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1

Genotypic markers of Yersinia enterocolitica O:9 isolated from cows positive in serological examination for bovine brucellosis

100%
Weiner M., Szulowski K., Iwaniak W., Zlotnicka J.
Bulletin of the Veterinary Institute in Puławy
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2012
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tom 56
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nr 4
The aim of the study was to perform a molecular investigations for the presence of pathogenicity genotypie markers of Y. enterocolitica 0:9 isolated from cattle, in which initially positive serological reactions for brucellosis were observed. Almost all isolates were αil-, ystA- and myfA-positive (n=19). On the other hand, one isolate, which harboured plasmid encoding gene yadA was αil-, ystA- and myfA-negative. The plasmid encoding yadA marker was present in half of the isolates tested. None of the examined isolates was ystß-positive. The results of the investigations revealed that the Y. enterocolitica O:9 isolates, related to false positive serological results for brucellosis, may be also potentially pathogenic for humans, due to the presence of chromosomal and plasmid- encoded molecular markers.
2

Development of a multiplex PCR for identification of Brucella sp. and cross-reacting Yersinia enterocolitica O:9

100%
Weiner M., Zlotnicka J., Iwaniak W., Szulowski K.
Bulletin of the Veterinary Institute in Puławy
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2011
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tom 55
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nr 4
The aim of the study was to develop a multiplex PCR, which allows an identification of the universal 16S rRNA Brucella sp. marker and amplification of the perosamine synthetase (per) gene, specific for cross-reacting Yersinia enterocolitica 0:9 only. The PCR analysis of the DNA extracted from all Brucella reference strains used in the investigations revealed the presence of the 16S rRNA gene, generating the amplicon of 905 bp size. Parallely, the examination of the DNA from Y. enterocolitica belonging to 0:9 serotype showed the presence of predicted amplicon of 312 bp typical to 0:9 serotype only. The sensitivity of the developed assay was determined with lymph tissue samples inoculated with B. abortus bvl and Y. enterocolitica 0:9 strains. The PCR analysis of the DNA extracted from lymph tissue revealed the presence of the predicted gene, when the 10⁶-10² CFU/g of the bacteria was added to the lymph tissue. The mPCR developed with direct extraction of DNA from lymph tissue can be a useful tool for the differentiation of infections caused by Brucella and cross-reacting Y enterocolitica 0:9.
3

Diagnosis of bovine brucellosis using traditional serological techniques and fluorescence polarisation assay

100%
Weiner M., Iwaniak W., Zlotnicka J., Szulowski K.
Bulletin of the Veterinary Institute in Puławy
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2010
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tom 54
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nr 4
The aim of this study was the application of fluorescence polarisation assay (FPA) for the examination of bovine sera and comparison of the results of the assay with the results of Rose Bengal test (RBT), serum agglutination test (SAT), complement fixation test (CFT), and ELISA. Six hundred thirty-five sera from cattle, including 300 sera from healthy animals, 32 sera from animals regarded as serologically positive for brucellosis and culled, and 303 sera originated from confirmatory investigations were used. All sera originating from healthy animals, negative in ELISA, RBT, SAT, and CFT were also negative in FPA. Among 303 sera from confirmatory investigations, 269 were positive in both RBT and SAT, 21 were positive in SAT and remaining 13 were RBT-positive only. Only two sera, one positive in both tests (RBT, SAT) and one SAT-positive, were also positive in FPA. Among 32 sera originated from animals regarded as serologically positive, which reacted in RBT, SAT, and CFT, 14 gave positive results in ELISA, whereas 18 were negative. Among these ELISA-positive sera, 13 were also positive in FPA. All samples positive in SAT, RBT, and CFT, and negative in ELISA, were also negative in FPA.
4
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Characteristics of Brucella strains isolated from animals in Poland

100%
Szulowski K., Iwaniak W., Weiner M., Zlotnicka J.
Polish Journal of Veterinary Sciences
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2013
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tom 16
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nr 4
A total of 42 Brucella strains were isolated from animals in Poland in years 2003-2012. Most of them (N=37) originated from wild animals, 3 from cattle, 1 from pig and 1 from sheep. The strains were characterised using both bacteriological and molecular (Bruce-ladder and MLVA) methods. The examinations revealed that all strains from wild boars, hares, cattle and pigs (N=41) had the same phenotypic characteristics and were classified as B. suis biovar 2. The remaining strain, isolated from sheep, was classified as B. ovis. The molecular examination showed that all B. suis biovar 2 strains, except one, had the same molecular profile as reference strain B. suis bv2 Thomsen. Different from the others strain originated from boars imported to Poland and its VNTR profile was typical for Iberian strains.
5
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Brucella suis biovar 2 isolations from cattle in Poland

100%
Szulowski K., Iwaniak W., Weiner M., Zlotnicka J.
Annals of Agricultural and Environmental Medicine
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2013
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tom 20
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nr 4
6

False positive serological reactions to brucellosis in pigs: a growing problem in international trade

76%
Szulowski K., Iwaniak W., Weiner M., Zlotnicka J., Szymajda M., Zareba Z., Czepinska H.
Medycyna Weterynaryjna
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2013
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tom 69
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nr 04
The diagnosis of brucellosis in pigs is based almost entirely on serological assays. None of the tests has been shown to be reliable in routine diagnosis in individual pigs. The biggest problem are false positive serological reactions (FPSRs) caused primarily by Yersinia enterocolitica O:9. The OPS component of the sLPS of Brucella is almost identical with that of Y. enterocolitica O:9. Thus no routinely used serological tests based on this antigen can distinguish between antibodies raised to these two infections. This paper presents the results of the examinations of 6 batches of pigs (total of 452 serum samples) traded between countries and causing major diagnostic problems. Positive reactions in RBT, SAT, CFT and I-ELISA were observed in all these batches of animals. Additionally 2 out of 21 samples from one of the batches were positive in 2-Me. FPSRs in the diagnosis of pigs for brucellosis seem to be a growing problem in international trade. The absence of provisions explicitly regulating the problem of FPSRs may have serious consequences, such as the slaughter of animals or even international repercussions. Clear guidelines for dealing with such cases should therefore be formulated.
7

Diagnostyka i sytuacja epidemiologiczna brucelozy bydła w Polsce

76%
Szulowski K., Iwaniak W., Weiner M., Zlotnicka J., Szymajda M., Zareba Z., Czepinska H.
Medycyna Weterynaryjna
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2012
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tom 68
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nr 02
The surveys of cattle for brucellosis in Poland are primarily based on serological tests. The examinations are performed by regional laboratories using RBT. In the case of positive results obtained in this test the samples are examined in SAT and CFT. The definitive confirmatory investigations are conducted by the National Reference Laboratory for Brucellosis in the Department of Microbiology of NVRI in Pu³awy, which additionally uses Coombs’ test, 2-Me test and ELISA. In the paper the results of the examination of cattle in Poland in the years 2005-2010 are shown. Each year during this period 1.1-1.3 million animals were included in the surveys and 130-317 cows were involved in confirmatory investigations. 12-34 animals were classified as positive for brucellosis. In bacteriological examinations of samples from seropositive cows, Brucella abortus was never once isolated. Since 2009 Poland is officially a brucellosis free country.
8

Survey of the anti-Brucella antibody status determined by ELISA testing in wild boars in Poland

76%
Szulowski K., Iwaniak W., Zlotnicka J., Szymajda M., Weiner M., Lipowski A., Jablonski A.
Medycyna Weterynaryjna
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2015
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tom 71
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nr 04
Sera of 4407 wild boars (Sus scrofa) shot by hunters in Poland in 2012 were tested by ELISA for antiBrucella antibodies. Samples originated from 11 out of 16 voivodeships, and 1077 (24.44%) seroreagents were detected. The highest prevalence of wild boars with anti-Brucella antibodies was found in Opolskie (39.9%) and Wielkopolskie (37.29%) voivodeships. The lowest percentage of positive results was observed in Kujawsko- -Pomorskie (13.74%) and Łódzkie (15.47%) voivodeships. The results for particular districts revealed significant differences in the prevalence of anti-Brucella antibodies, which varied between 0 and 100%. The results of the surveys show that wild boars constitute an enormous reservoir of Brucella suis biovar 2 microorganisms in Poland. The next stage of the study will see examinations of wild boars continued by bacteriological and molecular methods.
9
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Diagnostyka i sytuacja epidemiologiczna brucelozy świń w Polsce

76%
Szulowski K., Iwaniak W., Weiner M., Zlotnicka J., Szymajda M., Zareba Z., Czepinska H.
Życie Weterynaryjne
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2011
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tom 86
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nr 05
10

International trade - a potential source of brucellosis in pigs

76%
Szulowski K., Iwaniak W., Zlotnicka J., Weiner M., Zareba Z., Czepinska H.
Medycyna Weterynaryjna
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2011
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tom 67
|
nr 01
Brucellosis was confirmed in boars imported for breeding purposes to Poland from one of the EU countries. In serological investigations, in which the RBT, ELISA, CFT, SAT and 2-ME were used, positive reactions to brucellosis were found in 9 out of 23 animals. All the animals were slaughtered, and bacteriological examination was performed. Brucellae were isolated from the tissues of 7 boars. The characteristics of the strains isolated showed that they belonged to Brucella suis biovar 2, which is typical for Europe. The examination of animals for brucellosis at quarantine stations seems to be crucial for the protection of herds from Brucella infection.
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