Publications

the polymeric antigen blsomp31 confers protection against brucella ovis infection in rams.	we have engineered the polymeric vaccine blsomp31 by decorating the highly immunogenic and decameric brucella lumazine synthase with an exposed loop of the brucella outer membrane protein omp31. in the present study, we have immunized different groups of rams with the recombinant chimera rblsomp31 in two different adjuvants (incomplete freund adjuvant-ifa and quil a) and with the plasmid pciblsomp31 administered either by i.m. injection alone or by using electroporation. in addition, we have use ...	2009	19748579
mutation of his465 alters the ph-dependent spectroscopic properties of escherichia coli glutamate decarboxylase and broadens the range of its activity toward more alkaline ph.	glutamate decarboxylase (gadb) from escherichia coli is a hexameric, pyridoxal 5'-phosphate-dependent enzyme catalyzing co(2) release from the alpha-carboxyl group of l-glutamate to yield gamma-aminobutyrate. gadb exhibits an acidic ph optimum and undergoes a spectroscopically detectable and strongly cooperative ph-dependent conformational change involving at least six protons. crystallographic studies showed that at mildly alkaline ph gadb is inactive because all active sites are locked by the ...	2009	19797049
survival of the fittest: how brucella strains adapt to their intracellular niche in the host.	brucella strains produce abortion and infertility in their natural hosts and a zoonotic disease in humans known as undulant fever. these bacteria do not produce classical virulence factors, and their capacity to successfully survive and replicate within a variety of host cells underlies their pathogenicity. extensive replication of the brucellae in placental trophoblasts is associated with reproductive tract pathology in natural hosts, and prolonged persistence in macrophages leads to the chroni ...	2009	19830453
brucellosis: the case for live, attenuated vaccines.	the successful control of animal brucellosis and associated reduction in human exposure has limited the development of human brucellosis vaccines. however, the potential use of brucella in bioterrorism or biowarfare suggests that direct intervention strategies are warranted. although the dominant approach has explored the use of live attenuated vaccines, side effects associated with their use has prevented widespread use in humans. development of live, attenuated brucella vaccines that are safe ...	2009	19837284
evaluation of automated and manual commercial dna extraction methods for recovery of brucella dna from suspensions and spiked swabs.	this study evaluated automated and manual commercial dna extraction methods for their ability to recover dna from brucella species in phosphate-buffered saline (pbs) suspension and from spiked swab specimens. six extraction methods, representing several of the methodologies which are commercially available for dna extraction, as well as representing various throughput capacities, were evaluated: the magna pure compact and the magna pure lc instruments, the it 1-2-3 dna sample purification kit, t ...	2009	19846627
evaluation of three different antigens in an indirect enzyme-linked immunoassay for the detection of antibodies against brucella abortus srb51 in vaccinated heifers.	the live attenuated brucella abortus srb51 (srb51) is a partial o-chain-deprived mutant. the relative lack of the polysaccharide prevents it from inducing antibodies detectable by most of the serological tests used for the diagnosis of bovine brucellosis. the performance of three antigens used in an indirect enzyme-linked immunoassay test for detecting srb51 antibodies were evaluated. a homogeneous group of twenty-five 10-month-old hereford heifers was used. the animals were bled on day 0 and th ...	2009	18980780
immunization with recombinant brucella species outer membrane protein omp16 or omp19 in adjuvant induces specific cd4+ and cd8+ t cells as well as systemic and oral protection against brucella abortus infection.	available vaccines against brucella spp. are live attenuated brucella strains. in order to engineer a better vaccine to be used in animals and humans, our laboratory aims to develop an innocuous subunit vaccine. particularly, we are interested in the outer membrane proteins (omps) of b. abortus: omp16 and omp19. in this study, we assessed the use of these proteins as vaccines against brucella in balb/c mice. immunization with lipidated omp16 (l-omp16) or l-omp19 in incomplete freund's adjuvant ( ...	2009	18981242
immunization with recombinant brucella species outer membrane protein omp16 or omp19 in adjuvant induces specific cd4+ and cd8+ t cells as well as systemic and oral protection against brucella abortus infection.	available vaccines against brucella spp. are live attenuated brucella strains. in order to engineer a better vaccine to be used in animals and humans, our laboratory aims to develop an innocuous subunit vaccine. particularly, we are interested in the outer membrane proteins (omps) of b. abortus: omp16 and omp19. in this study, we assessed the use of these proteins as vaccines against brucella in balb/c mice. immunization with lipidated omp16 (l-omp16) or l-omp19 in incomplete freund's adjuvant ( ...	2009	18981242
efficacy of bp26 and bp26/omp31 b. melitensis rev.1 deletion mutants against brucella ovis in rams.	brucella melitensis rev.1 is the most effective vaccine against b. ovis infection in sheep but induces antibodies interfering with b. melitensis diagnosis. brucella bp26 and omp31 proteins are differential diagnostic antigens. single or double bp26 and omp31 rev.1 deletion mutants have been proven effective against b. melitensis in sheep. here, the cgv26 (deleted in bp26 gene) and cgv2631 (deleted in both bp26 and omp31 genes) mutants have been tested for efficacy against b. ovis in rams. either ...	2009	19007836
differential expression of inflammatory and immune response genes in rams experimentally infected with a rough virulent strain of brucella ovis.	infection of sheep with brucella ovis results in ovine brucellosis, a disease characterized by infertility in rams, abortion in ewes and increased perinatal mortality in lambs. during the course of the infection both the ovine immune response and host cell gene expression are modified. the objective of this research was to conduct a preliminary characterization of differential gene expression in rams experimentally infected with b. ovis by microarray hybridization and real-time rt-pcr. of the 60 ...	2009	19056128
improved prediction of malaria degradomes by supervised learning with svm and profile kernel.	the spread of drug resistance through malaria parasite populations calls for the development of new therapeutic strategies. however, the seemingly promising genomics-driven target identification paradigm is hampered by the weak annotation coverage. to identify potentially important yet uncharacterized proteins, we apply support vector machines using profile kernels, a supervised discriminative machine learning technique for remote homology detection, as a complement to the traditional alignment ...	2009	19057851
improved prediction of malaria degradomes by supervised learning with svm and profile kernel.	the spread of drug resistance through malaria parasite populations calls for the development of new therapeutic strategies. however, the seemingly promising genomics-driven target identification paradigm is hampered by the weak annotation coverage. to identify potentially important yet uncharacterized proteins, we apply support vector machines using profile kernels, a supervised discriminative machine learning technique for remote homology detection, as a complement to the traditional alignment ...	2009	19057851
a flood of microbial genomes-do we need more?		2009	19513110
brucella melitensis, b. neotomae and b. ovis elicit common and distinctive macrophage defense transcriptional responses.	brucella spp. establish an intracellular replicative niche in macrophages, while macrophages attempt to eliminate the bacteria by innate defense mechanisms. brucella spp. possess similar genomes yet exhibit different macrophage infections. few b. melitensis and b. neotomae enter macrophages with intracellular adaptation occurring over 4-8 hr. conversely, b. ovis are readily ingested by macrophages and exhibit a persistent plateau of infection. evaluating early macrophage interaction with brucell ...	2009	19934366
the consequences of genetic drift for bacterial genome complexity.	genetic drift, which is particularly effective within small populations, can shape the size and complexity of genomes by affecting the fixation of deleterious mutations. in bacteria, assessing the contribution of genetic drift to genome evolution is problematic because the usual methods, based on intraspecific polymorphisms, can be thwarted by difficulties in delineating species' boundaries. the increased availability of sequenced bacterial genomes allows application of an alternative estimator ...	2009	19502381
tir domain-containing adaptor sarm is a late addition to the ongoing microbe-host dialog.	toll/interleukin-1 receptor (tir) domain-containing proteins play important roles in defense against pathogens in both animals and plants, connecting the immunity signaling pathways via a chain of specific protein-protein interactions. among them is sarm, the only tir domain-containing adaptor that can negatively regulate tlr signaling. by extensive phylogenetic analysis, we show here that sarm is closely related to bacterial proteins with tir domains, suggesting that this family has a different ...	2010	21110998
tir domain-containing adaptor sarm is a late addition to the ongoing microbe-host dialog.	toll/interleukin-1 receptor (tir) domain-containing proteins play important roles in defense against pathogens in both animals and plants, connecting the immunity signaling pathways via a chain of specific protein-protein interactions. among them is sarm, the only tir domain-containing adaptor that can negatively regulate tlr signaling. by extensive phylogenetic analysis, we show here that sarm is closely related to bacterial proteins with tir domains, suggesting that this family has a different ...	2010	21110998
polymer nanoparticles containing tumor lysates as antigen delivery vehicles for dendritic cell-based antitumor immunotherapy.	encapsulation of tumor-associated antigens in polymer nanoparticles (np) is a promising approach to enhance efficiency of antigen delivery for anti-tumor vaccines. head and neck squamous carcinoma (hnscc) cell lines were initially used to generate tumor-associated antigens (taa)-containing poly (lactic-co-glycolic acid) (plga) np; encapsulation efficiency and release kinetics were profiled. findings were adopted to entrap fresh tumor lysate from five patients with advanced hnscc. to test the hyp ...	2010	20692374
polymer nanoparticles containing tumor lysates as antigen delivery vehicles for dendritic cell-based antitumor immunotherapy.	encapsulation of tumor-associated antigens in polymer nanoparticles (np) is a promising approach to enhance efficiency of antigen delivery for anti-tumor vaccines. head and neck squamous carcinoma (hnscc) cell lines were initially used to generate tumor-associated antigens (taa)-containing poly (lactic-co-glycolic acid) (plga) np; encapsulation efficiency and release kinetics were profiled. findings were adopted to entrap fresh tumor lysate from five patients with advanced hnscc. to test the hyp ...	2010	20692374
detection of brucella ovis in ovine from paraíba state, in the northeast region of brazil.	to determine the presence of brucella ovis in ovine from paraíba state, in the northeast region of brazil, 80 animals slaughtered in the public slaughterhouse of patos city were used. before slaughter, blood samples were collected by jugular venopuncture from each animal, and after slaughter, testicles, epidydimus and uterus were aseptically collected. for the serological diagnosis of b. ovis and b. abortus infections, the agar gel immunodiffusion (agid) and rose bengal (rbt) tests were carried ...	2010	24031505
cholesterol, ganglioside gm1 and class a scavenger receptor contribute to infection by brucella ovis and brucella canis in murine macrophages.	the establishment of infection by brucella ovis and brucella canis in j774.a1 macrophages was found to be dependent upon cholesterol and ganglioside gm(1), two components of lipid rafts. this process also required a class a scavenger receptor of macrophages, and was not inhibited by smooth and rough lipopolysaccharides from brucella spp. in response to infection, both bacteria induced a weak degree of macrophage activation. these results demonstrate that b. ovis and b. canis use cell surface rec ...	2010	20083220
atp-binding cassette systems of brucella.	brucellosis is a prevalent zoonotic disease and is endemic in the middle east, south america, and other areas of the world. in this study, complete inventories of putative functional abc systems of five brucella species have been compiled and compared. abc systems of brucella melitensis 16m, brucella abortus 9-941, brucella canis rm6/66, brucella suis 1330, and brucella ovis 63/290 were identified and aligned. high numbers of abc systems, particularly nutrient importers, were found in all brucel ...	2010	20169092
development and evaluation of a species-specific pcr assay for the detection of brucella ovis infection in rams.	brucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. the goal of this study was to develop a pcr test targeting specific b. ovis genomic sequences. specific primer pairs were designed targeting 12 of those orfs. samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams (n=9) every other week up to 180 days post infection (dpi), when tissue sam ...	2010	20347534
pentaplexed quantitative real-time pcr assay for the simultaneous detection and quantification of botulinum neurotoxin-producing clostridia in food and clinical samples.	botulinum neurotoxins are produced by the anaerobic bacterium clostridium botulinum and are divided into seven distinct serotypes (a to g) known to cause botulism in animals and humans. in this study, a multiplexed quantitative real-time pcr assay for the simultaneous detection of the human pathogenic c. botulinum serotypes a, b, e, and f was developed. based on the taqman chemistry, we used five individual primer-probe sets within one pcr, combining both minor groove binder- and locked nucleic ...	2010	20435756
the feasibility of using antigens prepared with rough brucella strains for diagnosis of canine brucellosis.	clinical diagnosis of canine brucellosis is not sensitive enough and a negative blood culture cannot rule out the disease. indirect methods of serological testing such as agar gel immunodiffusion (agid), rapid slide agglutination test (rsat) and indirect enzyme linked immunoassay (ielisa) are preferred for routine diagnosis. since brucella canis shares antigenic components with the brucella ovis and brucella abortus rb51 strain, it would seem that either strain could be used as antigen. we prese ...	2010	20461292
proteomics-based confirmation of protein expression and correction of annotation errors in the brucella abortus genome.	brucellosis is a major bacterial zoonosis affecting domestic livestock and wild mammals, as well as humans around the globe. while conducting proteomics studies to better understand brucella abortus virulence, we consolidated the proteomic data collected and compared it to publically available genomic data.	2010	20462421
brucella taxonomy and evolution.	taxonomy and nomenclature represent man-made systems designed to enhance understanding of the relationship between organisms by comparison of discrete sets of properties. initial efforts at bacterial taxonomy were flawed as a result of the previous use of nonsystematic approaches including common names resulting in confusing and inaccurate nomenclature. a decision was made to start afresh with bacterial nomenclature and to avoid the hazards experienced in the taxonomic classification of higher o ...	2010	20521932
genomic comparisons of brucella spp. and closely related bacteria using base compositional and proteome based methods.	classification of bacteria within the genus brucella has been difficult due in part to considerable genomic homogeneity between the different species and biovars, in spite of clear differences in phenotypes. therefore, many different methods have been used to assess brucella taxonomy. in the current work, we examine 32 sequenced genomes from genus brucella representing the six classical species, as well as more recently described species, using bioinformatical methods. comparisons were made at t ...	2010	20707916
effect of extender supplementation with various antimicrobial agents on viability of brucella ovis and actinobacillus seminis in cryopreserved ovine semen.	the objective was to determine the effectiveness of various antimicrobial agents added to semen extender for inactivation of b. ovis or a. seminis in ovine semen after cryopreservation. in experiment 1, 20 ejaculates from a crossbred ram infected with b. ovis were cryopreserved in tris-based extenders with various antimicrobial agents: (i) control without antibiotics, (ii) with penicillin and streptomycin (1000 iu/ml and 1 mg/ml, respectively), (iii) lincomycin (0.15 mg/ml), (iv) sulphadiazine ( ...	2010	20708246
diagnosis of brucellosis in livestock and wildlife.	to describe and discuss the merits of various direct and indirect methods applied in vitro (mainly on blood or milk) or in vivo (allergic test) for the diagnosis of brucellosis in animals.	2010	20718082
obstacles of multiplex real-time pcr for bacterial 16s rdna: primer specifity and dna decontamination of taq polymerase.	background: the detection of a broad range of bacteria by pcr is applied for the screening of blood and blood products with special attention to platelet concentrates. for practical use it is desirable that detection systems include gram-positive, gram-negative and non-gram-stainable bacteria. it is quite challenging to achieve high sensitivity along with a clear negative control with pcr reagents, because especially taq polymerase is contaminated with traces of bacterial dna. methods: bacterial ...	2010	20737013
genomic island 2 is an unstable genetic element contributing to brucella lipopolysaccharide spontaneous smooth-to-rough dissociation.	brucella is a gram-negative bacterium that causes a worldwide-distributed zoonosis. the genus includes smooth (s) and rough (r) species that differ in the presence or absence, respectively, of the o-polysaccharide of lipopolysaccharide. in s brucellae, the o-polysaccharide is a critical diagnostic antigen and a virulence determinant. however, s brucellae spontaneously dissociate into r forms, a problem in antigen and s vaccine production. spontaneous r mutants of brucella abortus, brucella melit ...	2010	20952568
biological warfare agents.	the recent bioterrorist attacks using anthrax spores have emphasized the need to detect and decontaminate critical facilities in the shortest possible time. there has been a remarkable progress in the detection, protection and decontamination of biological warfare agents as many instrumentation platforms and detection methodologies are developed and commissioned. even then the threat of biological warfare agents and their use in bioterrorist attacks still remain a leading cause of global concern ...	2010	21829313
data integration for dynamic and sustainable systems biology resources: challenges and lessons learned.	systems-biology and infectious-disease (host-pathogen-environment) research and development is becoming increasingly dependent on integrating data from diverse and dynamic sources. maintaining integrated resources over long periods of time presents distinct challenges. this review describes experiences and lessons learned from integrating data in two five-year projects focused on pathosystems biology: the pathosystems resource integration center (patric, http://patric.vbi.vt.edu/), with a goal o ...	2010	20491070
pantothenate kinase from the thermoacidophilic archaeon picrophilus torridus.	pantothenate kinase (coaa) catalyzes the first step of the coenzyme a (coa) biosynthetic pathway and controls the intracellular concentrations of coa through feedback inhibition in bacteria. an alternative enzyme found in archaea, pantoate kinase, is missing in the order thermoplasmatales. the pto0232 gene from picrophilus torridus, a thermoacidophilic euryarchaeon, is shown to be a distant homologue of the prokaryotic type i coaa. the cloned gene clearly complements the poor growth of the tempe ...	2010	19854913
pantothenate kinase from the thermoacidophilic archaeon picrophilus torridus.	pantothenate kinase (coaa) catalyzes the first step of the coenzyme a (coa) biosynthetic pathway and controls the intracellular concentrations of coa through feedback inhibition in bacteria. an alternative enzyme found in archaea, pantoate kinase, is missing in the order thermoplasmatales. the pto0232 gene from picrophilus torridus, a thermoacidophilic euryarchaeon, is shown to be a distant homologue of the prokaryotic type i coaa. the cloned gene clearly complements the poor growth of the tempe ...	2010	19854913
evaluation of particulate acellular vaccines against brucella ovis infection in rams.	the attenuated brucella melitensis rev 1 vaccine, used against brucellosis infection, interferes with serological diagnosis tests, may induce abortions in pregnant animals, and may infect humans. in order to overcome these drawbacks, we developed acellular vaccines based on a brucella ovis antigenic complex (hs) containing outer membrane proteins and r-lps entrapped in poly(anhydride) conventional and mannosylated nanoparticles (np-hs and man-np-hs) or in poly(epsilon-caprolactone) microparticle ...	2010	19887131
brucella canis infection in dogs attended in veterinary clinics from patos, parabaība state, brazil.	to determine the frequency of anti-brucella canis antibodies in dogs attended in veterinary clinics from patos, paraíba state, brazil, as well as to identify risk factors and to isolate and identify the agent, 193 dogs were used. agar gel immunodiffusion test (agid) was used to detect b. canis antibodies in sera. isolation of b. canis was carried out in blood and bone marrow from seropositive animals. six animals tested seropositive in agid, resulting in a frequency of 3.11%. b. canis was isolat ...	2011	24031770
course of infection in immunocompromised mice with the emergent pathogen brucella microti.	a new brucella species brucella microti has been isolated from wild rodents and found to be pathogenic in mice. the biological relevance of this new mouse pathogen is clear as it allows us to study brucella infection in a species-specific model. the course of infection in wild type (wt) and immunodeficient mice that lack b (jh), t and b (scid) or t, b and nk (scid.beige) cells was analysed during three weeks. wt mice completely cleared bacteria from liver and spleen after that time. however, sci ...	2011	21825066
annotation of protein domains reveals remarkable conservation in the functional make up of proteomes across superkingdoms.	the functional repertoire of a cell is largely embodied in its proteome, the collection of proteins encoded in the genome of an organism. the molecular functions of proteins are the direct consequence of their structure and structure can be inferred from sequence using hidden markov models of structural recognition. here we analyze the functional annotation of protein domain structures in almost a thousand sequenced genomes, exploring the functional and structural diversity of proteomes. we find ...	2011	24710297
leptospirosis as the most frequent infectious disease impairing productivity in small ruminants in rio de janeiro, brazil.	despite the importance of small ruminants breeding in developing countries, milk/meat productivity remains unsatisfactory. infectious diseases, such as leptospirosis, brucellosis, and small ruminant lentiviruses (srlvs), contribute to this scenario. the objective of the present study was to determine the role of each of these diseases in the productivity of small ruminants breeding in rio de janeiro, brazil. in goats, 343 samples were tested for leptospirosis, 560 for brucella abortus, and 506 f ...	2011	21898182
brucellosis ontology (idobru) as an extension of the infectious disease ontology.	abstract:	2011	22041276
revised genome sequence of brucella suis 1330.	brucella suis is a causative agent of porcine brucellosis. we report the resequencing of the original sample upon which the published sequence of brucella suis 1330 is based and describe the differences between the published assembly and our assembly at 12 loci.	2011	22038969
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 ...	2011	22155231
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 ...	2011	22155231
PATRIC: the comprehensive bacterial bioinformatics resource with a focus on human pathogenic species.	Funded by the National Institute of Allergy and Infectious Diseases, the Pathosystems Resource Integration Center (PATRIC) is a genomics-centric relational database and bioinformatics resource designed to assist scientists in infectious-disease research. Specifically, PATRIC provides scientists with (i) a comprehensive bacterial genomics database, (ii) a plethora of associated data relevant to genomic analysis, and (iii) an extensive suite of computational tools and platforms for bioinformatics ...	2011	21896772
Reliable identification at the species level of Brucella isolates with MALDI-TOF-MS.	ABSTRACT: BACKGROUND: The genus Brucella contains highly infectious species that are classified as biological threat agents. The timely detection and identification of the microorganism involved is essential for an effective response not only to biological warfare attacks but also to natural outbreaks. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is a rapid method for the analysis of biological samples. The advantages of this method, compared to co ...	2011	22192890
Intra-species biodiversity of the genetically homologous species Brucella microti.	Brucellosis is one of the major bacterial zoonoses worldwide. In the last decade, an increasing number of atypical Brucella strains and species have been described. Brucella microti in particular attracted attention because this species not only infects mammalian hosts but also persists in soil. An environmental reservoir may pose a new public health risk leading to the re-emergence of brucellosis. In a polyphasic approach, comprising conventional microbiological, extensive biochemical and molec ...	2011	22210211
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 ...	2011	22216095
ribosomal rna sequence analysis of brucella infection misidentified as ochrobactrum anthropi infection.	a brucella isolate was identified from purulent material collected during a hip surgery. two previous blood cultures from the same patient yielded ochrobactrum anthropi. after rrna sequencing, all the isolates were identified as brucella species and subsequently serotyped as brucella suis. misidentification of brucella species remains a problem with bacterial identification systems.	2011	21209167
control of brucella ovis infection in sheep.	approach to control of brucella ovis would vary in different countries and areas depending on farm and flock characteristics and economic factors. eradication by a test-and-slaughter approach is the most desirable option in areas where it is logistically and financially feasible. vaccination is used in areas with a high incidence of infection where eradication would be difficult. voluntary accreditation programs have been established in some countries and are of particular benefit to pedigree ra ...	2011	21215890
development of a selective culture medium for primary isolation of the main brucella species.	bacteriological diagnosis of brucellosis is performed by culturing animal samples directly on both farrell medium (fm) and modified thayer-martin medium (mtm). however, despite inhibiting most contaminating microorganisms, fm also inhibits the growth of brucella ovis and some b. melitensis and b. abortus strains. in contrast, mtm is adequate for growth of all brucella species but only partially inhibitory for contaminants. moreover, the performance of both culture media for isolating b. suis has ...	2011	21270216
putative atp-binding cassette transporter is essential for brucella ovis pathogenesis in mice.	brucella ovis is a major cause of reproductive failure in sheep, which is associated with epididymitis and infertility in rams. importantly, b. ovis is one of the few brucella species that is not zoonotic. due to the scarcity of studies on b. ovis infection, a murine model of infection was developed. the roles of b. ovis genes encoding a putative hemagglutinin and an abc transporter were investigated in the mouse model. the kinetics of b. ovis infection were similar in balb/c and c57bl/6 mice, a ...	2011	21300772
laboratory animal models for brucellosis research.	brucellosis is a chronic infectious disease caused by brucella spp., a gram-negative facultative intracellular pathogen that affects humans and animals, leading to significant impact on public health and animal industry. human brucellosis is considered the most prevalent bacterial zoonosis in the world and is characterized by fever, weight loss, depression, hepato/splenomegaly, osteoarticular, and genital infections. relevant aspects of brucella pathogenesis have been intensively investigated in ...	2011	21403904
a species independent universal bio-detection microarray for pathogen forensics and phylogenetic classification of unknown microorganisms.	the ability to differentiate a bioterrorist attack or an accidental release of a research pathogen from a naturally occurring pandemic or disease event is crucial to the safety and security of this nation by enabling an appropriate and rapid response. it is critical in samples from an infected patient, the environment, or a laboratory to quickly and accurately identify the precise pathogen including natural or engineered variants and to classify new pathogens in relation to those that are known. ...	2011	21672191
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 ...	2011	21768283
identification of the virb operon genes encoding the type iv secretion system, in colombian brucella canis isolates.	canine brucellosis is a zoonotic disease caused by brucella canis. the establishment of intracellular replicative niches of b. canis is mediated by proteins secreted by the type iv secretion system, which is encoded by the virb operon. the characterization of such genes has been conducted in other species of the genus, but not in b. canis. we report the design of a multiplex pcr test for the detection of the virb operon genes of b. canis. primers for each of the 12 genes were designed and evalua ...	2012	23290573
cell mediated immune response after challenge in omp25 liposome immunized mice contributes to protection against virulent brucella abortus 544.	brucellosis is a disease affecting various domestic and wild life species, and is caused by a bacterium brucella. keeping in view the serious economic and medical consequences of brucellosis, efforts have been made to prevent the infection through the use of vaccines. cell-mediated immune responses [cmi] involving interferon gamma and cytotoxic cd4(+) and cd8(+) t cells are required for removal of intracellular brucella. omp25 has been reported to be involved in virulence of brucella melitensis, ...	2012	23273966
yersinia enterocolitica serotype o:9 cultured from swedish sheep showing serologically false-positive reactions for brucella melitensis.	in a herd of 20 sheep in sweden, a country where brucellosis has never been diagnosed in sheep or goats, a total of six sheep were found serologically positive to brucella melitensis in two different rounds of sampling. yersinia enterocolitica serotype o:9 could at the time of the second sampling be isolated from four sheep, one of them at the same time serologically positive for b. melitensis. the article describes the case and gives some background information on brucellosis and y. enterocolit ...	2012	23240071
the histidine kinase pdhs controls cell cycle progression of the pathogenic alphaproteobacterium brucella abortus.	bacterial differentiation is often associated with the asymmetric localization of regulatory proteins, such as histidine kinases. pdhs is an essential and polarly localized histidine kinase in the pathogenic alphaproteobacterium brucella abortus. after cell division, pdhs is asymmetrically segregated between the two sibling cells, highlighting a differentiation event. however, the function(s) of pdhs in the b. abortus cell cycle remains unknown. we used an original approach, the pentapeptide sca ...	2012	22843843
comparative genomics of early-diverging brucella strains reveals a novel lipopolysaccharide biosynthesis pathway.	brucella species are gram-negative bacteria that infect mammals. recently, two unusual strains (brucella inopinata bo1(t) and b. inopinata-like bo2) have been isolated from human patients, and their similarity to some atypical brucellae isolated from australian native rodent species was noted. here we present a phylogenomic analysis of the draft genome sequences of bo1(t) and bo2 and of the australian rodent strains 83-13 and nf2653 that shows that they form two groups well separated from the ot ...	2012	22930339
improved variation calling via an iterative backbone remapping and local assembly method for bacterial genomes.	sequencing data analysis remains limiting and problematic, especially for low complexity repeat sequences and transposon elements due to inherent sequencing errors and short sequence read lengths. we have developed a program, reviseq, which uses a hybrid method composed of iterative remapping and local assembly upon a bacterial sequence backbone. application of this method to six brucella suis field isolates compared to the newly revised b. suis 1330 reference genome identified on average 13, 15 ...	2012	22967795
sinorhizobium meliloti exor is the target of periplasmic proteolysis.	sinorhizobium meliloti exor regulates the production of succinoglycan and flagella through the exos/chvi two-component regulatory system. exor has been proposed to inhibit the exos sensor through direct interaction in the periplasm. to understand how exor suppression of exos is relieved, which is required for the expression of exos/chvi-regulated symbiosis genes, we characterized wild-type exor and exor95 mutant proteins. in addition to the previously identified precursor and mature forms of exo ...	2012	22636773
comparison of two suspension arrays for simultaneous detection of five biothreat bacterial in powder samples.	we have developed novel bio-plex assays for simultaneous detection of bacillus anthracis, yersinia pestis, brucella spp., francisella tularensis, and burkholderia pseudomallei. universal primers were used to amplify highly conserved region located within the 16s rrna amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms. the other assay is based on multiplex pcr to simultaneously amplify five species-specific pathogen identification-targeted regi ...	2012	22690123
metal acquisition and virulence in brucella.	similar to other bacteria, brucella strains require several biologically essential metals for their survival in vitro and in vivo. acquiring sufficient levels of some of these metals, particularly iron, manganese and zinc, is especially challenging in the mammalian host, where sequestration of these micronutrients is a well-documented component of both the innate and acquired immune responses. this review describes the brucella metal transporters that have been shown to play critical roles in th ...	2012	22632611
a comparative genomics screen identifies a sinorhizobium meliloti 1021 sodm-like gene strongly expressed within host plant nodules.	we have used the genomic data in the integrated microbial genomes system of the department of energy's joint genome institute to make predictions about rhizobial open reading frames that play a role in nodulation of host plants. the genomic data was screened by searching for orfs conserved in α-proteobacterial rhizobia, but not conserved in closely-related non-nitrogen-fixing α-proteobacteria.	2012	22587634
spontaneous excision of the o-polysaccharide wbka glycosyltranferase gene is a cause of dissociation of smooth to rough brucella colonies.	the brucellae are gram-negative pathogens that cause brucellosis, a zoonosis of worldwide importance. the genus brucella includes smooth and rough species that differ in that they carry smooth and rough lipopolysaccharides, respectively. brucella abortus, b. melitensis, and b. suis are typical smooth species. however, these smooth brucellae dissociate into rough mutants devoid of the lipopolysaccharide o-polysaccharide, a major antigen and a virulence determinant encoded in regions wbo (included ...	2012	22328663
vaxjo: a web-based vaccine adjuvant database and its application for analysis of vaccine adjuvants and their uses in vaccine development.	vaccine adjuvants are compounds that enhance host immune responses to co-administered antigens in vaccines. vaxjo is a web-based central database and analysis system that curates, stores, and analyzes vaccine adjuvants and their usages in vaccine development. basic information of a vaccine adjuvant stored in vaxjo includes adjuvant name, components, structure, appearance, storage, preparation, function, safety, and vaccines that use this adjuvant. reliable references are curated and cited. bioin ...	2012	22505817
brucella ceti and brucellosis in cetaceans.	since the first case of brucellosis detected in a dolphin aborted fetus, an increasing number of brucella ceti isolates has been reported in members of the two suborders of cetaceans: mysticeti and odontoceti. serological surveys have shown that cetacean brucellosis may be distributed worldwide in the oceans. although all b. ceti isolates have been included within the same species, three different groups have been recognized according to their preferred host, bacteriological properties, and dist ...	2012	22919595
analysis of the sos response of vibrio and other bacteria with multiple chromosomes.	the sos response is a well-known regulatory network present in most bacteria and aimed at addressing dna damage. it has also been linked extensively to stress-induced mutagenesis, virulence and the emergence and dissemination of antibiotic resistance determinants. recently, the sos response has been shown to regulate the activity of integrases in the chromosomal superintegrons of the vibrionaceae, which encompasses a wide range of pathogenic species harboring multiple chromosomes. here we combin ...	2012	22305460
analyses of brucella pathogenesis, host immunity, and vaccine targets using systems biology and bioinformatics.	brucella is a gram-negative, facultative intracellular bacterium that causes zoonotic brucellosis in humans and various animals. out of 10 classified brucella species, b. melitensis, b. abortus, b. suis, and b. canis are pathogenic to humans. in the past decade, the mechanisms of brucella pathogenesis and host immunity have been extensively investigated using the cutting edge systems biology and bioinformatics approaches. this article provides a comprehensive review of the applications of omics ...	2012	22919594
conjunctival vaccination against brucella ovis in mice with mannosylated nanoparticles.	the use of sub-unit vaccines can solve some drawbacks associated with traditional attenuated or inactivated ones. however, in order to improve their immunogenicity, these vaccines needs to be associated to an appropriate adjuvant which, adequately selected, may also offer an alternative pathway for administration. the aim of this work was to evaluate the protection offered by the hot saline complex extracted from brucella ovis (hs) encapsulated in mannosylated nanoparticles (man-np-hs) when inst ...	2012	22846987
[new pharmaceutical dosage forms for allergy treatment].	specific immunotherapy involves certain drawbacks which could be minimized by the use of appropriate adjuvants, capable of amplifying the right immune response with minimal side effects. in this context, we review different types of immunotherapy vehicles and coadjuvants. we describe previous studies by our group in which we demonstrated the adjuvant capacity of gantrez® an nanoparticles, which can effectively enhance the immune response. we employed two types of nanoparticles (with and without ...	2012	22552127
the virb-encoded type iv secretion system is critical for establishment of infection and persistence of brucella ovis infection in mice.	brucella spp. are gram-negative intracellular bacterial pathogens that cause chronic infections. brucella virulence factors include a type iv secretion system (t4ss) and its lipopolysaccharide (lps), which are essential for persistence. however, the role of the virb-encoded t4ss has not been investigated in naturally rough brucella species such as brucella ovis. in this study, male 6-week old balbc mice were infected with b. ovis, brucella abortus, and their respective δvirb2 mutant strains. dur ...	2012	22483850
factors associated with failure in breeding soundness examination of western usa rams.	breeding-soundness examination (bse) and eradication of brucella ovis infection in rams are critical components of flock-health programs. the aims of this retrospective, cross-sectional study were to describe the results of bse in a large sample of rams in the western usa and to determine the association between bse outcome and the semen collection method (penis manually extended vs. retained in the preputial cavity), ram body-condition score (bcs), the presence of ulcerative posthitis, and the ...	2012	22385780
pathology of brucella ovis infection in red deer stags (cervus elaphus).	to describe the pathology of the reproductive tract of red deer stags with active brucella ovis infection and in stags in which b. ovis infection had resolved.	2012	22352933
quorum-sensing and bvrr/bvrs regulation, the type iv secretion system, cyclic glucans, and baca in the virulence of brucella ovis: similarities to and differences from smooth brucellae.	brucella ovis is a rough bacterium--lacking o-polysaccharide chains in the lipopolysaccharide--that is virulent in its natural host and whose virulence mechanisms remain almost unexplored. in a search for additional traits that distinguish b. ovis from smooth brucella, which require o-polysaccharide chains for virulence, we have analyzed the significance in b. ovis of the main virulence factors described for smooth brucella. attempts to obtain strains of virulent b. ovis strain pa that are mutat ...	2012	22392933
from environment to man: genome evolution and adaptation of human opportunistic bacterial pathogens.	environment is recognized as a huge reservoir for bacterial species and a source of human pathogens. some environmental bacteria have an extraordinary range of activities that include promotion of plant growth or disease, breakdown of pollutants, production of original biomolecules, but also multidrug resistance and human pathogenicity. the versatility of bacterial life-style involves adaptation to various niches. adaptation to both open environment and human specific niches is a major challenge ...	2012	24704914
development of a novel genus-specific real-time pcr assay for detection and differentiation of bartonella species and genotypes.	the genus bartonella includes numerous species with varied host associations, including several that infect humans. development of a molecular diagnostic method capable of detecting the diverse repertoire of bartonella species while maintaining genus specificity has been a challenge. we developed a novel real-time pcr assay targeting a 301-bp region of the ssra gene of bartonella and demonstrated specific amplification in over 30 bartonella species, subspecies, and strains. subsequent analysis o ...	2012	22378904
effects of resource chemistry on the composition and function of stream hyporheic biofilms.	fluvial ecosystems process large quantities of dissolved organic matter as it moves from the headwater streams to the sea. in particular, hyporheic sediments are centers of high biogeochemical reactivity due to their elevated residence time and high microbial biomass and activity. however, the interaction between organic matter and microbial dynamics in the hyporheic zone remains poorly understood. we evaluated how variance in resource chemistry affected the microbial community and its associate ...	2012	22347877
assessment of the diagnostic sensitivity and specificity of an indirect elisa kit for the diagnosis of brucella ovis infection in rams.	brucella ovis causes an infectious disease responsible for infertility and subsequent economic losses in sheep production. the standard serological test to detect b. ovis infection in rams is the complement fixation test (cft), which has imperfect sensitivity and specificity in addition to technical drawbacks. other available tests include the indirect enzyme-linked immunosorbent assays (i-elisa) but no i-elisa kit has been fully evaluated.the study aimed to compare an i-elisa kit and the standa ...	2012	22640401
characterization of species-specific repeats in 613 prokaryotic species.	prokaryotes are in general believed to possess small, compactly organized genomes, with repetitive sequences forming only a small part of them. nonetheless, many prokaryotic genomes in fact contain species-specific repeats (>85 bp long genomic sequences with less than 60% identity to other species) as we have previously demonstrated. however, it is not known at present how frequent such species-specific repeats are and what their functional roles in bacterial genomes may be. therefore, we have c ...	2012	22368180
evolutionary diversification and characterization of the eubacterial gene family encoding dxr type ii, an alternative isoprenoid biosynthetic enzyme.	isoprenoids constitute a vast family of natural compounds performing diverse and essential functions in all domains of life. in most eubacteria, isoprenoids are synthesized through the methylerythritol 4-phosphate (mep) pathway. the production of mep is usually catalyzed by deoxyxylulose 5-phosphate reductoisomerase (dxr-i) but a few organisms use an alternative dxr-like enzyme (dxr-ii).	2013	24004839
brucella bior regulator defines a complex regulatory mechanism for bacterial biotin metabolism.	the enzyme cofactor biotin (vitamin h or b7) is an energetically expensive molecule whose de novo biosynthesis requires 20 atp equivalents. it seems quite likely that diverse mechanisms have evolved to tightly regulate its biosynthesis. unlike the model regulator bira, a bifunctional biotin protein ligase with the capability of repressing the biotin biosynthetic pathway, bior has been recently reported by us as an alternative machinery and a new type of gntr family transcriptional factor that ca ...	2013	23729648
bacterial endosymbiosis in a chordate host: long-term co-evolution and conservation of secondary metabolism.	intracellular symbiosis is known to be widespread in insects, but there are few described examples in other types of host. these symbionts carry out useful activities such as synthesizing nutrients and conferring resistance against adverse events such as parasitism. such symbionts persist through host speciation events, being passed down through vertical transmission. due to various evolutionary forces, symbionts go through a process of genome reduction, eventually resulting in tiny genomes wher ...	2013	24324632
functional genes to assess nitrogen cycling and aromatic hydrocarbon degradation: primers and processing matter.	targeting sequencing to genes involved in key environmental processes, i.e., ecofunctional genes, provides an opportunity to sample nature's gene guilds to greater depth and help link community structure to process-level outcomes. vastly different approaches have been implemented for sequence processing and, ultimately, for taxonomic placement of these gene reads. the overall quality of next generation sequence analysis of functional genes is dependent on multiple steps and assumptions of unknow ...	2013	24062736
species-specific multiplex pcr for the diagnosis of brucella ovis, actinobacillus seminis, and histophilus somni infection in rams.	infectious ovine epididymitis results in substantial economic losses worldwide due to reproductive failure and culling of breeders. the most common causative agents of these infections are brucella ovis, actinobacillus seminis, and histophilus somni. the aim of this study was to develop a multiplex pcr assay for simultaneous detection of brucella ovis, actinobacillus seminis, and histophilus somni with species-specific primers applied to biological samples for molecular diagnosis of these infect ...	2013	23514236
applications of polymeric adjuvants in studying autoimmune responses and vaccination against infectious diseases.	polymers as an adjuvant are capable of enhancing the vaccine potential against various infectious diseases and also are being used to study the actual autoimmune responses using self-antigen(s) without involving any major immune deviation. several natural polysaccharides and their derivatives originating from microbes and plants have been tested for their adjuvant potential. similarly, numerous synthetic polymers including polyelectrolytes, polyesters, polyanhydrides, non-ionic block copolymers ...	2013	23173193
rough virulent strain of brucella ovis induces pro- and anti-inflammatory cytokines in reproductive tissues in experimentally infected rams.	the ovine brucellosis caused by brucella ovis has tropism for reproductive tissues but until now the mechanism of bacterial persistence is not understood. cytokine expression profiles were studied for 8 months in rams after being experimentally infected with the rough virulent strain of b. ovis (r-b. ovis) to study the pathogenesis of b. ovis and immune mechanism possibly associated to bacteria tropism and persistence. the messenger rna (mrna) expression levels of interleukin-1α (il-1α), il-1β, ...	2013	23041076
a duplex pcr for rapid and simultaneous detection of brucella spp. in human blood samples.	to design a duplex pcr for rapid and simultaneous detection of brucella species. in human blood samples.	2013	23711705
natural is711 insertion causing omp31 gene inactivation in brucella ovis.	the present report describes an atypical brucella ovis strain (bo10) isolated from the epididymis and testis of an infected ram. macroscopic and microscopic lesions characteristic for the infection, including positive brucella immunostaining, were observed within lesions in the genital organs. compared to other isolates, strain bo10 required an additional day (a total of 96 hr) of incubation to form visible colonies, showed a distinct carbon source utilization profile, agglutinated only weakly w ...	2013	23404477
immune response and serum bactericidal activity against brucella ovis elicited using a short immunization schedule with the polymeric antigen blsomp31 in rams.	brucella ovis is the etiologic agent of ovine brucellosis. the control measures for this disease are periodical diagnosis by serological tests and/or bacteriological culture and culling of positive animals. vaccination with brucella melitensis rev 1 is recommended when prevalence is high. this attenuated strain vaccine gives protection against b. ovis but it has important disadvantages associated with the development of antibodies interfering with serodiagnosis, virulence for humans and the proh ...	2013	23643287
clinical, culture, serology, and histopathology outcomes of bighorn sheep experimentally infected with brucella ovis.	disease caused by brucella ovis has not been previously reported in bighorn sheep (bhs; ovis canadensis canadensis). antibodies to b. ovis, however, are occasionally detected in free-ranging bhs, and this has been a concern for managers involved in translocation programs. to investigate the pathogenesis of b. ovis infection in this species, 20 bhs (10 male, 10 female) were inoculated intraconjunctivally (ic) with 5.4 × 10(8) colony forming units (cfu) b. ovis. six bhs (three male, three female) ...	2013	24502717
a lysozyme-like protein in brucella abortus is involved in the early stages of intracellular replication.	secretion of proteins in gram-negative bacteria is a high-energy-consuming process that requires translocation across two membranes and a periplasmic space composed of a mesh-like layer, the peptidoglycan. to achieve this, bacteria have evolved complex secretion systems that cross these barriers, and in many cases there are specific peptidoglycanases that degrade the peptidoglycan to allow the proper assembly of the secretion machinery. we describe here the identification and characterization of ...	2013	23319555
molecular mechanisms for the subversion of myd88 signaling by tcpc from virulent uropathogenic escherichia coli.	the toll/il-1 receptor (tir) domains are crucial signaling modules during innate immune responses involving the toll-like receptors (tlrs) and il-1 receptor (il-1r). myeloid differential factor 88 (myd88) is a central tir domain-containing adapter molecule responsible for nearly all tlr-mediated signaling and is targeted by a tir domain-containing protein c (tcpc) from virulent uropathogenic escherichia coli, a common human pathogen. the mechanism of such molecular antagonism has remained elusiv ...	2013	23569230
crystallization of the acyl-coa thioesterase tesb from yersinia pestis.	yersinia pestis is a highly virulent human pathogen and is the causative agent of bubonic plague. spread through the bite of infected fleas, plague epidemics have marked important events in history, including the justinian plague (6th century), the black death (14th century) which decimated nearly one quarter of the european population, and more recently the orientalis plague (1894). to date, deaths are still being reported and, without treatment, the disease kills most people within 4 days. one ...	2013	23385765
in vivo differences in the virulence, pathogenicity, and induced protective immunity of wboa mutants from genetically different parent brucella spp.	to explore the effects of the genetic background on the characteristics of wboa gene deletion rough mutants generated from different parent brucella sp. strains, we constructed the rough-mutant strains brucella melitensis 16 m-mb6, b. abortus 2308-sb6, b. abortus s19-rb6, and b. melitensis ni-nb6 and evaluated their survival, pathogenicity, and induced protective immunity in mice and sheep. in mice, the survival times of the four mutants were very different in the virulence assay, from less than ...	2013	23239800
brucella ovis lacking a species-specific putative atp-binding cassette transporter is attenuated but immunogenic in rams.	ovine brucellosis caused by brucella ovis is considered one of the most important reproductive diseases of rams worldwide. this study aimed to characterize the kinetics of infection of a δabcab b. ovis mutant strain in rams. twelve 1-year-old crossbred rams were used. six rams were challenged with 2 ml of a suspension containing 1.2×10(9) cfu/ml of b. ovis strain atcc25840 (wild type) by intraprepucial inoculation and additional 50 μl in each conjunctival sac of a suspension containing 1.2×10(10 ...	2013	24075357
[immune response and reproductive consequences in experimentally infected ewes with brucella ovis during late pregnancy].	ovine brucellosis by brucella ovis is a highly prevalent disease in argentina. this study aimed to evaluate the pathogenicity of b. ovis and the serological response in ewes during late pregnancy and in their offspring. six adult ewes were distributed in two groupsg1 (pregnant females, n = 4) and g2 (nonpregnant females, n = 2). three pregnant ewes at 15 days prepartum and one nonpregnant eve were inoculated with b. ovis. sera of sheep and their offspring were analyzed by different serological t ...	2013	23560783
[radiolabeling and biodistribution studies of polymeric nanoparticles as adjuvants for ocular vaccination against brucellosis].	to optimize radiolabeling with (99m)tc of mannosylated gantrez(®) nanoparticles loaded with the brucella ovis antigen (man-np-hs) and to carry out biodistribution studies in mice after ocular administration of the nanoparticles.	2013	23332663