Two for the price of one: Co-infection with Rickettsia bellii and spotted fever group Rickettsia in Amblyomma (Acari: Ixodidae) ticks recovered from wild birds in Brazil.

The bacterium Rickettsia bellii has been detected in 25 species of ticks in the American continents, but its pathogenic potential is considered as undetermined. A possible role for this species in the phenomenon of transovarial exclusion of pathogenic members of the spotted fever group (SFG) of Rickettsia has been suggested and co-infections with pathogenic species have been reported infrequently in both North and South America. Traditional methods for the molecular detection of rickettsial agents in ticks focus largely on the identification of sequences found in SFG Rickettsia, an approach that may overlook the presence of co-infections with R. bellii. Two novel, species-specific polymerase chain reaction (PCR) assays, targeting the genes encoding the surface cell antigen (Sca), autotransporter proteins sca9 and sca14, were developed and validated for the detection of R. bellii using 150 Amblyomma ticks collected from wild birds in Brazil. Co-infection of R. bellii infected ticks was evaluated using a novel PCR assay targeting the ompA sequence characteristic of SFG Rickettsia. Preliminary species-level identification was achieved by restriction fragment length polymorphism (RFLP) analysis and subsequently confirmed by sequencing of amplicons. Nine out of seventy-three Amblyomma longirostre and one of two Amblyomma calcaratum ticks were shown to be co-infected with R. bellii and Rickettsia amblyommatis, while two out of sixty-seven Amblyomma sp. haplotype Nazaré ticks were recorded as co-infected with R. bellii and the Rickettsia parkeri-like bacterium, strain ApPR. Interestingly, our data represent the first records of R. bellii in association with A. calcaratum and Amblyomma sp. haplotype Nazaré. The novel PCR-RFLP systems reported herein, provide an alternative, rapid and cost-efficient (relative to strategies based on sequencing or real-time PCR), approach to evaluate rickettsial co-infection of ticks, a potentially significant phenomenon that has most likely been underestimated to date.

[Clinical approach and main tick-borne rickettsiosis present in Latin America]. / Aproximación clínica y principales rickettsiosis transmitidas por garrapatas presentes en Latinoamérica.

Tick-borne rickettsial diseases are potentially life threatening infections that in Latin America have an emerging and reemerging character. Until few years ago, Rickettsia rickettsia was the only tick-borne rickettsia present in America; but nowadays several other species such as R. parkeri and R. massiliae are causing disease in humans in the region. In addition, new species are being described; although their pathogenicity has not been confirmed they should be considered as potential pathogens. Since the microbiological diagnosis of rickettsioses can take days or weeks, a high clinical suspicion and early start of appropriate treatment are crucial. In this review the distribution and main clinical manifestations of tick-borne rickettsial diseases in Latin America are detailed. Since R. felis has been found in ticks and the role of this vector has not been clarified, we have included a section about this pathogen.

Aproximación clínica y principales rickettsiosis transmitidas por garrapatas presentes en Latinoamérica / Clinical approach and main tick-borne rickettsiosis present in Latin America

Tick-borne rickettsial diseases are potentially life threatening infections that in Latin America have an emerging and reemerging character. Until few years ago, Rickettsia rickettsia was the only tick-borne rickettsia present in America; but nowadays several other species such as R. parkeri and R. massiliae are causing disease in humans in the region. In addition, new species are being described; although their pathogenicity has not been confirmed they should be considered as potential pathogens. Since the microbiological diagnosis of rickettsioses can take days or weeks, a high clinical suspicion and early start of appropriate treatment are crucial. In this review the distribution and main clinical manifestations of tick-borne rickettsial diseases in Latin America are detailed. Since R. felis has been found in ticks and the role of this vector has not been clarified, we have included a section about this pathogen.

Las rickettsiosis transmitidas por garrapatas son infecciones potencialmente letales, que en Latinoamérica tienen carácter emergente y re-emergente. Hasta hace escasos años, la única rickettsiosis transmitida por garrapatas era causada por Rickettsia rickettsii, pero en la actualidad existen otras especies como R. parkeri y R. massiliae que están provocando enfermedad en humanos en la región. Por otro lado, se están describiendo candidatos a nuevas especies de Rickettsia, que aunque no han probado su patogenicidad deben considerarse como potencialmente patógenos. Dado que el diagnóstico microbiológico puede tardar días o semanas, resulta fundamental una alta sospecha clínica y la instauración precoz de un tratamiento adecuado. En esta revisión se detalla la distribución y principales manifestaciones clínicas de las rickettsiosis transmitidas por garrapatas en Latinoamérica. Se ha incluido una sección sobre la infección por R. felis, por haberse encontrado esta especie en garrapatas, y no haberse aclarado el papel de este vector en su ciclo epidemiológico.

Syndromic classification of rickettsioses: an approach for clinical practice.

Rickettsioses share common clinical manifestations, such as fever, malaise, exanthema, the presence or absence of an inoculation eschar, and lymphadenopathy. Some of these manifestations can be suggestive of certain species of Rickettsia infection. Nevertheless none of these manifestations are pathognomonic, and direct diagnostic methods to confirm the involved species are always required. A syndrome is a set of signs and symptoms that characterizes a disease with many etiologies or causes. This situation is applicable to rickettsioses, where different species can cause similar clinical presentations. We propose a syndromic classification for these diseases: exanthematic rickettsiosis syndrome with a low probability of inoculation eschar and rickettsiosis syndrome with a probability of inoculation eschar and their variants. In doing so, we take into account the clinical manifestations, the geographic origin, and the possible vector involved, in order to provide a guide for physicians of the most probable etiological agent.

Rickettsiosis transmitidas por garrapatas en las Américas: avances clínicos y epidemiológicos, y retos en el diagnóstico / Tick-borne rickettsioses in the Americas: clinical and epidemiological advances, and diagnostic challenges

Las rickettsiosis son entidades clínicas de tipo zoonótico, causadas por bacterias intracelulares estrictas de los géneros Rickettsia y Orientia, pertenecientes a la familia Rickettsiaceae. Su ecología está determinada por factores ambientales y la presencia de vectores específicos que condicionan el establecimiento y la epidemiología en diferentes regiones del mundo. En las Américas, durante el siglo XX, únicamente eran reconocidas tres de estas enfermedades: la fiebre manchada de las Montañas Rocosas, el tifus epidémico y el tifus endémico, Sin embargo, a partir del año 2000 se han descrito mas de 10 especies diferentes previamente desconocidas en este continente, tanto en artrópodos como en casos clínicos, hecho que permite clasificarlas como entidades clínicas emergentes y reemergentes. Dadas las manifestaciones clínicas de las enfermedades causadas por rickettsias, siendo la gran mayoría inespecíficas y, por lo mismo, compartidas con otras enfermedades infecciosas, especialmente virales y bacterianas, han sido enmarcadas entre los diagnósticos diferenciales del síndrome febril agudo, tanto en áreas urbanas como tropicales. En la actualidad, se cuenta con métodos diagnósticos directos e indirectos, que son útiles en la identificación del agente infeccioso, en este caso, causante de rickettsiosis.

Rickettsioses are a group of zoonotic diseases caused by strict intracellular bacteria of the genus Rickettsia and Orientia which belong to the Rickettsiaceae family. Their ecology is influenced by environmental factors and the presence of specific vectors that determine the establishment and epidemiology in different world regions. In America, during the 20 th century, only three of these diseases were recognized: Rocky Mountain spotted fever, epidemic typhus and endemic typhus. However, since 2000, more than 10 different species that had previously been unknown in this continent have been described, both in arthropods and in clinical cases, fact that classifies them as emerging and re-emerging diseases. Given the clinical manifestations of the diseases caused by rickettsias, being the majority unspecific and, therefore, shared with other infectious diseases, especially viral and bacterial, they have been framed within the differential diagnoses of acute febrile syndrome in urban and tropical areas. Nowadays, there are direct and indirect diagnostic methods, which are useful in the definition of the infectious agent, in this case, the cause of rickettsioses.

[Tick-borne rickettsioses in the Americas: clinical and epidemiological advances, and diagnostic challenges]. / Rickettsiosis transmitidas por garrapatas en las Américas: avances clínicos y epidemiológicos, y retos en el diagnóstico.

Rickettsioses are a group of zoonotic diseases caused by strict intracellular bacteria of the genus Rickettsia and Orientia which belong to the Rickettsiaceae family. Their ecology is influenced by environmental factors and the presence of specific vectors that determine the establishment and epidemiology in different world regions. In America, during the 20 th century, only three of these diseases were recognized: Rocky Mountain spotted fever, epidemic typhus and endemic typhus. However, since 2000, more than 10 different species that had previously been unknown in this continent have been described, both in arthropods and in clinical cases, fact that classifies them as emerging and re-emerging diseases. Given the clinical manifestations of the diseases caused by rickettsias, being the majority unspecific and, therefore, shared with other infectious diseases, especially viral and bacterial, they have been framed within the differential diagnoses of acute febrile syndrome in urban and tropical areas. Nowadays, there are direct and indirect diagnostic methods, which are useful in the definition of the infectious agent, in this case, the cause of rickettsioses.

[An update on rickettsiosis]. / L'actualité des rickettsioses.

Rickettsiae are strictly intracellular bacteria belonging to the Rickettsiaceae family. They are transmitted by various arthropods species, which are either vectors or reservoirs for the bacteria. The specific association between Rickettsieae and the vector are extensively studied, moreover Rickettsieae associated diseases are part of a continuously evolving field. Nevertheless, some rickettsiosis, such as epidemic typhus, have been described since the 16th century. Emerging diseases related to these bacteria are being investigated as well as new species, the implication of which is not always clear in human pathology when they are discovered. Some of these diseases are benign, others may be potentially fatal. Clinicians must therefore be aware of them. These issues are reviewed.

[Far-eastern spotted fever: description of new infectious disease].

Historical data concerning description and identification of etiology of tick-borne spotted fever in Russia are presented in the paper. Review of studies shows that by uncovering the etiology of tick-borne spotted fever in the Russian Far East, an emerging disease may be discovered. Another, different from classical Rickettsia sibirica, bacteria causes spotted fever there. The application of described methods resulted in first isolation of etiological agent of far-eastern spotted fever, Rickettsia heilongjiangensis, strains from patients.

Reemergence of rickettsiosis in Oran, Algeria.

The presumptive cases of Mediterranean spotted fever have been identified in 1993 and since that time, its frequency has steadily increased. The prospective study, in summer 2004, was conducted in order to present the descriptive clinic and epidemiology, to identify more severe forms, the presence of the multiple eschars, and different rickettsial strains caused the disease in our region. In Oran, the cases were diagnosed clinically. In Marseille, serum specimens were tested by IFA using the panel of eight rickettsial antigen; Western blot and cross-adsorption studies were also performed in order to confirm the diagnosis. Ninety-three patients clinically diagnosed were recorded from July 3 to October 28, 2004. Eighty percent were male, the mean age was 44.3 years, 90% were exposed to dog and 32% reported tick bites. Clinical signs were as follow: presence of underlying disease (44%), sudden onset (78%), fever (100%), loss of weight (63%), conjunctivitis (43%), and a tache noire was noticed in 70%. Interestingly, two patients had two and three eschars, respectively. The rash was maculopapular (palm and sole) and purpuric in nine cases. Doxycycline was the most antibiotic (91%) with favourable outcome in 91% of the cases. Malignant form with death is reported for three patients (3.2%). Among the 93 patients, 104 serum from 65 patients were tested (serums of others patients were lost or ticket not found on tube. Sixty-three patients out of 65 had a positive serology by IFA with cross-reactive antibodies especially between R. conorii, R. felis and/or R. typhi. Two others negative serology were: one precocious serum and second from the patient, which presented symptoms of MSF and tested two serums, Western blot and cross-adsorption.

Incidence and distribution pattern of Rickettsia felis in peridomestic fleas from Andalusia, Southeast Spain.

The presence of Rickettsia felis was investigated in three species of pulicid fleas (Ctenocephalides felis, Ctenocephalides canis and Pulex irritans) collected in 38 locales in Andalusia (Spain) over the period 1999-2004. Amplification of a fragment of OmpB gene was positive in 54.17 % of lots of Ct. felis. The identity of the PCR bands was confirmed as R. felis by sequence data obtained directly from the PCR amplicon. No rickettsia was found in Ct. canis nor P. irritans.

Expression of rOmpA and rOmpB protein in Rickettsia massiliae during the Rhipicephalus turanicus life cycle.

Rhipicephalus turanicus tick colony infected in the laboratory with Rickettsia massiliae showed that the rickettsia is transovarially and transdatially tramsmitted. The expression of rOmpB did not change with temperature or the stages of the tick life cycle. In contrast, rOmpA was less expressed during the larval stage.

Proposal to create subspecies of Rickettsia sibirica and an emended description of Rickettsia sibirica.

The Rickettsia sibirica species is composed of isolates that are genotypically close but can be classified within two distinct serotypes, that is, R. sibirica sensu stricto and R. sibirica mongolitimonae (incorrectly named R. mongolotimonae). We investigated the possibility of classifying rickettsiae closely related to R. sibirica as R. sibirica subspecies, as proposed by the ad hoc Committee on Reconciliation of Approaches to Bacterial Systematics. For this, we first estimated the genotypic variability by using multilocus sequence typing (MLST), including the sequencing of five genes, and multispacer typing (MST) using three intergenic spacers, of five isolates and three tick amplicons of R. sibirica sensu stricto and six isolates of R. sibirica mongolotimonae. Then, we selected a representative of each MLST genotype and used mouse serotyping to estimate their degree of taxonomic relatedness. Among the 14 isolates or tick amplicons studied, 2 MLST genotypes were identified: (i) the R. sibirica sensu stricto type; and (ii) the R. sibirica mongolitimonae type. Representatives of the two MLST types were classified within three MST types and into two serotypes. Therefore, as isolates within the R. sibirica species are genotypically homogeneous but show MST genotypic, serotypic, and epidemio-clinical dissimilarities, we propose to modify the nomenclature of the R. sibirica species through the creation of subspecies. We propose the names R. sibirica subsp. sibirica subsp. nov. (type strain = 2-4-6, ATCC VR-541(T)), and R. sibirica subsp. mongolitimonae subsp. nov. (type strain = HA-91, ATCC VR-1526(T)). The description of R. sibirica is emended to accommodate the two subspecies.

Characterisation of rickettsial diseases in a hospital-based population in Malta.

OBJECTIVE: The aim of the study was to characterise the causative agents of rickettsial disease in Malta. A secondary objective was to study the epidemiology of cases of rickettsial disease. METHODS: Cases admitted to St Luke's Hospital between June 2002 and May 2003 presenting with complaints of fever, headache, rash and/or an eschar were considered possible cases of rickettsial disease. A patient interview was conducted within 24h of admission. Paired sera were taken for serology and blood samples sent for rickettsial PCR and culture. Whenever an eschar was present, biopsies were taken for culture and immunohistochemical analysis. RESULTS: Thirty-three cases of possible rickettsial disease were identified. Although serological tests showed cross reactivities between different species of rickettsiae, one was diagnostic for Rickettsia conorii. None of the sera showed any cross-reactivity with Rickettsia typhi. There was one positive biopsy for R. conorii when tested by PCR and another was positive for spotted fever group Rickettsia by immunohistochemistry. CONCLUSION: Spotted fever rickettsiosis is endemic in Malta. Contrary to previous belief, none of the cases were due to murine typhus. The predominant causative agent of rickettsial disease in Malta is likely to be R. conorii, although the animal reservoir has still not been definitely identified.

[Tick borne diseases]. / Durch Zecken übertragene Krankheiten.

It is known for many years that tick-borne diseases have worldwide a high economical impact on farming industry and veterinary medicine. But only in the last twenty years the importance of such diseases were notified in human medicine by the medical community and the public with emerging of the tick borne encephalitis virus and the description of Borrelia burgdorferi. It is often forgotten that many other infectious agents as bacteria, virus, Rickettsia or protozoa can be transmitted by ticks. Such diseases are rarely diagnosed in Europe either they are overlooked and misdiagnosed or they are connected with special professional activities. The development of new regions for tourism with different out door activities (adventure trips, trekking, hunting) leads to an exposure to different tick borne diseases, which are often misdiagnosed.

[Rickettsioses]. / Enfermedades producidas por Rickettsia.

Species of the genus Rickettsia are small, obligate intracelular, gramnegative bacteria, many of which are considered nowadays a paradigm of emergent pathogens. With the exception of R. prowazekii, they are maintained in the natural environment through a cycle involving different hosts (mainly mammals), and arthropod vectors (in general ticks, and fleas); humans are affected only by incidental transmission due to arthropod bites. The common pathogenesis of these diseases lie on the predominantly infection of endotelial cells, that determines the development of multisistemic small vessel vasculitis, which may affect lungs (interstitial pneumonitis), heart (miopericarditis), skin (rash), central nervous system (meningoencephalitis), as well as liver, and kidneys. They are classified in two groups: spotted-fever group, and typhus group rickettsia. In Spain the most prevalent rickettsioses of both groups are mediterranean spotted fever (caused by R. conorii), and murine typhus (caused by R. typhi), respectively. This review focuses mainly in these two diseases, and also in other rickettsioses of interest due to their recently emergence or reemergence (R. slovaca, R. africae, R. prowazekii, R. felis), or to their high incidence in other areas (R. rickettsii, Orientia tsutsugamushi).

[Rickettsioses of the spotted fever-group]. / Rickettsiosen der Zeckenbissfieber-Gruppe.

Rickettsioses are among the most frequent causes of febrile diseases in travelers, as shown in several studies in the last years. Furthermore, since 1991, with the introduction of new testing methods, 8 new Rickettsia species (or new diseases) have been described. Typical symptoms of rickettsial infections are high fever, headaches and myalgias, frequently associated with a rash and/or an inoculation eschar ("tache noire") at the site of tick bite. The rapid recognition of a rickettsiosis and the immediate start of appropriate antibiotic therapy are important because some rickettsioses (e. g. the Rocky Mountain spotted fever) are associated with relevant morbidity and mortality. This paper reviews the spotted fever group rickettsioses.

Rickettsial disease: classical and modern aspects.

Rickettsial diseases have been reassessed in recent years since they represent an important field in today's medicine. New agents have been described: some are non-pathogenic agents and the others are associated with well-defined or peculiar clinical patterns. In addition, different species of rickettsiosis are found in relation to the geographic areas of the world. Some agents may be defined as variants of older diseases whereas most of the newly described forms of rickettsiosis represent distinct entities with unique epidemiologial and clinical features. Probably the main news regards the group of the spotted fevers. An additional new aspect is linked to the medicine of travellers and tourists. However, this aspect may not be significant for the rickettsial diseases in relation to other human illnesses, such as malaria. Therefore, an investigation into the geographical origin of patients has to enter our routine medical work.

[Tick-borne rickettsioses of domestic ruminants: taxonomic elements and laboratory diagnosis]. / Les rickettsioses des ruminants domestiques transmises par les tiques: elements taxonomiques et diagnostic de laboratoire.

Tick-borne rickettsioses are diseases which remain comparatively unknown to field veterinarians. The authors present a literature review, focusing on taxonomic aspects and on laboratory diagnosis.