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1.
Vet Microbiol ; 160(3-4): 378-86, 2012 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-22763172

RESUMO

Porcine brucellosis is a zoonotic disease of truly global significance because even in countries without the disease the occurrence of false positive serological reactions (FPSRs) creates significant problems. Statutory diagnostic testing is required in many disease free countries or regions and is often a prerequisite for the movement of live animals. Currently this testing is dependent almost entirely on serological assays and these may result in a significant number of FPSRs. The aim of this study was to examine existing and novel serodiagnostic assays to evaluate their diagnostic sensitivity and resilience to FPSRs. The existing assays evaluated were the RBT, smooth lipopolysaccharide (sLPS) indirect (i) ELISA, sLPS competitive (c) ELISA, and the FPA. The novel assays evaluated were the sLPS TR-FRET assay, a rough (r) LPS iELISA, a recombinant protein BP26 iELISA and a cytoplasmic protein extract (Brucellergene™) iELISA. Four populations of sera were evaluated: those from Brucella suis infected swine (n=34), randomly selected samples from non-infected swine (n=161), sera from non-infected swine within herds exhibiting FPSRs (n=132) and sera from swine experimentally infected with Yersinia enterocolitica O:9 (n=4). The results show that all the assays dependent on the sLPS O-polysaccharide (OPS) for their sensitivity (the RBT, sLPS ELISAs, FPA and the sLPS TR-FRET) had significantly reduced diagnostic specificity when applied to the FPSR population, the RBT being most affected. Of the two rapid homogeneous assays, the TR-FRET was diagnostically superior to the FPA in this study. Neither of the protein based iELISAs demonstrated sufficient diagnostic sensitivity to resolve the FPSRs. The rLPS iELISA showed no cross reaction with the FPSRs and had diagnostic sensitivity similar to that of the OPS based assays.


Assuntos
Brucelose/veterinária , Testes Sorológicos/veterinária , Animais , Anticorpos Antibacterianos/sangue , Brucella suis , Brucelose/diagnóstico , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática/normas , Ensaio de Imunoadsorção Enzimática/veterinária , Reações Falso-Positivas , Sensibilidade e Especificidade , Testes Sorológicos/métodos , Testes Sorológicos/normas , Suínos , Yersiniose/diagnóstico , Yersiniose/imunologia , Yersinia enterocolitica/imunologia
2.
J S Afr Vet Assoc ; 82(1): 56-7, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21826840

RESUMO

A preliminary serological study of 366 household dogs in Lagos and Ibadan, southwestern Nigeria, was carried out to determine antibodies due to exposure to Brucella abortus and B. canis, using the rose bengal test (RBT) and the rapid slide agglutination (RSA) test, respectively. Results showed that 5.46 % (20/366) and 0.27 % (1/366) of the dogs screened were seropositive to B. abortus and B. canis, respectively. Of all dogs, 36 had a history of being fed foetuses from cows and 11 (30.6 %) of these tested positive in the RBT. Our findings, although based on a limited sample size and a dearth of clinical details, revealed that dogs in Nigeria may be infected with Brucella spp. given the wide range of risk factors. Further studies are recommended to elucidate the epidemiology of brucellosis in dogs and its possible zoonotic consequences in the country.


Assuntos
Brucella abortus , Brucella canis , Brucelose/veterinária , Doenças do Cão/epidemiologia , Animais , Brucelose/epidemiologia , Brucelose/parasitologia , Cães , Nigéria/epidemiologia , Estudos Soroepidemiológicos
3.
Vet Rec ; 169(1): 14, 2011 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-21676987

RESUMO

Eight bottlenose dolphins (Tursiops truncatus) that stranded in Cornwall, south-west England, between June 2004 and December 2007 were examined using standardised postmortem examination and bacteriological methods. Evidence of Brucella species infection was found in four of these dolphins on culture. In addition, of the eight dolphins, four were positive and two were weakly positive for antibodies to Brucella species on serological analyses of pericardial and other fluids using a competitive ELISA and two indirect ELISAs. High or very high levels of the sum of 25 individual chlorobiphenyl congeners (∑25CBs) were also determined in blubber samples from two of the dolphins (45.5 and 446.6 mg/kg lipid weight).


Assuntos
Tecido Adiposo/química , Golfinho Nariz-de-Garrafa , Brucelose/veterinária , Bifenilos Policlorados/metabolismo , Animais , Golfinho Nariz-de-Garrafa/metabolismo , Golfinho Nariz-de-Garrafa/microbiologia , Brucelose/epidemiologia , Inglaterra/epidemiologia , Poluentes Ambientais , Feminino , Masculino
5.
J Wildl Dis ; 44(2): 237-46, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18436657

RESUMO

Adult female nematodes identified as Pseudalius inflexus were collected from the lungs of a juvenile male harbor porpoise (Phocoena phocoena) found dead on a beach in Cornwall, UK. Classic and molecular typing methods, immunologic and electron microscopy immunolabeling techniques, provided evidence of Brucella sp. infection within the uterine tissue of nematodes of this marine mammal. This finding presents further evidence to suggest parasites should be considered as a potential means of transfer of bacterial infection in marine mammals and highlights the zoonotic implications for humans exposed to marine mammals through occupation or leisure.


Assuntos
Brucella/isolamento & purificação , Nematoides/microbiologia , Toninhas/parasitologia , Animais , Brucella/patogenicidade , Impressões Digitais de DNA , DNA Bacteriano/análise , Evolução Fatal , Feminino , Pulmão/parasitologia , Masculino , Microscopia Eletrônica de Transmissão/métodos , Microscopia Eletrônica de Transmissão/veterinária , Nematoides/ultraestrutura
6.
J Clin Microbiol ; 44(12): 4363-70, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17035490

RESUMO

Naturally acquired infection of humans with a marine mammal-associated Brucella sp. has only been reported once previously in a study describing infections of two patients from Peru. We report the isolation and characterization of a strain of Brucella from a New Zealand patient that appears most closely related to strains previously identified from marine mammals. The isolate was preliminarily identified as Brucella suis using conventional bacteriological tests in our laboratory. However, the results profile was not an exact match, and the isolate was forwarded to four international reference laboratories for further identification. The reference laboratories identified the isolate as either B. suis or B. melitensis by traditional bacteriological methods in three laboratories and by a molecular test in the fourth laboratory. Molecular characterization by PCR, PCR-restriction fragment length polymorphism, and DNA sequencing of the bp26 gene; IS711; the omp genes omp25, omp31, omp2a, and omp2b; IRS-PCR fragments I, III, and IV; and five housekeeping gene fragments was conducted to resolve the discrepant identification of the isolate. The isolate was identified to be closely related to a Brucella sp. originating from a United States bottlenose dolphin (Tursiops truncatus) and common seals (Phoca vitulina).


Assuntos
Brucella/classificação , Brucella/isolamento & purificação , Brucelose/microbiologia , Osteomielite/microbiologia , Doenças da Coluna Vertebral/microbiologia , Animais , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Golfinho Nariz-de-Garrafa/microbiologia , Brucella/genética , Brucella/fisiologia , Análise por Conglomerados , Elementos de DNA Transponíveis , DNA Bacteriano/química , DNA Bacteriano/genética , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Nova Zelândia , Phoca/microbiologia , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA
8.
Rev Sci Tech ; 25(3): 1039-53, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17361769

RESUMO

The principal methods for the serological diagnosis of bovine brucellosis are the complement fixation test (CFT), serum agglutination test (SAT), Rose-Bengal test (RBT), indirect enzyme-linked immunosorbent assay (iELISA) and more recently the competitive ELISA (cELISA) and the fluorescent polarisation assay (FPA). Guidelines set by the World Organisation for Animal Health (OIE) describe methods and diagnostic thresholds for each of these tests. Many countries have adopted these methods for the purposes of eradication of brucellosis and have legislated for the use of these tests (the CFT and SAT in particular) for the prevention of the spread of the disease through international trade. Within the European Union (EU) each member state has a National Reference Laboratory which regulates the quality of brucellosis diagnosis and works to the recommendations set by the OIE. This article describes the results from the first three EU ring trials assessing the harmonisation of diagnostic tests between each member state. The general level of harmony for SAT, CFT, and iELISA was found to be good, but issues of standardisation of the RBT, cELISA and FPA remain. The cELISA and FPA in particular need further work to create European harmony. The ring trials also proved successful at providing specific evidence of poor performance in some areas. The decision on whether or not to take action on the basis of these results rested with the individual laboratories concerned. The increase in the number of participants in these trials over time reflected the enlargement of the EU and increased the need for quality assurance.


Assuntos
Brucelose Bovina/diagnóstico , Técnicas de Laboratório Clínico/veterinária , Kit de Reagentes para Diagnóstico/veterinária , Animais , Bovinos , Técnicas de Laboratório Clínico/normas , Diagnóstico Diferencial , União Europeia , Controle de Qualidade , Kit de Reagentes para Diagnóstico/normas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
12.
J Immunol Methods ; 278(1-2): 171-8, 2003 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12957405

RESUMO

The fluorescence polarisation assay (FPA) is a recently described test for the serological diagnosis of Brucella infection. It has many methodological advantages over older, more established tests and can be performed in a fraction of the time. To validate the FPA, serum samples from 146 confirmed (by culture) Brucella-infected cattle were tested in conjunction with serum samples from 1947 noninfected cattle. The competitive ELISA (cELISA) was validated using these positive reference samples and 1440 negative samples, while data for the indirect ELISA (iELISA) was generated from 6957 negative samples plus the positive sera. Published diagnostic specificity (DSp) data for the complement fixation test (CFT) and serum agglutination test (SAT) was used in conjunction with the test results on the positive sera to obtain diagnostic specificity plus diagnostic sensitivity (DSn). After selection of a cutoff for the FPA and cELISA, the diagnostic specificity and sensitivity total for each test were compared. The results, with 95% confidence intervals, were: FPA (195.7+/-2.79), iELISA (195.0+/-2.70), cELISA (194.9+/-3.48), CFT (191.7+/-4.45), and SAT (180.4+/-6.33). The data presented supports the use of the FPA in diagnosis of brucellosis and questions the use of the SAT and CFT for either screening or confirmatory testing.


Assuntos
Brucella abortus/imunologia , Brucelose Bovina/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Imunoensaio de Fluorescência por Polarização/veterinária , Testes de Aglutinação/veterinária , Animais , Brucelose Bovina/diagnóstico , Bovinos , Testes de Fixação de Complemento/veterinária , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
13.
J Appl Microbiol ; 92(4): 724-8, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11966913

RESUMO

AIMS: To select an anti-fungal agent to replace cycloheximide in the media used for isolation of Brucella. METHODS AND RESULTS: One potential agent, natamycin, was evaluated using 28 Brucella isolates, 18 yeasts and 14 fungi. The material for the evaluation included 37 bovine milk samples, six bovine vaginal swabs and 45 milk samples artificially infected with Brucella. The recovery of Brucella only from the artificially-inoculated milk samples increased with the use of the medium containing natamycin instead of cycloheximide, at the same time significantly inhibiting the growth of yeasts, fungi and other bacteria. The inclusion of either anti-fungal agent allowed growth of the 28 Brucella isolates and totally prevented the growth of all 18 yeasts and 13 of the 14 fungi. CONCLUSIONS: Based on the results it was concluded that natamycin would be a suitable alternative to cycloheximide. SIGNIFICANCE AND IMPACT OF THE STUDY: Cycloheximide has become unavailable worldwide and is currently an anti-fungal constituent of the medium often used for isolation of Brucella organisms. The use of natamycin as a replacement in the formulation did not inhibit growth of Brucella and was effective at eliminating most contaminants.


Assuntos
Antifúngicos/farmacologia , Brucella/crescimento & desenvolvimento , Brucella/isolamento & purificação , Fungos/efeitos dos fármacos , Natamicina/farmacologia , Animais , Técnicas Bacteriológicas , Brucelose Bovina/microbiologia , Bovinos , Meios de Cultura , Cicloeximida/farmacologia , Feminino , Leite/microbiologia , Vagina/microbiologia
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