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    Isolation, Identification and Typing of Brucella Species as Zoonotic Pathogens by Using Conventional and Molecular Biological Methods (2010)

    Art
    Hochschulschrift
    Autor
    Gwida, M.
    Quelle
    Berlin: Mensch und Buch Verl, 2010 — 96 Seiten
    ISBN: 978-3-86664-854-8
    Kontakt
    Institut für Tier- und Umwelthygiene

    Robert-von-Ostertag-Str. 7-13
    Gebäude 35
    14169 Berlin
    +49 30 838 51845
    tierhygiene@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    Representative studies from countries including North Africa, the Middle East, and India which are neighbours or are important trading partners of the European Union and on trade animals and their product. In our review, published data on seroprevalence of brucellosis from 1948 to 2009 were retrieved. Based on the collected literature, we found that new seroprevalence data are needed urgently to evaluate the current situation and for continuous monitoring of necessary control programs.
    Sera of 895 apparently healthy Camels (Camelus dromedaries) in addition to 5 human samples from Central Veterinary Research Laboratory (CVRL); Dubai, UAE was investigated. Rose Bengal Test (RBT), Complement Fixation Test (CFT), Slow Agglutination Test (SAT), Competitive Enzyme Linked Immunosorbant Assay (cELISA) and Fluorescence Polarization Assay (FPA) as well as real-time PCR were used. Our findings revealed that bcsp31 kDa real-time PCR detected Brucella DNA in 84.8% (759/895) of the examined samples, of which 15.5% (118/759) were serologically negative. Species specific real-time PCR system revealed that only Brucella abortus was present in the camels investigated in this study. A probit analysis revealed that real-time PCR assay detect as little as 23fg of Brucella DNA per reaction with a probability of 95%. Our results show no relevant difference in sensitivity between the different serological tests. FPA detected the highest number of positive cases (79.3%) followed by CFT (71.4%), RBT (70.7%), SAT (70.6%) and cELISA (68.8 %). A perfect agreement between CFT, RBT and SAT was proven by calculating Kappa values but sensitivity of all tests is low when compared to FPA or real-time PCR. A combination of real-time PCR with one of the used serological tests identified brucellosis in more than 99 % of the infected animals. 59.7% of the examined samples were positive in all serological tests and real-time PCR. On the other hand, Brucella spp was also isolated from animal handler. The percentage of positive cases among the examined samples were 60, 100, 80, 80,100 by using RBT, cELISA, CFT, SAT, FPA, respectively. On the other hand, real-time PCR detected infection among 40% from the examined samples and those samples were also positive in most of the used serological tests. Species specific real-time PCR revealed that the infection in human cases was also due to B. abortus. This result confirmed the hazards associated with occupational exposure through direct contact with infected animals.
    A total of 46 B. melitensis isolate obtained from sheep and human from Turkey were studied. From which, 20 were isolated from aborting ewes in different sheep flocks in Van Province, East Anatolia in Turkey during lambing seasons 2004-2005, 2005-2006 and 2006-2007, as well as 26 strains from German tourists were isolated in Germany and were sent to the National Reference Laboratory, were also included in this study. They were collected during 2004 to 2010. Our investigation was amended with recently published data for 41 B. melitensis isolates from German tourists (Al Dahouk et al., 2007b). The 67 human B. melitensis strains isolated in Germany and the 20 sheep isolates from Turkey, Southeast Anatolia were clustered in 68 different genotypes based on the differences in the numbers of repeat units at 16 VNTR loci. It is concluded that brucellosis is highly prevalent in sheep and humans in several district in Turkey.