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Experimental infection of hare (Lepus europaeus) with Coxiella burnetii and Rickettsia slovaca.

Hares (Lepus europaeus L.) infected subcutaneously (s.c.) or per os with Coxiella burnetii and s.c. with Rickettsia slovaca overcame an inapparent infection accompanied by irregular rickettsaemia and distribution of rickettsiae to different organs and variable antibody response. Neither riskettsia could be detected in faeces of infected animals, but C. burnetii was found in the urine of one hare, which died on day 13 post infection (p.i.). Antibodies against C. burnetii persisted for one year of observation period, those against R. slovaca up to the 181st day p.i. Based on the results obtained and data on other lagomorph species, the hare could play an important role as host of C. burnetii and R. slovaca in nature.

Animals↗

Coxiella burnetii pericarditis: report of 15 cases and review.

Q fever is characterized by its clinical polymorphism, and pericarditis associated with Q fever has occasionally been described. Herein we report 15 cases of Coxiella burnetii pericarditis, 9 from our data bank and 6 encountered within the past 12 months. Three patients presented with life-threatening tamponade. We compare our cases with the 18 previously reported and with 60 Q fever-matched controls at our center. This study showed that Q fever pericarditis can present as acute as well as chronic disease; we describe relapse after 6 months in association with a serological profile compatible with the chronic form of disease (phase I C. burnetii IgG titer of > or = 800). Discriminant factors among patients and controls are age of > 52 years (adjusted odds ratio [OR], 5.66), the occurrence of general symptoms such as arthralgias or myalgias (adjusted OR, 6.54), and a normal erythrocyte sedimentation rate (adjusted OR, 16.37). No specific symptoms or underlying cardiac predispositions are observed.

Adolescent↗

[Prolonged febrile syndrome as a manifestation of Coxiella burnetii infection, presentation of infrequent clinical cases].

We present two cases of infection by Coxiella burnetii which developed with sustained fever symptoms. During its evolution, the first case presented granulomatous hepatitis, whereas the second case presented left Cosofemoral Arthritis. We describe the clinical-evolutive characteristics of these clinical forms and within the evolution of the chronic forms of Q fever.

Adult↗

Maturation of the Coxiella burnetii parasitophorous vacuole requires bacterial protein synthesis but not replication.

This study examined whether protein synthesis and replication are required for maturation and fusogenicity of the lysosomal-like, large and spacious parasitophorous vacuole (PV) of Coxiella burnetii, an obligate intracellular bacterium. Large and spacious PV with multiple non-replicating C. burnetii were observed by phase microscopy in Vero cells infected at a multiplicity of infection of ten and treated with a bacteriostatic concentration of nalidixic acid or carbenicillin, antimicrobics that inhibit DNA and cell wall biosynthesis respectively. Conversely, large and spacious PV were not observed in cells treated with a bacteriostatic concentration of the protein synthesis inhibitor chloramphenicol. Rather, fluorescence microscopy of individual cells revealed multiple, acidic PV harbouring a single organism tightly bounded by a LAMP-1 positive vacuolar membrane. These vacuoles homotypically fused to form a large and spacious PV upon removal of the drug. Chloramphenicol also inhibited trafficking of latex beads to large and spacious PV and caused mature PV to collapse. Collectively, these results demonstrate that C. burnetii protein synthesis, but not replication, is required for fusion between nascent C. burnetii PV and latex bead phagosomes, and also for formation and maintenance of large and spacious, replicative PV. However, transit of nascent PV through the endocytic pathway to ultimately acquire lysosomal markers appears to occur irrespective of Coxiella protein synthesis.

Animals↗

Coxiella burnetii seropositivity in parturient women is associated with adverse pregnancy outcomes.

OBJECTIVE: We conducted a cohort study of parturient women in an area with endemic Q fever infection to determine whether those seropositive for Coxiella burnetii had evidence of adverse birth outcomes. STUDY DESIGN: From June 1997 to November 1998, the cord blood of all women delivered at our health center was tested for antibodies to C burnetii by indirect immunofluorescence antibody test by using purified whole cell strain Nine Mile antigens. A titer of 1:8 or greater to either phase I or phase II antigens was considered seropositive. Placentas of a sample of cases and seronegative controls had polymerase chain reaction and culture performed. RESULTS: Evidence of prior infection with C burnetii was found in 3.8% (291/7658) of all parturient women. In a multivariate logistic regression, an association was seen between seropositivity (phase I titer >or= 1:8 or phase II titer >or= 1:32) and newborn gestational age >or=36 weeks (phase I antibody, odds ratio [OR] 2.4, 95% CI 1.3-4.3, P =.005; phase II antibody, OR 1.9, 95% CI 1.02-3.7, P =.04). Women with phase I antibody were more likely to have a prior or current neonatal death (phase I OR 3.2, 95% CI 1.09-9.3, P =.03). No placental samples from 153 seropositive or 93 seronegative women had Q fever by polymerase chain reaction or culture. CONCLUSION: About 4% of parturient women in this endemic area have evidence of previous exposure to C burnetii and this exposure is associated with adverse pregnancy outcomes. The pathogenesis of this association remains to be determined.

Adolescent↗

Failure of Q fever phase I corpuscular vaccine to influence the persistence and reactivation of Coxiella burnetii infection in mouse and guinea pig tissues.

In mice and guinea pigs infected with Coxiella burnetii, accumulation of large numbers of rickettsiae in the spleen and liver occurring at the early stages of infection was followed by clearing of these organs from the infectious agent but by its persistence in kidneys and reproductive tract at later intervals. The persistence of C. burnetii was not affected when one to six months-infected mice or guinea pigs were immunized with 100 or 500 micrograms of Q fever phase I corpuscular vaccine. Administration of the vaccine also did not substantially influence the reactivation of C. burnetii infection elicited in mice by parturition or developing in mice and guinea pigs upon treatment with cortisone or cyclophosphamide (CPA). Some differences observed between groups of immunized or non-immunized animals were only quantitative in nature. The possibility to disbalance the steady state of persisting C. burnetii in the host tissues by inhibition or stimulation of the immune response is discussed.

Animals↗

Roa307, a protein encoded on Coxiella burnetii plasmid QpH1, shows homology to proteins encoded in the replication origin region of bacterial chromosomes.

TnphoA mutagenesis identified an open reading frame, roa307, immediately upstream of the partition locus qsopAB on the Coxiella burnetii plasmid QpH1. The protein sequence deduced from roa307 displayed homology to Orf290 of Pseudomonas putida, Orf283 and Orf282 (SpoOJ) of Bacillus subtilis-hypothetical products of genes in the chromosomal replication origin region. Expression of roa307 was demonstrated by PhoA activity of an Roa307-PhoA fusion.

Amino Acid Sequence↗

Isolation of a Coxiella burnetii strain that has low virulence for mice from a patient with acute Q fever.

Coxiella burnetii was isolated from a patient with Q fever. It could not be determined whether this was an imported case or an indigenous one. Identification of the isolate was made by electron microscopic morphology and the indirect fluorescent antibody test with convalescent-phase serum from a Q fever patient having a known titer of antibody to C. burnetii. The isolated strain, named TK-1, caused no symptoms in ddY and BALB/c mice except when the mice were treated with cyclophosphamide.

Animals↗

Hemolytic-uremic syndrome associated with Coxiella burnetii infection.

Acute Q fever most commonly manifests as a self-limited febrile illness, pneumonia or hepatitis. We report the case of a 12-year-old child with documented exposure to sources of Coxiella burnetii who was admitted to our hospital because of hemolytic-uremic syndrome. Serologic tests established the diagnosis of acute Q fever.

Child↗

[Coxiella burnetii infections (Q fever) in animals and humans in Poznan and Leszno districts detected by serodiagnosis].

In Poznań and Leszno districts, serological survey of Q fever among cattle and sheep have been performed since 1973. Coxiella burnetii infections have not been detected until 1985. During this period 28,272 blood samples derived from cattle and sheep have been tested. For the first time in Poznań district, seropositive cattle were found in 1986 (5.5%). In the next year percentage increased to 16.3% and in following years has been on similar level. In Leszno district, serum antibodies to C. burnetii were detected in cattle in 1988 (25.9% seropositives). On the consecutive years, percentage of seropositive animals has decreased and in 1991 was 3.2% only. During these years there were 2.3 and 8.3% seropositive sheep detected in Poznań and Leszno districts respectively. Serological survey of C. burnetii infection has been performed among people exposed to this agent occupationally who undergo intermittent obligatory testing for brucellosis. In Leszno district among 4214 persons tested, 34.4% were seropositive. In Poznań district from 1988 to 1991 antibodies to C. burnetii have been detected in 22.7% of 6396 people tested. From year to year the percentage of seropositives fluctuated from 16.6 to 34.7%.

Animals↗

Immunochemical and antigenic characterization of Coxiella burnetii strains isolated in Europe and Mongolia.

SDS-PAGE, immunoblotting and serological methods such as microimmunofluorescence (MIF) test and ELISA were used to compare protein and lipopolysaccharide (LPS) profiles and antigenicity of 12 Coxiella burnetii strains isolated mostly from ticks in Europe and Mongolia with three reference C. burnetii strains originating from USA, namely Nine Mile from tick, Priscilla from goat placenta and S from human heart valve. Among strains from Europe and Mongolia, no significant differences in protein and LPS profiles were observed, irrespective of their origin, i.e. the country and source of isolation. The LPS profiles of these strains appeared to be more related to those of Nine Mile strain associated with acute Q fever, than to those of strains S and Priscilla associated with chronic Q fever. In immunoblots all strains isolated from Slovakia and Poland reacted more expressively with rabbit serum against Nine Mile than with serum against Priscilla strain. In the MIF test and ELISA there were no substantial differences in antibody-binding capacity between the reference and newly isolated C. burnetii strains, except for strain Priscilla reacting with homologous serum in lower antigenic concentration than other strains under study. However, in the MIF test much higher antigenic concentrations of each C. burnetii strain was required to detect antibodies in the Priscilla serum than in the Nine Mile, Luga and S sera.

Animals↗

[Detection of Coxiella burnetii in the air of a sheep barn during shearing].

Local epidemics of Q fever occur sporadically in Germany, mainly in rural residential communities. There is increasing evidence that these outbreaks, which are caused by Coxiella burnetii, are related particularly to the lambing season and shearing periods of nearby sheep holdings. It is assumed that this zoonotic agent is massively emitted from the placenta of infected ewes at birth and during shearing of wool contaminated with infected faeces of ticks. However, little is known about the airborne transmission and travel distance of this infectious agent, and only few attempts have been made to isolate it directly from the air. This paper describes for the first time the isolation and detection of C. burnetii in the air of an enclosed sheep barn during shearing of a herd which had tested positive for C. burnetii serologically and by PCR. Samples of inhalable dust samples were taken using I.O.M. samplers with glass fibre and polycarbonate filters at a flow rate of 2.5 l/min. The sampling time was nearly 4.5 h. Two sampling positions were set up on both sides of the shearing place at a distance of 3 m and 1.5 m above the ground. A third position with the same sampling equipment was not activated and served as a sampling and transport control. In the laboratory, the glass fibre filters were used to determine the dust concentration. The polycarbonate filters were treated in a specific breakdown procedure which inactivates PCR inhibitors, followed by amplification and sequencing of a specific DNA section of C. burnetii, which was found in the dust from both active sampling positions. The investigation clearly shows that the sampling and detection methods used in this small field study are suitable for the detection of C. burnetii in the air of sheep barns. The results confirm experimentally the high risk of airborne transmission of C. burnetii from sero-positive sheep herds during shearing. However, little is known about the effective travel distance of infective airborne C. burnetii particles. There is an urgent need for more detailed investigations on the emission and airborne dispersion of infectious C. burnetii particles in order to improve our understanding of the health risks caused by this zoonotic agent originating from sheep herds.

Air Microbiology↗

Detection of Coxiella burnetii specific DNA in blood samples from Japanese patients with chronic nonspecific symptoms by nested polymerase chain reaction.

The nested polymerase chain reaction (PCR) was used for direct species-specific detection of Coxiella burnetii in blood samples from 52 patients with chronic nonspecific symptoms, but no diagnostic or treatment history of Q fever. All patients had been in ill-health with general fatigue, muscle and joint pain, headache, etc., for one to more than 10 years. Seventeen (33%) showed evidence of C. burnetii infection, based on amplification of 438-bp fragments specific to C. burnetii by nested PCR, and 94% of positive patients reported close contact with animals. In contrast, five (9.6%) of 52 samples from healthy adult controls and two (2.8%) of 70 cord blood samples were positive by nested PCR. These data suggest a high prevalence of infection among adult patients with long term, nonspecific complaints who live in close contact with animals and the possible existence of a chronic post-acute Q fever syndrome in Japan.

Adolescent↗

Mononeuritis multiplex caused by Coxiella burnetii infection (Q fever).

After 1 week of flu-like illness, a 64-year-old man developed rapidly progressive mononeuritis multiplex involving the right arm and both legs. Serologic studies identified Coxiella burnetii as the cause of the febrile disease (Q fever). Fourteen days doxycycline treatment (200 mg daily) induced rapid and complete recovery. After 6 months, flu-like symptoms, weakness and hypalgesia of the right leg reappeared. Antibody titers again identified Q fever. Doxycycline was re-established and induced prompt recovery. Q fever has been associated with various neurologic complications such as meningoencephalitis, cerebellitis, optic neuritis or polyneuroradiculitis. This is the first report on Q fever related mononeuritis multiplex. Prolonged antibiotic treatment may be required to prevent relapsing infection from the resistant bacterium.

Anti-Bacterial Agents↗

Bactericidal effect of doxycycline associated with lysosomotropic agents on Coxiella burnetii in P388D1 cells.

There is no consistently reliable treatment for endocarditis resulting from chronic Coxiella burnetii infection, the causative agent of Q fever. Although certain antibiotics are recommended on the basis of their in vitro bactericidal activities, results of therapy with these antibiotics are often disappointing. To evaluate whether the currently recommended antibiotic susceptibility tests for C. burnetii give misleading results because of continued division of uninfected cells, thereby resulting in the dilution of infected cells and, hence, a false picture of antibiotic efficacy, we blocked cell division during antibiotic susceptibility testing with cycloheximide. Using this new method, we found that the currently recommended antibiotics for the treatment of Q fever, doxycycline, pefloxacin, and rifampin, did not reduce the ratio of infected to noninfected cells (either L929 or P388D1) by 9 days postinfection. To test the hypothesis that this lack of antibacterial activity is due to antibiotic inactivation by the low pH of the phagolysosomes in which C. burnetii is found, we used alkalinizing lysosomotropic agents (chloroquine or amantadine) concurrently with doxycycline. This resulted in the sterilization of C. burnetii infection in P388D1 cells. This finding seems to confirm our suspicion that the acidic conditions of the phagolysosomes in which C. burnetii is located inhibit antibiotic activity. This inhibition can be reversed in vitro when lysosomotropic alkalinizing agents are used.

Amantadine↗

[Time course of the levels of antibodies to Coxiella burnetii and detection of C. burnetii-DNA in three imported cases of acute Q fever].

Three patients developed acute Q fever after returning from an inspection tour of farms and abattoirs to Australia. Serum levels of antibodies to Coxiella burnetii and the presence of C. burnetii-DNA in blood samples were examined for more than 100 days. Four-fold raises of IgM and IgG antibodies against C. burnetii phase 2 were observed within the first three weeks in all the three cases. Maximum titers of IgM and IgG antibodies were 1,024-2,048 and 512-4,096, respectively. According to the temporal diagnostic criteria of acute Q fever in the convalescent serum: the IgM titer of > or = 64 and IgG titer of > or = 512 against phase 2, patient A, B and C were determined to be antibody positive for 45, 199 and 122 days, respectively. The result suggests that this standard is practical and reasonable for diagnosis of acute Q fever. C. burnetti-DNA was detected in the sera and buffy coat samples of patient A who developed high fever, severe thrombocytopenia and liver disfunction, but not in those of patient B and C. This study provides useful information for optimization and standardization of Q fever diagnosis in Japan.

Acute Disease↗

Transmission of antibodies to Chlamydia psittaci and Coxiella burnetii through eggs and "crop milk" in pigeons.

Young semi-domesticated pigeons captured or hatched from eggs gathered in Bratislava during 1989-1991 were examined for complement fixing antibodies to Chlamydia psittaci and agglutinating antibodies to Coxiella burnetii. Antibodies to Ch. psittaci were present in 76% of birds younger than 24 h, in 47.7% between 1 and 10 days of age and in 12% of nestlings over 10 days old. Antibodies to Ch. psittaci were also detected in "crop milk" of 4.1% of 1 to 10 day old birds and in 4.5% of specimens older than 10 days. Antibodies to C. burnetii were not found in juvenile birds under 24 h old, but antibodies against this agent were present in 16.4% birds between 1 and 10 days old and in 18% over 10 days old. Antibodies to C. burnetii were also detected in "crop milk" collected from crops of 2% of the young birds between 1 and 10 days.

Animals↗

Constrictive pericarditis due to Coxiella burnetii.

Q fever is characterized by its clinical polymorphism. Cardiac involvement in acute Q fever is rare. We report a case of pleuro-pericarditis that rapidly evolved to pericardial constriction during an acute episode of Coxiella burnetii infection. Constrictive pericarditis was confirmed by hemodynamic measurements, echocardiography, and magnetic resonance. Indirect immunofluorescence assay revealed positive serology for acute Q fever. The patient underwent a successful pericardiectomy and was given antibiotics. The histopathologic study of the excised pericardium showed C. burnetii in a large cluster of organisms. After a 6-month follow-up period, the individual was asymptomatic.

Antibodies, Bacterial↗