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protective live oral brucellosis vaccines stimulate th1 and th17 cell responses.zoonotic transmission of brucellosis often results from exposure to brucella-infected livestock, feral animals, or wildlife, or frequently via consumption of unpasteurized milk products or raw meat. since natural infection of humans often occurs by the oral route, mucosal vaccination may offer a means to confer protection for both mucosal and systemic tissues. significant efforts have focused on developing a live brucellosis vaccine, and deletion of znua gene involved in zinc transport has been ...201121768283
cutaneous manifestations in brucellosis.brucellosis is a common worldwide zoonotic disease. cutaneous manifestations are not specific and affect 1-14% of patients with brucellosis. here, we describe 49-year-old female with fever and a diffuse maculopapular rash due to brucella melitensis infection. histopathology of skin biopsy revealed leukocytoclastic vasculitis; positive blood cultures for b. melitensis established the diagnosis of brucellosis. we provide a review of the relevant literature.201121772606
[genetic characterization of the brucella melitensis isolates from mongolia, russia, and azerbaijan].the goal of this work was to provide comparative genetic characterization of the human and animal brucella melitensis isolates from mongolia, russia and azerbaijan using current molecular-genetic typing methods.201121786630
mlva genotyping of human brucella isolates from turkey.a multiple loci variable-number tandem-repeat analysis (mlva) was applied to investigate the epidemiological relationship and genetic diversity among 162 human brucella isolates collected from all geographic regions of turkey in an 8-year period (2001-2008). the isolates were genotyped by using mlva-16(upsud) including eight minisatellite (panel 1) and eight microsatellite (panel 2 subdivided in 2a and 2b) markers. panels 1 and 2a distinguish 14 genotypes; two of these represented 85% of the str ...201121795514
transcriptional profile of the intracellular pathogen brucella melitensis following hela cells infection.brucella spp. infect hosts primarily by adhering and penetrating mucosal surfaces; however the initial molecular phenomena of this host:pathogen interaction remain poorly understood. using cdna microarray analysis, we characterized the transcriptional profile of the intracellular pathogen brucella melitensis at 4h (adaptational period) and 12h (replicative phase) following hela cells infection. the intracellular pathogen transcriptome was determined using initially enriched and then amplified b. ...201121798337
brucella melitensis infection following military duty in iraq.brucellosis is a common zoonotic disease worldwide; however, few cases are reported in the us. brucella melitensis infections are primarily acquired via consumption of high-risk foods or travel to endemic areas. we describe a case of b. melitensis infection in a tennessee soldier following deployment in iraq. initial symptoms included knee and back pain. culture of an aspirate of the left sacroiliac joint yielded b. melitensis. genetic analysis indicated that this isolate came from the middle ea ...201121824352
epidemiological survey of rifampicin resistance in clinic isolates of brucella melitensis obtained from all regions of turkey.the aim of the present study was to assess the antimicrobial susceptibility of brucella melitensis isolates to rifampicin (rif) depending on time and regional differences. a total of 94 human brucella isolates collected in an 8-year period from the beginning of 2002 to the end of 2009 throughout turkey were investigated. the isolates were identified at species and biovar levels by conventional methods, and minimum inhibitory concentrations (mic) of rif was determined by using the e test method. ...201121826587
Brucella melitensis cyclic di-GMP phosphodiesterase BpdA controls expression of flagellar genes.Brucella melitensis encounters a variety of conditions and stimuli during its life cycle--including environmental growth, intracellular infection, and extracellular dissemination--which necessitates flexibility of bacterial signaling to promote virulence. Cyclic-di-GMP is a bacterial secondary signaling molecule that plays an important role in adaptation to changing environments and altering virulence in a number of bacteria. To investigate the role of cyclic-di-GMP in B. melitensis, all 11 pred ...201121856843
systems biology approach predicts antibody signature associated with brucella melitensis infection in humans.a complete understanding of the factors that determine selection of antigens recognized by the humoral immune response following infectious agent challenge is lacking. here we illustrate a systems biology approach to identify the antibody signature associated with brucella melitensis (bm) infection in humans and predict proteomic features of serodiagnostic antigens. by taking advantage of a full proteome microarray expressing previously cloned 1406 and newly cloned 1640 bm genes, we were able to ...201121863892
structure of fructose bisphosphate aldolase from bartonella henselae bound to fructose 1,6-bisphosphate.fructose bisphosphate aldolase (fbpa) enzymes have been found in a broad range of eukaryotic and prokaryotic organisms. fbpa catalyses the cleavage of fructose 1,6-bisphosphate into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. the ssgcid has reported several fbpa structures from pathogenic sources, including the bacterium brucella melitensis and the protozoan babesia bovis. bioinformatic analysis of the bartonella henselae genome revealed an fbpa homolog. the b. henselae fbpa enzym ...201121904049
BrabA.11339.a: anomalous diffraction and ligand binding guide towards the elucidation of the function of a 'putative ß-lactamase-like protein' from Brucella melitensis.The crystal structure of a ß-lactamase-like protein from Brucella melitensis was initially solved by SAD phasing from an in-house data set collected on a crystal soaked with iodide. A high-resolution data set was collected at a synchroton at the Se edge wavelength, which also provided an independent source of phasing using a small anomalous signal from metal ions in the active site. Comparisons of anomalous peak heights at various wavelengths allowed the identification of the active-site metal i ...201121904058
Solution structure of an arsenate reductase-related protein, YffB, from Brucella melitensis, the etiological agent responsible for brucellosis.Brucella melitensis is the etiological agent responsible for brucellosis. Present in the B. melitensis genome is a 116-residue protein related to arsenate reductases (Bm-YffB; BR0369). Arsenate reductases (ArsC) convert arsenate ion (H(2)AsO(4)(-)), a compound that is toxic to bacteria, to arsenite ion (AsO(2)(-)), a product that may be efficiently exported out of the cell. Consequently, Bm-YffB is a potential drug target because if arsenate reduction is the protein's major biological function t ...201121904062
comparative analysis of glutaredoxin domains from bacterial opportunistic pathogens.glutaredoxin proteins (glxrs) are essential components of the glutathione system that reductively detoxify substances such as arsenic and peroxides and are important in the synthesis of dna via ribonucleotide reductases. nmr solution structures of glutaredoxin domains from two gram-negative opportunistic pathogens, brucella melitensis and bartonella henselae, are presented. these domains lack the n-terminal helix that is frequently present in eukaryotic glxrs. the conserved active-site cysteines ...201121904064
limitations of the standard agglutination test for detecting patients with brucella melitensis bacteremia.abstract the sensitivity of the standard agglutination test (sat) for detecting brucellosis was determined in 264 israeli patients from whom a positive blood culture for brucella melitensis and serology were obtained within±1 week. a sat titer ≥1:160 had a diagnostic sensitivity of 91.7%, whereas raising the cutoff to ≥1:320, as recommended to decrease false-positive rates in endemic areas, reduced the sensitivity to 82.6%. physicians working in regions endemic for brucellosis should be aware ...201121919731
identification of immunoreactive proteins of brucella melitensis by immunoproteomics.infection with brucella causes brucellosis, a chronic disease in humans, which induces abortion and sterility in livestock. among the different brucella species, brucella melitensis is considered the most virulent and is the predominant species associated with outbreaks in china. to date, no safe human vaccine is available against brucella infection. the currently used live vaccines against brucella in livestock induce antibodies that interfere with the diagnosis of field infection in vaccinated ...201121922434
[comparison of conventional methods and real-time multiplex polymerase chain reaction for identification and typing of brucella isolates of human origin].brucellosis which is a worldwide zoonotic disease, still constitutes a major public health problem in rural areas of turkey. the aim of the present study was to evaluate the species and biovar distribution of 187 presumptive brucella strains isolated from patients inhabiting at the provinces in eastern, south eastern and mediterranean regions over a 7-years period (from 2001 to 2007) and to compare multiplex real-time-polymerase chain reaction (m-rt-pcr) and conventional biotyping for the differ ...201121935772
Interactions of the human pathogenic Brucella species with their hosts.Brucellosis is a zoonotic infection caused primarily by the bacterial pathogens Brucella melitensis and B. abortus. It is acquired by consumption of unpasteurized dairy products or by contact with infected animals. Globally, it is one of the most widespread zoonoses, with 500,000 new cases reported each year. In endemic areas, Brucella infections represent a serious public health problem that results in significant morbidity and economic losses. An important feature of the disease is persistent ...201121939378
Immunoproteomic analysis of Brucella melitensis and identification of a new immunogenic candidate protein for the development of brucellosis subunit vaccine.In order to screen immunogenic candidate antigens for the development of a brucellosis subunit vaccine, an immunoproteomic assay was used to identify immunogenic proteins from Brucella melitensis 16 M soluble proteins. In this study, a total of 56 immunodominant proteins were identified from the two-dimensional electrophoresis immunoblot profiles by liquid chromatography tandem mass spectrometry (LC-MS/MS). Two proteins of interest, riboflavin synthase alpha chain (RS-a) and Loraine synthase (LS ...201121943783
Survey to determine the seroprevalence of Toxoplasma gondii infection in British sheep flocks.A serological survey of Toxoplasma gondii infection in adult breeding sheep in Great Britain was conducted using surplus sera taken during a seroprevalence study of Brucella melitensis in 2009. Of the 3539 sera collected from 227 flocks, 2619 (74 per cent) were found to be positive for T gondii specific antibody when tested using latex agglutination. Multilevel logistic modelling suggested that the likelihood of infection increased with age and this effect appeared to be amplified in animals vac ...201121957115
brucella melitensis infection within warthin tumor of the parotid gland.brucellosis is a zoonotic systemic infectious disease, and multiorgan involvement is commonly seen, but involvement of the neck is a rare presentation of brucellosis. granulomatous infections of the parotid gland are extremely rare. warthin tumor is a well-known benign neoplasm of the salivary glands. in this report, we describe a warthin tumor associated with brucella melitensis in the same parotid gland.201121959459
roles of genomic island 3 (gi-3) bab1_0278 and bab1_0263 open reading frames (orfs) in the virulence of brucella abortus in balb/c mice.the genomic island 3 (gi-3) shared by brucella melitensis and brucella abortus contains 29 genes encoding mostly unknown proteins. within this island, the open reading frames (orfs) bab1_0278 and bab1_0263 are present, bab1_0278 encodes a hypothetical protein of 64 amino acids sharing a domain with the gcra superfamily, whereas the amino acid sequence of bab1_0263 showed 42% identity with an iron regulated lsr2 protein. we obtained one deletion mutant for each one of these orfs present within th ...201122005180
Molecular typing of Brucella melitensis endemic strains and differentiation from the vaccine strain Rev-1.In the present study forty-four Greek endemic strains of Br. melitensis and three reference strains were genotyped by Multi locus Variable Number Tandem Repeat (ML-VNTR) analysis based on an eight-base pair tandem repeat sequence that was revealed in eight loci of Br. melitensis genome. The forty-four strains were discriminated from the vaccine strain Rev-1 by Restriction Fragment Length Polymorphism (RFLP) and Denaturant Gradient Gel Electrophoresis (DGGE). The ML-VNTR analysis revealed that en ...201122021041
long-term survival of salvage cardiac transplantation for infective endocarditis.cardiac transplantation has been rarely performed in patients with infective endocarditis. a 31-year-old man developed aortic endocarditis due to brucella melitensis. he presented with fever and developed acute myocardial infarct, severe aortic regurgitation, and heart failure. aortic valve replacement did not improve cardiac function; hence, an emergent cardiac transplantation was carried out. eighteen years later, he is doing well and living an active and productive life. only 6 patients have ...201122051318
pathophysiology of the rhesus macaque model for inhalational brucellosis.the objective of this study was to characterize the rhesus macaque (rm) as a model for inhalational brucellosis in support of the u.s. food and drug administration's (fda) animal rule. the pathophysiology of chronic brucella melitensis aerosol infection was monitored in two phases that each occurred over an 8-week time period; dose escalation (8 rms; targeted doses of 5.0e+03, 5.0e+04, or 5.0e+05 cfu/animal or the unchallenged control) and natural history (12 rms; targeted dose of 2.50e+05 cfu/a ...201222064715
mlva genotyping of chinese human brucella melitensis biovar 1, 2 and 3 isolates.since 1950, brucella melitensis has been the predominant strain associated with human brucellosis in china. in this study we investigated the genotypic characteristics of b. melitensis isolates from china using a multiple-locus variable-number tandem-repeat analysis (mlva) and evaluated the utility of mlva with regards to epidemiological trace-back investigation.201122108057
Brucella melitensis Survival During Manufacture of Ripened Goat Cheese at Two Temperatures.Abstract The aim of the current work was to assess the influence of two temperatures, 4°C and 24°C, on pH and water activity and their association with Brucella melitensis survival during the traditional manufacture of ripened goat cheese. Raw milk from a brucellosis-free goat herd was used for the manufacture of ripened cheese. The cheese was inoculated with 5×10(9) of the B. melitensis 16M strain during the tempering stage. The cheeses were matured for 5, 20, and 50 days at both temperatures ...201122129442
acute brucellosis with myelodysplastic syndrome presenting as pancytopenia and fever of unknown origin.objective: to report a rare case of brucellosis with myelodysplastic syndrome (mds). clinical presentation and intervention: a 70-year-old woman presented with pancytopenia and fever of unknown origin (fuo). the initial diagnosis was brucellosis; the woman was treated with doxycycline and rifampin against brucella melitensis but was later diagnosed as suffering from mds. she was immediately transferred to the department of hematology for further evaluation. conclusion: this study highlights the ...201122133749
Acellular vaccines for ovine brucellosis: a safer alternative against a worldwide disease.Ovine brucellosis is a very contagious zoonotic disease distributed worldwide and constitutes a very important zoosanitary and economic problem. The control of the disease includes animal vaccination and slaughter of infected flocks. However, the commercially available vaccine in most countries is based on the attenuated strain Brucella melitensis Rev 1, which presents important safety drawbacks. This review is focused on the most recent and promising acellular vaccine proposals.201222149711
The Brucella TIR-like protein TcpB interacts with the death domain of MyD88.The pathogen Brucella melitensis secretes a Toll/interleukin-1 receptor (TIR) domain containing protein that abrogates host innate immune responses. In this study, we have characterized the biochemical interactions of Brucella TIR-like protein TcpB with host innate immune adaptor proteins. Using protein-fragment complementation assays based on Gaussia luciferase and green fluorescent protein, we find that TcpB interacts directly with MyD88 and that this interaction is significantly stronger than ...201122155231
extended safety and efficacy studies of the attenuated brucella vaccine candidates 16m{delta}vjbr and s19{delta}vjbr in the immunocompromised irf-1-/- mouse model.the global distribution of brucellosis and high incidence in certain areas of the world warrants the development of a safer and efficacious vaccine. for the past ten years, we have focused our attention on the development of a safer, but still highly protective, live-attenuated vaccine for human and animal use. we have demonstrated the safety and protective efficacy of the vaccine candidates 16mδvjbr and s19δvjbr against homologous and heterologous challenge in multiple immunocompetent animal mo ...201122169089
Acute renal failure due to Brucella melitensis.We present the case of a 42-year-old male patient who applied to the emergency department of our hospital with clinical nephritis, orchitis, acute renal failure without endocarditis, and a low-grade fever. Brucella agglutinin titers were 1:160, Rose Bengal test was positive and Brucella melitensis was isolated from urine and blood cultures.. A combination of oral rifampin (600 mg/day) and doxycycline (200 mg/day) was administered along with supportive treatment leading to resolution of his clini ...201122169790
simultaneous detection and differentiates of brucella abortus and brucella melitensis by combinatorial pcr.to evaluate simultaneous detection and differentiates of brucella abortus (b. abortus) and brucella melitensis (b. melitensis) through the combinatorial pcr method.201222182638
deep-sequencing analysis of the mouse transcriptome response to infection with brucella melitensis strains of differing virulence.brucella melitensis is an important zoonotic pathogen that causes brucellosis, a disease that affects sheep, cattle and occasionally humans. b. melitensis strain m5-90, a live attenuated vaccine cultured from b. melitensis strain m28, has been used as an effective tool in the control of brucellosis in goats and sheep in china. however, the molecular changes leading to attenuated virulence and pathogenicity in b. melitensis remain poorly understood. in this study we employed the illumina genome a ...201122216095
serial kinetics of the antibody response against the complete brucella melitensis orfeome in focal vertebral brucellosis.human brucellosis is a common zoonosis worldwide. here we present a case of focal vertebral brucellosis in a 71-year-old mexican-american woman who contracted infection from unpasteurized goat milk. standard agglutination serology was negative; the diagnosis was established by the isolation of brucella melitensis from abscess fluid. a b. melitensis protein microarray comprised of nearly all proteins encoded by the bacterial genome was used to determine the kinetics of this patient's antibody res ...201222219303
unusual presentation of neurobrucellosis: a solitary intracranial mass lesion mimicking a cerebral tumor : a case of encephalitis caused by brucella melitensis.among the diverse presentations of neurobrucellosis, solitary intracranial mass lesions are extremely rare. to the best of our knowledge, we describe here the second case of neurobrucellosis mimicking a cerebral tumor caused by brucella melitensis. the mass lesion was clinically and radiologically indistinguishable from a brain tumor. the diagnosis was established by isolating brucella melitensis in a blood culture and a positive wright's agglutination test on the cerebrospinal fluid at 1:320 ti ...201222231602
lymphocyte activation test for diagnosis of seronegative brucellosis in humans.background: seronegative brucellosis is occasionally encountered in clinical practice especially in localized disease where diagnosis is reached mainly through positive blood culture. cellular immune responses are pivotal for protection against intracellular bacteria such as brucella. this study was performed to evaluate the expression of activation markers on peripheral blood mononuclear cells in response to in vitro stimulation by whole-cell suspension of brucella melitensis for the diagnosis ...201122234109
a large exposure to brucella melitensis in a diagnostic laboratory.background: brucella species are easily transmitted by aerosols and can be acquired in the laboratory. aim: to report the management of a large exposure to brucella melitensis that occurred over six days in a hospital diagnostic laboratory. methods: fifty-one exposed staff were managed according to centers for disease control and prevention guidelines. a further 96 non-exposed laboratory staff were tested for seroprevalence. testing was carried out using the brucella sp. serum agglutination test ...201222237130
characterization of outer membrane vesicles from brucella melitensis and protection induced in mice.the outer membrane vesicles (omvs) from smooth b. melitensis 16 m and a derived rough mutant, vtrm1 strain, were purified and characterized with respect to protein content and induction of immune responses in mice. proteomic analysis showed 29 proteins present in omvs from b. melitensis 16 m; some of them are well-known brucella immunogens such as sod, groes, omp31, omp25, omp19, bp26, and omp16. omvs from a rough vtrm1 induced significantly higher expression of il-12, tnfα, and ifnγ genes in bo ...201122242036
cross-border molecular tracing of brucellosis in europe.to assess the general impact of endemic countries on the re-emergence of brucellosis in non-endemic regions of the european union, the genetic fingerprints of brucella melitensis strains imported to germany were compared to ovine strains from turkey in a molecular epidemiological study. genotyping of 66 brucella strains (based on multiple locus of variable number of tandem repeats analysis) isolated from german travellers and turkish immigrants living in germany revealed epidemiological concorda ...201222277829
efficacy evaluation of live escherichia coli expression brucella p39 protein combined with cpg oligodeoxynucleotides vaccine against brucella melitensis 16m, in balb/c mice.brucella is gram-negative bacteria responsible for brucellosis in a wide variety of animals and humans. balb/c mice were immunized with live escherichia coli expression the p39 gene of brucella melitensis, a gene coding for the periplasmic binding protein. mice were injected with either e. coli bl21 (de3) pet15b or e. coli bl21 (de3) pet15b-p39 alone or adjuvanted with either cpg oligodeoxynucleotides (cpg odn) or non-cpg odn. e. coli bl21 (de3) pet15b-p39 with cpg odn or with non-cpg odn mice g ...201222296786
deep sequencing-based expression transcriptional profiling changes during brucella infection.brucellosis is a worldwide zoonotic infectious disease that has significant economic effects on animal production and human health. the host macrophage -brucella interaction is critical to the establishment of infections. thus, the kinetic transcriptional profile of gene expression in macrophages infected with the brucella melitensis strain 16m was investigated in the current study using a technology based on deep sequencing. the total rna was extracted from macrophages 0, 4, and 24 h post-infec ...201222342430
vaccination programs for reproductive disorders of small ruminants.vaccines are available for the control of contagious epididymitis and abortion in small ruminants, although many of them have significant limitations either in efficacy or safety to both the animals vaccinated and to the people handling the vaccine or animals. shelf-life of vaccines should be extended and improved, so that the vaccine remains effective with longer term storage and ideally without refrigeration, so that use in under-developed rural areas is not restricted (e.g., brucella melitens ...201222364910
autophagy favors brucella melitensis survival in infected macrophages.this study investigated the role of autophagy in the survival of the invasive bacterium brucella melitensis strain 16m in murine macrophages. here, brucella melitensis 16m was found to trigger autophagosome formation, enhance autophagy flux and increase the expression level of the autophagy marker protein lc3-ii. when autophagy was pharmacologically inhibited by 3-methyladenine (3-ma), brucella replication efficiency was significantly decreased (p < 0.05). these results suggest that autophagy fa ...201222367856
brucellosis due to brucella suis in a swine herd associated with a human clinical case in the state of são paulo, brazil.brucella suis has been recognized as the major etiological agent of human brucellosis in areas free from brucella melitensis infection. however, with changes in swine management, the occurrence of swine brucellosis has decreased as has the human incidence of b. suis infection. a swine brucellosis outbreak within a herd from jaboticabal (são paulo, brazil) was detected in july 2006. the herd comprised approximately 300 sows and 1,500 finishing animals. many sows within this herd experienced abort ...201222388711
differential expression of iron acquisition genes by brucella melitensis and brucella canis during macrophage infection.brucella spp. cause chronic zoonotic disease often affecting individuals and animals in impoverished economic or public health conditions; however, these bacteria do not have obvious virulence factors. restriction of iron availability to pathogens is an effective strategy of host defense. for brucellae, virulence depends on the ability to survive and replicate within the host cell where iron is an essential nutrient for the growth and survival of both mammalian and bacterial cells. iron is a par ...201222403618
16s rrna gene sequence analysis of a brucella melitensis infection misidentified as bergeyella zoohelcum.misidentification of brucella species from clinical specimens using commercial bacterial identification systems is a recurring problem. an isolate from a bacterimic patient was identified as bergeyella zoohelcum by microscan walk-away (siemens healthcare diagnostics inc., west sacramento, ca, usa) and as brucella melitensis by vitek 2 system (biomérieux inc., durham, nc, usa). because of this identification ambiguity by the two automated bacterial identification systems we performed 16s rrna seq ...201222421611
the first international standard anti-brucella melitensis serum.the world organisation for animal health (oie) requested an international standard anti-brucella melitensis serum (isabms) to standardise diagnostic tests and reagents for sheep and goats. the agreed criteria were the highest dilution (in negative serum) of the standard which must give a positive result and the lowest dilution (in negative serum) which must simultaneously give a negative result. the two dilutions for each assay were, respectively: indirect enzyme-linked immunosorbent assay (ieli ...201122435193
outbreak investigation of brucellosis in thassos, greece, 2008.in spring 2008, the hellenic center for disease control and prevention was notified about human brucellosis cases in thassos, a greek island that had been up to that point under a brucellosis eradication programme. following the verification of the outbreak a 1:1 case– control study was conducted in the island. the study revealed that consumption of locally produced raw cheese was a risk factor for brucella melitensis infection (odds ratio (or): 15.1, 95% confidence interval (ci): 6.56–34.7). br ...201222449868
characterization of periplasmic protein bp26 epitopes of brucella melitensis reacting with murine monoclonal and sheep antibodies.more than 35,000 new cases of human brucellosis were reported in 2010 by the chinese center for disease control and prevention. an attenuated b. melitensis vaccine m5-90 is currently used for vaccination of sheep and goats in china. in the study, a periplasmic protein bp26 from m5-90 was characterized for its epitope reactivity with mouse monoclonal and sheep antibodies. a total of 29 monoclonal antibodies (mabs) against recombinant bp26 (rbp26) were produced, which were tested for reactivity wi ...201222457830
in situ microscopy analysis reveals local innate immune response developed around brucella infected cells in resistant and susceptible mice.brucella are facultative intracellular bacteria that chronically infect humans and animals causing brucellosis. brucella are able to invade and replicate in a broad range of cell lines in vitro, however the cells supporting bacterial growth in vivo are largely unknown. in order to identify these, we used a brucella melitensis strain stably expressing mcherry fluorescent protein to determine the phenotype of infected cells in spleen and liver, two major sites of b. melitensis growth in mice. in b ...201222479178
transcriptome analysis of hela cells response to brucella melitensis infection: a molecular approach to understand the role of the mucosal epithelium in the onset of the brucella pathogenesis.brucella spp. infect hosts primarily by adhering and penetrating mucosal surfaces, however the initial molecular phenomena of this host:pathogen interaction remain poorly understood. we hypothesized that characterizing the epithelial-like human hela cell line molecular response to wild type brucella melitensis infection would help to understand the role of the mucosal epithelium at the onset of the brucella pathogenesis. rna samples from b. melitensis-infected hela cells were taken at 4 and 12 h ...201222484383
otpr regulated the growth, cell morphology of b. melitensis and tolerance to β-lactam agents.the intracellular pathogen, brucella melitensis, possesses an operon with two components: otpr (bmei0066), which encodes a response regulator, and bmei0067, which encodes a putative camp-dependent protein kinase regulatory subunit. previous studies have shown that a polar mutation in the bmei0066 gene significantly decreased virulence and stress tolerance in brucella. in this study, we constructed non-polar mutant with deletion of otpr, as well as its complementary strain to further investigate ...201222503392
evaluation of sample preparation methods for maldi-tof ms identification of highly dangerous bacteria.to propose a universal workflow of sample preparation method for the identification of highly pathogenic bacteria by maldi-tof ms.201222512320
human to human transmission of brucella melitensis.human brucellosis is acquired mainly through contact with infected animal tissues, ingestion of unpasteurized dairy products or infected aerosols. person to person transmission is still considered uncertain. the case of a woman diagnosed with proven brucellosis after her husband suffered a relapse of bacteremia with brucella melitensis biotype 3, which was originally acquired abroad by eating goat cheese, is described. it was postulated that person to person spread of brucellosis is a likely mod ...199522514390
protective role of antibodies induced by brucella melitensis b115 against b. melitensis and brucella abortus infections in mice.it has been demonstrated that antibodies specific for o-ps antigen of brucella smooth strains are involved in the protective immunity of brucellosis. since the rough strain brucella melitensis b115 was able to protect mice against wild brucella strains brucellosis despite the lack of anti-ops antibodies, in this study we evaluated the biological significance of antibodies induced by this strain, directed to antigens other than o-ps, passively tranferred to untreated mice prior to infection with ...201222521283
purification, refolding and characterization of the trimeric omp2a outer membrane porin from brucella melitensis.brucella melitensis is a gram-negative bacteria known to cause brucellosis and to produce severe infections in humans. whilst brucella's outer membrane proteins have been extensively studied due to their potential role as antigens or virulence factors, their function is still poorly understood at the structural level, as the 3d structure of brucella β-barrel membrane proteins are still unknown. in this context, the b. melitensis trimeric omp2a porin has been overexpressed and refolded in n-dodec ...201222538317
optimization and efficient purification of recombinant omp28 protein of brucella melitensis using triton x-100 and β-mercaptoethanol.the high level expression of recombinant proteins in escherichia coli often leads to the formation of inclusion bodies that contain most of the expressed protein held together by non-covalent forces. the inclusion bodies are usually solubilized using strong denaturing agents like urea and guanidium hydrochloride. in this study recombinant omp28 (romp28) protein of brucella melitensis was expressed in two different vector systems and further efficient purification of the protein was done by modif ...201222542588
invasive escherichia coli vaccines expressing brucella melitensis outer membrane proteins 31 or 16 or periplasmic protein bp26 confer protection in mice challenged with b. melitensis.because of the serious economic and medical consequences of brucellosis, efforts are to prevent infection of domestic animals through vaccines. many disadvantages are associated with the current brucella melitensis rev.1 vaccine prompting development of alternative vaccines and delivery. escherichia coli (dh5α) was engineered to express a plasmid containing the inv gene from yersinia pseudotuberculosis and the hly gene from listeria monocytogenes. these recombinant invasive e. coli expressing b. ...201222546330
active evasion of ctl mediated killing and low quality responding cd8+ t cells contribute to persistence of brucellosis.brucellosis is a common zoonotic disease that remains endemic in many parts of the world. dissecting the host immune response during this disease provides insight as to why brucellosis is often difficult to resolve. we used a brucella epitope specific in vivo killing assay to investigate the ability of cd8+ t cells to kill targets treated with purified pathogenic protein. importantly, we found the pathogenic protein tcpb to be a novel effector of adaptive immune evasion by inhibiting cd8+ t cell ...201222558103
predicting antigenicity of proteins in a bacterial proteome; a protein microarray and naïve bayes classification approach.discovery of novel antigens associated with infectious diseases is fundamental to the development of serodiagnostic tests and protein subunit vaccines against existing and emerging pathogens. efforts to predict antigenicity have relied on a few computational algorithms predicting signal peptide sequences (signalp), transmembrane domains, or subcellular localization (psort). an empirical protein microarray approach was developed to scan the entire proteome of any infectious microorganism and empi ...201222589097
misidentification of brucella melitensis as bergeyella zoohelcum by microscan walkaway®: a case report.to describe the misidentification of brucella melitensis as bergeyella zoohelcum by microscan walkaway®, a commonly used bacterial identification system.201222614245
ifn-γ-deficient mice develop il-1-dependent cutaneous and musculoskeletal inflammation during experimental brucellosis.human brucellosis exhibits diverse pathological manifestations that can affect almost any organ. in particular, osteoarticular complications are the most common focal manifestation of brucellosis and occur in 40-80% of patients. in immunocompetent mice, brucella replication is generally restricted to the spleen, liver, and to a lesser extent, lns, thereby limiting their use for study of focal inflammation often found in brucellosis. here, we report that nasal, oral, or peritoneal infection of if ...201222636321
molecular cloning of virb12 gene of brucella melitensis 16m strain in pet28a vector.to clone the virb12 gene in pet28a expression vector for production of recombinant protein to be used as antigenic component for future serological test development.201222647810
extrapulmonary tuberculosis: importance of differential diagnosis.patients with tuberculosis (tb) typically present with pulmonary tb, but extrapulmonary tb is also an important clinical problem. in particular, osteoarticular tb has a great morbidity and its diagnosis gold-standard includes culture and histopathology of the infected area. the present case study describes a 53-year-old female portuguese farmer who had initial clinical and imaging features suggestive of tuberculous osteomyelitis. however, she denied any tb contacts and the tuberculin skin-test a ...201122679255
isolation of brucella melitensis biotype 3 from epidural empyema in a bosnian immigrant in germany.brucellosis is a regionally emerging infectious disease in mediterranean countries with an increasing number of human cases and high morbidity rates. here, we describe a case of severe b. melitensis biotype 3 infection in an immigrant who had contact with ruminants during a short-term stay in bosnia before he returned to germany. the patient developed thoracic spondylodiscitis accompanied by a large epidural empyema and neurological deficits. the isolated strain was characterized and compared to ...201222683655
brucella arthritis of the knee, 1 year after revision of anterior cruciate ligament reconstruction.brucellosis is a zoonotic infection with a broad spectrum of clinical manifestations. the authors report the first case in the literature of septic arthritis of the knee 1 year after revision of anterior cruciate ligament reconstruction. brucella melitensis biotype 3 was found in both synovial fluid and blood cultures. the patient was treated initially with arthroscopic debridement. after the diagnosis was confirmed, a second arthroscopic lavage and metal work removal was applied leaving the gra ...201122700607
brucellosis presenting as septic shock.brucellosis generally presents with fever, malaise, weight loss and bone pain with either an abrupt or insidious onset. a 76-year-old man presented in april 2010 with fever of 103°f, severe tachycardia, tachypnoea and a blood pressure of 80/50 mm hg requiring fluids and vasopressor support with norepinephrine. the patient had brucellosis in 1956 which was treated for 'many weeks' with tetracycline and streptomycin. he has had no recurrences since that time. he denies recent travel outside the us ...201122701076
[spondylodiscitis: which etiology?].spondylodiskitis is a rare condition in adults but with high index of morbidity and mortality due to difficult diagnosis. the insidious evolution and the unspecificity of symptoms interfere with prompt treatment, being the diagnosis based primarily on a high index of suspicion. there are two major groups of spondylodiskitis: pyogenic, being staphylococcus aureus the most frequent etiological pathogen, and granulomatous, caused mainly by mycobacterium tuberculosis and brucella melitensis, both en ...201122713202
induction of protective immunity against brucellosis in mice by vaccination with a combination of naloxone, alum, and heat-killed brucella melitensis 16 m.a t-helper cell type 1-specific response leads to the elimination of intracellular infection with brucella. studies have shown that naloxone (nlx) can promote a cellular immune response in this respect. the current study was carried out to evaluate the induction of protective immunity in mice against brucellosis by vaccination with a combination of nlx, alum, and heat-killed brucella melitensis 16 m (hkb).201322727892
inhibition of mcd14 inhibits tnfα secretion and no production in raw264.7 cells stimulated by brucella melitensis infection.brucellosis, caused by brucella spp., is an important disease affecting not only human health, but also a number of animal species around the world. a receptor for lps of brucella and important innate immune molecule, cd14, has been implicated in the initiation of the inflammatory response to sepsis. evidence indicates that upstream inhibition of the lps initiated inflammatory pathway is an effective therapeutic approach for attenuating damaging immune activation. the aim of this study was to ex ...201222770519
the pathophysiology of inhalational brucellosis in balb/c mice.to characterize the clinical presentation and pathophysiology of inhalational brucellosis, balb/c mice were challenged with brucella melitensis 16m in a nose-only aerosol exposure chamber. a low dose of 1000 cfu/animal of b. melitensis resulted in 45% of mice with tissue burdens eight weeks post-challenge. the natural history of brucellosis in mice challenged by higher aerosol doses was examined by serial euthanizing mice over an eight week period. higher challenge doses of 1.00e+05 and 5.00e+05 ...201222773944
evaluation of the vitek 2 gram-negative (gn) microbial identification test card: collaborative study.a collaborative study was conducted to evaluate the performance of the vitek 2 gram-negative (gn) identification card for use with the vitek 2 automated microbial identification system. the gn test card is used in the identification of fermenting and nonfermenting gram-negative bacilli, including the select agent organisms brucella melitensis, francisella tularensis, burkholderia mallei, b. pseudomallei, and yersinia pestis. the vitek 2 gn card is based on 47 biochemical tests measuring carbon s ...201222816270
the two-component system prls/prlr of brucella melitensis is required for persistence in mice and appears to respond to ionic strength.bacterial adaptation to environmental conditions is essential to ensure maximal fitness in the face of several stresses. in this context, two-component systems (tcss) represent a predominant signal transduction mechanism, allowing an appropriate response to be mounted when a stimulus is sensed. as facultative intracellular pathogens, brucella spp. face various environmental conditions, and an adequate response is required for a successful infection process. recently, bioinformatic analysis of br ...201222859617
withdrawn: incidence study of brucella abortus and brucella melitensis in bovine and buffalo semen samples by real-time pcr assay in iran.ahead of print article withdrawn by publisher.201222864410
an uncommon case of pediatric neurobrucellosis associated with intracranial hypertension.we present the case of a 4-year-old boy who was admitted to hospital with intracranial hypertension, headache, diplopia, papilledema, and a normal brain mri. brucella melitensis in the cerebrospinal fluid was confirmed with pcr assay. we believe that neurobrucellosis should be included in the differential diagnosis when headaches persist following brucellosis. in addition, we suggest that when cerebrospinal fluid culture is negative, pcr may prove to be an optimal alternative tool for an immedia ...201222900217
microrna expression profile in raw264.7 cells in response to brucella melitensis infection.microrna (mirna) is small non-coding rna with approximate 22 nt in length. recent studies indicate that mirnas play significant roles in pathogen-host interactions. brucella organisms are gram-negative facultative intracellular bacteria that cause brucellosis. brucella strains infect macrophages and establish chronic infection by altering host life activities including apoptosis and autophagy. here, we report a comprehensive analysis of mirna expression profiles in mock- and brucella-infected ra ...201222904669
a rare case of seronegative culture--proven infection with brucella suis.brucellosis is a chronic infection produced by members of the brucella family. diagnosis of this condition requires either isolation of the organism in culture or positive serological tests.we describe a 27-year-old male admitted as a case of pyrexia of unknown origin (puo), who tested negative for brucella igm elisa test on preliminary evaluation but was subsequently diagnosed on the strength of positive blood and bone marrow cultures to be a case of brucellosis secondary to brucella suis infec ...201222905059
mlva16 typing of portuguese human and animal brucella melitensis and brucella abortus isolates.to investigate the epidemiological relationship of isolates from different portuguese geographical regions and to assess the diversity among isolates, the mlva16(orsay) assay (panels 1, 2a and 2b) was performed with a collection of 126 brucella melitensis (46 human and 80 animal isolates) and 157 b. abortus field isolates, seven vaccine strains and the representative reference strains of each species. the mlva16(orsay) showed a similar high discriminatory power (hgdi 0.972 and 0.902) for both sp ...201222905141
surgical management of cervical spinal epidural abscess caused by brucella melitensis : report of two cases and review of the literature.spinal epidural abscess, if especially caused by brucellosis is a very rare disease which is usually a consequence of spondylodiscitis. the spinal column can be affected at any joint; however, the lumbar spine is the most common region, especially at the level of the l4-5 and l5-s1. the frequency of spinal involvement usually seen at the lumbar, thoracic and cervical spine respectively. as an occupational disease in farmers, veterinaries, butchers, laboratory staff and shepherds, brucellosis can ...201222949972
genome sequence of brucella melitensis s66, an isolate of sequence type 8, prevalent in china.brucella melitensis is the most-represented brucella species causing human brucellosis in china. here we report the complete genome sequence of b. melitensis strain s66, a representative strain of sequence type 8 (st8), which is prevalent in china, making it possible to compare the genome sequences of isolates from different countries.201222965081
genome sequences of brucella melitensis 16m and its two derivatives 16m1w and 16m13w, which evolved in vivo.brucella melitensis is an intracellular pathogen that induces chronic infection in humans. here, we report the genome sequences of 16m and its two derivatives, 16m1w and 16m13w, which were allowed to adapt in vivo for 1 and 13 weeks, respectively. our findings contribute to the investigation of adaptive mutations and mechanisms of chronic infection by b. melitensis.201222965104
human brucellosis outbreak acquired through camel milk ingestion in southern israel.human brucellosis is common in southern israel among the semi-nomadic bedouin, a population that consumes unpasteurized dairy products. though camel milk ingestion is a known mechanism for brucellosis acquisition, only a few reports of sporadic cases have been published in the medical literature.201222977965
[brucella suis--a little-known zoonotic agent].brucellosis is a rare but serious bacterial zoonosis. officially, the czech republic is among countries that are free from brucellosis in livestock. in the country, sporadic imported human infections may occur, caused by e.g. brucella melitensis. in wild hare populations, however, rare cases of infection caused by brucella suis are still observed, potentially threatening humans. the short communication reports two cases of hare brucellosis in moravia, czech republic, and experiences with isolate ...201222997779
crucial role of gamma interferon-producing cd4+ th1 cells but dispensable function of cd8+ t cell, b cell, th2, and th17 responses in the control of brucella melitensis infection in mice.brucella spp. are facultative intracellular bacterial pathogens responsible for brucellosis, a worldwide zoonosis that causes abortion in domestic animals and chronic febrile disease associated with serious complications in humans. there is currently no approved vaccine against human brucellosis, and antibiotic therapy is long and costly. development of a safe protective vaccine requires a better understanding of the roles played by components of adaptive immunity in the control of brucella infe ...201223006848
evaluation of protective effect of dna vaccines encoding the bab1_0263 and bab1_0278 open reading frames of brucella abortus in balb/c mice.the immunogenicity of two dna vaccines encoding open reading frames (orfs) of genomic island 3 (gi-3), specific for brucella abortus and brucella melitensis, has been examined. intramuscular injection of plasmid dna carrying the bab1_0263 and bab1_0278 genes (pvf263 and pvf278, respectively) into balb/c mice elicited both humoral and cellular immune responses. mice injected with pvf263 or pvf278 had a dominant immunoglobulin g2a (igg2a) response. in addition, both dna vaccines elicited a t-cell- ...201223026687
genome sequences of three live attenuated vaccine strains of brucella species and implications for pathogenesis and differential diagnosis.live attenuated vaccines play essential roles in the prevention of brucellosis. here, we report the draft genome sequences of three vaccine strains, brucella melitensis m5-10, b. suis s2-30, and b. abortus 104m. primary genome sequence analysis identified mutations, deletions, and insertions which have implications for attenuation and signatures for differential diagnosis.201223045513
silencing of vamp3 expression does not affect brucella melitensis infection in mouse macrophages.it has been proposed that intracellular pathogens may interfere with expression or function of proteins that mediate vesicular traffic in order to survive inside cells. brucella melitensis is an intracellular pathogen that evades phagosome-lysosome fusion, surviving in the so-called brucella-containing vacuoles (bcv). vesicle-associated membrane protein 3 (vamp3) is a v-snare protein that promotes the exocytosis of the proinflammatory cytokine tnf at the phagocytic cup when docking to its cognat ...201223076244
exact alpha-error determination for two-stage sampling strategies to substantiate freedom from disease.sampling strategies to substantiate freedom from disease are important when it comes to the trade of animals and animal products. when considering imperfect tests and finite populations, sample size calculation can, however, be a challenging task. the generalized hypergeometric formula developed by cameron and baldock (1998a) offers a framework that can elegantly be extended to multi-stage sampling strategies, which are widely used to account for disease clustering at herd-level. the achieved al ...201323103193
complete genome sequence of brucella melitensis biovar 3 strain ni, isolated from an aborted bovine fetus.from an aborted bovine fetus in china, a bacterial strain named ni was isolated and identified as brucella melitensis by a pcr assay. strain ni was further characterized as b. melitensis biovar 3 using biochemical assays. here we report the complete genome sequence of strain ni.201223105063
multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness.the centers for disease control and prevention and united states army research institute for infectious diseases have developed real-time pcr assays for the detection of bioterrorism threat agents. these assays all rely on a limited number of approved real-time pcr master mixes. because the availability of these reagents is a critical element of bioterrorism preparedness, we undertook a joint national preparedness exercise to address the potential surge needs resulting from a large-scale bio-eme ...201223107058
an atypical presentation of brucellosis in a patient with isolated thrombocytopenia complicated with upper gastrointestinal tract bleeding.a 59-year-old female patient was admitted to the emergency service with complaints of hematemesis and melena for the last few days. in laboratory tests, the platelet count was found to be 6 × 10(9)/l. intravenous or oral corticosteroid treatment was thought to be given for itp but disclaimed due to upper gis bleeding. on the 5th day of treatment, brucella melitensis was isolated from blood culture before the results of wright tube agglutination tests were reported positive as 1 : 80. on the seco ...201223118764
retraction: incidence study of brucella abortus and brucella melitensis in bovine and buffalo semen samples by real-time pcr assay in iran.this article released online on august 3, 2012 as advance publication was withdrawn from consideration for publication in the journal of veterinary medical science at author's request.201223123884
brucella melitensis mucr, an orthologue of sinorhizobium meliloti mucr, is involved in resistance to oxidative, detergent, and saline stresses and cell envelope modifications.brucella spp. and sinorhizobium meliloti are alphaproteobacteria that share not only an intracellular lifestyle in their respective hosts, but also a crucial requirement for cell envelope components and their timely regulation for a successful infectious cycle. here, we report the characterization of brucella melitensis mucr, which encodes a zinc finger transcriptional regulator that has previously been shown to be involved in cellular and mouse infections at early time points. mucr modulates th ...201323161025
pluronic p85 enhances the efficacy of outer membrane vesicles as a subunit vaccine against brucella melitensis challenge in mice.brucellosis is the most common zoonotic disease worldwide, and there is no vaccine for human use. brucella melitensis rev1, a live attenuated strain, is the commercial vaccine for small ruminants to prevent b. melitensis infections but has been associated with abortions in animals. moreover, strain rev1 is known to cause disease in humans and cannot be used for human vaccination. outer membrane vesicles (omvs) obtained from b. melitensis have been shown to provide protection similar to strain re ...201223163875
fed batch fermentation and purification strategy for high yield production of brucella melitensis recombinant omp 28 kda protein and its application in disease diagnosis.brucellosis is a disease caused by bacteria belonging to the genus brucella. it affects cattle, goat, sheep, dog and humans. the serodiagnosis of brucellosis involves detection of antibodies generated against the lps or whole cell bacterial extracts, however these tests lack sensitivity and specificity. the present study was performed to optimize the culture condition for the production of recombinant brucella melitensis outer membrane protein 28 kda protein in e.coli via fed batch fermentation. ...201323167982
isolation and molecular characterization of brucella melitensis from seropositive goats in peninsula malaysia.a study was carried out to isolate brucella melitensis using established bacteriological and pcr techniques in brucella seropositive goats in farms in selangor, negeri sembilan, melaka and pulau pinang. brucella melitensis was isolated from 7 of 134 reactors with the highest isolation from the vaginal swabs (57.14%) followed by the spleen (28.57%), uterine fluid (14.29%). no brucella was isolated from the lymph nodes. pcr confirmed all the seven isolates as b. melitensis and isolates were phylog ...201223202595
complete genome sequence of brucella melitensis 133, an isolate of biovar 1 of sequence type 32.brucellosis is highly epidemic in china. of the six classical species, brucella melitensis and biovar 1 are the most represented species and biovar that cause human brucellosis in china. here, we report the genome sequence of brucella melitensis strain 133, a strain of biovar 1 of sequence type 32.201223209199
genome sequence of brucella melitensis strain 128, an isolate of biovar 3 of sequence type 8.brucella melitensis is the most common brucella species causing human brucellosis. b. melitensis is divided into 3 biovars. here, we report the complete genome sequence of b. melitensis strain 128, a strain of biovar 3 of sequence type 8, which is prevalent in china.201223209219
sustained and differential antibody responses to virulence proteins of brucella melitensis during acute and chronic infections in human brucellosis.brucellosis is an important zoonotic disease caused primarily by the bacterial pathogens brucella melitensis and b. abortus. the pathogens cause debilitating febrile illness that can progress into a long-lasting disease with severe complications in humans. understanding the mechanisms by which the host immune system responds to the infection will provide important information on the pathogenesis and development of differential diagnostic assays. in this study, a protein microarray was used to ev ...201323224716
innate immune recognition of flagellin limits systemic persistence of brucella.brucella are facultative intracellular bacteria that cause chronic infections by limiting innate immune recognition. it is currently unknown whether brucella flic flagellin, the monomeric subunit of flagellar filament, is sensed by the host during infection. here, we used two mutants of brucella melitensis, either lacking or overexpressing flagellin, to show that flic hinders bacterial replication in vivo. the use of cells and mice genetically deficient for different components of inflammasomes ...201323227931
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