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1.
Ticks Tick Borne Dis ; 6(5): 611-4, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26055233

RESUMO

Tularaemia, caused by Francisella tularensis, is an endemic zoonosis frequently occurring in southwest Germany. Since 2005 there is an increase in the number of reported cases of tularaemia in Germany. We report on two cases of ulceroglandular tularaemia and one case of glandular tularaemia that occurred in the summer of 2012 and 2013 in two counties in the Federal State of Baden-Wuerttemberg. Bacteria were transmitted through tick bites, which to date has only rarely been reported in Germany. Inadequate treatment of the patients and an aggravation of clinical symptoms were caused by a delay between disease onset and the detection of the pathogen. Although contact to or consumption of infected hares are the most often reported transmission routes of tularaemia in Germany, tick-bites should also be taken into account. Health professionals should include Francisella tularensis in the differential diagnosis of patients with fever and/or ulcerative lymphadenopathy following a tick bite.


Assuntos
Doenças Transmitidas por Carrapatos/transmissão , Tularemia/epidemiologia , Adulto , Idoso , Animais , Antibacterianos/uso terapêutico , Feminino , Francisella tularensis/isolamento & purificação , Alemanha/epidemiologia , Humanos , Levofloxacino/uso terapêutico , Masculino , Doenças Transmitidas por Carrapatos/epidemiologia , Fatores de Tempo , Tularemia/diagnóstico , Tularemia/tratamento farmacológico , Tularemia/patologia
2.
Genome Announc ; 1(1)2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23405342

RESUMO

Here, we describe the genome sequence of the Francisella tularensis subsp. holarctica strain F92, belonging to the Franco-Iberian subgroup. This strain represents the first-time isolate of this subgroup in Germany and was obtained from naturally infected marmosets.

3.
Vet Microbiol ; 147(3-4): 420-5, 2011 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-20727685

RESUMO

Different species of non-human primates have been exploited as animal disease models for human hantavirus infections. To study the potential risk of natural hantavirus infection of non-human primates, we investigated serum samples from non-human primates of three species living in outdoor enclosures of the German Primate Center (GPC), Göttingen, located in a hantavirus endemic region of central Germany. For that purpose we used serological assays based on recombinant antigens of the bank vole (Myodes glareolus) transmitted Puumala virus (PUUV) and the common and field vole (Microtus arvalis, Microtus agrestis) associated Tula virus (TULV) which are both broadly geographically distributed in Germany. In 24 out of 251 (9.6%) monkey sera collected in 2006 PUUV- and/or TULV-reactive immunoglobulin G (IgG) antibodies were detected. Investigation of follow-up sera from 13 animals confirmed for two animals a seroconversion due to hantavirus exposure at the GPC. To prove the origin of the infection, wild rodents from the surrounding regions were analyzed by hantavirus-specific reverse transcriptase-PCR analysis. In 6 of the 73 investigated bank voles and 3 of the 19 investigated Microtus spp. PUUV- and TULV-specific nucleic acid sequences, respectively, were detected. In conclusion, our investigations demonstrate for the first time natural infections of non-human primates in outdoor enclosures in Germany. These findings highlight the importance of hantavirus surveillance in those primate housings and corresponding preventive measures against wild rodents, particularly in hantavirus endemic regions.


Assuntos
Animais de Zoológico , Arvicolinae/virologia , Cercopithecinae , Infecções por Hantavirus/veterinária , Doenças dos Macacos/epidemiologia , Doenças dos Macacos/virologia , Doenças dos Roedores/virologia , Animais , Anticorpos Antivirais/sangue , Feminino , Alemanha , Orthohantavírus , Infecções por Hantavirus/diagnóstico , Infecções por Hantavirus/epidemiologia , Infecções por Hantavirus/transmissão , Imunoglobulina G/sangue , Masculino , Dados de Sequência Molecular , Doenças dos Macacos/diagnóstico , Doenças dos Macacos/transmissão , Fatores de Risco , Doenças dos Roedores/epidemiologia
4.
J Biomed Inform ; 42(4): 605-11, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19535009

RESUMO

The analysis of large-scale gene expression profiles is still a demanding and extensive task. Modern machine learning and data mining techniques developed in linear algebra, like Independent Component Analysis (ICA), become increasingly popular as appropriate tools for analyzing microarray data. We applied ICA to analyze kinetic gene expression profiles of human monocyte derived macrophages (MDM) from three different donors infected with Francisella tularensis holartica and compared them to more classical methods like hierarchical clustering. Results were compared using a pathway analysis tool, based on the Gene Ontology and the MeSH database. We could show that both methods lead to time-dependent gene regulatory patterns which fit well to known TNFalpha induced immune responses. In comparison, the nonexclusive attribute of ICA results in a more detailed view and a higher resolution in time dependent behavior of the immune response genes. Additionally, we identified NFkappaB as one of the main regulatory genes during response to F. tularensis infection.


Assuntos
Francisella tularensis/fisiologia , Perfilação da Expressão Gênica/métodos , Macrófagos/fisiologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Análise de Componente Principal , Tularemia/genética , Algoritmos , Células Cultivadas , Análise por Conglomerados , Redes Reguladoras de Genes , Humanos , Macrófagos/metabolismo , Macrófagos/microbiologia , Modelos Genéticos , Tularemia/metabolismo
5.
Epidemiol Infect ; 137(5): 736-43, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-18808726

RESUMO

Tularemia is a rare, notifiable zoonosis in Germany. Since November 2004, several lines of evidence including outbreaks in humans or animals and confirmed infections in indigenous hare and rodent populations have indicated a re-emergence of tularemia in different German federal states. Unfortunately, reliable basic information on the seroprevalence in different geographical regions, permitting the identification of risk factors, does not exist. Combining a sensitive screening assay with a highly specific confirmative immunoblot test, we performed a serological investigation on 2416 sera from a population-based, cross-sectional health survey of the city population of Leutkirch, Baden-Wuerttemberg. A total of 56 sera gave positive results indicating a seroprevalence of 2.32%. Thus, the seroprevalence is tenfold higher than that previously reported in a nationwide study in 2004. Francisella tularensis can cause a wide variety of clinical syndromes including severe, sometimes fatal disease. Missing epidemiological data on its spatial and temporal distribution in an endemic country complicate an appropriate risk assessment necessary for public health authorities to be prepared for an adequate outbreak management. This is of special concern regarding the extraordinary potential of F. tularensis as an agent of bioterrorism. Our investigation performed in a presumed low-risk area demonstrated that tularemia might be seriously underestimated in Germany and probably in other central European countries as well.


Assuntos
Francisella tularensis/isolamento & purificação , Tularemia/epidemiologia , Adolescente , Adulto , Idoso , Animais , Anticorpos Antibacterianos/sangue , Criança , Estudos Transversais , Feminino , Alemanha/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Adulto Jovem
6.
Trans R Soc Trop Med Hyg ; 102 Suppl 1: S40-1, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19121684

RESUMO

A 62-year-old German patient with insulin-dependent diabetes and diverticulitis was hospitalized for abdominal pain of the left lower quadrant. Further examination revealed an abdominal abscess, which was punctured. Presumptively a Pseudomonas species was identified, but further examination revealed Burkholderia pseudomallei as the causative agent. Most probably this infection was acquired in 1996 during a trip to Thailand, where the patient had been hospitalized. After combined chemotherapy and surgical revision of the abscess, the patient's condition improved. Clinicians and microbiologists have to keep in mind that in some tropical infections such as melioidosis relapse may occur after such a long time.


Assuntos
Abscesso Abdominal/diagnóstico , Burkholderia pseudomallei/isolamento & purificação , Melioidose/diagnóstico por imagem , Burkholderia pseudomallei/genética , Diagnóstico Precoce , Alemanha , Humanos , Pessoa de Meia-Idade , Recidiva , Tailândia , Viagem , Ultrassonografia
7.
J Clin Microbiol ; 45(10): 3404-7, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17652472

RESUMO

An in-house immunochromatographic test, Plague BioThreat Alert test strips, ABICAP columns, enzyme-linked immunosorbent assay, flow cytometry, and immunofluorescence microscopy were compared for the detection of the fraction 1 capsular antigen of Yersinia pestis, using spiked buffer and clinical specimens. Hand-held test kits proved to be excellent benchtop tools.


Assuntos
Proteínas de Bactérias/análise , Ensaio de Imunoadsorção Enzimática/métodos , Citometria de Fluxo/métodos , Kit de Reagentes para Diagnóstico , Yersinia pestis/isolamento & purificação , Microscopia de Fluorescência
8.
Vet Pathol ; 44(3): 327-34, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17491074

RESUMO

Tularemia is a highly contagious infectious zoonosis, transmissible by inoculation, ingestion, or inhalation of the infectious agent Francisella tularensis. The disease is perpetuated by infected rodents, blood-sucking arthropods, and by contaminated water. Therefore, nonhuman primates housed outdoors may be at risk for exposure. An epizootic of F. tularensis occurred in an indoor/outdoor-housed group of cynomolgus monkeys (Macaca fascicularis) at the German Primate Center. Tularemia was diagnosed in 18 out of 35 animals within a period of 2 years. Six animals died with unspecific clinical symptoms; 12 animals developed seroconversion and were still alive. Pathologic findings were similar in all monkeys that died and resembled the clinical picture of the human disease, including an ulceroglandular syndrome with local lymphadenopathy, gingivostomatitis, and systemic spread, with manifestations such as subacute necrotizing hepatitis, granulomatous splenitis, and pneumonia. Tularemia was diagnosed by culture, real-time polymerase chain reaction, and ELISA techniques. This is the largest outbreak in nonhuman primates and the first report of tularemia in cynomolgus monkeys. An overview of the recent literature about tularemia in nonhuman primates is given.


Assuntos
Macaca fascicularis , Doenças dos Macacos/diagnóstico , Tularemia/veterinária , Animais , Feminino , Gengivite/patologia , Gengivite/veterinária , Abrigo para Animais , Rim/patologia , Fígado/patologia , Pulmão/patologia , Masculino , Doenças dos Macacos/patologia , Baço/patologia , Língua/patologia , Tularemia/diagnóstico , Tularemia/patologia
9.
Epidemiol Infect ; 135(8): 1256-65, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17306050

RESUMO

Francisella tularensis was identified as the cause of a die-off which occurred among a colony of semi-free-living common marmosets (Callithrix jacchus). During the outbreak 5 out of 62 animals died of tularaemia in a research facility located in the district of Goettingen, Germany. All animals had been born at the facility suggesting an endemic infection. A total of five culture isolates were recovered and characterized as F. tularensis holarctica, biovar I. These cultures represent the first isolates obtained in the Federal Republic of Germany for more than 45 years. The outbreak area shows several geographical and ecological characteristics known to favour long-term presence of F. tularensis. Persistence of the pathogen in the remote region along the former German-German border, continuous re-introduction from eastern European countries after destruction of the 'Iron curtain' or introduction through migrating birds are testable hypotheses which could explain the emergence of tularaemia in this particular region.


Assuntos
Callithrix/microbiologia , Francisella tularensis/isolamento & purificação , Tularemia/epidemiologia , Tularemia/veterinária , Animais , Técnicas de Tipagem Bacteriana , Feminino , Geografia , Alemanha/epidemiologia , Fígado/microbiologia , Masculino , Reação em Cadeia da Polimerase , Baço/microbiologia , Tularemia/microbiologia
10.
J Vet Med B Infect Dis Vet Public Health ; 52(10): 444-55, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16364020

RESUMO

Brucellosis and tularemia are classical zoonotic diseases transmitted from an animal reservoir to humans. Both, wildlife and domestic animals, contribute to the spreading of these zoonoses. The surveillance of the animal health status is strictly regulated for domestic animals, whereas systematic disease monitoring in wildlife does not exist. The aim of the present study was to provide data on the prevalence of anti-Brucella, anti-Francisella and anti-Yersinia antibodies in wild boars from North-Eastern Germany to assess public health risks. A total of 763 sera of wild boars from Mecklenburg-Western Pomerania hunted in 1995/1996 were tested using a commercially available Brucella suis ELISA, an in-house lipopolysaccharide (LPS)-based Francisella ELISA, and commercially available Western blot kits for the detection of anti-Francisella and anti-Yersinia antibodies. The Yersinia enterocolitica O:9 LPS is able to induce serological cross-reactions indistinguishable from brucellosis due to a similar immunodominant epitope in the Brucella O-polysaccharide. The Yersinia Western blot assay was, therefore, based on five recombinant Yersinia outer proteins which have been proved to be specific for the serodiagnosis of yersiniosis. Anti-Brucella, anti-Francisella and anti-Yersinia antibodies were detected in 22.0%, 3.1%, and 62.6% of the wild boars, respectively. The high seroprevalence of tularemia and brucellosis in wild boars indicates that natural foci of these zoonoses are present in wildlife in Germany. However, the impact of transmission of zoonotic pathogens from wildlife to livestock is unknown. Only careful and systematic monitoring will help to prevent the (re)emergence of these zoonotic diseases in domestic animals and consequently human infection.


Assuntos
Anticorpos Antibacterianos/sangue , Brucelose/veterinária , Sus scrofa , Doenças dos Suínos/epidemiologia , Tularemia/veterinária , Yersiniose/veterinária , Animais , Brucella/imunologia , Brucelose/sangue , Brucelose/epidemiologia , Brucelose/transmissão , Reservatórios de Doenças/veterinária , Feminino , Francisella tularensis/imunologia , Alemanha/epidemiologia , Masculino , Saúde Pública , Estudos Soroepidemiológicos , Doenças dos Suínos/sangue , Doenças dos Suínos/transmissão , Tularemia/sangue , Tularemia/epidemiologia , Tularemia/transmissão , Yersinia/imunologia , Yersiniose/sangue , Yersiniose/epidemiologia , Yersiniose/transmissão , Zoonoses
11.
Artigo em Inglês | MEDLINE | ID: mdl-16219088

RESUMO

Tularaemia is a severe bacterial zoonosis caused by the highly infectious agent Francisella tularensis. It is endemic in countries of the northern hemisphere ranging from North America to Europe, Asia and Japan. Very recently, Francisella-like strains causing disease in humans were described from tropical northern Australia. In the last decade, efforts have been made to develop sensitive and specific immunological and molecular techniques for the laboratory diagnosis of tularaemia and also for the definite identification of members of the species F. tularensis and its four subspecies. Screening for the keyword 'Francisella' a Medline search over the last decade was performed and articles describing diagnostic methods for tularaemia and its causative agent were selected. Besides classical microbiological techniques (cultivation, biochemical profiling, susceptibility testing) several new immunological and molecular approaches to identify F. tularensis have been introduced employing highly specific antibodies and various polymerase chain reaction (PCR)-based methods. Whereas direct antigen detection by enzyme-linked immunosorbent assay (ELISA) or immunofluorescence might allow early presumptive diagnosis of tularaemia, these methods--like all PCR techniques--still await further evaluation. Therefore, diagnosis of tularaemia still relies mainly on the demonstration of specific antibodies in the host. ELISA and immunoblot methods started to replace the standard tube or micro-agglutination assays. However, the diagnostic value of antibody detection in the very early clinical phase of tularaemia is limited. Francisella tularensis is regarded as a 'highest priority' biological agent (category 'A' according to the CDC, Atlanta, GA, USA), thus rapid and reliable diagnosis of tularaemia is required not only for a timely onset of therapy, the handling of outbreak investigations but also for the surveillance of endemic foci. Only very recently, evaluated test kits for serological diagnosis of human tularaemia became available, while the introduction of standardized molecular techniques for detection and typing is still missing.


Assuntos
Francisella tularensis/isolamento & purificação , Tularemia/veterinária , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Francisella tularensis/genética , Francisella tularensis/imunologia , Humanos , Reação em Cadeia da Polimerase/veterinária , Tularemia/diagnóstico , Zoonoses
12.
Cytometry A ; 53(2): 88-96, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12766970

RESUMO

BACKGROUND: Plague is a severe, highly communicable bacterial disease caused by Yersinia pestis. It is still endemic in more than 20 countries worldwide. Although known as a devastating disease for centuries, laboratory confirmation of clinical suspected cases is still problematic. No standardized and internationally approved test system is commercially available. The aim of this study was the introduction and evaluation of a combination of immunomagnetic separation and flow cytometry for the serodiagnosis of human plague. METHODS: Paramagnetic polystyrene beads were coated with purified F1 capsular antigen (F1 CA) and reacted with sera from plague patients, from 26 laboratory personnel vaccinated against plague and from 102 healthy blood donors (HBD). After incubation with fluorescein isothiocyanate-conjugated anti-human rabbit IgG, particle-associated fluorescence was detected by flow cytometry. RESULTS: Anti-F1 CA antibodies could be demonstrated in all patients with bacteriologically confirmed plague and in 22 sera (84.6%) from vaccinees. Only one serum in the HBD group showed a weakly positive reaction. The total assay time was less than 2 h. CONCLUSIONS: Compared with a recently published combination of an anti-F1 CA enzyme-linked immunosorbent assay (ELISA) and immunoblot, the new assay showed the same sensitivity as the ELISA and almost the same specificity (99.0 versus 100%) as the immunoblot. Allowing a rapid, reliable, and quantitative analysis, immunomagnetic separation combined with flow cytometry might replace both conventional immunoassays.


Assuntos
Anticorpos/análise , Citometria de Fluxo/métodos , Separação Imunomagnética/métodos , Peste/sangue , Peste/diagnóstico , Testes Sorológicos/métodos , Anticorpos/sangue , Anticorpos/imunologia , Proteínas de Bactérias/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Immunoblotting , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Fatores de Tempo , Yersinia pestis/imunologia
13.
FEMS Immunol Med Microbiol ; 30(1): 53-63, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11172992

RESUMO

Burkholderia pseudomallei is the etiological agent of melioidosis, a potentially fatal disease occurring in man and animals. The aim of this study was to investigate the pathophysiological course of experimental melioidosis, and to identify the target organs, in an animal model. For this purpose SWISS mice were infected intraperitoneally with the virulent strain B. pseudomallei 6068. The bacterial load of various organs was quantified daily by bacteriological analysis and by an enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody specific to B. pseudomallei exopolysaccharide (EPS). Electron microscopic investigation of the spleen was performed to locate the bacteria at the cellular level. In this model of acute melioidosis, B. pseudomallei had a marked organ tropism for liver and spleen, and showed evidence of in vivo growth with a bacterial burden of 1.6x10(9) colony forming units (CFU) per gram of spleen 5 days after infection with 200 CFU. The highest bacterial loads were detected in the spleen at all time points, in a range from 2x10(6) to 2x10(9) CFU g(-1). They were still 50-80 times greater than the load of the liver at the time of peak burden. Other investigated organs such as lungs, kidneys, and bone marrow were 10(2)-10(4)-fold less infected than the spleen, with loads ranging from 3x10(2) to 3x10(6) CFU g(-1). The heart and the brain were sites of a delayed infection, with counts in a range from 10(3) to 10(7) times lower than bacterial counts in the spleen. The EPS-specific ELISA proved to be highly sensitive, particularly at the level of those tissues in which colony counting on agar revealed low contamination. In the blood, EPS was detected at concentrations corresponding to bacterial loads ranging from 8x10(3) to 6x10(4) CFU ml(-1). Electron microscopic examination of the spleen revealed figures of phagocytosis, and the presence of large numbers of intact bacteria, which occurred either as single cells or densely packed into vacuoles. Sparse figures suggesting bacterial replication were also observed. In addition, some bacteria could be seen in vacuoles that seemed to have lost their membrane. These observations provide a basis for further investigations on the pathogenesis of the disease.


Assuntos
Burkholderia pseudomallei , Melioidose/fisiopatologia , Animais , Burkholderia pseudomallei/crescimento & desenvolvimento , Burkholderia pseudomallei/isolamento & purificação , Burkholderia pseudomallei/patogenicidade , Contagem de Colônia Microbiana , Meios de Cultura , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Melioidose/microbiologia , Melioidose/patologia , Camundongos , Baço/ultraestrutura
14.
Hybridoma ; 17(2): 143-50, 1998 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9627054

RESUMO

Monoclonal antibodies (MAbs) to Burkholderia cepacia were produced from mice immunized with inactivated whole-cell antigen. For screening of resulting MAbs an enzyme-linked immunosorbent assay (ELISA) was used. A stable hybridoma cell line (BC-2) producing specific antibodies to a 64 kDa somatic antigen from B. cepacia was established. In ELISA and immunoblotting analysis the MAb BC-2 recognized all tested strains of B. cepacia whereas no cross-reaction with 32 Pseudomonas aeruginosa strains was found. From a wide range of other bacteria only strains of the species Burkholderia mallei, Burkholderia pseudomallei, and Burkholderia gladioli showed cross-reactions. The MAb BC-2 will be used to develop a diagnostic assay for the identification of B. cepacia and B. gladioli, important agents of nosocomial infections in immunocompromised patients suffering especially from cystic fibrosis (CF).


Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Antígenos de Bactérias/imunologia , Burkholderia cepacia/imunologia , Animais , Anticorpos Monoclonais/biossíntese , Especificidade de Anticorpos , Ensaio de Imunoadsorção Enzimática , Hibridomas/imunologia , Immunoblotting , Camundongos , Especificidade da Espécie
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