frontalis.

Rickettsial infections are caused by intracellular bacteria. They do not grow in standard culture media so there are limitations in routine practice to study their metabolism. Nuclear Magnetic Resonance (NMR) spectroscopy is used for identification of metabolites in biological samples. Vero cells infected with Rickettsia slovaca as well as uninfected cells were monitored by (1)H NMR showing the presence of ethanol and lactic acid. As no differences were observed, labeled compounds were added into cultures. When D-[1-13C]glucose was monitored by (13)C NMR no differences among infected and uninfected cells were observed in metabolic profiles. Glucose was transformed into ethanol in all cultures. Monitored experiments carried out with [2-13C]glycine showed differences between infected and uninfected cell cultures spectra. Glycine was partially transformed into serine, but the amount of the serine formed was larger in those infected. Moreover, L-[2-13C]leucine, L-[1-13C]isoleucine and L-[15N]tyrosine were evaluated. No differences among infected and uninfected cells were observed in the metabolic profiles when tyrosine and leucine were monitored. The amino acid L-[1-13C]isoleucine exhibited different metabolism in presence of the R. slovaca, showing a promising behavior as biomarker. In this work we focused on finding one or more compounds that could be metabolized specifically by R. slovaca and could be used as an indicator of its activity.

Ticks are involved in the epidemiology of several human pathogens including spotted fever group (SFG) Rickettsia spp., Coxiella burnetii and Bartonella spp. Human diseases caused by these microorganisms have been reported from South Africa. The presence of SFG Rickettsia spp., C. burnetii and Bartonella spp. was investigated in 205 ticks collected from domestic and wild animals from Western Cape and Limpopo provinces (South Africa). Rickettsia massiliae was detected in 10 Amblyomma sylvaticum and 1 Rhipicephalus simus whereas Rickettsia africae was amplified in 7 Amblyomma hebraeum. Neither C. burnetii nor Bartonella spp. was found in the examined ticks. This study demonstrates the presence of the tick borne pathogen R. massiliae in South Africa (Western Cape and Limpopo provinces), and corroborates the presence of the African tick-bite fever agent (R. africae) in this country (Limpopo province).

Rickettsioses are caused by obligate intracellular bacteria within the genus Rickettsia, mainly transmitted by arthropods. Until recently, Mediterranean spotted fever (MSF) caused by Rickettsia conorii was considered the only tick-borne rickettsiosis in Europe. However, 'new' TBR have been described in Europe during last years. For instance, other subspecies such as R. conorii caspia and R. conorii israelensis have been involved in MSF. Dermacentor-borne necrosis erythema and lymphadenopathy/tick-borne lymphadenopathy (DEBONEL/TIBOLA) cases caused by Rickettsia slovaca, Rickettsia raoultii, and Rickettsia rioja been described in several countries where Dermacentor marginatus ticks (the mainly implicated vector) are present. Rickettsia helvetica has also been involved as a human pathogen in cases of fever with and without rash and in patients with meningitis and carditis. Other TBR such as lymphangitis-associated rickettsioses (LAR), caused by Rickettsia sibirica mongolitimonae, have been diagnosed in different European countries (France, Spain, Portugal and Greece). Rickettsia massiliae is considered an etiological agent of MSF-like illness in the Mediterranean basin. Furthermore, Rickettsia monacensis that is distributed all along Europe has been isolated from patients with MSF-like illness in Spain. Although Rickettsia aeschlimannii has been associated with MSF-like in Africa and is distributed in the Mediterranean area, no autochthonous human cases have been reported for Europe. Other Rickettsia species detected in ticks and unrelated to human disease (Candidatus Rickettsia kotlanii, Candidatus Rickettsia barbariae, Candidatus Rickettsia vini) could be potentially involved in the next years. Climate changes, among other factors, may contribute to the emergence of other rickettsioses or change their distribution. Lastly, African tick-bite fever (ATBF), caused by Rickettsia africae, is frequently diagnosed in Europe in patients returning from endemic areas.

The pathophysiological hallmark of Rickettsia conorii (R. conorii) infection comprises infection of endothelial cells with perivascular infiltration of T-cells and macrophages. Although interferon (IFN)-γ-induced protein 10 (IP-10)/CXCL10 is induced during vascular inflammation, data on CXCL10 in R. conorii infection is scarce. Serum CXCL10 was analyzed in two cohorts of southern European patients with R. conorii infection using multiplex cytokine assays. The mechanism of R. conorii-induced CXCL10 release was examined ex vivo using human whole blood interacting with endothelial cells. (i) At admission, R. conorii infected patients had excessively increased CXCL10 levels, similar in the Italian (n=32, ∼56-fold increase vs controls) and the Spanish cohort (n=38, ∼68-fold increase vs controls), followed by a marked decrease after recovery. The massive CXCL10 increase was selective since it was not accompanied with similar changes in other cytokines. (ii) Heat-inactivated R. conorii induced a marked CXCL10 increase when whole blood and endothelial cells were co-cultured. Even plasma obtained from R. conorii-exposed whole blood induced a marked CXCL10 release from endothelial cells, comparable to the levels found in serum of R. conorii-infected patients. Bacteria alone did not induce CXCL10 production in endothelial cells, macrophages or smooth muscle cells. We show a massive and selective serum CXCL10 response in R. conorii-infected patients, likely reflecting release from infected endothelial cells characterized by infiltrating T cells and monocytes. The CXCL10 response could contribute to T-cell infiltration within the infected organ, but the pathologic consequences of CXCL10 in clinical R. conorii infection remain to be defined.

: DEBONEL/TIBOLA is a tick‐borne acute/sub‐acute infection transmitted in our environment by Dermacentor marginatus and mainly caused by Rickettsia slovaca. The aim of our study was to know the effect of starting early treatment in the course of the DEBONEL/TIBOLA.

Our aim was to identify variants of Anaplasma phagocytophilum 16S rRNA gene sequences among products amplified from Ixodes ricinus collected in La Rioja, Spain. A. phagocytophilum AP-variant 1, reported as non-pathogenic, was detected in 12 samples (two adults and ten nymphs). This finding could justify the low incidence of human anaplasmosis in our area, despite the high prevalence of A. phagocytophilum in ticks.

Lyme disease (LD) is a worldwide-distributed multisystemic process caused by Borrelia burgdorferi sensu lato (s.l.) and transmitted by hard ticks. In fact, it is the most common tick-borne infectious disease in the northern hemisphere. In Spain it is transmitted by Ixodes ricinus ticks and Borrelia garinii is the genoespecies of B. burgdorferi s.l. mostly involved in our area. LD is known as "the last great imitator" due to the broad clinical spectrum that may cause. Except in the case of erythema migrans (pathognomonic feature of the disease), the remaining clinical manifestations should be confirmed using microbiological tests. This review is intended to provide readers a current vision of the etiology, epidemiology, clinical manifestations, laboratory diagnosis and treatment of Lyme disease in our environment. Controversial aspects arising from the use of non-validated microbiological tests that are being used without scientific rigor are highlighted.

In an attempt to know the potential risk of human disease after exposure to ticks in La Rioja (North of Spain), the objective of our study was to investigate the presence of Rickettsia species in Haemaphysalis ticks collected in our area. A total of 177 Haemaphysalis spp. belonging to three species (H. punctata, H. sulcata, and H. inermis) were subjected to DNA extraction and tested by polymerase chain reaction (PCR) targeting three rickettsial genes: gltA, ompB, and ompA. Six (3 H. inermis, 2 H. punctata, and 1 H. sulcata) of the 177 specimens were found to be infected (3.4%). The rickettsiae in H. inermis ticks (n = 3) were identified as Rickettsia aeschlimannii by sequencing of the genes coding for gltA, ompB, and ompA. Nucleotide sequences from H. punctata and H. sulcata samples that yielded PCR products (n = 3), showed >99% similarity with sequences of Rickettsia endosymbiont of H. sulcata and 'Candidatus Rickettsia hoogstraalii' for gltA and ompB genes, respectively. Attempts to amplify ompA from these two H. punctata and one H. sulcata failed. This study suggests that H. inermis could be a vector for tick-borne spotted fever caused by R. aeschlimannii in the north of Spain. Further studies on characterization and culture of rickettsial endosymbionts found in Haemaphysalis spp. should be performed.