National epidemiology of culture-confirmed brucellosis in Israel, 2004-2022.

Brucellosis, a global zoonosis, is endemic in Israel. We used a national database of culture-confirmed cases (2004-2022) to analyse the trends of brucellosis. Of 2,489 unique cases, 99.8% were bacteraemic, 64% involved males, and the mean age was 30.5 years. Brucella melitensis was the dominant species (99.6%). Most cases occurred among the Arab sector (84.9%) followed by the Jewish (8.5%) and Druze (5.5%) sectors. The average annual incidence rates overall and for the Arab, Druze, and Jewish sectors were 1.6/100,000, 6.6/100,000, 5.5/100,000, and 0.18/100,000, respectively. The annual incidence rates among the Arab (incidence rate ratio (IRR) = 36.4) and the Druze (IRR = 29.6) sectors were significantly higher than among the Jewish sector (p < 0.001). The highest incidence rates among the Arab sector occurred in the South District, peaking at 41.0/100,000 in 2012. The frequencies of B. melitensis isolated biotypes (biotype 1 - 69.1%, biotype 2 - 26.0%, and biotype 3 - 4.3%) differed from most Middle Eastern and European countries. A significant switch between the dominant biotypes was noted in the second half of the study period. Efforts for control and prevention should be sustained and guided by a One Health approach mindful of the differential trends and changing epidemiology.

Analysis of the Brucella melitensis epidemic in Xinjiang: genotyping, polymorphism, antibiotic resistance and tracing.

Brucella spp. are facultative intracellular pathogens that cause zoonosis- brucellosis worldwide. There has been a trend of the re-emergence of brucellosis worldwide in recent years. The epidemic situation of brucellosis is serious in Xinjiang. To analyze the epidemic situation of Brucella spp. in Xinjiang among humans and animals, this study identified 144 Brucella isolates from Xinjiang using classical identification and 16 S rRNA sequencing. MLVA, drug resistance testing, and wgSNP detection were also performed. At the same time, analysis was conducted based on the published data of Brucella isolates worldwide. The results showed that the dominant species was B. melitensis biovar 3, which belonged to GT42 (MLVA-8 typing) and the East Mediterranean lineage. The correlation among isolates was high both in humans or animals. The isolates in Xinjiang exhibited higher polymorphism compared to other locations in China, with polymorphism increasing each year since 2010. No amikacin/kanamycin-resistant strains were detected, but six rifampicin-intermediate isolates were identified without rpoB gene variation. The NJ tree of the wgSNP results indicated that there were three main complexes of the B. melitensis epidemic in Xinjiang. Based on the results of this study, the prevention and control of brucellosis in Xinjiang should focus on B. melitensis, particularly strains belonging to B. melitensis bv.3 GT42 (MLVA-8 typing) and East Mediterranean lineage. Additionally, the rifampicin- and trimethoprim-sulfamethoxazole- resistance of isolates in Xinjiang should be closely monitored to avoid compromising the therapeutic efficacy and causing greater losses. These results provide essential data for the prevention and control of brucellosis in Xinjiang and China. Although the isolates from Xinjiang have significant characteristics among Chinese isolates and can reflect the epidemiological situation of brucellosis in China to some extent, this study cannot represent the characteristics of isolates from other regions.

Genomic analysis of Brucella isolates from animals and humans, Türkiye, 2010 to 2020.

BackgroundBrucellosis is a bacterial zoonosis causing severe illness in humans and animals and leading to economic losses in the livestock production in Türkiye and other endemic countries.AimWe aimed at investigating genomic differences of Brucella isolates from animals and humans in Türkiye.MethodsWe used whole genome sequencing (WGS) to assess the genetic diversity of Brucella isolates from 41 provinces in Türkiye and compared with isolates from other countries. We applied allele-based typing and core genome single nucleotide polymorphism (cgSNP) determination.ResultsOf the 106 Turkish Brucella isolates included, 57 were B. abortus and 49 were B. melitensis. One B. melitensis and two B. abortus isolates were identified as vaccine strains. Most (n = 55) B. abortus isolates clustered in three major branches, with no spatial discernible pattern. Of the B. melitensis isolates, 48 were assigned to the Eastern Mediterranean lineage with no discernible patterns between host species, location and sampling date. The Turkish isolates clustered with isolates from neighbouring countries such as Greece and Syria, but some also with isolates from human patients in European countries, like Germany, Norway and Sweden, suggesting that the source may be travel-related.ConclusionSeveral B. melitensis and B. abortus lineages are circulating in Türkiye. To decrease the prevalence and prevent brucellosis in animals and humans, stricter control measures are needed, particularly in areas where humans and animals have close contact. Furthermore, illegal transportation of animals across borders should be more closely controlled and regulated.

[A Case of Thyroid Gland Abscess Caused by Brucella melitensis]. / Brucella melitensis'in Neden Oldugu Bir Tiroid Bezi Apsesi Olgusu.

Brucellosis is a zoonotic disease endemic in many developing countries, including Türkiye. Among the species that are pathogenic for humans; Brucella melitensis is isolated from livestock animals like sheep and goats, Brucella abortus from cattle and Brucella suis from pigs. Laboratory diagnosis of infection caused by Brucella species with gram-negative coccobacillus morphology; can be made through characteristic culture features, serological tests and molecular methods. Brucellosis, which has a wide distribution of clinical signs and symptoms; can cause various complications by affecting many organs and systems. Among all complications, the probability of thyroid abscess is less than 1%. In this case report; an example of thyroid abscess, one of the rare complications of brucellosis that is not frequently encountered in the literature, was presented. During the physical examination of a 45-year-old female patient who admitted with the complaint of pain in the neck area, fever, neck swelling, redness and pain that increased with palpation were detected. Leukocytosis, lymphopenia, high sedimentation and CRP, low TSH and high T4 values were detected in laboratory tests and subacute thyroiditis was considered as the preliminary diagnosis. Surgical abscess drainage was planned as the patient's clinical findings progressed during follow-up and spontaneous pus discharged from the midline of the neck. The abscess aspirate sample taken during surgical intervention and the blood culture samples taken before were evaluated microbiologically. Microorganisms that did not grow on EMB agar but grew on 5% sheep blood and chocolate agar at the 72-96th hour of incubation of culture plates; were detected to have gram-negative coccobacillus morphology and positive for catalase, oxidase and urease. Although the Wright test was negative with a titer of 1/20, the Rose Bengal test was positive, Coombs test was positive with a titer of 1/160 and the Brucellacapt test was positive with a titer of >1/5120. Microorganisms growing on culture plates were identified as B.melitensis at the species level with specific antisera. As a result of antibiotic susceptibility tests evaluated according to the European Committee on Antimicrobial Susceptibility Testing version 14.0 (EUCAST v14.0), the isolate was susceptible to rifampicin, doxycycline, gentamicin and trimethoprim-sulfamethoxazole at standart dosing regimen and susceptible to ciprofloxacin and levofloxacin at increased exposure. The patient, who was started on doxycycline and rifampicin combination treatment, was discharged without any complaints. In the diagnosis of infection due to Brucella species, which is one of the pathogens that early diagnosis and initiation of treatment greatly affects the prognosis; in addition to culture, which is the gold standard method, serological tests are also very important. If diagnosis is delayed, complications may develop due to involvement in almost every part of the body, depending on the affected organs and systems. In areas where brucellosis is endemic, patients with symptoms such as neck swelling, shortness of breath and difficulty in swallowing, thyroid tissue involvement due to brucellosis should definitely be considered etiologically.

Dexamethasone-induced stress exacerbates shedding, tissue antigen distribution, and pathology of caprine Brucellosis.

This study investigated dexamethasone-treatment, shedding routes, tissue antigen distribution, and pathology of caprine Brucellosis. Eighteen non-pregnant goats were randomly grouped into A, B, and C. Group A was administered dexamethasone for 7 days at 2 mg/kg before inoculating 0.5 mL B. melitensis at 107 CFU ocularly while group B was inoculated 0.5 mL B. melitensis only, and C as control negative. Blood samples, ocular, nasal, and vaginal swabs were obtained for evaluation. Three goats were sacrificed from each group at days 21 and 42 post-inoculation (pi) and selected tissues collected for PCR, histopathology, and immunohistochemistry. Brucella melitensis was detected in the ocular swabs of group A significantly higher than group B. Shedding was prolonged in group A compared to B. The overall shedding was 22.2% in group A and 9.4% in group B. The uterus of both groups A and B revealed mild inflammation and microgranuloma, extensive necrotic lesions in lymph nodes. Liver showed multifocal necrosis predominantly in group A. Lesion scoring showed significantly higher scores in A compared to B. Strong immunostaining was observed in the liver, lungs, and spleen, predominantly at day 21 pi. This study demonstrated dexamethasone prolonged shedding, tissue antigen distribution, and pathology in dexamethasone-treated goats.

Meningoencephalitis, coronary artery and keratitis as an onset of brucellosis: a case report.

BACKGROUND: Brucellosis is a zoonotic disease caused by brucella. It has been an increasing trend in recent years (Wang H, Xu WM, Zhu KJ, Zhu SJ, Zhang HF, Wang J, Yang Y, Shao FY, Jiang NM, Tao ZY, Jin HY, Tang Y, Huo LL, Dong F, Li ZJ, Ding H, Liu ZG, Emerg Microbes Infect 9:889-99, 2020). Brucellosis is capable to invade multiple systems throughout the body, lacking in typical clinical manifestations, and easily misdiagnosed and mistreated. CASE PRESENTATION: We report a case of a male, 5-year-and-11-month old child without relevant medical history, who was admitted to hospital for 20 days of fever. When admitted to the hospital, we found that he was enervated, irritable and sleepy, accompanied with red eyes phenomenon. After anti-infection treatment with meropenem, no improvement observed. Lumbar puncture revealed normal CSF protein, normal cells, and negative culture. Later, doppler echocardiography suggested coronary aneurysms, and incomplete Kawasaki Disease with coronary aneurysms was proposed. The next day, brucellosis agglutination test was positive. Metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid suggested B.melitensis, which was confirmed again by blood culture. The child was finally diagnosed as brucellosis with meningocephalitis, coronary aneurysm and keratitis. According to our preliminary research and review, such case has never been reported in detail before. After diagnosis confirmation, the child was treated with rifampicin, compound sulfamethoxazole, and ceftriaxone for cocktail anti-infection therapy. Aspirin and dipyridamole were also applied for anticoagulant therapy. After medical treatment, body temperature of the child has reached normal level, eye symptoms alleviated, and mental condition gradually turned normal. Re-examination of the doppler echocardiographic indicated that the coronary aneurysm was aggravated, so warfarin was added for amplification of anticoagulation treatment. At present, 3 months of follow-up, the coronary artery dilatation gradually assuaged, and the condition is continued to alleviate. CONCLUSION: Brucellosis can invade nervous system, coronary artery, and cornea. Brucellosis lacks specific signs for clinical diagnosis. The traditional agglutination test and the new mNGS are convenient and effective, which can provide the reference for clinical diagnosis.

Brucella melitensis B115-based ELISA to unravel false positive serologic reactions in bovine brucellosis: a field study.

BACKGROUND: Brucellosis is a zoonosis whose incidence is not declining worldwide despite the global effort to control the disease. Accurate and precise diagnosis is a crucial step in any prophylaxis program but single tests to unequivocally detect animals infected with Brucella spp. are currently unavailable. In Italy, serological diagnosis of bovine brucellosis is performed with two official tests: a rapid agglutination test (i.e., Rose Bengal Plate test, RBPT) and a complement fixation test (CFT) that detect antibodies directed mainly to the smooth lipopolysaccharide (S-LPS). Neither of the two tests is able to avoid the detection of false positive serological reactions (FPSRs) caused by bacteria sharing S-LPS components with Brucella spp. and responsible for the single reactors (SR) phenomenon. A B. melitensis R strain-based ELISA showed a good diagnostic performance in unravelling FP animals; however, since a limited number of animals were analyzed in that study, a large field study was conducted here to discriminate between Brucella-infected from FP animals, with the final aim of reducing the unnecessary slaughter of the latter. An ELISA based on a R strain of Brucella, i.e., Brucella melitensis B115, was employed to measure specific IgG responses in a collection of bovine sera (n = 648). Sera were obtained from 180 farms (either officially brucellosis-free or not brucellosis-free) recruited during an extended period of time (2014-2018) and were preliminarily assayed with the official tests by the Italian Reference Centers and then subjected to the ELISA. RESULTS: Negative sera, when subjected to the ELISA, gave O.D. values below the cutoff; SR sera, i.e. RBPT positive and CFT negative, as well as double positive (DP) sera, i.e. RBPT and CFT positive, gave O.D. values that were below the cutoff. All positive sera, i.e. from Brucella-infected animals, were RBPT positive and CFT positive (ICFTU ranging from 20 to 1280) and gave ELISA O.D. values above the cutoff. CONCLUSIONS: The B. melitensis B115-based ELISA systematically unravelled all false positive (FP) sera while confirming the diagnosis in Brucella-infected animals. Thus, the test employed in the present study may complement the official assays to avoid the costly slaughter of FP animals.

Isolation and identification of Brucella melitensis using bacteriological and molecular tools from aborted goats in the Afar region of north-eastern Ethiopia.

BACKGROUND: Infection with Brucella melitensis (B. melitensis) is one of the most important causes of abortion in goats and sheep, and also causes severe systemic disease in exposed humans. In Ethiopia, based on seroepidemiological studies, brucellosis is known to be endemic. However, there is little information on the isolation and molecular detection of Brucella species in small ruminants. Therefore, the present study was conducted in the Amibara district of Afar Region of Ethiopia to isolate and molecularly detect Brucella infection in small ruminants. RESULTS: Out of the total 64 samples cultured, eight samples (five vaginal swabs and three milk) were positive for Brucella species based on colony morphology, growth characteristics, modified acid fast staining and biochemical tests results. Further identification using Brucella- ladder PCR method showed that four of the isolates (three from vaginal swabs and one from milk) from goats amplified fragments of 1071 bp, 794 bp, 587 bp, 450 bp and 152 bp in band size. The molecular result combined with the microbiological and biochemical characteristics of the isolates indicated that the isolates were strains of B. melitensis. CONCLUSION: The finding of this study could suggest economic and zoonotic significance of B. melitensis and warrants for the need for control strategies in livestock and creation of awareness in the pastoral communities on the safe consumption of foods of animal origin and avoidance of physical contact with aborted materials.

Fluorescence signal amplification assay for the detection of B. melitensis 16M, based on peptide-mediated magnetic separation technology and a AuNP-mediated bio-barcode assembled by quantum dot technology.

Members of the Brucella spp. are facultative intracellular bacteria that can cause global brucellosis, a zoonotic disease. Herein, a novel fluorescence signal amplification (FSA) method for the rapid detection of B. melitensis 16M was developed based on peptide-mediated magnetic separation (PMS) technology and Au nanoparticle (AuNP)-mediated bio-barcode assay technology assembled by quantum dots (QDs). The PMS technology was used to specifically capture and isolate B. melitensis 16M from food. The immunomagnetic bead-B. melitensis 16M bioconjugates (IMBs-B. melitensis 16M) were then identified by IgY on the surface of AuNPs and the oligonucleotide chains on the surface of the gold nanoparticles were hybridized with bio-barcodes assembled by quantum dots (QD-probe2). The IMB/B. melitensis 16M/IgY-AuNP-probe1/QD-probe2 bioconjugates were concentrated by magnetic separation. Therefore, as the concentration of B. melitensis 16M in the sample increased, the unbound QD-probe2 in the supernatant reduced, and the B. melitensis 16M in the sample could be indirectly measured by detecting the fluorescence in the supernatant. This FSA method can detect B. melitensis 16M concentration in the range of 10 to 106 cfu ml-1 without pre-enrichment, and the limit of detection (LOD) is as low as 10 cfu ml-1 with high specificity. Furthermore, the proposed method for the detection of B. melitensis 16M has a LOD of 1.07 × 102 cfu ml-1 and a linear range from 102 to 107 cfu ml-1 in milk, and a LOD of 1.72 × 102 cfu ml-1, and a linear range from 102 to 106 cfu ml-1 in lamb leach. In addition, this method takes less than 3 h to perform. Thus, the assay that was developed in this study shows promise for rapid, sensitive, and specific detection of B. melitensis 16M.

A case report on mother-to-child transmission of Brucella in human, China.

BACKGROUND: Human brucellosis is endemic in China and commonly occurs through contact with infected animals from working with livestock or consumption of unpasteurized dairy products. Although rare, human-to-human, and possible sexual transmission, of Brucella has been reported. In this report, we describe a case of likely mother-to-child transmission of Brucella in Hunan Province, China. CASE PRESENTATION: Between June and October 2016, a 28-year old man sought care for testicular swelling and pain at several health facilities. His 26-year old wife developed intermittent fever along with right thigh and hip pain between November 2016 and February 2017 respectively. On April 5, 2017, the female patient delivered a male neonate at 34 weeks of gestation through natural labor. The child's venal blood sample was cultured on April 5, 2017. Brucella was isolated and identified on April 12, 2017. On the same date, serum antibodies of the father and mother were above 1:100 (based on the serum agglutination test [SAT]). The strains isolated from the mother and neonate were identified as Brucella melitensis biotype 1. CONCLUSIONS: This report highlights a family cluster of brucellosis. Culture results strongly support mother-to-child transmission, and a high probability of sexual transmission from husband to wife.

Associated factors in distinguishing patients with brucellosis from suspected cases.

BACKGROUND: To investigate the risk factors for brucellosis in suspected cases of the disease. METHODS: A self-designed questionnaire was developed to collect data from 3557 people whose initial visit site was the Songyuan Center for Disease Control and Prevention (CDC) from January 1st, 2009 to December 31st, 2012. After collecting blood samples, a plate agglutination test (PAT) and serum agglutination test (SAT) were used to distinguish the patients with brucellosis from the suspected cases. RESULTS: Sex, occupation (farmers and herdsmen), contact with abortion products, and contact with feces were the main risk factors for brucellosis in the suspected cases (all P < 0.05). No difference existed between the confirmed cases and suspected cases in the demographic characteristics, contact with animals (except swine), contact with substances, or clinical symptoms (except fever). However, the confirmed cases showed significant differences from people without brucellosis in demographic characteristics, contact with animals (except cattle and swine), contact with substances, and clinical symptoms. Suspected cases exhibited significant differences from people without brucellosis in the demographic characteristics (except education), contact with animals (except swine), contact with substances (except dust), and clinical symptoms (except chills and acratia). Brucella was cultured from the blood samples of three of 30 suspected cases with fever. Using AMOS-PCR and agarose electrophoresis, the detailed species of Brucella strain was identified as Brucella melitensis. CONCLUSIONS: Abortion products and feces are the main risk factors for brucellosis in suspected cases of the disease. Pyrexia in suspected cases with a history of contact with abortion products or feces should raise suspicion for the disease.

Hip osteoarticular complication due to delay in diagnosis and treatment of brucellar hip arthritis: two cases report.

BACKGROUND: Brucellosis is a systemic infectious disease frequently associated with osteoarticular involvement. While sacroiliitis is a common manifestation of brucellosis, septic osteoarthritis is less frequent. Here, we report two cases of septic osteoarthritis caused by Brucella melitensis. CASE PRESENTATION: Both patients had a history of contact with goats before admission. Upon clinical examination, they showed marked pain and limited movement in the hip. Imaging findings revealed obvious osteonecrosis of the right femoral head. Inflammatory markers, including erythrocyte sedimentation rate and C-reactive protein level, were elevated. The tube agglutination test results of both patients were positive (1:160 and 1:200). Real-time polymerase chain reaction analysis of synovial fluid revealed the presence of B. melitensis. We deduced that septic osteoarthritis could explain these clinical and radiological findings. Both patients were followed-up for 12 months. They returned to their normal routine after completing a standard antibiotic regimen, including doxycycline (100 mg, daily) and rifampicin (600 mg), for 6 weeks. CONCLUSIONS: Brucellar hip arthritis is a serious clinical manifestation of brucellosis, presenting mainly as marked joint pain and limited mobility. It is characterized by joint effusion, synovitis, and soft-tissue swelling on magnetic resonance images. Physicians should consider brucellosis as one of the differential diagnoses of arthritis.

Diagnostic utility of in-house loop-mediated isothermal amplification and real-time PCR targeting virB gene for direct detection of Brucella melitensis from clinical specimens.

AIMS: In this present study, the utility of a newly developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. METHODS AND RESULTS: Fifty-four culture-confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real-time PCR. Overall sensitivities of LAMP and real-time PCR assays were 67·5 and 68·3% respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9%) and real-time PCR (100%) assays. CONCLUSIONS: Performance of LAMP and real-time PCR was not satisfactory for whole-blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed LAMP and real-time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.

Rare Meningitis and Epileptic Seizure Infected with Brucella melitensis: a Case Report.

BACKGROUND: Although brucellosis is the most common zoonotic disease worldwide, human meningitis infected with Brucella melitensis is rare and difficult to diagnosis. Herein we describe the clinical aspects of a rare case of Brucella melitensis meningitis accompanied by epileptic seizure. METHODS: Bacterial culture of CSF was utilized to find the pathogen. Serum and CSF agglutination tests were used to detect the capacity of Brucella antigen. Bacterial clone was identified by the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF). Magnetic resonance imaging (MRI) was used to monitor the recovery changes for meningitis. RESULTS: The bacterial culture was positive for Brucella app. Antigen-antibody agglutination test was also positive with a titer more than 1/200. A reliable identification score of 2.8 for Brucella melitensis was obtained by MALDI-TOF. MRI showed obviously improved meningitis after therapy. CONCLUSIONS: The results of this study suggest the treatments for Brucella meningitis must be planned depending on the patient's clinical conditions and the laboratory identification of pathogen infection as early as possible.

Successful Endovascular Aneurysm Repair for Brucella Mycotic Aneurysm Acquired from Ingestion of Sheep Placenta.

An aortic aneurysm infected by Brucella is rarely seen. In this case report, we describe a Brucella mycotic abdominal aortic aneurysm acquired from ingestion of sheep placenta. Clinical symptoms included fever, fatigue, and abdominal pain. Diagnosis was confirmed by blood culture and computed tomography (CT) angiography. The patient had increased abdominal pain during hospitalization, and repeated CT showed a rapidly enlarging mycotic aneurysm. Emergent endovascular repair was successfully performed using a bifurcated stent graft, and combined intravenous and oral antibiotics were administrated. The patient was asymptomatic after operation, and follow-up CT showed thrombosis in the aneurysmal sac and significant decrease of aneurysmal size.

Brucellosis as a rare cause of olecranon bursitis: case-based review.

A 51-year-old man shepherd presented with mild pain and swelling of the right posterior aspect of his right elbow. In ultrasonography, the affected bursal space had swelling and effusion. Moreover, the aspiration of the affected bursa revealed an inflammatory profile. Brucella melitensis was detected in aspirated fluid and blood cultures. The serum agglutination test (SAT) and 2-mercaptoethanol test for brucellosis were also positive. Therefore, the diagnosis of brucellar olecranon was confirmed. Treatment was initiated using gentamicin for the first 7 days and doxycycline plus rifampicin for 2 months. After treatment, all clinical signs and symptoms were resolved. No relapse was seen after 1 year of the completion of treatment. Clinicians should pay attention to the symptoms of olecranon brucellar bursitis that is similar to that of pyogenic bursitis.

Phenotypic and genotypic characteristics of Brucella isolates from the Republic of Kazakhstan.

The purposes of this study were to determine phenotypic and genotypic characteristics of Brucella isolates from the Republic of Kazakhstan and to determine their biotype. The focus was laid on culture-morphological, biochemical, and biological properties of 59 Brucella isolates from primary cultures. Material was isolated from blood and tissue of serum-positive killed, dead diseased, or aborted domestic cattle from different regions of Kazakhstan where brucellosis is a common problem. Multiple-locus variable number tandem repeat analysis (MLVA) of all strains, isolated in different regions, has shown that Brucella isolates from the epizootic form two clusters. Based on the comparison with strains available in the MLVA database, B. abortus 0015/B is alike the B. abortus strains isolated from Italy and Portugal. B. melitensis 0016/B isolated from the Almaty region fits the third cluster and is alike the B. melitensis strains isolated from humans in Turkey, China, and Portugal. More than 90% of the overall B. abortus samples were isolated from the northern regions of the East and West Kazakhstan, while B. melitensis strains were registered in the southeast Kazakhstan. The most frequently recorded B. abortus biovar is biovar 3. The most frequently recorded B. melitensis biovars are biovars 1 and 3. SIGNIFICANCE AND IMPACT OF STUDY: These results contribute to a better understanding of the geographic pattern of Brucella infection in Kazakh cattle also important for developing the specific control measures. The results of current research can be used for creating a gene bank of Brucella strains circulating in Kazakhstan for producing diagnostic and therapeutic agents. The research material will be used to solve the problems of genetic characterization of Brucella species and to establish the phylogenetic relationships of strains.

In vitro antimicrobial susceptibility testing of human Brucella melitensis isolates from Ulanqab of Inner Mongolia, China.

BACKGROUND: Brucellosis is an endemic disease in the Inner Mongolia Autonomous Region of China and Ulanqab exhibits the highest prevalence of brucellosis in this region. Due to the complex nature of Brucellosis, a cure for this disease has proven to be elusive. Furthermore, the reduced susceptibility of Brucella spp. to antimicrobial agents has been reported as a potential cause of therapeutic failure. However, detailed in vitro antimicrobial susceptibility patterns pertaining to Brucella isolates from this region have not yet been published. The aim of this study was to evaluate the antibiotic susceptibility profile of Brucella melitensis clinical isolates from Ulanqab, Inner Mongolia, China. METHODS: A total of 85 B. melitesis isolates were obtained from humans in Ulanqab of Inner Mongolia, China; the antimicrobial susceptibility of 85 clinical isolates to nine antibiotics was assessed using the E-test method according to the CLSI (Clinical and Laboratory Standards Institute) guidelines. RESULTS: All of the tested isolates were susceptible to minocycline, sparfloxacin, doxycycline, tetracycline, ciprofloxacin, gentamicin and levofloxacin. Resistance to rifampin and cotrimoxazole was observed in 1.0% (1/85) and 7.0% (6/85) of the isolates, respectively. However, rpoB gene mutations were not observed in single isolates exhibiting resistance to rifampin. CONCLUSIONS: We observed that B. melitensis isolates are susceptible to the majority of the tested antibiotics. Furthermore, minocycline and sparfloxacin exhibited extremely high bactericidal effects in relation to the B. melitensis isolates. The sensitivity of commonly used drugs for the treatment of brucellosis should be regularly monitored. To the best of our knowledge, this is the first report of rifampin and cotrimoxazole resistant isolates of B. melitensis in China. In summary, based on the findings from this study, we suggest that antibiotic administration and use should be rationalized to prevent future drug resistance.

Documenting the absence of brucellosis in cattle, goats and dogs in a "One Health" interface in the Mnisi community, Limpopo, South Africa.

This study shows the absence of the world's most common bacterial zoonoses caused by Brucella abortus and Brucella melitensis in cattle, goats and dogs in an agro-pastoral community in South Africa, where heifer vaccination against brucellosis with the live Strain 19 vaccine is compulsory. The study site is bordering wildlife reserves with multiple wildlife species infected with brucellosis. The results showed a low seroprevalence (1.4%) in cattle. Seroprevalence in cattle decreased with age after 4 years in females, males were less positive than females and a tissue culture from a brucellin skin test-positive male was negative. The results indicate that Brucella seropositivity in cattle is due to S19 vaccination and not natural infections. This conclusion is reinforced by the absence of Brucella seropositivity in goats (1/593 positive result) and dogs (0/315), which can be seen as potential spillover hosts. Therefore, the close proximity of brucellosis-infected wildlife is not a threat to domestic animals in this controlled setting with vaccination, fencing and movement control.

Brucellosis in migratory sheep flock from Maharashtra, India.

Brucellosis is a zoonotic disease worldwide distributed and having the economic as well as public health importance. The prevalence of brucellosis among sheep flock having history of abortions was studied. A total of 229 samples comprising of 157 blood and 72 clinical samples (vaginal swabs) were collected from 157 animals. Clinical samples were processed for the isolation of Brucella melitensis. Serum samples (n = 157) were tested by Rose Bengal plate test (RBPT) and i-ELISA. A total of 68 (43.31%) and 104 (66.24%) samples were positive by RBPT and ELISA, respectively. Brucella isolates (n = 2) were recovered from clinical samples. Both isolates demonstrated amplification for bcsp 31 and IS711 genes. On AMOS PCR, both the isolates amplified at 731 bp, i.e., belongs to B. melitensis species. The incidence of B. melitensis in a migratory flock warns the thorough testing and culling of Brucella-infected sheep from the flock on a continuous basis; otherwise, such incidence will be routine and poor farmers will be at a loss.