Bartonella quintana, lice, and molecular tools.
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Publications and source records attributed to Pierre-Edouard Fournier.
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African tick-bite fever (ATBF) is a rickettsiosis caused by Rickettsia africae. We describe histologic features and immunodetection of R. africae in cutaneous inoculation eschars from 8 patients with ATBF, which was diagnosed by culture or association of positive PCR detection and positive serologic results. We used quantitative image analysis to compare the pattern of inflammation of these eschars with those from Mediterranean spotted fever. We evaluated the diagnostic value of immunohistochemical techniques by using a monoclonal antibody to R. africae. ATBF eschars were histologically characterized by inflammation of vessels composed mainly of significantly more polymorphonuclear leukocytes than are found in cases of Mediterranean spotted fever (p < 0.05). Small amounts R. africae antigens were demonstrated by immunohistochemical examination in 6 of 8 patients with ATBF. Neutrophils in ATBF are a notable component of the host reaction, perhaps because ATBF is a milder disease than the other rickettsioses. Immunohistochemical detection of rickettsial antigens may be useful in diagnosing ATBF.
The prevalences of Bartonella spp. and Rickettsia spp. were investigated using molecular methods in 77 rodent fleas collected in November 2002 by the French forces detachment in Kabul, Afghanistan. Overall, Bartonella DNA was detected in 15.5% of gerbil fleas and 40.5% of rat fleas, whereas Rickettsia felis was found in 9% of gerbil fleas. We described for the first time in this country Bartonella quintana, B. koehlerae, B. taylorii, and Rickettsia felis in fleas from the gerbil species Meriones lybicus, and B. elizabethae and B. doshiae in rat fleas. Of these, B. quintana, B. elizabethae, B. koehlerae, and R. felis are recognized human pathogens. These results emphasize the potential risk of flea-borne infections transmitted by rodents in this area, and suggest that preventive measures should be taken in the general framework of zoonoses management.
Thirty-one rickettsial isolates from ticks or patients in North Asian tick typhus (NATT) foci from the Ural region to the Russian Far East were obtained at the Omsk Research Institute of Natural Foci Infections between 1954 and 2001. Using citrate synthase (gltA) and outermenbrane protein a (ompA) gene sequencing, we identified these isolates as Rickettsia sibirica sensu stricto (25 isolates), R. sibirica strain BJ-90 (2 isolates), R. slovaca (1 isolate), and R. heilongjiangensis (3 isolates). We demonstrate that Ixodes persulcatus ticks should be considered potential vectors of NATT. We also demonstrate the presence of R. slovaca in Ural and R. heilongjiangensis in Siberia and Russian Far East, where they may cause human infections misdiagnosed as cases of NATT. Clinicians should be aware that several spotted fever rickettsioses with different prognoses coexist in Russia in areas where NATT was the only previously recognized rickettsiosis.
BACKGROUND: Many soldiers in Napoleon's Grand Army died of infectious diseases during its retreat from Russia. Because soldiers were commonly infested with body lice, it has been speculated that louse-borne infectious diseases, such as epidemic typhus (caused by Rickettsia prowazekii), were common. METHODS: We investigated this possibility during recent excavations of a mass grave of Napoleon's soldiers in Vilnius, Lithuania. Segments of 5 body lice, identified morphologically and by polymerase chain reaction (PCR) amplification and sequencing, were found in earth from the grave that also contained fragments of soldiers' uniforms. RESULTS: DNA of Bartonella quintana (the agent of trench fever) was identified by PCR and sequencing in 3 of the lice. Similarly, PCR and sequencing of dental pulp from the remains of 35 soldiers revealed DNA of B. quintana in 7 soldiers and DNA of R. prowazekii in 3 other soldiers. CONCLUSIONS: Our results show that louse-borne infectious diseases affected nearly one-third of Napoleon's soldiers buried in Vilnius and indicate that these diseases might have been a major factor in the French retreat from Russia.
BACKGROUND: Pseudomonas luteola has been recognized as an uncommon cause of bacteremia and of infections in patients with underlying medical disorders. CASE PRESENTATION: We isolated P. luteola from blood cultures in a patient with prosthetic valve endocarditis developed 16 months after cardiac surgery. CONCLUSION: P. luteola is a rare opportunistic agent, with a propensity of infecting valvular prostheses.
We sequenced the genome of Rickettsia felis, a flea-associated obligate intracellular alpha-proteobacterium causing spotted fever in humans. Besides a circular chromosome of 1,485,148 bp, R. felis exhibits the first putative conjugative plasmid identified among obligate intracellular bacteria. This plasmid is found in a short (39,263 bp) and a long (62,829 bp) form. R. felis contrasts with previously sequenced Rickettsia in terms of many other features, including a number of transposases, several chromosomal toxin-antitoxin genes, many more spoT genes, and a very large number of ankyrin- and tetratricopeptide-motif-containing genes. Host-invasion-related genes for patatin and RickA were found. Several phenotypes predicted from genome analysis were experimentally tested: conjugative pili and mating were observed, as well as beta-lactamase activity, actin-polymerization-driven mobility, and hemolytic properties. Our study demonstrates that complete genome sequencing is the fastest approach to reveal phenotypic characters of recently cultured obligate intracellular bacteria.
The Rickettsia genus is a group of obligate intracellular parasitic alpha-proteobacteria that includes human pathogens responsible for the typhus disease and various types of spotted fevers. rOmpA and rOmpB are two members of the "surface cell antigen" (Sca) autotransporter (AT) protein family that may play key roles in the adhesion of the Rickettsia cells to the host tissue. These molecules are likely determinants for the pathogenicity of the Rickettsia and represent good candidates for vaccine development. We identified the 17 members of this family of outer-membrane proteins in nine fully sequenced Rickettsia genomes. The typical architecture of the Sca proteins is composed of an N-terminal signal peptide and a C-terminal AT domain that promote the export of the central passenger domain to the outside of the bacteria. A characteristic of this family is the frequent degradation of the genes, which results in different subsets of the sca genes being expressed among Rickettsia species. Here, we present a detailed analysis of their phylogenetic relationships and evolution. We provide strong evidence that rOmpA and rOmpB as well as three other members of the Sca protein family--Sca1, Sca2, and Sca4--have evolved under positive selection. The exclusive distribution of the predicted positively selected sites within the passenger domains of these proteins argues that these regions are involved in the interaction with the host and may be locked in "arms race" coevolutionary conflicts.
BACKGROUND: Rickettsia sibirica mongolotimonae has been found in Hyalomma ticks in Inner Mongolia (in China) and Niger and in humans in France and South Africa. To date, only 3 cases of human infection have been reported. METHODS: Patients received a diagnosis of R. sibirica mongolotimonae infection on the basis of culture and/or PCR results plus serological test results. RESULTS: From January 2000 to June 2004, R. sibirica mongolotimonae infection was diagnosed in 7 patients. In 3 patients, the bacterium was cultivated from the inoculation eschar. The other 4 patients had cases that were diagnosed with use of PCR of samples obtained from the eschar (2 patients) or blood (2 patients), plus specific Western blot before (2 patients) and after (2 patients) cross-adsorption. The clinical presentation included fever (temperature, >38.5 degrees C), a maculopapular rash, and > or =1 inoculation eschar in 6 patients, enlarged regional lymph nodes in 4 patients, and lymphangitis in 3 patients. On the basis of the study of 9 cases, R. sibirica mongolotimonae infection differed from other tick-borne rickettsioses in the Mediterranean area in the following ways: it involved a specific incidence in the spring, the presence of 2 eschars in 2 (22%) of the patients, the presence of a draining lymph node in 5 (55%) of the patients, and lymphangitis expanding from the inoculation eschar to the draining node in 4 (44%) of the patients. The most recent patient in our series received a clinical diagnosis on the basis of such findings. All patients recovered without any sequelae. CONCLUSIONS: We propose that this new rickettsiosis be named "lymphangitis-associated rickettsiosis." Lymphangitis-associated rickettsiosis should be considered in the differential diagnosis of tick-borne rickettsioses in Europe, Africa, and Asia.
BACKGROUND: Rickettsiae closely related to the Malish strain, the reference Rickettsia conorii strain, include Indian tick typhus rickettsia (ITTR), Israeli spotted fever rickettsia (ISFR), and Astrakhan fever rickettsia (AFR). Although closely related genotypically, they are distinct serotypically. Using multilocus sequence typing (MLST), we have recently found that distinct serotypes may not always represent distinct species within the Rickettsia genus. We investigated the possibility of classifying rickettsiae closely related to R. conorii as R. conorii subspecies as proposed by the ad hoc committee on reconciliation of approaches to bacterial systematics. For this, we first estimated their genotypic variability by using MLST including the sequencing of 5 genes, of 31 rickettsial isolates closely related to R. conorii strain Malish, 1 ITTR isolate, 2 isolates and 3 tick amplicons of AFR, and 2 ISFR isolates. Then, we selected a representative of each MLST genotype and used multi-spacer typing (MST) and mouse serotyping to estimate their degree of taxonomic relatedness. RESULTS: Among the 39 isolates or tick amplicons studied, four MLST genotypes were identified: i) the Malish type; ii) the ITTR type; iii) the AFR type; and iv) the ISFR type. Among these four MLST genotypes, the pairwise similarity in nucleotide sequence varied from 99.8 to 100%, 99.4 to 100%, 98.2 to 99.8%, 98.4 to 99.8%, and 99.2 to 99.9% for 16S rDNA, gltA, ompA, ompB, and sca4 genes, respectively. Representatives of the 4 MLST types were also classified within four types using MST genotyping as well as mouse serotyping. CONCLUSION: Although homogeneous genotypically, strains within the R. conorii species show MST genotypic, serotypic, and epidemio-clinical dissimilarities. We, therefore, propose to modify the nomenclature of the R. conorii species through the creation of subspecies. We propose the names R. conorii subsp. conorii subsp. nov. (type strain = Malish, ATCC VR-613), R. conorii subspecies indica subsp. nov. (type strain = ATCC VR-597), R. conorii subspecies caspia subsp. nov. (type strain = A-167), and R. conorii subspecies israelensis subsp. nov. (type strain = ISTT CDC1). The description of R. conorii is emended to accomodate the four subspecies.
BACKGROUND: Histological demonstration of microorganisms, vegetations, or active endocarditis in cardiac valve tissue is included in the Duke criteria and is considered to be a criterion of confirmed infective endocarditis. However, the histological features that characterize infective endocarditis are not accurately defined at the qualitative and quantitative levels. METHODS: Pathologic analysis of tissue adjoining mechanical cardiac valves was undertaken retrospectively for 21 patients who underwent surgical removal of a mechanical valve because of suspected infective endocarditis and 69 patients who underwent surgical removal of a mechanical valve because of noninfectious dysfunction. To better define the histological criteria for infective endocarditis, we used quantitative image analysis to compare these 2 groups of patients with respect to valvular fibrosis, calcifications, vegetations, patterns of inflammation, and vascularization. RESULTS: Histologically, infective endocarditis in patients with mechanical valves was characterized by the demonstration of microorganisms, vegetations, and significant neutrophil-rich inflammatory infiltrates with extensive neovascularization. In contrast, valve tissue specimens from patients with mechanical valves that were removed because of noninfectious complications showed significant rates of extensive fibrosis and, when present, inflammatory infiltrates that were mainly composed of macrophages and lymphocytes. A neutrophil surface area with a cutoff value of > or =2% of the total valve tissue surface is highly predictive of (90%) and specific for (98%) infective endocarditis. CONCLUSIONS: When no microorganisms are detected and vegetations are lacking in tissue adjacent to a mechanical valve, neutrophil-rich inflammation and extensive neovascularization might better histologically define the term "active endocarditis" in the Duke criteria. This definition would allow differentiation between infective endocarditis and inflammatory noninfectious valve processes in patients with mechanical cardiac valves.
Recent studies have shown that Rickettsia typhi, R. felis, Bartonella henselae, and B. clarridgeiae occur in New Zealand. To raise awareness of these emerging and re-emerging flea-borne bacterial pathogens among New Zealand health workers, we review the clinical features, diagnosis, and treatment of infections, and the biology of the major flea vectors.
PURPOSE: Histologic examination of valve samples is considered as the gold standard for the diagnosis of infectious endocarditis. Molecular tools are also very promising for patients with negative-culture endocarditis. Thus, we studied the contribution of valvular histology, culture, and 16S rRNA PCR amplification plus sequencing to the diagnosis of infectious endocarditis in patients undergoing valve surgery. SUBJECTS AND METHODS: We performed culture, histological examination, and broad-range PCR amplification plus sequencing on valve samples taken from 127 patients with infectious endocarditis and from 118 patients without endocarditis. The sensitivity and specificity of these tests for the diagnosis of endocarditis in patients undergoing valve surgery were studied. RESULTS: The sensitivity of PCR was of 61% (64/105) whereas that of histological examination was of 63% (62/98) and that of valve culture was of only 13% (14/105). All 68 positive PCR results considered reliable according to an interpretation scheme were from patients with infectious endocarditis, resulting in a 100% (118/118) specificity of the interpreted molecular approach. The specificity of histology was also of 100% (118/118) when using stringent criteria (ie, presence of vegetation, microorganisms, and/or valvular inflammation with mainly polymorphonuclear cells). PCR identified an etiological agent in 38% (5/13) of definite blood culture-negative infectious endocarditis. CONCLUSION: We show that valvular histology with stringent criteria is the gold standard for the diagnosis of infectious endocarditis. Broad-range amplification of 16S rRNA gene is indicated for infectious endocarditis of unknown etiology, whereas valve culture is of limited sensitivity.
Thirty strains of Borrelia burgdorferi sensu lato have been isolated from Ixodes persulcatus ticks and from skin lesions of Lyme disease patients in the Russian Far East from 1997 to 2003. We amplified full-length outer surface protein A (ospA) gene of all strains. BLAST search and following phylogenetic analysis showed that strains form four well-defined groups. Four strains belong to Borrelia afzelii species. Other strains distributed into tree major groups, identified as Borrelia garinii. Indeed, based on the ospA gene comparison, phylogenetic relationship of these groups among each other does not differ from relationship among other previously defined groups inside B. burgdorferi sensu lato genogroup, such as B. afzelii or Borrelia bissettii. Further investigations of genetic and serologic properties of the strains belonging to those groups are required in order to clarify their taxonomic status.
A chromosomally encoded oxacillinase, OXA-69, was characterized from Acinetobacter baumannii AYE. beta-Lactamase OXA-69 shared 97% amino acid identity with the recently described OXA-51 enzyme of A. baumannii and 62 and 56% amino acid identity with the carbapenem-hydrolyzing oxacillinases OXA-24 and OXA-23, respectively. Biochemical characterization of the purified OXA-69 revealed a narrow-spectrum hydrolysis profile but including, at a low level, imipenem and meropenem. By PCR and sequencing bla(OXA-69)-like genes were identified in all A. baumannii strains tested (n = 12), suggesting that this oxacillinase is naturally occurring in that species.
Canine DNA samples from South Africa were found to contain 16S rRNA gene nucleotide and citrate synthase gene nucleotide and deduced amino acid sequences that were most similar to Anaplasma phagocytophilum: 98%, 66%, and 69% similarity, respectively. This suggests that a new Anaplasma species closely related to A. phagocytophilum occurs in Africa.
Currently, there is no tool for typing Rickettsia prowazekii, the causative agent of epidemic typhus, currently considered a potential bioterrorism agent, at the strain level. To test if the multispacer typing (MST) method could differentiate strains of R. prowazekii, we amplified and sequenced the 25 most variable intergenic spacers between the R. prowazekii and R. conorii genomes in five strains and 10 body louse amplicons of R. prowazekii from various geographic origins. Two intergenic spacers, i.e., rpmE/tRNA(fMet) and serS/virB4, were variable among tested R. prowazekii isolates and allowed identification of three and two genotypes, respectively. When the genotypes obtained from the two spacers were combined, we identified four different genotypes. MST demonstrated that several R. prowazekii strains circulated in human body lice during an outbreak of epidemic typhus in Burundi. This may help to discriminate between natural and intentional outbreaks. Our study supports the usefulness of MST as a versatile method for rickettsial strain genotyping.