Spotted fever rickettsioses causing myocarditis and ARDS: a case from Sri Lanka.

BACKGROUND: Spotted fever group of rickettsial infections are emerging in Sri Lanka. We describe a patient with rapidly progressing ARDS and myocarditis secondary to spotted fever caused by Rickettsia conorii. ARDS and myocarditis are rare complications of Rickettsia conorii infections and only a few cases are reported to date. CASE PRESENTATION: A 53 years old manual worker presented with fever for 5 days and a skin rash. He was in circulatory failure on admission and developed severe hypoxaemia with gross changes in chest radiograph by next day requiring assisted ventilation. He had myocarditis causing left ventricular failure and acute respiratory distress syndrome. He was confirmed to have spotted fever rickettsial infection with rising titre of indirect immunofluorescence antibodies to Ricketssia conorii and made a complete recovery with appropriate antibiotic therapy and supportive care. CONCLUSION: Rickettsial infections can present with diverse manifestations. Even the patients with severe organ involvements such as myocarditis and ARDS can be completely cured if timely identified and treated.

[A Rickettsia case caused by Rickettsia conorii]. / Rickettsia conorii'nin neden oldugu riketsiyal enfeksiyon olgusu.

Rickettsia species are gram-negative intracellular, small pleomorphic coccobacilli in the Rickettsiaceae family. This genus is serologically and genotypically divided into four groups as spotted fever group, typhus group, Rickettsia belli and Rickettsia canadensis. Rickettsia conorii (R.conorii subsp. conorii) in the spotted fever group was reported to cause mediterranean spotted fever in Europe, especially in mediterranean countries including Turkey. The major vectors of Rickettsia species are ticks, and in some species fleas or mites. In this report a case with R.conorii infection was presented. A 46-year-old female patient, who had anorexia, fatigue, muscle aches, chills and high fever was admitted to a health institution. The patient was diagnosed as influenza. There was no regression in the patient's complaints with the recommended treatment. The patient was examined in our infectious diseases clinic and had several symptoms like severe muscle and joint pain with significant headache, and rashes at her body including hands and feet. The patient had a single eschar in the upper midline of the belly that matched tick biting and pink small maculopapular scars on the trunk, arms, legs, feet, and hands. Considering a Rickettsia pre-diagnosis, liquid electrolyte and doxycycline 2 x 100 mg oral treatment was started. On the third day of treatment, high fever, muscle and joint pain were decreased. On the fifth day, active skin lesions were started to fade. R.conorii IgM and IgG were negative in the first serum sample of the patient. In the biopsy sample taken from eschar tissue, Rickettsia spp. was detected as positive with rt-PCR. PCR was used by using the specific regions of the genetically specific gltA and ompA genes in the biopsy specimens and then the PCR products were determined by DNA sequence analysis. The DNA sequence results were compA red with Genbank data and determined that the gltA sequence was 99%, similar to R.conorii with accession number JN182786 and the ompA sequence was 99%, similar to R.conorii with accession number KR401144. When the phylogenetic tree was created, it was observed that the etiological agent was R.conorii. A week after the treatment, in the second serum sample R.conorii IFA IgM 1/192 titer and IgG 1/320 titer were detected as positive. In this case report, we have presented a Rickettsia case, clinically diagnosed as Rickettsia, serologically negative in the acute phase, PCR positive, with post-treatment seroconversion and etiologic agent determined as R.conorii.

Epidemiological evaluation of Mediterranean spotted fever in children of the Karak province in south Jordan.

INTRODUCTION: The aim of this study was to describe the epidemiological patterns of Mediterranean spotted fever (MSF) as well as its treatment and outcomes in children in south Jordan. METHODOLOGY: A retrospective observational study was conducted from June 2013 to December 2015. Data regarding demographics, clinical presentation, laboratory findings, treatment, and outcomes were collected. RESULTS: In total, 35 male and 20 female patients (mean age: 6 years ± 3.6) were included. The incidence of MSF was 7.9 cases/100,000 inhabitants/year; MSF affected 89% of individuals in the summer, 74.5% of those living in a rural area with tent housing, and 100% of those who had contact with animals. All cases presented with fever, and 94.5% had a skin rash. Serological tests were positive in 87.2% of cases, and Rickettsia conorii (the Moroccan strain) was present in all positive cases. All cases had thrombocytopenia, but none had leukocytosis. Hyponatremia was present in 71% of cases, and 49%, 61.8%, and 72.7% had increased urea, alanine transaminase, and aspartate aminotransferase levels, respectively. Doxycycline was administered to all patients, with a cure rate of 96.4% and mortality rate of 3.6%. CONCLUSIONS: MSF caused by R. conorii (the Moroccan strain) is prevalent in Jordan, and contact with animals is the route of transmission. The patients' responses to doxycycline were excellent. A high index of suspicion, early diagnosis, and specific treatment considerably decrease mortality. MSF should be considered as a possible cause of febrile disease in those with a rash and in those living in rural areas.

Scrub typhus and spotted fever among hospitalised children in South India: Clinical profile and serological epidemiology.

BACKGROUND: Rickettsial infections are re-emerging. In India, they are now being reported from several areas where they were previously unknown. OBJECTIVES: The objective of this study was to describe the epidemiology, clinical profile and outcome of serologically-confirmed scrub typhus and spotted fever among children in a tertiary care hospital in Bengaluru. MATERIALS AND METHODS: Hospitalised children aged <18 years, with clinical features suggestive of rickettsial disease admitted between January 2010 and October 2012 were included prospectively. Diagnosis was based on scrub typhus and spotted fever-specific IgM and IgG by enzyme-linked immunosorbent assay (ELISA). RESULTS: Of 103 children with clinical features suggestive of rickettsial illness, ELISA test confirmed 53 cases for scrub typhus, 23 cases for spotted fever group and 14 with mixed infection. The average age was 7.3 (±3.9) years and 44 (71.0%) children were male. Majority of cases were from Karnataka (50%), Andhra Pradesh (32.3%) and Tamil Nadu (17.7%). Common clinical features included fever (100%, average duration 11 days), nausea and vomiting (44%), rash (36%); eschar was rare. Compared to the ELISA test, Weil-Felix test (OX-K titre of 1:80) had a sensitivity and specificity of 88.7% and 43.9%, respectively. Treatment with chloramphenicol or doxycycline was given to the majority of the children. Complications included meningoencephalitis (28%), shock (10%), retinal vasculitis (10%) and purpura fulminans (7%). CONCLUSIONS: These findings suggest that the burden of rickettsial infection among children in India is high, with a substantially high complication rate. Rickettsial-specific ELISA tests can help in early diagnosis and early institution of appropriate treatment that may prevent life-threatening complications.

High serum CXCL10 in Rickettsia conorii infection is endothelial cell mediated subsequent to whole blood activation.

BACKGROUND: The pathophysiological hallmark of Rickettsia conorii (R. conorii) infection comprises infection of endothelial cells with perivascular infiltration of T-cells and macrophages. Although interferon (IFN)-γ-induced protein 10 (IP-10)/CXCL10 is induced during vascular inflammation, data on CXCL10 in R. conorii infection is scarce. METHODS: Serum CXCL10 was analyzed in two cohorts of southern European patients with R. conorii infection using multiplex cytokine assays. The mechanism of R. conorii-induced CXCL10 release was examined ex vivo using human whole blood interacting with endothelial cells. RESULTS: (i) At admission, R. conorii infected patients had excessively increased CXCL10 levels, similar in the Italian (n=32, ∼56-fold increase vs controls) and the Spanish cohort (n=38, ∼68-fold increase vs controls), followed by a marked decrease after recovery. The massive CXCL10 increase was selective since it was not accompanied with similar changes in other cytokines. (ii) Heat-inactivated R. conorii induced a marked CXCL10 increase when whole blood and endothelial cells were co-cultured. Even plasma obtained from R. conorii-exposed whole blood induced a marked CXCL10 release from endothelial cells, comparable to the levels found in serum of R. conorii-infected patients. Bacteria alone did not induce CXCL10 production in endothelial cells, macrophages or smooth muscle cells. CONCLUSIONS: We show a massive and selective serum CXCL10 response in R. conorii-infected patients, likely reflecting release from infected endothelial cells characterized by infiltrating T cells and monocytes. The CXCL10 response could contribute to T-cell infiltration within the infected organ, but the pathologic consequences of CXCL10 in clinical R. conorii infection remain to be defined.

Severe Israeli spotted fever with multiorgan failure in a child.

An increased risk of severe and fatal Israeli spotted fever (ISF) has been observed in adults, mostly associated with ISF strain. Here, we report a case of severe ISF with multiorgan failure in a Portuguese child.

[Mediterranean Spotted Fever: Retrospective Review of Hospitalized Cases and Predictive Factors of Severe Disease]. / Febre Escaro-Nodular: Revisão Retrospetiva de Casos Hospitalizados e Fatores Preditores de Doença Severa.

INTRODUCTION: Mediterranean spotted fever, an ancient zoonotic disease raising current issues. This study aims to reevaluate the disease clinical picture and to identify prognostic factors related to severe disease. MATERIAL AND METHODS: We evaluate cases admitted to a central hospital during 12-year period. The risk factors were determined by uni and multivariate analysis, comparing patients admitted in general infirmary versus intermediate/intensive care units or in case of death. RESULTS: We reviewed 71 cases of Mediterranean spotted fever. The mean age was 63.3 ± 16.7 years and 52.1% were male. The eschar was observed in 62.0%. Fever and rash were the most common clinical findings. Forty five percent of patients developed complications and 22.5% were admitted to intermediate and/or intensive care units. Apyrexia, dyspnea, renal failure and elevated lactic dehydrogenase levels at admission were predictors of severe disease development (p < 0.034). Alcoholism was related with prolonged hospitalization (p = 0.020). Absence of fever (p = 0.019) and elevated serum creatinine levels (p = 0.028) were identified as independent factors associated with severe outcome. The mortality rate was 2.8%. DISCUSSION: Although Mediterranean spotted fever is usually assumed to be benign, severe cases are emerging. CONCLUSION: Early identification of the infection and continuous patient surveillance are crucial, particularly in those with greater risk of developing severe or fatal disease.

Introdução: Febre escaro-nodular, uma zoonose antiga a levantar questões actuais. Este estudo pretende reavaliar a imagem clínica da doença e identificar factores prognósticos de doença severa. Material e Métodos: Foram avaliados os casos admitidos num hospital central no período de 12 anos. Os fatores de risco foram determinados por análise uni e multivariada, comparando os indivíduos internados em enfermaria geral versus admitidos em unidades de cuidados intermédios/intensivos ou em caso de morte. Resultados: Foram revistos 71 casos de febre escaro-nodular. A média de idades foi 63,3 ± 16,7 anos e 52,1% eram homens. A escara de inoculação foi identificada em 62,0% dos casos. Febre e exantema foram os achados clínicos mais frequentes. Quarenta e cinco por cento dos doentes desenvolveram complicações e 22,5% foram internados em unidades de cuidados intermédios e/ou intensivos. Apirexia, dispneia, insuficiência renal e níveis de lactato desidrogenase elevados à admissão foram preditores de desenvolvimentode doença severa (p < 0,034). O alcoolismo associou-se a um tempo de internamento prolongado (p = 0,020). Ausênciade febre (p = 0,019) e níveis elevados de creatinina (p = 0,028) constituíram fatores independentes de mau prognóstico. A taxa de mortalidade foi 2,8%.Discussão: Apesar da evolução da febre escaro-nodular ser, geralmente, assumida como benigna, casos graves têm surgido. Quase metade dos doentes hospitalizados desenvolveu complicações. Apirexia e lesão renal predispõem a pior prognóstico. Conclusão: A identificação precoce da infeção e a vigilância contínua dos doentes são determinantes, particularmente naqueles com maior risco de desenvolverem doença severa ou fatal.

Skin lesions and inoculation eschars at the tick bite site in spotted fever group rickettsioses: experience from a patient series in eastern crete, Greece.

The main clinical signs and symptoms caused by a rickettsial infection typically begin 6-10 days after the bite and are accompanied by nonspecific findings such as fever, headache and muscle pain. The diagnosis is mainly based on serological tests, however antibody presentation may be delayed, at least at the early stages of the disease, while seroconversion is usually detected 10-15 days after disease onset. Culture is difficult, requires optimized facilities and often proves negative. Under this scope, the presence of a characteristic inoculation eschar at the bite site may prove a useful clinical tool towards the early suspicion and diagnosis/differential diagnosis of tick-borne rickettsioses, even before the onset of rash and fever or serological confirmation. We describe herein the presence of skin lesions and/or an inoculation eschar at the tick bite site in 17 patients diagnosed, by molecular means, as suffering from spotted fever group rickettsioses. The detection of the pathogen's DNA in biopsy samples proved to be a useful means for early rickettsiae detection and identification. Moreover, the presence of an infiltrated erythema always seemed to precede the appearance of an eschar by 2-5 days and the initiation of fever by 1-10 days; these two signs might also prove useful in the context of the final diagnosis.

Mediterranean spotted fever in children in the Pazardjik region, South Bulgaria.

The objective of the present study was to analyze the clinical pattern of contemporary Mediterranean spotted fever in children and to compare it with the clinical pattern in adults. The research involved 257 children. The more common symptoms in children were severe onset, fever, rash, "tache noire," swelling of lymph nodes and enlarged liver or spleen.

Mediterranean spotted fever in southeastern Romania.

Although cases of Mediterranean spotted fever (MSF) have been reported for decades in southeastern Romania, there are few published data. We retrospectively studied 339 patients, diagnosed with MSF at the National Institute of Infectious Diseases "Prof. Dr. Matei Bals" between 2000 and 2011, in order to raise awareness about MSF in certain regions of Romania. According to the Raoult diagnostic criteria 171 (50.4%) had a score >25 points. Mean age was 52.5 years. One hundred and fifty-five (90.6%) patients were from Bucharest and the surrounding region. Almost all patients presented with fever (99.4%) and rash (98.2%), and 57.9% had evidence of a tick bite. There were no recorded deaths. Serologic diagnosis was made by indirect immunofluorescence assay. Of the 171 patients, serology results for R. conorii were available in 147. One hundred and twenty-three (83.7%) of them had a titer IgG ≥1:160 or a fourfold increase in titer in paired samples. MSF is endemic in southeastern Romania and should be considered in patients with fever and rash even in the absence of recognized tick exposure. Since the disease is prevalent in areas highly frequented by tourists, travel-associated MSF should be suspected in patients with characteristic symptoms returning from the endemic area.

Changes of Bcl-2, Bax and Caspase-3 expression in the dermal microvascular endothelial cells and the epidermal layers of the eschar (tache noire) in patients with Mediterranean spotted fever.

Mediterranean spotted fever (MSF) is widely prevalent in many endemic regions in Bulgaria. The disease is still not quite thoroughly studied as to some aspects of its pathogenesis and especially to issues that concern the crucial signals for apoptosis in the target microvascular endothelial cells. To study the expression of Bcl-2 family proteins and Caspase-3 in the dermal capillary endothelial cells from skin papules and in the eschar (tache noire) epidermal layers of patients with MSF so that we can establish apoptotic processes and the time of their occurrence and deployment. Immunohistochemical reactions for Bcl-2, Bax and Caspase-3 were obtained in slices of punch-biopsies taken from papules of the skin rash and from the eschars of eight patients with MSF. The average intensity of the reactions was compared with that in control punch-biopsy slices from four healthy subjects. MSF was etiologically confirmed in all patients by positive antibody response to a specific antigen, Rickettsia conorii, with indirect immunofluorescent assay performed by the Rickettsial Reference Laboratory. The immune reaction for Bcl-2 was found to be poorly expressed in the capillary endothelial cells of skin papules of patients without any differences from controls. The expression of Bax and Caspase-3 was strongly upregulated in comparison with the controls. The Bcl-2/Bax ratio was significantly decreased. Microvascular endothelial cells of the eschar showed similar changes. While the Bcl-2/Bax ratio decreased in the epidermal layers of the eschar "tache noire", there were no changes in the intensity of the immunoreactivity of Caspase-3 as compared with controls. The upregulation of Bax and Caspase-3 is an indication of ongoing apoptotic processes in the dermal microvascular endothelial cells of MSF patients. The epidermal layers of the eschar showed increased sensitivity to apoptosis, however, executive phase of apoptosis did not occur.

Compartmentalized, functional role of angiogenin during spotted fever group rickettsia-induced endothelial barrier dysfunction: evidence of possible mediation by host tRNA-derived small noncoding RNAs.

BACKGROUND: Microvascular endothelial barrier dysfunction is the central enigma in spotted fever group (SFG) rickettsioses. Angiogenin (ANG) is one of the earliest identified angiogenic factors, of which some are relevant to the phosphorylation of VE-cadherins that serve as endothelial adherens proteins. Although exogenous ANG is known to translocate into the nucleus of growing endothelial cells (ECs) where it plays a functional role, nuclear ANG is not detected in quiescent ECs. Besides its nuclear role, ANG is thought to play a cytoplasmic role, owing to its RNase activity that cleaves tRNA to produce small RNAs. Recently, such tRNA-derived RNA fragments (tRFs) have been shown to be induced under stress conditions. All these observations raise an intriguing hypothesis about a novel cytoplasmic role of ANG, which is induced upon infection with Rickettsia and generates tRFs that may play roles in SFG rickettsioses. METHODS: C3H/HeN mice were infected intravenously with a sublethal dose of R. conorii. At days 1, 3, and 5 post infection (p.i.), liver, lung and brain were collected for immunofluorescence (IF) studies of R. conorii and angiogenin (ANG). Human umbilical vein endothelial cells (HUVECs) were infected with R. conorii for 24, 48, and 72 hrs before incubation with 1µg/ml recombinant human ANG (rANG) in normal medium for 2 hrs. HUVEC samples were subjected to IF, exogenous ANG translocation, endothelial permeability, and immunoprecipitation phosphorylation assays. To identify small non-coding RNAs (sncRNAs) upon rickettsial infection, RNAs from pulverized mouse lung tissues and HUVECs were subjected to library preparation and deep sequencing analysis using an Illumina 2000 instrument. Identified sncRNAs were confirmed by Northern hybridization, and their target mRNAs were predicted in silico using BLAST and RNA hybrid programs. RESULTS: In the present study, we have demonstrated endothelial up-regulation of ANG, co-localized with SFG rickettsial infection in vivo. We also have provided direct evidence that rickettsial infection sensitizes human ECs to the translocation of exogenous ANG in a compartmentalized pattern at different times post-infection. Typically, exogenous ANG translocates into the nucleus at 24 hrs and to the cytoplasm at 72 hrs post-infection. The ANG cytoplasmic translocation enhances phosphorylation and destabilization of VE-cadherin and attenuates endothelial barrier function. Of note, deep sequencing analysis detected tRFs, mostly derived from the 5'-halves of host tRNAs, that are induced by ANG. Northern hybridization validates the two most abundantly cloned tRFs derived from tRNA-ValGTG and tRNA-GlyGCC, in both mouse tissues and human cells. Bioinformatics analysis predicted that these tRFs may interact with transcripts associated with the endothelial barrier, the host cell inflammatory response, and autophagy. CONCLUSIONS: Our data provide new insight into the role of compartmentalized ANG during SFG rickettsioses, and highlight its possible mediation through tRFs.

Gangrene in cases of spotted fever: a report of three cases.

Gangrene is an uncommon complication in cases of rickettsial spotted fever. We report three cases of spotted fever from south India, presumably caused by Rickettsia conorii subspecies indica. Along with gangrene, these cases had severe manifestations of sepsis and multiorgan dysfunction syndrome (MODS) like acute kidney injury, liver dysfunction, delirium and seizure. One patient died while the other two recovered well. This case series is being reported to highlight the occurrence of gangrene in spotted fever rickettsiosis and the importance of appropriate management at the earliest.

Mediterranean spotted fever in the Trakya region of Turkey.

Mediterranean spotted fever (MSF) is caused by a tick-borne pathogen, Rickettsia conorii subsp. conorii, belonging to the spotted fever group (SFG) rickettsiae. The aim of the present study was to evaluate the cases with confirmed diagnosis of MSF from 2003 to 2009 in the Trakya region of Turkey. Patients with high fever, maculopapular rash (involving the palms or soles) and/or a black inoculation eschar at the site of the tick bite (tache noire) were included in the study. Before doxycycline treatment, skin biopsy specimens, preferably from the eschar or from the maculopapular rash, were obtained for DNA extraction. Immunofluorescence assay (IFA) was performed to detect IgM and IgG antibodies against R. conorii in acute and convalescent sera. Afterwards, a standard PCR reaction using primers suitable for hybridisation within the conserved region of genes coding for outer membrane protein A (ompA) and citrate synthase (gltA) and DNA sequencing were performed. There were 128 patients with confirmed MSF diagnosis. Using IFA, seroconversion or a fourfold or greater rise in titre was observed in 97 (77%) patients, whereas a single high titre was demonstrated in 16 (12.7%) patients. According to PCR analysis, 77 (72.6%) of 106 biopsy samples showed positive results. Of these, 58 (73%) of 79 biopsy specimens were from the eschar and 19 (70%) of 27 specimens were from the maculopapular rash. No significant difference was found between the rate of positive skin biopsies taken from the eschar and the maculopapular rash. DNA sequence analysis was performed to all PCR-positive cases, and R. conorii conorii (type strain: Malish, ATCC VR-613) was detected in each of them. MSF is prevalent, but has been underdiagnosed and underreported so far in Turkey. It is a potentially severe and even fatal disease resembling viral haemorrhagic fevers that has to be included in the differential diagnosis of febrile illness associated with thrombocytopenia, even in the absence of an eschar or a tick bite. While IFA allows for retrospective diagnosis in MSF, advanced molecular techniques provide the rapid detection of rickettsia in all skin samples, including eschar and maculopapular rash.

Regulation of inducible heme oxygenase and cyclooxygenase isozymes in a mouse model of spotted fever group rickettsiosis.

Vascular endothelial cells (ECs) lining the blood vessels are the preferred primary targets of pathogenic Rickettsia species in the host. In response to oxidative stress triggered by infection, ECs launch defense mechanisms such as expression of heme oxygenase-1 (HO-1). Previous evidence from an established animal model of Rocky Mountain spotted fever also suggests selective modulation of anti-oxidant enzyme activities in the target host tissues. In this study, we have examined the expression profiles of HO-1 and COX-2 in different tissues during Rickettsia conorii infection of susceptible C3H/HeN mice. RNA hybridization with murine HO-1 and COX-2-specific complementary DNA probes revealed increased HO-1 expression in the liver and brain of mice infected with three different doses of R. conorii ranging from 2.25×10(3) to 2.25×10(5) pfu, relatively non-remarkable changes in the lungs, and a trend for down-regulation in the spleen. The most prominent HO-1 response was evident in the liver with ∼4-fold increase on day 4 post-infection, followed by a decline on day 7. HO-1 expression in the brain, however, peaked with significantly higher levels on day 7. Following infection with both sub-lethal as well as lethal doses of infection, the transcript encoding COX-2 also displayed a pattern of increased expression in the liver and brain. Although immunohistochemical staining revealed increased abundance of HO-1 protein in the liver of infected mice, adjoining serial sections did not exhibit positive staining for COX-2 in infected tissues. The levels of monocyte chemoattractant protein-1 (MCP-1) and keratinocyte-derived cytokine (KC) were significantly higher in the sera of infected mice and corresponded with the onset and severity of the disease. Treatment of infected animals with anti-oxidants α-lipoic acid and N-acetylcysteine and HO inhibitor stannous protoporphyrin (SnPPIX) showed only selective beneficial effects on HO-1 and COX-2 expression in the liver and spleen and serum levels of KC and MCP-1. R. conorii infection of susceptible mice, therefore, results in selective regulation of the expression of HO-1 and COX-2 in a manner dependent on the target host tissue's cellular environment and the propensity of infection with rickettsiae.

Mediterranean spotted fever and encephalitis: a case report and review of the literature.

Mediterranean spotted fever (MSF) is a disease caused by Rickettsia conorii and transmitted by the brown dog tick Rhipicephalus sanguineus. It is widely distributed through southern Europe, Africa, and the Middle East. It is an emerging or a reemerging disease in some regions. Countries of the Mediterranean basin, such as Portugal, have noticed an increased incidence of MSF over the past 10 years. It was believed that MSF was a benign disease associated with a mortality rate of 1-3% before the antimicrobial drug era. It was called benign summer typhus. Severe forms were described in 1981, and the mortality rate reached 32% in Portugal in 1997. However, neurological manifestations associated with brain lesions are a rare event. We describe the case of a man with fever, maculopapular rash, a black spot, and hemisensory loss including the face on the left side of the body with brain lesions in the imaging studies.