José A. Oteo

Prevalence of spotted fever group Rickettsia species detected in ticks in La Rioja, Spain.

Abstract:  Our objective was to learn the prevalence of spotted fever group (SFG) Rickettsia detected in ticks in La Rioja, in the north of Spain. From 2001 to 2005, 496 ticks representing 7 tick species were analysed at the Hospital de La Rioja. Ticks were removed from humans with or without rickettsial syndrome (n= 59) or collected from mammals (n= 371) or from vegetation by dragging (n= 66). The presence of SFG Rickettsia in these ticks was investigated by semi‐nested PCR (ompA gene) and sequencing. A phylogenetic tree using Clustal method (neighbor‐joining) was constructed with these data. Only 3 of 170 Hyalomma marginatum ticks carried SFG Rickettsia. Sequencing analysis demonstrated the presence of Rickettsia aeschlimannii (1.8%). Furthermore, Rickettsia massiliae and BAR29 were found in 3 of 120 Rhipicephalus sanguineus specimens (2.5%). In contrast, 81 of 83 tested Dermacentor marginatus ticks were PCR‐positive (97%). Rickettsia slovaca (40.6%) and Rickettsia sp. strains RpA4, DnS14, DnS28 and JL‐02 (59.3%) were found within this tick species. No SFG Rickettsia was detected using ompA primers when Ixodes ricinus, Rhipicephalus bursa, Rhipicephalus turanicus, Rhipicephalus eversti eversti, Hyalomma detritum scupense and Rhipicephalus sp. were analyzed. We detected 17.5% of ticks associated with different SFG Rickettsia: R. aeschlimannii, R. massiliae, BAR29, R. slovaca and Rickettsia sp. strains RpA4, DnS14, DnS28 and JL‐02. Their presence has to be taken into account since most of them have been recognized as human pathogens.

Study on seasonal activity in dogs and ehrlichial infection in Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) from southern Uruguay

ABSTRACT The seasonal activity on dogs of Rhipicephalus sanguineus in southern Uruguay has been studiedfrom samples collected in veterinary clinics of the country. From a total of 744 specimens, adults(92%) were active on dogs between August and March (peaking in October) being absent betweenApril and July. Immatures represent more than 80% of captures on dogs in summer. This pattern is wellcorrelated with the climate records in the zone. The search for ehrlichial DNA from tick-derivedextracts (36 pools from 180 ticks) was negative in all the cases. While R. sanguineus has beenincriminated as vector of these pathogens, additional studies are required to understand the role ofthis tick in the transmission of ehrlichial organisms. Key words : Rhipicephalus sanguineus , seasonal activity, dogs, absence of ehrlichial DNA,Uruguay. INTRODUCTIONAbout 74 species are currently recognized inthe tick genus Rhipicephalus Koch, 1844 1 . Rhipicephalus sanguineus is probably the mostwidely distributed tick in the World. It is foundcircumglobally approximately between thelatitudes 50oN and 42oS. Its preferences for dogsas main host have facilitated its worldwidedistribution

Prevalence data of Rickettsia slovaca and other SFG rickettsiae species in Dermacentor marginatus in the southeastern Iberian Peninsula

Abstract:  In southern Spain, Dermacentor marginatus ticks can be infected with several genospecies of spotted fever Group (SFG) Rickettsia. We developed a nested polymerase chain reaction assay by using a species‐specific probe targeting the ompA gene to detect and differentiate between the two groups of rickettsiae previously described in D. marginatus. SFG rickettsia has been detected in 85.15% of ticks studied (26.7% of positives have been to R. slovaca, the causative agent of TIBOLA‐DEBONEL, and 73.3% to SFG rickettsia closely related to strains RpA4–JL‐02‐DnS14–DnS28).

Detection of a non-pathogenic variant of Anaplasma phagocytophilum in Ixodes ricinus from La Rioja, Spain

Abstract: 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.

Effect of Antibiotic Treatment in Patients with DEBONEL/TIBOLA

Abstract: 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.

Which spotted fever group rickettsia are present in Dermacentor marginatus ticks in Spain

The epidemiology of rickettsiae and rickettsial diseases in Spain is still poorly understood. The main sources of knowledge are various seroepidemiology studies. Ideally, these studies should be expanded with the identification (and isolation if necessary) of the rickettsiae. For large-scale tick study, molecular techniques (PCR for detection and PCR-RFLP or direct sequencing for specific identification) are the most efficient methods. Spotted fever group (SFG) Rickettsia present in D. marginatus (Schulze) collected during 1999 − 2001 in Jaén and La Rioja (Spain) were identified using molecular techniques. DNA was extracted using NucleoSpin (Macherey-Nagel) DNA tissue kit from individually processed males and females ticks. The total number of studied ticks were 24 (12 males and 12 females). PCR amplification and primers for citrate synthase ( glta ), 190 kD surface antigen ( ompA ), and 120 kD surface antigen ( ompB ) were used as previously described. 1 Sequencing was made with a CEQ 2000 automatic sequence system (Beckman Coulter) using capillary electrophoresis and a dye terminator cycle sequence protocol. The glta (382 bp), ompA (510 bp), and ompB (532 bp) fragments were amplified from D. marginatus ticks from Jaén and La Rioja regions (Spain). Restriction patterns when ompA fragments were digested with Pst l or Rsa l and when ompB fragments were digested with Rsa l indicate that two rickettsiae were present in the studied samples: R. slovaca and Rickettsia sp. In the case of Rickettsia sp., both ompA and ompB sequences obtained from PCR products were different from those

Guidelines for the Detection of Rickettsia spp

The genus Rickettsia (Rickettsiales: Rickettsiaceae) includes Gram-negative, small, obligate intracellular, nonmotile, pleomorphic coccobacilli bacteria transmitted by arthropods. Some of them cause human and probably also animal disease (life threatening in some patients). In these guidelines, we give clinical practice advices (microscopy, serology, molecular tools, and culture) for the microbiological study of these microorganisms in clinical samples. Since in our environment rickettsioses are mainly transmitted by ticks, practical information for the identification of these arthropods and for the study of Rickettsia infections in ticks has also been added.

Molecular analysis of Crimean-Congo hemorrhagic fever virus and Rickettsia in Hyalomma marginatum ticks removed from patients (Spain) and birds (Spain and Morocco), 2009-2015.

Crimean-Congo hemorrhagic fever virus (CCHFV) was detected in Spain in 2010. The presence of CCHFV in Hyalomma marginatum ticks from migratory birds passing through Morocco during the spring migration strengthened the hypothesis of the arrival of infected ticks transported by birds to the Iberian Peninsula. Furthermore, Hyalomma species are vectors of bacterial infections such as spotted fever rickettsioses. CCHFV and Rickettsia were screened in Hyalomma ticks from Spain attached to patients (n=12) and birds (n=149). In addition, Rickettsia was investigated in 52 Hyalomma ticks from Morocco (previously reported as CCHFV-infected). No sample collected in Spain showed an infection with CCHFV. Two ticks removed from patients (16.7%), as well as 47 (31.5%) and 4 (7.7%) from birds, collected in Spain and Morocco respectively, were infected with Rickettsia aeschlimannii. Rickettsia sibirica subsp. mongolitimonae was also found in 2 ticks from birds collected in Spain (1.3%). The risk of CCHFV-infected ticks attached to migratory birds to reach the North of Spain is low. This study corroborates the presence of R. aeschlimannii in Spain and Morocco, and supports that H. marginatum can be a potential vector of R. sibirica subsp. mongolitimonae in the Iberian Peninsula.

Serologic study of Rickettsia typhi infection among the human population of southern Spain

Endemic or murine typhus (MT) and several spotted fever rickettsioses (SFR) are endemic diseases in the Iberian peninsula. Murine typhus, an acute febrile illness caused by Rickettsia typhi, has been documented in diverse geographic areas, including the Mediterranean, Africa, Southeast Asia, and the United States [1]. In this work, we evaluate the prevalence of past infection due

Utility of a commercial immunoblot kit (BAG-Borrelia blot) in the diagnosis of the preliminary stages of Lyme disease

The aim of this study was to evaluate the usefulness of a commercial immunoblot (IgG and IgM BAG-Borrelia blot) in the serologic diagnosis of the early stages of Lyme disease. A total of 42 sera from patients with Lyme disease (24 patients with localized early stage (LES) and 18 patients with disseminated early stage (DES)) and 129 sera from patients with non-Lyme diseases (specificity control sera) were studied. IgG anti-p41 from Borrelia burgdorferi s.l. was present in 95.2% of patients followed by anti-p41/I PBi (16.7%), anti-p100 (9.5%) and anti-OspA (9.5%). IgM anti-p41 was present in 66.7% of patients, p41/iPBi (54.8%) and OspC (33.3%). IgM against p100, OspA and OspC were more frequent in DES patients (16.7%, 27.8% and 44.4%) than in LES patients (0.0%, 4.2% and 25.0%). In 4.8% of the cases no IgG bands were present and in 26.2% no IgM bands were present. With the exception of isolated p41 bands (59.5%), no band pattern exceeded 17%. Using manufacturers instructions, test sensitivity in diagnosis of the early stage of Lyme disease is 61.9%, specificity 98.4% and positive and negative predictive values 92.8% and 88.8% respectively. Applying the EUCALB 5, 6 or 7 rules sensitivity increased to 73.8% although specificity decreased to 89.9%. Of the 129 specific control sera, 41.8% presented IgG anti-p41 and 10.8% IgM anti-p41. Patients with non-Lyme diseases that presented more IgG and IgM bands were those patients with syphilis (88.2%), patients with anti-HIV antibodies (57.8%) and patients with anti-nuclear antibodies (ANA) (52.3%).