Burnetii Infection Research Articles

High Concentrations of Serum Soluble E-Cadherin in Patients With Q Fever.

Cadherins switching is a hallmark of neoplasic processes. The E-cadherin (E-cad) subtype is one of the surface molecules regulating cell-to-cell adhesion. After its cleavage by sheddases, a soluble fragment (sE-cad) is released that has been identified as a pro-carcinogenic inflammatory signal in several bacteria-induced cancers. Recently we reported that Q fever, a disease due to Coxiella burnetii infection, can be complicated by occurrence of non-Hodgkin lymphoma (NHL). Therefore, we studied E-cad switching in Q fever. The sE-cad levels were found increased in the sera of acute and persistent Q fever patients, whereas they remained at the baseline in controls groups of healthy donors, people cured of Q fever, patients suffering from unrelated inflammatory diseases, and past Q fever patients who developed NHL. These results indicate that sE-cad can be considered as a new biomarker of C. burnetii infection rather than a marker of NHL-associated to Q fever. We wondered if changes in sE-cad reflected variations in the CDH1 gene transcription. The expression of E-cad mRNA and its intracellular ligand β-catenin was down-regulated in peripheral blood mononuclear cells (PBMCs) of patients with either acute or persistent forms of Q fever. Indeed, a lower cell-surface expression of E-cad was measured in a minority (<5%) subpopulation of HLADR+/CD16+ monocytes from patients with acute Q fever. However, a very strong increase in E-cad expression was observed on more than 30% of the HLADR+/CD16+ monocytes of persistent Q fever patients, a cell subpopulation known to be a target for C. burnetii in humans. An experimental in vitro infection of healthy donors' PBMCs with C. burnetii, was performed to directly evaluate the link between C. burnetii interaction with PBMCs and their E-cad expression. A significant increase in the percentage of HLADR+/CD16+ monocytes expressing E-cad was measured after PBMCs had been incubated for 8 h with C. burnetii Nine Mile strain. Altogether, these data demonstrate that C. burnetii severely impairs the E-cad expression in circulating cells of Q fever patients.

Expression of human TLR4/myeloid differentiation factor 2 directs an early innate immune response associated with modest increases in bacterial burden during Coxiella burnetii infection.

Human TLR4 (hTLR4) and mouse TLR4 molecules respond differently to hypo-acylated LPS. The LPS of Coxiella burnetii is hypo-acylated and heavily glycosylated and causes a minimal response by human cells. Thus, we hypothesized that mice expressing hTLR4 molecules would be more susceptible to C. burnetii infection. Our results show that transgenic mice expressing hTLR4 and the human myeloid differentiation factor 2 (MD-2) adaptor protein (hTLR4/MD-2) respond similarly to wild type mice with respect to overall disease course. However, differences in bacterial burdens in tissues were noted, and lung pathology was increased in hTLR4/MD2 compared to wild type mice. Surprisingly, bone marrow chimera experiments indicated that hTLR4/MD-2 expression on non-hematopoietic cells, rather than the target cells for C. burnetii infection, accounted for increased bacterial burden. Early during infection, cytokines involved in myeloid cell recruitment were detected in the plasma of hTLR4/MD2 mice but not wild type mice. This restricted cytokine response was accompanied by neutrophil recruitment to the lung in hTLR4/MD2 mice. These data suggest that hTLR4/MD-2 alters early responses during C. burnetii infection. These early responses are precursors to later increased bacterial burdens and exacerbated pathology in the lung. Our data suggest an unexpected role for hTLR4/MD-2 in non-hematopoietic cells during C. burnetii infection.

A transcriptional signature associated with non-Hodgkin lymphoma in the blood of patients with Q fever.

Coxiella burnetii, the agent causing Q fever, has been associated with B-cell non-Hodgkin lymphoma (NHL). To better clarify this link, we analysed the genetic transcriptomic profile of peripheral blood leukocytes from patients with C. burnetii infection to identify possible links to lymphoma. Microarray analyses revealed that 1189 genes were expressed differently (p <.001 and fold change ≥4) in whole blood of patients with C. burnetii infection compared to controls. In addition, 95 genes expressed in patients with non-Hodgkin lymphoma (NHL) and in patients with C. burnetii persistent infection have allowed us to establish the ‘C. burnetii-associated NHL signature’. Among these, 33 genes previously found modulated in C. burnetii-associated -NHL by the microarray analysis were selected and their mRNA expression levels were measured in distinct C. burnetii-induced pathologies, namely, acute Q fever, focalized persistent infection, lymphadenitis and C.burnetii-associated NHL. Specific genes involved in anti-apoptotic process were found highly expressed in leukocytes from patients with C. burnetii associated-NHL: MIR17HG, REL and SP100. This signature differed from that found for NHL-control group. Patients with C. burnetii lymphadenitis presented significant elevated levels of BCL2 and ETS1 mRNAs. Altogether, we identified a specific transcriptionnal signature for NHL during C. burnetii infection reflecting the up-regulation of anti-apoptotic processes and the fact that lymphadenitis might constitute a critical step towards lymphomagenesis.

Bovine abortion caused by Coxiella burnetii: report of a cluster of cases in Uruguay and review of the literature.

A cluster of 4 bovine abortions caused by Coxiella burnetii occurred in a dairy herd in Uruguay during a 2-mo period. Case 1 consisted of a placenta from an aborted cow; cases 2-4 were fetuses and their placentas. Grossly, the placenta from one aborted cow had moderate, diffuse reddening of the cotyledons and loss of translucency of the intercotyledonary areas. No gross lesions were observed in the other 3 placentas. Microscopically, 2 of 4 placentas had fibrinonecrotizing placentitis with abundant intratrophoblastic gram-negative coccobacilli. C. burnetii was identified intralesionally by immunohistochemistry (IHC) in all 4 placentas, and by PCR and DNA sequencing in 3 placentas analyzed by these techniques. One fetus had mild neutrophilic alveolitis with multinucleate syncytial cells; no gross or microscopic lesions were observed in the other 2 fetuses examined. The lungs of the 3 fetuses were negative for C. burnetii by IHC. Tests performed to investigate other possible causes of abortions in the 4 cases were negative. C. burnetii causes Q fever in humans and coxiellosis in animals. Clusters of abortions in cattle by C. burnetii have not been reported previously, to our knowledge; this bacterium has been considered an opportunistic pathogen associated only with sporadic abortion in cattle. We present herein a cluster of 4 bovine abortions caused by C. burnetii in a dairy farm during a period of 2 mo and a review of the literature on C. burnetii infection in cattle.

Prevalence of Coxiella burnetii in Traditional and Industrial Butter and Cream Using Nested Polymerase Chain Reaction in Shahrekord, Iran

Background: A zoonotic disease of global importance known as "Q fever" is caused by Coxiella burnetii agent. The aim of the present research was to determine C. burnetii in the traditional and industrial butter and cream samples in Shahrekord, Iran. Methods: This cross-sectional descriptive study was conducted from March to September 2016. Totally, 200 traditional and industrial butter and cream samples were collected from retailers in different regions of Shahrekord and then tested for C. burnetii via nested polymerase chain reaction (PCR) assays. Results: In this survey, a total of 6 out of 200 samples (3%) were found to be positive for C. burnetii. More precisely, 4 (5.79%), 1 (5%), and 1 (2.56%) samples were related to the traditional bovine cream, traditional sheep butter, and traditional bovine butter samples, respectively. Nevertheless, no C. burnetii infections were found in the industrial butter and cream samples. Conclusions: These results proved that traditional dairy products can be considered as an important reservoir for C. burnetii infection in Iran.

Prevalence ofCoxiella burnetiiin Traditional and Industrial Butter and Cream Using Nested Polymerase Chain Reaction in Shahrekord, Iran

Background: A zoonotic disease of global importance known as "Q fever" is caused by Coxiella burnetii agent. The aim of the present research was to determine C. burnetii in the traditional and industrial butter and cream samples in Shahrekord, Iran. Methods: This cross-sectional descriptive study was conducted from March to September 2016. Totally, 200 traditional and industrial butter and cream samples were collected from retailers in different regions of Shahrekord and then tested for C. burnetii via nested polymerase chain reaction (PCR) assays. Results: In this survey, a total of 6 out of 200 samples (3%) were found to be positive for C. burnetii. More precisely, 4 (5.79%), 1 (5%), and 1 (2.56%) samples were related to the traditional bovine cream, traditional sheep butter, and traditional bovine butter samples, respectively. Nevertheless, no C. burnetii infections were found in the industrial butter and cream samples. Conclusions: These results proved that traditional dairy products can be considered as an important reservoir for C. burnetii infection in Iran.

Pitfalls of molecular diagnostic testing for Coxiella burnetii DNA on throat swabs

IntroductionCoxiella burnetii, the causative pathogen of Q fever, is regularly detected in throat swabs from patients without serological evidence of Q fever infection. C. burnetii is also frequently found in bulk tank milk from dairy cows. We evaluated the false positivity rate of polymerase chain reaction (PCR) for C. burnetii DNA on throat swabs and investigated whether recent consumption of C. burnetii DNA-positive cow milk could contribute to this phenomenon. MethodsC. burnetii PCR was performed on throat swabs obtained from patients in whom a throat swab was ordered for other diagnostic purposes; patients with community-acquired pneumonia (CAP); and healthy volunteers after consumption of commercial C. burnetii-containing cow milk products. ResultsC. burnetii DNA was found in 5.0% of throat swabs ordered for other diagnostic purposes and in 15.3% of throat swabs from CAP patients without serological evidence of Q fever pneumonia. The positive and negative predictive value of C. burnetii PCR on throat swabs for Q fever pneumonia were 66.7% (95% CI, 38.0–88.2) and 48.9% (95% CI, 41.3–54.6), respectively. After consumption of commercial C. burnetii-containing cow milk products, C. burnetii DNA could be detected in throat swabs for as long as 30 min after ingestion. ConclusionC. burnetii PCR on throat swabs is of low diagnostic value for Q fever pneumonia and was false positive in 15.3% of CAP patients without Q fever pneumonia. Recent consumption of C. burnetii-containing products can influence the outcome of C. burnetii PCR on throat swabs. Therefore, diagnosis of C. burnetii infection should be made in combination with serology or PCR performed on blood.

Molecular investigation of tick-borne infections in cattle from Xinjiang Uygur Autonomous Region, China

Tick-borne diseases cause significant losses to livestock production in tropical and subtropical regions. However, information about the tick-borne infections in cattle in Xinjiang Uygur Autonomous Region (XUAR), northwestern China, is scarce. In this study, nested polymerase chain reaction (PCR) assays and gene sequencing were used to detect and analyze epidemiological features of Babesia bovis, B. bigemina, Coxiella burnetii and Anaplasma bovis infections in XUAR. Out of 195 samples tested, 24 (12.3%), 67 (34.4%), 40 (20.5%) and 10 (5.1%) were positive for B. bovis, B. bigemina, C. burnetii and A. bovis, respectively. Sequencing analysis indicated that B. bovis SBP-4, B. bigemina Rap1a, C. burnetii htpB and A. bovis 16S rRNA genes from XUAR showed 99%–100% identity with documented isolates from other countries. Phylogenetic analyses revealed that B. bovis SBP-4, B. bigemina Rap1a, C. burnetii htpB and A. bovis 16S rRNA gene sequences clustered in the same clade with isolates from other countries. To the best of our knowledge, this is the first report of C. burnetii infection of cattle in XUAR. Furthermore, this study provides important data for understanding the distribution of tick-borne pathogens, and is expected to improve the approach for prevention and control of tick-borne diseases in China.

Expression of human TLR4/MD-2 directs an early innate immune response associated with modest increase in bacterial burden during Coxiella burnetii infection

Abstract Human and mouse TLR4 molecules respond differently to hypo-acylated LPS. The LPS of Coxiella burnetii is hypo-acylated, heavily glycosylated and known to cause a minimal response by human cells. We hypothesized that mice expressing hTLR4 molecules would be more susceptible to C. burnetii infection. Our results suggested that transgenic mice expressing hTLR4 and the human MD-2 adaptor protein (hTLR4/MD-2) generally respond similarly to wild type mice, contrary to our hypothesis. Small differences in bacterial burdens in spleens and/or lungs of infected mice were noted and lung pathology was increased in hTLR4/MD2 compared to wild type mice. Surprisingly, bone marrow chimera experiments indicated that hTLR4/MD-2 expression on non-hematopoietic cells, rather than the target cells for C. burnetii infection, accounted for increased bacterial burden. Very early during infection, cytokines involved in myeloid cell recruitment were detected in the plasma of hTLR4/MD2 mice, but not in wild type mice. This restricted cytokine response was accompanied by early neutrophil recruitment to the lung in hTLR4/MD2 mice. These data suggest that hTLR4/MD-2 may alter early responses to C. burnetii infection. Our results suggest that during C. burnetii infection, the species of origin for TLR4 may be less important than with other pathogens with hypoacylated LPS.

Bayesian estimation of sensitivity and specificity of a PCR method to detect Coxiella burnetii in milk and vaginal secretions in sheep and goat samples

Coxiella burnetii is a gram-negative and polymorphic rod bacterium that causes Q fever, a common zoonotic disease distributed worldwide. Widespread occurrences of the disease outbreaks indicate the importance of coordinated animal and human health efforts to control these outbreaks. Different tests are available to determine the C. burnetii infection status of a flock, but false negative responses may occur, as infectious animals can shed bacteria in milk intermittently, especially during an asymptomatic infection. In this study, a Bayesian latent class model was implemented to estimate the sensitivity (Se) and specificity (Sp) of a PCR method for the detection of C. burnetii in milk samples (PCRM) and vaginal swabs (PCRV) from Iranian sheep and goats. A nested PCR assay was conducted to detect infected animals among 170 milk samples and 170 vaginal swabs from goat flocks and 170 milk samples and 170 vaginal swabs from sheep flocks. We implemented a Bayesian latent class model to estimate the Se and Sp of a PCR method for the detection of C. burnetii in milk samples and vaginal swabs from sheep and goats. Estimations were based on the cross-classified results of PCRM and PCRV from the sheep and goat subpopulations. Positivity was 17.6 and 33.5%, respectively, for PCRM and PCRV samples among sheep. In goats, the apparent prevalence was 32.9 and 56.4% in PCRM and PCRV samples testing positive, respectively. This indicated the lower sensitivity of PCRM. The Se of PCRV was significantly higher than Se of PCRM, which corresponded to a higher rate of vaginal-positive, milk-negative PCR samples. In contrast, Sp of PCRV was lower than Sp of PCRM, representing the higher false-positive rate of vaginal swabs. The PCRV outperformed PCRM in terms of identifying latently infected sheep and goats; however, neither method could identify all latently infected sheep and goats, thus the combination is recommended to maximize our ability to identify infected animals. The true prevalence of C. burnetii infection was higher in Iranian goats than sheep.

Chemokine Receptor 7 Is Essential for Coxiella burnetii Whole-Cell Vaccine-Induced Cellular Immunity but Dispensable for Vaccine-Mediated Protective Immunity.

Protective immunity against Coxiella burnetii infection is conferred by vaccination with virulent (PI-WCV), but not avirulent (PII-WCV) whole-cell inactivated bacterium. The only well-characterized antigenic difference between virulent and avirulent C. burnetii is they have smooth and rough lipopolysaccharide (LPS), respectively. Mice were vaccinated with PI-WCV and PII-WCV. Humoral and cellular responses were evaluated using protein chip microarrays and ELISpots, respectively. Dendritic cell (DC) maturation after stimulation with PI-WVC and PII-WVC was evaluated using flow cytometry. Vaccine-challenge studies were performed to validate the importance of the receptor CCR7. Other than specific antibody response to PI-LPS, similar antibody profiles were observed but IgG titers were significantly higher after vaccination with PI-WCV. Furthermore, higher frequency of antigen-specific CD4+ T cells was detected in mice immunized with PI-WCV. PI-WCV-stimulated DCs displayed significantly higher levels of CCR7 and migratory ability to secondary lymphoid organs. Challenge-protection studies in wild-type and CCR7-deficient mice confirmed that CCR7 is critical for PI-WCV-induced cellular immunity. PI-WVC stimulates protective immunity to C. burnetii in mice through stimulation of migratory behavior in DCs for protective cellular immunity. Additionally, the humoral immune response to LPS is an important component of protective immunity.

The sexual dimorphism of anticardiolipin autoantibodies in acute Q fever patients

ObjectivesQ fever is a zoonotic disease caused by Coxiella burnetii which affects men more than women (sex ratio men/women: 2.2). Acute Q fever complications are associated with elevation of anticardiolipin (aCL) antibodies. Here, we investigate the sexual dimorphism of aCL antibodies during acute C. burnetii infection. MethodsIgG aCL antibodies were evaluated at the time of Q fever serological diagnosis with enzyme-linked immunosorbent assay. Results were analysed according to sex. ResultsAmong the 1323 patients with Q fever tested for aCL, 1013 had acute Q fever (692 men/321 women) and 310 had persistent focalized infection (226 men/84 women). In cases of acute Q fever, men presented a significantly higher proportion of positive aCL antibodies (351/692, 50.7%) than women (113/321, 35.2%) (p <0.05). In addition, men had significantly higher aCL antibodies levels than women (p <0.001). ConclusionsWe highlight a relationship between sex and markers of autoimmunity during Q fever. Further investigations are necessary to better understand the mechanisms of this sexual dimorphism.

Occurrence and risk factors of Coxiella burnetii in domestic ruminants in Lebanon

Coxiella burnetii causes diseases in humans (Q fever) and animals, domestic ruminants playing a major role in the epidemiology of the infection. Information on C. burnetii infection in Lebanon is scanty. In order to assess the prevalence of C. burnetii infection in ruminants, a cross-sectional study was undertaken in 2014. A total of 1633 sera from ruminants (865 cattle, 384 sheep and 384 goats) from 429 farms (173 cattle, 128 sheep and 128 goats), in seven provinces of Lebanon were randomly selected and assayed for the presence of antibodies.39.86% of farms (95% CI: 35.23–44.56) resulted positive. The seroprevalence was 30.63% in Cattle-farms, 46.88% in sheep-farms and 45.31% in goat-farms.Milk samples collected from 282 seropositive animals (86 cows, 93 sheep and 103 goats) from 171 positive farms were tested by a high sensitive Real-Time PCR targeted to the IS1111 transposon of C. burnetii. The overall prevalence in farms was estimated to be 14.04%. Cattle-, sheep- and goat farm prevalence rates were 15.09%, 10% and 17.24%, respectively.The findings of the study show that C. burnetii prevalence in Lebanese domestic ruminants is related to animal species and farming practices. Indeed, the mixed herds with sheep (p < 0.01), the presence of common lambing/kidding areas (p < 0.001) in farms where the use of disinfectants was not a routine practice (p < 0.05) were identified as important risk factors.The results of the study provide baseline information for setting up herd management and public health measures for the prevention and control of Q fever in Lebanon.

Promiscuous Coxiella burnetii CD4 Epitope Clusters Associated With Human Recall Responses Are Candidates for a Novel T-Cell Targeted Multi-Epitope Q Fever Vaccine.

Coxiella burnetii, the causative agent of Q fever, is a Gram-negative intracellular bacterium transmitted via aerosol. Regulatory approval of the Australian whole-cell vaccine Q-VAX® in the US and Europe is hindered by reactogenicity in previously exposed individuals. The aim of this study was to identify and rationally select C. burnetii epitopes for design of a safe, effective, and less reactogenic T-cell targeted human Q fever vaccine. Immunoinformatic methods were used to predict 65 HLA class I epitopes and 50 promiscuous HLA class II C. burnetii epitope clusters, which are conserved across strains of C. burnetii. HLA binding assays confirmed 89% of class I and 75% of class II predictions, and 11 HLA class II epitopes elicited IFNγ responses following heterologous DNA/DNA/peptide/peptide prime-boost immunizations of HLA-DR3 transgenic mice. Human immune responses to the predicted epitopes were characterized in individuals naturally exposed to C. burnetii during the 2007–2010 Dutch Q fever outbreak. Subjects were divided into three groups: controls with no immunological evidence of previous infection and individuals with responses to heat-killed C. burnetii in a whole blood IFNγ release assay (IGRA) who remained asymptomatic or who experienced clinical Q fever during the outbreak. Recall responses to C. burnetii epitopes were assessed by cultured IFNγ ELISpot. While HLA class I epitope responses were sparse in this cohort, we identified 21 HLA class II epitopes that recalled T-cell IFNγ responses in 10–28% of IGRA+ subjects. IGRA+ individuals with past asymptomatic and symptomatic C. burnetii infection showed a comparable response pattern and cumulative peptide response which correlated with IGRA responses. None of the peptides elicited reactogenicity in a C. burnetii exposure-primed guinea pig model. These data demonstrate that a substantial proportion of immunoinformatically identified HLA class II epitopes show long-lived immunoreactivity in naturally infected individuals, making them desirable candidates for a novel human multi-epitope Q fever vaccine.

Molecular prevalence of Coxiella burnetii in milk in Iran: a systematic review and meta-analysis

Q fever is a major zoonotic disease in the world. The aim of this meta-analysis was to estimate the prevalence of Coxiella burnetii in animal milk in Iran. We systematically reviewed the literature to identify eligible studies from January 2008 to June 2016 in English or Farsi (Persian) databases. We extracted the molecular prevalence of C. burnetii in milk from cows, goats, sheep, and camels in Iran. The total prevalence of C. burnetii in cow milk was 15.09% (95% CI 11.08-19.10) by PCR methods. The highest and lowest prevalence of Q fever agent were seen in the East Azerbaijan (25.55%) and Khorasan-Razavi (4.22%) provinces, respectively. The molecular prevalence of C. burnetii in goat milk was 7.80% (95% CI 3.54-12.07%). The provinces of Qom (0%) and Lorestan (44.71%) had the lowest and the highest frequency of C. burnetii infection in goat's milk, respectively. Total prevalence of C. burnetii in sheep milk was 3.79% (95% CI 0.72-6.87%). The highest frequency of C. burnetii in sheep milk was detected in the Khorasan-Razavi province (34.78%). The frequency of C. burnetii in camel milk was 1.43%. High infection of C. burnetii in milk is an important health problem in Iran, amplified by the traditional preparations of dairy products.

Genetic evidence of Coxiella burnetii infection in acute febrile illnesses in Iran.

Mounting evidence suggests that Q-fever is more prevalent in Iran than originally believed. However, in most parts of the country, clinicians do not pay enough attention to Q fever in their differential diagnosis. The aim of this study was to investigate the prevalence of Coxiella burnetii in suspected cases of acute Q fever in north-western Iran using molecular techniques. Febrile patients were enrolled in the study and investigated for C. burnetii infection. Sera samples were tested using real-time PCR for detection of IS1111 gene, and positive samples were confirmed with nested PCR. Nine patients (4.2%) out of 216 suspected cases were positive for C. burnetii. Weakness and fatigue, headache, and lethargy were the most prevalent clinical symptoms in acute Q fever patients. According to the results of this study and other reports of human cases in Iran, the diagnosis system of Q fever in Iran should be urgently revamped.

Molecular Detection of Coxiella burnetii in Cattle on Ulleung Island, Korea: A Population-based Study with Four Years of Follow Up

In a population-based study with 4 years of follow up, we evaluated the prevalence of Coxiella burnetii in cattle on Ulleung Island, Korea. In this study, the rates of C. burnetii infection in cattle on Ulleung Island were determined by PCR and were found to be 0.3–1.0% in the period 2011–2014. All 17 C. burnetii partial 16S rRNA gene sequences from PCR-positive cattle were identical and 2 geographic representatives were included in our analysis. The nucleotide sequences of the 2 samples showed high (98.4–100%) identity with C. burnetii sequences obtained from the GenBank. In this long-term tracking study, the number of cattle positive for C. burnetii on Ulleung Island was low. To prevent the transmission of C. burnetii on Ulleung Island, control strategy should include biosecurity improvement in surveillance, livestock management, administering suitable tests before purchasing animals to detect C. burnetii shedders, and restricting movements between herds.

Coxiella burnetii infection presenting as fever of unknown origin: a retrospective case series from Bulgaria

Purpose: Fever of unknown origin (FUO) is a perplexing medical problem. The causes for FUO are more than 200 diseases. The aim of the study was to present human clinical cases of Coxiella burnetii infection debuting as FUO. Methods & Materials: The following methods were conducted in the study: literature search, laboratory, imaging, and statistical methods. Criteria of David T. Durack and Alan C. Street were applied for FUO definition. For the etiological diagnosis indirect immunoenzyme assay (ELISA) for antibodies detection against Coxiella burnetii were used. Coxiella burnetii phase 1 IgA/IgG and Coxiella burnetii phase 2 IgG/IgM antibodies were detected in serum by indirect immunoenzyme assay (SERION ELISA classic, Virion/Serion, Würzburg, Germany). Results: For the period of January 2008 to March 2015 nine patients with FUO caused by Coxiella burnetii were hospitalized at the Department of Infectious Diseases, Military Medical Academy, Sofia (Bulgaria). Male gender were predominant (male/female – 77.8%/22.2%), mean age was 48.78 ± 14.52 years (range: 26–67), hospital stay was 9.78 ± 2.95 days (range: 5–15), fever duration was 54.33 ± 56.23 days (range: 21–180). Laboratory investigations estimated elevation of ESR 49.11 ± 31.74 mm/h (95% CI: 13.09–111.31), CRP 37.68 ± 37.62 mg/L (95% CI: 36.07–111.42) and Fibrinogen 5.69 ± 1.59 g/L (95% CI: 2.57–8.81). Mean values of liver enzymes were in reference range. Abdominal ultrasound and X-ray demonstrated 33.3% each of them Contribution to the final diagnosis. Transthoracic echocardiography found 22.2% Contribution. Serological methods presented 100% Contribution. Conclusion: Coxiella burnetii infection was accepted as a final diagnosis based on the integrated information from the applied methods. Active search and establishment of this pathogen in case of FUO should avoid potential complications and consequences in case of untreated Coxiella burnetii infection.

Genetic variations in innate immunity genes affect response to Coxiella burnetii and are associated with susceptibility to chronic Q fever

ObjectivesChronic Q fever is a persistent infection, mostly of aortic aneurysms, vascular prostheses or damaged heart valves, caused by the intracellular bacterium Coxiella burnetii. Only a fraction of C. burnetii-infected individuals at risk develop chronic Q fever. In these individuals, a defective innate immune response may contribute to the development of chronic Q fever. We assessed whether genetic variations in genes involved in the killing machinery for C. burnetii by macrophages, contribute to the progression to chronic Q fever. MethodsThe prevalence of 66 single nucleotide polymorphisms (SNPs) in 31 genes pivotal in phagolysosomal maturation, bacterial killing and autophagy, was determined in 173 chronic Q fever patients and 184 controls with risk factors for chronic Q fever and serological evidence of a C. burnetii infection. Associations were detected with univariate logistic regression models. To assess the effect of these SNPs on innate responses to C. burnetii, the C. burnetii-induced cytokine production and basal reactive oxygen species production of healthy volunteers was determined. ResultsRAB7A (rs13081864) and P2RX7 loss-of-function SNP (rs3751143) were more common in chronic Q fever patients than in controls. RAB5A (rs8682), P2RX7 gain-of-function SNP (rs1718119), MAP1LC3A (rs1040747) and ATG5 (rs2245214) were more common in controls. In healthy volunteers, RAB7A (rs13081864) and MAP1LC3A (rs1040747) influenced the C. burnetii-induced cytokine production. RAB7A (rs13081864) modulated basal reactive oxygen species production. ConclusionsRAB7A (rs13081864) and P2RX7 (rs3751143) are associated with the development of chronic Q fever, whereas RAB5A (rs8682), P2RX7 (rs1718119), MAP1LC3A (rs1040747) and ATG5 (rs2245214) may have protective effects.

Combined Laboratory Approach to Detection of Parvovirus B19 and Coxiella Burnetii in Patients with Fever of Unknown Origin

Fever of unknown origin (FUO) is one of the greatest challenges for clinicians and patients. There are more than 200 etiological agents of FUO, among these the most common is the role of infection, neoplasms, and diseases of connective tissue. The aim of the present study is to investigate the role of the infectious agents parvovirus B19 (B19V) and Coxiella burnetii (C. burnetii) in the development of fever of unknown origin by a set of immunoenzymatic and molecular methods. The present study included a total of 70 adult patients diagnosed with FUO and hospitalized in Bulgarian Hospitals. A control group of 26 healthy people were also included. Serological (indirect enzyme immunoassay test for detection of B19V and C. burnetii Ph. II specific IgM/IgG) and molecular (extraction and detection of infectious nucleic acids) methods were used. From all patients with FUO, a positive result for B19V-IgM was obtained in 18/70 (25.71%, 95% CI: 15.47 - 35.95) and the highest percentage was found in age groups 0 - 9 and 10 - 19 years. Protective B19V immunity and past viral infection was reported in 41/70 (58.57%, 95% CI: 47.03 - 70.11), and this percentage corresponded with the control group 16/26 (61.54%, 95% CI: 42.84 - 80.24). Anti-C. burnetii Ph. II-IgM was demonstrated in 13/70 (18.57%, 95% CI: 9.46 - 27.68). A relatively high percentage of affected patients were ≤ 40 years. Anti-C. burnetii Ph. II-IgG was detected in 24/70 (34.29%, 95% CI: 23.17 - 45.41). The control group has a 100% negative result for acute B19V and C. burnetii infection. A positive B19V-DNA result was obtained in 12/70 (17.14%, 95% CI: 8.31 - 25.97) patients. In 11/12 (91.67%) it was in combination with positive B19V-IgM marker. Of the total 70 sera tested, a positive PCR results for C. burnetii-DNA were obtained in 11 (15.71%, 95% CI: 7.18 - 24.24). According to clinical manifestation and concomitant symptoms, a high percentage of B19V and C. burnetii positives were associated with FUO and fever, headache, chills, and rash. It is of particular importance for a correct diagnosis of FUO to use a combined laboratory approach to prove acute or persistent infection and to test for a set of etiological agents.