RESUMEN
AIMS: To discuss together the novel cases of Brucella infections in frogs with the results of published reports to extend our current knowledge on 'atypical' brucellae isolated from amphibians and to discuss the challenges we face on this extraordinary emerging group of pathogens. METHODS AND RESULTS: Since our first description, an additional 14 isolates from four different frog species were collected. Novel isolates and a subset of Brucella isolates previously cultured from African bullfrogs were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), Fourier transform-infrared (FT-IR) spectroscopy and broth microdilution susceptibility testing. MALDI-TOF MS worked very efficiently for an accurate bacterial identification to the genus level. Within the cluster analysis, 'atypical' brucellae grouped distant from Brucella melitensis and were even more separated by FT-IR spectroscopy with respect to their geographical origin. Minimum inhibitory concentrations of 14 antimicrobial substances are provided as baseline data on antimicrobial susceptibility. CONCLUSIONS: The case history of Brucella infections in amphibians reveals a variety of pathologies ranging from localized manifestations to systemic infections. Some isolates seem to be capable of causing high mortality in zoological exhibitions putting higher demands on the management of endangered frog species. There is considerable risk in overlooking and misidentifying 'atypical' Brucella in routine diagnostics. SIGNIFICANCE AND IMPACT OF THE STUDY: Brucella have only recently been described in cold-blooded vertebrates. Their presence in frog species native to Africa, America and Australia indicates a more common occurrence in amphibians than previously thought. This study provides an extensive overview of amphibian brucellae by highlighting the main features of their clinical significance, diagnosis and zoonotic potential.
Asunto(s)
Brucella/clasificación , Brucella/aislamiento & purificación , Brucelosis/veterinaria , Anfibios , Animales , Australia , Brucella/efectos de los fármacos , Brucella/fisiología , Brucelosis/epidemiología , Brucelosis/microbiología , Brucelosis/patología , Análisis por Conglomerados , Humanos , Pruebas de Sensibilidad Microbiana , Filogenia , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Espectroscopía Infrarroja por Transformada de Fourier , ZoonosisRESUMEN
Brucellosis is a widespread zoonotic disease introduced from animal reservoirs to humans. In Germany, bovine and ovine/caprine brucellosis were eradicated more than a decade ago and mandatory measures in livestock have been implemented to keep the officially brucellosis-free status. In contrast, surveillance of wildlife is still challenging, and reliable data on the prevalence of brucellae in small mammal populations do not exist. To assess the epidemiology of Brucella spp. in rodents and shrews, a molecular survey was carried out. A total of 537 rodents and shrews were trapped in four federal states located throughout Germany and investigated for the presence of Brucella. Using a two-step molecular assay based on the detection of the Brucella-specific bcsp31 and IS711 sequences in tissue samples, 14.2% (n = 76) of the tested animals were positive. These originated mainly from western and south-western Germany, where preliminary analyses indicate population density-dependent Brucella prevalence in voles (Myodes glareolus) and mice (Apodemus spp.). recA typing revealed a close relationship to a potentially novel Brucella species recently isolated from red foxes (Vulpes vulpes) in Austria. The molecular detection of brucellae in various rodent taxa and for the first time in shrew species shows that these animals may be naturally infected or at least have a history of exposure to Brucella spp.
Asunto(s)
Brucella/aislamiento & purificación , Brucelosis/epidemiología , Reservorios de Enfermedades , Roedores/microbiología , Musarañas/microbiología , Animales , Brucelosis/veterinaria , Alemania/epidemiologíaRESUMEN
The genus Brucella (Mayer and Shaw, 1920) currently consists often species with validly published names. Within most species further differentiation into biovars exists. Genetically, all Brucella species are highly related to each other, exhibiting sequence similarity values of 98% to 100% in aligned regions (core genome). The population structure is clonal. Despite this close genetic relatedness, the various species can be clearly distinguished from each other by application of high-resolution molecular typing tools, in addition to assessment of phenotype and host preference. Accurate species delineation can be achieved by conventional multiplex polymerase chain reaction (PCR), single nucleotide polymorphism (SNP) analysis and multilocus sequence typing (MLST) or multilocus sequence analysis (MLSA). The last is also suitable for phylogenetic reconstructions, owing to the highly clonal evolution of the different species. Highly discriminatory multilocus variable number of tandem repeats (VNTR) analysis (MLVA) allows both species delineation and differentiation of individual isolates and thus represents a perfect first-line toolfor molecular epidemiological studies within outbreak investigations. More recently,whole genome sequencing (WGS)and the resulting global genome-wide SNP analysis have become available. These novel approaches should help in further understanding the evolution, host specificity and pathogenicity of the genus Brucella.
Asunto(s)
Brucella/clasificación , Brucella/genética , Bases de Datos Factuales , Regulación Bacteriana de la Expresión Génica , Genoma Bacteriano/genética , Tipificación de Secuencias Multilocus , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Nucleótido Simple , Especificidad de la Especie , Secuencias Repetidas en TándemRESUMEN
A gram-negative, rod-shaped, non-spore-forming bacterium, isolated from placental tissue of a cow, was investigated for its taxonomic position. On the basis of 16S rRNA gene sequence similarities, strain UK34/07-5(T) was shown to belong to the class Alphaproteobacteria, closely related to the type strain of Camelimonas lactis (96.0â% sequence similarity). The polyamine pattern showed the major compound spermidine and moderate amounts of putrescine. The major quinone was ubiquinone Q-10. The polar lipid profile was composed of the major compounds phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol and phosphatidylmonomethylethanolamine and moderate amounts of diphosphatidylglycerol, three unidentified aminolipids and an unidentified phospholipid. The profile of major fatty acids, consisting of C(19â:â0) cyclo ω8c and C(18â:â1)ω7c, with C(18â:â0) 3-OH as the hydroxylated fatty acid, was very similar to that of C. lactis M 2040(T). The results of DNA-DNA hybridization and physiological and biochemical tests allowed both genotypic and phenotypic differentiation of the isolate from C. lactis. The relatively low 16S rRNA gene sequence similarity of 96.0â% to C. lactis M 2040(T) and marked differences in the polar lipid profiles as well as the results of physiological tests and the DNA-DNA hybridization data support the creation of a novel species, for which the name Camelimonas abortus sp. nov. is proposed, with the type strain UK34/07-5(T) (â=âCIP 110303(T) â=âCCUG 61094(T) â=âDSM 24741(T) â=âCCM 7941(T)).
Asunto(s)
Beijerinckiaceae/clasificación , Beijerinckiaceae/aislamiento & purificación , Enfermedades de los Bovinos/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Placenta/microbiología , Animales , Técnicas de Tipificación Bacteriana , Beijerinckiaceae/química , Beijerinckiaceae/genética , Bovinos , Análisis por Conglomerados , Citosol/química , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Femenino , Infecciones por Bacterias Gramnegativas/microbiología , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Fosfolípidos/análisis , Filogenia , Poliaminas/análisis , Embarazo , Quinonas/análisis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Following the recent discovery of new Brucella strains from different animal species and from the environment, ten Brucella species are nowadays included in the genus Brucella. Although the intracellular trafficking of Brucella is well described, the strategies developed by Brucella to survive and multiply in phagocytic and non-phagocytic cells, particularly to access nutriments during its intracellular journey, are still largely unknown. Metabolism and virulence of Brucella are now considered to be two sides of the same coin. Mechanisms presiding to the colonization of the pregnant uterus in different animal species are not known. Vaccination is the cornerstone of control programs in livestock and although the S19, RB51 (both in cattle) and Rev 1 (in sheep and goats) vaccines have been successfully used worldwide, they have drawbacks and thus the ideal brucellosis vaccine is still very much awaited. There is no vaccine available for pigs and wildlife. Animal brucellosis control strategies differ in the developed and the developing world. Most emphasis is put on eradication and on risk analysis to avoid the re-introduction of Brucella in the developed world. Information related to the prevalence of brucellosis is still scarce in the developing world and control programs are rarely implemented. Since there is no vaccine available for humans, prevention of human brucellosis relies on its control in the animal reservoir. Brucella is also considered to be an agent to be used in bio- and agroterrorism attacks. At the animal/ecosystem/human interface it is critical to reduce opportunities for Brucella to jump host species as already seen in livestock, wildlife and humans. This task is a challenge for the future in terms of veterinary public health, as for wildlife and ecosystem managers and will need a "One Health" approach to be successful.
Asunto(s)
Brucella/aislamiento & purificación , Brucelosis/veterinaria , Zoonosis/microbiología , Animales , Brucelosis/epidemiología , Brucelosis/microbiología , Femenino , Humanos , Embarazo , Zoonosis/epidemiologíaRESUMEN
Three strains of Gram-negative, rod-shaped, non-spore-forming bacteria (M 2040(T), M 1973 and M 1878-SK2), isolated from milk of camels at a camel-milk production farm in the United Arab Emirates, were investigated for their taxonomic allocation. On the basis of 16S rRNA gene sequence similarities, all three strains were shown to belong to the Alphaproteobacteria and were most closely related to Chelatococcus asaccharovorans and Chelatococcus daeguensis (95.1 and 95.2â% sequence similarity to the respective type strains). meso-Diaminopimelic acid was detected as the characteristic peptidoglycan diamino acid. The predominant compound in the polyamine pattern was spermidine, and sym-homospermidine was not detectable. The quinone system was ubiquinone Q-10. The polar lipid profile included the major compounds phosphatidylcholine and diphosphatidylglycerol and moderate amounts of phosphatidylethanolamine, phosphatidylglycerol, an unidentified glycolipid and two unidentified aminolipids. Minor lipids were also detected. The major fatty acid profile, consisting of C19â:0 cyclo ω8c and C18:1 ω7c, with C18â:03-OH as the major hydroxylated fatty acid, was similar to those of the genus Chelatococcus. The results of DNA-DNA hybridization experiments and physiological and biochemical tests allowed both genotypic and phenotypic differentiation of the isolates from described Chelatococcus species. Isolates M 2040(T), M 1973 and M 1878-SK2 were closely related on the basis of DNA-DNA reassociation and therefore represent a single novel species. In summary, low 16S rRNA gene sequence similarities of 95â% with Chelatococcus asaccharovorans and marked differences in polar lipid profiles as well as in polyamine patterns support the description of a novel genus and species to accommodate these strains, for which the name Camelimonas lactis gen. nov., sp. nov. is proposed. The type strain of Camelimonas lactis is M 2040(T) (=CCUG 58638(T) =CCM 7696(T)).
Asunto(s)
Beijerinckiaceae/clasificación , Beijerinckiaceae/aislamiento & purificación , Camelus/microbiología , Leche/microbiología , Animales , Técnicas de Tipificación Bacteriana , Beijerinckiaceae/química , Beijerinckiaceae/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácido Diaminopimélico/análisis , Ácidos Grasos/análisis , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Peptidoglicano/química , Fosfolípidos/análisis , Filogenia , Poliaminas/análisis , Quinonas/análisis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Emiratos Árabes UnidosRESUMEN
A systemic disease occurred in a wild population of the common vole Microtus arvalis in South Moravia (Czech Republic) during the years 1999-2003. Acute infections were characterized by edema of extremities, occasionally with colliquating abscesses, arthritis, lymphadenitis, perforations of the skin resulting from colliquated abscesses, orchitis, and peritoneal granulomas. From the clinical samples, small Gram-negative coccobacilli were isolated and identified as Ochrobactrum intermedium by API 20NE and colistin sensitivity profiles. However, subsequent rrs (16S rRNA) and recA (recombinase A) gene sequencing analysis of two isolates (CCM 4915=CAPM 6434; CCM 4916=CAPM 6435) identified them as Brucella sp. with sequence identities of 100% to other Brucella spp. Analysis of the omp2a/b genes confirmed the two isolates as Brucella. In AMOS polymerase chain reaction (PCR), a 2000-bp fragment was generated that was not seen in other brucellae. Experimental infection of outbred ICR mice with these isolates resulted in a mortality rate of 50%. Based on the results of the molecular investigations and the mortality observed in experimentally infected mice we conclude that the epizootic was caused by Brucella sp. and not by Ochrobactrum intermedium. The study demonstrates the limitations of commercial biochemical test systems in accurately differentiating among Ochrobactrum and Brucella.
Asunto(s)
Arvicolinae/microbiología , Brucella/aislamiento & purificación , Brucella/fisiología , Brucelosis/veterinaria , Enfermedades de los Roedores/microbiología , Animales , Sangre/microbiología , Brucella/clasificación , Brucella/genética , Brucelosis/microbiología , Brucelosis/patología , República Checa/epidemiología , Femenino , Ganglios Linfáticos/microbiología , Masculino , Ratones , Ratones Endogámicos ICR , Datos de Secuencia Molecular , Ochrobactrum/clasificación , Ochrobactrum/aislamiento & purificación , Enfermedades de los Roedores/patologíaRESUMEN
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.
Asunto(s)
Proteínas Bacterianas/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Citometría de Flujo/métodos , Juego de Reactivos para Diagnóstico , Yersinia pestis/aislamiento & purificación , Microscopía FluorescenteRESUMEN
A PCR assay targeting the metalloprotease gene (mprA) of Burkholderia pseudomallei was developed for the specific detection of this organism in pure cultures and clinical samples. All other closely related organisms including B. mallei the causative agent of glanders, and B. thailandensis tested negative. Burkholderia pseudomallei DNA was successfully amplified from paraffin-embedded lung tissue of a camel with a generalized B. pseudomallei infection. The developed PCR assay can be used as a simple tool for the specific and sensitive detection of B. pseudomallei.
Asunto(s)
Burkholderia pseudomallei/aislamiento & purificación , Metaloproteasas/genética , Reacción en Cadena de la Polimerasa/veterinaria , Animales , Secuencia de Bases , Cartilla de ADN , ADN Bacteriano/química , ADN Bacteriano/genética , Amplificación de Genes , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Especificidad de la EspecieRESUMEN
Since 1990 the number of glanders outbreaks in race, military and pleasure horses in Asia and South America is steadily increasing. Glanders, which is eradicated in Western Europe, Australia and Northern America, is currently considered a re-emerging disease. Consequently, the disease may be introduced into glanders-free regions by subclinical carriers at any time. The causative agent of glanders, Burkholderia (B.) mallei, is highly contagious and leads to chronic disease in horses whereas in donkeys and mules the disease is acute and often fatal. Occurrence of the disease leads to international trading restrictions and infected animals immediately have to be culled and safely disposed off. In humans B. mallei infection results in a severe clinical course, and is fatal without appropriate therapy. Its pathogenicity makes B. mallei a potential biological agent that may be used in bioterroristic attacks. Due to the eradication of glanders in the second half of the last century, veterinarians in western European countries are no longer familiar with its clinical presentation in solipeds. Having these facts in mind, this review describes the epidemiology, clinical signs, pathology and the current eradication strategy of this interesting zoonosis. Pictures of imported endurance horses infected with glanders taken during an eradication campaign in Dubai, United Arab Emirates, in 2004 illustrate most typical clinical findings.
Asunto(s)
Brotes de Enfermedades/veterinaria , Equidae , Muermo/epidemiología , Muermo/prevención & control , Zoonosis , Animales , Bioterrorismo , Burkholderia mallei/patogenicidad , Diagnóstico Diferencial , Brotes de Enfermedades/prevención & control , Muermo/transmisión , Caballos , Humanos , Cooperación InternacionalRESUMEN
Yersiniosis is caused by Y. enterocolitica and Y. pseudotuberculosis mostly presenting as intestinal infection. The infection is usually acquired from contaminated food. The aim of this study was to determine the seroprevalence of anti-Yersinia antibodies in Austrians. Sera of 750 healthy Austrians from all nine states were tested for anti-Yersinia IgG antibodies using the recomBlot Yersinia Westernblot kit. Overall seroprevalence was 29.7%. Seroprevalence increased significantly with age from 24.7% in the group of the 19 to 24 year olds to 38.5% in the group of persons older than 44 years. The seroprevalence of anti-Yersinia antibodies varied within the states between 18% and 43.5%. The high seroprevalence of anti-Yersinia antibodies in contrast to only approximately 100 reported yersiniosis cases per year points to the fact that the majority of infections is either subclinical or mild.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Yersiniosis/epidemiología , Yersinia enterocolitica/inmunología , Yersinia pseudotuberculosis/inmunología , Adulto , Distribución por Edad , Austria/epidemiología , Proteínas de la Membrana Bacteriana Externa/inmunología , Western Blotting , Femenino , Humanos , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Estudios Seroepidemiológicos , Yersiniosis/diagnóstico , Yersiniosis/inmunologíaRESUMEN
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.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Brucelosis/veterinaria , Sus scrofa , Enfermedades de los Porcinos/epidemiología , Tularemia/veterinaria , Yersiniosis/veterinaria , Animales , Brucella/inmunología , Brucelosis/sangre , Brucelosis/epidemiología , Brucelosis/transmisión , Reservorios de Enfermedades/veterinaria , Femenino , Francisella tularensis/inmunología , Alemania/epidemiología , Masculino , Salud Pública , Estudios Seroepidemiológicos , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/transmisión , Tularemia/sangre , Tularemia/epidemiología , Tularemia/transmisión , Yersinia/inmunología , Yersiniosis/sangre , Yersiniosis/epidemiología , Yersiniosis/transmisión , ZoonosisRESUMEN
Burkholderia mallei causes glanders or farcy in solipeds, a disease that must be reported to the OIE (Office International des Epizooties, Paris, France). The number of reported outbreaks has increased steadily during the last decade. Serodiagnosis is hampered by the considerable number of false-positives and -negatives of the internationally prescribed tests. The major problem leading to low sensitivity and specificity of complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e. crude preparations of whole cells. Future perspectives for the development and evaluation of serological test kits using well-characterized single antigens are discussed in the light of recent molecular research on B. mallei and the closely related saprozoonotic agent B. pseudomallei.
Asunto(s)
Infecciones por Burkholderia/veterinaria , Burkholderia mallei/inmunología , Enfermedades de los Caballos/diagnóstico , Animales , Infecciones por Burkholderia/diagnóstico , Burkholderia mallei/aislamiento & purificación , Pruebas de Fijación del Complemento/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Enfermedades de los Caballos/sangre , Caballos , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad , Pruebas Serológicas/métodos , Pruebas Serológicas/veterinariaRESUMEN
Human brucellosis has become a rare disease in Germany since the eradication of bovine and ovine/caprine brucellosis in this country. Therefore, most physicians are unfamiliar with the illnesses clinical presentation, diagnostic tools, and therapeutic strategies. This retrospective study was carried out to evaluate the epidemiological, clinical, and laboratory features of human brucellosis in Germany in the years 2002 and 2003. Thirty-one bacterial isolates from 30 patients sent to the German national reference laboratory were characterized using the genus-specific bcsp31 real-time PCR, the species-specific AMOS-PCR, and standard microbiological methods for the detection and identification of Brucella spp. The medical records of all patients with bacteriologically confirmed brucellosis were evaluated. All 31 isolates proved to be Brucella (30 Brucella melitensis and 1 Brucella suis). Most of the brucellosis patients were infected in endemic countries while visiting friends and relatives during their summer holidays. One case of laboratory-acquired infection was identified. Brucellosis was transmitted mainly by the consumption of contaminated unpasteurized milk or cheese from goats and sheep. The patients presented primarily with flu-like symptoms, i.e. fever, chills, sweating, headaches, arthralgia, and myalgia. In most cases, however, symptoms and signs of focal complications, e.g. spondylitis, endocarditis, and meningoencephalitis, predominated. The rate of complications was much higher than that in endemic countries, presumably as a result of diagnostic delay due to a low index of suspicion. In summary, physicians in nonendemic countries such as Germany must be aware of brucellosis being a possible cause of fever of unknown origin in immigrants and tourists travelling from endemic countries.
Asunto(s)
Brucelosis/epidemiología , Adolescente , Adulto , Factores de Edad , Anciano , Brucelosis/diagnóstico , Brucelosis/fisiopatología , Niño , Enfermedades Transmisibles Emergentes , Femenino , Alemania/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
INTRODUCTION: In spite of new surgical techniques and recently developed antibiotics, there is no satisfactory solution for the treatment of chronic posttraumatic osteomyelitis. The introduction of local antibiotic treatment with gentamicin-PMMA beads according to Klemm has provided new stimuli for the treatment of chronic osteomyelitis. With the development of collagen as an absorbable carrier substance, the disadvantages of the rigid carrier system became evident. Due to the varying surgical techniques and different forms of adjuvant therapy, it is difficult to assess therapeutic methods and compare different studies. Therefore, it seemed appropriate to study the effect of local treatment with different antibiotic carriers in the setting of an animal study. MATERIALS AND METHODS: The proven rat model for Staphylococcus aureus-induced osteomyelitis was used to compare the results of monotherapy with cefazolin, gentamicin-PMMA beads, or gentamicin-containing collagen sponge with the combination of local and systemic antibiotic treatment. RESULTS: Single-agent therapy with parenterally administered cefazolin reduced the CFU from 3.7 x 10(6) to 2.9 x 10(4) g(-1) of tibial bone. The effect on osteomyelitis was more pronounced with the local application of antibiotics. The best results were achieved with the gentamicin-containing collagen sponge which reduced the bacterial colony count to 1.4 x 10(2) CFU/g compared with 9.8 x 10(2) CFU/g achieved with gentamicin-PMMA beads. The effect was most marked using a 4-week combination therapy with local application of the gentamicin-containing collagen sponge and systemic administration of cefazolin. In 9 of 11 animals, no bacteria could be detected in the bone. CONCLUSION: Each of the treatment modalities resulted in a significant therapeutic effect. Due to its ability to quickly release large amounts of gentamicin, the flexible gentamicin-containing collagen sponge proved to be superior to the rigid PMMA system. Although the gentamicin-containing collagen sponge provided high antibiotic concentration at the site of implantation, an additive effect was attained when combined with systemic antibiotic treatment.
Asunto(s)
Antibacterianos/administración & dosificación , Cefazolina/administración & dosificación , Gentamicinas/administración & dosificación , Metilmetacrilatos/administración & dosificación , Osteomielitis/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológico , Animales , Colágeno/uso terapéutico , Desbridamiento , Modelos Animales de Enfermedad , Femenino , Osteomielitis/microbiología , Osteomielitis/cirugía , Vehículos Farmacéuticos/uso terapéutico , Polimetil Metacrilato/uso terapéutico , Ratas , Ratas Wistar , Infecciones Estafilocócicas/complicaciones , Staphylococcus aureusRESUMEN
This study evaluated the suitability of invA gene amplification by PCR as an effective means of detecting Salmonella species in pigs experimentally infected with S. Typhimurium DT104. A controlled infection study using 24 pigs was performed in order to compare efficacy, precision and detection rates of the invA-based PCR method originally described by Rahn, K. De Grandis, S.A., Clarke, R.C., McEwan, S.A., Galan, J.E., Ginocchio, C., Curtiss, R. 3rd, C.L. Gyles, (Mol. Cell. Probes 1992; 6: 271-279) as a new in-house invA-based PCR method for the specific detection of Salmonella spp. in pork and different tissue samples of slaughter pigs. Finally, PCR results were compared with culture detection rates obtained by isolation procedures following the ISO 6579:2000, the 'gold standard'. After slaughtering, 14 different tissue samples of each pig were investigated to verify the usefulness of the two invA-based PCR methods in different matrices of slaughter pigs. The results demonstrate that the application of the widely used invA-based primer pair (139 + 141) may result in questionable products if samples gained from selective enrichment in the Rappaport-Vassiliadis medium were investigated. These questionable products can lead to false-positive results, if no additional hybridization procedure is attached or if unspecialized persons use this method in routine laboratory practice. The newly developed in-house PCR method used is based on the 3'-prime region of invA, especially designed and harmonized for the detection of Salmonella in different matrices of slaughtered pigs after bacterial enriched broth culture. In this study, this PCR revealed no questionable products and, furthermore, the specificity of the amplificate could be tested by means of the restriction enzyme NdeI. In comparison with the culture detection procedure, the new PCR method has a sensitivity of 100% and a specificity of 96%. Thus, this method might be used as a meaningful tool in eliminating Salmonella-positive carcasses at slaughterhouse level and thus, keeping them out of the food chain.
Asunto(s)
Reacción en Cadena de la Polimerasa/veterinaria , Salmonelosis Animal/microbiología , Salmonella/patogenicidad , Enfermedades de los Porcinos/microbiología , Animales , Proteínas Bacterianas/genética , Técnicas de Tipificación Bacteriana/métodos , Técnicas de Tipificación Bacteriana/veterinaria , Cartilla de ADN , Masculino , Reacción en Cadena de la Polimerasa/métodos , Valor Predictivo de las Pruebas , Salmonella/clasificación , Salmonella/genética , Salmonelosis Animal/diagnóstico , Sensibilidad y Especificidad , Porcinos , Enfermedades de los Porcinos/diagnóstico , Factores de VirulenciaRESUMEN
In this study a Salmonella Typhimurium infection model in swine was used in order to investigate the influence of pre-mortal stress induced by long time period transportation on the re-activation of Salmonella in experimentally infected pigs. Salmonella free pigs were exposed to a highly virulent strain of Salmonella Typhimurium DT104 by direct intragastrical administration. Clinical parameters were monitored and the shedding rate in faeces was qualitatively and quantitatively determined by standard bacteriological procedures for 21 days. The distribution of the challenge organism in 14 different internal organs of transported and nontransported animals was determined. All infected animals developed clinical signs of salmonellosis 12 to 24 hours post infection. About 88 to 100% of the fecal samples were culture-positive up to post exposure day 6, and then varied from 71 to 92% until slaughter, respectively. At necropsy S. Typhimurium was recovered most frequently from caecum and ileocolic lymph nodes (83%), colon (79%), palatine tonsils (71%) and mandibular lymph nodes (62.5%). A negative impact of transportation stress on the shedding rate and the general condition of the animals was observed.
Asunto(s)
Salmonelosis Animal/etiología , Salmonella typhimurium/crecimiento & desarrollo , Estrés Fisiológico/veterinaria , Enfermedades de los Porcinos/etiología , Transportes , Animales , Heces/microbiología , Masculino , Salmonelosis Animal/prevención & control , Salmonella typhimurium/patogenicidad , Estrés Fisiológico/complicaciones , Porcinos , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/prevención & control , VirulenciaRESUMEN
The aim of this study was to investigate the suitability of the invA-based polymerase chain reaction (PCR) assay for the specific detection of Salmonella in organs of experimentally infected pigs and to compare these results to classical bacterial culture. While the PCR conditions specified in the "Deutsche Industrie Norm", DIN 10135 (section 35 LMBG, 1999), cutle based on the publication of Rahn et al. 1992, revealed various unspecific amplification products, modifications of the PCR conditions allowed the specific amplification of the invA fragment from inner organs. The modified PCR assay correlates exactly with cultivation results (as required by DIN Norm 6579) and enables the detection of Salmonella within 48 hours with equal sensitivity compared to routine cultivation.
Asunto(s)
Proteínas Bacterianas/genética , ADN Bacteriano/análisis , Salmonelosis Animal/diagnóstico , Salmonella typhimurium/aislamiento & purificación , Enfermedades de los Porcinos/diagnóstico , Animales , Secuencia de Bases , Genes Bacterianos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Salmonelosis Animal/microbiología , Salmonella typhimurium/clasificación , Salmonella typhimurium/genética , Sensibilidad y Especificidad , Porcinos , Enfermedades de los Porcinos/microbiologíaRESUMEN
Based on differences within the yopT-coding region of Yersinia. enterocolitica, Y pseudotuberculosis and Y pestis, a rapid and sensitive one-step polymerase chain reaction assay with high specificity for pathogenic Y enterocolitica was developed. By this method pathogenic isolates of Y enterocolitica can be easily identified and discriminated from other members of this genus. The entire coding sequence of the yopT effector gene of Y. pseudotuberculosis Y36 was determined.