Serum Tube Agglutination Test Research Articles

Seroprevalence and Molecular Characterization of Brucella abortus from the Himalayan Marmot in Qinghai, China.

Brucellosis is a serious public health issue in Qinghai (QH), China. Surveying the seroprevalence and isolation of B. abortus strains from marmots is key to understanding the role of wildlife in the maintenance and spread of brucellosis. In this study, a set of methods, including a serology survey, bacteriology, antibiotic susceptibility, molecular genotyping (MLST and MLVA), and genome sequencing, were employed to characterize the two B. abortus strains. The seroprevalence of brucellosis in marmots was 7.0% (80/1146) by serum tube agglutination test (SAT); one Brucella strain was recovered from these positive samples, and another Brucella strain from a human. Two strains were identified as B. abortus bv. 1 and were susceptible to all eight drugs examined. The distribution patterns of the accessory genes, virulence associated genes, and resistance genes of the two strains were consistent, and there was excellent collinearity between the two strains on chromosome I, but they had significant SVs in chromosome II, including inversions and translocations. MLST genotyping identified two B. abortus strains as ST2, and MLVA-16 analysis showed that the two strains clustered with strains from northern China. WGS-SNP phylogenetic analysis showed that the strains were genetically homogeneous with strains from the northern region, implying that strains from a common lineage were spread continuously in different regions and hosts. Seroprevalence and molecular clues demonstrated frequent direct or indirect contact between sheep/goats, cattle, and marmots, implying that wildlife plays a vital role in the maintenance and spread of B. abortus in the Qinghai-Tibet Plateau.

A follow-up study of 100 patients with acute brucellosis for its prognosis and prevention.

To prevent chronic brucellosis, this study analysed the changes in patient antibody titers, and the trajectories of biochemical indicators at different stages of brucellosis, identified relevant biomarkers, and explored risk factors affecting the prognosis of brucellosis patients. A prospective cohort study was conducted to follow 100 patients with acute brucellosis. Laboratory serological test results [taken with a serum (tube) agglutination test (SAT)] and biochemical parameters (liver function, renal function, and hematological system) were measured repeatedly at four-time points: 0 weeks-baseline survey, 6 weeks after the first treatment, 12 weeks after the second treatment, and 3 months after the third treatment. The changes in the antibody titres and biochemical parameters at each time point were analysed for trend changes. One hundred patients with acute brucellosis were enrolled in this follow-up study, with 100% retention in follow-up. By the third follow-up, 21 patients had turned subacute and 11 had turned chronic. One-way repeated measures analysis of variance results showed statistically significant differences (p < 0.01) across the time points for the following five indicators: alanine aminotransferase, aspartate aminotransferase, total bilirubin, serum creatinine (SCr) and platelet count. The clinical symptoms of patients in the acute stage were mainly joint pain, fatigue, and fever, while those in the chronic stage complained primarily of joint pain and fatigue. The results of multivariate logistic analysis showed that joint pain [odds ratio (OR) = 3.652, 95% confidence interval (CI) =1.379-9.672], monoarticular pain (OR = 6.356, 95% CI = 4.660-8.669), elevated SCr (OR = 15.804, 95% CI = 1.644-151.966) and elevated haemoglobin (Hb) (OR = 1.219, 95% CI = 1.065-1.736) were risk factors for poor prognosis (not cured or chronic) in patients with brucellosis. The trajectory of changes in patient SAT posirates and antibody titers can be used to distinguish patients with chronic brucellosis. The brucellosis is preventable and treatable, and the standard treatment can be effective in reducing the clinical symptoms of affected patients. If patients are not treated in a timely manner, joint pain, monoarticular pain, and elevated SCr are risk factors for patients who are not cured. Therefore, the treatment cycle for these patients should be extended.

The role of the serum tube agglutination test in the monitoring of human brucellosis: evaluation of post-treatment SAT titers.

SUMMARYOBJECTIVE:Positive results of the serum tube agglutination test that persist after treatment may be interpreted by clinicians as treatment failures. Therefore, our study examined the value of serum tube agglutination test in demonstrating treatment success.METHODS:In this retrospective study conducted at a single center, the pre- and post-treatment serum tube agglutination test titers of patients diagnosed with brucellosis were compared.RESULTS:The end-of-treatment serum tube agglutination test titer was negative in 24 (18%) of 139 patients diagnosed with brucellosis. The most common complaints of the patients were fever (78.4%), chills (88.5%), sweating (84.9%), anorexia (79.1%), and arthralgia (63.3%). The rate of positive blood culture before the treatment was 68.3%. The absence of fever (p=0.005) and arthralgia (p=0.024) and the pretreatment serum tube agglutination test titer of <1/160 (p=0.014) were significant markers of serological cure.CONCLUSION:Although serum tube agglutination test is an effective and very successful test in the diagnosis of brucellosis, our study shows that serum tube agglutination test is not useful in demonstrating the treatment success of human brucellosis in the early post-treatment period.

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.

English

This study was conducted to diagnose the seroprevalence of Listeriosis, Brucellosis, and Toxoplasmosis in camels from Kirkuk province in Iraq. Seventy-six serum samples were randomly collected and analyzed from camels from April to September 2018. Three serological diagnostic methods were used; Osebold agglutination test (OAT), Serum tube agglutination test (SAT) and Sabin- Feldman dye test (SFDT). From all serum samples that were collected, 42(55.26%) showed infections with Listeria, Brucella, and Toxoplasma as following 15(19.7%), 7(9.2%), and 20(26.3%) respectively. Additionally, the study revealed a highly significant relationship between aborted camels and their infection with Brucellosis and Toxoplasmosis (58.3%). In contrast, infections with these pathogens have no significant relationship with the sex and age of the camels. Risk factors of abortion were related to the high prevalence of Brucellosis and Toxoplasmosis in camels' herds. Furthermore, this study found out that the camels in Kirkuk city might be the source of these infectious agents and disseminated the diseases to the human through their contaminated dairy products and meat.

A Retrospective Survey of the Abortion Outbreak Event Caused by Brucellosis at a Blue Fox Breeding Farm in Heilongjiang Province, China.

Brucellosis is a common zoonosis in China, resulting in abortion in animals. Outbreaks of abortion in blue foxes caused by Brucella infection have rarely been reported. In the present study, 3–5 mL blood samples collected from the femoral veins of 10 abortuses of blue foxes were assessed by RBPT (Rose Bengal plate test) and SAT (serum tube agglutination test) to preliminarily investigate the source of infection for the clustering of abortion events at a blue fox farm in Heilongjiang Province. Screening experiments showed that all 10 blood samples were positive in the RBPT, while only eight blood samples out of the 10 were positive in the SAT. Subsequently, 10 tissue samples (spleen, lungs, stomach contents, and afterbirth) from the same 10 foxes were assessed using AMOS (acronym for B. abortus, melitensis, ovis, and suis)-PCR (polymerase chain reaction), and sequencing analysis was performed on amplification products to verify the results of the serology survey. Results showed a spectral band of ~731 bp in these samples. BLAST showed sequences of AMOS-PCR products in this study to be 100% similar (E = 0.0) to sequences in B. melitensis strain from GenBank. These data preliminarily indicated that the blue fox's outbreak of abortion events was caused by brucellosis via the B. melitensis strain. Then 726 serum samples were tested by RBPT and SAT to determine the prevalence of brucellosis on the farm. A comprehensive epidemiological and reproductive status survey of the infected blue fox population was performed. The seropositive rate was found to be 67.90% (493/726) by RBPT and 41.32% (300/726) by SAT. The technicians had stopped feeding the foxes with chicken carcasses and instead fed them raw ground sheep organs (lungs, tracheae, placentae, and dead sheep fetuses) infected by B. meliteneis strains, and that this change in diet caused the outbreak of abortion events. The high abortion rate (55%) and low cub survival rate (65%) were the most distinctive features of the outbreak; these factors led to severe economic losses. Feeding cooked sheep/goat offal and strict breeding management is necessary for disease prevention.

Zoonotic brucellosis: Seroprevalence and different serological tests comparison in ovine and caprine population in district Quetta, Balochistan

The present study was design to determine the seroprevalence of zoonotic brucellosis and different serological tests comparison in ovine and caprine population in district Quetta, Balochistan. A total of 500 blood samples, comprising of 250 each from sheep and goat were randomly collected from out skirts of District Quetta, Balochistan. Out of the 250 blood samples 125 were collected from each males and females. The serum samples were tested for the presence of anti-Brucella antibodies by Rose Bengal Plate Test (RBPT), Serum Plate Agglutination Test (SPAT) and Serum Tube Agglutination Test (STAT). The overall prevalence of brucellosis in sheep was recorded as 16.4%, 13.2% and 10.8% by RBPT, SPAT and STAT, respectively and in goat was found to be 11.6%, 8.8% and 6.8% by RBPT, SPAT and STAT, respectively. The sex-wise prevalence of brucellosis in Ram (male sheep) was recorded as 12.8%, 8.8% and 7.2% by RBPT, SPAT and STAT, respectively; while in Ewe (female sheep) it was 20.0%, 17.6% and 14.4% by RBPT, SPAT and STAT respectively. In Buck (male goat), the seroprevalence of brucellosis was recorded 8.0%, 5.6% and 3.2%, whereas in Dew (female goat) it was 15.2%, 12.0% and 10.4% by RBPT, SPAT and STAT, respectively. The prevalence of brucellosis was relatively higher in ovine (sheep) as compared to caprine (goat), irrespective of techniques used. It was also concluded that brucellosis was higher in females than in males in both sheep and goat. Among all the serological tests applied in the present study, RBPT was found to be more sensitive and showed higher prevalence of brucellosis in sheep as well as in goat population.

Expression of heat shock proteins (HSPs) in caprine bone marrow- derived mesenchymal stem cells

The purpose of the study was to determine seropositivity for brucellosis among sheep and goats having history of abortion and various reproductive disorders. A total of 339 serum samples (136 sheep and 203 goats) were tested for Brucella antibody by Rose Bengal Plate Test (RBPT), Serum Tube Agglutination Test (STAT) and Sandwich Enzyme Linked Immunosorbent Assay (s-ELlSA). The prevalence of brucellosis by RBPT, STAT and s-ELISA was4.41%, 5.88% and 8.08% in sheep and 17.24 %, 8.37% and 12.81% in goats, respectively. Among the three serological tests employed for detection of Brucella antibodies, s-ELISA yielded maximum (13.57%) positive results followed by STAT (10.03%) and RBPT (6.78%). Thus s-ELISA is proved to be the best for screening of animals for brucellosis.

Predictors of Blood Culture Positivity in Pediatric Brucellosis.

To investigate the predictors of blood culture positivity in children with brucellosis. Descriptive study. Department of Pediatric Infectious Diseases in Erzurum Regional Training and Research Hospital, Turkey, from January to December 2015. Eighty-six children under 16 years of age, with brucellosis, were retrospectively evaluated. Compatible clinical findings plus presence of positivity at titers of >1:160 in serum and/or Coombs agglutination tests in a single serum sample and/or a minimum fourfold increase within a 2-3 week interval were diagnostic for brucellosis. Only patients with blood cultures were included. Patients' demographical, clinical, and laboratory risk factors, such as age, gender, presence and duration of symptoms, and laboratory characteristics were analysed. Brucella spp. grew in blood cultures of 24 (27.9%) patients. Children with blood culture positivity had shorter symptom duration than those with negative blood cultures (p=0.03). Absence of personal and household histories of brucellosis (p=0.02 and p=0.04, respectively), lower hemoglobin, iron, and vitamin D (p<0.001, p=0.006, and p=0.006, respectively), and higher leukocyte, CRP, and ferritin (p<0.001, p=0.001, and p<0.001, respectively) levels were associated with isolation of Brucella spp. in blood culture. Children with positive blood cultures had higher serum tube and Coombs agglutination test results (p=0.001 and p<0.001). ROC analysis showed that ferritin at a cut-off level of 122 ng/mL (CI 95% 0.86-0.97, p<0.001) and Brucella Coombs agglutination test at a cut-off level of 1/480 (CI 95% 0.84-0.96, p<0.001) were the most sensitive and specific predictors of bacteremia. Serum hemoglobin, iron, ferritin, vitamin D, and C-reactive protein levels, Brucellar tube and Coombs agglutination tests, and leukocyte count could help to predict definitive diagnosis in pediatric brucellosis when molecular techniques are not feasible, such as in source-limited countries.

BRUCELLOSIS

Background: Human brucellosis is a zoonotic infectious disease transmitted from domesticated animals to humans. It remains a major public health problem in Saudi Arabia where 7% of the population still maintains a nomadic life style with domestication of animals. Objectives: This study aims to discover the prevalence of brucellosis and Brucella among patients attending a district government hospital in Majmaah, kingdom of Saudi Arabia. Period: 2 years (from 1st February 2016 to 31st January 2018). Design: Cross sectional descriptive. Settings: King Khalid Hospital, Al Majmaah, Saudi Arabia. Materials (Patients) &amp; Methods: Total 1098 serum samples from the patients with clinical suspicion of brucellosis were screened with rapid slide agglutination test (crescent diagnostics Jeddah) for Brucella antibodies and later confirmed by Serum Tube Agglutination Test. Main Outcome Measures: The result of our study showed that the prevalence of brucellosis among patients attending our study is 9.1%. Results: 100 individuals were found to be seropositive with titers ≥1:160. The result of our study showed that the prevalence of brucellosis among patients attending our study is 9.1%The Majority of the patients presented to the medical OPD and orthopedic unit of the hospital. The prevalence of disease in males was found to be higher as compared to females in the current studied group. The age group 21- 40 years, was found to be far more susceptible to this infection. Majority presented with fever and musculoskeletal complaints. The total seroprevalence rate calculated for the patients attending our hospital is 26.50%. Conclusion: Frequent serological surveillance should be carried out in areas that are endemic. Screening of risk groups, imported animals and household members of active brucellosis must be undertaken. This is a crucial epidemiological move allowing for timely diagnosis and control of disease. Limitations: Our prevalence rate represents a specific segment of the population (i.e., those attending the hospital) and not thegeneral population.

The effects of the conjunctival Brucella vaccine on some biochemical parameters in sheep.

The genus Brucella causes significant economic losses due to infertility, abortion, stillbirth or weak calves, and neonatal mortality in livestock. Brucellosis is still a zoonosis of public health importance worldwide. In the past, vaccination was administered subcutaneously and nowadays, the conjunctival vaccine is administered. There is no definite information about the changes of the biochemical parameters and antibody response after conjunctival vaccination. In this study, the investigation of the changes in the levels of some biochemical parameters due to the conjunctival vaccination for brucellosis was aimed. Thirty sheep were used as an animal material. The vaccine was done single dose against Brucella melitensis and the blood was drawled from Vena jugularis during 4months. Antibody levels were determined by serum tube agglutination test. Alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), glucose, total protein, and albumin levels were measured using commercial autoanalyzer in sera. The antibody titers (p < 0.001) increased significantly at first month compared to the pre-vaccination, but at the second month began to fall. There was no statistically significant changes in glucose, AST, ALT activity after vaccination (p > 0.005). The significant amount of total protein and ALP decreased after vaccination (p < 0.005). LDH levels and total protein levels were significantly increased (p < 0.005). In conclusion, conjunctival vaccine was considered to be used as a safe to protect the sheep from brucellosis and the results of the study may be used to improve the efficiency of brucellosis eradication programs within livestock management.

A case report of spontaneous abortion caused by Brucella melitensis biovar 3

BackgroundBrucellosis is a worldwide zoonotic disease caused by Brucella spp. Brucella invades the body through the skin mucosa, digestive tract, and respiratory tract. However, only a few studies on human spontaneous abortion attributable to Brucella have been reported. In this work, the patient living in Shanxi Province in China who had suffered a spontaneous abortion was underwent pathogen detection and Brucella melitensis biovar 3 was identified.Case presentationThe patient in this study was 22 years old. On July 16, 2015, she was admitted to Shanxi Grand Hospital, Shanxi Province, China because of one day of vaginal bleeding and three days of abdominal distension accompanied by fever after five months of amenorrhea. A serum tube agglutination test for brucellosis and blood culture were positive. At the time of discharge, she was prescribed oral doxycycline (100 mg/dose, twice a day) and rifampicin (600 mg/dose, once daily) for 6 weeks as recommended by the World Health Organization (WHO). No recurrence was observed during the six months of follow-up after the cessation of antibiotic treatment.ConclusionsThis is the first reported case of miscarriage resulting from Brucella melitensis biovar 3 isolated from a pregnant woman who was infected through unpasteurized milk in China. Brucellosis infection was overlooked in the Maternity Hospital because of physician unawareness. Early recognition and prompt treatment of brucellosis infection are crucial for a successful outcome in pregnancy.

The Senile Brucellosis Spondylitis Clinical Diagnosis and Treatment on as Evaluation Were Retrospectively Analyzed

Objective: To explore senile brucellosis spondylitis clinical features and diagnostic criteria, in order to improve the diagnosis rate and evaluate the clinical effects of treatment strategies. Methods: From January 2002 to August 2015, 62 patients with Brucella-associated spondylitis were treated with comprehensive diagnosis based on epidemiological history, clinical manifestations, imaging findings, laboratory tests, and local biopsy. The positive rate of red plate agglutination test (RBPT) was 45.1%, the positive rate of serum tube agglutination test (SAT) was 51.6%, and the positive rate of brucellosis anti-human immunoglobulin test (Coomb’s) was 100%. All patients underwent X-ray, CT and MRI examinations. The positive rate of blood culture in pathogenic examination was 16.1%, and the positive rate of bone marrow culture was 38.5%. The positive rate of inflammatory granuloma or abscess in the lesion sampling culture was 73.6%. All patients underwent standardized drug therapy and hyperbaric oxygen adjuvant therapy. Among them, 38 patients with neurological impairment were treated with one-stage debridement and posterior pedicle screw fixation on the basis of drug therapy. The paraspinal abscess and the diseased in-tervertebral space tissue were taken for pathological examination during the operation. One week, two weeks, one month, three months, six months and 12 months after treatment, the patients were evaluated and followed up at the monitoring sites. The evaluation indicators were pain scores, activity of daily living (ADL), imaging findings, and laboratory tests. The SPSS15.0 statistical package was used for analysis. Results: All patients were confirmed by the above-mentioned comprehensive examination after admission. 24 patients (38.71%) were treated with standard drug therapy (group A), no adverse drug reactions and abnormal liver and kidney function; the remaining 38 patients (61.29%) were combined with varying degrees of neurological impairment, and surgical treatment was performed after 2 to 4 weeks of drug therapy without improvement (group B). The pain scores showed that there was a statistically significant difference between the two groups at the same time point (P < 0.05). The B group was superior to the A group in both treatment time and pain relief. The blind test evaluation of imaging observation showed that in group B, except for the slow repair of the lesion area, the spine was stable without abscess, sequestrum and inflammatory granulomas, and the score area was stable at 4 - 5 points; in group A, after 3 months of treatment, infections in the vertebral body and intervertebral space were not significantly absorbed, and the infection gradually absorbed after 6 months. Laboratory tests showed that over time, the SAT titer, CRP turnover rate, and RBPT negative rate in group B were significantly better than those in group A, indicating that treatment in group B could reduce SAT titer, CRP, and negative RBPT earlier. The clinical curative effect evaluation of 12 months showed that the clinical cure rate in group B and group A were statistically significant (P < 0.05), indicating that the two groups of treatment methods all had better results, and group B was better than group A. Conclusion: The senile Brucellosis spondylitis has a characteristic performance. The de-velopment of diagnostic criteria can help to improve the diagnostic rate. Standardized drug treatment has a good cure rate. The implementation of surgery has obvious advantages whether it is to relieve pain, stabilize the spine, restore nerve function, or recover early.

Comparative evaluation of conventional serological tests and in-House recombinant OMP31 (rOMP31)-Based elisa for the detection of caprine brucellosis

Sero-detection of brucella antibodies was done in 300 serum samples randomly collected from aborted goats by the classical serological tests like rose bengal plate test (RBPT), serum tube agglutination test (SAT) and rOMP31-based indirect-Enzyme-Linked Immunosorbent Assay (i-ELISA) and dot-ELISA. The seroprevalence for brucellosis was 22.66, 11.33, 24.33 and 17.33% with RBPT, SAT, i-ELISA and dot-ELISA, respectively. The sensitivity and specificity of different tests varied from 56.25 to 87.50% and from 75.00 to 90.00%, respectively. It was found that rOmp31-based ELISA is the most suitable test with 87.5% sensitivity and 90.0% specificity for detection of brucellosis in goats.

Sero-diagnosis of brucellosis in sheep and humans in Assiut and El-Minya governorates, Egypt

Egypt is an endemic area with brucellosis, so wherever herd problem associated abortion is present, brucellosis should be suspected, and its sero-diagnosis is needed. The present study aimed to estimate the seroprevalence of brucellosis in sheep and their contact humans in Assiut and El-Minya Governorates where a history of abortion in sheep was the chief complaint of the farmers; besides; the appearance of signs of undulant fever among some contact humans. Also, to identify the risk factors for brucellosis seropositivity at human and animal level. Serum samples were collected from 189 sheep and 53 in contact humans in Assiut and El-Minya Governorates, Egypt, during 2017. Antibodies against brucellosis were measured in the serum samples using Rose Bengal Plate test (RBPT) and were further confirmed using Serum Tube Agglutination Test (STAT). The overall seroprevalence ofbrucellosis using RBPT was 15.87% in sheep and that in humans was 9.44% (5 of 53), respectively. STAT confirmed that 40% of the RBPT-positive reactor sheep were infected by Brucella abortus, 16.67% were containing Brucella melitensis and 20% were experiencing a mixed infection of both Br. abortus and Br. melitensis. Additionally, 80% of the RBPT-positive reactor humans were infected with Br. abortus only and 20% were having both species of Brucella. Among different analyzed variables in this study, the age and farming system of the examined sheep were statistically significant. There was no significant effect in relation to gender, abortion history and pregnancy status of the examined animals, however, higher exposure rates were found among males, abortive animals, and recently calved animals. Gender of the tested humans and presence of seropositive sheep in contact differed significantly in acquiring the infection. Nevertheless, other factors, including age, education, and profession were statistically insignificant on getting human brucellosis. These results indicate the high seroprevalence of brucellosis in humans and sheep in the study areas and generally in Egypt, and therefore, control programs should be implemented.

Abortions in an organized dairy farm from North India reveal the possibility of breed susceptibility to Bovine Brucellosis

The present study was undertaken over a three year period (2012–2014) in an organized dairy farm located in North India to ascertain Brucella abortus as the putative cause of abortion. The dairy farm maintained cattle of Frieswal, Crossbred and Sahiwal breeds and followed calf-hood vaccination with Brucella abortus Strain 19 live vaccine in all the heifers. Even with the recommended vaccination schedule and good managemental practices in place, 88 cases of abortions clinically suspected of bovine brucellosis (40 from Frieswal breed, 17 from Crossbred cattle and 31 from Sahiwal breed) were reported from this farm. From these abortion cases, bacteriological isolation was possible in only four dams while 16 dams were found to be serologically positive in Serum Tube Agglutination Test (STAT). Molecular screening by PCR assay (specific for the bcsp31 gene of B. abortus) revealed that 24 dams were positive, out of which 20 were from Frieswal breed and rest four were from Crossbred herd. Prominently, all Sahiwal dams were found to be negative in bacteriological isolation and also in PCR assay. These results thus indicate towards the possibility of breed predisposition to abortions due to B. abortus infection. Statistical analysis by Fischer exact test (p<0.01) too substantiated that breed susceptibility exists among these PCR positive cases. This study is novel as breed variation in abortions due to B. abortus in cattle is being documented for the first time. Seven representative PCR amplicons generated during the study were also sequenced and submitted to NCBI GenBank. Moreover, this study also accentuates the importance of PCR screening especially in vaccinated herd and raises concerns on over-dependence of serological assays when intensive vaccination is practised without any concomitant DIVA strategy. Thus, besides assisting in planning pragmatic control strategies against bovine brucellosis these findings are also imperative from ‘One Health’ context, also.

Comparison of the Rose Bengal Plate and the Complement Fixation Tests with the Tube Agglutination Test for Diagnosis of Human Brucellosis

Brucellosis is a zoonotic disease of economic importance. The clinical manifestations vary in humans; therefore a good diagnostic test is required to confirm the disease. The serum tube agglutination (SA) test, though still the most widely test used, can be problematic for the diagnosis of chronic infections. The other supplementary tests, such as the complement fixation (CF) test and ELISA, require special equipment, reagents and trained personnel. The Rose Bengal plate (RBP) test has shown potential as a good rapid diagnostic test. This is a report of serum samples from suspected cases of brucellosis that were tested using the RBP, SA and CF tests. The RBP test was shown to have a better relative sensitivity and as good specificity as the SA when compared with the CF test, and may be a useful initial diagnostic test for hospitals in remote rural areas if properly conducted with well stored antigen.

Diagnostic role of inflammatory markers in pediatric Brucella arthritis.

BackgroundAs a multisystem infectious disease, there is an inflammation, which causes increase in acute phase reactants in brucellosis. The mean platelet volume (MPV), platelet distribution width (PDW), red cell distribution width (RDW), neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) have been identified as markers of inflammation. The present study aimed to evaluate diagnostic values of these biomarkers in brucella arthritis (BA).MethodsThe study included 64 children with BA and 66 healthy control subjects. Demographic features, joint involvement, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and hematological variables were retrospectively recorded. In addition, results of synovial fluid and serum tube agglutination test for brucella together with treatment regimens were recorded.ResultsThe mean age of the patients (53.1 % male) was 92.3 ± 41.2 months. The most commonly affected joint was ankle (53.1 %). Synovial fluid puncture-brucella agglutination test was positive in 22 (34.3 %) patients. Puncture culture was positive in 9 patients. Most of the patients (57.8 %) were treated with a combination of rifampicin plus sulfamethoxazole/trimethoprim and gentamicin. Significantly higher mean PDW, RDW, MPV, NLR and PLR values were found in children with BA compared to control subjects (p < 0.05). A positive correlation was found between MPV and NLR values (R2 = 0.192, p < 0.001).ConclusionOur findings indicated that NLR and PLR are indirect markers of inflammation that may be observed abnormally increased in children with brucella arthritis. Further longitudinal studies are needed to investigate this topic to establish the more clear associations.

Expansion of brucellosis detection in the country of Georgia by screening household members of cases and neighboring community members.

BackgroundBrucellosis is considered as endemic zoonotic disease in the country of Georgia. However, the burden of the disease on a household level is not known. Therefore, this study sought to determine the benefits of active surveillance coupled to serological screening for the early detection of brucellosis among close contacts of brucellosis cases.MethodsWe used an active surveillance approach to estimate the rate of seropositivity among household family members and neighboring community members of brucellosis index cases. All participants were screened using the serum tube agglutination test (SAT). Blood cultures were performed, obtained isolates were identified by a bacteriological algorithm, and confirmed as Brucella spp. using real-time PCR. Further confirmation of Brucella species was done using the AMOS PCR assay.ResultsA total of 141 participants enrolled. Of these, 27 were brucellosis index cases, 86 were household family members, and 28 were neighboring community members. The serological evidence of brucellosis in the household member group was 7% and the rate at the household level was 21%. No screened community members were Brucella seropositive. Majority of brucellosis cases were caused by B. melitensis; only one index case was linked to B. abortus.ConclusionWe found evidence of brucellosis infection among household family members of brucellosis index cases. B. melitensis was the most common species obtained. Findings of this active surveillance study highlight the importance of screening household family members of brucellosis cases and of the use of culture methods to identify Brucella species in the country of Georgia.

The relationship between mean platelet volume and brucellosis

Dear Editor, I read the article published by Kader et al. [1] with a great interest. They assessed the mean platelet volume (MPV) values in patients with brucellosis. MPV values were significantly lower in the pretreatment group in brucellosis patients. I congratulate the authors for their contribution of the present study, which is successfully written. On the other hand, I want to make minor criticism about this study from methodological aspect. They screened complete blood counts and serum tube agglutination test results retrospectively. However, they didn’t mention about MPV measurement technique in detail which is very important.