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The Bartonella henselae sucB gene encodes a dihydrolipoamide succinyltransferase protein reactive with sera from patients with cat-scratch disease.

Bartonella henselae is a recently recognized pathogenic bacterium associated with cat-scratch disease, bacillary angiomatosis and bacillary peliosis. A recombinant clone expressing an immunoreactive antigen of B. henselae was isolated by screening a genomic DNA cosmid library by Western blotting with sera pooled from patients positive for B. henselae IgG antibodies by indirect immunofluorescence (IFA). The deduced amino acid sequence of the 43.7 kDa encoded protein was found to be 76.3 % identical to the dihydrolipoamide succinyltransferase enzyme (SucB) of Brucella melitensis. SucB has been shown to be an immunogenic protein during infections by Brucella melitensis, Coxiella burnetii and Bartonella vinsonii. The agreement between reactivity with a recombinant SucB fusion protein on immunoblot analysis and the results obtained by IFA was 55 % for IFA-positive sera and 88 % for IFA-negative sera. Cross-reactivity was observed with sera from patients with antibodies against Brucella melitensis, Mycoplasma pneumoniae, Francisella tularensis, Coxiella burnetii and Rickettsia typhi.

Acyltransferases↗

Analysis of caprine IgG1 and IgG2 subclass responses to Chlamydia psittaci infection and vaccination.

Enzyme-linked immunosorbent assays (ELISAs) specific for caprine IgG(H+L), IgG1 and IgG2 were developed and evaluated for serodiagnosis of Chlamydia psittaci infections in a Tunisian goat flock with currently occurring chlamydial abortions and a clinically inapparent goat flock of an animal research facility. Additionally, ELISAs were applied to record the IgG1 and IgG2 dynamics of four goats vaccinated with inactivated Chlamydia psittaci and Coxiella burnetii. For screening purposes, the IgG(H+L) ELISA proved to be superior to the complement fixation test because it detected a larger number of chlamydial abortions and was easier to perform and to interpret. Analysis of Chlamydia psittaci-specific IgG1 and IgG2 responses to naturally occurring infections by ELISA revealed high IgG1 levels associated with IgG2 in goats with current abortions, whereas clinically inapparent, but seropositive goats were characterized by significantly lower IgG1 levels only (P less than 0.001). Similarly, the four vaccinated goats responded initially with Chlamydia psittaci-specific IgG1, whereas second and third vaccinations induced (as in goats with chlamydial abortions) predominantly IgG1, but also IgG2. The results indicated that clinically inapparent chlamydial infection may be distinguished from overt disease by analysing specific IgG1 and IgG2 responses. Applying Coxiella burnetii- specific ELISAs on field samples, IgG1 alone could be detected in eight, IgG2 alone in one and IgG1 combined with IgG2 in nine goats. The coxiella-specific antibody response of the four vaccinated goats was--in contrast to the chlamydia-specific response--characterized by IgG2 dominance.

Abortion, Veterinary↗

[Meningo encephalitis revealing Q fever: two cases and a review of the literature].

Q fever is a zoonosis caused by Coxiella burnetii. The acute Q fever is usually characterized by a self-limited flu-like syndrome, fever, pneumonia and hepatitis. Symptoms of the chronic Q fever (evolution>3 months) mainly consist of endocarditis with negative culture. Focal neurological symptoms are rarely observed. Neurological symptoms of acute Q fever consist of meningitis or meningo-encephalitis. Neurological symptoms of chronic Q fever are cerebral embolisms from Coxiella burnetii infected heart valves. We herein report two patients with meningoencephalitis revealing acute Q fever.

Adolescent↗

Surveillance of Egyptian fleas for agents of public health significance: Anaplasma, Bartonella, Coxiella, Ehrlichia, Rickettsia, and Yersinia pestis.

Serologic surveys in Egypt have documented human and animal exposure to vector-borne bacterial pathogens, but the presence and distribution of these agents in arthropods has not been determined. Between July 2002 and July 2003, fleas were collected from 221 mammals trapped in 17 cities throughout Egypt. A total of 987 fleas were collected, representing four species (Ctenocephalides felis, Echidnophaga gallinacea, Leptopsylla segnis, and Xenopsylla cheopis); 899 of these fleas were X. cheopis from rats (Rattus spp.). Fleas were tested for DNA from Anaplasma spp., Bartonella spp., Coxiella burnetii, Ehrlichia spp., Rickettsia spp., and Yersinia pestis. Rickettsia typhi, the agent of murine typhus, was detected in X. cheopis and L. segnis from rats from nine cities. A spotted-fever group Rickettsia sp. similar to "RF2125" was detected in E. gallinacea, and two unidentified spotted fever group Rickettsia were detected in two X. cheopis. Novel Bartonella genotypes were detected in X. cheopis and L. segnis from three cities. Coxiella burnetii was detected in two fleas. Anaplasma, Ehrlichia, and Y. pestis were not detected.

Animals↗

Rickettsial agents in Egyptian ticks collected from domestic animals.

To assess the presence of rickettsial pathogens in ticks from Egypt, we collected ticks from domestic and peridomestic animals between June 2002 and July 2003. DNA extracts from 1019 ticks were tested, using PCR and sequencing, for Anaplasma spp., Bartonella spp., Coxiella burnetii, Ehrlichia spp., and Rickettsia spp. Ticks included: 29 Argas persicus, 10 Hyalomma anatolicum anatolicum, 55 Hyalomma anatolicum excavatum, 174 Hyalomma dromedarii, 2 Hyalomma impeltatum, 3 Hyalomma marginatum rufipes, 55 unidentified nymphal Hyalomma, 625 Rhipicephalus (Boophilus) annulatus, 49 Rhipicephalus sanguineus, and 17 Rhipicephalus turanicus. Ticks were collected predominantly (>80%) from buffalo, cattle, and camels, with smaller numbers from chicken and rabbit sheds, sheep, foxes, a domestic dog, a hedgehog, and a black rat. We detected Anaplasma marginale, Coxiella burnetii, Rickettsia aeschlimannii, and four novel genotypes similar to: "Anaplasma platys," Ehrlichia canis, Ehrlichia spp. reported from Asian ticks, and a Rickettsiales endosymbiont of Ixodes ricinus.

Animals↗

Clinical, epidemiological and epizootic features of Q fever in the northern coastal part of Croatia from 1989 to 1998.

Although Q fever is endemic in the northern coastal part of Croatia, it usually occurs sporadically. Analysis of 58 patients hospitalized for Q fever during the last 10-year period (1989-1998) revealed some differences in the clinical manifestation compared to a previous study (1954-1977). Most cases of Q fever (N = 55; 91%), presented with pneumonia, but no rash was noticed, compared to 46% of patients with Q fever developing exanthema in the previous study. The previously observed high seropositivity to Coxiella burnetii among domestic animals was confirmed in this study. A two-peak seasonal distribution of Q fever observed in 1991 was connected with the imported Russian sort of sheep with special biology of delivery. The clinical outcome was favorable for all patients, since no complications or chronic forms of the disease were recorded. Disproportion between the number of registered and hospitalized patients, including a number of asymptomatic and several undiagnosed or misdiagnosed infections, leads to a conclusion that the real number of persons infected with Coxiella burnetii in the area is several times higher.

Adolescent↗

[Epidemiological investigations transmisssion of Q fever among humans in Bosnia and Herzegovina].

Appearance of an acute disease by Coxiella burnetii in humans in Bosnia and Herzegovina is very big epidemiological problem because incidence rate is among the highest incidence rates in Europe. Incidence rate of Q fever in Federation Bosnia and Herzegovina was very high during 1998. year 5.68% and the highest in 2000, with 115 diseases with acute clinical form of the disease and incidence rate of 6.95%. The incubation period varied from 9 to 28 days. Real incidence rate because of asymptomatic appearance of the disease is very much higher, at least 40-50 times higher. Chronic infection with Coxiella burnetii has been the most common developed as infectious endocarditis, which is the hardest complication. The researches of mode of transmission Q fever in humans showed that this disease spreads in epidemic and sporadic form in Bosnia and Herzegovina. In the period 1998-2003 year, there were 373 cases of Q fever, among them 265 or 71,05% cases epidemic transmission, and 108 or 28,95% cases sporadic. The research of outbreaks of Q fever showed that appearance of Q fever is statistical significant with risk factors at 266 cases of disease. The mode of transmission in 107 cases was the most probably across inhalation of infected aerosols.

Acute Disease↗

Molecular diagnosis of infective endocarditis by PCR amplification and direct sequencing of DNA from valve tissue.

We used broad-range eubacterial PCR amplification followed by direct sequencing to identify microbial pathogens in heart valve material from 29 patients with histologically confirmed infective endocarditis and 23 patients free of infective endocarditis. Microorganisms cultured by conventional techniques matched those identified by PCR in 21 cases. PCR alone identified the causative agent in three cases (Streptococcus bovis, Staphylococcus cohnii, and Coxiella burnetii), allowing better patient management. PCR corrected the initial bacteriological diagnosis in three cases (Streptococcus bovis, Streptococcus mutans, and Bartonella henselae). Among the 29 cases of histologically confirmed infective endocarditis, PCR findings were positive in 27 cases and were consistent with the bacterial morphology seen at Gram staining (26 cases) or with the results obtained by immunohistologic analysis with an anti-C. burnetii monoclonal antibody (one case). In two other cases of histologically confirmed infective endocarditis, PCR remained negative in a blood culture-negative case for which no bacteria were seen at histological analysis and in one case with visualization of cocci and blood cultures positive for Enterococcus faecalis. Ten clinical diagnoses of possible infective endocarditis were ruled out by histopathological analysis of the valves and subsequently by PCR. PCR was negative in 13 of the 14 patients in whom infective endocarditis was rejected on clinical grounds; the other patient was found to have Coxiella burnetii infective endocarditis on the basis of PCR and histopathological analysis and was subsequently included in the group of 29 definite cases. In total, PCR contributed to the diagnosis and management of infective endocarditis in 6 of 29 (20%) cases.

DNA, Bacterial↗

Serological investigations for some bacterial and viral pathogens in fallow deer (Cervus dama) and wild boar (Sus scrofa) of the San Rossore Preserve, Tuscany, Italy.

Sera of 43 fallow deer (Cervus dama) of the San Rossore Preserve (Tuscany, Italy) were examined for antibodies against eight pathogens; one proved positive for Brucella sp., 21 for Listeria monocytogenes, 34 for Chlamydia psittaci, three for Coxiella burnetii, one for infectious bovine rhinotracheitis virus, 11 for parainfluenza-3 virus, 25 for bovine viral diarrhea virus and six for bovine respiratory syncytial virus. No age and sex difference in the positivity rates and titers was evidenced, while a sex difference was found both in rates of infection and in titers against parainfluenza-3 virus. Parainfluenza-3 infection was more prevalent in 1984 than in 1983 sampling. Sera of 20 wild boars (Sus scrofa) of the same preserve were examined for antibodies against five pathogens: four sera were positive for Brucella sp., while all were negative for Listeria monocytogenes, Chlamydia psittaci, Coxiella burnetii and Aujeszky's disease virus. Public and animal health involvement with these diseases are discussed for these respective host species.

Animals↗

[Seroprevalence of human Q fever in Switzerland].

In current practice, only serology, by the titration of anti-Coxiella burnetii antibodies, permits the diagnosis of acute or chronic Q fever. New serological techniques have also made it possible to more accurately determine the number of people in a community who have been affected by this infectious agent. We have thus been able to compare the results of 1437 specimens from the whole of Switzerland with 4009 taken in the Valais. The analysis of these sera by indirect microimmunofluorescence now shows that Q fever is much more prevalent than was generally estimated. However, these results vary greatly from one area and another, between 6.7% and 31.7% depending on the locality. Normally the lowest percentages were found in the big cities (from 9.5% to 12.2% in Berne, Geneva and Zurich) whilst the highest incidences were found in mountain regions (from 23.6% to 31.7% in the Tessin, Uri or the Valais). Regarding sex distribution, it was found that the incidence of Q fever is on average slightly lower (between 3% and 5%) in women than in men. It was also noted that the percentage of people having anti-Coxiella burnetii antibodies increases with age. The high incidence of Q fever in Switzerland is also of concern because of a severe and chronic form of this infection: endocarditis.

Adolescent↗

Microbial aetiology of community-acquired pneumonia in hospitalised patients.

Adult patients hospitalised with community-acquired pneumonia were studied prospectively to determine the microbial aetiology of pneumonia. Between April 1996 and March 1997, blood and sputum samples were collected for culture. Throat swabs were obtained for isolation of viruses and for detection of antigens of Chlamydia pneumoniae, influenza viruses A and B, respiratory syncytial virus and parainfluenza virus. Antibodies against Legionella spp., Mycoplasma pneumoniae, Chlamydia pneumoniae, Chlamydia psittaci, Coxiella burnetii, influenza viruses A and B, respiratory syncytial virus, adenovirus and parainfluenza virus were tested in serum samples. Two hundred eleven patients were included in the study; paired sera were available from 152 patients. Blood culture was positive in 23 (10.9%) patients, Streptococcus pneumoniae being the bacterium isolated most frequently. A fourfold or greater rise or fall in the Chlamydia pneumoniae IgG and/or IgM antibody titre was found in 20 (9.5%) patients and a high antibody titre (> or = 1:512) in the first and/or the second serum sample in 18 (18.5%) patients. Antibodies confirming acute Mycoplasma pneumoniae infection were found in 12 (5.7%) patients, Legionella spp. in six (2.8%), Chlamydia psittaci in two and Coxiella burnetii in one. Three patients had pulmonary tuberculosis. Only two patients had a virus present in the throat swab (adenovirus in one patient and echovirus in the other), and in nine patients, viral antigen was detected. Acute viral infection was confirmed in 51 (24.1%) patients. Bacterial pneumonia was diagnosed in 84 (39.8%) patients, 23 of whom had concurrent viral infection. Acute viral pneumonia without any other identified pathogen was diagnosed in 28 patients. Streptococcus pneumoniae and Chlamydia pneumoniae were the most frequently identified microorganisms.

Aged↗

Hyperendemic focus of Q fever related to sheep and wind.

Q fever is a worldwide zoonosis which is caused by Coxiella burnetii and presents as both acute or chronic cases. The disease can be transmitted from animal reservoirs to humans by the inhalation of infected aerosols. The authors investigated the epidemiology of Q fever in the Bouches-du-Rhône district of southern France. The study area was centered around the small town of Martigues near the cities of Marseille and Aix-en-Provence, where the incidence of the disease seemed higher than in neighboring areas. Epidemiologic data included sheep breeding and wind. Between 1990 and 1995, Q fever was diagnosed in 289 patients, leading to an incidence rate of 35.4 per 100,000 in the study area (range: 6-132), compared with 6.6 in the area of Marseille, and 11.4 in the area of Aix-en-Provence. There was a graphical and statistical relation between the sheep densities, the incidence of the disease, and the strong, local wind known as the Mistral, which blows from the northwest. Although Coxiella burnetii transmission is multifactorial, we may speculate that the high endemicity in the study area is related to a contamination by aerosols because the Mistral blows through the local steppe where 70,000 sheep are bred. This public health problem requires further studies in order to confirm this hypothesis, and to identify more individual and preventable risk factors.

Adolescent↗

Correlation of DNA, RNA, and protein content by flow cytometry in normal and Coxiella burnetii-infected L929 cells.

L929 cell populations persistently infected for more than 600 days exhibited cell cycle distributions and generation times similar to those of uninfected cells. The RNA and protein contents of these long-term-infected cells, as determined by flow cytometry and correlated with the cell cycle, were likewise approximately the same as those in normal uninfected cells. Also, the ratios of RNA to DNA and RNA to protein of infected cells were not significantly different from those of normal cells. Collectively, these data indicate that the long-term-infected cells remained in a state of balanced growth and maintained normal cycle progression and division capacity, allowing for growth and proliferation of host cells and thus ensuring the propagation and persistence of the parasite.

Animals↗

Improved procedure for the drying and storage of polyacrylamide slab gels.

An improved procedure for the drying and storage of polyacrylamide (PAA) slab gels of various sizes and acrylamide contents is presented. A PAA slab gel is preferentially fixed in methanol-acetic acid-glycerol-water (1.0:1.0:0.1:2.0) and then dried under low pressure in an adapted photoprint dryer between Cellophane and polyethylene foils at 75-80 degrees C for 50-90 min. Strong, transparent and flexible sheets are obtained that can be stored for extended periods. The potential of the procedure is exemplified following electrophoresis of both lipopolysaccharides and protein isolated from Coxiella burnetii bacterium.

Bacterial Proteins↗

Serological surveillance of brucellosis and Q fever in cattle in the Central African Republic.

No data are available concerning the seroprevalence of brucellosis in Central African Republic (CAR) and the last report concerning the seroprevalence of Q fever in CAR is from 1995. The aim of our study was to determine the prevalence of these diseases in CAR, especially in Zebu cattle. We used the Rose Bengal Plate Test to test 2032 bovine serum samples for antibodies to Brucella spp. and an indirect immunofluorescence assay to test 784 bovine serum samples for antibodies to Coxiella burnetii (the species responsible for Q fever). The mean seroprevalences of antibodies to Brucella and Coxiella were 3.3 and 14.3%, respectively. Significant differences were found between regions and herds for both diseases. However, relation with differences of climate or vegetation were not evident. Therefore, further data are necessary to better understand the epidemiology of these diseases in CAR and evaluate losses to the farmers.

Agglutination Tests↗

Activity of telithromycin against thirteen new isolates of C. burnetii including three resistant to doxycycline.

In this study we have evaluated the in vitro activity of antibiotics against 13 new isolates of Coxiella burnetii using a real-time quantitative PCR assay. MICs against doxycycline ranged from 1 to 8 microg/mL, telithromycin from 0.5 to 2 microg/mL, and all strains had MICs > or = 8 microg/mL for erythromycin. We report that strains resistant to doxycycline exist either in humans or animals.

Animals↗

DNA probes for the identification of Coxiella burnetti strains.

Isolation of Coxiella Burnetii in the standard laboratory setting is hazardous; therefore most diagnoses are based on retrospective detection of a rising antibody titer to C. burnetti. As a result, this disease is usually undiagnosed or misdiagnosed. Methods for the rapid detection of C. burnetti have now been developed that utilize specific hybridization of labeled DNA probes to nucleic acid in clinical samples. One method detects the presence of C. burnetii 16S ribosomal RNA (rRNA); another uses plasmid sequences. We have developed a probe that detects C. burnetii and one that differentiates between Coxiella strains capable of causing chronic disease and those that cause the acute form. Using these probes, C. burnetii can be identified in blood, urine, and tissue samples. The plasmid-derived probes detect as few as 10(4) organisms and less than 1 ng of Coxiella DNA. A third method differentiates between chronic (endocarditis-causing) strains and those that cause acute Q fever. This method uses the polymerase chain reaction (PCR), in which the target regions of DNA are amplified by iterative cycles of Taq I DNA polymerase chain extension to produce up to a 10(6) amplification of the target sequences. When Southern blotting is used in conjunction with PCR, the test detects as few as 2-9 C. burnetti cells.

Animals↗