Rose Bengal Plate Agglutination Test Research Articles

Equine brucellosis in Iran: serological, bacteriological and molecular analysis.

Equine brucellosis significantly impacts the health and functionality of horses, leading to complications such as bursitis infection, septic tenosynovitis, septic arthritis, and non-specific lameness resulting from joint infections. In the present study, we used the Rose Bengal plate agglutination test (RBPT), serum agglutination test (SAT), and the 2-mercaptoethanol (2-ME) assays to find equine brucellosis. From June 2018 to September 2022, 876 blood samples were randomly taken from apparently healthy racing horses in certain parts of Iran, such as Kerman, Isfahan, Tehran, Qom, and Kurdistan. DNA extraction was carried out directly on all 63 serum samples identified as seropositive through RBPT. An additional 30 seronegative serum samples were also randomly chosen for study. Bacterial culture was also done on milk, blood, and vaginal swabs taken from seropositive horses.The bacteria that were found in the samples were then put through Bruce-ladder PCR. Our results indicated that 63 (7.1%), 21 (2.3%), and 2 (0.2%) of horses were seropositive using RBPT, SAT, and 2-ME, respectively. Also, none of the 30 DNA-extracted serum samples from seronegative horses tested positive for Brucella DNA, while 44.5% (28/63) of the DNA samples from seropositive horses yielded positive results for Brucella DNA. Out of the seropositive samples, 26 had DNA from Brucella abortus and 2 had DNA from Brucella melitensis. Also, B. melitensis biovar 1 was found in two milk samples from mares in the provinces of Kerman and Isfahan. It was identified using classical biotyping, and molecular assays. It was seen that some of healthy racing horses in some parts of Iran had antibodies against Brucella. The bacteriology and PCR methodologies provide a more comprehensive and reliable means of identifying Brucella spp. infections in horse, especially when the RBPT test came back positive. This underscores the imperative for employing molecular, bacterial, and serological methods in the diagnosis and monitoring of this zoonotic infection. Additionally, this finding suggests that Brucella is being transmitted to equine hosts as a result of its presence in ruminants. The mechanism of transmission may involve interactions between infected ruminants and susceptible equines. This discovery is significant as it underscores the potential cross-species transmission of Brucella and highlights the importance of understanding and managing the spread of the pathogen in both ruminant and equine populations.

Comparison of Fluorescence Polarization Assay with Rose Bengal Plate Agglutination test and Competitive Enzyme-Linked Immunosorbent assay for bovine brucellosis in Tanzania

Brucella are Gram-negative, facultative, intracellular bacterial species with B. abortus, B. melitensis, and B. suis carrying the smooth-lipopolysaccharide antigen. Accurate diagnostic results of brucellosis are needed for its control and eradication, however, they are primarily based on the serological testing of brucellosis in animals. The Rose Bengal Plate Agglutination Test (RBPT) and competitive Enzyme-Linked Immunosorbent Assay (c-ELISA) are the most commonly used tests for making such diagnosis. The use of Fluorescence Polarization Assay (FPA) in Tanzania is still in nascent stage. The purpose of this study was to compare RBPT, c- ELISA, and FPA in the diagnosis of bovine brucellosis. A total of 75 serum samples from cattle that were infected with Brucella in Kagera region were obtained. The FPA showed 90% (68 samples) prevalence, RBPT revealed 93% (70 samples) prevalence, and c-ELISA revealed 81% (61 samples) prevalence of brucellosis in the farm. The RBPT test has shown an inability to distinguish antibodies from cross-reacting organisms compared to the FPA test, while the c- ELISA was unable to pick a positive sample compared to the FPA test. FPA is very quick (5 min per sample), does not require specialized staff, and may be performed under field conditions. Therefore, FPA has a potential to overcome limitations in the detection of bovine brucellosis and can be used as a confirmatory test.

Novel Vertical Flow Immunoassay with Au@PtNPs for Rapid, Ultrasensitive, and On-Site Diagnosis of Human Brucellosis.

Brucellosis is an infectious zoonosis caused by Brucella with clinical symptoms of wavy fever, fatigue, and even invasion of tissues and organs in the whole body, posing a serious threat to public health around the world. Herein, a novel vertical flow immunoassay based on Au@Pt nanoparticles (Au@PtNPs-VFIA) was established for detection of Brucella IgG antibody in clinical serum samples. The testing card of Au@PtNPs-VFIA was manufactured by printing the purified Brucella LPS and goat antimouse IgG on the nitrocellulose membrane as the test-spot or control-spot, respectively. Au@PtNPs labeled with protein G (Au@PtNPs-prG) were concurrently employed as detection probes presenting visible spots and catalysts mimicking catalytic enzymes to catalyze the DAB substrate (H2O2 plus O-phenylenediamine) for deepening color development. The testing procedure of Au@PtNPs-VFIA takes 2-3 min, and the limit of detection (LOD) for Brucella antibody is 0.1 IU/mL, which is faster and more sensitive than that of Au@PtNP-based lateral flow immunoassay (Au@PtNPs-LFIA: 15 min and 1.56 IU/mL, respectively). By comparing with vertical flow immunoassay based on classic Au nanoparticles (AuNPs-VFIA), the Au@PtNPs-VFIA is 32 times or 16 times more sensitive with or without further development of DAB substrate catalysis. Au@PtNPs-VFIA did not react with the serum samples of Gram-negative bacterium infections but only weakly cross-reacted with diagnostic serum of Y. enterocolitica O9 infection. In detection of clinical samples, Au@PtNPs-VFIA was validated for possessing 98.33% sensitivity, 100% specificity, and 99.17% accuracy, which were comparable with or even better than those obtained by the Rose-Bengal plate agglutination test, serological agglutination test, AuNPs-VFIA, and Au@PtNPs-LFIA. Therefore, this newly developed Au@PtNPs-VFIA has potential for rapid, ultrasensitive, and on-site diagnosis of human Brucellosis in clinics.

Evaluation of the Combined Use of Major Outer Membrane Proteins in the Serodiagnosis of Brucellosis.

BackgroundBrucellosis is a zoonotic disease that causes substantial public health problems and endangers the development of animal husbandry in endemic areas. Early diagnosis of infected animals and humans is a crucial step in reducing the incidence of brucellosis. In this study, we designed different combinations of Brucella major outer membrane proteins (omps) including omp10, omp16, omp19, omp25, omp31 and BP26 as antigens and evaluated their efficiency in serodiagnosis for brucellosis. The efficiency assay was conducted using the method of indirect enzyme-linked immunosorbent assay (iELISA) together with a collection of brucellosis-positive sera and healthy sera from multiple species (161 from human, 120 from goat and 144 from cattle). The diagnostic effectiveness of each omp combination was analyzed by receiver operating characteristic (ROC) curve with the software GraphPad Prism version 6.05.ResultsThe omp25/omp31/BP26 combination showed the best efficiency in diagnosis for human brucellosis. The area under the ROC curve (AUC) was 0.995 and, compared with the serum tube agglutination test (SAT) and the Rose Bengal plate agglutination test (RBPT), the positive and negative diagnostic accuracies of iELISA were 94.59% (105/111) and 100.0% (50/50), respectively. Evaluation of the 120 goat and 144 cattle serum samples showed that the best combination for diagnosing both omp31/BP26, the AUC was 0.9262 in goat and 0.9344 in cattle, and compared with those of SAT and RBPT, the positive and negative diagnostic accuracies in goat were 72.73% (48/66) and 100.0% (54/54), respectively. The positive and negative diagnostic accuracies in cattle were 79.79% (75/94) and 100.0% (50/50), respectively. Cross-reaction assays showed that omp25/omp31/BP26 and omp31/BP26 do not cross with other common pathogens.ConclusionThe results indicated that combinations of omps, as protein antigens, can be used to diagnose brucellosis with high accuracy in human, goat and cattle.

Epidemiology and awareness of brucellosis in populations at high risk in Hulunbuir, Inner Mongolia Autonomous Region, 2010–2019

Objective To analyze the surveillance results of brucellosis in high-risk population in Hulunbuir, Inner Mongolia Autonomous Region from 2010 to 2019 and provide evidence for adjusting the brucellosis prevention and control strategy. Methods The stratified sampling surveillance in populations with direct exposures to livestock and its products was conducted in counties of Hulunbuir. Blood samples were taken from the individuals in these counties for preliminary screening by using rose bengal plate agglutination test (RBPT) and for confirmation by using serum agglutination test (SAT). The survey of brucellosis related knowledge awareness was carried out in the above-mentioned populations. The questionnaire included three parts: basic knowledge about brucellosis, risk factors and brucellosis prevention knowledge. Descriptive statistics were used to analyze the infection, incidence and knowledge of brucellosis among high-risk populations. The rate and composition ratio were compared with χ2 test, and P Results From 2010 to 2019, a total of 162 539 blood samples were tested in Hulunbuir, in which 8 556 were positive (5.26%). Over time, the positive rate of blood test showed a downward trend. The difference was significant ( χ2=448.824, P Conclusion The prevention and control strategy for human brucellosis in Hulunbuir has achieved certain progress, but the situation is still serious. Therefore, it is necessary to carry out continuous surveillance for human brucellosis and conduct health education in populations at high risk.

A time-resolved fluorescence lateral flow immunoassay for rapid and quantitative serodiagnosis of Brucella infection in humans

Brucellosis is a worldwide infectious zoonotic disease, posing severe threats to human health and social-economic development. By comparing with time-consuming, low sensitive and non-quantitative conventional serological methods, herein, protein G (prG) coupled with europium nanospheres (EuNPs) (detection probe) and highly purified Brucella lipopolysaccharide (LPS) (capture antigen) were used to develop a novel time-resolved fluorescence lateral flow immunoassay (TF-LFIA) for detecting anti-Brucella IgG antibody in human plasmas. The entire testing took 15 min. With a satisfactory purity, the purified LPS weakly cross-reacted with Y. enterocolitica O9 diagnostic antibody; however, none reacted with sera from patients with other Gram-negative bacterial infections. Following coefficient of determination (R2 = 0.9961), 0.3 IU/mL was reported as the limit of detection (LOD), much lower than those of Serological Agglutination Test (SAT), Rose-Bengal Plate Agglutination Test (RBPT) and colloidal gold LFIA (CG-LFIA). Intra-day and inter-day precisions (CV, coefficient variation) of TF-LFIA varied less than 8% or 12 %, while intra-day and inter-day accuracies were 94–106 % or 93–107 %, respectively. The correlation coefficient (R2) of TF-LFIA measurement to the different concentrations of spiked Brucella antibody was 0.9967, suggesting TF-LFIA had high reliability and reproducibility. TF-LFIA was demonstrated for 100 % specificity, 98.57 % sensitivity and 99.63 % accuracy in detection of Brucella antibody from clinical samples, respectively, significantly higher compared to SAT and RBPT. In conclusion, the established TF-LFIA is a simple, rapid and quantitative immunoassay for early diagnosis or epidemiological surveillance of Brucella infection in humans.

Acute phase biomarkers, oxidants, antioxidants, and trace minerals of mobile sheep flocks naturally infected with brucellosis

This study assayed the acute phase responses of sheep seropositive to Brucella. Sera collected from ewes (n=160) were subjected to serological tests of Brucella, Rose Bengal plate agglutination test (RBPAT), buffer acidified plate agglutination test (BAPAT), and complement fixation test (CFT). Results revealed that CFT was the most predictive test of brucellosis followed by BAPAT then RBPAT. The moderate predictive blood biochemical parameters were zinc and ascorbic acid. Ewes with low CFT titre (chronic) had low fibrinogen, copper, NO, and GPx. Seropositive animals had high blood concentrations of ascorbic acid and zinc.

Pilot study on the camel brucellosis and paratuberculosis in Western India

Brucellosis, a major zoonosis, has attracted attention due to its reemergence. Similarly, paratuberculosis is one of the major health and production problems of animals with zoonotic concerns. Brucellosis and paratuberculosis have been reported from all parts of country with multiple species involvement. The Rose Bengal plate agglutination test (RBPT) and indirect ELISA (iELISA) are most commonly used tests for diagnosing brucellosis. Similarly, plate ELISA and DTH skin tests are popular tests for paratuberculosis. The recently developed penside test is suitable for door step diagnosis of brucellosis. In the present study plate ELISA and penside test were used to investigate brucellosis, and plate ELISA was used to investigate paratuberculosis in camels. Both the plate ELISA and penside tests revealed 4 (11%) animals to be positive for anti-Brucella antibodies. Penside test revealed results at par to plate ELISA test and showed promising results to be adopted for doorstep Brucella sp. testing. Out of 36 camels suspected for paratuberculosis infection, 12 (33.3%) were positive for the paratuberculosis antibodies in plate ELISA. All brucellosis positive animals were negative for paratuberculosis.

Seroprevalence and Molecular Identification of Brucella spp. in Bovines in Pakistan-Investigating Association With Risk Factors Using Machine Learning.

Bovine brucellosis is a global zoonosis of public health importance. It is an endemic disease in many developing countries including Pakistan. This study aimed to estimate the seroprevalence and molecular detection of bovine brucellosis and to assess the association of potential risk factors with test results. A total of 176 milk and 402 serum samples were collected from cattle and buffaloes in three districts of upper Punjab, Pakistan. Milk samples were investigated using milk ring test (MRT), while sera were tested by Rose–Bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (i-ELISA). Real-time PCR was used for detection of Brucella DNA in investigated samples. Anti-Brucella antibodies were detected in 37 (21.02%) bovine milk samples using MRT and in 66 (16.4%) and 71 (17.7%) bovine sera using RBPT and i-ELISA, respectively. Real-time PCR detected Brucella DNA in 31 (7.71%) from a total of 402 bovine sera and identified as Brucella abortus. Seroprevalence and molecular identification of bovine brucellosis varied in some regions in Pakistan. With the use of machine learning, the association of test results with risk factors including age, animal species/type, herd size, history of abortion, pregnancy status, lactation status, and geographical location was analyzed. Machine learning confirmed a real observation that lactation status was found to be the highest significant factor, while abortion, age, and pregnancy came second in terms of significance. To the authors' best knowledge, this is the first time to use machine learning to assess brucellosis in Pakistan; this is a model that can be applied for other developing countries in the future. The development of control strategies for bovine brucellosis through the implementation of uninterrupted surveillance and interactive extension programs in Pakistan is highly recommended.

Frequency of occurrence of Brucellosis in goats in Ludhiana District of Punjab State of India

Background: We investigated the frequency of Brucellosis in goats in Ludhiana district of Punjab state of India because it is of great public health concern yet there is hardly any report in this regard from this region recently. Methods and materials: All the serum samples from goats were subjected to analysis with four common serodiagnostic techniques – RBPT, STAT, MAT and cELISA. Rose Bengal Plate Agglutination Test (RBPT) RBPT was carried out by colored antigen with the method of Morgan et al. Estimation of Antibody Titers by Standard Tube Agglutination Test (STAT). The standard method recommended by OIE was followed. A titer of 1:40 or above was considered positive. Estimation of Antibody Titers by Microtiter Agglutination Test (MAT) MAT was performed as per the standard method as per OIE. The formation of matt signified agglutination while button formation was indicative of a negative reaction. The titer was expressed in the unit system by doubling of the serum titer as International Units (I.U.) giving at least 50 percent agglutination. Competitive Enzyme Linked Immunosorbant Assay (cELISA) All the serum samples were subjected to analysis by competitive ELISA employing a commercial kit Svanovir Brucella Antibody cELISA Kit (Svanova). Results: A total of 191 goats from slaughter houses were analyzed with RBPT, STAT, MAT and ELISA. Out of these, 31 goats were positive for Brucellosis by one or more of these tests. Among positives, 14 were detected by RBPT, 17 by STAT, 21 by MAT, and 21 by ELISA; 10 samples were positive and 160 negative by all. ELISA and MAT detected highest number of samples followed by STAT and RBPT. Frequency of Brucellosis in goats in Ludhiana was found as 5.23% by RBPT and ELISA. The mean titers of anti-Brucella antibodies in goats were 36.66 ± 16.32 by STAT and 49.16 ± 29.47 by MAT, with non-significant difference by ‘t’ test. Conclusion: The present study revealed the frequency of occurrence of Brucellosis in goats in Ludhiana district of Punjab to be 5.23% by RBPT and ELISA. This is alarming considering the zoonotic potential of the disease and high consumption of mutton and milk in this region.

Epidemiological investigations of occurrence of Brucellosis in goats in Ludhiana District of Punjab State of India

Background: Brucellosis in animals is endemic in India, in sheep and goats. I investigated the frequency of Brucellosis in goats in Ludhiana district of Punjab state of India because it is of great public health concern yet there is hardly any report in this regard from this region recently. A total of 191 serum samples of goats from slaughter houses were analyzed with RBPT, STAT, MAT and ELISA. Methods and materials: Rose Bengal Plate Agglutination Test (RBPT). RBPT was carried out by colored antigen with the method of Morgan et al., 10 μl of antigen was mixed with 10 μl of serum and observed for clumps as positive. Estimation of Antibody Titers by Standard Tube Agglutination Test (STAT) The standard method recommended by OIE was followed. 40 IU was taken as positive. Estimation of Antibody Titers by Microtiter Agglutination Test (MAT) MAT was performed as per the standard method. The formation of matt signified positive. The titer was expressed as International Units (I.U.) per milliliter of serum. Competitive Enzyme Linked Immunosorbant Assay (cELISA) All the serum samples were subjected to analysis by competitive ELISA employing a commercial kit Svanovir Brucella Antibody cELISA Kit (Svanova). Statistical analysis The results were analyzed by ANOVA and students ‘t’ test using online software. Sensitivity and specificity were calculated using the standard formulae Results: Out of A total of 191 samples from goats 14 were found to be positive by RBPT, 17 by STAT, 21 each by MAT and ELISA. ELISA and MAT detected the highest number of samples followed by RBPT and STAT, respectively. Frequency of occurrence of the disease in goats was estimated to be 5.23% by RBPT and ELISA.The mean titers of anti-Brucella antibodies in goat sera were 36.66 ± 16.32 by STAT and 49.16 ± 29.47 by MAT, the difference was non-significant by ‘t’ test. Conclusion: The present study revealed the frequency of occurrence of Brucellosis in goats in Ludhiana district, Punjab to be 5.23% by RBPT and ELISA. This is alarming considering the high consumption of mutton and milk in this region.

Clinical features of 9 cases of Brucella endocarditis

Objective: To explore the clinical characteristics of patients with Brucella endocarditis. Methods: The clinical data of 9 patients with Brucella endocarditis admitted to Beijing Ditan Hospital from October 2008 to August 2018 were retrospectively analyzed. Through the electronic medical record system of the hospital. Through assessing the electronic medical record system of the hospital, demographic data, main symptoms, vital signs, blood culture, Rose Bengal Plate Agglutination Test, echocardiography, electrocardiogram, chest imaging and other clinical data of included patients were inquired and recorded. Patients were followed up by telephone for medication, operation and outcome. Results: The 9 patients were all Han nationality, aged from 25 to 66 years, 7 out of 9 patients were male, and they came from Hebei, Shandong, Shanxi, Inner Mongolia and Beijing. Of the 9 patients, 5 were farmers, 2 were self-employed, 1 was a technician, and 1 was unemployed. Of the 9 cases, 8 had a history of close contact with cattle and sheep, and 5 had a history of eating beef and mutton. Rose-Bengal Plate Agglutination Test and blood culture were positive in all 9 patients. Aortic valve was involved in 7 out of 9 patients, mitral and tricuspid valve was involved in 1 patient, respectively, and aortic dissection occurred in 1 patient. Condition of 1 patient rapidly deteriorated after admission and finally died during hospitalization despite antibiotic therapy, the remaining patients received long-term antibiotic treatment. A total of 7 patients who underwent valve replacement were followed up. One patient died of cerebral hemorrhage 6 months after operation, and the remaining 6 patients recovered well after valve replacement. Heart failure occurred in all 9 patients, and pericardial effusion occurred in 8 patients. Electrocardiogram showed low voltage of the QRS complex in the limb in 3 cases and poor R-wave progression in V(1)-V(3) lead in 2 cases, and sinus tachycardia in 2 cases. One patient developed non-specific ST-T abnormalities. All patients had fever, 7 patients complained of weakness, and 6 patients complained of palpitations. Among the 9 patients, 7 cases had anemia, 7 patients had pneumonia, 6 had bilateral pleural effusion, 4 had thrombocytopenia. Creatinine was above normal in 4 patients, urine protein was positive in 3 patients, Delta Bilirubin was higher in 3 cases. Conclusions: Patients with Brucella endocarditis often suffer from heart failure and have severe complications. Adequate antibiotic therapy in combination with valve replacement is effective for the treatment of patients with Brucella endocarditis.

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan.

Bovine brucellosis remains a persistent infection in ruminants in Pakistan. A total of 828 (409 buffaloes and 419 cattle) sera were collected from 11 institutional-owned livestock farms in Punjab, Pakistan. The samples were tested by rose bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). The seroprevalence along with 95% confidence interval (CI) was determined. Univariable and multivariable analysis of the epidemiological background data was conducted and odds ratio (OR) was calculated to understand any association between the risk factors and the seroprevalence. An overall seroprevalence of 3.9% (Positive/Tested = 32/828) and 3.3% (27/828) was detected by RBPT and iELISA, respectively. The seroprevalence of 5.6% (CI 3.6–8.3) and 4.7%, (CI 2.8–7.2) and the odds ratio of 2.63 (CI 1.20–5.77) and 2.50 (CI 1.08–5.78) for testing positive by RBPT and iELISA, respectively were significantly higher (p < 0.05) in buffaloes than in cattle. Breed, sex, history of abortion and retention of fetal membranes (RFM) in the animals were not found statistically significantly associated with the infection. RBPT and iELISA based results agreed almost perfect (k = 0.877). In total, Brucella abortus-DNA (9/27) was amplified from seropositive samples by real-time polymerase chain reaction. This study identified for the first time the etiological agents of brucellosis at a molecular level at institutional-owned livestock farms in Pakistan.

Seroprevalence of equine glanders in horses in the central and eastern parts of Mongolia.

Glanders is a contagious and fatal equine disease caused by the gram-negative bacterium Burkholderia mallei. B. mallei is prevalent among horse populations in Asia, the Middle East, and South America. More than four million horses have been registered in Mongolia in 2020. However, the recent prevalence of glanders has not been well investigated. In this study, we aimed to investigate the seropositivity of B. mallei in horse populations in Mongolia using the complement fixation test (CFT) and Rose Bengal plate agglutination test (RBT). We randomly collected blood samples from horses in central and eastern Mongolia between 2018 and 2019. Of 337 horses, 26 (7.7%) and 28 (8.3%) were seropositive using RBT and CFT, respectively. Interestingly, seropositivity in horses resulting from crossbreeding of Mongolian native horses with thoroughbred horses was higher than that in Mongolian native horses. Our observations suggest that equine glanders are still endemic to Mongolia.

A cross-sectional seroepidemiological study of camel (Camelus dromedarius) brucellosis and associated risk factors in the Sultanate of Oman.

Brucellosis is a globally distributed and economically devastating zoonotic disease of multiple species, including camels. Human and livestock brucellosis is prevalent in Oman, especially in southern Dhofar governorates of Oman, where camels share habitat and have close contact with other susceptible species. We conducted a randomized cross-sectional sero-epidemiological study to investigate the seroprevalence of brucellosis in camels of Oman. The sera from 2,250 camels from 552 geographically marked farms were screened through Rose Bengal plate agglutination test and later confirmed by the competitive ELISA (COMPLIZA, VLA, UK). In total, nine [0.4%, confidence interval (CI) 0.4, 0.8] camels from eight (1.5%, CI 0.6, 2.8) herds were tested positive for brucellosis. The highest prevalence was recorded in Dhofar (3.7%, CI 1.4, 7.9) and the lowest in Sharqiyah (1.3%, CI 0.0, 7.2) governorate (p = 0.052). All seropositive camels were of local breed and females. Seroprevalence was higher (0.5%, CI 0.2, 1.0) in adults (>4 yr of age) as compared with young (≤4 yr of age) camels (0.2%, CI 0.0, 0.8). The results of binary logistic regression indicated that camel herds located in south (Dhofar) [odds ratio (OR) 6.39, CI 1.01, 40.67], practice of open replacement system (OR 14.49, CI 1.83, 114.51) and with history of abortions (OR 8.01, CI 1.34, 47.77) were more likely to test positive for brucellosis. We conclude that brucellosis is endemic at a low level in camels of Oman and a control strategy based upon test-and-slaughter/inclusion of camels in the current vaccination program after carefully planned vaccine evaluation studies could be considered to control it.

"Confounding of Brucellosis and Blue Tongue in Trans Humane Sheep Flock of Tamilnadu and other Herd Health Impact"

This present study was carried out to assess the prevalence of brucellosis and blue tongue in a trans humane sheep flock of Tamil Nadu, India. This Sheep flock had a history of inconsistent abortion, repeat breeder, poor fertility rate and higher prevalence of still birth. Serum samples were collected from sheep by random sampling. Serum samples were subjected to Rose Bengal Plate agglutination test (RBT) and ELISA. The risk factors like pregnancy, abortion, age and sex were correlated to the Brucella seropositivity. This study also assessed for the presence of Bluetongue in aborted sheep. It was found that ELISA could be the choice of test for testing of Brucellosis (with the percentage of 57.14). Clinically healthy rams were found to be with brucellosis seropositivity and posed infertility to ewes. It was observed that in trans humane flocks. Brucellosis and Blue tongue has a confounding phenomenon for ovine abortions.

Comparative Seroepidemiological Study of Brucellosis in Sheep under Smallholder Farming and Governmental Breeding Ranches of Central and North East Ethiopia.

This cross-sectional study was carried out to investigate the status of brucellosis in sheep management under extensive smallholder farming and intensively in governmental breeding ranches in six districts selected from three administrative zones. Using multistage sampling, serum samples of 2409 sheep from 274 flocks were collected and tested using the Rose Bengal Plate Agglutination Test (RBPT) and positive sera were confirmed using a Complement Fixation Test (CFT). Of all animals tested, 4.98% were RBPT positive, and after confirmation with CFT, the overall animal-level prevalence was found to be 4.89% (CI: 3.24-6.9%). Of the flocks sampled, 61 (22.3%, CI: 18.03-29.17%) had at least one animal positive to both tests. Significantly higher (P < 0.001) individual animal seroprevalence of 5.87% (CI: 3.83-7.31%) was found in sheep under smallholder production than in breeding ranches (1.75%, 95% CI: 1.57-3.05%). However, flock level seroprevalence in breeding ranches was found to be 100% (8/8), while in the smallholder production it was 19.92% (CI: 16.4-25.81%). Significantly highest animal-level seroprevalence of 9.55% (CI: 7.91-12.4%) was observed in north Wollo zone's smallholder farms. From the three studied breeding ranches, highest seroprevalence of 3.57% (CI: 2.84%-5.18%) was found in Sheno Agricultural Research Centre. Significantly higher seroprevalence (P < 0.01) was found in aborted sheep and with history of retained fetal membrane in both production systems. All the sheep flocks in the studied breeding ranches were found to be seropositive; hence, this study suggests strict control measures of ovine brucellosis in the breeding reaches, since they could be a source of infection for the smallholder farms.

The Surgical Treatment of Lumbar Brucellar Spondylitis Byposterior Approach

Objective: This study investigate the effect of surgical treatment of lumbar spondylitis, which used posterior approach to remove the lesion, graft the bone between transverse process and used nail rod system to make Internal fixation. Methods: From February 2011 to April 2017, 62 cases of lumbar brucellar spondylitis were analyzed retrospectively in the No. 1 Affiliated Hospital of Hebei North University in Zhangjiakou City, Hebei Province, China. Including 37 male and 25 female patients aged 24 to 61; average age was (42.8 ± 1.5). With an average duration of 4-28 months, average month was (6.6 ± 1.8). All the patients have the history of contact the source of animal, impurity diet history or other pathogen exposure history. The result of X-ray, CT, MRI, c-reactive protein and blood sedimentation, rose bengal plate agglutination test(RBPT) and standard tube agglutination test(SAT) check were positive. The outcome also conforms to National Health and Family Planning Commission of the People`s Republic of China WS 269-2007 brucellosis diagnostic criteria and principles of treatment. Preoperative, doxycycline + rifampicin + sulfamethoxazole were used for 56 days. Next, Surgical treatment was performed by using posterior approach to remove the lesion, graft the bone between transverse process and used nail rod system to make Internal fixation. Post-operative, the anti-bulimia drugs was used again until the C-reactive protein, blood pressure, RBPT test and the SAT test were negative. Furthermore, other 2 weeks medication should be used necessary. During drugs oral therapy time, the monitoring indexes of related infections were dynamically rechecked to monitor liver and kidney function. Visual analogue scale (VAS) was used to assess the degree of lower limb pain and lower back pain and Oswestry disability index (ODI) was used to assess the degree of recovery of waist function, Frankel spinal cord injury classification method was used to determine the recovery of spinal cord function. Results: The operation time was 165-235 min, averaging (175.3 ± 22.1) min. Intraoperative bleeding was 260 ~ 510ml, averaging (338.3 ± 75.9) ml. The length of hospitalization was 8 ~ 14days, on average (11.99 ± 3.25) days. No intraoperative vascular injury or spinal cord injury. The incisions were healed well. The follow-up postoperative was more than 12 months, averaging month is 18.2 ± 4.5. The infection symptoms disappeared. After 3 months, the blood sedimentation and other infection indexes were returned to normal. The result of X-ray examination showed that bone healing was obtained in transverse process between 3 ~ 12 months, and the fusion time of bone graft was 7 ~ 11 months, average month is (8.4 ± 1.2). After 3 months, 6 months and 12 months follow-up, VAS score points was (3.05 ± 0.65), (1.88 ± 0.55) and (0.31 ± 0.42) respectively. Compared with preoperative points (6.67 ± 1.85), which is significantly lower. The results was statistically significant (P = 0.012); the classification of Frankel spinal cord injury was significantly better than before (P=0.022). The ODI functional disorder index was (21.15 ± 5.35), (12.26 ± 4.55) and (5.31 ± 3.12), lower than preoperative significantly (37.87 ± 4.85), and the results were statistically significant (P=0.031). Conclusion: We could remove the lesion, graft the bone between transverse process and use nail rod system to make internal fixation by posterior approach, thoroughly remove the lesion which was granulation tissue or abscess of lumbar spine or around the spinal canal. For the vertebral canal and nerve root canal, a complete decompression can be created. After using nail rod system to make internal fixation, the lumbar spine could have an immediately stabilized. This method could recover the normal physiological protuberance of lumbar spine and prevent the instability occur in lumbar spine. For lumbar Brucella spondylitis, combine with long time oral therapy and posterior approach operation was an effective treatment method.

The Surgical Treatment of Lumbar Brucellar Spondylitis By posterior Approach

Objective: This study investigates the effect of surgical treatment of lumbar spondylitis, which used posterior approach to remove the lesion, graft the bone between transverse process and used nail rod system to make Internal fixation. Methods: From February 2011 to April 2017, 62 cases of lumbar brucellar spondylitis were analyzed retrospectively in the No. 1 Affiliated Hospital of Hebei North University in Zhangjiakou City, Hebei Province, China. Including 37 male and 25 female patients aged 24 to 61, average age was (42.8 ± 1.5). With an average duration of 4-28 months, average month was (6.6 ± 1.8). All the patients have the history of contact the source of animal, impurity diet history or other pathogen exposure history. The result of X-ray, CT, MRI, c-reactive protein and blood sedimentation rose Bengal plate agglutination test (RBPT) and standard tube agglutination test (SAT) check were positive. The outcome also conforms to National Health and Family Planning Commission of the People`s Republic of China WS 269-2007 brucellosis diagnostic criteria and principles of treatment. Preoperative, doxycycline + rifampicin + sulfamethoxazole was used for 56 days. Next, Surgical treatment was performed by using posterior approach to remove the lesion, graft the bone between transverse process and used nail rod system to make Internal fixation. Post-operative, the anti-bulimia drugs was used again until the C-reactive protein, blood pressure, RBPT test and the SAT test were negative. Furthermore, other 2 weeks medication should be used necessary. During drugs oral therapy time, the monitoring indexes of related infections were dynamically rechecked to monitor liver and kidney function. Visual analogue scale (VAS) was used to assess the degree of lower limb pain and lower back pain and Oswestry disability index (ODI) was used to assess the degree of recovery of waist function, Frankel spinal cord injury classification method was used to determine the recovery of spinal cord function. Results: The operation time was 165-235 min, averaging (175.3 ± 22.1) min. Intraoperative bleeding was 260 ~ 510 ml, averaging (338.3 ± 75.9) ml. The length of hospitalization was 8 ~ 14 days, on average (11.99 ± 3.25) days. No intraoperative vascular injury or spinal cord injury. The incisions were healed well. The followup postoperative was more than 12 months; averaging month is 18.2 ± 4.5. The infection symptoms disappeared. After 3 months, the blood sedimentation and other infection indexes were returned to normal. The result of X-ray examination showed that bone healing was obtained in transverse process between 3 ~ 12 months, and the fusion time of bone graft was 7 ~ 11 months, average month is (8.4 ± 1.2). After 3 months, 6 months and 12 months follow-up, VAS score points were (3.05 ± 0.65), (1.88 ± 0.55) and (0.31 ± 0.42) respectively. Compared with preoperative points (6.67 ± 1.85), which were significantly lower. The results was statistically significant (P=0.012); the classification of Frankel spinal cord injury was significantly better than before (P=0.022). The ODI functional disorder index was (21.15 ± 5.35), (12.26 ± 4.55) and (5.31 ± 3.12), lower than preoperative significantly (37.87 ± 4.85), and the results were statistically significant (P=0.031). Conclusion: We could remove the lesion, graft the bone between transverse process and use nail rod system to make internal fixation by posterior approach, thoroughly remove the lesion which was granulation tissue or abscess of lumbar spine or around the spinal canal. For the vertebral canal and nerve root canal, a complete decompression can be created. After using nail rod system to make internal fixation, the lumbar spine could have an immediately stabilized. This method could recover the normal physiological protuberance of lumbar spine and prevent the instability occur in lumbar spine. For lumbar Brucella spondylitis, combine with long time oral therapy and posterior approach operation was an effective treatment method.

Exposure of harbour seals Phoca vitulina to Brucella in declining populations across Scotland.

Since 2000 there has been a major decline in the abundance of Scottish harbour seals Phoca vitulina. The causes of the decline remain uncertain. The aim of this study was to establish the extent to which the seals in the regions of greatest decline have been exposed to Brucella, a bacterial pathogen that causes reproductive failure in terrestrial mammalian hosts. Tissues from dead seals collected between 1992 and 2013 were cultured for Brucella (n = 150). Serum samples collected from live capture-released seals (n = 343) between 1997 and 2012 were tested for Brucella antibodies using the Rose Bengal plate agglutination test (RBT) and a competitive enzyme-linked immunosorbent assay (cELISA). In total, 16% of seals cultured had Brucella isolated from one or more tissues, but there were no pathological signs of infection. The cELISA results were more sensitive than the RBT results, showing that overall 25.4% of seals were seropositive, with the highest seroprevalence in juveniles. As there was no evidence of either a higher seroprevalence or higher circulating antibody levels in seropositive animals in the areas with the greatest declines, it was concluded that Brucella infection is likely not a major contributing factor to recent declines. However, the consistently high proportion of seals exposed to Brucella indicates possible endemicity in these populations, likely due to B. pinnipedialis, which has demonstrated a preference for pinniped hosts. Importantly, given the close proximity between seals, humans and livestock in many areas, there is the potential for cross-species infections.