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1

Evaluation of the ELISA in diagnosis of bovine brucellosis. Part II - Examination of milk

100%
Szulowski K.
Polish Journal of Veterinary Sciences
|
1999
|
tom 02
|
nr 2
In a previous paper an ELISA for detecting anti-Brucella antibodies in cattle sera was elaborated and evaluated. The aim of this study was to evaluate the ELISA for testing individual milk samples from cattle. Validation was performed by comparing the results of this test with those in the Rose Bengal Test (RBT), tube agglutination test (TAT), complement fixation test (CFT), Coombs anti-globulin test (Coombs), 2-mercaptoet- hanol test (2-ME) and serum ELISA. The examinations involved 79 milk samples from cows in which sera were positive in the RBT, 14 milk samples from cows in which sera were positive in the CFT, Coombs or 2-ME, 530 milk samples from cows considered free of brucellosis and 309 milk samples from cows from herds suspected of brucellosis. No correlation between the milk ELISA and RBT results was found - among 79 samples from cows in which sera reacted positively in the RBT only 22 were positive or doubtful in the ELISA, but all samples in which the presence of anti-Brucella antibodies was confirmed in the CFT, Coombs or/and 2-ME reacted positively in the ELISA (14 out of 14 samples). All 530 milk samples from cows free of brucellosis were negative in the ELISA. Among 309 milk samples from cows from herds suspected of brucellosis 11 positive and 20 doubtful results were obtained in the ELISA and this method proved to be more sensitive than traditional methods and serum ELISA, in which method only 3 samples from the same cows reacted positively and 2 doubtfully. The ELISA for testing individual milk samples proved to be a useful method for diagnosis of brucellosis in cattle.
2
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Application of meat juice in diagnosis of brucellosis in hares and wild boars by ELISA

100%
Szulowski K., Pilaszek J., Iwaniak W.
Bulletin of the Veterinary Institute in Puławy
|
2000
|
tom 44
|
nr 1
Meat juice and serum samples from hares and wild boars were used in the ELISA for the examination for the presence of anti-Brucella antibodies. A high correlation between meat juice ELISA and serum ELISA was obtained. All 9 meat juice samples from hares with anatomopathological lesions characteristic of brucellosis (from 8 animals Brucella was isolated) were positive in the ELISA, similarly as sera. Out of 46 remaining hares only 1 reacted positively in the meat juice ELISA and it was not confirmed by the serum ELISA and other methods. As concerns the material from wild boars 12.3% of sera (out of 933 samples) and 13.7% of meat juice samples ( out of 336) were positive in the ELISA. Among 48 serum and meat juice samples coming from the same animals, 6 samples were positive both in the serum ELISA and meat juice ELISA and 42 were negative. The meat juice ELISA proved to be a valuable method for testing both hares and wild boars for anti-Brucella antibodies.
3

Using combined recombinant protein in the diagnosis of bovine brucellosis

84%
Bulashev A., Jakubowski T., Mukantayev K., Tursunov K., Kiyan V., Zhumalin A.
Medycyna Weterynaryjna
|
2018
|
tom 74
|
nr 03
The aim of this research was to obtain a combined recombinant protein consisting of immunodominant regions of Brucella outer membrane proteins (Omps) with the molecular weights of 25 kDa (Omp25) and 31 kDa (Omp31). The search for candidate proteins was carried out using NCBI PubMed and NCBI GenBank databases. The two most immunodominant regions of Omps were selected. The first region was in a position from 48 to 83 amino acids of Omp31, while the second one was in the position from 180 to 224 amino acids of Omp25. This combined sequence was designated as OmpBm-Ba. The pET32 vector was used for cloning and the expression of the combined recombinant protein in E. coli BL21(DE3). The antigenicity of recombinant OmpBm-Ba (rOmpBm-Ba) as compared with rOmp25 and rOmp31 has been tested on the sera of 109 cattle with positive results to brucellosis according to classical serological tests. The rOmpBm-Ba had a higher antigenicity than the single ones, since it confirmed the presence of Brucella-antibody in all serum samples with positive results of i-ELISA/rOmp25 and/or i-ELISA/rOmp31, as well as additionally revealing 7.3% and 31.2% seropositive animals, respectively. A comparative study of the diagnostic value of i-ELISA/rOmpBm-Ba and conventional tests on blood sera of 24 cattle subjected to a bacteriological examination at slaughter showed a higher reliability of enzyme immunoassay. Further research is necessary to get a more objective evaluation of the diagnostic accuracy of Brucella rOmpBm-Ba by challenging laboratory animals.
4

Evaluation of in vitro activities of tigecycline and various antibiotics against Brucella spp.

84%
Ozhak-Baysan B., Ongut G., Ogunc D., Gunseren F., Sepin-Ozen N., Ozturk F., Aktepe O. C., Gultekin M.
Polish Journal of Microbiology
|
2010
|
tom 59
|
nr 1
55-60
Brucellosis is a zoonosis with a worldwide distribution and remains a significant public health problem mainly in the developing world. In this study we evaluated the in vitro activities and synergistic effects of antibiotic combinations against blood culture isolates of Brucella spp. In vitro susceptibilities of 76 blood culture isolates of Brucella melitensis and one blood culture isolate of Brucella abortus to doxycycline, streptomycin, gentamicin, trimethoprim-sulfamethoxazole, moxifloxacin, rifampin, ciprofloxacin, and tigecycline were examined by Etest method. For 37 patients with Brucella spp. isolates (36 B. melitensis, 1 B. abortus), antibiotic combinations used for treatment were identified with those tested in vitro for synergy using Etest method. Trimethoprim-sulfamethoxazole and tigecycline were the most active of the compounds tested with MIC₉₀ value of 0.094 mg/1. Among antibiotic combinations only streptomycin-rifampin combination was synergistic for one Brucella spp. isolate. The other antibiotic combinations revealed antagonistic or indifferent activity. Complete clinical response was achieved in all patients. Further studies are required to determine the correlation between the antimicrobial susceptibility and synergy test results with the clinical course of patients. Brucellosis can be adequately treated with existing regimens in our region.
5

Occurrence of Yersinia enterocolitica O:9 in feces from cows seropositive for brucellosis

84%
Szulowski K., Weiner M., Iwaniak W.
Medycyna Weterynaryjna
|
2014
|
tom 70
|
nr 01
The diagnosis of brucellosis is mainly based on serological tests. All animals classified as serologically positive are obligatorily slaughtered and subjected to bacteriological examination. B. abortus has not been reported in Poland since the eradication of bovine brucellosis in 1980. On the other hand, B. suis biovar 2 is sporadically isolated from cattle. In accordance with the instructions of the Chief Veterinary Officer, samples of feces from animals slaughtered following positive serological results for brucellosis have been examined for the presence of Yersinia enterocolitica O:9 since 2011. Because of the similarity of its O-polysaccharide component of S-LPS with that of Brucella, this microorganism is considered to be a major contributory factor of false positive serological reactions (FPSR). The paper presents the results of the bacteriological and molecular examination of feces from 26 cows seropositive for brucellosis and 30 healthy cows, negative for brucellosis, for the presence of Y. enterocolitica O:9. Y. enterocolitica O:9 was found in 7 of the positive cows, whereas all samples from cows negative for brucellosis were free from this bacteria. These results indicate that Y. enterocolitica O:9 may be responsible for some of the positive results for brucellosis in cattle.
6

Evaluation of real-time PCR method for rapid diagnosis of brucellosis with different clinical manifestations

84%
Surucuoglu S., El S., Ural S., Gazi H., Kurutepe S., Taskiran P., Yurtsever S. G.
Polish Journal of Microbiology
|
2009
|
tom 58
|
nr 1
15-19
In this study, we tested the advantages of TaqMan real time PCR technique and compare it to conventional methods using serum samples from patients with different clinical forms of brucellosis. A total of 50 patients were included in the study. Blood culture using BACTEC 9240 system, Standard Wright's tube agglutination, and real time PCR methods were used. Control blood samples from 30 people with no history of brucellosis or exposure to Brucella spp. were examined too. Serological assay was positive for 49 patients (98%). Forty-four (88%) of the 50 patients had a positive PCR result, whereas Brucella spp were isolated from blood cultures of 18 patients (36%). STA test was all positive for focal brucellosis. Real time PCR test was positive in 9 patients with focal disease (90%), whereas blood culture was positive only in 4 patients (40%). The sensitivity, specificity, positive and negative predictive values of the real time PCR method were calculated as 88%, 100%, 100%, and 83%, respectively. Our results suggest that the high sensitivity and specificity of real time PCR method make it a useful tool for diagnosis of brucellosis with different clinical manifestations.
7

Expression of human serum albumin - L7-L12 [Brucella abortus ribosomal protein] fusion protein in Saccharomyces cerevisiae

84%
Pakzad I., Rezaee A., Emaneini M., Hosseini A. Z., Tabbaraee B., Taherikalani M.
Polish Journal of Microbiology
|
2009
|
tom 58
|
nr 2
99-104
Brucella abortus is a facultative intracellular gram-negative bacterial pathogen that causes abortion in pregnant cattle and undulant fever in humans. The immunogenic B. abortus ribosomal protein L7/L12 is a promising candidate antigen for the development of subunit vaccines against brucellosis. It has already been expressed in several bacteria and has been used as DNA vaccine. In order to construct yeast expressing vector for the tHSA-L7/L12 fusion protein, the 171112 ribosomal gene was amplified by PCR. The expression plasmid pYtHSA-L7/Ll 2 was constructed by inserting the L7/L12 gene into the pYHSA5 shuttle vector (containing inulinase signal sequence, HSA gene and Gal 10 promoter). The recombinant vector was transformed into S. cerevisiae and was then induced by galactose. The secreted recombinant fusion protein was detected in supernatant by SDS-PAGE and confirmed by western blot analysis using anti-HSA and anti-L7/L 12 antibodies. Fusion protein was purified by affinity chromatography and its amount was approximately 500 μg/liter.
8

Isolation and biotyping of Brucella melitensis from aborted sheep foetuses in Turkey

84%
Sahin M., Unver A., Otlu S.
Bulletin of the Veterinary Institute in Puławy
|
2008
|
tom 52
|
nr 1
Aborted sheep foetuses were collected during the lambing seasons of 2004 to 2006. B. melitensis was isolated from 25 (29.76%) of 84 lungs and stomach contents. Based on the biochemical tests and agglutination test with monospecific A and M antisera, the isolates were determined as B. melitensis biotype 3. The isolation of the B. melitensis from aborted sheep foetuses may show the importance of this agent in the aetiology of ovine brucellosis in Turkey. The determination of dominant biotype of B. melitensis responsible for ovine brucellosis is also expected to help to understand the epidemiology of ovine brucellosis in detail in this region and neighbouring countries.
9

Comparison of rose bengal plate test antigens prepared from Brucella abortus, Brucella melitensis and Brucella suis

84%
Erganis O., Hadimli H. H., Solmaz H., Corlu M.
Bulletin of the Veterinary Institute in Puławy
|
2005
|
tom 49
|
nr 2
10
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Porcine brucellosis in Poland: problems accompanying serological surveys conducted in 1995-2000

84%
Szulowski K., Iwaniak W., Pilaszek J.
Bulletin of the Veterinary Institute in Puławy
|
2001
|
tom 45
|
nr 2
Analysis of problems relevant to serological surveys of pigs for brucellosis was the aim of the studies. In these investigations 74 218 sera from domestic pigs monitored for brucellosis and 3 566 sera from imported pigs were used. For comparison study 159 pig serum samples, collected from three Brucella infected herds, were tested, using following methods: RBT, ELISA, CFT, TAT and 2-ME. The obtained results indicate that serological diagnosis of brucellosis in domestic pigs did not produce major problems. Different situation was observed when some batches of imported pigs were examined. The results of serological investigations were characterised by disquietingly high proportion of samples positive in the RBT, TAT, CFT or ELISA. The analysis of documents and epidemiological inquiry concerning the pigs revealed that they originated from herds struggling with a problem of false positive serological reactions (FPSR), probably due to Yersinia enterocolitica 0:9 infection.
11

Brucellosis in Mirpur, Azad Kashmir Pakistan: a livestock threat for neighboring zones

67%
Mubeen H., Khan I., Saleem M. H., Akhtar R., Ali S., Saqib M., Khan A. U., Umar S.
Medycyna Weterynaryjna
|
2018
|
tom 74
|
nr 07
The objective of the present study was to determine the prevalence of brucellosis in household animals of Mirpur, Azad Kashmir due to its geographic importance. A total of 360 blood samples of cattle, buffaloes, sheep and goats were initially screened through Rose Bengal Plate test (RBPT) and then positive samples were subjected to Enzyme Linked Immunosorbent Assay (ELISA) for confirmation and quantification of antibody titers. Molecular confirmation of serologically positive samples was performed by real time polymerase chain reaction (PCR). RBPT and ELISA showed a total of 8.6% and 6.87% positive samples respectively. The species wise seropositivity by RBPT was greater in cattle followed by buffaloes, goats and sheep. Similarly ELISA showed more seropositivity in cattle than buffaloes, while sheep and goats were negative for brucellosis by ELISA. RT-PCR revealed 100% samples positive for Brucella abortus by species specific PCR. This study revealed the presence of Brucella abortus in Mirpur for the first time. Since brucellosis is listed in transboundary diseases, its presence in this geographically important region could be a potential threat for neighboring countries.
12

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

67%
Weiner M., Iwaniak W., Zlotnicka J., Szulowski K.
Bulletin of the Veterinary Institute in Puławy
|
2010
|
tom 54
|
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.
13

Comparison of different PCR methods for detection of Brucella spp. in human blood samples

67%
Al-Ajlan H., Ibrahim A., Al-Salamah A.
Polish Journal of Microbiology
|
2011
|
tom 60
|
nr 1
For detection of Brucella species by PCR four DNA extraction methods and four targets were compared using pure culture of Brucella melitensis and the best conditions were applied in clinical samples. It was found that the MagNA Pure LC method was the most efficient and sensitivemethod showing a positive PCR reaction with DNA extracted from as low as 25 and 100 CFU suspended in one ml blood and one ml water, respectively. Detection of Brucella spp. by conventional PCR was investigated using four different targets. The results indicated that The B4-B5 amplification method was the most sensitive one as it could amplify DNA extracted from as a low as 25 and 100 CFU/ml suspended in one ml water and blood, respectively. Furthermore real-time PCR was able to detect Brucella using DNA extracted from as low as 50 CFU/ml blood and 15 CFU/ml water, respectively. The best and optimum detection conditions were applied to the clinical samples. Evaluation of conventional PCR assays on blood specimens confirmed 72% of the results obtained by conventional blood culture methods with a specificity of 95%, while serum samples had a sensitivity of 54% and specificity of 100%. Real-time PCR was generally found to be more sensitive and specific for detecting Brucella spp. in blood and serum samples compared to conventional PCR. The real-time PCR done on blood specimens confirmed 77.5% of the results obtained by conventional blood culture methods with specificity of 100%, while 60% of serum samples were found to be positive with specificity of 100%. These results suggest that serum and blood analysis by conventional and real time PCR is a convenient and safe method for rapid and accurate diagnosis of brucellosis.
14

Detecting Toxoplasma, Listeria and Brucella antibodies in goitered gazelles in Turkey

67%
Gokcen A., Altas M. G., Sevgili M., Babur C., Celebi B., Kilic S.
Medycyna Weterynaryjna
|
2007
|
tom 63
|
nr 09
1064-1066
The aim of this study was to determine the prevalence of toxoplasmosis, listeriosis and brucellosis in goitered gazelles (Gazella subgutturosa) in Sanliurfa region, Turkey. A total of 82 sera were collected from healthy gazelles and tested for listeriosis, brucellosis and toxoplasmosis by the Osebold Agglutination Test (OAT), Serum Agglutination Test (SAT) and Sabin-Feldman Dye Test (SFDT), respectively. 82 gazelles 5 (6.09%) were seropositive for listeriosis, 23 (28.04 %) for toxoplasmosis and all of them were seronegative for brucellosis. No statistically significant differences were observed between male and female gazelles in the seroprevalences of toxoplasmosis and listeriosis. As a result, the presence of anti-Toxoplasma gondii and Listeria spp. specific antibodies in G. subgutturosa in the region of Sanliurfa was determined.
15

International trade - a potential source of brucellosis in pigs

67%
Szulowski K., Iwaniak W., Zlotnicka J., Weiner M., Zareba Z., Czepinska H.
Medycyna Weterynaryjna
|
2011
|
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.
16
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Seroprevalence of Q fever, brucellosis and leptospirosis in farmers and agricultural workers in Bari, Southern Italy

67%
Monno R., Fumarola L., Trerotoli P., Cavone D., Giannelli G., Rizzo C., Ciceroni L., Musti M.
Annals of Agricultural and Environmental Medicine
|
2009
|
tom 16
|
nr 2
205-209
17

Evaluation of the ELISA in diagnosis of bovine brucellosis. Part I - Examination of sera

67%
Szulowski K.
Polish Journal of Veterinary Sciences
|
1998
|
tom 01
|
nr 2
The investigation aimed at preparing an ELISA kit for examination of bovine sera for brucellosis and evaluating the diagnostic properties of the test. Lipopolysaccharide extract (LPS) was used as the antigen for microplate coating, antibodies against IgG of bovine sera with horseradish peroxidase were used as the conjugate and ABTS with H202 as the substrate. A weak-positive serum prepared from the second International Standard of anti-Brucella abortus Serum (ISaBaS II) and a negative serum obtained by mixing sera from Brucella-free cows were used as controls. Evaluation of the diagnostic usefulness of the ELISA was performed by comparison of the results obtained with those in the rose bengal test (RBT) and complement fixation test (CFT). The examinations involved 212 bovine sera positive in the RBT, 60 sera positive in the CFT, 5 standard sera, and 1005 sera negative in the RBT. A high correlation was found between the ELISA and CFT results (55 sera out 60 sera) and no correlation between the ELISA and RBT results. Only 60 out of 212 sera positive in RBT gave positive results in the ELISA. All standard sera were positive in the ELISA and only 1 serum from those negative in the RBT was doubtful in the ELISA. The ELISA proved to be a suitable method for diagnosis of brucellosis in cattle.
18
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Brucellosis in humans - etiology, diagnostics, clinical forms

59%
Galinska E. M., Zagorski J.
Annals of Agricultural and Environmental Medicine
|
2013
|
tom 20
|
nr 2
Brucellosis in humans is a zoonosis of greatly varied clinical image. It occurs on all inhabited continents. The course of the disease may be acute, sub-acute or chronic. The etiologic factors of brucellosis are small, aerobic Gram-negative rods of the genus Brucella, which currently contains ten species: B. abortus, B. suis, B. ovis, B. melitensis, B. canis, B. neotomae, B. pinnipedialis, B. ceti, B. microti and B. inopinata. In humans, the disease is caused mainly by: B. melitensis as the most pathogenic species, followed by B. suis, whereas B. abortus is considered as the mildest type of brucellosis. The natural reservoir of the germ and the source of infection in humans are infected domestic animals, primarily cattle, sheep, goats, as well as wild animals. Infection in humans occurs by penetration through damaged skin, conjunctiva, and more rarely via the alimentary route by the consumption of infected products. Especially exposed are: veterinarians, veterinary technicians, insemination service employees, zoo technicians, farmers working on multi-herd farms (production cooperatives), e.g. cattlemen, also private farmers, employees of slaughter houses and meat processing enterprises. A basis for diagnosing brucellosis are serologic tests which allow the detection of antibodies occurring in response to infection, performed with the use of the following methods: agglutination test, complement fixation test, Coombs test, 2-mercaptoethanol agglutination test, and Burnet’s intradermal allergy test which detects the state of hypersensitivity of the infected organism to Brucella abortus rods.
19

Najważniejsze zakaźne przyczyny ronień u bydła

43%
Ptaszynska M.
Lecznica Dużych Zwierząt. Ogólnopolski Kwartalnik dla Lekarzy Weterynarii
|
2015
|
tom 10
|
nr 2 Monografia
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