Abort Yapan Sığırların Vajinal Sıvap Örneklerinden Brucella spp. Tespit Edilmesi: İmmunoperoksidaz ve Bakteriyolojik Kültür Yönteminin Karşılaştırılması (original) (raw)
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Brucellosis is an important disease in terms of economic losses in cattle breeding. Brucella spp. associated abortion is common in Kars, Turkey. Determination of Brucellosis cases therefore poses great importance. In this study, an immunochemical staining technique was tested in detecting cattle with Brucellosis and compared to bacteriological culture technique. Post-abortion, total of 261 vaginal swab samples were collected from cows. Brucella spp. were isolated from 25 animals by cultural method. All Brucella isolates were identified as Brucella abortus field strain and biotyped as B. abortus biotype 3. Smears prepared from culture positive vaginal samples were stained by immunoperoxidase technique, and 22 out of 25 animals were detected as Brucella-positive. Sensitivity and specificity of the immunoperoxidase technique were calculated as 88% and 92%, respectively.
Abortion is the most obvious manifestation of Brucella infection. In this study, 59 aborted buffalos and 91 aborted cows were included. Diagnosis of Brucella infection in these abortions was based on clinical, serological, bacteriological, hormonal, and molecular assays. Serological studies included the use of RB and ELISA tests as screening tests for infection. Argumentative differences between RB and ELISA results have been shown. Brucella isolated and identified from aborted fetuses, vaginal discharge and milk samples were 7 isolates from aborted cows and 3 from aborted buffalos. Brucella isolates revealed amplification of a 223-bp fragment with B4 and B5 primers. Hormonal assessment in both, brucella infected cows and buffalos, registered significant decrease in progesterone and P/E ratio in comparison with that aborted due to other causes. Immunohistochemical study revealed down expression level of 3BHSD enzyme in placentas of Brucella positive animals. ELISA technique was the valuable serological test to confirm the diagnosis of brucellosis. In conclusion, both RB and ELISA are necessary to be performed together as screening tests in diagnosis of brucellosis, whereas serum hormonal, placental immunohistochemical, and molecular (PCR) assessments have an efficient diagnostic values which can be included for confirmation of brucellosis.
Iranian journal of veterinary research, 2017
Rapid and precise diagnosis in natural field cases of bovine abortion caused by Brucella abortus warrants the use of the most sensitive and reliable diagnostic methods. In the present study, bacterial isolation, serology, gross, histopathology, immunohistochemistry and polymerase chain reaction technique(s) were applied. Sero-prevalence studies showed the rate of 28.86% positive cases using the competitive ELISA. Histopathological changes were mainly seen in the placenta, fetal lungs, kidney, liver and spleen. Immunohistochemical (IHC) staining of Brucella spp. was evident as brown, finely granular intracytoplasmic staining in trophoblasts of placental sections and in section(s) of liver, lung, kidney and spleen. Twenty-eight out of the 103 samples (17 from stomach contents, 3 from placental cotyledons, 2 from vaginal discharges and 6 from pooled fetal tissues) produced 193 bp amplicon specific for Brucella genus. Moreover, the species-specific primers amplified a 498 bp amplicon wh...
Seroepidemiological investigation of brucellosis in sheep abortions in Kars, Turkey
Tropical Animal Health and Production, 2009
This study was undertaken to investigate the seroprevalence of brucellosis in unvaccinated sheep from the flocks having previous abortion cases in Kars and around, Turkey and to compare the efficacy of each serological test used. Four hundred serum samples collected from 16 different flocks of sheep having a history of abortions in Kars and its surrounding area in Turkey were examined for the presence of antibodies raised against Brucella using Rose Bengal Plate Test (RBPT), Serum Agglutination Test (SAT), Rivanol Agglutination Test (RAT) and Complement Fixation Test (CFT). All animals were unvaccinated against Brucella. Of the serum samples tested, 147 (%36.7), 142 (%35.5), 139 (%34.75) and 135 (%33.75) were found positive by SAT, RAT, RBPT and CFT, respectively. No statistically significant difference was found between the serological tests used (p > 0.05). It is concluded from this study that brucellosis continues to be an important problem for ovine abortions and poses a risk both for human and other animals in this area. Therefore, adequate intervention measures should be implemented to control and eradicate brucellosis. In addition, if conventional serological tests are used at least two tests, RPBT for screening and CFT for the confirmation of the positive samples are preferable, should be used in parallel for detection of brucellosis effectively.
Abortion is the most obvious manifestation of Brucella infection. In this study, 59 aborted buffalos, 91 aborted cows and 150 aborted women were studied. Diagnosis of Brucella infection in these abortions was based on clinical, serological and bacteriological studies, then determination of Brucella isolates with PCR assay. Serological studies included the use of RB and ELISA tests as screening tests for infection. Results showed that in bovine the RB test shows significant difference between the positive cases in cows and buffalos (P<0.05) while in ELISA test there is no significant difference between these two groups. In aborted women there was significant difference in the RB and ELISA tests between the negative and positive women (P<0.05).From all positive cases by ELISA test, Brucella was isolated from 7 aborted cows, 3 buffalos, and 2 women. Brucella isolates were revealed amplification of a 223-bp fragment with B4 and B5 primers except one strain that isolated from blood culture of women .
Acta Veterinaria Brno, 2008
The prevalence of brucellosis was investigated in cattle, farmers and veterinarians in the Kars district of Turkey between 2004 - 2006. In order to achieve this, a total of 407 serum samples of cattle from 27 herds having history of abortions were examined for Brucella antibodies by RBPT and SAT. In addition, the sera collected from 246 farmers (130 males and 116 females) and 28 veterinarians in the same district were analysed serologically by RBPT, SAT and ELISA. Of the cattle sera analysed, 134 (32.92%) and 141 (34.64%) were determined as positive by RBPT and SAT, respectively. Thirty-two (13%), 35 (14.22%) and 44 (17.88%) of the farmers' sera were found positive for brucellosis by RBPT, SAT and ELISA, respectively. There was no significant difference between sexes for Brucella seropositivity. Of the 28 sera from veterinarians, 13 (46.42%) were positive by the three serological tests. The high prevalence of brucellosis both in cattle and humans suggests that brucellosis is com...
Investigation of bovine brucellosis in the Northeastern Turkey
Tropical Animal Health and Production, 2007
Bovine brucellosis, caused by Brucella abortus, is a significant problem for both public and animal health in Turkey. This study was conducted on the calving seasons between 2001 and 2006. A total of 626 serum samples of cattle obtained from 27 herds with a history of abortions was examined for Brucella antibodies by RBPT, SAT and ELISA. Of the cattle sera analysed, 221 (35,30%) and 206 (32, 92%) and 247 (39,45%) were found to be positive by RBPT , SAT and ELISA, respectively. B. abortus was isolated from 48 (32,21%) of 149 lung samples and stomach contents of the aborted fetuses. Based on the biochemical tests and the agglutination tests with monospecific A and M antisera, only 3 of the isolates were found to be B. abortus biotype 1 and the remaining 45 were biotype 3. This study also revealed that the dominant biotype of B. abortus was biotype 3 in this region. The determination of the agents responsible for bovine brucellosis and serosurvey of this disease are expected to help better understanding of this zoonotic infection in this region and neighbouring countries.
The zoonotic disease brucellosis is a serious public health and livestock industry concern. In the present study, we used bacteriological culture, RBT, and qPCR to determine the prevalence of brucellosis in the serum and milk samples of sheep with a history of abortion. Serum and milk samples were obtained from 100 sheep aged 3-5 years. In order to determine the prevalence of brucellosis, a modified RBT was performed on serum samples, Brucella was isolated from milk by bacteriological culture, and qPCR was applied to detect bacterial DNA in milk. The prevalence of brucellosis using modified RBT, bacteriological culture, and qPCR was 32%, 42%, and 44%, respectively. By considering qPCR as the standard, modified RBT showed a sensitivity of 95%, a specificity of 100%, an accuracy of 98%, a PV+ of 100%, and a PV-of 97%. The sensitivity, specificity, accuracy, PV+, and PV-for bacteriological culture were 77%, 100%, 90%, 100%, and 85%, respectively. The agreement between qPCR and modified RBT was 0.959 (95% CI: 0.896-1), between qPCR and bacteriological culture was 0.792 (95% CI: 0.667-0.897), and between modified RBT and bacteriological culture was 0.831 (95% CI: 0.709-0.38). Based on the results, bacterial isolation from sheep milk is not recommended except in specific cases due to its low sensitivity, as well as its time-consuming and hazardous nature. However, the modified RBT can be used as a routine method because of its cost-effectiveness, higher sensitivity, and higher accuracy compared to bacterial isolation. Moreover, qPCR is recommended as the gold standard test for detecting brucellosis in sheep milk, especially in those with a history of abortion.
2015
Immunochrom atographic test, ELISA. Rapid diagnostic tests are needed to facilitate diagnosis and control of brucellosis. Sero-epidemiology of brucellosis is currently done by employing the Rose Bengal test (RBT). Also the world organization for animal health (OIE) has approved an indirect ELISA for testing serum and milk. The Immuno-Chromatographic brucellosis test (ICT) is a rapid, card-based immunochromatographic test for detection of antibodies directed against B. abortus antigens. To the best of our knowledge, ICT has not been used for the diagnosis of brucellosis in cattle yet. The objective of the study was to evaluate the performance of the ICT brucellosis test for the diagnosis of B. abortus in cattle sera versus RBT and ELISA as a gold standard, Also evaluation of ICT brucellosis test efficacy for detection of antibodies against B. Abortus in milk samples. 94, 90.36 and 84.3 % of sera samples were positive by RBT, ICT and ELISA respectively. The ICT had 94.44 % sensitivity...
Iraqi Journal of Veterinary Medicine, 2016
The aim of this study is to determine the sensitivity of bacterial culture technique in the detection of Brucella abortus in milk samples of aborted cows. Sixty samples of milk were collected from aborted cows during a period which did not exceed two months after the abortion. All of them were positive for rose bengal test. Results showed that Brucella abortus was isolated from 7 out of 60 (11.6%) from the milk of aborted cows, while PCR test showed that 32 out of 60 (53.3%) milk sample contained Brucella abortus. The specificity of culture techniques was 10%, but its sensitivity was only 21.8%. Beside the cautions in dealing with live Brucella abortus (as culture), it is also less sensitive than PCR, though it is better to use PCR technique in the diagnosis of brucellosis in aborted cows milk.