Bartonella henselae is usually not viable in lymph nodes of patients with cat scratch disease.

Bartonella henselae, the agent of cat scratch disease (CSD), appears to be a common organism responsible for lymphadenitis in both adults and children. There is a very low isolation rate for B. henselae from lymph nodes of patients with CSD. Our objective was to evaluate B. henselae viability in a large series of lymph nodes from patients with CSD. From January to November 2016, we analyzed lymph node biopsy samples from patients diagnosed with CSD. We used reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to detect B. henselae RNA, as well as cultures, histological analyses, and fluorescence in situ hybridization (FISH). We tested 87 lymph nodes positive for B. henselae DNA but only 8 (9%) presented with B. henselae RNA. We did not find a significant difference for the pap threshold cycle (CT) values between RNA-positive and RNA-negative lymph nodes (p = 0.5). Cultures, histological analyses, and FISH were negative for all the tested samples. We provide evidence that B. henselae are not or are rarely viable in most cases in the lymph nodes of patients with CSD.

Cardiac magnetic resonance assessment of left ventricular dilatation in chronic severe left-sided regurgitations: comparison with standard echocardiography.

PURPOSE: The purpose of this study was to investigate the potential additional value of cardiac magnetic resonance (CMR) in the assessment of left ventricular (LV) dilatation and dysfunction by comparison to standard echocardiography in patients with chronic left-sided valvular regurgitation. MATERIALS AND METHODS: We prospectively enrolled patients with chronic severe mitral regurgitation (MR) or aortic regurgitation (AR). They underwent standard echocardiography and CMR using aortic flow and LV-function sequences. LV dilatation or dysfunction was assessed with each technique, based on thresholds used for surgery indication. Reference regurgitation severity was defined following previously reported CMR-based regurgitant volume thresholds. RESULTS: A total of 71 patients with chronic severe MR (n=44) or severe AR (n=27) were prospectively included. There were 60 men and 11 women with a mean age of 61±14 (SD) years (range: 18-83 years). CMR-based regurgitation severity was significantly greater in the LV dysfunction group when assessed with CMR (MR, P=0.011; AR, P=0.006) whereas it was not different when LV dysfunction was assessed using standard echocardiography. Among standard echocardiography and CMR volumetric indices, CMR-derived end-diastolic volume showed the best ability to predict regurgitation severity (area under the curve [AUC]=0.78 for MR; AUC=0.91 for AR). Diagnostic thresholds identified on receiver operating characteristics-curve analysis were lower than those of current European recommendations and closer to North-American guidelines. CONCLUSION: CMR assessment of LV end-diastolic volume in chronic severe left-sided regurgitations is more reliably associated with CMR-based regurgitant volume by comparison with standard echocardiography diameter. CMR may provide useful evaluation before surgery decision for severe asymptomatic regurgitations.

Epidemic of venereal treponematosis in wild monkeys: a paradigm for syphilis origin.

Treponema pallidum infections have been primarily known as slightly contagious mucocutaneous infections called yaws (tropical Africa and America) and bejel (subtropical North Africa). T. pallidum emerged as a highly infectious venereal syphilis agent in South America, probably about 500 years ago, and because of its venereal transmission, it quickly caused a worldwide pandemic. The disease manifests as lesions, including a chancre; then antibodies become detectable when or slightly after the chancre appears, and before the development of a rash and other systemic manifestations. Venereal diseases are poorly known in monkeys. During fieldwork in Senegal, we discovered an epizootic outbreak of venereal disease that we explored. We detected a venereal form of T. pallidum subsp. pertenue infection in green monkeys (Chlorocebus sabaeus), then observed an epizootic outbreak in Senegal and its spread among baboons a year later. Comparative analysis of T. pallidum genomes from the monkeys' chancres and other Treponema genomes showed an acceleration of the number of single nucleotide polymorphisms, comparable to that observed in syphilis. Identified T. pallidum clones seem to be epizootic through the acceleration of their mutation rate, which is linked to their larger diffusion.

A 2,000-year-old specimen with intraerythrocytic Bartonella quintana.

Photogrammetry and cascading microscopy investigations of dental pulp specimens collected from 2,000-year-old individuals buried in a Roman necropolis in Besançon, France, revealed unprecedented preserved tissular and cellular morphology. Photogrammetry yielded 3-D images of the smallest archaeological human remains ever recovered. Optical microscopy examinations after standard haematoxylin-phloxine-saffron staining and anti-glycophorin A immunohistochemistry exposed dental pulp cells, in addition erythrocytes were visualised by electron microscopy, which indicated the ancient dental pulp trapped a blood drop. Fluorescence in situ hybridisation applied on red blood cells revealed the louse-borne pathogen Bartonella quintana, a finding confirmed by polymerase chain reaction assays. Through paleohistology and paleocytology, we demonstrate that the ancient dental pulp preserved intact blood cells at the time of the individual's death, offering an unprecedented opportunity to engage in direct and indirect tests to diagnose pathogens in ancient buried individuals.

[Q fever: current diagnosis and treatment options]. / Fièvre Q: actualités diagnostiques et thérapeutiques.

Q fever is a zoonotic disease caused by the ubiquitous pathogen Coxiella burnetii responsible for acute and chronic clinical manifestations. Its geographically heterogeneous prevalence seems mainly related to the clinician interest and the availability of a reference center. Its polymorphic clinical expression imposes reference to diagnosis in presence of pneumonia, hepatitis, prolonged fever or endocarditis with no proof of its etiology. The diagnosis is mainly serological. If acute Q fever is most often benign, endocarditis is constantly fatal without treatment. The treatment is effective and well tolerated, but must be adapted to the acute or chronic pattern, the presence of a heart valve disease, an aneurysm or a vascular prosthesis, an immunodeficiency and the specific problem of pregnancy.

The hypervirulent Coxiella burnetii Guiana strain compared in silico, in vitro and in vivo to the Nine Mile and the German strain.

OBJECTIVE: Q fever epidemic outbreaks have been reported in French Guiana and in The Netherlands. To determine whether the C. burnetii strains involved in these epidemics had a peculiar virulence pattern, we compared the pathogenicity of the Guiana and the German strain (a clone of The Netherlands strain), in silico, in vitro, and in vivo versus the Nine Mile strain. METHOD: The pan-genomes of the Guiana (Cb175), German (Z3055), and the referent Nine Mile (RSA 493) C. burnetii strains were compared. In vitro, the growth rate and the morphological presentation were compared. In vivo (SCID and Balb/c mice), weight loss, histological lesions, C. burnetii bacterial load in deep organs, and serological response were reported according to each C. burnetii strain studied. RESULTS: The Guiana strain had 77 times more missing genes and 12 times more unique genes than the German strain. The Guiana strain presented as large cell variants (LCVs) and led to the most pronounced fatality rate in SCID mice (100% at 4 weeks). The German strain presented as small cell variants (SCVs), and had an intermediate fatality rate (75% at 4 weeks). Both the Guiana and the German strains led to a significant higher serological response at 2 and 4 weeks post infection (p <0.05). CONCLUSION: The Guiana strain was the most virulent strain, followed by the German strain and the referent Nine Mile strain. Unique and missing genes could be implicated but further investigations are necessary to specify their role.

A case of giant cell arteritis associated with culture-proven Coxiella burnetii aortitis.

A case of proven Coxiella burnetii aortitis, possibly associated with giant cell arteritis (GCA), is reported. A 72-year-old man, who is a hunter, presented with weight loss, fever, jaw claudication, and hardened temporal arteries associated with a persistent inflammatory syndrome and arteritis of the whole aorta, including the brachiocephalic arteries, as seen on 18F-fluorodeoxyglucose positron emission tomography/computed tomography. The diagnosis of GCA was retained, and treatment with prednisolone was started. Given the aneurysm of the abdominal aorta, the patient underwent replacement of the abdominal aorta with an allograft. Histology showed intense chronic arteritis attributed to atherosclerosis with dissection. However, Coxiella burnetii infection was confirmed by serology and then by culture and molecular biology on the surgical specimen. A combination of hydroxychloroquine and doxycycline was added to tapered prednisolone and the outcome was favourable.

Chronic nasal infection caused by Klebsiella rhinoscleromatis or Klebsiella ozaenae: two forgotten infectious diseases.

OBJECTIVES: Klebsiella rhinoscleromatis and Klebsiella ozaenae are associated with chronic diseases of the upper airways: rhinoscleroma and ozena, respectively. These have become uncommon in developed countries. We report herein one case of each disease in patients living in Marseilles, France, and include a review of the literature. METHODS: Diagnosis was made by direct evidence of bacteria (specific cultures and autoimmunohistochemistry on nasal biopsy) and using an indirect method (serology). In addition, the literature review showed that the majority of publications were old, confirming the fact that these diseases have been long forgotten. RESULTS: The specific and original methods used have allowed us to confirm the pathogenic role of K. ozaenae in ozena and confirmed rhinoscleroma in a granulomatous lesion. In the literature, K. rhinoscleromatis is only associated with rhinoscleroma whereas K. ozaenae is also associated with clinical diseases other than chronic rhinitis. CONCLUSIONS: In cases of chronic rhinitis, ozena and rhinoscleroma should be kept in mind, even in developed countries, and systematically screened for, especially as there are specific diagnostic tools and effective treatments available.

Growth-promoting effects of single-dose intragastrically administered probiotics in chickens.

1. Live body weight gain (WG) and food conversion efficiency (FCE) were measured over a 4-week period in chicks (female, Kabir strain), inoculated intragastrically at one week of age with either a single dose (1 ml) of 10(7) live bacterial cells of Lactobacillus fermentum (LBF) or a strain of Lactobacillus spp. (LB), named Autruche 4 (A4), suspended in 1 ml of phosphate-buffered saline (PBS, pH 7.0); controls (n = 10) were inoculated only with PBS. The same commercial chicken feed was used throughout the 4-week experiment. 2. Compared with the control group, WG to 8 and 21 d, and also liver weight, were significantly greater in LBF and A4 treatments and FCE improved in both of the experimental groups. Compared with the A4 treatment group, WG was significantly higher and FCE improved in the LBF-treated group. The effects of A4 treatment on chicks were similar to those in an earlier pilot experiment using the same methodology but with 1-week-old female chicks (Kabir) inoculated with either the A4 strain or strain of LB named Autruche 5. 3. It is concluded that a single dose of Lactobacillus fermentum or Lactobacillus spp. (Autruche 4) administered intragastrically improves WG and FCE of broiler chicks.

Cytomegalovirus reactivation enhances the virulence of Staphylococcus aureus pneumonia in a mouse model.

OBJECTIVES: Cytomegalovirus (CMV) reactivation in intensive care unit patients may increase mortality and favour bacterial pneumonia. We developed a murine model to compare the severity of staphylococcal pneumonia after CMV reactivation and in CMV-negative mice. METHODS: Balb/c mice were primo-infected with murine cytomegalovirus (MCMV n=90) or received saline (control n=90). After latency, all mice underwent caecal ligation and puncture to trigger MCMV reactivation in MCMV primary-infected mice. Surviving animals received an intra-nasal inoculation with methicillin-susceptible Staphylococcus aureus (MSSA) to induce pneumonia. Mortality, lung bacterial count, histology and interferon-alpha and gamma serum levels were compared in MCMV reactivated and control mice 2, 5 and 15 days after pneumonia. RESULTS: After MSSA pneumonia, MCMV mice showed a trend towards a higher mortality (9.4% versus 0%; p 0.09) and a higher weight loss (2.2 (0.6-4.1 g) versus 0.7 (-0.3 to 1.3 g); p 0.005). The lung bacterial count was higher in MCMV mice 2 days (5×103 (103 to 3×105) versus 102 (0 to 4×102) CFU/lung; p 0.007) and 5 days (2.5×104 (1.6×104 to 6.5×105) versus 15 (10-40) CFU/lung; p 0.005) after MSSA pneumonia. 8/40 (20%) MCMV mice developed lung abscesses compared to 0% in control (p 0.011). Interferon-alpha serum levels 2 days after staphylococcal pneumonia were higher in MCMV mice. CONCLUSIONS: MCMV reactivation decreased lung bacterial clearance and favoured the development of staphylococcal abscessing pneumonia. CMV reactivation may be responsible for a higher susceptibility to bacterial sepsis.

Cerebral infarction: an unusual complication of Mediterranean spotted fever.

Mediterranean spotted fever is a rickettsiosis caused by Rickettsia conorii and transmitted by the brown dog tick. It is considered as a benign disease but 5% to 10% of patients present with a malignant form which is the result of a diffuse vasculitis. We report here the first case of Mediterranean spotted fever with cerebral vasculitis and thrombosis leading to a massive cerebral infarct and death.

Severe lower limbs lymphedema following breast carcinoma treatment revealing radiation-induced constrictive pericarditis--a case report.

In patients treated for breast carcinoma, unilateral lymphedema of the upper limb is usual. However, to the authors' knowledge, lower limb lymphedema has never been reported as a complication of breast carcinoma therapy. They report here the first case of a radiation-induced constrictive pericarditis revealed by severe lower limbs lymphedema. A 60-year-old woman was treated for left breast carcinoma with quadrantectomy, axillary lymphadenectomy, and combined radio chemotherapy (60 grays). Three and a half years later she suffered from a diffuse and increasing lower limbs lymphedema, which became huge and disabling. Radiation-induced constrictive pericarditis was evidenced by right cardiac cavities catheterization. A dramatic improvement was rapidly obtained after pericardectomy. Histopathologic analysis of the pericardium did not reveal neoplastic cells. Radiation-induced constrictive pericarditis is usually responsible for lower limbs edema, but lymphedema is exceptional. This case highlights the need to search for a constrictive pericarditis also in the case of lower limbs lymphedema, particularly in a patient treated with mediastinal radiotherapy or combined radio chemotherapy.

[Diagnosis of infectious lymphadenitis]. / Diagnostic des adénites infectieuses.

Adenitis is a common disorder requesting numerous medical specialties. Etiologies are dominated by viral and bacterial infections, and more rarely parasitic, or by neoplastic and inflammatory diseases. Nevertheless, etiology remains often unknown and invasive tests may be required. On nodal tissue sample, histological examination, culture and polymerase chain reaction (PCR) are realized. PCR has revolutionized the diagnostic approach and consequently, knowledge of infectious lymphadenopathy. Previously, staphylococcus, streptococcus and mycobacterium were the main infectious agents identified in lymph nodes. Since its use, new emergent microorganisms responsible of lymphadenitis have been identified. Bartonella henselae, responsible of cat scratch disease, is to date the infectious agent most often encountered in adenitis. Mycobacterium avium subsp. hominisuis has been recently described as responsible of children lymphadenitis. PCR has become an essential tool in the diagnostic process of infectious lymphadenitis. Here, we propose a literature review on infectious adenitis and we emphasize the diagnostic strategy of adenitis.

Bacillary angiomatosis in immunocompromised patients.

OBJECTIVES: To report seven cases of bacillary angiomatosis; to evaluate the most useful diagnostic tools; to analyse the clinical and epidemiological features associated with Bartonella quintana or Bartonella henselae infections. DESIGN: Clinical, diagnostic and epidemiological evaluation of 37 speciated bacillary angiomatosis cases in the literature, including the seven patients in our study. METHODS: Pathological examination of tissue samples, including Warthin-Starry staining and immunohistology; titre of antibodies to Bartonella sp.; detection of Bartonella sp. in blood and biopsy materials by culture or PCR; and statistical analysis of clinical and epidemiological features associated with B. quintana or B. henselae bacillary angiomatosis cases. RESULTS: Seven immunocompromised patients (six with AIDS and one patient with acute leukaemia) had bacillary angiomatosis confirmed by histology. B. quintana was cultured in three patients, whereas B. henselae DNA was amplified by PCR in the remaining four patients. Serum from only one patient reacted with Bartonella antigens. Amongst the 14 B. quintana and 23 B. henselae bacillary angiomatosis cases now reported in the literature, lymphadenopathies were significantly more frequent in B. henselae-infected patients, and neurological disorders of the central nervous system in B. quintana-infected patients. Risk factors were contact with cats, and homelessness or poor socioeconomic status in B. henselae and B. quintana bacillary angiomatosis cases, respectively. CONCLUSIONS: Although diagnosis of bacillary angiomatosis often remains solely based upon histology, culture or PCR-based methods are useful for the detection of Bartonella sp., and allow identification of the species involved, which is necessary to further characterize clinical and epidemiological features associated with B. quintana or B. henselae infections.

Quantitative analysis of valvular lesions during Bartonella endocarditis.

Cardiac valve pathology was evaluated in 15 patients with confirmed diagnosis of Bartonella endocarditis. Ten were infected by Bartonella quintana and 5 by Bartonella henselae. Histologic features of these cases, including fibrosis, calcification, vegetation, pattern of inflammation, and vascularization, were compared with those of valves from 25 cases of non-Bartonella endocarditis as controls using a computerized quantitative image analysis. Pathologic and immunohistologic testing for localization of Bartonella species in resected valves included Warthin-Starry stain and polyclonal antibody-based immunodetection. Compared with other cases of infective endocarditis, cases of Bartonella endocarditis are more fibrotic and calcified, less vascularized, with less extensive vegetation and chronic inflammation. These pathologic changes are suggestive of a prolonged infection. Warthin-Starry stain and immunohistologic testing demonstrated the presence of the organism, respectively, in 11 and 10 of the 13 tested valves. Results of both staining methods showed microorganisms in extracellular locations and in regions unaccompanied by inflammation. Pathology and immunohistology may contribute to the etiologic diagnosis of Bartonella endocarditis when serology and molecular techniques are not available.

Cat scratch disease with lymphadenitis, vertebral osteomyelitis, and spleen abscesses.

In this report we describe a 30-year old male patient with vertebral osteomyelitis and spleen abscesses with cat scratch disease. The diagnosis was made on the basis of molecular detection of Bartonella henselae either on lymph node biopsies or on bone biopsy, histology of the lymph node, serology using either our in-house microimmunofluorescence assay or a commercial kit (Focus Technologies). Immunofluorescent detection was also performed directly on slide appositions using a monoclonal antibody. Treatment consisted of administration of antibiotics with rapid clinical improvement and a stabilization of skeletal lesions on the magnetic resonance imaging performed three months later. Twenty two other cases of this unusual manifestation associated with cat scratch disease have been reported in the literature and are reviewed here. Our case represents the second case of osteomyelitis associated with cat scratch disease in which B. henselae has been specifically identified as the etiological agent using several direct and indirect methods.

Comparative pathology, and immunohistology associated with clinical illness after Ehrlichia phagocytophila-group infections.

The Ehrlichia phagocytophila-group also includes E. equi and the human granulocytic ehrlichiosis (HGE) agent that are probably a single species. Disease is mild to severe illness in ruminants, horses, and humans, but the comparative pathology and ehrlichial distribution in tissues is poorly described. We compared pathology and ehrlichial distribution in humans with HGE, horses with E. equi infection, and a sheep with E. phagocytophila infection. Frequent findings included splenic lymphoid depletion, small macrophage aggregates and apoptoses in liver, and paracortical hyperplasia in lymph nodes. Bone marrow was normocellular or hypercellular. Only the spleen was frequently infected; other organs with infected cells included lung, liver, heart, and kidney, but lesions were present in lung and liver only. Most infected cells were neutrophils. Ehrlichia phagocytophila-group infections are associated with moderate tissue damage. While the pathogenesis of granulocytic ehrlichiosis is not clear, pathologic studies suggest that the process is initiated by ehrlichia-infected cells but may result from host-mediated injury and immunosuppression.

Hidden chromosome abnormalities in a primary central nervous system lymphoma detected by multicolor spectral karyotyping.

Cytogenetic analysis provides important information for diagnosis and prognosis in some tumors. But karyotype analysis can be difficult in some cases, because metaphase chromosomes are contracted. New techniques, such as fluorescence in situ hybridization and, more recently, spectral karyotyping, or SKY, based on the hybridization of 24 fluorescently labeled chromosome painting probes, allow the detection and identification of complex chromosomal rearrangements. We report here a case of primary central nervous system lymphoma in which chromosomal rearrangements and marker chromosomes not identified by a routine cytogenetic technique were clarified by SKY. This shows the value of the SKY technique in the cytogenetic diagnosis of tumors.

Clinically and histologically silent Q fever endocarditis accidentally diagnosed by PCR.

A case of Q fever endocarditis was diagnosed in a patient with no sign of active endocarditis by performing PCR targeting eubacterial 16S rDNA on the resected mitral valve. The diagnosis was confirmed by detection of high levels of anti-Coxiella burnetti antibodies, positive immunohistologic analysis of the valve tissue with specific antibodies and culture of C. burnetti from the valve tissue. As this patient had an unexplained aggravation of valve dysfunction, we recommended routine serologic testing for C. burnetti to allow the diagnosis of Q fever endocarditis at a very early stage.

Expression of developmentally regulated cytoskeleton and cell surface proteins in childhood spinal muscular atrophies.

Expression of some developmentally regulated cytoskeleton components (desmin, vimentin and myosin heavy chain isoforms) and cell surface proteins (including neural cell adhesion molecule (NCAM), its polysialylated (PSA) isoform and CD24) have been studied by immunohistochemical detection in a series of 23 infantile spinal muscular atrophies (SMA). According to the clinical classification established by Byers and Banker in 1961, 8 cases were type I SMA (Werdnig-Hoffmann's disease), 10 cases were type II (intermediate form), and 5 cases were type III (Kugelberg-Welander's disease). In 15 cases, the percentage of immunoreactive fibers with the various antibodies used has been quantified and the results correlated with clinical data. The aim of the study was to search for variations in the pattern of expression of the proteins to improve the accuracy of diagnosis and prognosis, and to gain an understanding of the pathological processes involved in SMA. The results showed that the pattern of expression of these cytoskeleton and cell surface proteins is abnormal in all types of SMA. However, it was strikingly different in type I and II SMA as opposed to type III. In type I and II SMA, strong NCAM and developmental myosin heavy chain (MHC) expression was observed in atrophic fibers. Numerous atrophic fibers co-expressed desmin and vimentin as well as slow and fast adult MHC. Very few of them expressed PSA NCAM, fetal MHC and CD24. In type III SMA, the number of fibers expressing NCAM, developmental MHC and co-expressing slow and fast adult MHC was low and virtually none of them expressed vimentin or desmin. These findings are in favor of a denervation process occurring very early in life, probably even in utero, in type I and II SMA and leading to a severe impairment of muscle fibers maturation. In contrast, in type III SMA, the process is initiated well after birth and affects mature muscle fibers. In all types of SMA, the ability of muscle fibers to regenerate is low, although some fibers may be reinnervated. Immunohistochemical data was not related to the patients follow-up and thus has no prognostic value.