Isolation and Identification of a Rickettsial Strain Related to Rickettsia massiliae in Greek Ticks

Babalis, T.; Tselentis, Yannis; Roux, V; Psaroulaki, Anna; Raoult, Didier

doi:10.4269/ajtmh.1994.50.365

Cited by 65 publications

(41 citation statements)

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“…Up to now in Greece, R. conorii, along with two nonpathogenic rickettsial species (Rickettsia rhipicephali and a variant of Rickettsia massiliae) has been found only in the Rhipicephalus species complex ticks (2,8). This study represents the first description of a spotted fever group rickettsia in Hyalomma sp.…”

Section: Case Reportmentioning

confidence: 95%

Simultaneous Detection of “ Rickettsia mongolotimonae ” in a Patient and in a Tick in Greece

Psaroulaki

Germanakis²,

Gikas

et al. 2005

J Clin Microbiol

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Rickettsia conorii, a spotted fever group rickettsia which is transmitted by Rhipicephalus sp. complex ticks, was considered until now the only pathogenic rickettsia prevalent in Greece. Here, we report the presence of "Rickettsia mongolotimonae" (proposed name) detected simultaneously in a patient and in a Hyalomma anatolicum excavatum tick, sampled on the patient. CASE REPORTA 76-year-old man, was admitted on 9 December 2002 to the hospital of Sitia (Crete, Greece) with fever (38.2°C), malaise, marked fatigue, lumbar and shoulder myalgias, and knee and hand arthralgias for 7 days prior to admission. He reported no direct contact with domestic animals except that he had been collecting olives (for 6 days) in fields that were visited by goats and sheep. On the day of admission the physician removed an engorged tick from his scrotum, which was morphologically identified as a female belonging to the Hyalomma anatolicum excavatum species. The patient reported that he removed another engorged tick 3 days before the onset of symptoms.On physical examination, a light discrete hyperemic maculopapular skin rash was observed, with elements 3 to 5 mm in diameter, mostly on the trunk and proximal extremities. There was no inoculation eschar at the tick bite sites. We observed, however, a single, large (17 mm in diameter) papule at the epigastrium with a small (1 mm in diameter) central crust that persisted for about 4 weeks and a painless enlarged lymph node of 1 cm in diameter in the right inguinal region. During the next two days a few elements of the rash presented petechias in their centers.Initial laboratory examination demonstrated moderate anemia (hemoglogin, 11.5 g/dl), marked increase in serum urea (178 mg/dl) and creatinine (6 mg/dl), increased levels of lactate dehydrogenase (1,486 IU/liter) triglycerides and uric acid, elevated C-reactive protein (229 mg/liter), and mildly elevated serum concentrations of aspirate aminotransferase (67 IU/liter) and alanine aminotransferase (72 IU/liter). The electrocardiogram revealed T-wave inversions in leads I and aVL while, at the second day of hospitalization, paroxysmal atrial fibrillation emerged. The patient was treated with doxycycline (200 mg/day for 10 days) and rapidly recovered.Serum samples were taken from the patient on days 7 and 15 after the onset. Antibodies to Rickettsia typhi, Coxiella burnetii, Rickettsia conorii, "Rickettsia mongolotimonae" (proposed name), and Rickettsia sibirica were determined by microimmunofluorescence (antigens were kindly provided by D. Raoult). The first serum was negative for all tested antigens. The second sample yielded immunoglobulin G (IgG) titers of 1/256, 1/1,024, and 1/256 and IgM titers of 1/64, 1/256, 1/128 against R. conorii, R. mongolotimonae, and R. sibirica, respectively, but was negative for C. burnetii and R. typhi.DNA was extracted from the removed tick and the wholeblood sample by using the QIAmp tissue kit and QIAmp blood minikit, respectively (QIAGEN GmbH, Hilden, Germany), according to the manufacturer's instruc...

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Section: Case Reportmentioning

confidence: 95%

Simultaneous Detection of “ Rickettsia mongolotimonae ” in a Patient and in a Tick in Greece

Psaroulaki

Germanakis²,

Gikas

et al. 2005

J Clin Microbiol

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“…Amplification, digestion, and electrophoresis were carried out as described previously. [14][15][16] The PCR amplifications always included a negative control that contained no template DNA, and R. conorii DNA (a Moroccan strain, provided by Pr. D. Raoult, Unit of Rickettsiae, Faculty of Medicine, Marseilles, France) was used as positive control.…”

Section: Methodsmentioning

confidence: 99%

Fourteen-year seroepidemiological study of zoonoses in a Greek village.

Αντωνίου¹,

Economou²,

Wang³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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Abstract. A seroepidemiological study carried out in a high-risk village in Crete in 1985-1987 and 1998 showed that although the awareness of the people concerning zoonoses had increased during this period, the situation did not improve: there was a significant increase of the spread of seroprevalence in time and space of Coxiella burnetii, Rickettsia typhi, Brucella sp., and Entamoeba histolytica. Toxoplasma gondii, Rickettsia conorii, Borrelia burgdorferi, Echinococcus granulosus, Leishmania sp., and Fasciola hepatica stayed at the same levels. This first study of Bartonella henselae in Crete showed that 15.9% of the children tested were seropositive. The results indicate that reservoirs and vectors of the pathogens studied are widespread in the environment, and the way of life of the people favors contact with them. Seven of 30 milk samples were positive for Brucella sp. by seminested polymerase chain reaction.

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“…As a positive control, purified DNA from R. conorii (Moroccan strain) was used. The DNA from ticks that were positive for the citrate synthetase gene product was also amplified using the primer set Rr190.70p-Rr190.602n, which codes for a 532-bp sequence of the 190-kDa surface protein gene (2,10,12,13).Of the 439 ticks tested, 7 were positive by assays with both sets of primers (1.6%). The infection rate by rickettsiae was 2.4% (5 of 207) among R. sanguineus and 1.4% (2 of 138) among R. bursa.…”

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confidence: 99%

First Isolation and Identification of Rickettsia conorii from Ticks Collected in the Region of Fokida in Central Greece

et al. 2003

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Three different spotted-fever group rickettsiae-Rickettsia conorii, R. massiliae, and R. rhipicephali-were detected and identified by PCR-restriction fragment length polymorphism analysis in Rhipicephalus ticks collected from domestic animals in the Fokida region of Greece, where a high seroprevalence of antibodies to R. conorii was previously demonstrated. The infection rate of ticks was 1.6%. Moreover, R. conorii was isolated from one Rhipicephalus sanguineus tick.Rickettsia conorii, the causative agent of Mediterranean spotted fever, is an obligate intracellular, gram-negative bacterium that infects humans. It is transmitted by the bite of infected arthropods (1, 9). The presence of the spotted-fever group (SFG) rickettsiae in ticks has been reported in several countries (2, 6-11, 13). Many rickettsiae remain poorly characterized due to the inability to be maintained in mammalian cell culture, embryonated chicken eggs, or small rodents, which have traditionally been the standard maintenance hosts used in rickettsiology. The advents of new culture techniques, such as the shell vial-centrifugation technique, and the detection of rickettsial DNA have increased the number of rickettsial species identified (6).A seroepidemiological survey, using an immunofluorescence assay and a Western blot method, was conducted in three villages (Leukaditi, Vounichora, and Makrini) of the Greek province of Fokida in 1991. This study reported a high seroprevalence of antibodies to R. conorii in humans (46%) (1). In a culture survey of ticks from the same area, strain GS (for "Greek strain") was isolated. This strain is genetically similar to Rickettsia massiliae; moreover, R. conorii was not found (2). As a follow-up to previous studies, we focused our efforts on determining the presence and infection rate of R. conorii in field-collected ticks from the three study villages of the Fokida (Fig. 1).In the summer of 1998 (May to August), a total of 439 ticks collected from goats, sheep, and dogs were examined for the presence of SFG rickettsiae. According to standard taxonomic keys, 207 ticks were identified as Rhipicephalus sanguineus, 94 were identified as Rhipicephalus turanicus, and 138 were identified as Rhipicephalus bursa (8). Each tick was triturated in 500 l of minimum essential medium supplemented with 4% fetal calf serum and 2 mM L-glutamine. Half of the above suspension was used for the detection of rickettsial DNA by PCR, and the rest was used for the isolation of rickettsiae by the shell vial technique (3, 4, 9, 11-13). The DNA extraction from the ticks was performed using the QIAamp tissue kit (Qiagen, Hilden, Germany), according to the instructions of the manufacturer. The primer set Rp.CS.877-Rp.CS.1258n was used to amplify a 381-bp sequence of the citrate synthase gene (9). As a negative control for each tick sample, pure PCR buffer treated in the same way as the tick samples was included. As a positive control, purified DNA from R. conorii (Moroccan strain) was used. The DNA from ticks that were positive for the c...

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Isolation and Identification of a Rickettsial Strain Related to Rickettsia massiliae in Greek Ticks

Cited by 65 publications

References 0 publications

Simultaneous Detection of “ Rickettsia mongolotimonae ” in a Patient and in a Tick in Greece

Simultaneous Detection of “ Rickettsia mongolotimonae ” in a Patient and in a Tick in Greece

Fourteen-year seroepidemiological study of zoonoses in a Greek village.

First Isolation and Identification of Rickettsia conorii from Ticks Collected in the Region of Fokida in Central Greece

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