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Sample records for abortus ure2 region

  1. Brucella abortus ure2 region contains an acid-activated urea transporter and a nickel transport system

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    García-Lobo Juan M

    2010-04-01

    Full Text Available Abstract Background Urease is a virulence factor that plays a role in the resistance of Brucella to low pH conditions, both in vivo and in vitro. Brucella contains two separate urease gene clusters, ure1 and ure2. Although only ure1 codes for an active urease, ure2 is also transcribed, but its contribution to Brucella biology is unknown. Results Re-examination of the ure2 locus showed that the operon includes five genes downstream of ureABCEFGDT that are orthologs to a nikKMLQO cluster encoding an ECF-type transport system for nickel. ureT and nikO mutants were constructed and analyzed for urease activity and acid resistance. A non-polar ureT mutant was unaffected in urease activity at neutral pH but showed a significantly decreased activity at acidic pH. It also showed a decreased survival rate to pH 2 at low concentration of urea when compared to the wild type. The nikO mutant had decreased urease activity and acid resistance at all urea concentrations tested, and this phenotype could be reverted by the addition of nickel to the growth medium. Conclusions Based on these results, we concluded that the operon ure2 codes for an acid-activated urea transporter and a nickel transporter necessary for the maximal activity of the urease whose structural subunits are encoded exclusively by the genes in the ure1 operon.

  2. Evolutionary divergence of Ure2pA glutathione transferases in wood degrading fungi.

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    Roret, Thomas; Thuillier, Anne; Favier, Frédérique; Gelhaye, Eric; Didierjean, Claude; Morel-Rouhier, Mélanie

    2015-10-01

    The intracellular systems of detoxification are crucial for the survival of wood degrading fungi. Within these systems, glutathione transferases could play a major role since this family of enzymes is specifically extended in lignolytic fungi. In particular the Ure2p class represents one third of the total GST number in Phanerochaete chrysosporium. These proteins have been phylogenetically split into two subclasses called Ure2pA and Ure2pB. Ure2pB can be classified as Nu GSTs because of shared structural and functional features with previously characterized bacterial isoforms. Ure2pA can rather be qualified as Nu-like GSTs since they exhibit a number of differences. Ure2pA possess a classical transferase activity, a more divergent catalytic site and a higher structural flexibility for some of them, compared to Nu GSTs. The characterization of four members of this Ure2pA subclass (PcUre2pA4, PcUre2pA5, PcUre2pA6 and PcUre2pA8) revealed specific functional and structural features, suggesting that these enzymes have rapidly evolved and differentiated, probably to adapt to the complex chemical environment associated with wood decomposition.

  3. Functional Diversification of Fungal Glutathione Transferases from the Ure2p Class

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    Anne Thuillier

    2011-01-01

    Full Text Available The glutathione-S-transferase (GST proteins represent an extended family involved in detoxification processes. They are divided into various classes with high diversity in various organisms. The Ure2p class is especially expanded in saprophytic fungi compared to other fungi. This class is subdivided into two subclasses named Ure2pA and Ure2pB, which have rapidly diversified among fungal phyla. We have focused our analysis on Basidiomycetes and used Phanerochaete chrysosporium as a model to correlate the sequence diversity with the functional diversity of these glutathione transferases. The results show that among the nine isoforms found in P. chrysosporium, two belonging to Ure2pA subclass are exclusively expressed at the transcriptional level in presence of polycyclic aromatic compounds. Moreover, we have highlighted differential catalytic activities and substrate specificities between Ure2pA and Ure2pB isoforms. This diversity of sequence and function suggests that fungal Ure2p sequences have evolved rapidly in response to environmental constraints.

  4. Functional diversification of fungal glutathione transferases from the ure2p class.

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    Thuillier, Anne; Ngadin, Andrew A; Thion, Cécile; Billard, Patrick; Jacquot, Jean-Pierre; Gelhaye, Eric; Morel, Mélanie

    2011-01-01

    The glutathione-S-transferase (GST) proteins represent an extended family involved in detoxification processes. They are divided into various classes with high diversity in various organisms. The Ure2p class is especially expanded in saprophytic fungi compared to other fungi. This class is subdivided into two subclasses named Ure2pA and Ure2pB, which have rapidly diversified among fungal phyla. We have focused our analysis on Basidiomycetes and used Phanerochaete chrysosporium as a model to correlate the sequence diversity with the functional diversity of these glutathione transferases. The results show that among the nine isoforms found in P. chrysosporium, two belonging to Ure2pA subclass are exclusively expressed at the transcriptional level in presence of polycyclic aromatic compounds. Moreover, we have highlighted differential catalytic activities and substrate specificities between Ure2pA and Ure2pB isoforms. This diversity of sequence and function suggests that fungal Ure2p sequences have evolved rapidly in response to environmental constraints.

  5. Functional Diversification of Fungal Glutathione Transferases from the Ure2p Class

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    Anne Thuillier; Ngadin, Andrew A.; Cécile Thion; Patrick Billard; Jean-Pierre Jacquot; Eric Gelhaye; Mélanie Morel

    2011-01-01

    The glutathione-S-transferase (GST) proteins represent an extended family involved in detoxification processes. They are divided into various classes with high diversity in various organisms. The Ure2p class is especially expanded in saprophytic fungi compared to other fungi. This class is subdivided into two subclasses named Ure2pA and Ure2pB, which have rapidly diversified among fungal phyla. We have focused our analysis on Basidiomycetes and used Phanerochaete chrysosporium as a model to c...

  6. Seroprevalence of Chlamydia abortus in camel in the western region of Libya

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    Rabia Elzlitne

    2016-06-01

    Conclusion: The present findings signify the C. abortus as a potential agent to cause abortion in Libyan camel (C. dromedarius. Besides, the persons who handle camels in Libya are at risk of infecting with C. abortus. [J Adv Vet Anim Res 2016; 3(2.000: 178-183

  7. Genome Sequences of 11 Brucella abortus Isolates from Persistently Infected Italian Regions

    OpenAIRE

    2015-01-01

    Bovine brucellosis, typically caused by Brucella abortus, has been eradicated from much of the developed world. However, the disease remains prevalent in southern Italy, persisting as a public and livestock health concern. We report here the whole-genome sequences of 11 isolates from cattle (Bos taurus) and water buffalo (Bubalus bubalis) that are representative of the current genetic diversity of B. abortus lineages circulating in Italy.

  8. Atypical features of a Ure2p glutathione transferase from Phanerochaete chrysosporium.

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    Thuillier, Anne; Roret, Thomas; Favier, Frédérique; Gelhaye, Eric; Jacquot, Jean-Pierre; Didierjean, Claude; Morel-Rouhier, Mélanie

    2013-07-11

    Glutathione transferases (GSTs) are known to transfer glutathione onto small hydrophobic molecules in detoxification reactions. The GST Ure2pB1 from Phanerochaete chrysosporium exhibits atypical features, i.e. the presence of two glutathione binding sites and a high affinity towards oxidized glutathione. Moreover, PcUre2pB1 is able to efficiently deglutathionylate GS-phenacylacetophenone. Catalysis is not mediated by the cysteines of the protein but rather by the one of glutathione and an asparagine residue plays a key role in glutathione stabilization. Interestingly PcUre2pB1 interacts in vitro with a GST of the omega class. These properties are discussed in the physiological context of wood degrading fungi.

  9. Variables Associated with Infections of Cattle by Brucella abortus., Leptospira spp. and Neospora spp. in Amazon Region in Brazil.

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    Chiebao, D P; Valadas, S Y O B; Minervino, A H H; Castro, V; Romaldini, A H C N; Calhau, A S; De Souza, R A B; Gennari, S M; Keid, L B; Soares, R M

    2015-10-01

    The frequency of Neospora spp., Leptospira spp. and Brucella abortus infections in adult cattle was determined in herds of the State of Pará, Brazil, which is an important region for cattle production located in the Amazon region. A total of 3466 adult female cattle from 176 herds were tested, leading to a frequency of seropositive animals of 14.7%, 3.7% and 65.5% and a herd positivity of 87.4%, 41.3% and 98.8% for infections caused by Neospora spp., B. abortus and Leptospira spp., respectively. The five most frequently diagnosed serologic responses to Leptospira spp. were those against serovars hardjo, wolfii, grippotyphosa, hebdomadis and shermani. The following associations were found: practice of artificial insemination, large farm size, large herd size, large number of dogs and high number of total abortions per year with the presence of antibodies against serovar hardjo; positive results to serovar grippotyphosa with the presence of dogs; inappropriate disposal of aborted foetuses with positivity to serovar hebdomadis. Serovar grippotyphosa was also associated with number of episodes of abortions. Neospora spp. positive herds were associated with episodes of abortion and B. abortus infection with the disposal of dead animals and aborted foetuses on pastures and with the use of artificial insemination. In conclusion, the high frequency of brucellosis, leptospirosis and neosporosis in the region may be a consequence of social, natural and raising conditions as: (i) climate conditions that favour the survival and spread of pathogens in the environment; (ii) farms located in regions bordering forest areas; (iii) farms in areas of difficult access to the veterinary service; (iv) extensive beef herds raised at pastures with different age and productive groups inter-mingled; and (v) minimal concerns regarding hygiene practices and disease prevention measures.

  10. Extensive de novo solid-state NMR assignments of the 33 kDa C-terminal domain of the Ure2 prion

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    Habenstein, Birgit [UMR 5086 CNRS/Universite de Lyon 1, Institut de Biologie et Chimie des Proteines (France); Wasmer, Christian [Harvard Medical School (United States); Bousset, Luc; Sourigues, Yannick [UPR 3082 CNRS, Laboratoire d' Enzymologie et Biochimie Structurales (France); Schuetz, Anne [ETH Zurich, Physical Chemistry (Switzerland); Loquet, Antoine [Max Planck Institute for Biophysical Chemistry (Germany); Meier, Beat H., E-mail: beme@ethz.ch [ETH Zurich, Physical Chemistry (Switzerland); Melki, Ronald, E-mail: melki@lebs.cnrs-gif.fr [UPR 3082 CNRS, Laboratoire d' Enzymologie et Biochimie Structurales (France); Boeckmann, Anja, E-mail: a.bockmann@ibcp.fr [UMR 5086 CNRS/Universite de Lyon 1, Institut de Biologie et Chimie des Proteines (France)

    2011-11-15

    We present the de novo resonance assignments for the crystalline 33 kDa C-terminal domain of the Ure2 prion using an optimized set of five 3D solid-state NMR spectra. We obtained, using a single uniformly {sup 13}C, {sup 15}N labeled protein sample, sequential chemical-shift information for 74% of the N, C{alpha}, C{beta} triples, and for 80% of further side-chain resonances for these spin systems. We describe the procedures and protocols devised, and discuss possibilities and limitations of the assignment of this largest protein assigned today by solid-state NMR, and for which no solution-state NMR shifts were available. A comparison of the NMR chemical shifts with crystallographic data reveals that regions with high crystallographic B-factors are particularly difficult to assign. While the secondary structure elements derived from the chemical shift data correspond mainly to those present in the X-ray crystal structure, we detect an additional helical element and structural variability in the protein crystal, most probably originating from the different molecules in the asymmetric unit, with the observation of doubled resonances in several parts, including entire stretches, of the protein. Our results provide the point of departure towards an atomic-resolution structural analysis of the C-terminal Ure2p domain in the context of the full-length prion fibrils.

  11. Prevalence and molecular identification of Chlamydia abortus in commercial dairy goat farms in a hot region in Mexico.

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    Campos-Hernández, Eleuterio; Vázquez-Chagoyán, Juan Carlos; Salem, Abdelfattah Z M; Saltijeral-Oaxaca, Jorge Antonio; Escalante-Ochoa, Cristina; López-Heydeck, Sandra M; de Oca-Jiménez, Roberto Montes

    2014-08-01

    The aim of this study was to determine the seroprevalence and presence of Chlamydia abortus in Saanen breed female goats from commercial dairy goat farms under intensive production in the municipality of Guanajuato, Mexico. Sera were collected to determine the prevalence of anti-C. abortus IgG antibodies using recombinant enzyme-linked immunosorbent assay (rELISA) and cell culture. Polymerase chain reaction (PCR) was used to prove the presence of the pathogen in swab samples collected from the vagina and rectum of selected animals. Additionally, foetal tissue samples from a sudden abortion were collected. C. abortus prevalence in female goats of commercial milking farms sampled in Guanajuato, Mexico, was 4.87% (n = 246). Seropositive animals were found in six out of nine (66.6%) dairy goat farms sampled, and prevalence among animals in individual farms ranged between 3.44 and 13.51%. C. abortus was detected using PCR in spleen tissue from the aborted foetus. PCR-based detection, as well as isolation from vaginal and rectal swabs, was not possible in the present study. Isolation through cell culture was also unsuccessful from aborted foetal tissue samples. In conclusion, the results from rELISA and PCR show that C. abortus is present in dairy goat farms in the state of Guanajuato, Mexico.

  12. Genotipos de aislados de campo de Brucella abortus de distintas regiones geográficas de Chile Genotypes of Brucella abortus field isolates from different geographical regions of Chile

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    M Mancilla

    2008-01-01

    Full Text Available La brucelosis bovina es una enfermedad zoonótica, endémica de alto impacto económico. La identificación genética de las cepas prevalentes de Brucella abortus, el patógeno, es clave para establecer estrategias epidemiológicas de control de la enfermedad. La secuencia de inserción IS711 ha sido utilizada como un marcador genético para diferenciar entre especies de Brucella, miembros de una misma especie y dentro de un mismo biovar. Hemos analizado los perfiles de IS711-RFLP de 46 aislados de B. abortus, recolectados durante el periodo 1997-2005, provenientes de 16 áreas geográficas diferentes de Chile. Todos los aislados fueron previamente identificados como B. abortus biovar 1, utilizando las técnicas convencionales. De estos, el 87% compartieron el mismo perfil de IS711-RFLP, mientras que el 8,7% correspondió al patrón de la cepa vacuna RB51. En este trabajo se informa el hallazgo de dos cepas indistinguibles por PCR AMOS con perfiles nuevos de IS711-RFLP, no reportados previamente.Bovine brucellosis is an endemic, zoonotic disease of high economic impact. The genetic identification of the prevalent Brucella abortus strains, the pathogen, is key to pursue further epidemiological strategies for disease control. The insertion sequence IS711 has been used as genetic marker to differentiate among Brucella species, members of the same specie and within the same biovar. We have analyzed the IS711-RFLP pattern for 46 B. abortus isolates, collected during the period of 1997-2005 from 16 different geographical areas of Chile. All isolates were previously identified by conventional techniques as B. abortus biovar 1. Of these, 87% sharedthesame IS711 DNA profile, while an 8.7 % corresponded to the pattern of RB51 vaccine strain. We report the finding of two new strains, not differentiated by AMOS PCR, which showed unreported patterns of IS711-RFLP.

  13. Crystallization and preliminary X-ray crystallographic analysis of yeast prion protein Ure2p with shortened N-terminal

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    An orthorhombic crystal form of a recombinant yeast prion protein with shortened N-terminal, 90Ure2p, has been obtained. Crystals were grown by the vapordiffusion technique against a mother liquor containing imidazole. Crystals belong to the primitive orthorhombic lattice with the cell parameters a = 54.5 ?, b = 74.7 ?, c = 131.0 ?. The crystals diffract to beyond 3.0 ? resolution at a synchrotron beamline.

  14. Brucella suis urease encoded by ure1 but not ure2 is necessary for intestinal infection of BALB/c mice

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    Sriranganathan Nammalwar

    2007-06-01

    Full Text Available Abstract Background In prokaryotes, the ureases are multi-subunit, nickel-containing enzymes that catalyze the hydrolysis of urea to carbon dioxide and ammonia. The Brucella genomes contain two urease operons designated as ure1 and ure2. We investigated the role of the two Brucella suis urease operons on the infection, intracellular persistence, growth, and resistance to low-pH killing. Results The deduced amino acid sequence of urease-α subunits of operons-1 and -2 exhibited substantial identity with the structural ureases of alpha- and beta-proteobacteria, Gram-positive and Gram-negative bacteria, fungi, and higher plants. Four ure deficient strains were generated by deleting one or more of the genes encoding urease subunits of B. suis strain 1330 by allelic exchange: strain 1330Δure1K (generated by deleting ureD and ureA in ure1 operon, strain 1330Δure2K (ureB and ureC in ure2 operon, strain 1330Δure2C (ureA, ureB, and ureC in ure2 operon, and strain 1330Δure1KΔure2C (ureD and ureA in ure1 operon and ureA, ureB, and ureC in ure2 operon. When grown in urease test broth, strains 1330, 1330Δure2K and 1330Δure2C displayed maximal urease enzyme activity within 24 hours, whereas, strains 1330Δure1K and 1330Δure1KΔure2C exhibited zero urease activity even 96 h after inoculation. Strains 1330Δure1K and 1330Δure1KΔure2C exhibited slower growth rates in tryptic soy broth relative to the wild type strain 1330. When the BALB/c mice were infected intraperitoneally with the strains, six weeks after inoculation, the splenic recovery of the ure deficient strains did not differ from the wild type. In contrast, when the mice were inoculated by gavage, one week after inoculation, strain 1330Δure1KΔure2C was cleared from livers and spleens while the wild type strain 1330 was still present. All B. suis strains were killed when they were incubated in-vitro at pH 2.0. When the strains were incubated at pH 2.0 supplemented with 10 mM urea, strain

  15. The 26S Proteasome Degrades the Soluble but Not the Fibrillar Form of the Yeast Prion Ure2p In Vitro.

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    Kai Wang

    Full Text Available Yeast prions are self-perpetuating protein aggregates that cause heritable and transmissible phenotypic traits. Among these, [PSI+] and [URE3] stand out as the most studied yeast prions, and result from the self-assembly of the translation terminator Sup35p and the nitrogen catabolism regulator Ure2p, respectively, into insoluble fibrillar aggregates. Protein quality control systems are well known to govern the formation, propagation and transmission of these prions. However, little is known about the implication of the cellular proteolytic machineries in their turnover. We previously showed that the 26S proteasome degrades both the soluble and fibrillar forms of Sup35p and affects [PSI+] propagation. Here, we show that soluble native Ure2p is degraded by the proteasome in an ubiquitin-independent manner. Proteasomal degradation of Ure2p yields amyloidogenic N-terminal peptides and a C-terminal resistant fragment. In contrast to Sup35p, fibrillar Ure2p resists proteasomal degradation. Thus, structural variability within prions may dictate their ability to be degraded by the cellular proteolytic systems.

  16. Conformational change of glutathione-S-transferase by its co-expression with prion domain of yeast Ure2p

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The Ure2 protein from Saccharomyces cerevisisae has a changeable structure similar to that ofrnammalian prion protein. Its N-terminal is the prion domain (PrD) consisting of 65 amino acids which plays a critical role in yeast prion development. In this study, PrD gene was recombinated with glutathione-S-transferase(GST) gene, and a soluble GST-PrD(sGST-PrD) fusion protein was expressed in E. coli. sGST-PrD could spontaneously polymerize into amyloid fibrils in vitro, displaying typical β-sheet-type structure; it had increased resistance to proteinase K and exhibited amvloid-like optical properties. Moreover, the aggregated GST-PrD(aGST-PrD) could induce sGST-PrD to aggregate into fibrils. These results indicate that PrD could change the conformation of GST moiety in a recombinant protein with PrD to form a prion-like chimeric protein, which proves that PrD has the ability to mediate a prion-like conversion of other proteins fused with it.

  17. Efficacy of Brucella abortus vaccine strain RB51 compared to the reference vaccine Brucella abortus strain 19 in water buffalo

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    Vincenzo Caporale

    2010-03-01

    Full Text Available Approximately 250 000 water buffalo (Bubalus bubalis live in the Campania region of southern Italy where the breeding of this species is very popular. Of these animals, almost 150 000 are concentrated in the Caserta province where the prevalence of Brucella abortus in this species represents approximately 20% at herd level. The Italian brucellosis eradication programme provides a slaughter and vaccination strategy for this province. B. abortus strain RB51 (RB51 has become the official vaccine for the prevention of brucellosis in cattle in several countries. The aim of this study was to evaluate the efficacy of RB51 in water buffalo compared to the B. abortus S19 vaccine (S19. The study was performed in accordance with a protocol described in mice. Female buffalo aged five months were inoculated. Five received a RB51 dosage on two occasions that was three times greater than that approved for use in cattle and a booster after one month, five received B. abortus S19 vaccine at the standard dosage and three controls received a phosphate buffer solution. Buffalo were then challenged with a virulent B. abortus strain 544 thirty days post vaccination. Antibodies that developed in the five animals vaccinated with RB51 were not detected by the Rose Bengal test or complement fixation test (CFT and were also tested by CFT prepared with RB51 antigen. After culling, B. abortus was cultured from the spleen, retropharyngeal and supra-mammary lymph nodes. A statistical evaluation was performed to assess the immunogenicity values obtained in buffalo vaccinated with S19, compared to those obtained in buffalo vaccinated with the RB51 vaccine and in the unvaccinated control group.

  18. Biotyping and genotyping (MLVA16 of Brucella abortus isolated from cattle in Brazil, 1977 to 2008.

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    Sílvia Minharro

    Full Text Available Brucellosis is a worldwide distributed zoonosis that causes important economic losses to animal production. In Brazil, information on the distribution of biovars and genotypes of Brucella spp. is scarce or unavailable. This study aimed (i to biotype and genotype 137 Brazilian cattle isolates (from 1977 to 2008 of B. abortus and (ii to analyze their distribution. B. abortus biovars 1, 2 and 3 (subgroup 3b were confirmed and biovars 4 and 6 were first described in Brazil. Genotyping by the panel 1 revealed two groups, one clustering around genotype 40 and another around genotype 28. Panels 2A and 2B disclosed a high diversity among Brazilian B. abortus strains. Eighty-nine genotypes were found by MLVA16. MLVA16 panel 1 and 2 showed geographic clustering of some genotypes. Biotyping and MLVA16 genotyping of Brazilian B. abortus isolates were useful to better understand the epidemiology of bovine brucellosis in the region.

  19. An Engineered PrPsc-like Molecule from the Chimera of Mammalian Prion Protein and Yeast Ure2p Prion-inducing Domain

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    Shao-Man YIN; Man-Sun SY; Po TIEN

    2004-01-01

    Production of the pathogenic prion isoform prpsc-like molecules is thought to be useful forunderstanding the mysterious mechanism of conformational conversion process of prion diseases andproving the "protein-only" hypothesis. In this report, an engineered PrPsc-like conformation was producedfrom a chimera of mammalian bovine prion protein (bPrP) and yeast Ure2p prion-inducing domain (UPrD).Compared with the normal form of bPrP, the engineered recombinant protein, termed bPrP-UPrD,spontaneously aggregated into ordered fibrils under physiological condition, displaying amyloid-likecharacteristics, such as fibrillar morphology, birefringence upon binding to Congo red and increasedfluorescence intensity with Thioflavine T. Limited resistance to protease K digestion and CD spectroscopyexperiments suggested that the structure of bPrP-UPrD had been changed, and adopted a new, high contentβ-sheet conformation during the fibrils formation. Moreover, bPrP-UPrD amyloid fibrils could recruit moresoluble forms into the aggregates. Therefore, the engineered molecules could mimic significant behaviors ofPrPsc and will be helpful for further understanding the mechanism of conformational conversion process.

  20. An Engineered PrPsc-like Molecule from the Chimera of Mammalian Prion Protein and Yeast Ure2p Prion-inducing Domain

    Institute of Scientific and Technical Information of China (English)

    Shao-ManYIN; Man-SunSY; PoTIEN

    2004-01-01

    Production of the pathogenic prion isoform PrPsc-like molecules is thought to be useful forunderstanding the mysterious mechanism of conformational conversion process of prion diseases andproving the "protein-only" hypothesis. In this report, an engineered PrPsc-like conformation was producedfrom a chimera of mammalian bovine prion protein (bPrP) and yeast Ure2p prion-inducing domain (UPrD).Compared with the normal form of bPrP, the engineered recombinant protein, termed bPrP-UPrD,spontaneously aggregated into ordered fibrils under physiological condition, displaying amyloid-likecharacteristics, such as fibrillar morphology, birefringence upon binding to Congo red and increasedfluorescence intensity with Thioflavine T. Limited resistance to protease K digestion and CD spectroscopyexperiments suggested that the structure of bPrP-UPrD had been changed, and adopted a new, high contentB-sheet conformation during the fibrils formation. Moreover, bPrP-UPrD amyloid fibrils could recruit moresoluble forms into the aggregates. Therefore, the engineered molecules could mimic significant behaviors of PrPse and will be helpful for further understanding the mechanism of conformational conversion process.

  1. 9 CFR 113.65 - Brucella Abortus Vaccine.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Brucella Abortus Vaccine. 113.65... Bacterial Vaccines § 113.65 Brucella Abortus Vaccine. Brucella Abortus Vaccine shall be prepared as a desiccated live culture bacterial vaccine from smooth colonial forms of the Brucella abortus...

  2. Brucella abortus: pathogenicity and gene regulation of virulence

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    Olga Rivas-Solano

    2015-06-01

    Full Text Available Brucella abortus is a zoonotic intracellular facultative pathogen belonging to the subdivision α2 of class Proteobacteria. It causes a worldwide distributed zoonotic disease called brucellosis. The main symptoms are abortion and sterility in cattle, as well as an undulant febrile condition in humans. In endemic regions like Central America, brucellosis has a high socioeconomic impact. A basic research project was recently conducted at the ITCR with the purpose of studying gene regulation of virulence, structure and immunogenicity in B. abortus. The present review was written as part of this project. B. abortus virulence seems to be determined by its ability to invade, survive and replicate inside professional and non-professional phagocytes. It reaches its intracellular replicative niche without the activation of host antimicrobial mechanisms of innate immunity. It also has gene regulation mechanisms for a rapid adaptation to an intracellular environment such as the two-component signal transduction system BvrR/BvrS and the quorum sensing regulator called Vjbr, as well as other transcription factors. All of them integrate a complex gene regulation network.

  3. Recent advances in Brucella abortus vaccines

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    Dorneles, Elaine Maria S.; Sriranganathan, Nammalwar; Andrey P. Lage

    2015-01-01

    Abstract Brucella abortus vaccines play a central role in bovine brucellosis control/eradication programs and have been successfully used worldwide for decades. Strain 19 and RB51 are the approved B. abortus vaccines strains most commonly used to protect cattle against infection and abortion. However, due to some drawbacks shown by these vaccines much effort has been undertaken for the development of new vaccines, safer and more effective, that could also be used in other susceptible species ...

  4. Recent advances in Brucella abortus vaccines

    OpenAIRE

    2015-01-01

    International audience; Brucella abortus vaccines play a central role in bovine brucellosis control/eradication programs and have been successfully used worldwide for decades. Strain 19 and RB51 are the approved B. abortus vaccines strains most commonly used to protect cattle against infection and abortion. However, due to some drawbacks shown by these vaccines much effort has been undertaken for the development of new vaccines, safer and more effective, that could also be used in other susce...

  5. Rapid and specific identification of Brucella abortus using the loop-mediated isothermal amplification (LAMP) assay.

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    Kang, Sung-Il; Her, Moon; Kim, Ji-Yeon; Lee, Jin Ju; Lee, Kichan; Sung, So-Ra; Jung, Suk Chan

    2015-06-01

    A rapid and accurate diagnosis of brucellosis is required to reduce and prevent the spread of disease among animals and the risk of transfer to humans. In this study, a Brucella abortus-specific (Ba) LAMP assay was developed, that had six primers designed from the BruAb2_0168 region of chromosome I. The specificity of this LAMP assay was confirmed with Brucella reference strains, B. abortus vaccine strains, B. abortus isolates and phylogenetically or serologically related strains. The detection limit of target DNA was up to 20 fg/μl within 60 min. The sensitivity of the new LAMP assay was equal to or slightly higher than other PCR based assays. Moreover, this Ba-LAMP assay could specifically amplify all B. abortus biovars compared to previous PCR assays. To our knowledge, this is the first report of specific detection of B. abortus using a LAMP assay. The Ba-LAMP assay can offer a rapid, sensitive and accurate diagnosis of bovine brucellosis in the field.

  6. Recent advances in Brucella abortus vaccines.

    Science.gov (United States)

    Dorneles, Elaine M S; Sriranganathan, Nammalwar; Lage, Andrey P

    2015-07-08

    Brucella abortus vaccines play a central role in bovine brucellosis control/eradication programs and have been successfully used worldwide for decades. Strain 19 and RB51 are the approved B. abortus vaccines strains most commonly used to protect cattle against infection and abortion. However, due to some drawbacks shown by these vaccines much effort has been undertaken for the development of new vaccines, safer and more effective, that could also be used in other susceptible species of animals. In this paper, we present a review of the main aspects of the vaccines that have been used in the brucellosis control over the years and the current research advances in the development of new B. abortus vaccines.

  7. Chlamydophila abortus em animais de produção Chlamydophila abortus in production animals

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    Francielle Gibson da Silva

    2006-02-01

    Full Text Available A Chlamydophila abortus (anteriormente classificada como Chlamydia psittaci sorotipo 1 tem sido descrita em muitos países, associada principalmente com distúrbios reprodutivos em ovinos, bovinos e caprinos. O aborto enzoótico dos ovinos e caprinos e o aborto epizoótico dos bovinos são as doenças mais importantes causadas por esta bactéria. No Brasil, as pesquisas com C. abortus são praticamente inexistentes. O objetivo desta revisão é apresentar informações sobre modificações taxonômicas, ciclo de vida, epidemiologia, patogenia, sinais clínicos e diagnóstico da infecção por C. abortus principalmente em ovinos, bovinos e caprinos.Chlamydophila abortus (previously known as Chlamydia psittaci serovar 1 has been reported in many countries, associated with reproductive disorders in sheep, cattle, and goats. The enzootic abortion of sheep and goats and the epizootic bovine abortion are the most important diseases produced by this bacterium. In Brazil, there is scarce information about C. abortus. The objective of this review is to show information about taxonomic changes, life cycle, epidemiology, pathogenesis, clinical signs and diagnosis of C. abortus in sheep, cattle and goats.

  8. Genetic diversity of Brucella abortus and Brucella melitensis in Kazakhstan using MLVA-16.

    Science.gov (United States)

    Shevtsov, Alexandr; Ramanculov, Erlan; Shevtsova, Elena; Kairzhanova, Alma; Tarlykov, Pavel; Filipenko, Maxim; Dymova, Maya; Abisheva, Gulzada; Jailbekova, Aygul; Kamalova, Dinara; Chsherbakov, Andrei; Tulegenov, Samat; Akhmetova, Assel; Sytnik, Igor; Karibaev, Talgat; Mukanov, Kasim

    2015-08-01

    Brucellosis is an endemic disease in Central Asia characterized by high infection rates in humans and animals. Currently, little is known about the genetic diversity of Brucella spp. circulating in the region, despite the high prevalence of brucellosis. This study aimed to analyze the genetic diversity of Brucella melitensis and Brucella abortus strains circulating in the Republic of Kazakhstan. We genotyped 128 B. melitensis and 124 B. abortus strains collected in regions with the highest prevalence of brucellosis. Genotyping was performed using multi-locus variable-number tandem-repeat analysis (MLVA). Analysis of a subset of 8 loci (MLVA-8) of 128 B. melitensis strains identified genotypes 42 (n=108), 43 (n=2), and 63 (n=19) related to the 'East Mediterranean' group. An MLVA-16 assay sorted 128 B. melitensis strains into 25 different genotypes. Excluding one variable locus, MLVA-15 of B. melitensis was distinct from strains originating in the Mediterranean region; however, 77% of them were identical to strains isolated in China. A minimum spanning tree for B. melitensis using MLVA-15 analysis clustered the local strains together with strains previously collected in China. MLVA-8 analysis of 124 B. abortus strains identified them as genotype 36, suggesting Eurasian distribution of this lineage. Complete MLVA-16 assay analysis clustered the strains into five genotypes, revealing little diversity of B. abortus when compared on the global scale. A minimum spanning tree for B. abortus obtained using MLVA-15 analysis clustered the 2 most prevalent genotypes (n=117) together with strains previously collected in China. Thus, MLVA analysis was used to characterize 252 strains of Brucella collected in Kazakhstan. The analysis revealed genetic homogeneity among the strains. Interestingly, identical MLVA-15 profiles were found in seemingly unrelated outbreaks in China, Turkey, and Kazakhstan. Further analysis is needed for better understanding of the epidemiology of

  9. On the link between cell cycle and infection of the Alphaproteobacterium Brucella abortus

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    Michaël Deghelt

    2014-09-01

    Full Text Available Bacteria of the Brucella genus are responsible for brucellosis, a worldwide zoonosis. These bacteria are known to have a peculiar intracellular trafficking, with a first long and non-proliferative endosomal stage and a second proliferation stage, often associated with its localization of the bacteria in the endoplasmic reticulum (ER. However, the status of the bacterial cell cycle during the non-proliferative phase was still unknown. In a recent study [Nat. Communic. 5:4366], we followed the cell cycle of B. abortus in culture and inside the host cells. In culture, B. abortus initiates the replication of its large chromosome before the small chromosome. The origin and terminator regions of these two chromosomes display distinct localization and dynamics within B. abortus. In HeLa cells and RAW264.7 macrophages, the bacteria in G1 (i.e. before the initiation of chromosomes replication are preferentially found during the endosomal stage of the infection. During this period, growth is also arrested. The cell cycle arrest and resume during the B. abortus trafficking in host cell suggest that like the model Alphaproteobacterium Caulobacter crescentus, these bacteria are able to block their cell cycle at the G1 phase when starvation is sensed.

  10. Genetic characterization of Brucella melitensis and Brucella abortus geographical clusters in Italy.

    Science.gov (United States)

    De Massis, Fabrizio; Garofolo, Giuliano; Cammà, Cesare; Ippoliti, Carla; Candeloro, Luca; Ancora, Massimo; Calistri, Paolo

    2015-01-01

    The genetic diversity of Brucella melitensis and Brucella abortus strains isolated in 199 cattle and sheep from 156 brucellosis outbreaks which occurred in 8 regions of Southern Italy in 2011, was determined using a Multiple-Locus Variable Number of Tandem Repeats Analysis approach. The existence of possible genetic clusters was verified through a hierarchical cluster analysis based on 'single link', which is closely related to the minimum spanning tree. The Hamming weighted distance matrix was adopted in the analysis. All calculations were performed using R and the additional libraries phangorn and Cluster. For a number of clusters, ranging from 2 to 15, the average silhouette width was calculated. The number of clusters adopted was identified according to the maximum average silhouette width. For B. abortus and B. melitensis, 6 and 11 genetic clusters were identified, respectively. Three out of 6 B. abortus clusters included the 96.7% of all B. abortus isolates. Clusters were clearly geographically separated, and this highlighted the known epidemiological links among them. Brucella melitensis genotypes resulted more heterogeneous; the 3 more representative genetic clusters included 79.7% of all B. melitensis isolates. A clear geographical clusterization of genotypes is recognizable only for 1 cluster, whereas the others are more widespread across Southern Italy. The genetic characterization of Brucella strains isolated from animals may be a useful tool to better understand the epidemiology and dissemination patterns of this pathogen through host populations.

  11. Brucella abortus-infected B cells induce osteoclastogenesis.

    Science.gov (United States)

    Pesce Viglietti, Ayelén Ivana; Arriola Benitez, Paula Constanza; Giambartolomei, Guillermo Hernán; Delpino, María Victoria

    2016-09-01

    Brucella abortus is an intracellular bacterium that establishes lifelong infections in livestock and humans although the mechanisms of its chronicity are poorly understood. Activated B cells have long lifespan and B. abortus infection activates B cells. Our results indicate that the direct infection of B cells with B. abortus induced matrix metalloproteinase-9 (MMP-9), receptor activator for NF κB ligand (RANKL), tumor necrosis factor (TNF)-α and interleukin (IL)-6 secretion. In addition, supernatants from B. abortus-infected B cells induced bone marrow-derived monocytes to undergo osteoclastogenesis. Using osteoprotegerin, RANKL's decoy receptor, we determined that RANKL is involved in osteoclastogenesis induced by supernatants from B. abortus-infected B cells. The results presented here shed light on how the interactions of B. abortus with B cells may have a role in the pathogenesis of brucellar osteoarticular disease.

  12. Draft Genome Sequences of Two Brucella abortus Strains Isolated from Cattle and Pig

    OpenAIRE

    2015-01-01

    We report the draft genome sequences of two Brucella abortus strains LMN1 and LMN2 isolated from cattle and pig. The LMN1 and LMN2 have the genome size of 3,395,952 bp and 3,334,792 bp, respectively. In addition to the conserved genes of Brucella, few novel regions showing similarity to the phages were identified in both strains.

  13. Immune response triggered by Brucella abortus following infection or vaccination.

    Science.gov (United States)

    Dorneles, Elaine M S; Teixeira-Carvalho, Andréa; Araújo, Márcio S S; Sriranganathan, Nammalwar; Lage, Andrey P

    2015-07-17

    Brucella abortus live vaccines have been used successfully to control bovine brucellosis worldwide for decades. However, due to some limitations of these live vaccines, efforts are being made for the development of new safer and more effective vaccines that could also be used in other susceptible species. In this context, understanding the protective immune responses triggered by B. abortus is critical for the development of new vaccines. Such understandings will enhance our knowledge of the host/pathogen interactions and enable to develop methods to evaluate potential vaccines and innovative treatments for animals or humans. At present, almost all the knowledge regarding B. abortus specific immunological responses comes from studies in mice. Active participation of macrophages, dendritic cells, IFN-γ producing CD4(+) T-cells and cytotoxic CD8(+) T-cells are vital to overcome the infection. In this review, we discuss the characteristics of the immune responses triggered by vaccination versus infection by B. abortus, in different hosts.

  14. A combined vaccine against Brucella abortus and infectious bovine rhinotracheitis

    OpenAIRE

    2009-01-01

    The present study was undertaken to study the immune response in calves vaccinated with Brucella abortus strain 19, infectious bovine rhinotracheitis (IBR) vaccines in monovalent form and combined vaccine containing both antigen. The seroconversion of monovalent and combined vaccines was tested in seronegative cattle calves. IBR vaccine alone and combination with live Brucella abortus S19 vaccine elicited an anamnestic response on day 60 post booster but started declining from day 90 onwards ...

  15. Seroprevalence of Brucella abortus and Leptospira hardjo in cattle

    OpenAIRE

    2015-01-01

    Aim: The aim was to assess the seroprevalence of B. abortus and Leptospira hardjo in the cattle population of Bihar, this work was carried out. Materials and Methods: Randomly selected 450 cattle from nine districts of Bihar were serologically screened for antibodies against L. hardjo and B. abortus. DAS-ELISA for leptospira and AB-ELISA for brucella were carried out. Based on the results prevalence in each district and the state are reported herewith. Results and Discussion: In this study, i...

  16. Characterization of the Urease Operon of Brucella abortus and Assessment of Its Role in Virulence of the Bacterium▿

    OpenAIRE

    Sangari, Félix J; Seoane, Asunción; Rodríguez, María Cruz; Agüero, Jesús; García Lobo, Juan M.

    2006-01-01

    Most members of the genus Brucella show strong urease activity. However, the role of this enzyme in the pathogenesis of Brucella infections is poorly understood. We isolated several Tn5 insertion mutants deficient in urease activity from Brucella abortus strain 2308. The mutations of most of these mutants mapped to a 5.7-kbp DNA region essential for urease activity. Sequencing of this region, designated ure1, revealed the presence of seven open reading frames corresponding to the urease struc...

  17. Valutazione dell’efficacia del vaccino Brucella abortus ceppo RB51 rispetto al vaccino di referenza Brucella abortus ceppo 19 nel bufalo

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    Massimo Scacchia

    2010-03-01

    Full Text Available Il patrimonio zootecnico della specie bufalina (Bubalus bubalis della regione Campania, è di 250 000 capi, di questi 150 000 allevati in aziende zootecniche della provincia di Caserta. In queste aziende, nel 2007, l’infezione da Brucella abortus ha avuto la prevalenza media, per allevamento, del 20%. Complessivamente, i 2/3 degli allevamenti positivi hanno evidenziato una prevalenza superiore al 10% e, di questi, i 3/4 una prevalenza superiore al 20%. Prendendo il 20% come valore di riferimento, la metà degli allevamenti infetti (22% degli allevamenti casertani ha evidenziato prevalenze inferiori o uguali al 20%, la restante metà (un altro 22% del totale prevalenze comprese tra il 20 e il 56%. In questo contesto epidemiologico è stato adottato un piano di eradicazione della brucellosi che prevedeva l’abbattimento dei capi infetti e la vaccinazione del restante patrimonio bufalino delle zone con più alta incidenza. Per la profilassi vaccinale della brucellosi, il Manual of diagnostic tests and vaccines for terrestrial animals (OIE prevede l’utilizzo del vaccino B. abortus S19 (S19. Purtroppo, l’utilizzo del vaccino negli animali adulti non è privo di possibili effetti indesiderati. Per superare questo aspetto negativo è stato ipotizzato l’impiego del vaccino di B. abortus RB51 (RB51 anche se in letteratura scientifica, sono risultati disponibili pochi dati relativi alla corretta dose vaccinale, all’efficacia e all’innocuità del vaccino nel bufalo. A tale scopo è stato condotto uno studio comparativo tra i due vaccini. Sono state utilizzate 13 femmine di bufalo di 5 mesi di età provenienti da un allevamento ufficialmente indenne da brucellosi. Un gruppo di 5 animali è stato vaccinato due volte, a distanza di un mese, con una dose di RB51 tre volte superiore a quella prevista per i bovini; un secondo gruppo di 5 bufale con S19 rispettando il dosaggio raccomandato per i bovini e un terzo gruppo, di 3 animali di controllo

  18. Detection of Chlamydophila abortus in Sheep (Ovis aries in Mexico

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    Juan M. Jiménez-Estrada

    2008-01-01

    Full Text Available Chlamydophila abortus is one of the pathogens which induce abortion in small ruminants; this pathogen has a tropism for ruminant placenta and causes the disease commonly referred to as Ovine Enzootic Abortion (OEA. In Europe are estimated economic losses of around 20 million pounds a year by OEA. In the American Continent the disease has been reported only in Canada, the United States, Colombia and Chile while in Mexico it is unknown whether OEA is common and it is causing abortions in flocks of sheep from “Estado de Mexico”. The objective of this study was investigating the prevalence of anti-Chlamydophila abortus IgG antibodies and detection of C. abortus DNA in sheep with clinical abort history by mean of ELISA assay (C. abortus ELISA, Institute Pourquier, Montpellier, France and molecular identification of the principal outer membrane protein (POMP 90-91B gene by PCR, respectively. A cross-sectional study was carried out to enroll and random sample of ewes from november 2003 until march 2005. A total of 349 sera and vaginal swabs samples were collected from 35 flocks of sheep from Xalatlaco. The results showed that the seropositive rate was 31.1% (14/45 for healthy and 21.3% (65/304 for sheep with history clinical of abort. In vaginal swabs, the PCR showed 0% (0/45 for healthy animals and 0.65% (2/304 for aborted sheep. Samples from the lungs and liver of the fetus of one of these animals were also positive for C. abortus. In conclusion, these results confirmed that infection with C. abortus is common and is affecting sheep flocks in the Mexican highlands. Therefore, is necessary that the authorities responsible for animal welfare in Mexico (SAGARPA to set up appropriate epidemiological surveillance and control programs to eradicate this disease.

  19. Characterization and Immunogenicity of Outer Membrane Vesicles from Brucella abortus.

    Science.gov (United States)

    Kaur, Gagandeep; Singh, Satparkash; Sunil Kumar, B V; Mahajan, Kanika; Verma, Ramneek

    2016-01-01

    Bovine brucellosis is a worldwide spread zoonotic disease. The objectives of this study were characterization of outer membrane vesicles from B. abortus and to evaluate their immunogenicity in mice. For this purpose, OMVs were derived from B. abortus strain 99 using ultracentrifugation method. Isolated OMVs were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transmission electron microscopy which revealed spherical 20-300 nm structures rich in proteins. OMVs also showed immuno-reactivity with mice antisera in Western blot. Further, indirect ELISA showed specific and high-titer immune responses against the antigens present in OMVs suggesting their potential for a safe acellular vaccine candidate.

  20. Identification of new IS711 insertion sites in Brucella abortus field isolates

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    Moriyón Ignacio

    2011-08-01

    Full Text Available Abstract Background Brucellosis is a zoonosis caused by Brucella spp., a group of highly homogeneous bacteria. The insertion sequence IS711 is characteristic of these bacteria, and occurs in variable numbers and positions, but always constant within a given species. This species-associated polymorphism is used in molecular typing and identification. Field isolates of B. abortus, the most common species infecting cattle, typically carry seven IS711 copies (one truncated. Thus far, IS711 transposition has only been shown in vitro and only for B. ovis and B. pinnipedialis, two species carrying a high number of IS711 copies, but never in other Brucella species, neither in vitro nor in field strains. Results We found several B. abortus strains isolated from milk and aborted fetuses that carried additional IS711 copies in two hitherto undescribed insertion sites: one in an intergenic region near to the 3' end of a putative lactate permease gene and the other interrupting the sequence of a marR transcriptional regulator gene. Interestingly, the second type of insertion was identified in isolates obtained repeatedly from the same herd after successive brucellosis outbreaks, an observation that proves the stability and virulence of the new genotype under natural conditions. Sequence analyses revealed that the new copies probably resulted from the transposition of a single IS711 copy common to all Brucella species sequenced so far. Conclusions Our results show that the replicative transposition of IS711 can occur under field conditions. Therefore, it represents an active mechanism for the emergence of genetic diversity in B. abortus thus contributing to intra-species genetic polymorphism.

  1. MLVA16 typing of Portuguese human and animal Brucella melitensis and Brucella abortus isolates.

    Science.gov (United States)

    Ferreira, Ana Cristina; Chambel, Lélia; Tenreiro, Tania; Cardoso, Regina; Flor, Lídia; Dias, Isabel Travassos; Pacheco, Teresa; Garin-Bastuji, Bruno; Le Flèche, Philippe; Vergnaud, Gilles; Tenreiro, Rogério; de Sá, Maria Inácia Corrêa

    2012-01-01

    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 species but panel 1 and 2A markers displayed higher diversity (HGDI 0.693) in B. abortus compared to B. melitensis isolates (HGDI 0.342). The B. melitensis population belong to the "Americas" (17%) and "East Mediterranean" (83%) groups. No isolate belonged to the "West Mediterranean" group. Eighty-five percent of the human isolates (39 in 46) fit in the "East-Mediterranean" group where a single lineage known as MLVA11 genotype 116 is responsible for the vast majority of Brucella infections in humans. B. abortus isolates formed a consistent group with bv1 and bv3 isolates in different clusters. Four MLVA11 genotypes were observed for the first time in isolates from S. Jorge and Terceira islands from Azores. From the collection of isolates analysed in this study we conclude that MLVA16(Orsay) provided a clear view of Brucella spp. population, confirming epidemiological linkage in outbreak investigations. In particular, it suggests recent and ongoing colonisation of Portugal with one B. melitensis lineage usually associated with East Mediterranean countries.

  2. MLVA16 typing of Portuguese human and animal Brucella melitensis and Brucella abortus isolates.

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    Ana Cristina Ferreira

    Full Text Available 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 species but panel 1 and 2A markers displayed higher diversity (HGDI 0.693 in B. abortus compared to B. melitensis isolates (HGDI 0.342. The B. melitensis population belong to the "Americas" (17% and "East Mediterranean" (83% groups. No isolate belonged to the "West Mediterranean" group. Eighty-five percent of the human isolates (39 in 46 fit in the "East-Mediterranean" group where a single lineage known as MLVA11 genotype 116 is responsible for the vast majority of Brucella infections in humans. B. abortus isolates formed a consistent group with bv1 and bv3 isolates in different clusters. Four MLVA11 genotypes were observed for the first time in isolates from S. Jorge and Terceira islands from Azores. From the collection of isolates analysed in this study we conclude that MLVA16(Orsay provided a clear view of Brucella spp. population, confirming epidemiological linkage in outbreak investigations. In particular, it suggests recent and ongoing colonisation of Portugal with one B. melitensis lineage usually associated with East Mediterranean countries.

  3. Reporte de primer caso humano de aislamiento y tipificación de Brucella abortus RB 51: first report in Chile Isolation and identification of Brucella abortus RB 51 in human

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    M. Villarroel

    2000-01-01

    Full Text Available En el Laboratorio de Microbiología del Hospital Base de Osorno, se aisló una cepa de Brucella sp, que posteriormente se tipificó en el Laboratorio Regional de Diagnósticos del Servicio Agrícola y Ganadero, SAG Osorno. Esta cepa fue aislada de un hemocultivo realizado a un paciente con sintomatología clínica compatible con Brucelosis. El resultado de la tipificación fue Brucella abortus RB 51, cepa vaccinal que se presumía apatógena para el hombre. Este es el primer reporte a nivel mundial de un aislamiento de B. abortus RB 51 en humanosA Brucella strain was isolated from a haemoculture of a clinically ill veterinarian practitioner, at the Laboratory of Microbiology, Hospital Base Osorno, Chile. This strain was identified as Brucella abortus RB 51 at Laboratorio de Diagnósticos from the Servicio Agrícola y Ganadero (SAG, Osorno, based mainly in the following characteristics: resistence to Rifampicin, aerobic growth and the production of only rough colonies. The strain RB 51 was confirmed by PCR analysis

  4. Circulating Strains of Brucella abortus in Cattle in Santo Domingo De Los Tsáchilas Province - Ecuador.

    Science.gov (United States)

    Rodríguez-Hidalgo, Richar Ivan; Contreras-Zamora, Javier; Benitez Ortiz, Washington; Guerrero-Viracocha, Karina; Salcan-Guaman, Holger; Minda, Elizabeth; Ron Garrido, Lenin

    2015-01-01

    The Province of Santo Domingo de los Tsáchilas in Ecuador represents the largest informal cattle market. Because of its strategic position, cattle movement is very high and therefore we selected this region, to determine the strain variation of Brucella sp. Part of the study aimed at the isolation, biotyping, and genotyping of Brucella species from milk and supra-mammary lymph nodes of sero-positive bovines, using selective Farrell medium, biochemical assays, and IS711-PCR, AMOS-PCR, and HOOF-Prints techniques. In total, 656 animals from 12 sero-positive dairy herds and from the provincial slaughterhouse were diagnosed by Rose Bengal and Wright's Slow Agglutination test with EDTA. Amongst these animals, 50 animals were sero-positive for brucellosis. Twenty-five lymph nodes and 25 milk samples from each group of positive reactors were transferred to culture medium. Isolation was possible from 4 (16%) lymph nodes and 9 (36%) milk samples; out of these, 10 isolates were diagnosed as Brucella sp. All four isolates of lymphatic tissue corresponded to Brucella abortus biotype 1, confirmed as field strains by molecular analysis. Milk isolations, showed biochemically a more dispersed pattern in which B. abortus biotypes 1 and 4 were found; yet four samples gave a pattern similar to B. abortus biotype 2; however, only biotypes 1 and 4 were confirmed by molecular analysis. The concordance between biochemical and molecular diagnostic tests reached 76.9%.

  5. Seroprevalence of Brucella abortus and Leptospira hardjo in cattle

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    S. Jegaveera Pandian

    2015-02-01

    Full Text Available Aim: The aim was to assess the seroprevalence of B. abortus and Leptospira hardjo in the cattle population of Bihar, this work was carried out. Materials and Methods: Randomly selected 450 cattle from nine districts of Bihar were serologically screened for antibodies against L. hardjo and B. abortus. DAS-ELISA for leptospira and AB-ELISA for brucella were carried out. Based on the results prevalence in each district and the state are reported herewith. Results: In this study, it was found that the seroprevalence of L. hardjo was 9.11% and that of B. abortus was 12.2% in Bihar. Indigenous cattle were found to be less susceptible to leptospirosis and brucellosis even though they accounted for 83.11% of the study population. Conclusion: Although there was no acute disease, antibodies detected against L. hardjo and B. abortus in the cattle population indicated the presence of chronic and subclinical infection, which could challenge the fertility of the animals.

  6. Brucella abortus RB51 in milk of vaccinated adult cattle.

    Science.gov (United States)

    Miranda, Karina Leite; Poester, Fernando Padilla; Dorneles, Elaine Maria Seles; Resende, Thiago Magalhães; Vaz, Adil Knackfuss; Ferraz, Sandra Maria; Lage, Andrey Pereira

    2016-08-01

    The aim of this study was to evaluate the shedding of Brucella abortus in the milk of cows vaccinated with a full dose of RB51 during lactation. Eighteen cows, nine previously vaccinated with S19 as calves and nine non-vaccinated, were immunized subcutaneously with 1.3×10(10)CFU of B. abortus RB51, 30-60days after parturition. Milk samples from all animals were collected daily until day 7, and at weekly interval for the next 9 weeks after vaccination. To evaluate the shedding of B. abortus, milk samples were submitted for culture and PCR. No B. abortus was isolated from any sample tested. Only one sample, collected on first day after vaccination from a cow previously vaccinated, was faintly positive in the PCR. In conclusion, the public health hazard associated with milk consumption from cows vaccinated with RB51 in post-partum is very low, despite vaccination with the full dose and regardless of previous S19 vaccination.

  7. Endocarditis por Brucella abortus: Reporte del primer caso en C.R Brucella abortus Endocarditis

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    Manuel Antonio Villalobos-Zúñiga

    2011-09-01

    Full Text Available Paciente masculino de 36 años de edad, proveniente de la zona rural de Costa Rica, con un cuadro clínico de 8 meses de evolución de fiebre, mialgias, artralgias, pérdida de peso y lumbalgia; referido por la detección de un soplo de insuficiencia aórtica. El ecocardiograma reveló endocarditis de la válvula aórtica, y se obtuvieron 4 hemocultivos positivos por Brucella abortus biotipo 3, con serologías negativas por brucelosis. Se inició tratamiento con antibióticos y luego se le realizó un reemplazo valvular aórtico; 4 meses después ingresó con dolor torácico que se atribuyó a una oclusión de la arteria descendente anterior, demostrada angiográficamente, por posible embolismo. En la actualidad cursa clínicamente estable con manejo médico para su cardiopatía, sin recaída infecciosa.The case of a 36-year-old patient from a rural area is presented. He came with an 8 month history of fever, myalgias, arthralgias, weight loss and lower back pain; who also had an aortic insufficiency murmur detected. The diagnosis of aortic valve endocarditis was made by echocardiography, and had 4 positive blood cultures for Brucella abortus biotype 3, and negative serologic test for brucellosis. He was started on antibiotics and later on underwent aortic valve replacement, with a late coronary cardioembolism as a complication.

  8. High-resolution melt PCR analysis for rapid identification of Chlamydia abortus live vaccine strain 1B among C. abortus strains and field isolates.

    Science.gov (United States)

    Vorimore, Fabien; Cavanna, Noémie; Vicari, Nadia; Magnino, Simone; Willems, Hermann; Rodolakis, Annie; Siarkou, Victoria I; Laroucau, Karine

    2012-09-01

    We describe a novel high-resolution melt assay that clearly differentiates Chlamydia abortus live vaccine strain 1B from field C. abortus strains and field wild-type isolates based on previously described single nucleotide polymorphisms. This modern genotyping technique is inexpensive, easy to use, and less time-consuming than PCR-RFLP.

  9. Identification and characterization of Chlamydia abortus isolates from yaks in Qinghai, China.

    Science.gov (United States)

    Li, Zhaocai; Cao, Xiaoan; Fu, Baoquan; Chao, Yilin; Cai, Jinshan; Zhou, Jizhang

    2015-01-01

    Recently, the yak population has exhibited reproductive disorders, which are considered to be associated with Chlamydia abortus (C. abortus) in Qinghai, China. In this study, a total of 9 aborted fetuses (each from a different herd) and 126 vaginal swab samples from the 9 herds were collected and analyzed. C. abortus DNA was detected from all of the 9 aborted fetuses and 30 of the 126 vaginal swab samples (23.81%) from yak cows in the selected herds. Four C. abortus strains were isolated from embryonated egg yolk sacs inoculated with foetal organ suspensions. The isolated C. abortus strains were further identified, which showed identical restriction profiles with the C. abortus reference strain using AluI restriction enzyme in the RFLP test. Moreover, the isolated C. abortus strains and C. abortus-positive vaginal swab samples were genotyped by multiple loci variable number tandem repeat analysis and all belonged to the genotype 2 group. These findings suggested that C. abortus played a substantial role in yak abortion in Qinghai, China.

  10. Brucella abortus Cell Cycle and Infection Are Coordinated.

    Science.gov (United States)

    De Bolle, Xavier; Crosson, Sean; Matroule, Jean-Yves; Letesson, Jean-Jacques

    2015-12-01

    Brucellae are facultative intracellular pathogens. The recent development of methods and genetically engineered strains allowed the description of cell-cycle progression of Brucella abortus, including unipolar growth and the ordered initiation of chromosomal replication. B. abortus cell-cycle progression is coordinated with intracellular trafficking in the endosomal compartments. Bacteria are first blocked at the G1 stage, growth and chromosome replication being resumed shortly before reaching the intracellular proliferation compartment. The control mechanisms of cell cycle are similar to those reported for the bacterium Caulobacter crescentus, and they are crucial for survival in the host cell. The development of single-cell analyses could also be applied to other bacterial pathogens to investigate their cell-cycle progression during infection.

  11. An ecological perspective on the changing face of Brucella abortus in the western United States

    Science.gov (United States)

    Cross, Paul C.; Maichak, Eric J.; Brennan, Angela; Scurlock, Brandon M.; Henningsen, John C.; Luikart, Gordon

    2013-01-01

    After a hiatus during the 1990s, outbreaks of Brucella abortus in cattle are occurring more frequently in some of the western states of the United States, namely, Montana, Wyoming and Idaho. This increase is coincident with increasing brucellosis seroprevalence in elk (Cervus elaphus), which is correlated with elk density. Vaccines are a seductive solution, but their use in wildlife systems remains limited by logistical, financial, and scientific constraints. Cattle vaccination is ongoing in the region. Livestock regulations, however, tend to be based on serological tests that test for previous exposure and available vaccines do not protect against seroconversion. The authors review recent ecological studies of brucellosis, with particular emphasis on the Greater Yellowstone Area, and highlight the management options and implications of this work, including the potential utility of habitat modifications and targeted hunts, as well as scavengers and predators. Finally, the authors discuss future research directions that will help us to understand and manage brucellosis in wildlife.

  12. Immunization of Mice with Recombinant Brucella abortus Organic Hydroperoxide Resistance (Ohr) Protein Protects Against a Virulent Brucella abortus 544 Infection.

    Science.gov (United States)

    Hop, Huynh Tan; Reyes, Alisha Wehdnesday Bernardo; Simborio, Hannah Leah Tadeja; Arayan, Lauren Togonon; Min, Won Gi; Lee, Hu Jang; Lee, Jin Ju; Chang, Hong Hee; Kim, Suk

    2016-01-01

    In this study, the Brucella abortus ohr gene coding for an organic hydroperoxide resistance protein (Ohr) was cloned into a maltose fusion protein expression system (pMAL), inserted into Escherichia coli, and purified, and its immunogenicity was evaluated by western blot analysis using Brucella-positive mouse sera. The purified recombinant Ohr (rOhr) was treated with adjuvant and injected intraperitoneally into BALB/c mice. A protective immune response analysis revealed that rOhr induced a significant increase in both the IgG1 and IgG2a titers, and IgG2a reached a higher level than IgG1 after the second and third immunizations. Additionally, immunization with rOhr induced high production of IFN-γ as well as proinflammatory cytokines such as TNF, MCP-1, IL-12p70, and IL-6, but a lesser amount of IL-10, suggesting that rOhr predominantly elicited a cell-mediated immune response. In addition, immunization with rOhr caused a significantly higher degree of protection against a virulent B. abortus infection compared with a positive control group consisting of mice immunized with maltose-binding protein. These findings showed that B. abortus rOhr was able to induce both humoral and cell-mediated immunity in mice, which suggested that this recombinant protein could be a potential vaccine candidate for animal brucellosis.

  13. Vaccination of elk (Cervus canadensis with Brucella abortus strain RB51 overexpressing superoxide dismutase and glycosyltransferase genes does not induce adequate protection against experimental Brucella abortus challenge

    Directory of Open Access Journals (Sweden)

    Pauline eNol

    2016-02-01

    Full Text Available In recent years, elk (Cervus canadensis have been implicated as the source of Brucella abortus infection for numerous cattle herds in the Greater Yellowstone Area. In the face of environmental and ecological changes on the landscape, the range of infected elk is expanding. Consequently, the development of effective disease management strategies for wild elk herds is of utmost importance, not only for the prevention of reintroduction of brucellosis to cattle, but also for the overall health of the Greater Yellowstone Area elk populations. In two studies, we evaluated the efficacy of B. abortus strain RB51 over-expressing superoxide dismutase and glycosytransferase for protecting elk from infection and disease caused by B. abortus after experimental infection with a virulent B. abortus strain. Our data indicate that the recombinant vaccine does not protect elk against brucellosis. Further work is needed for development of an effective brucellosis vaccine for use in elk

  14. Vaccination of Elk (Cervus canadensis) with Brucella abortus Strain RB51 Overexpressing Superoxide Dismutase and Glycosyltransferase Genes Does Not Induce Adequate Protection against Experimental Brucella abortus Challenge.

    Science.gov (United States)

    Nol, Pauline; Olsen, Steven C; Rhyan, Jack C; Sriranganathan, Nammalwar; McCollum, Matthew P; Hennager, Steven G; Pavuk, Alana A; Sprino, Phillip J; Boyle, Stephen M; Berrier, Randall J; Salman, Mo D

    2016-01-01

    In recent years, elk (Cervus canadensis) have been implicated as the source of Brucella abortus infection for numerous cattle herds in the Greater Yellowstone Area. In the face of environmental and ecological changes on the landscape, the range of infected elk is expanding. Consequently, the development of effective disease management strategies for wild elk herds is of utmost importance, not only for the prevention of reintroduction of brucellosis to cattle, but also for the overall health of the Greater Yellowstone Area elk populations. In two studies, we evaluated the efficacy of B. abortus strain RB51 over-expressing superoxide dismutase and glycosyltransferase for protecting elk from infection and disease caused by B. abortus after experimental infection with a virulent B. abortus strain. Our data indicate that the recombinant vaccine does not protect elk against brucellosis. Further, work is needed for development of an effective brucellosis vaccine for use in elk.

  15. Infecção em cão por Brucella abortus: relato de caso Brucella abortus infection in dog: case report

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    J. Megid

    2007-12-01

    Full Text Available Brucella abortus infection is reported in a dog from a rural area that presented at clinical evaluation left testicular enlargement and right testicular decrease. Serum resulted negative to rapid agglutination test and agar gel immunodifusion with Brucella ovis antigen but positive to buffered plate agglutination test, tube agglutination test and 2- Mercapthoetanol with B. abortus antigen. Brucella isolation was negative in blood, testicular material, semen and urine. Brucella DNA was detected in PCR from urine and blood.

  16. Circulating strains of Brucella abortus in cattle in the province of Santo Domingo de los Tsáchilas - Ecuador.

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    Richar Ivan Rodríguez Hidalgo

    2015-03-01

    Full Text Available In Ecuador, the Province of Santo Domingo de los Tsáchilas represents the largest informal market of livestock due to its strategic position in the country; thus, given the high mobility of cattle in this region, the aim of this study was to determine the strain variation of Brucella sp.. Part of the study was the isolation, biotyping and genotyping of Brucella species isolated from milk and supra-mammary lymph nodes of sero-positive bovines. Protocols used for isolation, biotyping and genotyping of Brucella species were selective Farrell medium, biochemical assays and IS711-PCR, AMOS-PCR and HOOF-Prints techniques, respectively. In total, 656 animals from sero-positive dairy herds and from the slaughterhouse of the province were diagnosed by Rose Bengal and Wright’s Slow Agglutination test with EDTA. From these animals, 50 animals were found sero-positive for brucellosis. Twenty-five lymph nodes and 25 milk samples from each group of positive reactors were cultured and growth. Isolation was possible in 4 (16% and 9 (36%, respectively; and of these, 10 isolates were diagnosed as Brucella sp. All 4 isolates of lymphatic tissue corresponded to Brucella abortus biotype 1, confirmed as field strains by molecular analysis. Milk isolations, biochemically showed a more dispersed pattern in which B. abortus biotypes 1 and 4 were found; yet four samples gave a pattern similar to B. abortus biotype 2; however, only biotypes 1 and 4 were confirmed by molecular analysis. The concordance between biochemical and molecular diagnostic tests reached 76.9%.

  17. A dual-targeting approach to inhibit Brucella abortus replication in human cells

    Science.gov (United States)

    Czyż, Daniel M.; Jain-Gupta, Neeta; Shuman, Howard A.; Crosson, Sean

    2016-01-01

    Brucella abortus is an intracellular bacterial pathogen and an etiological agent of the zoonotic disease known as brucellosis. Brucellosis can be challenging to treat with conventional antibiotic therapies and, in some cases, may develop into a debilitating and life-threatening chronic illness. We used multiple independent assays of in vitro metabolism and intracellular replication to screen a library of 480 known bioactive compounds for novel B. abortus anti-infectives. Eighteen non-cytotoxic compounds specifically inhibited B. abortus replication in the intracellular niche, which suggests these molecules function by targeting host cell processes. Twenty-six compounds inhibited B. abortus metabolism in axenic culture, thirteen of which are non-cytotoxic to human host cells and attenuate B. abortus replication in the intracellular niche. The most potent non-cytotoxic inhibitors of intracellular replication reduce B. abortus metabolism in axenic culture and perturb features of mammalian cellular biology including mitochondrial function and receptor tyrosine kinase signaling. The efficacy of these molecules as inhibitors of B. abortus replication in the intracellular niche suggests “dual-target” compounds that coordinately perturb host and pathogen are promising candidates for development of improved therapeutics for intracellular infections. PMID:27767061

  18. Caracterização epidemiológica e fatores de risco associados à infecção por Chlamydophila abortus em ovinos deslanados do semiárido brasileiro Epidemiological characterization and risk factors associated with Chlamydophila abortus infection in sheep in Brazilian semiarid

    Directory of Open Access Journals (Sweden)

    Areano E.M. Farias

    2013-03-01

    Full Text Available O objetivo do presente trabalho foi determinar as prevalências de propriedades positivas e animais soropositivos para Chlamydophila abortus em ovinos deslanados da região semiárida do Nordeste do Brasil, bem como identificar fatores de risco. Foram colhidas amostras de sangue de 476 ovinos procedentes de 72 propriedades em 14 municípios na mesorregião do Sertão, Estado da Paraíba. Para o diagnóstico sorológico da infecção por Chlamydophila abortus foi utilizada a reação de fixação de complemento (RFC. Uma propriedade foi considerada positiva quando apresentou pelo menos um animal soropositivo. Das 72 propriedades usadas 38 (52,8% apresentaram pelo menos um animal soropositivo, e dos 476 animais 94 (19,7% foram soropositivos. Participação em exposições (odds ratio =4,31; IC 95% =1,80-10,35; p=0,011 foi identificada como fator de risco. Sugere-se que a infecção por Chlamydophila abortus encontra-se disseminada em ovinos da região, e baseando-se na análise de fatores de risco, recomenda-se o controle sanitário nas exposições de animais.The aim of this investigation was to determine the flock-level and animal-level prevalences of Chlamydophila abortus in sheep from the semiarid region of Northeastern Brazil, as well as to identify risk factors. Blood samples were collected from 476 sheep of 72 flocks in 14 counties in the Sertão mesoregion, state of Paraíba. For the serological diagnosis of Chlamydophila abortus infection the complement fixation test (FC was carried out. A flock was positive when presented at least one seropositive animal. From the 72 flocks, 38 (52.8% presented at least one seropositive sheep, and 94 (19.7% of the 476 animals were seropositive. Participation in animal expositions (odds ratio= 4.31; 95% CI= 1.80-10.35; p=0.011 was identified as risk factor. It is suggested that C. abortus infection is spread in sheep of the region, and based on the risk factor analysis sanitary control in animal

  19. Chlamydia abortus in Cows Oviducts, Occasional Event or Causal Connection?

    Science.gov (United States)

    Appino, S; Vincenti, L; Rota, A; Pellegrini, S; Chieppa, M N; Cadoni, V; Pregel, P

    2015-06-01

    Fifty-seven genital tracts of regularly slaughtered culled Piedmontese cows, aged 7.4 ± 4.3 years (mean ± SD), range: 2.6-15.6 years, were grossly and microscopically examined. DNA extracted from oviducts was subjected to PCR to evaluate the presence of Chlamydia spp. The 15 PCR-positive oviducts were subjected to Sanger sequencing and showed the presence of Chamydia abortus, with an identity range between 99 and 100%. Nine of the PCR-positive samples belonged to the 24 animals with a normal macroscopic appearance of the whole genital tract (percentage of positive oviducts in normal genital tracts 9/24 = 37.5%), while six belonged to the 33 genital tracts with lesions in one or more organs (percentage of positive oviducts in pathological genital tracts 6/33 = 18.1%); of these, a single animal had salpingitis. The detection of C. abortus in bovine oviducts is of particular interest because it has never been previously investigated or reported.

  20. Detection of Brucella abortus in Chiredzi district in Zimbabwe

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    Calvin Gomo

    2012-02-01

    Full Text Available Brucellosis is an endemic disease in Zimbabwe caused by the genus Brucella. Brucella seroprevalence was recently reported to be high in the wildlife-livestock interface in the Chiredzi district and the neighbouring Gonarezhou National Park (GNP in Zimbabwe, and higher amongst communal cattle with an abortion history and access to grazing in GNP than amongst communal cattle with no abortion history or access to grazing in GNP. The aim of this study was to investigate Brucella species in brucellosis seropositive cattle in the Chiredzi district with access to GNP using isolation and identification. Isolation of Brucella species from whole blood (n = 18 and milk samples (n = 10 from seropositive animals with an abortion history was based on the rose Bengal test (RBT and enzyme-linked immunoassays (enzyme- linked immunosorbent assay [ELISA]; indirect ELISA and complement ELISA, using microbiology and polymerase chain reaction (PCR methods. Brucella abortus was cultured and identified from blood and milk collected from seropositive cows in both communal areas. The Brucella-specific 16-23S intergenic spacer (ITS PCR and multiplex AMOS-PCR assays verified the identification of the cultures. Our results confirmed that B. abortus is present in cattle on communal farms in the Chiredzi district in Zimbabwe and might cause cattle abortions. The need for implementing control measures and raising public awareness on zoonotic transmission of brucellosis are recommended.

  1. Immunogenicity and protective effect of recombinant Brucella abortus Ndk (rNdk) against a virulent strain B. abortus 544 infection in BALB/c mice.

    Science.gov (United States)

    Hop, Huynh Tan; Simborio, Hannah Leah; Reyes, Alisha Wehdnesday Bernardo; Arayan, Lauren Togonon; Min, WonGi; Lee, Hu Jang; Kim, Dong Hee; Chang, Hong Hee; Kim, Suk

    2015-02-01

    In this study, we particularly evaluated the protective effect of recombinant protein encoded by Brucella abortus 544 ndk (nucleoside diphosphate kinase) gene against B. abortus infection in the BALB/c mice. Cloning and expression of B. abortus Ndk was accomplished by PCR amplification into a pMAL expression system, and purification of a recombinant Ndk (rNdk). As for the determination of IgG responses, rNdk induced vigorous IgG production, especially higher in IgG2a compared to IgG1 with titers of 5.2 and 4.8, respectively, whereas titers of these in mice immunized with MBP were 2.4 of IgG2a and 2.6 of IgG1. The analysis of cytokine has revealed that rNdk can strongly induce production of IFN-γ as well as proinflammatory cytokines (TNF, MCP1 and IL-6) but not much IL-10, suggesting rNdk elicited predominantly cell-mediated immune responses. Furthermore, the spleen proliferation and bacterial burden in the spleen of rNdk immunized mice were significantly lower than those of MBP-immunized mice against virulent B. abortus challenge (P abortus might be a useful candidate for subunit vaccine for brucellosis in animals.

  2. LAPORAN KASUS ABORTUS IMINENS JUNI 2015 FAKTOR RESIKO, PATOGENESIS, DAN PENATALAKSANAAN

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    Kiki Sanjaya Dharma

    2015-08-01

    Full Text Available ABSTRACT Introduction: One of the most complications in pregnancy it was a bleeding , that could happen to any the gestational age .Young in pregnancy , often connected with abortus , miscarriage , and loss of early pregnancy .Abortus is defined as the threat or spending the products of conception before a fetus can live outside the utery. Constitutes gestational age is less than 20 weeks or fetus weight less than 500 grams .Abortus imminens is pervaginam bleeding in pregnancy before 20 weeks without accompanied the release of the products of conception and dilatation of the uterus. Cases: Female patient , 31 years , the balinese ethnic coming to the emergency room the hospital with complaints pervaginam bleeding. Said to be initially were bleeding spots that its color red .Then there are black coloured bloodlump, there is no tissue. Kata Kunci :Abortus imminens, pregnancy, bleeding.

  3. Confirmation of Chlamydophila abortus in infected cell culture using Indirect Immunofluorescence technique

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    Krishnan Nair G

    Full Text Available Chlamydophila abortus (C. abortus is an important abortifacient agent in bovines and ovines. Clinical diagnosis of the disease is often difficult. An early diagnosis can be achieved based on direct demonstration of the organism in clinical material and through the cultural recovery of the organism in embryonated chicken egg. For confirmatory diagnosis antigen detection methods or serological techniques can be adopted. The present study is aimed at the confirmatory diagnosis of C. abortus infection by indirect immunofluorescence technique following the isolation of the organism in cell culture. Specific apple green fluorescing inclusions of C. abortus in McCoy cell lines was detected from 72 h to 96 h post infection employing anti-chlamydial group specific monoclonal antibodies. Thus, a confirmatory diagnosis of the infection was possible with this study. [Vet. World 2011; 4(10.000: 473-474

  4. A dual-targeting approach to inhibit Brucella abortus replication in human cells

    OpenAIRE

    2016-01-01

    Brucella abortus is an intracellular bacterial pathogen and an etiological agent of the zoonotic disease known as brucellosis. Brucellosis can be challenging to treat with conventional antibiotic therapies and, in some cases, may develop into a debilitating and life-threatening chronic illness. We used multiple independent assays of in vitro metabolism and intracellular replication to screen a library of 480 known bioactive compounds for novel B. abortus anti-infectives. Eighteen non-cytotoxi...

  5. Survival of Brucella abortus in milk fermented with a yoghurt starter culture.

    Science.gov (United States)

    Zúñiga Estrada, Armida; Mota de la Garza, Lydia; Sánchez Mendoza, Miroslava; Santos López, Eva María; Filardo Kerstupp, Santiago; López Merino, Ahidé

    2005-01-01

    In countries such as Mexico, brucellosis is still an important public health problem due to the consumption of non-pasteurized milk and dairy products, contaminated with Brucella spp. The aim of this study was to look into the survival of Brucella abortus during fermentation of milk with a yoghurt starter culture and storage at refrigeration temperature. Sterile skim milk was inoculated with B. abortus at two concentrations, 10(5) and 10(8) CFU/ml simultaneously with a yoghurt starter culture of lactic acid bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii subspecie bulgaricus). Inoculated flasks were incubated at 42 degrees C, followed by refrigeration at 4 degrees C. Samples were taken during fermentation and during storage and viable count of B. abortus and lactic acid bacteria and pH were determined. Results showed that after 10 days of storage at 4 degrees C, B. abortus was recovered in fermented milk at a level of 10(5) CFU/ml, despite the low pH below 4.0. Therefore B. abortus is able to survive in fermented milk. This finding may imply that non-pasteurized fermented milk contaminated with Brucella abortus could be a means of transmission of these bacteria.

  6. Risks of Brucella abortus spillover in the Greater Yellowstone area.

    Science.gov (United States)

    Schumaker, B

    2013-04-01

    Recurrent spillover of Brucella abortus from wildlife reservoirs to domestic cattle in the Greater Yellowstone Area (GYA) has prevented the United States from completely eradicating bovine brucellosis. Risks to cattle are a function of the size and location of wildlife and livestock populations, the degree and nature of spatio-temporal interactions between the various hosts, the level of disease in wildlife, and the susceptibility of livestock herds. While the brucellosis prevalence in wild, free-ranging GYA bison (Bison bison) is high, current management actions have successfully limited contact between bison and cattle. Under current management practices, the risks to cattle in the GYA are predominantly from wild elk (Cervus elaphus). Intra- and inter-species transmission events, while uncommon, are nevertheless crucial for the maintenance of brucellosis in the GYA. Future management actions should focus on decreasing elk herd densities and group sizes and on understanding the behavioural and environmental drivers that result in co-mingling that makes transmission possible.

  7. Diversification and Distribution of Ruminant Chlamydia abortus Clones Assessed by MLST and MLVA.

    Science.gov (United States)

    Siarkou, Victoria I; Vorimore, Fabien; Vicari, Nadia; Magnino, Simone; Rodolakis, Annie; Pannekoek, Yvonne; Sachse, Konrad; Longbottom, David; Laroucau, Karine

    2015-01-01

    Chlamydia abortus, an obligate intracellular bacterium, is the most common infectious cause of abortion in small ruminants worldwide and has zoonotic potential. We applied multilocus sequence typing (MLST) together with multiple-locus variable-number tandem repeat analysis (MLVA) to genotype 94 ruminant C. abortus strains, field isolates and samples collected from 1950 to 2011 in diverse geographic locations, with the aim of delineating C. abortus lineages and clones. MLST revealed the previously identified sequence types (STs) ST19, ST25, ST29 and ST30, plus ST86, a recently-assigned type on the Chlamydiales MLST website and ST87, a novel type harbouring the hemN_21 allele, whereas MLVA recognized seven types (MT1 to MT7). Minimum-spanning-tree analysis suggested that all STs but one (ST30) belonged to a single clonal complex, possibly reflecting the short evolutionary timescale over which the predicted ancestor (ST19) has diversified into three single-locus variants (ST86, ST87 and ST29) and further, through ST86 diversification, into one double-locus variant (ST25). ST descendants have probably arisen through a point mutation evolution mode. Interestingly, MLVA showed that in the ST19 population there was a greater genetic diversity than in other STs, most of which exhibited the same MT over time and geographical distribution. However, the evolutionary pathways of C. abortus STs seem to be diverse across geographic distances with individual STs restricted to particular geographic locations. The ST30 singleton clone displaying geographic specificity and represented by the Greek strains LLG and POS was effectively distinguished from the clonal complex lineage, supporting the notion that possibly two separate host adaptations and hence independent bottlenecks of C. abortus have occurred through time. The combination of MLST and MLVA assays provides an additional level of C. abortus discrimination and may prove useful for the investigation and surveillance of

  8. Diversification and Distribution of Ruminant Chlamydia abortus Clones Assessed by MLST and MLVA.

    Directory of Open Access Journals (Sweden)

    Victoria I Siarkou

    Full Text Available Chlamydia abortus, an obligate intracellular bacterium, is the most common infectious cause of abortion in small ruminants worldwide and has zoonotic potential. We applied multilocus sequence typing (MLST together with multiple-locus variable-number tandem repeat analysis (MLVA to genotype 94 ruminant C. abortus strains, field isolates and samples collected from 1950 to 2011 in diverse geographic locations, with the aim of delineating C. abortus lineages and clones. MLST revealed the previously identified sequence types (STs ST19, ST25, ST29 and ST30, plus ST86, a recently-assigned type on the Chlamydiales MLST website and ST87, a novel type harbouring the hemN_21 allele, whereas MLVA recognized seven types (MT1 to MT7. Minimum-spanning-tree analysis suggested that all STs but one (ST30 belonged to a single clonal complex, possibly reflecting the short evolutionary timescale over which the predicted ancestor (ST19 has diversified into three single-locus variants (ST86, ST87 and ST29 and further, through ST86 diversification, into one double-locus variant (ST25. ST descendants have probably arisen through a point mutation evolution mode. Interestingly, MLVA showed that in the ST19 population there was a greater genetic diversity than in other STs, most of which exhibited the same MT over time and geographical distribution. However, the evolutionary pathways of C. abortus STs seem to be diverse across geographic distances with individual STs restricted to particular geographic locations. The ST30 singleton clone displaying geographic specificity and represented by the Greek strains LLG and POS was effectively distinguished from the clonal complex lineage, supporting the notion that possibly two separate host adaptations and hence independent bottlenecks of C. abortus have occurred through time. The combination of MLST and MLVA assays provides an additional level of C. abortus discrimination and may prove useful for the investigation and

  9. Vaccination of elk (Cervus canadensis) with Brucella abortus strain RB51 overexpressing superoxide dismutase and glycosyltransferase genes does not induce adequate protection against experimental brucella abortus challenge

    Science.gov (United States)

    In recent years, elk (Cervus canadensis) have been implicated as the source of Brucella abortus infection for numerous cattle herds in the Greater Yellowstone Area (GYA). In the face of environmental and ecological changes on the landscape, the range of infected elk is expanding. Consequently, the d...

  10. Literatuuronderzoek naar gegevens betreffende de betekenis van een aantal verwekkers van zoonosen in verband met de vleesconsumptie. VIII: Brucella abortus

    NARCIS (Netherlands)

    Bos; J.M.; Engel; H.W.B.; Groothuis; D.G.; Knapen; F. van; Weiss; J.W.

    1986-01-01

    Brucellose bij de mens heeft meestal een weinig specifiek, griepachtig beeld. In Nederland is de voornaamste verwekker Brucella abortus, waar toe het literatuuronderzoek zich dan ook beperkt. Bij dieren is het rund de voornaamste gastheer en leidt een infectie bij drachtige dieren tot abortus.

  11. Complete Genome Sequence of Brucella abortus SKN 13 Isolated from Placenta of Aborted Cattle in Gujarat, India

    Science.gov (United States)

    Chauhan, H. C.; Chandel, B. S.; Patel, Kirit B.; Patel, A. C.; Shrimali, M. D.; Patel, S. S.; Bhagat, A. G.; Rajgor, Manish; Patel, Mitul A.; Patel, Maulik; Kala, Jitendra; Patel, Bhumika

    2016-01-01

    Brucella abortus is generally known to cause brucellosis in cattle and buffalo. Here, we report the draft genome sequence of Brucella abortus SKN 13, isolated from aborted cattle placenta in the area of Gujarat, India, providing precious resources for comparative genomic analyses of Brucella field strains. PMID:27789633

  12. Complete Genome Sequence of Brucella abortus SKN 13 Isolated from Placenta of Aborted Cattle in Gujarat, India

    OpenAIRE

    2016-01-01

    Brucella abortus is generally known to cause brucellosis in cattle and buffalo. Here, we report the draft genome sequence of Brucella abortus SKN 13, isolated from aborted cattle placenta in the area of Gujarat, India, providing precious resources for comparative genomic analyses of Brucella field strains.

  13. Characterization of the immunogenicity and pathogenicity of malate dehydrogenase in Brucella abortus.

    Science.gov (United States)

    Han, Xiangan; Tong, Yongliang; Tian, Mingxing; Sun, Xiaoqing; Wang, Shaohui; Ding, Chan; Yu, Shengqing

    2014-07-01

    Brucella abortus is a gram-negative, facultative intracellular pathogen that causes brucellosis, a chronic zoonotic disease resulting in abortion in pregnant cattle and undulant fever in humans. Malate dehydrogenase (MDH), a key enzyme in the tricarboxylic acid cycle, plays important metabolic roles in aerobic energy producing pathways and in malate shuttle. In this study, the MDH-encoding gene for malate dehydrogenase mdh of B. abortus S2308 was cloned, sequenced and expressed. Western blot analysis demonstrated that MDH is an immunogenic membrane-associated protein. In addition, recombinant MDH showed sero-reactivity with 30 individual bovine B. abortus-positive sera by enzyme-linked immunosorbent assay, indicates that MDH may be used as a candidate marker for sero-diagnosis of brucellosis. Furthermore, MDH exhibits fibronectin and plasminogen-binding ability in immunoblotting assay. Inhibition assays on HeLa cells demonstrated that rabbit anti-serum against MDH significantly reduced both bacterial adherence and invasion abilities (p < 0.05), suggesting that MDH play a role in B. abortus colonization. Our results indicated that MDH is not only an immunogenic protein, but is also related to bacterial pathogenesis and may act as a new virulent factor, which will benefit for further understanding the MDH's roles in B. abortus metabolism, pathogenesis and immunity.

  14. 5-Lipoxygenase negatively regulates Th1 response during Brucella abortus infection in mice.

    Science.gov (United States)

    Fahel, Júlia Silveira; de Souza, Mariana Bueno; Gomes, Marco Túlio Ribeiro; Corsetti, Patricia P; Carvalho, Natalia B; Marinho, Fabio A V; de Almeida, Leonardo A; Caliari, Marcelo V; Machado, Fabiana Simão; Oliveira, Sergio Costa

    2015-03-01

    Brucella abortus is a Gram-negative bacterium that infects humans and cattle, causing a chronic inflammatory disease known as brucellosis. A Th1-mediated immune response plays a critical role in host control of this pathogen. Recent findings indicate contrasting roles for lipid mediators in host responses against infections. 5-Lipoxygenase (5-LO) is an enzyme required for the production of the lipid mediators leukotrienes and lipoxins. To determine the involvement of 5-LO in host responses to B. abortus infection, we intraperitoneally infected wild-type and 5-LO-deficient mice and evaluated the progression of infection and concomitant expression of immune mediators. Here, we demonstrate that B. abortus induced the upregulation of 5-LO mRNA in wild-type mice. Moreover, this pathogen upregulated the production of the lipid mediators leukotriene B4 and lipoxin A4 in a 5-LO-dependent manner. 5-LO-deficient mice displayed lower bacterial burdens in the spleen and liver and less severe liver pathology, demonstrating an enhanced resistance to infection. Host resistance paralleled an increased expression of the proinflammatory mediators interleukin-12 (IL-12), gamma interferon (IFN-γ), and inducible nitric oxide synthase (iNOS) during the course of infection. Moreover, we demonstrated that 5-LO downregulated the expression of IL-12 in macrophages during B. abortus infection. Our results suggest that 5-LO has a major involvement in B. abortus infection, by functioning as a negative regulator of the protective Th1 immune responses against this pathogen.

  15. The role of TREM-2 in internalization and intracellular survival of Brucella abortus in murine macrophages.

    Science.gov (United States)

    Wei, Pan; Lu, Qiang; Cui, Guimei; Guan, Zhenhong; Yang, Li; Sun, Changjiang; Sun, Wanchun; Peng, Qisheng

    2015-02-15

    Triggering receptor expressed on myeloid cells-2 (TREM-2) is a cell surface receptor primarily expressed on macrophages and dendritic cells. TREM-2 functions as a phagocytic receptor for bacteria as well as an inhibitor of Toll like receptors (TLR) induced inflammatory cytokines. However, the role of TREM-2 in Brucella intracellular growth remains unknown. To investigate whether TREM-2 is involved in Brucella intracellular survival, we chose bone marrow derived macrophages (BMDMs), in which TREM-2 is stably expressed, as cell model. Colony formation Units (CFUs) assay suggests that TREM-2 is involved in the internalization of Brucella abortus (B. abortus) by macrophages, while silencing of TREM-2 decreases intracellular survival of B. abortus. To further study the underlying mechanisms of TREM-2-mediated bacterial intracellular survival, we examined the activation of B. abortus-infected macrophages through determining the kinetics of activation of the three MAPKs, including ERK, JNK and p38, and measuring TNFα production in response to lipopolysaccharide (LPS) of Brucella (BrLPS) or B. abortus stimulation. Our data show that TREM-2 deficiency promotes activation of Brucella-infected macrophages. Moreover, our data also demonstrate that macrophage activation promotes killing of Brucella by enhancing nitric oxygen (NO), but not reactive oxygen species (ROS) production, macrophage apoptosis or cellular death. Taken together, these findings provide a novel interpretation of Brucella intracellular growth through inhibition of NO production produced by TREM-2-mediated activated macrophages.

  16. N-Formyl-Perosamine Surface Homopolysaccharides Hinder the Recognition of Brucella abortus by Mouse Neutrophils.

    Science.gov (United States)

    Mora-Cartín, Ricardo; Chacón-Díaz, Carlos; Gutiérrez-Jiménez, Cristina; Gurdián-Murillo, Stephany; Lomonte, Bruno; Chaves-Olarte, Esteban; Barquero-Calvo, Elías; Moreno, Edgardo

    2016-06-01

    Brucella abortus is an intracellular pathogen of monocytes, macrophages, dendritic cells, and placental trophoblasts. This bacterium causes a chronic disease in bovines and in humans. In these hosts, the bacterium also invades neutrophils; however, it fails to replicate and just resists the killing action of these leukocytes without inducing significant activation or neutrophilia. Moreover, B. abortus causes the premature cell death of human neutrophils. In the murine model, the bacterium is found within macrophages and dendritic cells at early times of infection but seldom in neutrophils. Based on this observation, we explored the interaction of mouse neutrophils with B. abortus In contrast to human, dog, and bovine neutrophils, naive mouse neutrophils fail to recognize smooth B. abortus bacteria at early stages of infection. Murine normal serum components do not opsonize smooth Brucella strains, and neutrophil phagocytosis is achieved only after the appearance of antibodies. Alternatively, mouse normal serum is capable of opsonizing rough Brucella mutants. Despite this, neutrophils still fail to kill Brucella, and the bacterium induces cell death of murine leukocytes. In addition, mouse serum does not opsonize Yersinia enterocolitica O:9, a bacterium displaying the same surface polysaccharide antigen as smooth B. abortus Therefore, the lack of murine serum opsonization and absence of murine neutrophil recognition are specific, and the molecules responsible for the Brucella camouflage are N-formyl-perosamine surface homopolysaccharides. Although the mouse is a valuable model for understanding the immunobiology of brucellosis, direct extrapolation from one animal system to another has to be undertaken with caution.

  17. Brucella abortus is Prevalent in Both Humans and Animals in Bangladesh.

    Science.gov (United States)

    Rahman, A K M A; Saegerman, C; Berkvens, D; Melzer, F; Neubauer, H; Fretin, D; Abatih, E; Dhand, N; Ward, M P

    2017-01-09

    To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test-positive human sera and all animal samples were screened by Brucella genus-specific real-time PCR (RT-PCR), and positive samples were then tested by IS711 RT-PCR to detect B. abortus and B. melitensis DNA. Only B. abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B. abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B. melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh.

  18. Epidemiology of brucellosis in domestic animals caused by Brucella melitensis, Brucella suis and Brucella abortus.

    Science.gov (United States)

    Díaz Aparicio, E

    2013-04-01

    Brucellosis is a disease that causes severe economic losses for livestock farms worldwide. Brucella melitensis, B. abortus and B. suis, which are transmitted between animals both vertically and horizontally, cause abortion and infertility in their primary natural hosts - goats and sheep (B. melitensis), cows (B. abortus) and sows (B. suis). Brucella spp. infect not only their preferred hosts but also other domestic and wild animal species, which in turn can act as reservoirs of the disease for other animal species and humans. Brucellosis is therefore considered to be a major zoonosis transmitted by direct contact with animals and/or their secretions, or by consuming milk and dairy products.

  19. Murine and bovine γδ T cells enhance innate immunity against Brucella abortus infections.

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    Jerod A Skyberg

    Full Text Available γδ T cells have been postulated to act as a first line of defense against infectious agents, particularly intracellular pathogens, representing an important link between the innate and adaptive immune responses. Human γδ T cells expand in the blood of brucellosis patients and are active against Brucella in vitro. However, the role of γδ T cells in vivo during experimental brucellosis has not been studied. Here we report TCRδ(-/- mice are more susceptible to B. abortus infection than C57BL/6 mice at one week post-infection as measured by splenic colonization and splenomegaly. An increase in TCRγδ cells was observed in the spleens of B. abortus-infected C57BL/6 mice, which peaked at two weeks post-infection and occurred concomitantly with diminished brucellae. γδ T cells were the major source of IL-17 following infection and also produced IFN-γ. Depletion of γδ T cells from C57BL/6, IL-17Rα(-/-, and GMCSF(-/- mice enhanced susceptibility to B. abortus infection although this susceptibility was unaltered in the mutant mice; however, when γδ T cells were depleted from IFN-γ(-/- mice, enhanced susceptibility was observed. Neutralization of γδ T cells in the absence of TNF-α did not further impair immunity. In the absence of TNF-α or γδ T cells, B. abortus-infected mice showed enhanced IFN-γ, suggesting that they augmented production to compensate for the loss of γδ T cells and/or TNF-α. While the protective role of γδ T cells was TNF-α-dependent, γδ T cells were not the major source of TNF-α and activation of γδ T cells following B. abortus infection was TNF-α-independent. Additionally, bovine TCRγδ cells were found to respond rapidly to B. abortus infection upon co-culture with autologous macrophages and could impair the intramacrophage replication of B. abortus via IFN-γ. Collectively, these results demonstrate γδ T cells are important for early protection to B. abortus infections.

  20. Immune responses of bison and efficacy after booster vaccination with Brucella abortus strain RB51.

    Science.gov (United States)

    Olsen, S C; McGill, J L; Sacco, R E; Hennager, S G

    2015-04-01

    Thirty-one bison heifers were randomly assigned to receive saline or a single vaccination with 10(10) CFU of Brucella abortus strain RB51. Some vaccinated bison were randomly selected for booster vaccination with RB51 at 11 months after the initial vaccination. Mean antibody responses to RB51 were greater (P abortus strain 2308. The incidences of abortion and infection were greater (P Brucella organisms in all tissues, except in retropharyngeal and supramammary lymph nodes. Our study suggests that RB51 booster vaccination is an effective vaccination strategy for enhancing herd immunity against brucellosis in bison.

  1. Simultaneous subcutaneous and conjunctival administration of the influenza viral vector based Brucella abortus vaccine to pregnant heifers provides better protection against B. abortus 544 infection than the commercial B. abortus S19 vaccine.

    Science.gov (United States)

    Tabynov, Kaissar; Orynbayev, Mukhit; Renukaradhya, Gourapura J; Sansyzbay, Abylai

    2016-09-30

    In this study, we explored possibility of increasing the protective efficacy of our novel influenza viral vector based B. abortus vaccine (Flu-BA) in pregnant heifers by adapting an innovative method of vaccine delivery. We administered the vaccine concurrently via the conjunctival and subcutaneous routes to pregnant heifers, and these routes were previously tested individually. The Flu-BA vaccination of pregnant heifers (n=9) against a challenge B. abortus 544 infection provided protection from abortion, infection of heifers and fetuses/calves by 88.8%, 100% and 100%, respectively (alpha=0.004-0.0007 vs. negative control; n=7). Our candidate vaccine using this delivery method provided slightly better protection than the commercial B. abortus S19 vaccine in pregnant heifers (n=8), which provided protection from abortion, infection of heifers and fetuses/calves by 87.5%, 75% and 87.5%, respectively. This improved method of the Flu-BA vaccine administration is highly recommended for the recovery of farms which has high prevalence of brucellosis.

  2. Vaccination of Elk (Cervus canadensis) with Brucella abortus Strain RB51 Overexpressing Superoxide Dismutase and Glycosyltransferase Genes Does Not Induce Adequate Protection against Experimental Brucella abortus Challenge

    OpenAIRE

    2016-01-01

    In recent years, elk (Cervus canadensis) have been implicated as the source of Brucella abortus infection for numerous cattle herds in the Greater Yellowstone Area. In the face of environmental and ecological changes on the landscape, the range of infected elk is expanding. Consequently, the development of effective disease management strategies for wild elk herds is of utmost importance, not only for the prevention of reintroduction of brucellosis to cattle, but also for the overall health...

  3. Herd-level prevalence and associated risk factors for Toxoplasma gondii, Neospora caninum, Chlamydia abortus and bovine viral diarrhoea virus in commercial dairy and beef cattle in eastern, northern and northeastern China.

    Science.gov (United States)

    Sun, Wu-Wen; Meng, Qing-Feng; Cong, Wei; Shan, Xiao-Feng; Wang, Chun-Feng; Qian, Ai-Dong

    2015-11-01

    Although the seroprevalence of Toxoplasma gondii, Neospora caninum, Chlamydia abortus and bovine viral diarrhea virus infection in cattle have been reported in some areas in China, most of them were conducted with small number of cattle samples and very limited districts and neglected the assessment of herd management factors associated with herd-level prevalence of these pathogen infections. Thus, from September 2013 to December 2014, a large-scale seroprevalence study was conducted to determine the animal-level and herd-level seroprevalence and identify herd-level risk factors associated with these pathogen infections in 4487 cattle from 134 herds in five provinces (Heilongjiang, Jilin, Liaoning, Shandong, Hebei) and Inner Mongolia Autonomous Region of China. At animal level, the true prevalence of antibodies against T. gondii, N. caninum, C. abortus and bovine viral diarrhoea virus (BVDV) was 10.48, 17.14, 11.92 and 50.10%, respectively. At herd level, the true prevalence of antibodies against T. gondii, N. caninum, C. abortus and BVDV was 27.16, 29.10, 37.31 and 40.30%, respectively. Multivariate analysis of these characteristics showed that source of water and presence of felids were significantly associated with T. gondii infection in the studied cattle herds. Source of water was significantly associated with N. caninum infection in the studied cattle herds. While herd size and management system were significantly associated with BVDV infection in the studied cattle herds, this is the first report of herd-level prevalence and associated risk factors of T. gondii, N. caninum, C. abortus and BVDV infection in cattle in China.

  4. Molecular Epidemiology of Brucella abortus in Northern Ireland-1991 to 2012.

    Directory of Open Access Journals (Sweden)

    Adrian Allen

    Full Text Available Brucellosis is the most common bacterial zoonoses worldwide. Bovine brucellosis caused by Brucella abortus has far reaching animal health and economic impacts at both the local and national levels. Alongside traditional veterinary epidemiology, the use of molecular typing has recently been applied to inform on bacterial population structure and identify epidemiologically-linked cases of infection. Multi-locus variable number tandem repeat VNTR analysis (MLVA was used to investigate the molecular epidemiology of a well-characterised Brucella abortus epidemic in Northern Ireland involving 387 herds between 1991 and 2012.MLVA identified 98 unique B. abortus genotypes from disclosing isolates in the 387 herds involved in the epidemic. Clustering algorithms revealed the relatedness of many of these genotypes. Combined with epidemiological information on chronology of infection and geographic location, these genotype data helped to identify 7 clonal complexes which underpinned the outbreak over the defined period. Hyper-variability of some VNTR loci both within herds and individual animals led to detection of multiple genotypes associated with single outbreaks. However with dense sampling, these genotypes could still be associated with specific clonal complexes thereby permitting inference of epidemiological links. MLVA- based epidemiological monitoring data were congruent with an independent classical veterinary epidemiology study carried out in the same territory.MLVA is a useful tool in ongoing disease surveillance of B. abortus outbreaks, especially when combined with accurate epidemiological information on disease tracings, geographical clustering of cases and chronology of infection.

  5. Immune responses of bison and efficacy after booster vaccination with Brucella abortus strain RB51

    Science.gov (United States)

    Thirty-one bison heifers were randomly assigned to saline (control; n=7) or single vaccination (n=24) with 1010 CFU of B. abortus strain RB51 (RB51). Some vaccinated bison were randomly selected for booster vaccination with 10**10 CFU of RB51 at 11 months after initial vaccination (n=16). When comp...

  6. Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR

    Institute of Scientific and Technical Information of China (English)

    Reza Mirnejad; Reza Hosseini Doust; Reza Kachuei; Seied Mojtaba Mortazavi; Mehdi Khoobdel; Ali Ahamadi

    2012-01-01

    Objective:To evaluate simultaneous detection and differentiates of Brucella abortus(B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method. Methods:This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes. Results:The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands;therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results. Conclusions:The results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.

  7. In Vitro Antibacterial Effects of Five Volatile Oil Extracts Against Intramacrophage Brucella Abortus 544

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    Ayman Al-Mariri

    2012-06-01

    Full Text Available Background: Brucella abortus is a gram-negative facultative intracellular bacterium that can cause a highly contagious disease in sheep, goats, cattle and one-humped camels. It is responsible for one of the most important zoonosis in human. The aim of this study was to evaluate the role of Mentha piperita, Origanum majorana, Citrus lemon, Cinnamomum verum and Myristica fragrans essential volatile oil extracts on human macrophages infected by B. abortus 544. Methods: Essential volatile oil extracts from M. piperita, O. majorana, C. lemon, C. verum and M. fragrans were extracted. Human macrophages were cultured at a density of 2×105 cells per well in sterile 96-well microtiter plates, and infected with B. abortus 544 at a ratio of 1:100 bacteria/cell. Then essential volatile oil extracts were added at a concentration of 1%. At specified times; cells were washed, lysed with 0.1% Triton, and plated on 2YT agar to determine the number of intracellular bacteria. Results: Cinnamomum verum volatile oil at a concentration of 1% had the highest antibacterial activity against B. abortus 544 inside human macrophages. Its inhibitory effect observed from 24 h and continued till 144 h after the infection. Moreover, C. verum (0.1% in combination with 1% concentration of M. piperita, O. majorana, C. lemon or M. fragrans volatile oil extracts produced a synergistic inhibitory effect against B. abortus 544. Conclusion: The results indicate that, among the five selected oil extracts, C. verum volatile oil applied either separately or in combination with other oil extracts had the most effective antimicrobial activity against Brucella.

  8. A LysR-family transcriptional regulator required for virulence in Brucella abortus is highly conserved among the α-proteobacteria.

    Science.gov (United States)

    Sheehan, Lauren M; Budnick, James A; Blanchard, Catlyn; Dunman, Paul M; Caswell, Clayton C

    2015-10-01

    Small RNAs are principal elements of bacterial gene regulation and physiology. Two small RNAs in Brucella abortus, AbcR1 and AbcR2, are required for wild-type virulence. Examination of the abcR loci revealed the presence of a gene encoding a LysR-type transcriptional regulator flanking abcR2 on chromosome 1. Deletion of this lysR gene (bab1_1517) resulted in the complete loss of abcR2 expression while no difference in abcR1 expression was observed. The B. abortus bab1_1517 mutant strain was significantly attenuated in macrophages and mice, and bab1_1517 was subsequently named vtlR for virulence-associated transcriptional LysR-family regulator. Microarray analysis revealed three additional genes encoding small hypothetical proteins also under the control of VtlR. Electrophoretic mobility shift assays demonstrated that VtlR binds directly to the promoter regions of abcR2 and the three hypothetical protein-encoding genes, and DNase I footprint analysis identified the specific nucleotide sequence in these promoters that VtlR binds to and drives gene expression. Strikingly, orthologs of VtlR are encoded in a wide range of host-associated α-proteobacteria, and it is likely that the VtlR genetic system represents a common regulatory circuit critical for host-bacterium interactions.

  9. Structural, functional and immunogenic insights on Cu,Zn Superoxide Dismutase pathogenic virulence factors from Neisseria meningitidis and Brucella abortus

    Science.gov (United States)

    Bacterial pathogens Neisseria meningitidis and Brucella abortus pose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acute N. meningitidis infections remains high despite antibiotics, and brucellosis presents alimentary and he...

  10. Molecular typing of isolates obtained from aborted foetuses in Brucella-free Holstein dairy cattle herd after immunisation with Brucella abortus RB51 vaccine in Egypt.

    Science.gov (United States)

    Wareth, Gamal; Melzer, Falk; Böttcher, Denny; El-Diasty, Mohamed; El-Beskawy, Mohamed; Rasheed, Nesma; Schmoock, Gernot; Roesler, Uwe; Sprague, Lisa D; Neubauer, Heinrich

    2016-12-01

    Bovine brucellosis is endemic in Egypt in spite of application of surveillance and control measures. An increase of abortions was reported in a Holstein dairy cattle herd with 600 animals in Damietta governorate in Egypt after immunisation with Brucella (B.) abortus RB51 vaccine. Twenty one (10.6%) of 197 vaccinated cows aborted after 3 months. All aborted cows had been tested seronegative for brucellosis in the past 3 years. B. abortus was isolated from four foetuses. Conventional biochemical and bacteriological identification and polymerase chain reaction (PCR) confirmed two B. abortus biovar (bv.) 1 smooth and two B. abortus rough strains. None of the B. abortus isolates were identified as RB51. Genotyping analysis by multiple locus of variable number tandem repeats analysis based on 16 markers (MLVA-16) revealed two different profiles with low genetic diversity. B. abortus bv1 was introduced in the herd and caused abortions.

  11. Pesquisa de anticorpos anti-Brucella abortus e anti-Brucella ovis em ovinos no município de Uberlândia, MG

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    S.R.S. Salaberry

    2011-08-01

    Full Text Available The first epidemiologic inquiry to Brucella abortus (B. abortus and Brucella ovis (B. ovis was carried out in sheep from Uberlândia county, MG. A total of 334 blood serum samples of sheep from both sexes and different ages and breeds were collected in 12 farms. An epidemiologic questionnaire was applied for each farm. Tests for B. abortus and B. ovis antibodies were Buffered Acidified Antigen and Complement Fixation, respectively. None of the sheep was reactive to B. abortus and B. ovis; however, the adoption of sanitary measures is important to avoid the introduction of infections caused by these bacteria.

  12. Genotyping of Brucella melitensis and Brucella abortus strains currently circulating in Xinjiang, China.

    Science.gov (United States)

    Sun, Ming-Jun; Di, Dong-Dong; Li, Yan; Zhang, Zhi-Cheng; Yan, Hao; Tian, Li-Li; Jing, Zhi-Gang; Li, Jin-Ping; Jiang, Hai; Fan, Wei-Xing

    2016-10-01

    Brucellosis is a well-known zoonotic disease that can cause severe economic and healthcare losses. Xinjiang, one of the biggest livestock husbandry sectors in China, has gone through increasing incidence of brucellosis in cattle and small ruminants recently. In this paper, 50 B. melitensis strains and 9 B. abortus strains collected from across Xinjiang area (from 2010 to 2015) were genotyped using multiple locus variable-number tandem-repeat (VNTR) analysis (MLVA) and multi-locus sequence typing (MLST). Based on 8 loci (MLVA-8), 50 B. melitensis strains were classified into three genotypes. Genotypes 42 (n=38, 76%) and 63 (n=11, 22%) were part of the East Mediterranean group, and one genotype with pattern of 1-5-3-13-2-4-3-2 represents a single-locus variant from genotype 63. MLVA-16 resolved 50 B. melitensis strains into 28 genotypes, of which 15 are unique to Xinjiang and 10 are in common with those in adjacent country Kazakhstan and neighboring provinces of China. Minimum Spanning Tree (MST) analysis implies that B. melitensis strains collected from across Kazakhstan, Xinjiang and China areas may share a common origin. Nine B. abortus strains were sorted into three genotypes by MLVA-8, genotypes 36 (n=7, 77.8%), 86 (n=1, 11.1%) and a new genotype with pattern of 4-5-3-13-2-2-3-1. Each B. abortus strain showed distinct MLVA-16 genotypes, suggesting that B. abortus species may possess more genetic diversity than B. melitensis. Using MLST, most B. melitensis strains (n=49) were identified as sequence type ST8, and most B. abortus strains (n=8) were recognized as ST2. Two new sequence types, ST37 and ST38, represented by single strain from B. melitensis and B. abortus species respectively, were also detected in this study. These results could facilitate the pathogen surveillance in the forthcoming eradication programs and serve as a guide in source tracking in case of new outbreaks occur.

  13. Brucella abortus: inmunidad, vacunas y estrategias de prevención basadas en ácidos nucleicos Brucella abortus: immunity, vaccines and prevention strategies based on nucleic acids

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    R Rivers

    2006-01-01

    Full Text Available Brucelosis, enfermedad causada por la bacteria intracelular facultativa Brucella abortus, es una zoonosis ampliamente distribuida a nivel mundial que afecta principalmente al ganado bovino, causando esterilidad en machos y abortos en hembras en gestación. La resistencia depende del desarrollo de una inmunidad mediada por células, con la participación de células T CD4+ de tipo Th1, que secreten interferón gama (-INF, citosina que estimula la actividad bactericida por macrófagos y la actividad citotóxica de linfocitos T CD8+, que son capaces de matar células infectadas con Brucella. Brucella posee como componentes antigénicos importantes el lipopolisacárido (LPS y las proteínas, entre las que se destaca por su demostrada capacidad inmune la superóxido dismutasa (SOD. La prevención de la diseminación de la brucelosis se fundamenta en el desarrollo de vacunas eficientes contra B. abortus, utilizándose cepas atenuadas de Brucella abortus como la cepa 19, cepa 45/20 y la cepa RB51; vacunas subcelulares en base a antígenos que forman parte de la estructura de la bacteria y vacunas basadas en moléculas de ácidos nucleicos, como las vacunas ADN y las vacunas ARN. En la presente revisión se pretende dar una visión actualizada sobre la brucelosis, su patogenia y cuadro clínico. Se hace un análisis de las características genéticas, antigénicas e inmunológicas de Brucella. Luego, una exposición de las vacunas actualmente en uso para su prevención y los estudios con vacunas subcelulares para finalizar con las nuevas tendencias en la generación de vacunas, como las vacunas ADN y ARN para Brucella.Brucellosis, a disease caused by the intracellular facultative bacteria Brucella abortus, is a widely distributed zoonosis throughout the world, affecting mainly cattle and causing sterility in males and abortion in pregnant females. Protection against B. abortus relies on the activation of cellular immunity involving lymphocytes T CD4

  14. Prime-booster vaccination of cattle with an influenza viral vector Brucella abortus vaccine induces a long-term protective immune response against Brucella abortus infection.

    Science.gov (United States)

    Tabynov, Kaissar; Yespembetov, Bolat; Ryskeldinova, Sholpan; Zinina, Nadezhda; Kydyrbayev, Zhailaubay; Kozhamkulov, Yerken; Inkarbekov, Dulat; Sansyzbay, Abylai

    2016-01-20

    This study analyzed the duration of the antigen-specific humoral and T-cell immune responses and protectiveness of a recently-developed influenza viral vector Brucella abortus (Flu-BA) vaccine expressing Brucella proteins Omp16 and L7/L12 and containing the adjuvant Montadine Gel01 in cattle. At 1 month post-booster vaccination (BV), both humoral (up to 3 months post-BV; GMT IgG ELISA titer 214±55 to 857±136, with a prevalence of IgG2a over IgG1 isotype antibodies) and T-cell immune responses were observed in vaccinated heifers (n=35) compared to control animals (n=35, injected with adjuvant/PBS only). A pronounced T-cell immune response was induced and maintained for 12 months post-BV, as indicated by the lymphocyte stimulation index (2.7±0.4 to 10.1±0.9 cpm) and production of IFN-γ (13.7±1.7 to 40.0±3.0 ng/ml) at 3, 6, 9, and 12 months post-BV. Prime-boost vaccination provided significant protection against B. abortus infection at 3, 6, 9 and 12 months (study duration) post-BV (7 heifers per time point; alpha=0.03-0.01 vs. control group). Between 57.1 and 71.4% of vaccinated animals showed no signs of B. abortus infection (or Brucella isolation) at 3, 6, 9 and 12 months post-BV; the severity of infection, as indicated by the index of infection (P=0.0003 to Brucella colonization (P=0.03 to abortus infection was also observed among pregnant vaccinated heifers (alpha=0.03), as well as their fetuses and calves (alpha=0.01), for 12 months post-BV. Additionally, 71.4% of vaccinated heifers calved successfully whereas all pregnant control animals aborted (alpha=0.01). Prime-boost vaccination of cattle with Flu-BA induces an antigen-specific humoral and pronounced T cell immune response and most importantly provides good protectiveness, even in pregnant heifers, for at least 12 months post-BV.

  15. Chlamydophila abortus infection in the mouse: a useful model of the ovine disease.

    Science.gov (United States)

    Caro, M R; Buendía, A J; Del Rio, L; Ortega, N; Gallego, M C; Cuello, F; Navarro, J A; Sanchez, J; Salinas, J

    2009-03-16

    Chlamydophila (C.) abortus is an obligate intracellular bacterium able to colonize the placenta of several species of mammals, which may induce abortion in the last third of pregnancy. The infection affects mainly small ruminants resulting in major economic losses in farming industries worldwide. Furthermore, its zoonotic risk has been reported in pregnant farmers or abattoir workers. Mouse models have been widely used to study both the pathology of the disease and the role of immune cells in controlling infection. Moreover, this animal experimental model has been considered a useful tool to evaluate new vaccine candidates and adjuvants that could prevent abortion and reduce fetal death. Future studies using these models will provide and reveal information about the precise mechanisms in the immune response against C. abortus and will increase the knowledge about poorly understood issues such as chlamydial persistence.

  16. A case of unusual septic knee arthritis with Brucella abortus after arthroscopic meniscus surgery.

    Science.gov (United States)

    Lee, Keun Hwa; Kang, Hyunseong; Kim, Taejung; Choi, Sungwook

    2016-01-01

    We present a 51-year-old male patient with Brucella abortus septic arthritis in the right knee following arthroscopic meniscus surgery. He had eaten a traditional dish of raw minced cattle conceptus (bovine fetus) that was prepared after the cow was slaughtered. Despite treatment with empirical antibiotics and debridement of the postoperative surgical wound, the infection persisted without improvement. Polymerase chain reaction sequencing identified Brucella abortus from tissue samples obtained from the patient. After confirmation of the diagnosis of brucellar infection, antibiotics were replaced with doxycycline and rifampin, which were used for 4 months. In patients with a non-specific arthralgia who eat raw meat or live close to animals, it is important to consider the possibility of septic arthritis due to infection with Brucella spp.

  17. Overview of the activity of a Brucella abortus preparation, Bru-Pel.

    Science.gov (United States)

    Youngner, J S; Feingold, D S; Keleti, G

    1978-11-01

    The properties of a nonviable, aqueous ether-extracted Brucela abortus preparation, Bru-Pel, are described. In addition to inducing a "virus-type" interferon response and protecting mice against challenge with otherwise lethal doses of Semliki Forest virus, Bru-Pel is demonstrated to have potent antitumor properties in mice. These antitumor effects appear to be mediated by an increase in nonspecific resistance similar to that seen with other experimental antitumor agents.

  18. Effects of gamma radiation and azathioprine on Brucella abortus infection in BALB/c mice

    Energy Technology Data Exchange (ETDEWEB)

    Elzer, P.H.; Rowe, G.E.; Enright, F.M.; Winter, A.J. (Department of Veterinary Microbiology, Immunology and Parasitology, College of Veterinary Medicine, Cornell University, Ithaca, NY (United States))

    1991-06-01

    Sublethal irradiation of BALB/c mice 4 hours prior to inoculation with 5 {times} 10(4) virulent Brucella abortus, caused significant (P less than 0.01) reductions in bacterial numbers in comparison with numbers in unirradiated controls. Numbers of brucellae in the spleen were significantly lower by 5 days after inoculation and decreased thereafter, so that at 2 and 3 weeks after inoculation, there were up to 1,000-fold fewer organisms in the spleen of irradiated mice. The number of brucellae in the spleen increased in irradiated mice thereafter. The course of events in the liver was similar, but developed more slowly, and peak differences in bacterial numbers were about 1 log less. These phenomena were not attributable to differences in implantation of brucellae in the liver or spleen, nor to an abnormal distribution of organisms in other organs of irradiated mice. Irradiation of mice during the plateau phase of infection also resulted in significant (P less than 0.05) reductions in bacterial counts in the spleen during the succeeding 4 weeks. Macrophage activation in the spleen, measured by a Listeria monocytogenes-killing assay, was significantly (P less than 0.01) increased by irradiation alone at 1 week after inoculation and at that time was significantly (P less than 0.01) greater in B abortus-infected, irradiated mice than in B abortus-infected controls. Histologic, cytologic, and immunologic studies revealed that the decrease in numbers of organisms between 1 and 2 weeks after inoculation in irradiated mice occurred at a time when their immune response to B abortus was suppressed and when numbers of neutrophils and monocytes infiltrating the spleen were significantly (P less than 0.01) diminished.

  19. Brucella abortus Induces the Premature Death of Human Neutrophils through the Action of Its Lipopolysaccharide.

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    Elías Barquero-Calvo

    2015-05-01

    Full Text Available Most bacterial infections induce the activation of polymorphonuclear neutrophils (PMNs, enhance their microbicidal function, and promote the survival of these leukocytes for protracted periods of time. Brucella abortus is a stealthy pathogen that evades innate immunity, barely activates PMNs, and resists the killing mechanisms of these phagocytes. Intriguing clinical signs observed during brucellosis are the low numbers of Brucella infected PMNs in the target organs and neutropenia in a proportion of the patients; features that deserve further attention. Here we demonstrate that B. abortus prematurely kills human PMNs in a dose-dependent and cell-specific manner. Death of PMNs is concomitant with the intracellular Brucella lipopolysaccharide (Br-LPS release within vacuoles. This molecule and its lipid A reproduce the premature cell death of PMNs, a phenomenon associated to the low production of proinflammatory cytokines. Blocking of CD14 but not TLR4 prevents the Br-LPS-induced cell death. The PMNs cell death departs from necrosis, NETosis and classical apoptosis. The mechanism of PMN cell death is linked to the activation of NADPH-oxidase and a modest but steadily increase of ROS mediators. These effectors generate DNA damage, recruitments of check point kinase 1, caspases 5 and to minor extent of caspase 4, RIP1 and Ca++ release. The production of IL-1β by PMNs was barely stimulated by B. abortus infection or Br-LPS treatment. Likewise, inhibition of caspase 1 did not hamper the Br-LPS induced PMN cell death, suggesting that the inflammasome pathway was not involved. Although activation of caspases 8 and 9 was observed, they did not seem to participate in the initial triggering mechanisms, since inhibition of these caspases scarcely blocked PMN cell death. These findings suggest a mechanism for neutropenia in chronic brucellosis and reveal a novel Brucella-host cross-talk through which B. abortus is able to hinder the innate function of PMN.

  20. Epidemiology of Brucellosis and Genetic Diversity of Brucella abortus in Kazakhstan

    Science.gov (United States)

    Shevtsova, Elena; Shevtsov, Alexandr; Mukanov, Kasim; Filipenko, Maxim; Kamalova, Dinara; Sytnik, Igor; Syzdykov, Marat; Kuznetsov, Andrey; Akhmetova, Assel; Zharova, Mira; Karibaev, Talgat; Tarlykov, Pavel; Ramanculov, Erlan

    2016-01-01

    Brucellosis is a major zoonotic infection in Kazakhstan. However, there is limited data on its incidence in humans and animals, and the genetic diversity of prevalent strains is virtually unstudied. Additionally, there is no detailed overview of Kazakhstan brucellosis control and eradication programs. Here, we analyzed brucellosis epidemiological data, and assessed the effectiveness of eradication strategies employed over the past 70 years to counteract this infection. We also conducted multiple loci variable-number tandem repeat analysis (MLVA) of Brucella abortus strains found in Kazakhstan. We analyzed official data on the incidence of animal brucellosis in Kazakhstan. The records span more than 70 years of anti-brucellosis campaigns, and contain a brief description of the applied control strategies, their effectiveness, and their impact on the incidence in humans. The MLVA-16 method was used to type 94 strains of B. abortus and serial passages of B. abortus 82, a strain used in vaccines. MLVA-8 and MLVA-11 analyses clustered strains into a total of four and seven genotypes, respectively; it is the first time that four of these genotypes have been described. MLVA-16 analysis divided strains into 28 distinct genotypes having genetic similarity coefficient that varies from 60 to100% and a Hunter & Gaston diversity index of 0.871. MST analysis reconstruction revealed clustering into "Kazakhstani-Chinese (Central Asian)", "European" and "American" lines. Detection of multiple genotypes in a single outbreak confirms that poorly controlled trade of livestock plays a crucial role in the spread of infection. Notably, the MLVA-16 profile of the B. abortus 82 strain was unique and did not change during 33 serial passages. MLVA genotyping may thus be useful for epidemiological monitoring of brucellosis, and for tracking the source(s) of infection. We suggest that countrywide application of MLVA genotyping would improve the control of brucellosis in Kazakhstan. PMID

  1. Brucella abortus Induces the Premature Death of Human Neutrophils through the Action of Its Lipopolysaccharide.

    Science.gov (United States)

    Barquero-Calvo, Elías; Mora-Cartín, Ricardo; Arce-Gorvel, Vilma; de Diego, Juana L; Chacón-Díaz, Carlos; Chaves-Olarte, Esteban; Guzmán-Verri, Caterina; Buret, Andre G; Gorvel, Jean-Pierre; Moreno, Edgardo

    2015-05-01

    Most bacterial infections induce the activation of polymorphonuclear neutrophils (PMNs), enhance their microbicidal function, and promote the survival of these leukocytes for protracted periods of time. Brucella abortus is a stealthy pathogen that evades innate immunity, barely activates PMNs, and resists the killing mechanisms of these phagocytes. Intriguing clinical signs observed during brucellosis are the low numbers of Brucella infected PMNs in the target organs and neutropenia in a proportion of the patients; features that deserve further attention. Here we demonstrate that B. abortus prematurely kills human PMNs in a dose-dependent and cell-specific manner. Death of PMNs is concomitant with the intracellular Brucella lipopolysaccharide (Br-LPS) release within vacuoles. This molecule and its lipid A reproduce the premature cell death of PMNs, a phenomenon associated to the low production of proinflammatory cytokines. Blocking of CD14 but not TLR4 prevents the Br-LPS-induced cell death. The PMNs cell death departs from necrosis, NETosis and classical apoptosis. The mechanism of PMN cell death is linked to the activation of NADPH-oxidase and a modest but steadily increase of ROS mediators. These effectors generate DNA damage, recruitments of check point kinase 1, caspases 5 and to minor extent of caspase 4, RIP1 and Ca++ release. The production of IL-1β by PMNs was barely stimulated by B. abortus infection or Br-LPS treatment. Likewise, inhibition of caspase 1 did not hamper the Br-LPS induced PMN cell death, suggesting that the inflammasome pathway was not involved. Although activation of caspases 8 and 9 was observed, they did not seem to participate in the initial triggering mechanisms, since inhibition of these caspases scarcely blocked PMN cell death. These findings suggest a mechanism for neutropenia in chronic brucellosis and reveal a novel Brucella-host cross-talk through which B. abortus is able to hinder the innate function of PMN.

  2. A rapid cycleave PCR method for distinguishing the vaccine strain Brucella abortus A19 in China.

    Science.gov (United States)

    Nan, Wenlong; Zhang, Yueyong; Tan, Pengfei; Xu, Zouliang; Chen, Yuqi; Mao, Kairong; Chen, Yiping

    2016-05-01

    Brucellosis is a widespread zoonotic disease caused by Brucella spp. Immunization with attenuated vaccines has proved to be an effective method of prevention; however, it may also interfere with diagnosis. Brucella abortus strain A19, which is homologous to B. abortus strain S19, is widely used for the prevention of bovine brucellosis in China. For effective monitoring of the control of brucellosis, it is essential to distinguish A19 from field strains. Single-nucleotide polymorphism-based assays offer a new approach to such discrimination studies. In the current study, we developed a cycleave PCR assay that successfully distinguished attenuated vaccine strains A19 and S19 from 22 strains of B. abortus and 57 strains of 5 other Brucella species. The assay gave a negative reaction with 4 non-Brucella species. The minimum sensitivity of the assay, evaluated using 10-fold dilutions of chromosomal DNA, was 7.6 fg for the A19 strain and 220 fg for the single non-A19/non-S19 Brucella strain tested (B. abortus 104M). The assay was also reproducible (intra- and interassay coefficients of variation: 0.003-0.01 and 0.004-0.025, respectively). The cycleave assay gave an A19/S19-specific reaction in 3 out of 125 field serum samples, with the same 3 samples being positive in an alternative A19/S19-specific molecular assay. The cycleave assay gave a total of 102 Brucella-specific reactions (3 being the A19/S19-specific reactions), whereas an alternative Brucella-specific assay gave 92 positive reactions (all also positive in the cycleave assay). Therefore, this assay represents a simple, rapid, sensitive, and specific tool for use in brucellosis control.

  3. Simultaneous differential detection of Chlamydophila abortus, Chlamydophila pecorum and Coxiella burnetii from aborted ruminant's clinical samples using multiplex PCR

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    Rodolakis Annie

    2009-07-01

    Full Text Available Abstract Background Chlamydiosis and Q fever, two zoonosis, are important causes of ruminants' abortion around the world. They are caused respectively by strictly intracellular and Gram negative bacterium Chlamydophila abortus (Cp. abortus and Coxiella burnetii (C. burnetii. Chlamydophila pecorum (Cp. pecorum is commonly isolated from the digestive tract of clinically inconspicuous ruminants but the abortive and zoonotic impact of this bacterium is still unknown because Cp. pecorum is rarely suspected in abortion cases of small ruminants. We have developed a multiplex PCR (m-PCR for rapid simultaneous differential detection of Cp. abortus, Cp. pecorum and C. burnetii in clinical samples taken from infected animals. Results Specific PCR primers were designed and a sensitive and specific m-PCR was developed to detect simultaneously, in one tube reaction, three specific fragments of 821, 526 and 687-bp long for Cp. abortus, Cp. pecorum and C. burnetii respectively. This m-PCR assay was performed on 253 clinical samples taken from infected ruminant's flocks that have showed problems of abortion diseases. Thus, 67 samples were infected by either one of the three pathogens: 16 (13 vaginal swabs and 3 placentas were positive for Cp. abortus, 2 were positive for Cp. pecorum (1 vaginal swab and 1 placenta and 49 samples (33 vaginal swabs, 11 raw milks, 4 faeces and 1 placenta were positive for C. burnetii. Two vaginal swabs were m-PCR positive of both Cp. abortus and C. burnetii and none of the tested samples was shown to be infected simultaneously with the three pathogens. Conclusion We have successfully developed a rapid multiplex PCR that can detect and differentiate Cp. abortus, Cp. pecorum and C. burnetii; with a good sensitivity and specificity. The diagnosis of chlamydiosis and Q fever may be greatly simplified and performed at low cost. In addition, the improvement in diagnostic techniques will enhance our knowledge regarding the prevalence and

  4. Purification and properties of Cu-Zn superoxide dismutase extracted from Brucella abortus strain 19

    Energy Technology Data Exchange (ETDEWEB)

    Tabatabai, L.B. (ARS-USDA, Ames, IA (United States))

    1991-03-11

    Recent work showed that a recombinant 20 kDa protein from Brucella abortus expressed in E. coli is a Cu-Zn superoxide dismutase (SOD). Western blot and ELISA results indicated that cattle with brucellosis have antibody to SOD. Here the authors report the purification and properties of the native B. abortus Cu-Zn SOD. SOD was extracted from methanol-killed Brucella abortus strain 19 with 0.1 M sodium citrate-1.0 M sodium chloride solution. The extract was dialyzed and protein precipitated by ammonium sulfate at 70-100% saturation was collected. The SOD was purified by HPLC anion exchange chromatography. SOD activity was assayed with a coupled enzyme assay using xanthine oxidase-cytochrome C reduction assay. The authors determined that the Brucella SOD is present in two molecular forms both inhibitable with KCN with Ki's of 0.32 mM and 4.98 mM, respectively. No other form of SOD was identified in the extract. Polyclonal antibody to SOD and polyclonal antibody to SOD synthetic peptide residues 134-143 inhibited SOD activity by 50% and 13%, respectively. Both SOD and the synthetic peptide inhibited binding of anti-SOD antibody to SOD by 60% and 20%, respectively. Based on these results the SOD and its amphipathic peptide will be considered as candidates for the design of synthetic multiple peptide vaccines and diagnostic reagents for bovine brucellosis.

  5. Brucella abortus Strain RB51 Vaccine: Immune Response after Calfhood Vaccination and Field Investigation in Italian Cattle Population

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    Manuela Tittarelli

    2008-01-01

    Full Text Available Immune response to Brucella abortus strain RB51 vaccine was measured in cattle vaccinated at calfhood. After an increase at day 6 post-vaccination (pv, the antibody level recorded in the 10 vaccinated animals remained constant for two months, and then progressively decreased. All vaccinated animals remained negative from day 162 pv to the end of the study (day 300 pv. Only at days 13 and 14 pv the RB51-CFT showed 100% sensitivity (credibility interval (CI 76.2%–100%. The results indicate that the possibility to use RB51-CFT for the identification of cattle vaccinated at calfhood with RB51 is limited in time. A field investigation was carried out on 26,975 sera collected on regional basis from the Italian cattle population. The study outcomes indicate that in case of RB51-CFT positive results observed in officially Brucellosis-free (OBF areas and, in any case, when an illegal use of RB51 vaccine is suspected, the use of the RB51-CFT alone is not sufficient to identify all the vaccinated animals. The design of a more sophisticated diagnostic protocol including an epidemiological investigation, the use of RB51-CFT, and the use of the skin test with RB51 as antigen is deemed more appropriate for the identification of RB51 vaccinated animals.

  6. Intranasal infection with Chlamydia abortus induces dose-dependent latency and abortion in sheep.

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    David Longbottom

    Full Text Available BACKGROUND: Latency is a key feature of the animal pathogen Chlamydia abortus, where infection remains inapparent in the non-pregnant animal and only becomes evident during a subsequent pregnancy. Often the first sign that an animal is infected is abortion occurring late in gestation. Despite this, little is understood of the underlying mechanisms that control latency or the recrudescence of infection that occurs during subsequent pregnancy. The aim of this study was to develop an experimental model of latency by mimicking the natural route of infection through the intranasal inoculation of non-pregnant sheep with C. abortus. METHODOLOGY/PRINCIPAL FINDINGS: Three groups of sheep (groups 1, 2 and 3 were experimentally infected with different doses of C. abortus (5×10(3, 5×10(5 and 5×10(7 inclusion forming units (IFU, respectively prior to mating and monitored over 2 breeding cycles for clinical, microbiological, pathological, immunological and serological outcomes. Two further groups received either negative control inoculum (group 4a,b or were inoculated subcutaneously on day 70 of gestation with 2×10(6 IFU C. abortus (group 5. Animals in groups 1, 2 and 5 experienced an abortion rate of 50-67%, while only one animal aborted in group 3 and none in group 4a,b. Pathological, microbiological, immunological and serological analyses support the view that the maternal protective immune response is influenced by initial exposure to the bacterium. CONCLUSIONS/SIGNIFICANCE: The results show that intranasal administration of non-pregnant sheep with a low/medium dose of C. abortus results in a latent infection that leads in a subsequent pregnancy to infection of the placenta and abortion. In contrast a high dose stimulates protective immunity, resulting in a much lower abortion rate. This model will be useful in understanding the mechanisms of infection underlying latency and onset of disease, as well as in the development of novel therapeutics and

  7. Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells

    Science.gov (United States)

    Bernardo Reyes, Alisha Wehdnesday; Simborio, Hannah Leah Tadeja; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, Won Gi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee

    2016-01-01

    Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis. PMID:26726017

  8. Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells.

    Science.gov (United States)

    Reyes, Alisha Wehdnesday Bernardo; Simborio, Hannah Leah Tadeja; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, Won Gi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee; Kim, Suk

    2016-09-30

    Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.

  9. Brucella abortus S19 and RB51 vaccine immunogenicity test: Evaluation of three mice (BALB/c, Swiss and CD-1) and two challenge strains (544 and 2308).

    Science.gov (United States)

    Miranda, Karina Leite; Dorneles, Elaine Maria Seles; Pauletti, Rebeca Barbosa; Poester, Fernando Padilla; Lage, Andrey Pereira

    2015-01-15

    The aim of the present study was to evaluate the use of different mouse strains (BALB/c, Swiss and CD-1) and different challenge strains (Brucella abortus 544 and 2308) in the study of B. abortus vaccine (S19 and RB51) immunogenicity test in the murine model. No significant difference in B. abortus vaccine potency assay was found with the use of B. abortus 544 or B. abortus 2308 as challenge strain. Results of variance analysis showed an interaction between treatment and mouse strain; therefore these parameters could not be compared separately. When CD-1 groups were compared, those vaccinated showed significantly lower counts than non-vaccinated ones (Pabortus strains 544 and 2308, can be used in immunogenicity tests of S19 and RB51 vaccines.

  10. Loop-Mediated Isothermal Amplification Assay Targeting the MOMP Gene for Rapid Detection of Chlamydia psittaci Abortus Strain

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    Guo-Zhen Lin, Fu-Ying Zheng, Ji-Zhang Zhou, Guang-Hua Wang, Xiao-An Cao, Xiao-Wei Gong and Chang-Qing Qiu*

    2012-05-01

    Full Text Available For rapid detection of the Chlamydia psittaci abortus strain, a loop-mediated isothermal amplification (LAMP assay was developed and evaluated in this study. The primers for the LAMP assay were designed on the basis of the main outer membrane protein (MOMP gene sequence of C. psittaci. Analysis showed that the assay could detect the abortus strain of C. psittaci with adequate specificity. The sensitivity of the test was the same as that of the nested-conventional PCR and higher than that of chick embryo isolation. Testing of 153 samples indicated that the LAMP assay could detect the genome of the C. psittaci abortus strain effectively in clinical samples. This assay is a useful tool for rapid diagnosis of C. psittaci infection in sheep, swine and cattle.

  11. Soroepidemiologia da brucelose canina causada por Brucella canis e Brucella abortus na cidade de Alfenas, MG Seroepidemiology of canine brucellosis caused by Brucella canis and Brucella abortus in Alfenas, MG, Brazil

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    A.C. Almeida

    2004-04-01

    Full Text Available The prevalence of canine brucellosis was evaluated in the city of Alfenas, MG through the technique of agarose gel imunodifusion for Brucella canis and slow serum agglutination test with 2-mercaptoetanol for Brucella abortus. The prevalence was of 14.2% and 2.8%, respectively, for B. canis and B. abortus. The positives, characterized by animals above one year of age (77.8%, and mongrel dogs (56.2%, showed a prevalence of 50 and 48% for males and females, respectively. The canine brucellosis was prevalent in the city principally in dogs of outskirts.

  12. Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts

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    Gamal Wareth

    2016-04-01

    Full Text Available Brucellosis is a debilitating zoonotic disease that affects humans and animals. The diagnosis of brucellosis is challenging, as accurate species level identification is not possible with any of the currently available serology-based diagnostic methods. The present study aimed at identifying Brucella (B. species-specific proteins from the closely related species B. abortus and B. melitensis using sera collected from naturally infected host species. Unlike earlier reported investigations with either laboratory-grown species or vaccine strains, in the present study, field strains were utilized for analysis. The label-free quantitative proteomic analysis of the naturally isolated strains of these two closely related species revealed 402 differentially expressed proteins, among which 63 and 103 proteins were found exclusively in the whole cell extracts of B. abortus and B. melitensis field strains, respectively. The sera from four different naturally infected host species, i.e., cattle, buffalo, sheep, and goat were applied to identify the immune-binding protein spots present in the whole protein extracts from the isolated B. abortus and B. melitensis field strains and resolved on two-dimensional gel electrophoresis. Comprehensive analysis revealed that 25 proteins of B. abortus and 20 proteins of B. melitensis were distinctly immunoreactive. Dihydrodipicolinate synthase, glyceraldehyde-3-phosphate dehydrogenase and lactate/malate dehydrogenase from B. abortus, amino acid ABC transporter substrate-binding protein from B. melitensis and fumarylacetoacetate hydrolase from both species were reactive with the sera of all the tested naturally infected host species. The identified proteins could be used for the design of serological assays capable of detecting pan-Brucella, B. abortus- and B. melitensis-specific antibodies.

  13. Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts.

    Science.gov (United States)

    Wareth, Gamal; Eravci, Murat; Weise, Christoph; Roesler, Uwe; Melzer, Falk; Sprague, Lisa D; Neubauer, Heinrich; Murugaiyan, Jayaseelan

    2016-04-30

    Brucellosis is a debilitating zoonotic disease that affects humans and animals. The diagnosis of brucellosis is challenging, as accurate species level identification is not possible with any of the currently available serology-based diagnostic methods. The present study aimed at identifying Brucella (B.) species-specific proteins from the closely related species B. abortus and B. melitensis using sera collected from naturally infected host species. Unlike earlier reported investigations with either laboratory-grown species or vaccine strains, in the present study, field strains were utilized for analysis. The label-free quantitative proteomic analysis of the naturally isolated strains of these two closely related species revealed 402 differentially expressed proteins, among which 63 and 103 proteins were found exclusively in the whole cell extracts of B. abortus and B. melitensis field strains, respectively. The sera from four different naturally infected host species, i.e., cattle, buffalo, sheep, and goat were applied to identify the immune-binding protein spots present in the whole protein extracts from the isolated B. abortus and B. melitensis field strains and resolved on two-dimensional gel electrophoresis. Comprehensive analysis revealed that 25 proteins of B. abortus and 20 proteins of B. melitensis were distinctly immunoreactive. Dihydrodipicolinate synthase, glyceraldehyde-3-phosphate dehydrogenase and lactate/malate dehydrogenase from B. abortus, amino acid ABC transporter substrate-binding protein from B. melitensis and fumarylacetoacetate hydrolase from both species were reactive with the sera of all the tested naturally infected host species. The identified proteins could be used for the design of serological assays capable of detecting pan-Brucella, B. abortus- and B. melitensis-specific antibodies.

  14. A T4SS Effector Targets Host Cell Alpha-Enolase Contributing to Brucella abortus Intracellular Lifestyle

    Science.gov (United States)

    Marchesini, María I.; Morrone Seijo, Susana M.; Guaimas, Francisco F.; Comerci, Diego J.

    2016-01-01

    Brucella abortus, the causative agent of bovine brucellosis, invades and replicates within cells inside a membrane-bound compartment known as the Brucella containing vacuole (BCV). After trafficking along the endocytic and secretory pathways, BCVs mature into endoplasmic reticulum-derived compartments permissive for bacterial replication. Brucella Type IV Secretion System (VirB) is a major virulence factor essential for the biogenesis of the replicative organelle. Upon infection, Brucella uses the VirB system to translocate effector proteins from the BCV into the host cell cytoplasm. Although the functions of many translocated proteins remain unknown, some of them have been demonstrated to modulate host cell signaling pathways to favor intracellular survival and replication. BPE123 (BAB2_0123) is a B. abortus VirB-translocated effector protein recently identified by our group whose function is yet unknown. In an attempt to identify host cell proteins interacting with BPE123, a pull-down assay was performed and human alpha-enolase (ENO-1) was identified by LC/MS-MS as a potential interaction partner of BPE123. These results were confirmed by immunoprecipitation assays. In bone-marrow derived macrophages infected with B. abortus, ENO-1 associates to BCVs in a BPE123-dependent manner, indicating that interaction with translocated BPE123 is also occurring during the intracellular phase of the bacterium. Furthermore, ENO-1 depletion by siRNA impaired B. abortus intracellular replication in HeLa cells, confirming a role for α-enolase during the infection process. Indeed, ENO-1 activity levels were enhanced upon B. abortus infection of THP-1 macrophagic cells, and this activation is highly dependent on BPE123. Taken together, these results suggest that interaction between BPE123 and host cell ENO-1 contributes to the intracellular lifestyle of B. abortus. PMID:27900285

  15. Identificación de la cepa vacunal Brucella abortus S19 en muestras de leche de vaca

    OpenAIRE

    Luary Carolina Martínez Chavarría; Antonio Verdugo Rodríguez; Rigoberto Hernández Castro

    2006-01-01

    La brucelosis es una enfermedad infectocontagiosa de origen bacteriano que afecta tanto al humano como a diferentes especies animales domésticas y silvestres. En la década pasada, la vacuna que se usaba ampliamente en los bovinos era la cepa vacunal B. abortus S19, que tiene una deleción en dos de los genes del operón ery que participa en el catabolismo del eritritol. En México, desde 1997 se aprobó la cepa B. abortus RB51 como vacuna para el ganado. Esta cepa presenta una secuencia de inserc...

  16. Molecular structure of the Brucella abortus metalloprotein RicA, a Rab2-binding virulence effector.

    Science.gov (United States)

    Herrou, Julien; Crosson, Sean

    2013-12-17

    The Gram-negative intracellular pathogen Brucella abortus is the causative agent of brucellosis, which is among the most common zoonoses globally. The B. abortus RicA protein binds the host-expressed guanosine nucleotide-binding protein, Rab2, and modulates B. abortus infection biology. We have solved the first X-ray crystal structure of RicA to 2.7 Å resolution and have quantified the affinity of RicA binding to human Rab2 in its GDP-bound and nucleotide-free forms. RicA adopts a classic γ-carbonic anhydrase (γ-CA) fold containing a left-handed β-helix followed by a C-terminal α-helix. Two homotrimers of RicA occupy the crystallographic asymmetric unit. Though no zinc was included in the purification or crystallization buffers, zinc is contained within the RicA crystals, as demonstrated by X-ray fluorescence spectroscopy. Electron density for a Zn(2+) ion coordinated by three histidine residues is evident in the putative active site of RicA. However, purified RicA preparations do not exhibit carbonic anhydrase activity, suggesting that Zn(2+) may not be the physiologically relevant metal cofactor or that RicA is not a bona fide carbonic anhydrase enzyme. Isothermal titration calorimetry (ITC) measurements of purified RicA binding to purified human Rab2 and GDP-Rab2 revealed similar equilibrium affinities (Kd ≈ 35 and 40 μM, respectively). This study thus defines RicA as a Zn(2+)-binding γ-carbonic anhydrase-like protein that binds the human membrane fusion/trafficking protein Rab2 with low micromolar affinity in vitro. These results support a model in which γ-CA family proteins may evolve unique cellular functions while retaining many of the structural hallmarks of archetypal γ-CA enzymes.

  17. Different resistance patterns of reference and field strains of Brucella abortus

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    Karina L. Miranda

    2015-03-01

    Full Text Available The aim of this study was to evaluate the growth of the B. abortus reference strains and field isolates on media containing different inhibitor agents. Reference strains were seeded on tryptose agar containing: i-erythritol (1.0 mg/mL, fuchsin (20 μg/mL and 80 μg/mL, thionin (2.5 μg/mL and 10 μg/mL, rifampicin (200 μg/mL and safranin O (200 μg/mL. Field isolates were tested only on media containing i-erythritol, rifampicin and thionin. Furthermore, each suspension was also inoculated on tryptose agar incubated in air, to test its ability to grow without CO2. Sensitivity to fuchsin was similar among reference strains evaluated. Growth of S19, 544 and 2308 but not RB51 were inhibited on media containing rifampicin. Medium with safranin O showed no inhibition for RB51, 544 and 2308, but it partially inhibited the S19 growth as well as medium containing i-erythritol. Treatment/control growth ratio for 2308 on tryptose agar containing thionin (2.5 μg/mL was approximatelly 1.0, whereas S19 and RB51 showed 0.85 and 0.89 ratios, respectively. Growth of 544, S19 and RB51 but not 2308 was completely inhibited on medium with thionin (10 μg/mL. All field strains grew on medium containing i-erythritol, but were completelly inhibited by rifampicin. With exception of A1 (B. abortus biovar 3 all field isolates grew on medium with thionin, although some strains showed a treatment/control growth ratio of 0.75–0.80 (10 μg/mL. These results showed that tryptose agar with thionin, i-erythritol or rifampicin could be useful for differentiating vaccine, challenge and field strains of B. abortus.

  18. Different resistance patterns of reference and field strains of Brucella abortus.

    Science.gov (United States)

    Miranda, Karina L; Dorneles, Elaine M S; Poester, Fernando P; Martins Filho, Paulo S; Pauletti, Rebeca B; Lage, Andrey P

    2015-03-01

    The aim of this study was to evaluate the growth of the B. abortus reference strains and field isolates on media containing different inhibitor agents. Reference strains were seeded on tryptose agar containing: i-erythritol (1.0 mg/mL), fuchsin (20 μg/mL and 80 μg/mL), thionin (2.5 μg/mL and 10 μg/mL), rifampicin (200 μg/mL) and safranin O (200 μg/mL). Field isolates were tested only on media containing i-erythritol, rifampicin and thionin. Furthermore, each suspension was also inoculated on tryptose agar incubated in air, to test its ability to grow without CO 2 . Sensitivity to fuchsin was similar among reference strains evaluated. Growth of S19, 544 and 2308 but not RB51 were inhibited on media containing rifampicin. Medium with safranin O showed no inhibition for RB51, 544 and 2308, but it partially inhibited the S19 growth as well as medium containing i-erythritol. Treatment/control growth ratio for 2308 on tryptose agar containing thionin (2.5 μg/mL) was approximatelly 1.0, whereas S19 and RB51 showed 0.85 and 0.89 ratios, respectively. Growth of 544, S19 and RB51 but not 2308 was completely inhibited on medium with thionin (10 μg/mL). All field strains grew on medium containing i-erythritol, but were completelly inhibited by rifampicin. With exception of A1 ( B. abortus biovar 3) all field isolates grew on medium with thionin, although some strains showed a treatment/control growth ratio of 0.75-0.80 (10 μg/mL). These results showed that tryptose agar with thionin, i-erythritol or rifampicin could be useful for differentiating vaccine, challenge and field strains of B. abortus.

  19. Proinflammatory Response of Human Trophoblastic Cells to Brucella abortus Infection and upon Interactions with Infected Phagocytes.

    Science.gov (United States)

    Fernández, Andrea G; Ferrero, Mariana C; Hielpos, M Soledad; Fossati, Carlos A; Baldi, Pablo C

    2016-02-01

    Trophoblasts are targets of infection by Brucella spp. but their role in the pathophysiology of pregnancy complications of brucellosis is unknown. Here we show that Brucella abortus invades and replicates in the human trophoblastic cell line Swan-71 and that the intracellular survival of the bacterium depends on a functional virB operon. The infection elicited significant increments of interleukin 8 (IL8), monocyte chemotactic protein 1 (MCP-1), and IL6 secretion, but levels of IL1beta and tumor necrosis factor-alpha (TNF-alpha) did not vary significantly. Such proinflammatory response was not modified by the absence of the Brucella TIR domain-containing proteins BtpA and BtpB. The stimulation of Swan-71 cells with conditioned medium (CM) from B. abortus-infected human monocytes (THP-1 cells) or macrophages induced a significant increase of IL8, MCP-1 and IL6 as compared to stimulation with CM from non-infected cells. Similar results were obtained when stimulation was performed with CM from infected neutrophils. Neutralization studies showed that IL1beta and/or TNF-alpha mediated the stimulating effects of CM from infected phagocytes. Reciprocally, stimulation of monocytes and neutrophils with CM from Brucella-infected trophoblasts increased IL8 and/or IL6 secretion. These results suggest that human trophoblasts may provide a local inflammatory environment during B. abortus infections either through a direct response to the pathogen or through interactions with monocytes/macrophages or neutrophils, potentially contributing to the pregnancy complications of brucellosis.

  20. Outbreak of laboratory-acquired Brucella abortus in Brazil: a case report

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    Ana Luisa Calixto Rodrigues

    2013-12-01

    Full Text Available Human brucellosis is an occupational disease affecting workers in slaughterhouses, butcher shops and the milk and dairy product industry as well as individuals who work in clinical or research laboratories. We report the first outbreak of a Brucella abortus infection in a Brazilian laboratory and compare the data obtained with reports available in the literature. Exposure was a result of damage to a biological safety cabinet and failure of the unidirectional airflow ventilation system. An epidemiological investigation identified 3 seroconverted individuals, 1 of whom had clinical manifestations and laboratory results compatible with infection at the time of exposure (n=11; attack rate=9.1%.

  1. TLR7 and TLR3 Sense Brucella abortus RNA to Induce Proinflammatory Cytokine Production but They Are Dispensable for Host Control of Infection

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    Campos, Priscila C.; Gomes, Marco Túlio R.; Guimarães, Erika S.; Guimarães, Gabriela; Oliveira, Sergio C.

    2017-01-01

    Brucella abortus is a Gram-negative, facultative intracellular bacterium that causes brucellosis, a worldwide zoonotic disease leading to undulant fever in humans and abortion in cattle. The immune response against this bacterium relies on the recognition of microbial pathogen-associated molecular patterns, such as lipoproteins, lipopolysaccharides, and DNA; however, the immunostimulatory potential of B. abortus RNA remains to be elucidated. Here, we show that dendritic cells (DCs) produce significant amounts of IL-12, IL-6, and IP-10/CXCL10, when stimulated with purified B. abortus RNA. IL-12 secretion by DCs stimulated with RNA depends on TLR7 while IL-6 depends on TLR7 and partially on TLR3. Further, only TLR7 plays a role in IL-12 production induced by B. abortus infection. Moreover, cytokine production in DCs infected with B. abortus or stimulated with bacterial RNA was reduced upon pretreatment with MAPK/NF-κB inhibitors. By confocal microscopy, we demonstrated that TLR7 is colocalized with B. abortus in LAMP-1+ Brucella-containing vacuoles. Additionally, type I IFN expression and IP-10/CXCL10 secretion in DCs stimulated with bacterial RNA were dependent on TLR3 and TLR7. Our results suggest that TLR3 and TLR7 are not required to control Brucella infection in vivo, but they play an important role on sensing B. abortus RNA in vitro. PMID:28167945

  2. The Effector Protein BPE005 from Brucella abortus Induces Collagen Deposition and Matrix Metalloproteinase 9 Downmodulation via Transforming Growth Factor β1 in Hepatic Stellate Cells.

    Science.gov (United States)

    Arriola Benitez, Paula Constanza; Rey Serantes, Diego; Herrmann, Claudia Karina; Pesce Viglietti, Ayelén Ivana; Vanzulli, Silvia; Giambartolomei, Guillermo Hernán; Comerci, Diego José; Delpino, María Victoria

    2015-12-14

    The liver is frequently affected in patients with active brucellosis. In the present study, we identified a virulence factor involved in the modulation of hepatic stellate cell function and consequent fibrosis during Brucella abortus infection. This study assessed the role of BPE005 protein from B. abortus in the fibrotic phenotype induced on hepatic stellate cells during B. abortus infection in vitro and in vivo. We demonstrated that the fibrotic phenotype induced by B. abortus on hepatic stellate (LX-2) cells was dependent on BPE005, a protein associated with the type IV secretion system (T4SS) VirB from B. abortus. Our results indicated that B. abortus inhibits matrix metalloproteinase 9 (MMP-9) secretion through the activity of the BPE005-secreted protein and induces concomitant collagen deposition by LX-2 cells. BPE005 is a small protein containing a cyclic nucleotide monophosphate binding domain (cNMP) that modulates the LX-2 cell phenotype through a mechanism that is dependent on the cyclic AMP (cAMP)/protein kinase A (PKA) signaling pathway. Altogether, these results indicate that B. abortus tilts LX-2 cells to a profibrogenic phenotype employing a functional T4SS and the secreted BPE005 protein through a mechanism that involves the cAMP and PKA signaling pathway.

  3. Investigation of the possible role of Chlamydophila abortus in reproductive failures in nrazilian herds of domestic ruminants

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    Francielle Gibson da Silva-Zacarias

    2009-11-01

    Full Text Available Chlamydophila abortus (C. abortus infection is related to reproductive failure in domestic ruminants. Although it has not been well characterized worldwide, this pathogen has already been identified in some European countries and in the USA. In Brazil, preliminary studies have shown serological evidence of C. abortus infection in herds with low antibody prevalence. Until now, the identification of C. abortus in biological samples from females presenting reproductive failures has not been described in Brazilian herds of domestic ruminants. The aim of this study was to evaluate the presence of the C. abortus in a collection of abortions from cattle (n=85, sheep (n=12, and goats (n=8, in samples of vaginal mucus from cows (n=13, sheep (n=90, and goats (n­=20, and in semen from sheep (n=10 and goats (n=5. The specimens (n=243 were evaluated using a PCR assay developed to amplify the 16S-23S rRNA intergenic space of C. abortus. A PCR assay with an internal control, which amplifies a fragment from the ND5 gene of bovine mitochondrial DNA, was used in order to evaluate the efficiency of the DNA extraction and of the PCR reaction. All biological samples (n=243 included in this study were negative for C. abortus in the PCR assay. The internal control enabled the amplification of a product from the bovine mitochondrial ND5 gene in all cattle abortion samples (n=85. Given the serological evidence indicating the presence of C. abortus infection in Brazilian herds of domestic ruminants, and considering the wide sampling evaluated, the failure to identify C. abortus in this survey suggests that the frequency of clinical signs in infected animals may be low or even absent.A infecção pela Chlamydophila abortus (C. abortus em ruminantes domésticos está relacionada com distúrbios reprodutivos. Apesar de ainda pouco estudada em todo o mundo, a infecção já foi identificada em alguns países europeus e também nos EUA. No Brasil, estudos preliminares

  4. Immunogenicity and protective efficacy of Brucella abortus recombinant protein cocktail (rOmp19+rP39) against B. abortus 544 and B. melitensis 16M infection in murine model.

    Science.gov (United States)

    Tadepalli, Ganesh; Singh, Amit Kumar; Balakrishna, Konduru; Murali, Harishchandra Sripathy; Batra, Harsh Vardhan

    2016-03-01

    In this study, the immunogenicity and protective efficacy of recombinant proteins Omp19 (rO) and P39 (rP) from Brucella abortus were evaluated individually and compared with the cocktail protein (rO+rP) against B. abortus 544 and Brucella melitensis 16M infection in BALB/c mouse model. Intra-peritoneal (I.P.) immunization with rO+rP cocktail developed substantially higher antibody titers predominant with Th1 mediated isotypes (IgG2a/2b). Western blot analysis using anti-rO+rP antibodies showed specific reactivity with native Omp19 (19 kDa) and P39 (39 kDa) among whole cell proteins of B. abortus and B. melitensis. Splenocytes extracted from rO+rP immunized mice induced significantly (Pabortus 544 (72.27%) and B. melitensis 16M (68.57%). On the other hand, individual anti-rO and anti-rP polysera resulted in relatively lesser protection against the pathogens (64.79%, 54.45% and 47.13%, 45.11%, respectively). Immunized group of mice when I.P. challenged with 5 × 10(4) CFU of B. abortus 544 and B. melitensis 16M were found significantly (PBrucella vaccine.

  5. Post-Abortion Syndrome and nursing approachesPost-Abortus Sendromu ve hemşirelik yaklaşımları

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    Yılda Arzu Aba

    2016-02-01

    Full Text Available The relation between the life outputs, are associated especially with the mental health outputs at Abortus and the period that follows it. Post-Abortus Syndrome (PAS is the concept of abortus trauma management that show up with the chronic or delayed symptoms as a result of the hidden emotional reactions that are experienced because of the post-abortus physical and psychological trauma. The individual, the systemic and the social factors that are existing at post-Abortus women affect PAS development. The interventions, in general, that are oriented towards mental problems of the individual at the PAS period, focused on the  individual’s sufficiency and efficiency for coping with all the problems that are faced in her life. There is no any program that is developed for post-Abortus women or their family in our country. It is important for the individual who is diagnosed with PAS to receive consultancy from a health professional for her recovery from the period.   Özet Abortus ve bunu izleyen süreçte yaşam sonuçları arasındaki ilişki özellikle mental sağlık sonuçlarıyla ilişkilendirilmektedir. Post-Abortus Sendromu (PAS, abortus sonrası fiziksel ve psikolojik travma nedeniyle deneyimlenen gizlenmiş duygusal tepkiler sonucu, kronik ya da gecikmiş belirtilerle ortaya çıkan abortus travmasını yönetebilme kavramıdır. Abortus sonrası kadınlarda var olan bireysel, sistemik ve sosyal faktörler PAS gelişimini etkilemektedir. PAS dönemindeki bireyin ruhsal sorunlarına yönelik yapılan müdahaleler genel olarak, bireyin yaşamında karşılaştığı tüm sorunlarla başetmesinde yeterli ve etkili hale gelmesine odaklanır. Ülkemizde abortus sonrası kadınlara ya da ailelerine yönelik geliştirilen herhangi bir program bulunmamaktadır. PAS tanısı alan bireyin süreci atlatabilmesi için sağlık profesyoneli tarafından danışmanlık alması önemlidir.

  6. A Live Vaccine from Brucella abortus Strain 82 for Control of Cattle Brucellosis in the Russian Federation

    Science.gov (United States)

    During the first half of the 20th century, widespread regulatory efforts to control cattle brucellosis (Brucella abortus) in the Union of Soviet Socialist Republics were essentially nonexistent, and control was limited to selective test and slaughter of serologic agglutination reactors. By the 1950...

  7. Efficacy of dart or booster vaccination with strain RB51 in protecting bison against experimental Brucella abortus challenge

    Science.gov (United States)

    Vaccination is an effective tool for reducing the prevalence of brucellosis in natural hosts. In this study, we characterized the efficacy of the Brucella abortus strain RB51 (RB51) vaccine in bison when delivered by single intramuscular vaccination (Hand RB51), single pneumatic dart delivery (Dart ...

  8. Draft Genome Sequence of the Intermediate Rough Vaccine Strain Brucella abortus S19Δper Mutant

    Science.gov (United States)

    Chaudhuri, Pallab; Goswami T, Tapas K.; Lalsiamthara, Jonathan; Kaur, Gurpreet; Vishnu, Udayakumar S.; Sankarasubramanian, Jagadesan; Gunasekaran, Paramasamy

    2015-01-01

    Here, we report the genome sequence of the intermediate rough vaccine strain mutant, Brucella abortus S19Δper. The length of the draft genome was 3,271,238 bp, with 57.2% G+C content. A total of 3,204 protein-coding genes and 56 RNA genes were predicted. PMID:26564050

  9. Host-pathogen interactions in specific pathogen-free chickens following aerogenous infection with Chlamydia psittaci and Chlamydia abortus.

    Science.gov (United States)

    Kalmar, Isabelle; Berndt, Angela; Yin, Lizi; Chiers, Koen; Sachse, Konrad; Vanrompay, Daisy

    2015-03-15

    Although Chlamydia (C.) psittaci infections are recognized as an important factor causing economic losses and impairing animal welfare in poultry production, the specific mechanisms leading to severe clinical outcomes are poorly understood. In the present study, we comparatively investigated pathology and host immune response, as well as systemic dissemination and expression of essential chlamydial genes in the course of experimental aerogeneous infection with C. psittaci and the closely related C. abortus, respectively, in specific pathogen-free chicks. Clinical signs appeared sooner and were more severe in the C. psittaci-infected group. Compared to C. abortus infection, more intense systemic dissemination of C. psittaci correlated with higher and faster infiltration of immune cells, as well as more macroscopic lesions and epithelial pathology, such as hyperplasia and erosion. In thoracic air sac tissue, mRNA expression of immunologically relevant factors, such as IFN-γ, IL-1β, IL-6, IL-17, IL-22, LITAF and iNOS was significantly stronger up-regulated in C. psittaci- than in C. abortus-infected birds between 3 and 14 days post-infection. Likewise, transcription rates of the chlamydial genes groEL, cpaf and ftsW were consistently higher in C. psittaci during the acute phase. These findings illustrate that the stronger replication of C. psittaci in its natural host also evoked a more intense immune response than in the case of C. abortus infection.

  10. Dextran sulfate sodium upregulates MAPK signaling for the uptake and subsequent intracellular survival of Brucella abortus in murine macrophages.

    Science.gov (United States)

    Reyes, Alisha Wehdnesday Bernardo; Arayan, Lauren Togonon; Simborio, Hannah Leah Tadeja; Hop, Huynh Tan; Min, WonGi; Lee, Hu Jang; Kim, Dong Hee; Chang, Hong Hee; Kim, Suk

    2016-02-01

    Brucellosis is one of the major zoonoses worldwide that inflicts important health problems in animal and human. Here, we demonstrated that dextran sulfate sodium (DSS) significantly increased adhesion of Brucella (B.) abortus in murine macrophages compared to untreated cells. Even without infection, Brucella uptake into macrophages increased and F-actin reorganization was induced compared with untreated cells. Furthermore, DSS increased the phosphorylation of MAPKs (ERK1/2 and p38α) in Brucella-infected, DSS-treated cells compared with the control cells. Lastly, DSS markedly increased the intracellular survival of Brucella abortus in macrophages by up to 48 h. These results suggest that DSS enhanced the adhesion and phagocytosis of B. abortus into murine macrophages by stimulating the MAPK signaling proteins phospho-ERK1/2 and p38α and that DSS increased the intracellular survival of B. abortus by inhibiting colocalization of Brucella-containing vacuoles (BCVs) with the late endosome marker LAMP-1. This study emphasizes the enhancement of the phagocytic and intracellular modulatory effects of DSS, which may suppress the innate immune system and contribute to prolonged Brucella survival and chronic infection.

  11. Development of a dual vaccine for prevention of Brucella abortus infection and Escherichia coli O157:H7 intestinal colonization.

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    Iannino, Florencia; Herrmann, Claudia K; Roset, Mara S; Briones, Gabriel

    2015-05-05

    Zoonoses that affect human and animal health have an important economic impact. In the study now presented, a bivalent vaccine has been developed that has the potential for preventing the transmission from cattle to humans of two bacterial pathogens: Brucella abortus and Shiga toxin-producing Escherichia coli (STEC). A 66kDa chimeric antigen, composed by EspA, Intimin, Tir, and H7 flagellin (EITH7) from STEC, was constructed and expressed in B. abortus Δpgm vaccine strain (BabΔpgm). Mice orally immunized with BabΔpgm(EITH7) elicited an immune response with the induction of anti-EITH7 antibodies (IgA) that clears an intestinal infection of E. coli O157:H7 three times faster (t=4 days) than mice immunized with BabΔpgm carrier strain (t=12 days). As expected, mice immunized with BabΔpgm(EITH7) strain also elicited a protective immune response against B. abortus infection. A Brucella-based vaccine platform is described capable of eliciting a combined protective immune response against two bacterial pathogens with diverse lifestyles-the intracellular pathogen B. abortus and the intestinal extracellular pathogen STEC.

  12. Comparison of abortion and infection after experimental challenge of pregnant bison and cattle with Brucella abortus strain 2308

    Science.gov (United States)

    A comparative study was conducted using data from naive bison (n=45) and cattle (n=46) from 8 and 6 studies, respectively, in which a standardized Brucella abortus strain 2308 experimental challenge was administered. The incidence of abortion, fetal infection, uterine or mammary infection, or infec...

  13. Brucella abortus mutants lacking ATP-binding cassette transporter proteins are highly attenuated in virulence and confer protective immunity against virulent B. abortus challenge in BALB/c mice.

    Science.gov (United States)

    Truong, Quang Lam; Cho, Youngjae; Park, Soyeon; Park, Bo-Kyoung; Hahn, Tae-Wook

    2016-06-01

    Brucella abortus RB51 is an attenuated vaccine strain that has been most frequently used for bovine brucellosis. Although it is known to provide good protection in cattle, it still has some drawbacks including resistance to rifampicin, residual virulence and pathogenicity in humans. Thus, there has been a continuous interest on new safe and effective bovine vaccine candidates. In the present study, we have constructed unmarked mutants by deleting singly cydD and cydC genes, which encode ATP-binding cassette transporter proteins, from the chromosome of the virulent Brucella abortus isolate from Korean cow (referred to as IVK15). Both IVK15ΔcydD and ΔcydC mutants showed increased sensitivity to metal ions, hydrogen peroxide and acidic pH, which are mimic to intracellular environment during host infection. Additionally, the mutants exhibited a significant growth defect in RAW264.7 cells and greatly attenuated in mice. Vaccination of mice with either IVK15ΔcydC or IVK15ΔcydD mutant could elicit an anti-Brucella specific immunoglobulin G (IgG) and IgG subclass responses as well as enhance the secretion of interferon-gamma, and provided better protection against challenge with B. abortus strain 2308 than with the commercial B. abortus strain RB51 vaccine. Collectively, these results suggest that both IVK15ΔcydC and IVK15ΔcydD mutants could be an attenuated vaccine candidate against B. abortus.

  14. Enzymatic Activity Analysis and Catalytic Essential Residues Identification of Brucella abortus Malate Dehydrogenase

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    Xiangan Han

    2014-01-01

    Full Text Available Malate dehydrogenase (MDH plays important metabolic roles in bacteria. In this study, the recombinant MDH protein (His-MDH of Brucella abortus was purified and its ability to catalyze the conversion of oxaloacetate (OAA to L-malate (hereon referred to as MDH activity was analyzed. Michaelis Constant (Km and Maximum Reaction Velocity (Vmax of the reaction were determined to be 6.45×10−3 M and 0.87 mM L−1 min−1, respectively. In vitro studies showed that His-MDH exhibited maximal MDH activity in pH 6.0 reaction buffer at 40°C. The enzymatic activity was 100%, 60%, and 40% inhibited by Cu2+, Zn2+, and Pb2+, respectively. In addition, six amino acids in the MDH were mutated to investigate their roles in the enzymatic activity. The results showed that the substitutions of amino acids Arg 89, Asp 149, Arg 152, His 176, or Thr 231 almost abolished the activity of His-MDH. The present study will help to understand MDH’s roles in B. abortus metabolism.

  15. Enzymatic activity analysis and catalytic essential residues identification of Brucella abortus malate dehydrogenase.

    Science.gov (United States)

    Han, Xiangan; Tong, Yongliang; Tian, Mingxing; Zhang, Yuxi; Sun, Xiaoqing; Wang, Shaohui; Qiu, Xusheng; Ding, Chan; Yu, Shengqing

    2014-01-01

    Malate dehydrogenase (MDH) plays important metabolic roles in bacteria. In this study, the recombinant MDH protein (His-MDH) of Brucella abortus was purified and its ability to catalyze the conversion of oxaloacetate (OAA) to L-malate (hereon referred to as MDH activity) was analyzed. Michaelis Constant (Km) and Maximum Reaction Velocity (Vmax) of the reaction were determined to be 6.45 × 10(-3) M and 0.87 mM L(-1)min(-1), respectively. In vitro studies showed that His-MDH exhibited maximal MDH activity in pH 6.0 reaction buffer at 40°C. The enzymatic activity was 100%, 60%, and 40% inhibited by Cu(2+), Zn(2+), and Pb(2+), respectively. In addition, six amino acids in the MDH were mutated to investigate their roles in the enzymatic activity. The results showed that the substitutions of amino acids Arg 89, Asp 149, Arg 152, His 176, or Thr 231 almost abolished the activity of His-MDH. The present study will help to understand MDH's roles in B. abortus metabolism.

  16. Intratracheal infection as an efficient route for testing vaccines against Chlamydia abortus in sheep.

    Science.gov (United States)

    Álvarez, D; Salinas, J; Buendía, A J; Ortega, N; del Río, L; Sánchez, J; Navarro, J A; Gallego, M C; Murcia-Belmonte, A; Cuello, F; Caro, M R

    2015-09-01

    Pregnant ewes have been widely used to test vaccines against Chlamydia abortus. However, this model entails many disadvantages such as high economic costs and long periods of pregnancy. The murine model is very useful for specific studies but cannot replace the natural host for the later stages of vaccine evaluation. Therefore, a non-pregnant model of the natural host might be useful for a vaccine trial to select the best vaccine candidates prior to use of the pregnant model. With this aim, two routes of infection were assessed in young non-pregnant sheep, namely, intranasal (IN) and intratracheal (IT). In addition, groups of non-vaccinated sheep and sheep immunised with an inactivated vaccine were established to investigate the suitability of the model for testing vaccines. After the experimental infection, isolation of the microorganism in several organs, with pathological and immunohistochemical analyses, antibody production assessment and investigation by PCR of the presence of chlamydia in the vagina or rectum were carried out. Experimental IT inoculation of C. abortus induced pneumonia in sheep during the first few days post-infection, confirming the suitability of the IT route for testing vaccines in the natural host. The course of infection and the resulting pathological signs were less severe in vaccinated sheep compared with non-vaccinated animals, demonstrating the success of vaccination. IN infection did not produce evident lesions or demonstrate the presence of chlamydial antigen in the lungs and cannot be considered an appropriate model for testing vaccines.

  17. An evaluation of ELISA using recombinant Brucella abortus bacterioferritin (Bfr) for bovine brucellosis.

    Science.gov (United States)

    Hop, Huynh Tan; Arayan, Lauren Togonon; Simborio, Hannah Leah; Reyes, Alisha Wehdnesday Bernardo; Min, WonGi; Lee, Hu Jang; Lee, Jin Ju; Chang, Hong Hee; Kim, Suk

    2016-04-01

    To date, detection of antibodies against the lipopolysaccharide portion is the backbone of most serodiagnostic methods for brucellosis screening. However this pose a risk for false positive reactions related to other pathogens especially that of Yersinia enterocolitica O:9 which has the most prominent cross reactivity with Brucella spp. In this study, cloning and expression of Brucella abortus bacterioferritin (Bfr) was accomplished by PCR amplification into an expression vector system, and purification of a recombinant B. abortus Bfr (rBfr). The immunogenicity of rBfr was confirmed by Western blot with Brucella-positive bovine serum. To determine whether rBfr has a potential benefit for use in the serodiagnosis of bovine brucellosis, rBfr-based ELISA was performed. Interestingly, rBfr was able to detect anti-Brucella antibodies in positive sera in a dependent manner of TAT values but did not show an immunoreaction with negative samples. Particularly, average OD492 values at the lowest, medium and highest TAT titer levels were 1.4, 2.2 and 2.6-fold increase compared with the cutoff value, respectively. The accuracy, specificity and sensitivity of rBfr showed 89.09%, 93.6% and 85.33%, respectively. These findings suggest that rBfr might be a good candidate for serological diagnosis development of bovine brucellosis.

  18. Seroprevalence and molecular characterization of Chlamydia abortus in frozen fetal and placental tissues of aborting ewes in northeastern Algeria.

    Science.gov (United States)

    Hireche, Sana; Ababneh, Mustafa Mohammed Kheir; Bouaziz, Omar; Boussena, Sabrina

    2016-02-01

    Enzootic abortion of ewes is one of the most serious health problems in sheep flocks worldwide. It has a significant economic impact because abortion, decrease in milk production and weak lambs. Besides, the bacteria is zoonotic. A cross-sectional study was conducted to determine the seroprevalence and risk factors associated with Chlamydia abortus infection in 552 ewes in Constantine using a C. abortus-specific indirect ELISA kit. Chlamydial DNA was investigated in ten ovine fetuses and eight placentas using PCR- restriction fragment length polymorphism (RFLP) and DNA sequencing. The study concluded that 7.2 % of ewes were seropositive and 33.3 % of sheep flocks had at least one seropositive ewe. Adjacent farmworker visits (OR = 7.667, 95 % CI (OR) = 2.307; 27.203) was defined as a risk factor. Deliveries of weak lambs (OR = 2.920, 95 % CI (OR) = 1.022; 8.342) and septicemia in lambs (OR = 9.971, 95 % CI (OR) = 2.383; 41.713) were significantly associated with chlamydial infection. PCR-RFLP analysis revealed positive signals to C. abortus in six fetuses and four placentas. Sequencing of the omp2 gene revealed that the Algerian strain is 96 % similar with C. abortus FAS strain. C. abortus plays a major role in abortion in northeastern Algeria. Appropriate control measures must be implemented to reduce economic losses and to avoid human contamination.

  19. Infection of C57BL/6 mice by Trypanosoma musculi modulates host immune responses during Brucella abortus cocolonization.

    Science.gov (United States)

    Lowry, Jake E; Leonhardt, Jack A; Yao, Chaoqun; Belden, E Lee; Andrews, Gerard P

    2014-01-01

    Brucellosis, which results in fetal abortions in domestic and wildlife animal populations, is of major concern in the US and throughout much of the world. The disease, caused by Brucella abortus, poses an economic threat to agriculture-based communities. A moderately efficacious live attenuated vaccine (B. abortus strain RB51) exists. However, even with vaccine use, outbreaks occur. Evidence suggests that elk (Cervus canadensis), a wild host reservoir, are the source of recent outbreaks in domestic cattle herds in Wyoming, USA. Brucella abortus establishes a chronic, persistent infection in elk. The molecular mechanisms allowing the establishment of this persistent infective state are currently unknown. A potential mechanism could be that concurrent pathogen burdens contribute to persistence. In Wyoming, elk are chronically infected with Trypanosoma cervi, which may modulate host responses in a similar manner to that documented for other trypanosomes. To identify any synergistic relationship between the two pathogens, we simulated coinfection in the well-established murine brucellosis model using Trypanosoma musculi and B. abortus S19. Groups of C57BL/6 mice (Mus musculus) were infected with either B. abortus strain 19 (S19) or T. musculi or both. Sera were collected weekly; spleens from euthanized mice were tested to determine bacterial load near the end of normal brucellosis infection. Although changes in bacterial load were observed during the later stages of brucellosis in those mice coinfected with T. musculi, the most significant finding was the suppression of gamma interferon early during the infection along with an increase in interleukin-10 secretion compared with mice infected with either pathogen alone. These results suggest that immune modulatory events occur in the mouse during coinfection and that further experiments are warranted to determine if T. cervi impacts Brucella infection in elk.

  20. Prevalence of Brucella abortus antibodies in equines of a tropical region of Mexico

    OpenAIRE

    Acosta-González, Rosa I.; González-Reyes, Ismael; Flores-Gutiérrez, Gerardo H.

    2006-01-01

    A cross-sectional study was conducted to determinate the seroprevalence rate of equine brucellosis in the state of Tamaulipas, Mexico. Serum samples from 420 equines were analyzed with the Rose Bengal test at cell concentrations of 3% (RBT-3%) and 8% (RBT-8%), and positive results were confirmed with the Rivanol test (RT). Risk factors were determined with the prevalence ratio (PR) and the use of variables generated from a questionnaire administered to the animals’ owners. Serum from 1 stalli...

  1. Prevalence of Brucella abortus antibodies in equines of a tropical region of Mexico.

    Science.gov (United States)

    Acosta-González, Rosa I; González-Reyes, Ismael; Flores-Gutiérrez, Gerardo H

    2006-10-01

    A cross-sectional study was conducted to determinate the seroprevalence rate of equine brucellosis in the state of Tamaulipas, Mexico. Serum samples from 420 equines were analyzed with the Rose Bengal test at cell concentrations of 3% (RBT-3%) and 8% (RBT-8%), and positive results were confirmed with the Rivanol test (RT). Risk factors were determined with the prevalence ratio (PR) and the use of variables generated from a questionnaire administered to the animals' owners. Serum from 1 stallion had positive results with both the RBT-8% and the RT, for a seroprevalence rate of 0.238%. Drinking of water from a pond that was also used by cattle and dogs was the only associated risk factor for this animal (PR = 0.25). However, the results were considered false-positive, because the results for other horses in the same environmental conditions were negative. Although brucellosis is considered endemic in ruminants in the study area, the results obtained suggest that equines are not a reservoir of brucellosis and do not play an important role in the epidemiologic patterns of this disease in northeastern Mexico.

  2. TLR9 is required for MAPK/NF-κB activation but does not cooperate with TLR2 or TLR6 to induce host resistance to Brucella abortus.

    Science.gov (United States)

    Gomes, Marco Túlio; Campos, Priscila Carneiro; Pereira, Guilherme de Sousa; Bartholomeu, Daniella Castanheira; Splitter, Gary; Oliveira, Sergio Costa

    2016-05-01

    Brucella abortus is a Gram-negative intracellular bacterial pathogen that causes a zoonosis of worldwide occurrence, leading to undulant fever in humans and abortion in domestic animals. B. abortus is recognized by several pattern-recognition receptors triggering pathways during the host innate immune response. Therefore, here, we determined the cooperative role of TLR9 with TLR2 or TLR6 receptors in sensing Brucella Furthermore, we deciphered the host innate immune response against B. abortus or its DNA, emphasizing the role of TLR9-MAPK/NF-κB signaling pathways in the production of proinflammatory cytokines. TLR9 is required for the initial host control of B. abortus, but this TLR was dispensable after 6 wk of infection. The susceptibility of TLR9(-/-)-infected animals to Brucella paralleled with lower levels of IFN-γ produced by mouse splenocytes stimulated with this pathogen compared with wild-type cells. However, no apparent cooperative interplay was observed between TLR2-TLR9 or TLR6-TLR9 receptors to control infection. Moreover, B. abortus or its DNA induced activation of MAPK/NF-κB pathways and production of IL-12 and TNF-α by macrophages partially dependent on TLR9 but completely dependent on MyD88. In addition, B. abortus-derived CpG oligonucleotides required TLR9 to promote IL-12 and TNF-α production by macrophages. By confocal microscopy, we demonstrated that TLR9 redistributed and colocalized with lysosomal-associated membrane protein-1 upon Brucella infection. Thus, B. abortus induced TLR9 traffic, leading to cell signaling activation and IL-12 and TNF-α production. Although TLR9 recognized Brucella CpG motifs, our results suggest a new pathway of B. abortus DNA-activating macrophages independent of TLR9.

  3. FREQÜÊNCIA DE AGLUTININAS ANTI-Brucella abortus EM CAPRINOS E OVINOS DO SERTÃO DO ESTADO DE PERNAMBUCO, BRASIL FREQUENCY OF ANTI-Brucella abortus AGGLUTININS IN GOATS AND sheep OF THE “SERTÃO” (BACKLANDS OF THE STATE OF PERNAMBUCO, BRAZIL

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    Vânia Lúcia de Assis Santana

    2008-12-01

    .936 and sex (p = 0.562. Of the 360 sheep samples, nine (2.5% were reactive. There also was no significant association between the analyzed variables and the seropositiveness for brucellosis: age group (p = 0.522; race (p = 0.576 and sex (p = 0.461. Significant association was observed (p = 0.042 among the studied species and seropositiveness for brucellosis in the investigated animals. The seropositiveness for Brucella abortus in goats and sheep was traced for the first time in the “Sertão” (dry interior region, backlands of Pernambuco, fact that can hinder the success of the National Program of Control and Erradication of Brucellosis, due to the fact that it is common to raise small ruminants with bovines in this area, besides representing risks to Public Health.
     
    KEY WORDS: Brucellosis, ovines, caprines, small ruminants, serodiagnosis.

  4. Effect of Preventive Chlamydia abortus Vaccination in Offspring Development in Sheep Challenged Experimentally

    Science.gov (United States)

    García-Seco, Teresa; Pérez-Sancho, Marta; Salinas, Jesús; Navarro, Alejandro; Díez-Guerrier, Alberto; García, Nerea; Pozo, Pilar; Goyache, Joaquín; Domínguez, Lucas; Álvarez, Julio

    2016-01-01

    Ovine enzootic abortion, caused by Chlamydia abortus, leads to important economic losses worldwide. In addition to reproductive failures, infection may impact lamb growth during the first weeks after birth, yet this effect has not been well characterized. Vaccination can help to control the disease but variable efficacy values have been described, possibly related with factors associated with the host, the vaccine, the parameter used for efficacy determination, and the challenge conditions. In this context, we evaluated the efficacy of an inactivated standard commercial vaccine and a 1/2 diluted dose in pregnant sheep challenged with C. abortus by examining multiple indicators of vaccine effect (including incidence of reproductive failures, bacterial excretion, and evolution of weight gain of viable lambs during the first month of life). Three groups of ewes [control non-vaccinated, C (n = 18); vaccinated with standard dose, SV (n = 16); and vaccinated with 1/2 dose, DV (n = 17)], were challenged approximately 90 days post-mating and tested using direct PCR (tissue samples and vaginal swabs) and ELISA (serum) until 31 days post-reproductive outcome. There were not significant differences in the proportions of reproductive failures or bacterial shedding after birth/abortion regardless the vaccination protocol. However, a beneficial effect of vaccination on offspring growth was detected in both vaccinated groups compared with the controls, with a mean increase in weight measured at 30 days of life of 1.5 and 2.5 kg (p = 0.056) and an increase in the geometric mean of the daily gain of 8.4 and 9.7% in lambs born from DV and SV ewes compared with controls, respectively. Our results demonstrate the effect of an inactivated vaccine in the development of the offspring of C. abortus-infected ewes at a standard and a diluted dose, an interesting finding given the difficulty in achieving sufficient antigen concentration in the production of enzootic

  5. Adrenal steroids modulate the immune response during Brucella abortus infection by a mechanism that depends on the regulation of cytokine production.

    Science.gov (United States)

    Gentilini, María Virginia; Velásquez, Lis Noelia; Barrionuevo, Paula; Arriola Benitez, Paula Constanza; Giambartolomei, Guillermo Hernán; Delpino, María Victoria

    2015-05-01

    Human brucellosis is a protean disease with a diversity of clinical signs and symptoms resulting from infection with Brucella species. Recent reports suggest a cross-regulation between adrenal steroids (cortisol and dehydroepiandrosterone [DHEA]) and the immune system. Monocytes and macrophages are the main replication niche for Brucella. Therefore, we investigated the role of adrenal hormones on the modulation of the immune response mediated by macrophages in B. abortus infection. Cortisol treatment during B. abortus infection significantly inhibits cytokine, chemokine, and MMP-9 secretion. In contrast, DHEA treatment had no effect. However, DHEA treatment increases the expression of costimulatory molecules (CD40, CD86), the adhesion molecule CD54, and major histocompatibility complex class I (MHC-I) and MHC-II expression on the surface of B. abortus-infected monocytes. It is known that B. abortus infection inhibits MHC-I and MHC-II expression induced by gamma interferon (IFN-γ) treatment. DHEA reverses B. abortus downmodulation of the MHC-I and -II expression induced by IFN-γ. Taken together, our data indicate that DHEA immune intervention may positively affect monocyte activity during B. abortus infection.

  6. The effects of red ginseng saponin fraction-A (RGSF-A) on phagocytosis and intracellular signaling in Brucella abortus infected RAW 264.7 cells.

    Science.gov (United States)

    Arayan, Lauren Togonon; Simborio, Hannah Leah; Reyes, Alisha Wehdnesday Bernardo; Hop, Huynh Tan; Min, WonGi; Lee, Hu Jang; Rhee, Man Hee; Chang, Hong Hee; Kim, Suk

    2015-06-01

    This study indicated that RGSF-A caused a marked reduction in the adherence, internalization and intracellular growth of Brucella abortus in RGSF-A-treated cells. Furthermore, a decline in the intensity of F-actin fluorescence was observed in RGSF-A-treated cells compared with untreated B. abortus-infected cells. In addition, an evaluation of phagocytic signaling proteins by Western blot analysis revealed an apparent reduction of ERK and p38α phosphorylation levels in B. abortus-infected RGSF-A-treated cells compared with the control. Upon intracellular trafficking of the pathogen, a higher number of B. abortus-containing phagosomes colocalized with LAMP-1 in RGSF-A-treated cells compared with control cells. These results strongly suggest that inhibition of B. abortus uptake could be mediated by suppression in the activation of MAPKs signaling proteins phospho-ERK 1/2, and p38 levels. On the other hand, inhibition of intracellular replication results from the enhancement of phagolysosome fusion in host macrophages. This study highlights the phagocytic and intracellular modulating effect of RGSF-A and its potential as an alternative remedy to control B. abortus infection.

  7. Protective immune-response of aluminium hydroxide gel adjuvanted phage lysate of Brucella abortus S19 in mice against direct virulent challenge with B. abortus 544.

    Science.gov (United States)

    Jain, Lata; Rawat, Mayank; Prajapati, Awadhesh; Tiwari, Ashok Kumar; Kumar, Bablu; Chaturvedi, V K; Saxena, H M; Ramakrishnan, Sarvanan; Kumar, Jatin; Kerketta, Priscilla

    2015-09-01

    The prophylactic efficacies of plain and alum adsorbed lysate were evaluated by direct virulent challenge in mice model. A recently isolated brucellaphage 'ϕLd' was used for generation of lysates. Twenty four h incubated Brucella abortus S19 broth cultures standardized to contain approximately 10(8) CFU/ml were found suitable for generation of lysates. Three lysate batches produced through separate cycles did not show any significant variation with respect to protein and polysaccharide contents, endotoxin level and phage counts, indicating that compositionally stable lysate preparations can be generated through an optimized production process. Three polypeptides of ∼16, 19 and 23 kDa could be identified as immuno-dominant antigens of the lysate which induced both humoral and cell-mediated immune responses in a dose dependent manner. Results of efficacy evaluation trial confirmed dose-dependent protective potencies of lysate preparation. The lysate with an antigenic dose of 0.52 μg protein and 60 μg CHO adsorbed on aluminium gel (0.1 percent aluminium concentration) exhibited the highest protective potency which was greater than that induced by standard S19 vaccine. Phage lysate methodology provides a very viable option through which an improved immunizing preparation with all desirable traits can be developed against brucellosis, and integrated with immunization programmes in a more efficient manner.

  8. Medición de respuesta inmune humoral y celular frente a antígenos de Brucella abortus RB51 en bovinos (Evaluation of Humoral and cellular immune response evaluation against Brucella abortus strain RB51 antigens in bovine

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    N.I. Montaña S.

    1998-01-01

    Full Text Available Con el objeto de evaluar comparativamente el tipo de respuesta inmune inducido por los antígenos estructurales purificados y vacunas vivas de Brucella abortus, 14 bovinos criollos hembras de 19 meses de edad fueron distribuidos al azar, en cinco grupos experimentales e inmunizados vía subcutánea de la siguiente manera: OMP-II (proteínas de membrana externa, OMP-II-Cadena-O, OMP II acoplado a polisacárido-O, B. abortus cepa RB51, B. abortus cepa 19 (C19 y solución salina para el grupo control. Dos meses posinmunización, los animales fueron desafiados, vía intramuscular, con cepa patógena de referencia B. abortus 2308 y evaluados en su nivel de protección 30 días posdesafío. La respuesta inmune humoral específica fue determinada mediante las pruebas convencionales, de aglutinación Rosa de Bengala, fijación de complemento, inmunodifusión radial, ELISA indirecta, a los 8, 15, 30, 60 y 90 días posinmunización. Adicionalmente se determinó la subclase de IgG por ELISA tipo doble sandwich. La evaluación de respuesta celular fue determinada mediante linfoproliferación, expresada en índices de estimulación (IS, obtenidos por los niveles de incorporación de 3H timidina, medición de Interferón gamma (IFN-g, igualmente expresado como IS y cuantificación de subpoblaciones linfoides CD4+ y CD8+ por citofluorometría. Animales vacunados con cepas vivas demostraron protección total al reto, mientras se observó un 66% de protección en los vacunados con antígenos purificados. Se observó ausencia de anticuerpos a lipopolisacárido, sLPS, por las pruebas convencionales para B. abortus en todos los grupos inmunizados, excepto el vacunado con cepa 19, confirmando la ventaja diagnóstica de la utilización de la cepa RB51 o sus antígenos purificados. La linfoproliferación como una de las medidas de respuesta celular no demostró IS significativos (p0.05 frente a los antígenos estructurales purificados, OMP-II, Cadena-O y sLPS en

  9. Brucella abortus Invasion of Osteocytes Modulates Connexin 43 and Integrin Expression and Induces Osteoclastogenesis via Receptor Activator of NF-κB Ligand and Tumor Necrosis Factor Alpha Secretion.

    Science.gov (United States)

    Pesce Viglietti, Ayelén Ivana; Arriola Benitez, Paula Constanza; Gentilini, María Virginia; Velásquez, Lis Noelia; Fossati, Carlos Alberto; Giambartolomei, Guillermo Hernán; Delpino, María Victoria

    2015-10-12

    Osteoarticular brucellosis is the most common localization of human active disease. Osteocytes are the most abundant cells of bone. They secrete factors that regulate the differentiation of both osteoblasts and osteoclasts during bone remodeling. The aim of this study is to determine if Brucella abortus infection modifies osteocyte function. Our results indicate that B. abortus infection induced matrix metalloproteinase 2 (MMP-2), receptor activator for NF-κB ligand (RANKL), proinflammatory cytokines, and keratinocyte chemoattractant (KC) secretion by osteocytes. In addition, supernatants from B. abortus-infected osteocytes induced bone marrow-derived monocytes (BMM) to undergo osteoclastogenesis. Using neutralizing antibodies against tumor necrosis factor alpha (TNF-α) or osteoprotegerin (OPG), RANKL's decoy receptor, we determined that TNF-α and RANKL are involved in osteoclastogenesis induced by supernatants from B. abortus-infected osteocytes. Connexin 43 (Cx43) and the integrins E11/gp38, integrin-α, integrin-β, and CD44 are involved in cell-cell interactions necessary for osteocyte survival. B. abortus infection inhibited the expression of Cx43 but did not modify the expression of integrins. Yet the expression of both Cx43 and integrins was inhibited by supernatants from B. abortus-infected macrophages. B. abortus infection was not capable of inducing osteocyte apoptosis. However, supernatants from B. abortus-infected macrophages induced osteocyte apoptosis in a dose-dependent manner. Taken together, our results indicate that B. abortus infection could alter osteocyte function, contributing to bone damage.

  10. Detecção de Brucella abortus em tecidos bovinos utilizando ensaios de PCR e qPCR¹

    Directory of Open Access Journals (Sweden)

    Marrielen A.B. Caitano

    2014-06-01

    Full Text Available Objetivou-se no presente estudo avaliar as técnicas reação em cadeia da polimerase (PCR e PCR em Tempo Real (qPCR para detectar Brucella abortus, a partir de tecidos bovinos com lesões sugestivas de brucelose. Para isto, 21 fragmentos de tecidos bovinos coletados em abatedouros de Mato Grosso do Sul foram processados e submetidos ao cultivo microbiológico e extração do DNA genômico para realização das reações de PCR e qPCR. No cultivo microbiológico, oito amostras apresentaram crescimento bacteriano e cinco foram confirmadas como B. abortus por PCR. Diretamente das amostras de tecido, DNA do gênero Brucella (oligonucleotídeos IS711 foi detectado em 13 (61,9% amostras de tecido e 17 (81% amostras de homogeneizado. Já com os oligonucleotídeos espécie-específicos BruAb2_0168F e BruAb2_0168R, 14 (66% amostras de tecido e 18 (85,7% amostras de homogeneizado foram amplificadas. Seis amostras positivas na PCR espécie-específica foram sequenciadas e o best hit na análise BLASTn foi B. abortus. Na qPCR, 21 (100% amostras de tecidos e 19 (90,5% amostras de homogeneizado foram positivas para B. abortus. Dez amostras de DNA de sangue bovino de rebanho certificado livre foram utilizadas como controle negativo nas análises de PCR e qPCR utilizando-se os oligonucleotídeos BruAb2_0168F e BruAb2_0168R. Na PCR nenhuma amostra amplificou, enquanto que na qPCR 2 (20% amplificaram. Conclui-se que as duas técnicas detectam a presença de B. abortus diretamente de tecidos e homogeneizados, porém a qPCR apresentou maior sensibilidade. Os resultados obtidos indicam que a qPCR pode representar uma alternativa rápida e precisa para a detecção de B. abortus diretamente de tecidos, e ser utilizada em programas de vigilância sanitária, por apresentar sensibilidade e especificidade satisfatórias.

  11. Brucella abortus S19 vaccine protects dairy cattle against natural infection with Brucella melitensis.

    Science.gov (United States)

    van Straten, Michael; Bardenstein, Svetlana; Keningswald, Gaby; Banai, Menachem

    2016-11-21

    Brucellosis is a zoonotic disease that can cause severe illness in humans and considerable economic loss in the livestock industry. Although small ruminants are the preferential host for Brucella melitensis, this pathogen has emerged as a cause for Brucella outbreaks in cattle. S19 vaccination is implemented in many countries where B. abortus is endemic but its effectiveness against B. melitensis has not been validated. Here we show that vaccine effectiveness in preventing disease transmission between vaccinated and unvaccinated cohorts, as determined by seroconversion, was 87.2% (95% CI 69.5-94.6%). Furthermore, vaccination was associated with a reduced risk for abortion. Together, our data emphasize the role S19 vaccination could play in preventing B. melitensis outbreaks in areas where this pathogen is prevalent in small ruminant populations.

  12. Milk Ring Test for spot identification of Brucella abortus infection in single cow herds

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    Najibullah Mohamand

    2014-06-01

    Full Text Available In this study, milk samples were collected from 109 dairy cows to detect antibodies against Brucella (B. using Milk Ring Test (MRT. Overall, 18.35% (n=20/109 of the milk samples were positive by MRT. The cows were divided into three groups based on lactation number viz., 1st, 2nd to 4th and ≥5th lactations; the prevalence of brucellosis in the groups were found to be 0.92% (n=1/109, 15.60% (n=17/109 and 1.83% (n=2/109, respectively. Considering simplicity and cost effectiveness, the MRT can be used for the preliminary screening of B. abortus infection especially in single cow herds.

  13. Brucella abortus Strain 2308 Wisconsin Genome: Importance of the Definition of Reference Strains

    Science.gov (United States)

    Suárez-Esquivel, Marcela; Ruiz-Villalobos, Nazareth; Castillo-Zeledón, Amanda; Jiménez-Rojas, César; Roop II, R. Martin; Comerci, Diego J.; Barquero-Calvo, Elías; Chacón-Díaz, Carlos; Caswell, Clayton C.; Baker, Kate S.; Chaves-Olarte, Esteban; Thomson, Nicholas R.; Moreno, Edgardo; Letesson, Jean J.; De Bolle, Xavier; Guzmán-Verri, Caterina

    2016-01-01

    Brucellosis is a bacterial infectious disease affecting a wide range of mammals and a neglected zoonosis caused by species of the genetically homogenous genus Brucella. As in most studies on bacterial diseases, research in brucellosis is carried out by using reference strains as canonical models to understand the mechanisms underlying host pathogen interactions. We performed whole genome sequencing analysis of the reference strain B. abortus 2308 routinely used in our laboratory, including manual curated annotation accessible as an editable version through a link at https://en.wikipedia.org/wiki/Brucella#Genomics. Comparison of this genome with two publically available 2308 genomes showed significant differences, particularly indels related to insertional elements, suggesting variability related to the transposition of these elements within the same strain. Considering the outcome of high resolution genomic techniques in the bacteriology field, the conventional concept of strain definition needs to be revised. PMID:27746773

  14. Evaluation of Brucella abortus strain RB51 and strain 19 in pronghorn antelope

    Science.gov (United States)

    Elzer, P.H.; Smith, J.; Roffe, T.; Kreeger, T.; Edwards, J.; Davis, D.

    2002-01-01

    Free-roaming elk and bison in the Greater Yellowstone Area remain the only wildlife reservoirs for Brucella abortus in the United States, and the large number of animals and a lack of holding facilities make it unreasonable to individually vaccinate each animal. Therefore, oral delivery is being proposed as a possible option to vaccinate these wild ungulates. One of the main problems associated with oral vaccination is the potential exposure of nontarget species to the vaccines. The purpose of this study was to determine the effects of two Brucella vaccines, strain 19 (S19) and the rough strain RB51 (SRB51), in pregnant pronghorn antelope. We conclude that S19 and SRB51 rarely colonize maternal and fetal tissues of pregnant pronghorn and were not associated with fetal death. Oral delivery of either vaccine at this dose appears to be nonhazardous to pregnant pronghorn.

  15. First report of orchitis in man caused by Brucella abortus biovar 1 in Ecuador.

    Science.gov (United States)

    Ron-Román, Jorge; Saegerman, Claude; Minda-Aluisa, Elizabeth; Benítez-Ortíz, Washington; Brandt, Jef; Douce, Richard

    2012-09-01

    We present a 44-year-old man from a rural community in northern Ecuador who worked on a cattle farm where he was involved with primary veterinary care, including assistance during births (or calving) and placenta retention and artificial insemination, with minimal precautions. In September of 2009, quite abruptly, he developed asthenia and hypersomnia without any apparent cause or symptoms like fever, chills, or night sweats. On November 14, 2009, he suffered from pain and edema in the right testicle that coincided with pain in the abdomen. Clinical, serological, and bacteriological investigations confirmed the first case of unilateral orchitis in man in Ecuador caused by Brucella abortus biovar 1. Because brucellosis is a neglected disease, special attention should be given to it in the training of medical and veterinary students.

  16. Serology for Brucella abortus in cart horses from an urban area in Brazil

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    J.M.A.P. Antunes

    2013-04-01

    Full Text Available Avaliou-se a infecção por Brucella abortus em cavalos de carroça de Curitiba e São José dos Pinhais-PR. Um total de 123 amostras foi submetido ao teste do antígeno tamponado acidificado (ATA, soroaglutinação lenta em tubos (SAL e prova do 2-mercaptoetanol (2-ME para confirmação dos resultados. Oito (6,5% equinos foram positivos para o ATA e um animal permaneceu positivo ao teste confirmatório. Existem evidências da presença de brucelose entre os cavalos de carroça.

  17. Increasing effect of a high dose of PG-1 peptide on the infectivity of Chlamydophila abortus.

    Science.gov (United States)

    Donati, Manuela; Di Francesco, Antonietta; Gennaro, Renato; Benincasa, Monica; Di Paolo, Maria; Shurdhi, Alisa; Ostanello, Fabio; Baldelli, Raffaella; Cevenini, Roberto

    2010-07-01

    Cathelicidins are antimicrobial peptides, stored by mammalian leukocytes, showing an antimicrobial activity against bacteria, fungi, protozoa and enveloped viruses. In accordance with other authors, we reported in a previous study that the protegrin-1 (PG-1), at 80 microg mL(-1), inhibited the in vitro growth of Chlamydia trachomatis serovars D, H and L2; however, we observed an increased infectivity of some animal chlamydial species after their treatment with the same PG-1 concentration. In this study, the treatment of LLC-MK2 cells with PG-1 before chlamydial infection resulted in an increased infectivity of Chlamydophila abortus probably due to their easier entry into the host cells, whereas no increase in S26/3 infectivity was detected in LLC-MK2 cells treated with PG-1 postchlamydial infection.

  18. Crystal structure of cyclic nucleotide-binding-like protein from Brucella abortus.

    Science.gov (United States)

    He, Zheng; Gao, Yuan; Dong, Jing; Ke, Yuehua; Li, Xuemei; Chen, Zeliang; Zhang, Xuejun C

    2015-12-25

    The cyclic nucleotide-binding (CNB)-like protein (CNB-L) from Brucella abortus shares sequence homology with CNB domain-containing proteins. We determined the crystal structure of CNB-L at 2.0 Å resolution in the absence of its C-terminal helix and nucleotide. The 3D structure of CNB-L is in a two-fold symmetric form. Each protomer shows high structure similarity to that of cGMP-binding domain-containing proteins, and likely mimics their nucleotide-free conformation. A key residue, Glu17, mediates the dimerization and prevents binding of cNMP to the canonical ligand-pocket. The structurally observed dimer of CNB-L is stable in solution, and thus is likely to be biologically relevant.

  19. Efficacy of single calfhood vaccination of elk with Brucella abortus strain 19

    Science.gov (United States)

    Roffe, T.J.; Jones, L.C.; Coffin, K.; Drew, M.L.; Sweeney, Steven J.; Hagius, S.D.; Elzer, P.H.; Davis, D.

    2004-01-01

    Brucellosis has been eradicated from cattle in the states of Wyoming, Montana, and Idaho, USA. However, free-ranging elk (Cervus elaphus) that use feedgrounds in the Greater Yellowstone Area (GYA) and bison (Bison bison) in Yellowstone and Grand Teton national parks still have high seroprevalence to the disease and have caused loss of brucellosis-free status in Wyoming. Management tools to control or eliminate the disease are limited; however, wildlife vaccination is among the methods currently used by wildlife managers in Wyoming. We conducted a controlled challenge study of single calfhood vaccination. Elk calves, caught in January and February of 1999 and 2000 and acclimated to captivity for 3 weeks, were randomly assigned to control or vaccinate groups. The vaccinate groups received Brucetta abortus vaccine strain 19 (S19) by hand-delivered intramuscular injection. Calves were raised to adulthood and bred at either 2.5 or 3.5 years of age for 2000 and 1999 captures, respectively. Eighty-nine (44 controls, 45 vaccinates) pregnant elk entered the challenge portion of the study. We challenged elk at mid-gestation with pathogenic B. abortus strain 2308 by intraconjunctival instillation. Abortion occurred in significantly more (P = 0.002) controls (42; 93%) than vaccinates (32; 71%), and vaccine protected 25% of the vaccinate group. We used Brucella culture of fetus/calf tissues to determine the efficacy of vaccination for preventing infection, and we found that the number of infected fetuses/calves did not differ between controls and vaccinates (P = 0.14). Based on these data, single calfhood vaccination with S19 has low efficacy, will likely have only little to moderate effect on Brucella prevalence in elk, and is unlikely to eradicate the disease in wildlife of the GYA.

  20. DETECCION DE Brucella abortus POR PCR EN MUESTRAS DE SANGRE Y LECHE DE VACUNOS

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    Xiomara Mosquera C

    2008-12-01

    Full Text Available Objetivo. Evaluar el uso de la Reacción en Cadena de la Polimerasa (PCR para la detección de Brucella abortus en muestras de sangre y leche de vacunos. Materiales y métodos. Este estudio de tipo descriptivo fue realizado durante los años 2004 y 2005. Se analizaron 136 animales de tres fincas localizadas en el municipio de Durania, Norte de Santander, Colombia. Se evaluó la presencia de anticuerpos en la leche mediante la prueba del anillo (PAL. Se amplificó el fragmento de 223pb del gen BCSP31. Se emplearon los cebadores B4 y B5 de la región interna de la secuencia del gen BCSP31 (GenBank, número M20404. Resultados. En aquellos animales positivos se obtuvo una muestra de sangre y leche para el análisis por PCR, la sangre no fue analizada por serología. Se evaluaron diferentes métodos de extracción de ADN. Se encontró que un 13.2% (18/136 de las muestras de leche fueron positivas a la PAL. Se analizaron 33 muestras de leche negativas por PAL de las cuales el 30.3% (10/33 resultaron positivas por PCR. Al analizar las muestras de sangre de los animales positivos por PAL el 94.1% (16/17 fueron positivas por PCR, mientras que el 47% (8/17 de las muestras de leche positivas por PAL, fueron positivas por PCR. Conclusiones. Se demostró la amplificación de un fragmento de ADN de Brucella abortus en muestras de sangre y leche de vacunos. Los resultados preliminares demostraron que es posible usar PCR como prueba diagnóstica de brucelosis en Colombia.

  1. The role of NLRP3 and AIM2 in inflammasome activation during Brucella abortus infection

    Science.gov (United States)

    Gomes, Marco Tulio R.; Miraglia, Maria Cruz; Giambartolomei, Guillermo H.; Oliveira, Sergio C.

    2016-01-01

    The innate immune system is essential for detection and elimination of bacterial pathogens. Upon inflammasome activation, caspase-1 cleaves pro-IL-1β and pro-IL-18 to their mature forms IL-1β and IL-18, respectively, and the cell undergoes inflammatory death termed pyroptosis. Here we reviewed recent findings demonstrating that Brucella abortus ligands activate NLRP3 and AIM2 inflammasomes which leads to control of infection. This protective effect is due to inflammatory response caused by IL-1β and IL-18 rather than cell death. Brucella DNA is sensed by AIM2 and bacteria induced mitochondrial reactive oxygen species is detected by NLRP3. However, deregulation of proinflammatory cytokine production can lead to immunopathology. Nervous system invasion by bacteria of the genus Brucella results in an inflammatory disorder termed neurobrucellosis. Herein we discuss the mechanism of caspase-1 activation and IL-1β secretion in glial cells infected with B. abortus. Our results demonstrate that the ASC inflammasome is indispensable for inducing the activation of caspase-1 and secretion of IL-1β upon infection of astrocytes and microglia with Brucella. Moreover, our results demonstrate that secretion of IL-1β by Brucella-infected glial cells depends on NLRP3 and AIM2 and leads to neurobrucellosis. Further, the inhibition of the host cell inflammasome as an immune evasion strategy has been described for bacterial pathogens. We discuss here that the bacterial type IV secretion system VirB is required for inflammasome activation in host cells during infection. Taken together, our results indicate that Brucella is sensed by ASC inflammasomes mainly NLRP3 and AIM2 that collectively orchestrate a robust caspase-1 activation and proinflammatory response. PMID:27405866

  2. Transcriptome analysis of the Brucella abortus BvrR/BvrS two-component regulatory system.

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    Cristina Viadas

    Full Text Available BACKGROUND: The two-component BvrR/BvrS system is essential for Brucella abortus virulence. It was shown previously that its dysfunction alters the expression of some major outer membrane proteins and the pattern of lipid A acylation. To determine the genes regulated by BvrR/BvrS, we performed a whole-genome microarray analysis using B. abortus RNA obtained from wild type and bvrR mutant cells grown in the same conditions. METHODOLOGY/PRINCIPAL FINDINGS: A total of 127 differentially expressed genes were found: 83 were over expressed and 44 were less expressed in the bvrR mutant. Two operons, the phosphotransferase system and the maltose transport system, were down-regulated. Several genes involved in cell envelope or outer membrane biogenesis were differentially expressed: genes for outer membrane proteins (omp25a, omp25d, lipoproteins, LPS and fatty acid biosynthesis, stress response proteins, chaperones, flagellar genes, and twelve genes encoding ABC transport systems. Ten genes related with carbon metabolism (pckA and fumB among others were up-regulated in the bvrR mutant, and denitrification genes (nirK, norC and nosZ were also regulated. Notably, seven transcriptional regulators were affected, including VjbR, ExoR and OmpR that were less expressed in the bvrR mutant. Finally, the expression of eleven genes which have been previously related with Brucella virulence was also altered. CONCLUSIONS/SIGNIFICANCE: All these data corroborate the impact of BvrR/BvrS on cell envelope modulation, confirm that this system controls the carbon and nitrogen metabolism, and suggest a cross-talk among some regulators to adjust the Brucella physiology to the shift expected to occur during the transit from the extracellular to the intracellular niche.

  3. Mutant Brucella abortus membrane fusogenic protein induces protection against challenge infection in mice.

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    de Souza Filho, Job Alves; de Paulo Martins, Vicente; Campos, Priscila Carneiro; Alves-Silva, Juliana; Santos, Nathalia V; de Oliveira, Fernanda Souza; Menezes, Gustavo B; Azevedo, Vasco; Cravero, Silvio Lorenzo; Oliveira, Sergio Costa

    2015-04-01

    Brucella species can cause brucellosis, a zoonotic disease that causes serious livestock economic losses and represents a public health threat. The mechanism of virulence of Brucella spp. is not yet fully understood. Therefore, it is crucial to identify new molecules that serve as virulence factors to better understand this host-pathogen interplay. Here, we evaluated the role of the Brucella membrane fusogenic protein (Mfp) and outer membrane protein 19 (Omp19) in bacterial pathogenesis. In this study, we showed that B. abortus Δmfp::kan and Δomp19::kan deletion mutant strains have reduced persistence in vivo in C57BL/6 and interferon regulatory factor 1 (IRF-1) knockout (KO) mice. Additionally, 24 h after macrophage infection with a Δmfp::kan or Δomp19::kan strain expressing green fluorescent protein (GFP) approximately 80% or 65% of Brucella-containing vacuoles (BCVs) retained the late endosomal/lysosomal marker LAMP-1, respectively, whereas around 60% of BCVs containing wild-type S2308 were found in LAMP-1-negative compartments. B. abortus Δomp19::kan was attenuated in vivo but had a residual virulence in C57BL/6 and IRF-1 KO mice, whereas the Δmfp::kan strain had a lower virulence in these same mouse models. Furthermore, Δmfp::kan and Δomp19::kan strains were used as live vaccines. Challenge experiments revealed that in C57BL/6 and IRF-1 KO mice, the Δmfp::kan strain induced greater protection than the vaccine RB51 and protection similar that of vaccine S19. However, a Δomp19::kan strain induced protection similar to that of RB51. Thus, these results demonstrate that Brucella Mfp and Omp19 are critical for full bacterial virulence and that the Δmfp::kan mutant may serve as a potential vaccine candidate in future studies.

  4. CCL20 and Beta-Defensin 2 Production by Human Lung Epithelial Cells and Macrophages in Response to Brucella abortus Infection

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    Fernández, Andrea G.; Bonetto, Josefina; Giambartolomei, Guillermo H.; Fossati, Carlos A.; Baldi, Pablo C.

    2015-01-01

    Both CCL20 and human β-defensin 2 (hBD2) interact with the same membrane receptor and display chemotactic and antimicrobial activities. They are produced by airway epithelia in response to infectious agents and proinflammatory cytokines. Whereas Brucella spp. can infect humans through inhalation, their ability to induce CCL20 and hBD2 in lung cells is unknown. Here we show that B. abortus induces CCL20 expression in human alveolar (A549) or bronchial (Calu-6) epithelial cell lines, primary alveolar epithelial cells, primary human monocytes, monocyte-derived macrophages and the monocytic cell line THP-1. CCL20 expression was mainly mediated by JNK1/2 and NF-kB in both Calu-6 and THP-1 cells. CCL20 secretion was markedly induced in A549, Calu-6 and THP-1 cells by heat-killed B. abortus or a model Brucella lipoprotein (L-Omp19) but not by the B. abortus lipopolysaccharide (LPS). Accordingly, CCL20 production by B. abortus-infected cells was strongly TLR2-dependent. Whereas hBD2 expression was not induced by B. abortus infection, it was significantly induced in A549 cells by conditioned media from B. abortus-infected THP-1 monocytes (CMB). A similar inducing effect was observed on CCL20 secretion. Experiments using blocking agents revealed that IL-1β, but not TNF-α, was involved in the induction of hBD2 and CCL20 secretion by CMB. In the in vitro antimicrobial assay, the lethal dose (LD) 50 of CCL20 for B. abortus (>50 μg/ml) was markedly higher than that against E. coli (1.5 μg/ml) or a B. abortus mutant lacking the O polysaccharide in its LPS (8.7 ug/ml). hBD2 did not kill any of the B. abortus strains at the tested concentrations. These results show that human lung epithelial cells secrete CCL20 and hBD2 in response to B. abortus and/or to cytokines produced by infected monocytes. Whereas these molecules do not seem to exert antimicrobial activity against this pathogen, they could recruit immune cells to the infection site. PMID:26448160

  5. CCL20 and Beta-Defensin 2 Production by Human Lung Epithelial Cells and Macrophages in Response to Brucella abortus Infection.

    Science.gov (United States)

    Hielpos, M Soledad; Ferrero, Mariana C; Fernández, Andrea G; Bonetto, Josefina; Giambartolomei, Guillermo H; Fossati, Carlos A; Baldi, Pablo C

    2015-01-01

    Both CCL20 and human β-defensin 2 (hBD2) interact with the same membrane receptor and display chemotactic and antimicrobial activities. They are produced by airway epithelia in response to infectious agents and proinflammatory cytokines. Whereas Brucella spp. can infect humans through inhalation, their ability to induce CCL20 and hBD2 in lung cells is unknown. Here we show that B. abortus induces CCL20 expression in human alveolar (A549) or bronchial (Calu-6) epithelial cell lines, primary alveolar epithelial cells, primary human monocytes, monocyte-derived macrophages and the monocytic cell line THP-1. CCL20 expression was mainly mediated by JNK1/2 and NF-kB in both Calu-6 and THP-1 cells. CCL20 secretion was markedly induced in A549, Calu-6 and THP-1 cells by heat-killed B. abortus or a model Brucella lipoprotein (L-Omp19) but not by the B. abortus lipopolysaccharide (LPS). Accordingly, CCL20 production by B. abortus-infected cells was strongly TLR2-dependent. Whereas hBD2 expression was not induced by B. abortus infection, it was significantly induced in A549 cells by conditioned media from B. abortus-infected THP-1 monocytes (CMB). A similar inducing effect was observed on CCL20 secretion. Experiments using blocking agents revealed that IL-1β, but not TNF-α, was involved in the induction of hBD2 and CCL20 secretion by CMB. In the in vitro antimicrobial assay, the lethal dose (LD) 50 of CCL20 for B. abortus (>50 μg/ml) was markedly higher than that against E. coli (1.5 μg/ml) or a B. abortus mutant lacking the O polysaccharide in its LPS (8.7 ug/ml). hBD2 did not kill any of the B. abortus strains at the tested concentrations. These results show that human lung epithelial cells secrete CCL20 and hBD2 in response to B. abortus and/or to cytokines produced by infected monocytes. Whereas these molecules do not seem to exert antimicrobial activity against this pathogen, they could recruit immune cells to the infection site.

  6. Brucella abortusΔcydCΔcydD and ΔcydCΔpurD double-mutants are highly attenuated and confer long-term protective immunity against virulent Brucella abortus.

    Science.gov (United States)

    Truong, Quang Lam; Cho, Youngjae; Park, Soyeon; Kim, Kiju; Hahn, Tae-Wook

    2016-01-04

    We constructed double deletion (ΔcydCΔcydD and ΔcydCΔpurD) mutants from virulent Brucella abortus biovar 1 field isolate (BA15) by deleting the genes encoding an ATP-binding cassette-type transporter (cydC and cydD genes) and a phosphoribosylamine-glycine ligase (purD). Both BA15ΔcydCΔcydD and BA15ΔcydCΔpurD double-mutants exhibited significant attenuation of virulence when assayed in murine macrophages or in BALB/c mice. Both double-mutants were readily cleared from spleens by 4 weeks post-inoculation even when inoculated at the dose of 10(8) CFU per mouse. Moreover, the inoculated mice showed no splenomegaly, which indicates that the mutants are highly attenuated. Importantly, the attenuation of in vitro and in vivo growth did not impair the ability of these mutants to confer long-term protective immunity in mice against challenge with B. abortus strain 2308. Vaccination of mice with either mutant induced humoral and cell-mediated immune responses, and provided significantly better protection than commercial B. abortus strain RB51 vaccine. These results suggest that highly attenuated BA15ΔcydCΔcydD and BA15ΔcydCΔpurD mutants can be used effectively as potential live vaccine candidates against bovine brucellosis.

  7. Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts

    OpenAIRE

    Gamal Wareth; Murat Eravci; Christoph Weise; Uwe Roesler; Falk Melzer; Sprague, Lisa D.; Heinrich Neubauer; Jayaseelan Murugaiyan

    2016-01-01

    Brucellosis is a debilitating zoonotic disease that affects humans and animals. The diagnosis of brucellosis is challenging, as accurate species level identification is not possible with any of the currently available serology-based diagnostic methods. The present study aimed at identifying Brucella (B.) species-specific proteins from the closely related species B. abortus and B. melitensis using sera collected from naturally infected host species. Unlike earlier reported investigations with ...

  8. The alkylation response protein AidB is localized at the new poles and constriction sites in Brucella abortus

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    Dotreppe Delphine

    2011-11-01

    Full Text Available Abstract Background Brucella abortus is the etiological agent of a worldwide zoonosis called brucellosis. This alpha-proteobacterium is dividing asymmetrically, and PdhS, an essential histidine kinase, was reported to be an old pole marker. Results We were interested to identify functions that could be recruited to bacterial poles. The Brucella ORFeome, a collection of cloned predicted coding sequences, was placed in fusion with yellow fluorescent protein (YFP coding sequence and screened for polar localizations in B. abortus. We report that AidB-YFP was systematically localized to the new poles and at constrictions sites in B. abortus, either in culture or inside infected HeLa cells or RAW264.7 macrophages. AidB is an acyl-CoA dehydrogenase (ACAD homolog, similar to E. coli AidB, an enzyme putatively involved in destroying alkylating agents. Accordingly, a B. abortus aidB mutant is more sensitive than the wild-type strain to the lethality induced by methanesulphonic acid ethyl ester (EMS. The exposure to EMS led to a very low frequency of constriction events, suggesting that cell cycle is blocked during alkylation damage. The localization of AidB-YFP at the new poles and at constriction sites seems to be specific for this ACAD homolog since two other ACAD homologs fused to YFP did not show specific localization. The overexpression of aidB, but not the two other ACAD coding sequences, leads to multiple morphological defects. Conclusions Data reported here suggest that AidB is a marker of new poles and constriction sites, that could be considered as sites of preparation of new poles in the sibling cells originating from cell division. The possible role of AidB in the generation or the function of new poles needs further investigation.

  9. Chlamydiaceae Genomics Reveals Interspecies Admixture and the Recent Evolution of Chlamydia abortus Infecting Lower Mammalian Species and Humans.

    Science.gov (United States)

    Joseph, Sandeep J; Marti, Hanna; Didelot, Xavier; Castillo-Ramirez, Santiago; Read, Timothy D; Dean, Deborah

    2015-10-27

    Chlamydiaceae are obligate intracellular bacteria that cause a diversity of severe infections among humans and livestock on a global scale. Identification of new species since 1989 and emergence of zoonotic infections, including abortion in women, underscore the need for genome sequencing of multiple strains of each species to advance our knowledge of evolutionary dynamics across Chlamydiaceae. Here, we genome sequenced isolates from avian, lower mammalian and human hosts. Based on core gene phylogeny, five isolates previously classified as Chlamydia abortus were identified as members of Chlamydia psittaci and Chlamydia pecorum. Chlamydia abortus is the most recently emerged species and is a highly monomorphic group that lacks the conserved virulence-associated plasmid. Low-level recombination and evidence for adaptation to the placenta echo evolutionary processes seen in recently emerged, highly virulent niche-restricted pathogens, such as Bacillus anthracis. In contrast, gene flow occurred within C. psittaci and other Chlamydiaceae species. The C. psittaci strain RTH, isolated from a red-tailed hawk (Buteo jamaicensis), is an outlying strain with admixture of C. abortus, C. psittaci, and its own population markers. An average nucleotide identity of less than 94% compared with other Chlamydiaceae species suggests that RTH belongs to a new species intermediary between C. psittaci and C. abortus. Hawks, as scavengers and predators, have extensive opportunities to acquire multiple species in their intestinal tract. This could facilitate transformation and homologous recombination with the potential for new species emergence. Our findings indicate that incubator hosts such as birds-of-prey likely promote Chlamydiaceae evolution resulting in novel pathogenic lineages.

  10. Detection of Brucella abortus DNA in aborted goats and sheep in Egypt by real-time PCR

    OpenAIRE

    Wareth, Gamal; Melzer, Falk; Tomaso, Herbert; Roesler, Uwe; Neubauer, Heinrich

    2015-01-01

    Background Brucellosis is a major zoonoses affects wide range of domesticated as well as wild animals. Despite the eradication program of brucellosis in Egypt, the disease is still endemic among cattle, buffaloes, sheep, goats, and camels. Results In the present study, abortion occurred naturally among 25 animals (10 cows, 5 buffaloes, 9 Egyptian Baladi goats and 1 ewe) shared the same pasture were investigated by real-time polymerase chain reaction (RT-PCR). DNA of Brucella (B.) abortus was ...

  11. Outer membrane vesicles from Brucella abortus promote bacterial internalization by human monocytes and modulate their innate immune response.

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    Cora N Pollak

    Full Text Available Outer membrane vesicles (OMVs released by some gram-negative bacteria have been shown to exert immunomodulatory effects that favor the establishment of the infection. The aim of the present study was to assess the interaction of OMVs from Brucella abortus with human epithelial cells (HeLa and monocytes (THP-1, and the potential immunomodulatory effects they may exert. Using confocal microscopy and flow cytometry, FITC-labeled OMVs were shown to be internalized by both cell types. Internalization was shown to be partially mediated by clathrin-mediated endocytosis. Pretreatment of THP-1 cells with Brucella OMVs inhibited some cytokine responses (TNF-α and IL-8 to E. coli LPS, Pam3Cys or flagellin (TLR4, TLR2 and TLR5 agonists, respectively. Similarly, pretreatment with Brucella OMVs inhibited the cytokine response of THP-1 cells to B. abortus infection. Treatment of THP-1 cells with OMVs during IFN-γ stimulation reduced significantly the inducing effect of this cytokine on MHC-II expression. OMVs induced a dose-dependent increase of ICAM-1 expression on THP-1 cells and an increased adhesion of these cells to human endothelial cells. The addition of OMVs to THP-1 cultures before the incubation with live B. abortus resulted in increased numbers of adhered and internalized bacteria as compared to cells not treated with OMVs. Overall, these results suggest that OMVs from B. abortus exert cellular effects that promote the internalization of these bacteria by human monocytes, but also downregulate the innate immune response of these cells to Brucella infection. These effects may favor the persistence of Brucella within host cells.

  12. Typing discrepancy between phenotypic and molecular characterization revealing an emerging biovar 9 variant of smooth phage-resistant B. abortus strain 8416 in China

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    YaoXia eKang

    2015-12-01

    Full Text Available A newly isolated smooth colony morphology phage-resistant (SPR strain 8416 isolated from a 45-year-old cattle farm cleaner with clinical features of brucellosis in China was reported. The most unusual phenotype was its resistance to two Brucella phages Tbilisi and Weybridge, but sensitive to Berkeley 2, a pattern similar to that of B. melitensis biovar 1. VITEK 2 biochemical identification system found that both strain 8416 and B. melitensis strains shared positive ILATk, but negative in other B. abortus strains. However, routine biochemical and phenotypic characteristics of strain 8416 were most similar to that of B. abortus biovar 9 except CO2 requirement. In addition, multiple PCR molecular typing assays including AMOS-PCR, B. abortus special PCR (B-ab PCR and a novel sub-biovar typing PCR, indicated that strain 8416 may belong to either biovar 3b or 9 of B. abortus. Surprisingly, further MLVA typing results showed that strain 8416 was most closely related to B. abortus biovar 3 in the Brucella MLVA database, primarily differing in 4 out of 16 screened loci. Therefore, due to the unusual discrepancy between phenotypic (biochemical reactions and particular phage lysis profile and molecular typing characteristics, strain 8416 couldn’t be exactly classified to any of the existing B. abortus biovars and might be a new variant of B. abortus biovar 9. The present study also indicates that the present phage typing scheme for Brucella spp. is subject to variation and the routine Brucella biovar typing needs further studies.

  13. Proteomics-based confirmation of protein expression and correction of annotation errors in the Brucella abortus genome

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    Tomaki Fadi

    2010-05-01

    Full Text Available Abstract Background 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. Results The proteomic data was compiled from several independent comparative studies of Brucella abortus that used either outer membrane blebs, cytosols, or whole bacteria grown in media, as well as intracellular bacteria recovered at different times following macrophage infection. We identified a total of 621 bacterial proteins that were differentially expressed in a condition-specific manner. For 305 of these proteins we provide the first experimental evidence of their expression. Using a custom-built protein sequence database, we uncovered 7 annotation errors. We provide experimental evidence of expression of 5 genes that were originally annotated as non-expressed pseudogenes, as well as start site annotation errors for 2 other genes. Conclusions An essential element for ensuring correct functional studies is the correspondence between reported genome sequences and subsequent proteomics studies. In this study, we have used proteomics evidence to confirm expression of multiple proteins previously considered to be putative, as well as correct annotation errors in the genome of Brucella abortus strain 2308.

  14. Nucleotide-Binding Oligomerization Domain-1 and -2 Play No Role in Controlling Brucella abortus Infection in Mice

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    Fernanda S. Oliveira

    2012-01-01

    Full Text Available Nucleotide-binding oligomerization domain proteins (NODs are modular cytoplasmic proteins implicated in the recognition of peptidoglycan-derived molecules. Further, several in vivo studies have demonstrated a role for Nod1 and Nod2 in host defense against bacterial pathogens. Here, we demonstrated that macrophages from NOD1-, NOD2-, and Rip2-deficient mice produced lower levels of TNF-α following infection with live Brucella abortus compared to wild-type mice. Similar reduction on cytokine synthesis was not observed for IL-12 and IL-6. However, NOD1, NOD2, and Rip2 knockout mice were no more susceptible to infection with virulent B. abortus than wild-type mice. Additionally, spleen cells from NOD1-, NOD2-, and Rip2-deficient mice showed unaltered production of IFN-γ compared to C57BL/6 mice. Taken together, this study demonstrates that NOD1, NOD2 and Rip2 are dispensable for the control of B. abortus during in vivo infection.

  15. Mutation of purD and purF genes further attenuates Brucella abortus strain RB51.

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    Truong, Quang Lam; Cho, Youngjae; Barate, Abhijit Kashinath; Kim, Suk; Watarai, Masahisa; Hahn, Tae-Wook

    2015-02-01

    In the present study, transposon mutagenesis was used to further attenuate Brucella abortus RB51 vaccine strain. Two purD and purF mutants were constructed, characterized and evaluated for attenuation via intracellular survival in murine macrophage-like RAW264.7 and HeLa cells, and by clearance in BALB/c mice. The purD and purF mutants showed significantly decreased intracellular survival, and complementation of these mutants with intact copies of purD or purF genes of RB51 strain was able to restore these defects. In addition, the pur mutants presented significantly lowered persistence in mice. Immunization with purD and purF mutants protected mice against a challenge with the virulent B. abortus strain 544 at a level similar to that of the parent RB51. These data suggest that genes encoding the early stages of purine biosynthesis (purD and purF) are required for intracellular survival and virulence of B. abortus.

  16. Reduced Susceptibility to Rifampicin and Resistance to Multiple Antimicrobial Agents among Brucella abortus Isolates from Cattle in Brazil.

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    Barbosa Pauletti, Rebeca; Reinato Stynen, Ana Paula; Pinto da Silva Mol, Juliana; Seles Dorneles, Elaine Maria; Alves, Telma Maria; de Sousa Moura Souto, Monalisa; Minharro, Silvia; Heinemann, Marcos Bryan; Lage, Andrey Pereira

    2015-01-01

    This study aimed to determine the susceptibility profile of Brazilian Brucella abortus isolates from cattle to eight antimicrobial agents that are recommended for the treatment of human brucellosis and to correlate the susceptibility patterns with origin, biotype and MLVA16-genotype of the strains. Screening of 147 B. abortus strains showed 100% sensitivity to doxycycline and ofloxacin, one (0.68%) strain resistant to ciprofloxacin, two strains (1.36%) resistant to streptomycin, two strains (1.36%) resistant to trimethoprim-sulfamethoxazole and five strains (3.40%) resistant to gentamicin. For rifampicin, three strains (2.04%) were resistant and 54 strains (36.73%) showed reduced sensitivity. Two strains were considered multidrug resistant. In conclusion, the majority of B. abortus strains isolated from cattle in Brazil were sensitive to the antimicrobials commonly used for the treatment of human brucellosis; however, a considerable proportion of strains showed reduced susceptibility to rifampicin and two strains were considered multidrug resistant. Moreover, there was no correlation among the drug susceptibility pattern, origin, biotype and MLVA16-genotypes of these strains.

  17. An influenza viral vector Brucella abortus vaccine induces good cross-protection against Brucella melitensis infection in pregnant heifers.

    Science.gov (United States)

    Tabynov, Kaissar; Ryskeldinova, Sholpan; Sansyzbay, Abylai

    2015-07-17

    Brucella melitensis can be transmitted and cause disease in cattle herds as a result of inadequate management of mixed livestock farms. Ideally, vaccines against Brucella abortus for cattle should also provide cross-protection against B. melitensis. Previously we created a novel influenza viral vector B. abortus (Flu-BA) vaccine expressing the Brucella ribosomal proteins L7/L12 or Omp16. This study demonstrated Flu-BA vaccine with adjuvant Montanide Gel01 provided 100% protection against abortion in vaccinated pregnant heifers and good cross-protection of the heifers and their calves or fetuses (90-100%) after challenge with B. melitensis 16M; the level of protection provided by Flu-BA was comparable to the commercial vaccine B. abortus S19. In terms of the index of infection and colonization of Brucella in tissues, both vaccines demonstrated significant (P=0.02 to P<0.0001) protection against B. melitensis 16M infection compared to the negative control group (PBS+Montanide Gel01). Thus, we conclude the Flu-BA vaccine provides cross-protection against B. melitensis infection in pregnant heifers.

  18. Immunoproteomic identification of immunodominant antigens independent of the time of infection in Brucella abortus 2308-challenged cattle.

    Science.gov (United States)

    Lee, Jin Ju; Simborio, Hannah Leah; Reyes, Alisha Wehdnesday Bernardo; Kim, Dae Geun; Hop, Huynh Tan; Min, Wongi; Her, Moon; Jung, Suk Chan; Yoo, Han Sang; Kim, Suk

    2015-03-01

    Brucellosis is a vital zoonotic disease caused by Brucella, which infects a wide range of animals and humans. Accurate diagnosis and reliable vaccination can control brucellosis in domestic animals. This study examined novel immunogenic proteins that can be used to detect Brucella abortus infection or as an effective subcellular vaccine. In an immunoproteomic assay, 55 immunodominant proteins from B. abortus 544 were observed using two dimensional electrophoresis (2DE) and immunoblot profiles with antisera from B. abortus-infected cattle at the early (week 3), middle (week 7), and late (week 10) periods, after excluding protein spots reacting with antisera from Yersinia enterocolitica O:9-infected and non-infected cattle. Twenty-three selected immunodominant proteins whose spots were observed at all three infection periods were identified using MALDI-MS/MS. Most of these proteins identified by immunoblot and mass spectrometry were determined by their subcellular localization and predicted function. We suggest that the detection of prominent immunogenic proteins during the infection period can support the development of advanced diagnostic methods with high specificity and accuracy; subsidiarily, these proteins can provide supporting data to aid in developing novel vaccine candidates.

  19. Immunotoxic effect of thiamethoxam in immunized mice with Brucella abortus cultural filtrate antigen

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    L. H. Salema

    2016-12-01

    Full Text Available Aim: This study was planned for determination the toxic effect of thiamethoxam (TMX in immunized mice with Brucella abortus culture filtrate antigen (CFBAgs (as a vaccine and its role of TMX on decrease activity of B. abortus antigen on eliciting of humoral and cellular immunity. Materials and Methods: To achieve these goals 60 female mice were used, 7-8 weeks age, they were divided equally into three groups (20 in each group and treated as follows: 1st group: Mice were immunized with CFBAgs intraperitoneally in two doses, 2 weeks intervals with (protein concentration 2 mg\\ml, 2nd group: Mice immunized as in the 1st group and was administrated orally with 1/10 lethal dose 50% of TMX (83.7 mg/kg B.W. for 4 weeks daily, 3rd group was administrated orally with 0.3 ml normal saline served as a control group. At day 28 post immunization (PI delayed type hypersensitivity (skin test was done, and serum samples were collected at day 30 (PI for detection of passive hemagglutination test (PHA; interferon gamma (IFN-γ which was done by enzyme-linked immunosorbent assay test in addition to phagocytes assay. Results: The results of skin test post injection with soluble antigen of B. abortus intradermally showed a high significantly mean values at p≤0.05 of footpad skin thickness in the 1st group of mice which recorded (0.51±0.002 mm as compared with the 2nd group of mice which showed (0.08±0.002 mm after 24 h; the mean values of skin thickness were declined in the 1st mice (0.46±0.002 and 2nd mice (0.070±0.001 at 48 h; control group showed a negative results. These results were agreed with results of serum levels of IFN-γ (pg/ml that showed that a significant increase the vaccinated 1st group (406.36±1.52, than those values in the 2nd group (151.61±0.89 and negative result in 3rd group (46.47±0.60, in addition to results of PHA test which showed a significant increase in antibody titer in the 1st group (139±12.16 with low level of serum antibody

  20. Immunotoxic effect of thiamethoxam in immunized mice with Brucella abortus cultural filtrate antigen

    Science.gov (United States)

    Salema, L. H.; Alwan, M. J.; Yousif, Afaf Abdulrahman

    2016-01-01

    Aim: This study was planned for determination the toxic effect of thiamethoxam (TMX) in immunized mice with Brucella abortus culture filtrate antigen (CFBAgs) (as a vaccine) and its role of TMX on decrease activity of B. abortus antigen on eliciting of humoral and cellular immunity. Materials and Methods: To achieve these goals 60 female mice were used, 7-8 weeks age, they were divided equally into three groups (20 in each group) and treated as follows: 1st group: Mice were immunized with CFBAgs intraperitoneally in two doses, 2 weeks intervals with (protein concentration 2 mg\\ml), 2nd group: Mice immunized as in the 1st group and was administrated orally with 1/10 lethal dose 50% of TMX (83.7 mg/kg B.W.) for 4 weeks daily, 3rd group was administrated orally with 0.3 ml normal saline served as a control group. At day 28 post immunization (PI) delayed type hypersensitivity (skin test) was done, and serum samples were collected at day 30 (PI) for detection of passive hemagglutination test (PHA); interferon gamma (IFN-γ) which was done by enzyme-linked immunosorbent assay test in addition to phagocytes assay. Results: The results of skin test post injection with soluble antigen of B. abortus intradermally showed a high significantly mean values at p≤0.05 of footpad skin thickness in the 1st group of mice which recorded (0.51±0.002 mm) as compared with the 2nd group of mice which showed (0.08±0.002 mm) after 24 h; the mean values of skin thickness were declined in the 1st mice (0.46±0.002) and 2nd mice (0.070±0.001) at 48 h; control group showed a negative results. These results were agreed with results of serum levels of IFN-γ (pg/ml) that showed that a significant increase the vaccinated 1st group (406.36±1.52), than those values in the 2nd group (151.61±0.89) and negative result in 3rd group (46.47±0.60), in addition to results of PHA test which showed a significant increase in antibody titer in the 1st group (139±12.16) with low level of serum antibody

  1. Survival of Brucella abortus aqpX mutant in fresh and ripened cheeses.

    Science.gov (United States)

    Santiago-Rodríguez, María Del Rosario; Díaz-Aparicio, Efrén; Arellano-Reynoso, Beatriz; García-Lobo, Juan M; Gimeno, Miquel; Palomares-Reséndiz, Erika G; Hernández-Castro, Rigoberto

    2015-02-01

    The objective of this work was to evaluate the survival of a Brucella abortus aqpX mutant during the elaboration and conservation of fresh and ripened cheeses at 4 °C and 24 °C. The pH values and water activity were monitored for each type of cheese. The fresh cheese was elaborated with raw milk inoculated with 6×10⁸ colony-forming units (CFU)/mL each of parental and mutant strain. Ripening cheeses were elaborated with both raw and pasteurized milk and inoculated with 12×10⁸ CFU/mL each of parental and mutant strains. In fresh cheese, survival was observed during elaboration and conservation for 7 days at 4 °C in mutant and parental strains. The number of survivors of the mutant strain was 10 times lower compared with the parental strain at pH 5 and a(w) of 0.930. In the cheese elaborated with raw milk and ripened at 24 °C, both strains survived until day 17 at pH 4.0 and a(w) of 0.89. However, when the cheese was elaborated with pasteurized milk, the parental strain survived until day 31 of ripening, and the mutant strain survived 24 days at pH 4 and a(w) of 0.886. The survival of the mutant strain showed a diminution of one logarithm during elaboration and ripening of cheese as compared with the parental strain. When the cheese was elaborated with raw milk and ripened at 4 °C, survival of the parental strain was 24 days, whereas the mutant strain survived only 17 days (pH 5 and a(w) 0.90). Regarding the cheese elaborated with pasteurized milk and maturated at 4 °C, both strains survived 31 days (pH 5 and a(w) 0.90), with the same survival diminution during elaboration and ripening. Our results show that in both types of cheese, the mutated aqpX strain survived 10 times less than the parental strain, which shows that the aqpX gene can be related to the survival of Brucella abortus in this type of cheese.

  2. Detection of Leptospira spp. and Brucella abortus antibodies in free-living jaguars (Panthera onca) in two protected areas of northern Pantanal, Brazil.

    Science.gov (United States)

    Onuma, Selma Samiko Miyazaki; Kantek, Daniel Luis Zanella; Crawshaw Júnior, Peter Gransden; Morato, Ronaldo Gonçalves; May-Júnior, Joares Adenilson; Morais, Zenaide Maria de; Ferreira Neto, José Soares; Aguiar, Daniel Moura de

    2015-01-01

    This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.

  3. DETECTION OF Leptospira spp. AND Brucella abortus ANTIBODIES IN FREE-LIVING JAGUARS (Panthera onca IN TWO PROTECTED AREAS OF NORTHERN PANTANAL, BRAZIL

    Directory of Open Access Journals (Sweden)

    Selma Samiko Miyazaki ONUMA

    2015-04-01

    Full Text Available This study aimed to assess the exposure of free-living jaguars (Panthera onca to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT. The Rose Bengal test was applied for B. abortus antibodies. Two (18.2% jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01. All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.

  4. Neutrophils Exert a Suppressive Effect on Th1 Responses to Intracellular Pathogen Brucella abortus

    Science.gov (United States)

    Ordoñez-Rueda, Diana; Arce-Gorvel, Vilma; Alfaro-Alarcón, Alejandro; Lepidi, Hubert; Malissen, Bernard; Malissen, Marie; Gorvel, Jean-Pierre; Moreno, Edgardo

    2013-01-01

    Polymorphonuclear neutrophils (PMNs) are the first line of defense against microbial pathogens. In addition to their role in innate immunity, PMNs may also regulate events related to adaptive immunity. To investigate the influence of PMNs in the immune response during chronic bacterial infections, we explored the course of brucellosis in antibody PMN-depleted C57BL/6 mice and in neutropenic mutant Genista mouse model. We demonstrate that at later times of infection, Brucella abortus is killed more efficiently in the absence of PMNs than in their presence. The higher bacterial removal was concomitant to the: i) comparatively reduced spleen swelling; ii) augmented infiltration of epithelioid histiocytes corresponding to macrophages/dendritic cells (DCs); iii) higher recruitment of monocytes and monocyte/DCs phenotype; iv) significant activation of B and T lymphocytes, and v) increased levels of INF-γ and negligible levels of IL4 indicating a balance of Th1 over Th2 response. These results reveal that PMNs have an unexpected influence in dampening the immune response against intracellular Brucella infection and strengthen the notion that PMNs actively participate in regulatory circuits shaping both innate and adaptive immunity. PMID:23458832

  5. Endocarditis por Brucella abortus: Reporte del primer caso en C.R

    Directory of Open Access Journals (Sweden)

    Manuel Antonio Villalobos-Zúñiga

    2011-09-01

    Full Text Available Paciente masculino de 36 años de edad, proveniente de la zona rural de Costa Rica, con un cuadro clínico de 8 meses de evolución de fiebre, mialgias, artralgias, pérdida de peso y lumbalgia; referido por la detección de un soplo de insuficiencia aórtica. El ecocardiograma reveló endocarditis de la válvula aórtica, y se obtuvieron 4 hemocultivos positivos por Brucella abortus biotipo 3, con serologías negativas por brucelosis. Se inició tratamiento con antibióticos y luego se le realizó un reemplazo valvular aórtico; 4 meses después ingresó con dolor torácico que se atribuyó a una oclusión de la arteria descendente anterior, demostrada angiográficamente, por posible embolismo. En la actualidad cursa clínicamente estable con manejo médico para su cardiopatía, sin recaída infecciosa.

  6. Improvement of the Brucella abortus B19 vaccine by its preparation in a glycerol based medium.

    Science.gov (United States)

    Sangari, F J; Agüero, J; García-Lobo, J M

    1996-03-01

    The Brucella abortus B19 vaccine strain differs from other Brucella strains in its sensitivity to erythritol. However, erythritol tolerant (Eri(t)) mutants arise from sensitive cultures of B19 at high rate, and may cause persistence and/or abortion when the vaccine is inoculated on adult cattle. Twelve different batches of B19 have been examined for the presence of Eri(t) mutants. All contained Eri(t) variants at a proportion ranging from 10(-4) to 10(-6). In order to eliminate these mutants from the vaccine cultures, we have developed a minimal medium with glycerol as the sole carbon source, named MMG30. Growth of the parental strain B19 (erythritol sensitive) in this medium was fairly good compared with the growth of its Eri(t) derivatives. Culture of the 12 different batches of B19 in liquid MMG30 produced up to a thousandfold decrease in the proportion of Eri(t) mutants present in the vaccine cultures. Use of this medium to grow B19 could represent an easy and considerable improvement of the vaccine, by the reduction of the presence of potentially dangerous Eri(t) mutants.

  7. Immune Modulation of Recombinant OmpA against Brucella abortus 544 Infection in Mice.

    Science.gov (United States)

    Simborio, Hannah Leah Tadeja; Reyes, Alisha Wehdnesday Bernardo; Hop, Huynh Tan; Arayan, Lauren Togonon; Min, Wongi; Lee, Hu Jang; Lee, Jin Ju; Chang, Hong Hee; Kim, Suk

    2016-03-01

    Brucellosis affects a wide range of host species, including humans and many livestock animals. Chronic infections of the disease make antibiotic treatment costly, and the current vaccine used in livestock has not been approved for human use. This study investigated the possible use of the Brucella abortus outer membrane protein A (OmpA) as a candidate subunit vaccine in an infected mouse model. The ompA gene was cloned and overexpressed, and the recombinant OmpA (rOmpA) protein fused to maltose binding protein (MBP) was purified in Escherichia coli. Immunogenicity was verified through western blotting, and mice were immunized and challenged to evaluate its protective effect. Mice treated with rOmpA exhibited induced humoral and host cell-mediated responses, with a significant increase in immunoglobulin G (IgG1 and IgG2a) and cytokine levels, especially TNF-α and IL-12, compared with the control groups treated with either MBP or PBS. In conclusion, rOmpA should be highly considered as a future subunit vaccine for brucellosis, and further studies regarding rOmpA and its protective ability are suggested.

  8. Brucella abortus choloylglycine hydrolase affects cell envelope composition and host cell internalization.

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    María Inés Marchesini

    Full Text Available Choloylglycine hydrolase (CGH, E.C. 3.5.1.24 is a conjugated bile salt hydrolase that catalyses the hydrolysis of the amide bond in conjugated bile acids. Bile salt hydrolases are expressed by gastrointestinal bacteria, and they presumably decrease the toxicity of host's conjugated bile salts. Brucella species are the causative agents of brucellosis, a disease affecting livestock and humans. CGH confers Brucella the ability to deconjugate and resist the antimicrobial action of bile salts, contributing to the establishment of a successful infection through the oral route in mice. Additionally, cgh-deletion mutant was also attenuated in intraperitoneally inoculated mice, which suggests that CGH may play a role during systemic infection other than hydrolyzing conjugated bile acids. To understand the role CGH plays in B. abortus virulence, we infected phagocytic and epithelial cells with a cgh-deletion mutant (Δcgh and found that it is defective in the internalization process. This defect along with the increased resistance of Δcgh to the antimicrobial action of polymyxin B, prompted an analysis of the cell envelope of this mutant. Two-dimensional electrophoretic profiles of Δcgh cell envelope-associated proteins showed an altered expression of Omp2b and different members of the Omp25/31 family. These results were confirmed by Western blot analysis with monoclonal antibodies. Altogether, the results indicate that Brucella CGH not only participates in deconjugation of bile salts but also affects overall membrane composition and host cell internalization.

  9. Neutrophils exert a suppressive effect on Th1 responses to intracellular pathogen Brucella abortus.

    Directory of Open Access Journals (Sweden)

    Elías Barquero-Calvo

    2013-02-01

    Full Text Available Polymorphonuclear neutrophils (PMNs are the first line of defense against microbial pathogens. In addition to their role in innate immunity, PMNs may also regulate events related to adaptive immunity. To investigate the influence of PMNs in the immune response during chronic bacterial infections, we explored the course of brucellosis in antibody PMN-depleted C57BL/6 mice and in neutropenic mutant Genista mouse model. We demonstrate that at later times of infection, Brucella abortus is killed more efficiently in the absence of PMNs than in their presence. The higher bacterial removal was concomitant to the: i comparatively reduced spleen swelling; ii augmented infiltration of epithelioid histiocytes corresponding to macrophages/dendritic cells (DCs; iii higher recruitment of monocytes and monocyte/DCs phenotype; iv significant activation of B and T lymphocytes, and v increased levels of INF-γ and negligible levels of IL4 indicating a balance of Th1 over Th2 response. These results reveal that PMNs have an unexpected influence in dampening the immune response against intracellular Brucella infection and strengthen the notion that PMNs actively participate in regulatory circuits shaping both innate and adaptive immunity.

  10. Improving the molecular diagnosis of Chlamydia psittaci and Chlamydia abortus infection with a species-specific duplex real-time PCR.

    Science.gov (United States)

    Opota, Onya; Jaton, Katia; Branley, James; Vanrompay, Daisy; Erard, Veronique; Borel, Nicole; Longbottom, David; Greub, Gilbert

    2015-10-01

    Chlamydia psittaci and Chlamydia abortus are closely related intracellular bacteria exhibiting different tissue tropism that may cause severe but distinct infection in humans. C. psittaci causes psittacosis, a respiratory zoonotic infection transmitted by birds. C. abortus is an abortigenic agent in small ruminants, which can also colonize the human placenta and lead to foetal death and miscarriage. Infections caused by C. psittaci and C. abortus are underestimated mainly due to diagnosis difficulties resulting from their strict intracellular growth. We developed a duplex real-time PCR to detect and distinguish these two bacteria in clinical samples. The first PCR (PCR1) targeted a sequence of the 16S-23S rRNA operon allowing the detection of both C. psittaci and C. abortus. The second PCR (PCR2) targeted the coding DNA sequence CPSIT_0607 unique to C. psittaci. The two PCRs showed 100 % detection for ≥ 10 DNA copies per reaction (1000 copies ml(- 1)). Using a set of 120 samples, including bacterial reference strains, clinical specimens and infected cell culture material, we monitored 100 % sensitivity and 100 % specificity for the detection of C. psittaci and C. abortus for PCR1. When PCR1 was positive, PCR2 could discriminate C. psittaci from C. abortus with a positive predictive value of 100 % and a negative predictive value of 88 %. In conclusion, this new duplex PCR represents a low-cost and time-saving method with high-throughput potential, expected to improve the routine diagnosis of psittacosis and pregnancy complication in large-scale screening programs and also during outbreaks.

  11. Comparative evaluation of the protective efficacy of two formulations of a recombinant Chlamydia abortus subunit candidate vaccine in a mouse model.

    Science.gov (United States)

    Pan, Qing; Pais, Roshan; Ohandjo, Adaugo; He, Cheng; He, Qing; Omosun, Yusuf; Igietseme, J U; Eko, F O

    2015-04-08

    Chlamydia abortus (C. abortus) is the causative agent of ovine enzootic abortion (OEA) and poses a zoonotic risk to pregnant women. Current live attenuated 1B vaccines are efficacious but cause disease in vaccinated animals and inactivated vaccines are only marginally protective. We tested the ability of a new C. abortus subunit vaccine candidate based on the conserved and immunogenic polymorphic membrane protein D (Pmp18D) formulated in CpG1826+FL (Fms-like tyrosine kinase 3 Ligand; Flt3L) or Vibrio cholerae ghosts (VCG) to induce innate and cross protective immunity against genital C. abortus infection. We found that delivery of rPmp18D with VCG was more effective than with CpG+FL in up-regulating the expression of molecules critically involved in T cell activation and differentiation, including MHC II, CD40, CD80, and CD86, activation of TLRs and NLRP3 inflammasome engagement, and secretion of IL-1β and TNF-α but not IL-10 and IL-4. rVCG-Pmp18D-immunized mice elicited more robust antigen-specific IFN-γ, IgA and IgG2c antibody responses compared to CpG+FL-delivered rPmp18D. Based on the number of mice with positive vaginal cultures, length of vaginal shedding, and number of inclusion forming units recovered following challenge with the heterologous C. abortus strain B577, vaccine delivery with VCG induced superior protective immunity than delivery with a combination of CpG1826 and FL, a nasal DC-targeting adjuvant. These results demonstrate that the ability of VCG to enhance protective immunity against genital C. abortus infection is superior to that of CpG+FL adjuvants.

  12. Comparison of Buffered, Acidified Plate Antigen to Standard Serologic Tests for the Detection of Serum Antibodies to Brucella abortus in Elk (Cervus canadensis).

    Science.gov (United States)

    Clarke, P Ryan; Edwards, William H; Hennager, Steven G; Block, Jean F; Yates, Angela M; Ebel, Eric; Knopp, Douglas J; Fuentes-Sanchez, Antonio; Jennings-Gaines, Jessica; Kientz, Rebecca L; Simunich, Marilyn

    2015-07-01

    Brucellosis (caused by the bacterium Brucella abortus) is a zoonotic disease endemic in wild elk (Cervus canadensis) of the Greater Yellowstone Ecosystem, US. Because livestock and humans working with elk or livestock are at risk, validated tests to detect the B. abortus antibody in elk are needed. Using the κ-statistic, we evaluated the buffered, acidified plate antigen (BAPA) assay for agreement with the results of the four serologic tests (card test [card], complement fixation test [CF], rivanol precipitation plate agglutination test [RIV], standard plate agglutination test [SPT]) that are approved by the US Department of Agriculture for the detection of the B. abortus antibody in elk. From 2006 to 2010, serum samples collected from elk within B. abortus-endemic areas (n = 604) and nonendemic areas (n = 707) and from elk culture-positive for B. abortus (n = 36) were split and blind tested by four elk serum diagnostic laboratories. κ-Values showed a high degree of agreement for the card (0.876), RIV (0.84), and CF (0.774) test pairings and moderate agreement for the SPT (0.578). Sensitivities for the BAPA, card, RIV, CF, and SPT were 0.859, 0.839, 0.899, 1.00, and 0.813, whereas specificities were 0.986, 0.993, 0.986, 0.98, and 0.968, respectively. The positive predictive values and the negative predictive values were calculated for 2.6%, 8.8%, and 16.2% prevalence levels. These findings suggest the BAPA test is a suitable screening test for the B. abortus antibodies in elk.

  13. Estudio bacteriológico y serológico de brucelosis en vacas revacunadas con dosis reducida de cepa 19 de brucella abortus

    OpenAIRE

    Jorge Bustamante Sánchez; Fernando Israel Salazar Hernández; Efrén Díaz Aparicio; Carlos Manzano Cañas; Rafael Pérez González; Laura Hernández Andrade

    2000-01-01

    El objetivo fue determinar mediante pruebas diagnósticas, si los anticuerpos que se presentan en bovinos revacunados con dosis reducida de cepa 19 de Brucella abortus son debidos a infección o a vacunación, e intentar el aislamiento para determinar su especie y biotipo. El muestreo se realizó en 25 establos de Tizayuca, Hidalgo. Se utilizaron 100 vacas Holstein positivas a la prueba de tarjeta (PT), que habían sido vacunadas de tres a seis meses de edad con cepa 19 de B. abortus a dosis clási...

  14. ANTI-Lentivirus, Brucella abortus AND B. ovis ANTIBODIES IN SMALL RUMINANTS RAISED IN PERNAMBUCO AND BAHIA

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    RODOLFO DE MORAES PEIXOTO

    2016-01-01

    Full Text Available Goat and sheep production in the semi-arid northeast of Brazil has shown great economic potential. However, health problems can compromise the productivity of these animals. Given the scarcity of studies about the occurrence of these diseases, the aim of the present study was to analyze the serological diagnosis of anti-Brucella and anti-lentivirus antibodies among small ruminants in municipalities located in the Brazilian states of Bahia and Pernambuco. The samples were collected from local slaughterhouses and dairy farms. In total, 997 serum samples from animals in slaughterhouses and dairy herds were collected. In order to diagnose the caprine arthritis encephalitis virus (CAEV, the samples underwent agarose gel immunodiffusion (AGID testing. The buffered acidified antigen test (goats and agarose gel immunodiffusion test (sheep were used to detect anti-Brucella abortus and B. ovis antibodies following the methodology recommended by the Institute of Technology of Paraná (TECPAR. With anti-CAEV antibodies, seropositivity rates of 4.1% and 2.2% were recorded for animals from the slaughterhouses and dairy farms, respectively. None of the animals (goats or sheep were positive for anti-B. abortus antibodies. With B. ovis, a seropositivity rate of 6.5% (n = 13 was recorded among the 199 sheep serum samples. Results of the present study confirmed the presence of the CAE virus in the meat and dairy herds studied, although the prevalence was low. Natural infection by B. abortus did not occur in the goat and sheep herds assessed. Seropositivity for B. ovis was confirmed, although prevalence was low. Direct tests are required to diagnose ovine brucellosis.

  15. Overexpression of Brucella putative glycosyltransferase WbkA in B. abortus RB51 leads to production of exopolysaccharide

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    Neha eDabral

    2015-06-01

    Full Text Available Brucella spp. are Gram-negative, facultative intracellular bacteria that cause brucellosis in mammals. Brucella strains containing the O-polysaccharide in their cell wall structure exhibit a smooth phenotype whereas the strains devoid of the polysaccharide show rough phenotype. B. abortus strain RB51 is a stable rough attenuated mutant which is used as a licensed live vaccine for bovine brucellosis. Previous studies have shown that the wboA gene, which encodes a glycosyltransferase required for the synthesis of O-polysaccharide, is disrupted in B. abortus RB51 by an IS711 element. Although complementation of strain RB51 with a functional wboA gene results in O-polysaccharide synthesis in the cytoplasm, it does not result in smooth phenotype. The aim of this study was to determine if overexpression of Brucella WbkA or WbkE, two additional putative glycosyltransferases essential for O-polysaccharide synthesis, in strain RB51 would result in the O-polysaccharide synthesis and smooth phenotype. Our results demonstrate that overexpression of wbkA or wbkE gene in RB51 does not result in O-polysaccharide expression as shown by Western blotting with specific antibodies. However, wbkA, but not wbkE, overexpression leads to the development of a clumping phenotype and the production of exopolysaccharide(s containing mannose, galactose, N-acetylglucosamine and N-acetylgalactosamine. Moreover, we found that the clumping recombinant strain displays increased adhesion to polystyrene plates. The recombinant strain was similar to strain RB51 in its attenuation characteristic and in its ability to induce protective immunity against virulent B. abortus challenge in mice.

  16. Survey of Omp19 immunogenicity against Brucella abortus and Brucella melitensis: influence of nanoparticulation versus traditional immunization.

    Science.gov (United States)

    Abkar, Morteza; Lotfi, Abbas Sahebghadam; Amani, Jafar; Eskandari, Khadijeh; Ramandi, Mehdi Fasihi; Salimian, Jafar; Brujeni, Gholamreza Nikbakht; Alamian, Saeed; Kamali, Mehdi; Koushki, Hamid

    2015-12-01

    Brucellosis is the most common zoonotic bacterial disease. Prevention of human brucellosis is achieved through pasteurization of dairy products, appropriate sanitation and vaccination of domestic animals against the Brucella species. B. abortus unlipidated 19 kDa outer membrane protein (U-Omp19) is a promising candidate for a subunit vaccine against brucellosis. This study investigates immunogenicity of Omp19 alone and with Freund's adjuvant (Omp19-IFA) and N-trimethyl chitosan (TMC/Omp19) nanoparticles, as well as the effect of Omp19 administration route on immunological responses and protection. The omp19 gene was expressed in E. coli BL21 (DE3). After purification, the recombinant Omp19 was loaded onto TMC nanoparticles by ionic gelation with tripolyphosphate. Particle size and loading efficiency of the nanoparticles were determined. Omp19-IFA was administered intraperitoneally while TMC/Omp19 nanoparticles were administered orally and intraperitoneally. The results indicated that intraperitoneal (i.p.) immunization by Omp19-IFA and TMC/Omp19 nanoparticles induced Th1 and Th2 immune responses, respectively, whereas oral immunization of TMC/Omp19 nanoparticles induced a mixed Th1/Th17 immune response. Moreover, oral immunization increased IgA levels in feces. Immunized mice were challenged with virulent B. melitensis 16 M and B. abortus 544. Oral immunization with TMC/Omp19 nanoparticles induced a remarkably high protection level against B. melitensis and B. abortus. The results showed that immunization route has a pivotal role in immune response polarization and protective efficiency of Omp19 antigen. Also, it was deduced that the higher protection level achieved through oral administration of TMC/Omp19 nanoparticles may be due to the elicited Th17 response.

  17. Overexpression of Brucella putative glycosyltransferase WbkA in B. abortus RB51 leads to production of exopolysaccharide.

    Science.gov (United States)

    Dabral, Neha; Jain-Gupta, Neeta; Seleem, Mohamed N; Sriranganathan, Nammalwar; Vemulapalli, Ramesh

    2015-01-01

    Brucella spp. are Gram-negative, facultative intracellular bacteria that cause brucellosis in mammals. Brucella strains containing the O-polysaccharide in their cell wall structure exhibit a smooth phenotype whereas the strains devoid of the polysaccharide show rough phenotype. B. abortus strain RB51 is a stable rough attenuated mutant which is used as a licensed live vaccine for bovine brucellosis. Previous studies have shown that the wboA gene, which encodes a glycosyltransferase required for the synthesis of O-polysaccharide, is disrupted in B. abortus RB51 by an IS711 element. Although complementation of strain RB51 with a functional wboA gene results in O-polysaccharide synthesis in the cytoplasm, it does not result in smooth phenotype. The aim of this study was to determine if overexpression of Brucella WbkA or WbkE, two additional putative glycosyltransferases essential for O-polysaccharide synthesis, in strain RB51 would result in the O-polysaccharide synthesis and smooth phenotype. Our results demonstrate that overexpression of wbkA or wbkE gene in RB51 does not result in O-polysaccharide expression as shown by Western blotting with specific antibodies. However, wbkA, but not wbkE, overexpression leads to the development of a clumping phenotype and the production of exopolysaccharide(s) containing mannose, galactose, N-acetylglucosamine, and N-acetylgalactosamine. Moreover, we found that the clumping recombinant strain displays increased adhesion to polystyrene plates. The recombinant strain was similar to strain RB51 in its attenuation characteristic and in its ability to induce protective immunity against virulent B. abortus challenge in mice.

  18. DNA vaccine encoding L7/L12-P39 of Brucella abortus induces protective immunity in BALB/c mice

    Institute of Scientific and Technical Information of China (English)

    LUO De-yan; LI Peng; XING Li; ZHAO Guang-yu; SHI Wei; ZHANG Song-le; WANG Xi-liang

    2006-01-01

    @@ Brucella abortus is a gram-negative, facultative, intracellular bacterium that infects both cattle and humans, causing abortion and infertility in the former and undulant fever, endocarditis, arthritis, and osteomyelitis. Resistance to Brucella depends on acquired cell-mediated immunity (CMI).1 Live attenuated vaccines can stimulate strong CMI response, which are usually very effective against brucellosis and are used to control brucellosis in domestic animals. However, there is no safe and effective vaccine available for human because the vaccine strains used for animals are considered too virulent for humans. A vaccine that will be noninfectious to humans but effective in stimulating a broad protective immune response is needed.2

  19. Detection of Brucella abortus by alkB and IS711 based primers

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    Seyed Reza Hosseini Doust

    2007-06-01

    Full Text Available

    BACKGROUND: Brucellosis is a zoonotic disease, which involves both animals and human. Although the conventional methods have been widely used for its laboratory diagnosis, the PCR techniques have proved to be useful due to specificity, sensitivity and the rapidness. Various target sequences of brucella bacterium such as OMP2, 16s RNA and IS711 have been used for the primer designing.. All primer sets have shown different sensitivities and specificities. In present investigation, PCR protocol and primer designated based on IS711 and a fragment of chromosomal DNA all were optimized with standard genome and clinical samples.

    METHODS: Numerous tissue samples (liver, kidney, lymph node, and uterus were prepared and were cultured by the bacteriological standard methods along with the serology positive human samples. PCR protocol was optimized and the primer's sensitivity and the specificity were checked using pure genome of B. abortus. All samples were tested by the standard bacteriological methods. The samples were then subject to PCR amplification and the PCR product was confirmed using the RFLP technique.

    RESULTS: The culture results indicated a poor sensitivity as it was previously reported. The PCR product 157 bp was observed on the agarose gel indicating that significant number of clinical samples (human brucellosis cases were positive by PCR but not by the

  20. Isolation of two biologically active cell surface proteins from Brucella abortus by chromatofocusing

    Energy Technology Data Exchange (ETDEWEB)

    Tabatabai, L.B.; Deyoe, B.L.

    1983-01-01

    Brucella abortus contains a group of immunogenic cell surface proteins which have potential value as a vaccine or as a diagnostic reagent for the prevention and diagnosis of bovine brucellosis. Under nondenaturing conditions, these proteins range in molecular weight from 10,000-124,000, as determined by high performance liquid chromatography (HPLC) on TSK 3000sw. By analytical isoelectrofocusing, 6 major protein bands could be distinguished with pI's ranging from 4.0 to 6.0 and 3 additional major proteins with pI's of 7.5, 9.5, and 10. By chromatofocusing on Polybuffer Exchanger 94 with a pH gradient from 6-4, two of the six proteins from pI 4-6 were separated, a pI 4.9 and a pI 4.7 protein; a third fraction contained the high pI proteins. The former two proteins were homogeneous by analytical isoelectrofocusing, and a molecular weight of 54,000 daltons was found for both protein species by HPLC on TSK 3000sw. The pI 4-6 and not the pI 9.5 and 10 proteins, could be radiolabeled when intact cells were radioiodinated with diazotized (/sup 125/I)-iodosulfanilic acid. Biological activity of the proteins as assessed in lemmings indicated that immunization with the pI 4.7 and 4.9 proteins afforded better protection against experimental brucellosis than immunization with the high pI proteins. These results support our view that a single surface protein may be sufficient for the prevention of experimental brucellosis.

  1. Prediction of T cell epitopes of Brucella abortus and evaluation of their protective role in mice.

    Science.gov (United States)

    Afley, Prachiti; Dohre, Sudhir K; Prasad, G B K S; Kumar, Subodh

    2015-09-01

    Brucellae are Gram-negative intracellular bacteria that cause an important zoonotic disease called brucellosis. The animal vaccines are available but have disadvantage of causing abortions in a proportion of pregnant animals. The animal vaccines are also pathogenic to humans. Recent trend in vaccine design has shifted to epitope-based vaccines that are safe and specific. In this study, efforts were made to identify MHC-I- and MHC-II-restricted T cell epitopes of Brucella abortus and evaluate their vaccine potential in mice. The peptides were designed using online available immunoinformatics tools, and five MHC-I- and one MHC-II-restricted T cell peptides were selected on the basis of their ability to produce interferon gamma (IFN-γ) in in vivo studies. The selected peptides were co-administered with poly DL-lactide-co-glycolide (PLG) microparticles and evaluated for immunogenicity and protection in BALB/c mice. Mice immunized with peptides either entrapped in PLG microparticles (EPLG-Pep) or adsorbed on PLG particles (APLG-Pep) showed significantly higher splenocyte proliferation and IFN-γ generation to all selected peptides than the mice immunized with corresponding irrelevant peptides formulated PLG microparticles or phosphate-buffered saline (PBS). A significant protection compared to PBS control was also observed in EPLG-Pep and APLG-Pep groups. A plasmid DNA vaccine construct (pVaxPep) for peptides encoding DNA sequences was generated and injected to mice by in vivo electroporation. Significant protection was observed (1.66 protection units) when compared with PBS and empty vector control group animals. Overall, the MHC-I and MHC-II peptides identified in this study are immunogenic and protective in mouse model and support the feasibility of peptide-based vaccine for brucellosis.

  2. The Assessment of Cytokine and Antibody Responses to Recombinant 31kDa Brucella Cell-Surface Protein in Brucella Abortus Infected Mouse Model

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    Nima Khoramabadi

    2014-06-01

    Conclusion: These findings suggest that specific humoral and cell-mediated responses to BCSP31 is formed during murine host infection with B. abortus. Based on these findings, rBCSP31 can be used in further design of immunogenic strategies for vaccination against brucellosis.

  3. The efficacy of the skin delayed-type hypersensitivity using a brucellin prepared from a mucoid strain of Brucella abortus to detect brucellosis

    NARCIS (Netherlands)

    Bercovich, Z.; Muskens, J.A.M.

    1998-01-01

    Eight-hundred-and-ninety-six cattle belonging to herds officially designated Brucella-free, and 190 cattle belonging to infected herds were tested with the skin delayed-type hypersensitivity (SDTH) test, using brucellin (273) prepared from a rnucoid strain of Brucella abortus. An increase in skinfol

  4. The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests

    NARCIS (Netherlands)

    Emmerzaal, A.; Wit, de J.J.; Dijkstra, T.; Bakker, D.; Ziiderveld, van F.G.

    2002-01-01

    The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the progra

  5. Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308.

    Science.gov (United States)

    Matrone, M; Keid, L B; Rocha, V C M; Vejarano, M P; Ikuta, C Y; Rodriguez, C A R; Ferreira, F; Dias, R A; Ferreira Neto, J S

    2009-07-01

    The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (pprotocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and vice-versa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol.

  6. Immune responses and protection against experimental challenge after vaccination of bison with Brucella abortus strains RB51 or RB51 overexpressing superoxide dismutase and Glycosyltransferase genes

    Science.gov (United States)

    Vaccination is a tool that could be beneficial in managing the high prevalence of brucellosis in free-ranging bison in Yellowstone National Park. In this study, we characterized immunologic responses and protection against experimental challenge after vaccination of bison with Brucella abortus stra...

  7. Phenotypic and genotypic characterization of Brucella strains isolated from autochthonous livestock reveals the dominance of B. abortus biovar 3a in Nigeria.

    Science.gov (United States)

    Bertu, Wilson J; Ducrotoy, Marie J; Muñoz, Pilar M; Mick, Virginie; Zúñiga-Ripa, Amaia; Bryssinckx, Ward; Kwaga, Jacob K P; Kabir, Junaid; Welburn, Susan C; Moriyón, Ignacio; Ocholi, Reuben A

    2015-10-22

    Brucellosis is a worldwide widespread zoonosis caused by bacteria of the genus Brucella. Control of this disease in a given area requires an understanding of the Brucella species circulating in livestock and humans. However, because of the difficulties intrinsic to Brucella isolation and typing, such data are scarce for resource-poor areas. The paucity of bacteriological data and the consequent imperfect epidemiological picture are particularly critical for Sahelian and Sub-Sahara African countries. Here, we report on the characterization of 34 isolates collected between 1976 and 2012 from cattle, sheep and horses in Nigeria. All isolates were identified as Brucella abortus by Bruce-ladder PCR and assigned to biovar 3 by conventional typing. Further analysis by enhanced AMOS-ERY PCR showed that all of them belonged to the 3a sub-biovar, and MLVA analysis grouped them in a cluster clearly distinct from that formed by European B. abortus biovar 3b strains. Nevertheless, MLVA detected heterogeneity within the Nigerian biovar 3a strains. The close genetic profiles of the isolates from cattle, sheep and horses, suggest that, at least in some parts of Nigeria, biovar 3a circulates among animal species that are not the preferential hosts of B. abortus. Consistent with previous genetic analyses of 7 strains from Ivory Cost, Gambia and Togo, the analysis of these 34 Nigerian strains supports the hypothesis that the B. abortus biovar 3a lineage is dominant in West African countries.

  8. Pseudorabies Virus and Brucella abortus from an Expanding Wild Pig ( Sus scrofa ) Population in Southern Oklahoma, USA.

    Science.gov (United States)

    Gaskamp, Joshua A; Gee, Kenneth L; Campbell, Tyler A; Silvy, Nova J; Webb, Stephen L

    2016-04-28

    Wild pigs ( Sus scrofa ) are causing increasing ecologic and economic damage at a global scale. Because wild pigs can carry ≥65 diseases that affect livestock, their widespread expansion threatens native wildlife and livestock. We screened wild pigs from south-central Oklahoma, US for antibodies against Brucella abortus , pseudorabies virus (PRV), and porcine reproductive and respiratory syndrome virus (PRRS). These pathogens were chosen because they are part of eradication programs in the US and could have large economic impacts on domestic livestock if transmitted from wild animals. We tested 282 serum samples during spring 2010 (n=149) and 2011 (n=133) and found an overall exposure rate to PRV of 24.1% (n=68); PRV was detected at two of three study sites. Two wild pigs had detectable antibody to B. abortus , and one had detectable antibody to PRRS. On average, 27% of wild pigs within a sounder were positive for PRV antibody, with 44% of the sounders (16/36) having at least one positive individual. These data highlight that wild pigs could carry pathogens that affect domestic livestock. Because the US is free of these pathogens in commercial livestock operations, continued surveillance and vaccination of domestic livestock are needed. Commercial livestock producers at the wildlife-livestock interface may benefit from spatial prioritization of risk zones to facilitate strategic control efforts.

  9. Glial Cell-Elicited Activation of Brain Microvasculature in Response to Brucella abortus Infection Requires ASC Inflammasome-Dependent IL-1β Production.

    Science.gov (United States)

    Miraglia, M Cruz; Costa Franco, Miriam M; Rodriguez, Ana M; Bellozi, Paula M Q; Ferrari, Carina C; Farias, Maria I; Dennis, Vida A; Barrionuevo, Paula; de Oliveira, Antonio C P; Pitossi, Fernando; Kim, Kwang Sik; Delpino, M Victoria; Oliveira, Sergio Costa; Giambartolomei, Guillermo H

    2016-05-01

    Blood-brain barrier activation and/or dysfunction are a common feature of human neurobrucellosis, but the underlying pathogenic mechanisms are largely unknown. In this article, we describe an immune mechanism for inflammatory activation of human brain microvascular endothelial cells (HBMEC) in response to infection with Brucella abortus Infection of HBMEC with B. abortus induced the secretion of IL-6, IL-8, and MCP-1, and the upregulation of CD54 (ICAM-1), consistent with a state of activation. Culture supernatants (CS) from glial cells (astrocytes and microglia) infected with B. abortus also induced activation of HBMEC, but to a greater extent. Although B. abortus-infected glial cells secreted IL-1β and TNF-α, activation of HBMEC was dependent on IL-1β because CS from B. abortus-infected astrocytes and microglia deficient in caspase-1 and apoptosis-associated speck-like protein containing a CARD failed to induce HBMEC activation. Consistently, treatment of CS with neutralizing anti-IL-1β inhibited HBMEC activation. Both absent in melanoma 2 and Nod-like receptor containing a pyrin domain 3 are partially required for caspase-1 activation and IL-1β secretion, suggesting that multiple apoptosis-associated speck-like protein containing CARD-dependent inflammasomes contribute to IL-1β-induced activation of the brain microvasculature. Inflammasome-mediated IL-1β secretion in glial cells depends on TLR2 and MyD88 adapter-like/TIRAP. Finally, neutrophil and monocyte migration across HBMEC monolayers was increased by CS from Brucella-infected glial cells in an IL-1β-dependent fashion, and the infiltration of neutrophils into the brain parenchyma upon intracranial injection of B. abortus was diminished in the absence of Nod-like receptor containing a pyrin domain 3 and absent in melanoma 2. Our results indicate that innate immunity of the CNS set in motion by B. abortus contributes to the activation of the blood-brain barrier in neurobrucellosis and IL-1β mediates

  10. Brucella abortus Inhibits Major Histocompatibility Complex Class II Expression and Antigen Processing through Interleukin-6 Secretion via Toll-Like Receptor 2▿

    Science.gov (United States)

    Barrionuevo, Paula; Cassataro, Juliana; Delpino, M. Victoria; Zwerdling, Astrid; Pasquevich, Karina A.; Samartino, Clara García; Wallach, Jorge C.; Fossati, Carlos A.; Giambartolomei, Guillermo H.

    2008-01-01

    The strategies that allow Brucella abortus to survive inside macrophages for prolonged periods and to avoid the immunological surveillance of major histocompatibility complex class II (MHC-II)-restricted gamma interferon (IFN-γ)-producing CD4+ T lymphocytes are poorly understood. We report here that infection of THP-1 cells with B. abortus inhibited expression of MHC-II molecules and antigen (Ag) processing. Heat-killed B. abortus (HKBA) also induced both these phenomena, indicating the independence of bacterial viability and involvement of a structural component of the bacterium. Accordingly, outer membrane protein 19 (Omp19), a prototypical B. abortus lipoprotein, inhibited both MHC-II expression and Ag processing to the same extent as HKBA. Moreover, a synthetic lipohexapeptide that mimics the structure of the protein lipid moiety also inhibited MHC-II expression, indicating that any Brucella lipoprotein could down-modulate MHC-II expression and Ag processing. Inhibition of MHC-II expression and Ag processing by either HKBA or lipidated Omp19 (L-Omp19) depended on Toll-like receptor 2 and was mediated by interleukin-6. HKBA or L-Omp19 also inhibited MHC-II expression and Ag processing of human monocytes. In addition, exposure to the synthetic lipohexapeptide inhibited Ag-specific T-cell proliferation and IFN-γ production of peripheral blood mononuclear cells from Brucella-infected patients. Together, these results indicate that there is a mechanism by which B. abortus may prevent recognition by T cells to evade host immunity and establish a chronic infection. PMID:17984211

  11. Serology for Brucella abortus in cart horses from an urban area in Brazil Sorologia para Brucella abortus em cavalos de carroça de área urbana do Brasil

    Directory of Open Access Journals (Sweden)

    J.M.A.P. Antunes

    2013-04-01

    Full Text Available Avaliou-se a infecção por Brucella abortus em cavalos de carroça de Curitiba e São José dos Pinhais-PR. Um total de 123 amostras foi submetido ao teste do antígeno tamponado acidificado (ATA, soroaglutinação lenta em tubos (SAL e prova do 2-mercaptoetanol (2-ME para confirmação dos resultados. Oito (6,5% equinos foram positivos para o ATA e um animal permaneceu positivo ao teste confirmatório. Existem evidências da presença de brucelose entre os cavalos de carroça.

  12. Diagnóstico sorológico da brucelose bovina em animais adultos vacinados com dose reduzida da cepa 19 de Brucella abortus Serological diagnosis of bovine brucellosis in adult herd vaccinated with Brucella abortus strain 19 reduced dose

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    Gustavo Coelho Jardim

    2006-09-01

    Full Text Available Com o presente trabalho avaliou-se o uso de dose reduzida da vacina produzida com a amostra 19 de Brucella abortus, em rebanho adulto negativo para a enfermidade, por meio de técnicas de diagnóstico sorológico preconizadas pelo Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Animal e por um ensaio indireto de imunoadsorção enzimática (ELISA ID. A prova de fixação de complemento detectou 46,77% de positivos, o antígeno acidificado tamponado 67,74%, o 2-mercaptoetanol com soroaglutinação lenta 87,09% e o ELISA ID 100%. A dose reduzida interferiu no diagnóstico sorológico. Nenhuma das técnicas apresentou especificidade adequada para uso em rebanho nestas condições, até 3 meses após a vacinação.The study evaluated the use of a reduced dose of the Brucella abortus strain 19 vaccine, in an adult herd negative for the disease, by serological diagnostic techniques, advocated by the Brazilian Program for Animal Brucellosis and Tuberculosis Control and Eradication, and by an indirect ELISA. The complement fixation test detecteed 46.77% positives, the rose bengal test 67.74%, the mercaptoethanol with standard agglutination test 87.09% and the ELISA ID 100%. The reduced dose influenced the serological diagnosis. None of the techniques reached a suitable specificity for use in the herd under those conditions, up to 3 months after vaccination.

  13. Seminal vesiculitis and orchitis caused by Brucella abortus biovar 1 in young bison bulls from South Dakota.

    Science.gov (United States)

    Rhyan, J C; Holland, S D; Gidlewski, T; Saari, D A; Jensen, A E; Ewalt, D R; Hennager, S G; Olsen, S C; Cheville, N F

    1997-10-01

    Specimens of blood, lymph nodes, spleens, and genitalia were collected at slaughter from seven 3- and 4-year-old male bison that had recently become seropositive for brucellosis. The animals were from a captive herd of approximately 3,500 bison located in central South Dakota. Brucella abortus biovar 1 was isolated from 2 or more specimens from each of 6 bison. Severe necrotizing and pyogranulomatous orchitis was present in 1 testicle from 1 bull, and 4 animals had mild to marked seminal vesiculitis. Immunohistochemical staining labeled organisms in seminal vesicles and the testicle with orchitis. Ultrastructurally, intact bacilli were present in cytoplasmic vacuoles of some macrophages; other macrophages contained intracytoplasmic aggregates of calcified coccobacilli.

  14. Viabilidade da Brucella abortus durante a cura de queijo parmesão fabricado com leite experimentalmente contaminado

    OpenAIRE

    2012-01-01

    A legislação brasileira permite o uso de leite cru na fabricação de queijos curados se o período de cura for superior a 60 dias (a 5°C ou mais). Entretanto, não há evidência científica sólida de que durante a cura ocorre suficiente inativação de Brucella abortus, sob a perspectiva da segurança dos alimentos. Além disso, não há metodologia oficial para quantificação de brucelas em matriz alimentar. Desta forma este projeto propõe um protocolo para estudos da curva de decaimento de Brucella abo...

  15. Crystallographic and kinetic study of riboflavin synthase from Brucella abortus, a chemotherapeutic target with an enhanced intrinsic flexibility

    Energy Technology Data Exchange (ETDEWEB)

    Serer, María I.; Bonomi, Hernán R. [IIBBA–CONICET, Avenida Patricias Argentinas 435, C1405BWE Buenos Aires (Argentina); Guimarães, Beatriz G. [Synchrotron SOLEIL, L’Orme des Merisiers, Saint-Aubin BP 48, 91192 Gif-sur-Yvette CEDEX (France); Rossi, Rolando C. [Universidad de Buenos Aires, Junín 956, C1113AAD Buenos Aires (Argentina); Goldbaum, Fernando A.; Klinke, Sebastián, E-mail: sklinke@leloir.org.ar [IIBBA–CONICET, Avenida Patricias Argentinas 435, C1405BWE Buenos Aires (Argentina)

    2014-05-01

    This work reports crystal structures of trimeric riboflavin synthase from the pathogen B. abortus both as the apo protein and in complex with several ligands of interest. It is shown that ligand binding drives the assembly of the unique active site of the trimer, and these findings are complemented by a detailed kinetic study on this enzyme, in which marked inhibition by substrate and product was observed. Riboflavin synthase (RS) catalyzes the last step of riboflavin biosynthesis in microorganisms and plants, which corresponds to the dismutation of two molecules of 6,7-dimethyl-8-ribityllumazine to yield one molecule of riboflavin and one molecule of 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione. Owing to the absence of this enzyme in animals and the fact that most pathogenic bacteria show a strict dependence on riboflavin biosynthesis, RS has been proposed as a potential target for antimicrobial drug development. Eubacterial, fungal and plant RSs assemble as homotrimers lacking C{sub 3} symmetry. Each monomer can bind two substrate molecules, yet there is only one active site for the whole enzyme, which is located at the interface between two neighbouring chains. This work reports the crystallographic structure of RS from the pathogenic bacterium Brucella abortus (the aetiological agent of the disease brucellosis) in its apo form, in complex with riboflavin and in complex with two different product analogues, being the first time that the structure of an intact RS trimer with bound ligands has been solved. These crystal models support the hypothesis of enhanced flexibility in the particle and also highlight the role of the ligands in assembling the unique active site. Kinetic and binding studies were also performed to complement these findings. The structural and biochemical information generated may be useful for the rational design of novel RS inhibitors with antimicrobial activity.

  16. Performance of skin tests with allergens from B. melitensis B115 and rough B. abortus mutants for diagnosing swine brucellosis.

    Science.gov (United States)

    Dieste-Pérez, L; Blasco, J M; De Miguel, M J; Marín, C M; Barberán, M; Conde-Álvarez, R; Moriyón, I; Muñoz, P M

    2014-01-10

    Swine brucellosis by Brucella suis biovar 2 is an emerging disease whose control is based on serological testing and culling. However, current serological tests detect antibodies to the O-polysaccharide (O/PS) moiety of Brucella smooth lipopolysaccharide (S-LPS), and thus lack specificity when infections by Yersinia enterocolitica O:9 and other gram-negative bacteria carrying cross-reacting O/PS occur. The skin test with the protein-rich brucellin extract obtained from rough B. melitensis B115 is assumed to be specific for discriminating these false positive serological reactions (FPSR). However, B115 strain, although unable to synthesize S-LPS, accumulates O/PS internally, which could cause diagnostic problems. Since the brucellin skin test has been seldom used in pigs and FPSR are common in these animals, we assessed its performance using cytosoluble protein extracts obtained from B. abortus rough mutants in manBcore or per genes (critical for O/PS biosynthesis) and B. melitensis B115. The diagnostic sensitivity and specificity were determined in B. suis biovar 2 culture positive and brucellosis free sows, and apparent prevalence in sows of unknown individual bacteriological and serological status belonging to B. suis biovar 2 naturally infected herds. Moreover, the specificity in discriminating brucellosis from FPSR was assessed in brucellosis free boars showing FPSR. The skin test with B. abortus ΔmanBcore and B. melitensis B115 allergens performed similarly, and the former one resulted in 100% specificity when testing animals showing FPSR in indirect ELISA, Rose Bengal and complement fixation serological tests. We conclude that O/PS-free genetically defined mutants represent an appropriate alternative to obtain Brucella protein extracts for diagnosing swine brucellosis.

  17. Brucella abortus uses a stealthy strategy to avoid activation of the innate immune system during the onset of infection.

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    Elías Barquero-Calvo

    Full Text Available BACKGROUND: To unravel the strategy by which Brucella abortus establishes chronic infections, we explored its early interaction with innate immunity. METHODOLOGY/PRINCIPAL FINDINGS: Brucella did not induce proinflammatory responses as demonstrated by the absence of leukocyte recruitment, humoral or cellular blood changes in mice. Brucella hampered neutrophil (PMN function and PMN depletion did not influence the course of infection. Brucella barely induced proinflammatory cytokines and consumed complement, and was strongly resistant to bactericidal peptides, PMN extracts and serum. Brucella LPS (BrLPS, NH-polysaccharides, cyclic glucans, outer membrane fragments or disrupted bacterial cells displayed low biological activity in mice and cells. The lack of proinflammatory responses was not due to conspicuous inhibitory mechanisms mediated by the invading Brucella or its products. When activated 24 h post-infection macrophages did not kill Brucella, indicating that the replication niche was not fusiogenic with lysosomes. Brucella intracellular replication did not interrupt the cell cycle or caused cytotoxicity in WT, TLR4 and TLR2 knockout cells. TNF-alpha-induction was TLR4- and TLR2-dependent for live but not for killed B. abortus. However, intracellular replication in TLR4, TLR2 and TLR4/2 knockout cells was not altered and the infection course and anti-Brucella immunity development upon BrLPS injection was unaffected in TLR4 mutant mice. CONCLUSION/SIGNIFICANCE: We propose that Brucella has developed a stealth strategy through PAMPs reduction, modification and hiding, ensuring by this manner low stimulatory activity and toxicity for cells. This strategy allows Brucella to reach its replication niche before activation of antimicrobial mechanisms by adaptive immunity. This model is consistent with clinical profiles observed in humans and natural hosts at the onset of infection and could be valid for those intracellular pathogens phylogenetically

  18. Isolation of a field strain of Brucella abortus from RB51-vaccinated- and brucellosis-seronegative bovine yearlings that calved normally.

    Science.gov (United States)

    Arellano-Reynoso, Beatriz; Suárez-Güemes, Francisco; Estrada, Félix Mejía; Michel-GómezFlores, Fernando; Hernández-Castro, Rigoberto; Acosta, Rómulo Beltrán; Díaz-Aparicio, Efrén

    2013-02-01

    A study was carried out in Pichucalco, Chiapas (Mexico) to determine whether recently calved cows or those that aborted shed Brucella. Serological diagnosis of brucellosis was made in all animals (209). Six of the cows that calved normally and two that aborted underwent a bacteriological study of milk and vaginal exudate. Brucella abortus was isolated from vaginal exudate samples in two 3- to 4-year-old seronegative first-birth cows that had calved normally. This was confirmed through bacteriological identification and PCR as a field strain and smooth phenotypes. We conclude that seronegative cows vaccinated with RB51 which calved normally and shed B. abortus in the vaginal exudate after calving could be a serious problem because these cows are overlooked in routine diagnoses and are a source of Brucella infection.

  19. Comparative genomic analysis of Brucella abortus vaccine strain 104M reveals a set of candidate genes associated with its virulence attenuation.

    Science.gov (United States)

    Yu, Dong; Hui, Yiming; Zai, Xiaodong; Xu, Junjie; Liang, Long; Wang, Bingxiang; Yue, Junjie; Li, Shanhu

    2015-01-01

    The Brucella abortus strain 104M, a spontaneously attenuated strain, has been used as a vaccine strain in humans against brucellosis for 6 decades in China. Despite many studies, the molecular mechanisms that cause the attenuation are still unclear. Here, we determined the whole-genome sequence of 104M and conducted a comprehensive comparative analysis against the whole genome sequences of the virulent strain, A13334, and other reference strains. This analysis revealed a highly similar genome structure between 104M and A13334. The further comparative genomic analysis between 104M and A13334 revealed a set of genes missing in 104M. Some of these genes were identified to be directly or indirectly associated with virulence. Similarly, a set of mutations in the virulence-related genes was also identified, which may be related to virulence alteration. This study provides a set of candidate genes associated with virulence attenuation in B.abortus vaccine strain 104M.

  20. Immunogenicity of a Multi-Epitope DNA Vaccine Encoding Epitopes from Cu–Zn Superoxide Dismutase and Open Reading Frames of Brucella abortus in Mice

    Science.gov (United States)

    Escalona, Emilia; Sáez, Darwin; Oñate, Angel

    2017-01-01

    Brucellosis is a bacterial zoonotic disease affecting several mammalian species that is transmitted to humans by direct or indirect contact with infected animals or their products. In cattle, brucellosis is almost invariably caused by Brucella abortus. Live, attenuated Brucella vaccines are commonly used to prevent illness in cattle, but can cause abortions in pregnant animals. It is, therefore, desirable to design an effective and safer vaccine against Brucella. We have used specific Brucella antigens that induce immunity and protection against B. abortus. A novel recombinant multi-epitope DNA vaccine specific for brucellosis was developed. To design the vaccine construct, we employed bioinformatics tools to predict epitopes present in Cu–Zn superoxide dismutase and in the open reading frames of the genomic island-3 (BAB1_0260, BAB1_0270, BAB1_0273, and BAB1_0278) of Brucella. We successfully designed a multi-epitope DNA plasmid vaccine chimera that encodes and expresses 21 epitopes. This DNA vaccine induced a specific humoral and cellular immune response in BALB/c mice. It induced a typical T-helper 1 response, eliciting production of immunoglobulin G2a and IFN-γ particularly associated with the Th1 cell subset of CD4+ T cells. The production of IL-4, an indicator of Th2 activation, was not detected in splenocytes. Therefore, it is reasonable to suggest that the vaccine induced a predominantly Th1 response. The vaccine induced a statistically significant level of protection in BALB/c mice when challenged with B. abortus 2308. This is the first use of an in silico strategy to a design a multi-epitope DNA vaccine against B. abortus. PMID:28232837

  1. Oral immunization of mice with gamma-irradiated Brucella neotomae induces protection against intraperitoneal and intranasal challenge with virulent B. abortus 2308.

    Science.gov (United States)

    Dabral, Neha; Martha-Moreno-Lafont; Sriranganathan, Nammalwar; Vemulapalli, Ramesh

    2014-01-01

    Brucella spp. are Gram-negative, facultative intracellular coccobacilli that cause one of the most frequently encountered zoonosis worldwide. Humans naturally acquire infection through consumption of contaminated dairy and meat products and through direct exposure to aborted animal tissues and fluids. No vaccine against brucellosis is available for use in humans. In this study, we tested the ability of orally inoculated gamma-irradiated B. neotomae and B. abortus RB51 in a prime-boost immunization approach to induce antigen-specific humoral and cell mediated immunity and protection against challenge with virulent B. abortus 2308. Heterologous prime-boost vaccination with B. abortus RB51 and B. neotomae and homologous prime-boost vaccination of mice with B. neotomae led to the production of serum and mucosal antibodies specific to the smooth LPS. The elicited serum antibodies included the isotypes of IgM, IgG1, IgG2a, IgG2b and IgG3. All oral vaccination regimens induced antigen-specific CD4(+) and CD8(+) T cells capable of secreting IFN-γ and TNF-α. Upon intra-peritoneal challenge, mice vaccinated with B. neotomae showed the highest level of resistance against virulent B. abortus 2308 colonization in spleen and liver. Experiments with different doses of B. neotomae showed that all tested doses of 10(9), 10(10) and 10(11) CFU-equivalent conferred significant protection against the intra-peritoneal challenge. However, a dose of 10(11) CFU-equivalent of B. neotomae was required for affording protection against intranasal challenge as shown by the reduced bacterial colonization in spleens and lungs. Taken together, these results demonstrate the feasibility of using gamma-irradiated B. neotomae as an effective and safe oral vaccine to induce protection against respiratory and systemic infections with virulent Brucella.

  2. Seroprevalence for Coxiella burnetii, Francisella tularensis, Brucella abortus and Brucella melitensis in Austrian adults: a cross-sectional survey among military personnel and civilians.

    Science.gov (United States)

    Tobudic, Selma; Nedomansky, Klara; Poeppl, Wolfgang; Müller, Maria; Faas, Angelus; Mooseder, Gerhard; Allerberger, Franz; Stanek, Gerold; Burgmann, Heinz

    2014-04-01

    The prevalence of Coxiella burnetii, Francisella tularensis, Brucella abortus, and Brucella melitensis infections in Austria and the exposure risk of military personnel were assessed in an exploratory nationwide cross-sectional seroprevalence survey in 526 healthy adult individuals, 222 of which were soldiers and 304 were civilians. Screening for IgA/IgG antibodies to C. burnetii (Phase I) and IgG/IgM antibodies to C. burnetii (Phase II), and to F. tularensis was done with commercial enzyme-linked immunosorbent assays. To detect antibodies against B. abortus and B. melitensis, an in-house complement fixation test was used. Overall, 11 individuals (2.0%) showed antibodies to C. burnetii, 3 individuals (0.5%) were seropositive for F. tularensis, and one (0.3%) individual was borderline positive. All individuals positive or borderline for F. tularensis tested negative for antibodies against C. burnetii. All individuals tested negative for antibodies against B. melitensis/B. abortus. There were no significant differences between the seroprevalence of C. burnetii and F. tularensis among military personnel and civilians. Our data demonstrate serological evidence of a low rate of exposure to C. burnetii and F. tularensis among the Austrian adult population and military personnel.

  3. MLVA genotyping of Brucella melitensis and Brucella abortus isolates from different animal species and humans and identification of Brucella suis vaccine strain S2 from cattle in China.

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    Hai Jiang

    Full Text Available In China, brucellosis is an endemic disease and the main sources of brucellosis in animals and humans are infected sheep, cattle and swine. Brucella melitensis (biovars 1 and 3 is the predominant species, associated with sporadic cases and outbreak in humans. Isolates of B. abortus, primarily biovars 1 and 3, and B. suis biovars 1 and 3 are also associated with sporadic human brucellosis. In this study, the genetic profiles of B. melitensis and B. abortus isolates from humans and animals were analyzed and compared by multi-locus variable-number tandem-repeat analysis (MLVA. Among the B. melitensis isolates, the majority (74/82 belonged to MLVA8 genotype 42, clustering in the 'East Mediterranean' group. Two B. melitensis biovar 1 genotype 47 isolates, belonging to the 'Americas' group, were recovered; both were from the Himalayan blue sheep (Pseudois nayaur, a wild animal. The majority of B. abortus isolates (51/70 were biovar 3, genotype 36. Ten B. suis biovar 1 field isolates, including seven outbreak isolates recovered from a cattle farm in Inner Mongolia, were genetically indistinguishable from the vaccine strain S2, based on MLVA cluster analysis. MLVA analysis provided important information for epidemiological trace-back. To the best of our knowledge, this is the first report to associate Brucella cross-infection with the vaccine strain S2 based on molecular comparison of recovered isolates to the vaccine strain. MLVA typing could be an essential assay to improve brucellosis surveillance and control programs.

  4. Booster vaccination with safe, modified, live-attenuated mutants of Brucella abortus strain RB51 vaccine confers protective immunity against virulent strains of B. abortus and Brucella canis in BALB/c mice.

    Science.gov (United States)

    Truong, Quang Lam; Cho, Youngjae; Kim, Kiju; Park, Bo-Kyoung; Hahn, Tae-Wook

    2015-11-01

    Brucella abortus attenuated strain RB51 vaccine (RB51) is widely used in prevention of bovine brucellosis. Although vaccination with this strain has been shown to be effective in conferring protection against bovine brucellosis, RB51 has several drawbacks, including residual virulence for animals and humans. Therefore, a safe and efficacious vaccine is needed to overcome these disadvantages. In this study, we constructed several gene deletion mutants (ΔcydC, ΔcydD and ΔpurD single mutants, and ΔcydCΔcydD and ΔcydCΔpurD double mutants) of RB51 with the aim of increasing the safety of the possible use of these mutants as vaccine candidates. The RB51ΔcydC, RB51ΔcydD, RB51ΔpurD, RB51ΔcydCΔcydD and RB51ΔcydCΔpurD mutants exhibited significant attenuation of virulence when assayed in murine macrophages in vitro or in BALB/c mice. A single intraperitoneal immunization with RB51ΔcydC, RB51ΔcydD, RB51ΔcydCΔcydD or RB51ΔcydCΔpurD mutants was rapidly cleared from mice within 3 weeks, whereas the RB51ΔpurD mutant and RB51 were detectable in spleens until 4 and 7 weeks, respectively. Vaccination with a single dose of RB51 mutants induced lower protective immunity in mice than did parental RB51. However, a booster dose of these mutants provided significant levels of protection in mice against challenge with either the virulent homologous B. abortus strain 2308 or the heterologous Brucella canis strain 26. In addition, these mutants were found to induce a mixed but T-helper-1-biased humoral and cellular immune response in immunized mice. These data suggest that immunization with a booster dose of attenuated RB51 mutants provides an attractive strategy to protect against either bovine or canine brucellosis.

  5. Immune Response of Calves Vaccinated with Brucella abortus S19 or RB51 and Revaccinated with RB51

    Science.gov (United States)

    Dorneles, Elaine M. S.; Lima, Graciela K.; Teixeira-Carvalho, Andréa; Araújo, Márcio S. S.; Martins-Filho, Olindo A.; Sriranganathan, Nammalwar; Al Qublan, Hamzeh; Heinemann, Marcos B.; Lage, Andrey P.

    2015-01-01

    Brucella abortus S19 and RB51 strains have been successfully used to control bovine brucellosis worldwide; however, currently, most of our understanding of the protective immune response induced by vaccination comes from studies in mice. The aim of this study was to characterize and compare the immune responses induced in cattle prime-immunized with B. abortus S19 or RB51 and revaccinated with RB51. Female calves, aged 4 to 8 months, were vaccinated with either vaccine S19 (0.6–1.2 x 1011 CFU) or RB51 (1.3 x 1010 CFU) on day 0, and revaccinated with RB51 (1.3 x 1010 CFU) on day 365 of the experiment. Characterization of the immune response was performed using serum and peripheral blood mononuclear cells. Blood samples were collected on days 0, 28, 210, 365, 393 and 575 post-immunization. Results showed that S19 and RB51 vaccination induced an immune response characterized by proliferation of CD4+ and CD8+ T-cells; IFN-ɣ and IL-17A production by CD4+ T-cells; cytotoxic CD8+ T-cells; IL-6 secretion; CD4+ and CD8+ memory cells; antibodies of IgG1 class; and expression of the phenotypes of activation in T-cells. However, the immune response stimulated by S19 compared to RB51 showed higher persistency of IFN-ɣ and CD4+ memory cells, induction of CD21+ memory cells and higher secretion of IL-6. After RB51 revaccination, the immune response was chiefly characterized by increase in IFN-ɣ expression, proliferation of antigen-specific CD4+ and CD8+ T-cells, cytotoxic CD8+ T-cells and decrease of IL-6 production in both groups. Nevertheless, a different polarization of the immune response, CD4+- or CD8+-dominant, was observed after the booster with RB51 for S19 and RB51 prime-vaccinated animals, respectively. Our results indicate that after prime vaccination both vaccine strains induce a strong and complex Th1 immune response, although after RB51 revaccination the differences between immune profiles induced by prime-vaccination become accentuated. PMID:26352261

  6. Caspase-2-dependent dendritic cell death, maturation, and priming of T cells in response to Brucella abortus infection.

    Directory of Open Access Journals (Sweden)

    Xinna Li

    Full Text Available Smooth virulent Brucella abortus strain 2308 (S2308 causes zoonotic brucellosis in cattle and humans. Rough B. abortus strain RB51, derived from S2308, is a live attenuated cattle vaccine strain licensed in the USA and many other countries. Our previous report indicated that RB51, but not S2308, induces a caspase-2-dependent apoptotic and necrotic macrophage cell death. Dendritic cells (DCs are professional antigen presenting cells critical for bridging innate and adaptive immune responses. In contrast to Brucella-infected macrophages, here we report that S2308 induced higher levels of apoptotic and necrotic cell death in wild type bone marrow-derived DCs (WT BMDCs than RB51. The RB51 and S2308-induced BMDC cell death was regulated by caspase-2, indicated by the minimal cell death in RB51 and S2308-infected BMDCs isolated from caspase-2 knockout mice (Casp2KO BMDCs. More S2308 bacteria were taken up by Casp2KO BMDCs than wild type BMDCs. Higher levels of S2308 and RB51 cells were found in infected Casp2KO BMDCs compared to infected WT BMDCs at different time points. RB51-infected wild type BMDCs were mature and activated as shown by significantly up-regulated expression of CD40, CD80, CD86, MHC-I, and MHC-II. RB51 induced the production of cytokines TNF-α, IL-6, IFN-γ and IL12/IL23p40 in infected BMDCs. RB51-infected WT BMDCs also stimulated the proliferation of CD4(+ and CD8(+ T cells compared to uninfected WT BMDCs. However, the maturation, activation, and cytokine secretion are significantly impaired in Casp2KO BMDCs infected with RB51 or Salmonella (control. S2308-infected WT and Casp2KO BMDCs were not activated and could not induce cytokine production. These results demonstrated that virulent smooth strain S2308 induced more apoptotic and necrotic dendritic cell death than live attenuated rough vaccine strain RB51; however, RB51, but not its parent strain S2308, induced caspase-2-mediated DC maturation, cytokine production, antigen

  7. Immune Response of Calves Vaccinated with Brucella abortus S19 or RB51 and Revaccinated with RB51.

    Science.gov (United States)

    Dorneles, Elaine M S; Lima, Graciela K; Teixeira-Carvalho, Andréa; Araújo, Márcio S S; Martins-Filho, Olindo A; Sriranganathan, Nammalwar; Al Qublan, Hamzeh; Heinemann, Marcos B; Lage, Andrey P

    2015-01-01

    Brucella abortus S19 and RB51 strains have been successfully used to control bovine brucellosis worldwide; however, currently, most of our understanding of the protective immune response induced by vaccination comes from studies in mice. The aim of this study was to characterize and compare the immune responses induced in cattle prime-immunized with B. abortus S19 or RB51 and revaccinated with RB51. Female calves, aged 4 to 8 months, were vaccinated with either vaccine S19 (0.6-1.2 x 1011 CFU) or RB51 (1.3 x 1010 CFU) on day 0, and revaccinated with RB51 (1.3 x 1010 CFU) on day 365 of the experiment. Characterization of the immune response was performed using serum and peripheral blood mononuclear cells. Blood samples were collected on days 0, 28, 210, 365, 393 and 575 post-immunization. Results showed that S19 and RB51 vaccination induced an immune response characterized by proliferation of CD4+ and CD8+ T-cells; IFN-ɣ and IL-17A production by CD4+ T-cells; cytotoxic CD8+ T-cells; IL-6 secretion; CD4+ and CD8+ memory cells; antibodies of IgG1 class; and expression of the phenotypes of activation in T-cells. However, the immune response stimulated by S19 compared to RB51 showed higher persistency of IFN-ɣ and CD4+ memory cells, induction of CD21+ memory cells and higher secretion of IL-6. After RB51 revaccination, the immune response was chiefly characterized by increase in IFN-ɣ expression, proliferation of antigen-specific CD4+ and CD8+ T-cells, cytotoxic CD8+ T-cells and decrease of IL-6 production in both groups. Nevertheless, a different polarization of the immune response, CD4+- or CD8+-dominant, was observed after the booster with RB51 for S19 and RB51 prime-vaccinated animals, respectively. Our results indicate that after prime vaccination both vaccine strains induce a strong and complex Th1 immune response, although after RB51 revaccination the differences between immune profiles induced by prime-vaccination become accentuated.

  8. Seroprevalence survey of Chlamydophila abortus infection in breeding goats on commercial farms in the Otavi Veterinary District, northern Namibia

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    Alaster Samkange

    2010-08-01

    Full Text Available A total of 1 076 sera from breeding goats were randomly collected from 24 different farms and tested with CHEKIT®-ELISA (IDEXX Laboratories B.V., 1 119 NE Schiphol-Rijk, Nederland for antibodies against Chlamydophila abortus. The farms were divided into two categories of twelve farms each,based on their previous history of observed abortions over the previous 12 months: those with low (< 5% levels of abortion and those with high (≥ 5% levels of abortion. The farmers were also interviewed on their level of awareness about chlamydophilosis, its zoonotic importance and vaccination measures against the disease. The study detected overall seroprevalence levels of 25% for the farms and 8% for the individual animals (at 95% confidence. A total of six out of twentyfour farms (25% had at least one positive breeding animal. Only five out of the twenty-four (20.8%farmers interviewed were aware of chlamydophilosis and its zoonotic dangers. None of the 24 farmers interviewed practised any vaccination against chlamydophilosis. There was a significantly higher number of seropositive animals from farms with high levels of abortion, compared to those animals from farms with low levels of abortion (p = 0.0001. This study underscores the need for a higher level of farmer awareness and training on chlamydophilosis and its zoonotic dangers.

  9. Reaction of Native and Denatured Brucella abortus (S19 Proteins with Antibody Using Affinity Chromatography and Immunoblotting

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    R. Karimi

    2005-01-01

    Full Text Available Western blotting or immunoblotting commonly use for study of reaction between antigens and antibodies. Denaturation of many proteins in immunoblotting can affect greatly the reactivity of antibodies and outcome of the procedure.In this study proteins of Brucella abortus (S19 was extracted by a mild method and reaction of the extracted proteins with serum of infected human and goat and immunized rabbit compared by affinity chromatography and immunoblotting. Gamma globulin (mostly IgG fraction of the sera was precipitated by half saturation of ammonium sulfate and linked to activated sepharose 4B. The extracted proteins were loaded on the affinity column. Attached proteins was eluted by low pH and analyzed by SDS-PAGE. Reaction of the total extract and eluted fractions with IgG fraction of sera was evaluated by Western blotting.Upon the results of affinity chromatography and immunoblotting, Brucella proteins can be classified in four groups: 1- The proteins that adsorbed to the affinity column and react with IgG in westernblotting. 2- Proteins that react with IgG in native state but no in denatured state. 3- Proteins that do not react with IgG in native state but react in denatured state. 4- Proteins that do not react with IgG in native and denatured state.

  10. The unexpected discovery of Brucella abortus Buck 19 vaccine in goats from Ecuador underlines the importance of biosecurity measures.

    Science.gov (United States)

    Ron-Román, Jorge; Berkvens, Dirk; Barzallo-Rivadeneira, Daniela; Angulo-Cruz, Alexandra; González-Andrade, Pablo; Minda-Aluisa, Elizabeth; Benítez-Ortíz, Washington; Brandt, Jef; Rodríguez-Hidalgo, Richar; Saegerman, Claude

    2017-03-01

    Very few, mostly old, and only preliminary serological studies of brucellosis in goats exist in Ecuador. In order to assess the current epidemiological situation, we performed a cross-sectional serological study in the goat populations of Carchi (n = 160 animals), Pichincha (n = 224 animals), and Loja provinces (n = 2024 animals). Only two positive serological results (RB negative and SAT-EDTA ≥400 IU/ml) were obtained in lactating goats from the same farm in Quito (Pichincha province). Additionally, milk was sampled from 220 animals in Pichincha province. The present study indicates a low apparent prevalence in Pichincha province and absence in Carchi and Loja provinces. A total of 25 positive milk ring tests (MRT) were obtained in Pichincha province yielding a prevalence of MRT of 11.16%. Subsequent culture was performed on the positive MRT samples. All results were negative, apart from a single sample, obtained from a serologically positive goat in Quito, that was positive for Brucella abortus strain 19 (B19). Several hypotheses are forwarded concerning this unexpected result. The most likely hypothesis is the possible accidental use of a needle, previously used for vaccination of cattle with the said vaccine, for the administration of drug treatment to the goat. This hypothesis underlines the necessity of biosecurity measures to prevent this type of accidents.

  11. Seroprevalence survey of Chlamydophila abortus infection in breeding goats on commercial farms in the Otavi Veterinary District, northern Namibia.

    Science.gov (United States)

    Samkange, Alaster; Katsande, Tendai C; Tjipura-Zaire, Georgina; Crafford, Jan E

    2010-08-20

    A total of 1 076 sera from breeding goats were randomly collected from 24 different farms and tested with CHEKIT®-ELISA (IDEXX Laboratories B.V., 1 119 NE Schiphol-Rijk, Nederland) for antibodies against Chlamydophila abortus. The farms were divided into two categories of twelve farms each,based on their previous history of observed abortions over the previous 12 months: those with low (< 5%) levels of abortion and those with high (≥ 5%) levels of abortion. The farmers were also interviewed on their level of awareness about chlamydophilosis, its zoonotic importance and vaccination measures against the disease. The study detected overall seroprevalence levels of 25% for the farms and 8% for the individual animals (at 95% confidence). A total of six out of twentyfour farms (25%) had at least one positive breeding animal. Only five out of the twenty-four (20.8%)farmers interviewed were aware of chlamydophilosis and its zoonotic dangers. None of the 24 farmers interviewed practised any vaccination against chlamydophilosis. There was a significantly higher number of seropositive animals from farms with high levels of abortion, compared to those animals from farms with low levels of abortion (p = 0.0001). This study underscores the need for a higher level of farmer awareness and training on chlamydophilosis and its zoonotic dangers.

  12. Subcutaneous immunization with a novel immunogenic candidate (urease) confers protection against Brucella abortus and Brucella melitensis infections.

    Science.gov (United States)

    Abkar, Morteza; Amani, Jafar; Sahebghadam Lotfi, Abbas; Nikbakht Brujeni, Gholamreza; Alamian, Saeed; Kamali, Mehdi

    2015-08-01

    Brucellosis is a world prevalent endemic illness that is transmitted from domestic animals to humans. Brucella spp. exploits urease for survival in the harsh conditions of stomach during the gastrointestinal infection. In this study, we examined the immune response and the protection elicited by using recombinant Brucella urease (rUrease) vaccination in BALB/c mice. The urease gene was cloned in pET28a and the resulting recombinant protein was employed as subunit vaccine. Recombinant protein was administered subcutaneously and intraperitoneally. Dosage reduction was observed with subcutaneous (SC) vaccination when compared with intraperitoneal (IP) vaccination. rUrease induced mixed Th1-Th2 immune responses with high titers of specific IgG1 and IgG2a. In lymphocyte proliferation assay, splenocytes from IP and SC-vaccinated mice displayed a strong recall proliferative response with high amounts of IL-4, IL-12 and IFN-γ production. Vaccinated mice were challenged with virulent Brucella melitensis, B. abortus and B. suis. The SC vaccination route exhibited a higher degree of protection than IP vaccination (p value ≤ 0.05). Altogether, our results indicated that rUrease could be a useful antigen candidate for the development of subunit vaccines against brucellosis.

  13. SodA is a major metabolic antioxidant in Brucella abortus 2308 that plays a significant, but limited, role in the virulence of this strain in the mouse model.

    Science.gov (United States)

    Martin, Daniel W; Baumgartner, John E; Gee, Jason M; Anderson, Eric S; Roop, R Martin

    2012-07-01

    The gene designated BAB1_0591 in the Brucella abortus 2308 genome sequence encodes the manganese-cofactored superoxide dismutase SodA. An isogenic sodA mutant derived from B. abortus 2308, designated JB12, displays a small colony phenotype, increased sensitivity in vitro to endogenous superoxide generators, hydrogen peroxide and exposure to acidic pH, and a lag in growth when cultured in rich and minimal media that can be rescued by the addition of all 20 amino acids to the growth medium. B. abortus JB12 exhibits significant attenuation in both cultured murine macrophages and experimentally infected mice, but this attenuation is limited to the early stages of infection. Addition of the NADPH oxidase inhibitor apocynin to infected macrophages does not alleviate the attenuation exhibited by JB12, suggesting that the basis for the attenuation of the B. abortus sodA mutant is not an increased sensitivity to exogenous superoxide generated through the oxidative burst of host phagocytes. It is possible, however, that the increased sensitivity of the B. abortus sodA mutant to acid makes it less resistant than the parental strain to killing by the low pH encountered during the early stages of the development of the brucella-containing vacuoles in macrophages. These experimental findings support the proposed role for SodA as a major cytoplasmic antioxidant in brucella. Although this enzyme provides a clear benefit to B. abortus 2308 during the early stages of infection in macrophages and mice, SodA appears to be dispensable once the brucellae have established an infection.

  14. The Attenuated Brucella abortus Strain 19 Invades, Persists in, and Activates Human Dendritic Cells, and Induces the Secretion of IL-12p70 but Not IL-23

    Science.gov (United States)

    Weinhold, Mario; Eisenblätter, Martin; Jasny, Edith; Fehlings, Michael; Finke, Antje; Gayum, Hermine; Rüschendorf, Ursula; Renner Viveros, Pablo; Moos, Verena; Allers, Kristina; Schneider, Thomas; Schaible, Ulrich E.; Schumann, Ralf R.; Mielke, Martin E.; Ignatius, Ralf

    2013-01-01

    Background Bacterial vectors have been proposed as novel vaccine strategies to induce strong cellular immunity. Attenuated strains of Brucella abortus comprise promising vector candidates since they have the potential to induce strong CD4+ and CD8+ T-cell mediated immune responses in the absence of excessive inflammation as observed with other Gram-negative bacteria. However, some Brucella strains interfere with the maturation of dendritic cells (DCs), which is essential for antigen-specific T-cell priming. In the present study, we investigated the interaction of human monocyte-derived DCs with the smooth attenuated B. abortus strain (S) 19, which has previously been employed successfully to vaccinate cattle. Methodology/Principal findings We first looked into the potential of S19 to hamper the cytokine-induced maturation of DCs; however, infected cells expressed CD25, CD40, CD80, and CD86 to a comparable extent as uninfected, cytokine-matured DCs. Furthermore, S19 activated DCs in the absence of exogeneous stimuli, enhanced the expression of HLA-ABC and HLA-DR, and was able to persist intracellularly without causing cytotoxicity. Thus, DCs provide a cellular niche for persisting brucellae in vivo as a permanent source of antigen. S19-infected DCs produced IL-12/23p40, IL-12p70, and IL-10, but not IL-23. While heat-killed bacteria also activated DCs, soluble mediators were not involved in S19-induced activation of human DCs. HEK 293 transfectants revealed cellular activation by S19 primarily through engagement of Toll-like receptor (TLR)2. Conclusions/Significance Thus, as an immunological prerequisite for vaccine efficacy, B. abortus S19 potently infects and potently activates (most likely via TLR2) human DCs to produce Th1-promoting cytokines. PMID:23805193

  15. Investigations concerning the prevalence of Coxiella burnetii and Chlamydia abortus in sheep in correlation with management systems and abortion rate in Lower Saxony in 2004.

    Science.gov (United States)

    Runge, Martin; Binder, Alfred; Schotte, Ulrich; Ganter, Martin

    2012-01-01

    The intracellular bacteria Coxiella (C) burnetii and Chlamydia (Chl) abortus induce abortion in sheep and also affect humans. While Chl. abortus only infrequently infects humans, C burnetii is the aetiological agent of numerous Q fever outbreaks during the last decades. There is only limited knowledge about the prevalence of both pathogens in sheep, although sheep are involved in almost all Q fever outbreaks in Germany. The aim of our study was to investigate the prevalence of both pathogens in flocks located in Lower Saxony, Germany, in correlation to the management form and abortion rate. Serum samples of 1714 sheep from 95 flocks located in Lower Saxony were investigated by ELISA. 2.7% of these samples were positive, 1.3% showed inconclusive results in the C. burnetii-ELISA. Elevated intra-flock seroprevalences were only detected in three migrating flocks. Chlamydia-specific antibodies could be detected in 15.1% serum samples of mainly shepherded and migrating flocks. In one of these flocks with a high intra-flock seroprevalence for C burnetii (27%) and Chlamydia (44.9%), C burnetii was detected in 21.6% of the placenta samples of normal births and in 12.5% of the colostrum samples by PCR. Aborted fetuses and the corresponding placentas were negative in C burnetii-PCR, but in most of them and also in many other placenta samples Chl. abortus could be detected by PCR and DNA microarray. This survey shows a low overall prevalence of C. burnetii in sheep in Lower Saxony in the year 2004. However, three migrating flocks with a high intra-flock prevalence are localized in the southern parts of Lower Saxony. Spreading of C burnetii could occur, because of the large radius of grazing of all three flocks.

  16. Critical role of ASC inflammasomes and bacterial type IV secretion system in caspase-1 activation and host innate resistance to Brucella abortus infection.

    Science.gov (United States)

    Gomes, Marco Tulio R; Campos, Priscila C; Oliveira, Fernanda S; Corsetti, Patricia P; Bortoluci, Karina R; Cunha, Larissa D; Zamboni, Dario S; Oliveira, Sergio C

    2013-04-01

    Pathogens are detected by innate immune receptors that, upon activation, orchestrate an appropriate immune response. Recent studies revealed the intracellular signaling cascades involved in the TLR-initiated immune response to Brucella abortus infection. However, no report has elucidated the role of inflammasome receptors in Brucella recognition. Therefore, we decided to investigate the function of NLRC4, NLRP3, and AIM2 in sensing Brucella. In this study, we showed that NLRC4 is not required to induce caspase-1 activation and further secretion of IL-1β by B. abortus in macrophages. In contrast, we determined that AIM2, which senses Brucella DNA, and NLRP3 are partially required for caspase-1 activation and IL-1β secretion. Additionally, mitochondrial reactive oxygen species induced by Brucella were implicated in IL-1β production. Furthermore, AIM2, NLRP3, ASC, and caspase-1 knockout mice were more susceptible to B. abortus infection than were wild-type animals, suggesting that multiple ASC-dependent inflammasomes contribute to host protection against infection. This protective effect is due to the inflammatory response caused by IL-1β and IL-18 rather than pyroptosis, because we observed augmented bacterial burden in IL-1R and IL-18 knockout mice. Finally, we determined that bacterial type IV secretion system VirB and live, but not heat-killed, Brucella are required for full inflammasome activation in macrophages during infection. Taken together, our results indicate that Brucella is sensed by ASC inflammasomes that collectively orchestrate a robust caspase-1 activation and proinflammatory response.

  17. The Attenuated Brucella abortus Strain 19 Invades, Persists in, and Activates Human Dendritic Cells, and Induces the Secretion of IL-12p70 but Not IL-23.

    Directory of Open Access Journals (Sweden)

    Mario Weinhold

    Full Text Available Bacterial vectors have been proposed as novel vaccine strategies to induce strong cellular immunity. Attenuated strains of Brucella abortus comprise promising vector candidates since they have the potential to induce strong CD4(+ and CD8(+ T-cell mediated immune responses in the absence of excessive inflammation as observed with other Gram-negative bacteria. However, some Brucella strains interfere with the maturation of dendritic cells (DCs, which is essential for antigen-specific T-cell priming. In the present study, we investigated the interaction of human monocyte-derived DCs with the smooth attenuated B. abortus strain (S 19, which has previously been employed successfully to vaccinate cattle.We first looked into the potential of S19 to hamper the cytokine-induced maturation of DCs; however, infected cells expressed CD25, CD40, CD80, and CD86 to a comparable extent as uninfected, cytokine-matured DCs. Furthermore, S19 activated DCs in the absence of exogeneous stimuli, enhanced the expression of HLA-ABC and HLA-DR, and was able to persist intracellularly without causing cytotoxicity. Thus, DCs provide a cellular niche for persisting brucellae in vivo as a permanent source of antigen. S19-infected DCs produced IL-12/23p40, IL-12p70, and IL-10, but not IL-23. While heat-killed bacteria also activated DCs, soluble mediators were not involved in S19-induced activation of human DCs. HEK 293 transfectants revealed cellular activation by S19 primarily through engagement of Toll-like receptor (TLR2.Thus, as an immunological prerequisite for vaccine efficacy, B. abortus S19 potently infects and potently activates (most likely via TLR2 human DCs to produce Th1-promoting cytokines.

  18. Prueba de inmunodifusión radial con hapteno nativo para diferenciar bovinos con revacunaciones repetidas con la cepa S19 de Brucella abortus

    OpenAIRE

    Esperanza González Miranda; Laura Hernández Andrade; Efrén Díaz Aparicio

    2006-01-01

    El objetivo del estudio fue establecer la capacidad de la inmunodifusión radial (IDR) con hapteno nativo, para diferenciar anticuerpos post-vacunales de anticuerpos por infección, en bovinos con revacunaciones repetidas de cepa S19. Se trabajó en un establo del estado de México, con presencia de abortos, retención de placenta, expulsiones y fetos momificados. Las becerras se vacunaban con dosis clásica de cepa S19 de B. abortus, y ya adultas revacunaciones anuales repetidas con dosis reducida...

  19. Aglutininas anti-Brucella abortus no soro e em secreção de bursite cervical em eqüinos

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    Ribeiro M.G.

    2003-01-01

    Full Text Available Fistulous wither secretions from three horses were tested by the plate agglutination (PAT, tube agglutination (SAT, buffered plate-Rose Bengal (RBPT and 2-mercaptoethanol (2-ME tests, comparatively with standard agglutination tests. In the modified tests, titers were increased in the PAT, SAT and 2-ME tests and positive reaction was observed in RBPT. Brucella abortus was isolated from the secretion of fistulous withers collected from one animal. These results suggest that the modified tests may be used as alternative tests to diagnose brucellosis in horses with fistulous withers.

  20. Express immunochromatographic detection of antibodies against Brucella abortus in cattle sera based on quantitative photometric registration and modulated cut-off level.

    Science.gov (United States)

    Sotnikov, Dmitriy V; Byzova, Nadezhda A; Zherdev, Anatoly V; Eskendirova, Saule Z; Baltin, Kairat K; Mukanov, Kasim K; Ramankulov, Erlan M; Sadykhov, Elchin G; Dzantiev, Boris B

    2015-01-01

    An immunochromatographic test system was developed for rapid detection of the levels of specific IgG antibodies to Brucella abortus lipopolysaccharide, as a tool for diagnosis of brucellosis in cattle. The pilot test strips were examined using blood sera from sick (78 samples) and healthy (35 samples) cows. The results obtained by immunochromatographic assay, using a portable optical densitometer for digital video detection, correlate well with the results obtained by immunoenzyme assay and are in agreement with the results of the disease diagnosis. The new test system allows detection of antibodies within 10 min and can be proposed as an alternative to the methods available for serodiagnosis of brucellosis.

  1. Efecto de una dieta con bajo aporte de selenio sobre la respuesta inmune a la vacuna Brucella abortus Cepa RB51 en vacas lecheras Effect of a low selenium diet on the immune response to Brucella abortus strain RB51 vaccine in dairy cows

    Directory of Open Access Journals (Sweden)

    V Leyán

    2006-01-01

    Full Text Available Se estudió el efecto de una dieta con bajo aporte de selenio (Se sobre la respuesta inmune a la vacuna Brucella abortus Cepa RB51 y la concentración de inmunoglobulinas séricas en vacas. Se utilizaron 12 vacas Friesian, estabuladas desde aproximadamente dos meses preparto y hasta el cuarto mes de lactancia mantenidas con una dieta basada en heno de pradera con bajo contenido de Se (0,02 ppm de MS y balanceada según requerimientos para el resto de nutrientes. Seis vacas conformaron el grupo de animales con bajo aporte de Se (Se-D y otras seis el grupo de animales suplementados (Se-S con selenato de bario (1 mg de Se/kg , 45 días previos al parto. Los animales fueron inmunizados con la vacuna RB51 al cuarto mes del experimento. Muestras de sangre fueron obtenidas previo a la suplementación con Se y cada 15 días hasta el término del experimento. El balance de Se fue medido mediante la actividad sanguínea de GSH-Px. Las concentraciones séricas de IgG, IgM e IgA se determinaron por inmunodifusión y los anticuerpos específicos contra Brucella abortus mediante ELISA y la respuesta inmune celular mediante pruebas de intradermorreacción a antígenos de Brucella abortus y estudio histológico de la reacción. La dieta con bajo contenido de Se provocó una disminución lenta y progresiva de la actividad de GSH-Px (The effect of a diet with a low selenium (Se content on the immune response to Brucella abortus Strain RB51 vaccine in dairy cows and in their serum inmunoglobulin concentrations was studied. Twelve pregnant Friesian cows (7 to 8 months were randomly allocated into two homogeneous groups of six animals each. Animals were maintained during 6 months in individual cubicles with water ad libitum and a diet based on grass hay with a low Se content (0.02 ppm base on dry matter and nutritionally balanced for other nutrients. One group was maintained only with the low Se diet (Se-D and the other group (Se-S was treated with barium selenate

  2. Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 Avaliação de diferentes protocolos de extração de DNA para detecção de Brucella abortus a partir de abortos ou de bezerros nascidos de vacas experimentalmente infectadas com estirpe 2308

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    M. Matrone

    2009-09-01

    Full Text Available The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives, 26 spleens (11 positives, 23 livers (8 positives and 22 bronchial lymph nodes (7 positives. All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04 or bronchial lymph nodes (p=0.004 and equal to the spleens (p=0.18. From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (pO objetivo do presente estudo foi aperfeiçoar a detecção de Brucella abortus pela PCR em homogeneizados de órgãos de fetos abortados por vacas infectadas, importante mecanismo para descobrir focos da doença na fase de erradicação. Assim, foram comparados diferentes protocolos de extração de DNA, visando à detecção de B. abortus pela PCR em amostras clínicas colhidas de abortos ou de bezerros oriundos de vacas desafiadas com a estirpe 2308 de B. abortus. Para tanto, foram construídos dois grupos padrão ouro com base na bacteriologia clássica, constituídos por: 32 pulmões (17 positivos, 26 baços (11 positivos, 23 fígados (8 positivos e 22 linfonodos bronquiais (7 positivos. Todas essas amostras foram submetidas a três protocolos de extração de DNA, seguidos do mesmo processo de amplificação com os primers B4 e B5. Nos resultados acumulados por

  3. Evaluation of four DNA extraction protocols for Brucella abortus detection by PCR in tissues from experimentally infected cows with the 2308 strain.

    Science.gov (United States)

    Vejarano, M P; Matrone, M; Keid, L B; Rocha, V C M; Ikuta, C Y; Rodriguez, C A R; Salgado, V R; Ferreira, F; Dias, R A; Telles, E O; Ferreira Neto, J S

    2013-04-01

    This study compared 4 protocols for DNA extraction from homogenates of 6 different organs of cows infected with the Brucella abortus 2308 strain. The extraction protocols compared were as follows: GT (guanidine isothiocyanate lysis), Boom (GT lysis with the carrying suspension diatomaceous earth), PK (proteinase K lysis), and Santos (lysis by boiling and freezing with liquid nitrogen). Positive and negative gold standard reference groups were generated by classical bacteriological methods. All samples were processed with the 4 DNA extraction protocols and amplified with the B4 and B5 primers. The number of positive samples in the placental cotyledons was higher than that in the other organs. The cumulated results showed that the Santos protocol was more sensitive than the Boom (p=0.003) and GT (p=0.0506) methods and was similar to the PK method (p=0.2969). All of the DNA extraction protocols resulted in false-negative results for PCR. In conclusion, despite the disadvantages of classical bacteriological methods, the best approach for direct diagnosis of B. abortus in organs of infected cows includes the isolation associated with PCR of DNA extracted from the cotyledon by the Santos or PK methods.

  4. Epidemiological aspects of an infection by Brucella abortus in risk occupational groups in the microregion of Araguaína, Tocantins

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    Taciana Rabelo Ramalho Ramos

    2008-04-01

    Full Text Available The aim of this paper was to study some epidemiological aspects of the infection by Brucella abortus in risk occupational groups in the microregion of Araguaína, Tocantins. For antibody research, 645 serum samples were analyzed by the complement fixation test (CF. A 4.0% frequency was found (26/645 in patients' serum and among those 4.1% (23/551 were slaughterhouses employees and 8.1% (3/37 rural workers. Of the total positive samples, three (2.0% were women and 23 (4.7% men; ten (2.9% were between the ages of 18 and 30, six (3.4% between 31 and 40, and nine (8.0% were above 41 years of age. Risk factors for brucellosis in the study groups were age, background (OR = 2.45; CI 95% = 0.98 to 6.10 and previous work conducted with production animals (OR 2.36; CI 95% = 0.95 to 6.02. It was concluded that the infection by Brucella abortus is found in some risk occupational groups in the microregion of Araguaína, Tocantins, and control and prophylactic measures must be implemented emphasizing risk factors identified in the study.

  5. Vaccination of adult animals with a reduced dose of Brucella abortus S19 vaccine to control brucellosis on dairy farms in endemic areas of India.

    Science.gov (United States)

    Chand, Puran; Chhabra, Rajesh; Nagra, Juhi

    2015-01-01

    Bovine brucellosis is an economically important disease which seriously affects dairy farming by causing colossal losses. It can be controlled by practicing vaccination of animals with Brucella abortus S19 vaccine (S19 vaccine). In the present study, adult bovines were vaccinated on seven dairy farms with a reduced dose of S19 vaccine to control brucellosis. Serological screening of adult animals (N = 1,082) by Rose Bengal test (RBT) and ELISA prior to vaccination revealed the presence and absence of brucellosis on five and two farms, respectively. The positive animals (N = 171) were segregated and those which tested negative (N = 911) were vaccinated by conjunctival route with a booster after 4 months. The conjunctival vaccination induced weak antibody response in animals, which vanished within a period of 9 to 12 weeks. Abortion in 12 animals at various stages of pregnancy and post-vaccination was recorded, but none was attributed to S19 vaccine. However, virulent B. abortus was incriminated in six heifers, and the cause of abortion could not be established in six animals. The six aborted heifers perhaps acquired infection through in utero transmission or from the environment which remained undetected until abortion. These findings suggested that vaccination of adult animals with a reduced dose of S19 vaccine by conjunctival route did not produce adverse effects like abortion in pregnant animals and persistent vaccinal antibody titers, which are the major disadvantages of subcutaneous vaccination of adult animals.

  6. Intermediate rough Brucella abortus S19Δper mutant is DIVA enable, safe to pregnant guinea pigs and confers protection to mice.

    Science.gov (United States)

    Lalsiamthara, Jonathan; Gogia, Neha; Goswami, Tapas K; Singh, R K; Chaudhuri, Pallab

    2015-05-21

    Brucella abortus S19 is a smooth strain used as live vaccine against bovine brucellosis. Smooth lipopolysaccharide (LPS) is responsible for its residual virulence and serological interference. Rough mutants defective of LPS are more attenuated but confers lower level of protection. We describe a modified B. abortus S19 strain, named as S19Δper, which exhibits intermediate rough phenotype with residual O-polysaccharide (OPS). Deletion of perosamine synthetase gene resulted in substantial attenuation of S19Δper mutant without affecting immunogenic properties. It mounted strong immune response in Swiss albino mice and conferred protection similar to S19 vaccine. Immunized mice produced higher levels of IFN-γ, IgG2a and thus has immune response inclined towards Th1 cell mediated immunity. Sera from immunized animals did not show agglutination reaction with RBPT antigen and thus could serve as DIVA (Differentiating Infected from Vaccinated Animals) vaccine. S19Δper mutant displayed more susceptibility to serum complement mediated killing and sensitivity to polymyxin B. Pregnant guinea pigs injected with S19Δper mutant completed full term of pregnancy and did not cause abortion, still birth or birth of weak offspring. S19Δper mutant with intermediate rough phenotype displayed remarkable resemblance to S19 vaccine strain with improved properties of safety, immunogenicity and DIVA capability for control of bovine brucellosis.

  7. MyD88 and STING signaling pathways are required for IRF3-mediated IFN-β induction in response to Brucella abortus infection.

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    Leonardo A de Almeida

    Full Text Available Type I interferons (IFNs are cytokines that orchestrate diverse immune responses to viral and bacterial infections. Although typically considered to be most important molecules in response to viruses, type I IFNs are also induced by most, if not all, bacterial pathogens. In this study, we addressed the role of type I IFN signaling during Brucella abortus infection, a facultative intracellular bacterial pathogen that causes abortion in domestic animals and undulant fever in humans. Herein, we have shown that B. abortus induced IFN-β in macrophages and splenocytes. Further, IFN-β induction by Brucella was mediated by IRF3 signaling pathway and activates IFN-stimulated genes via STAT1 phosphorylation. In addition, IFN-β expression induced by Brucella is independent of TLRs and TRIF signaling but MyD88-dependent, a pathway not yet described for Gram-negative bacteria. Furthermore, we have identified Brucella DNA as the major bacterial component to induce IFN-β and our study revealed that this molecule operates through a mechanism dependent on RNA polymerase III to be sensed probably by an unknown receptor via the adaptor molecule STING. Finally, we have demonstrated that IFN-αβR KO mice are more resistant to infection suggesting that type I IFN signaling is detrimental to host control of Brucella. This resistance phenotype is accompanied by increased IFN-γ and NO production by IFN-αβR KO spleen cells and reduced apoptosis.

  8. The protoxin Cry1Ac of Bacillus thuringiensis improves the protection conferred by intranasal immunization with Brucella abortus RB51 in a mouse model.

    Science.gov (United States)

    González-González, Edith; García-Hernández, Ana Lilia; Flores-Mejía, Raúl; López-Santiago, Rubén; Moreno-Fierros, Leticia

    2015-02-25

    Brucellosis is a zoonotic disease affecting many people and animals worldwide. Preventing this infection requires improving vaccination strategies. The protoxin Cry1Ac of Bacillus thuringiensis is an adjuvant that, in addition to increasing the immunogenicity of different antigens, has shown to be protective in different models of parasitic infections. The objective of the present study was to test whether the intranasal co-administration of pCry1Ac with the RB51 vaccine strain of Brucella abortus confers protection against an intranasal challenge with the virulent strain B. abortus 2308 in BALB/c mice. The results showed that co-administration of pCry1Ac and RB51, increased the immunoprotection conferred by the vaccine as evidenced by the following: (1) decrease of the splenic bacterial load when challenged intranasally with the virulent strain; (2) greater in vivo cytotoxic activity in response to the transference of previously infected cells; (3) further proliferation of cytotoxic TCD8+ cells in response to stimulation with heat-inactivated bacteria; (4) increased production of TNF-α and IFN-γ; and (5) significant IgG2a response. These results indicate that the use of the Cry1Ac protein as a mucosal adjuvant via the intranasal route can be a promising alternative for improving current RB51 vaccine against brucellosis.

  9. A diagnostic protocol to identify water buffaloes (Bubalus bubalis) vaccinated with Brucella abortus strain RB51 vaccine.

    Science.gov (United States)

    Tittarelli, Manuela; Atzeni, Marcello; Calistri, Paolo; Di Giannatale, Elisabetta; Ferri, Nicola; Marchi, Enrico; Martucciello, Alessandra; De Massis, Fabrizio

    2015-01-01

    The use of live vaccine strain RB51 for vaccination of domestic water buffaloes (Bubalus bubalis) at risk of infection with Brucella abortus is permitted notwithstanding the plans for the eradication and only under strict veterinary control. The antibodies induced by RB51 vaccination are not detectable using conventional diagnostic techniques; therefore, it is necessary to have a specific diagnostic tool able to discriminate vaccinated from unvaccinated animals. The combination of a complement fixation test (CFT) with specific RB51 antigen (RB51-CFT) and a brucellin skin test has been demonstrated to be a reliable diagnostic system to identify single cattle (Bos taurus) vaccinated with RB51. So far, no data are available in the international scientific literature regarding the use of this test association in water buffalo. For this reason the suitability of this test combination has been evaluated in a water buffalo herd. One hundred twenty-seven animals farmed in a herd of Salerno province (Campania, Southern Italy), in the context of a presumptive unauthorized use of RB51 vaccine were chosen for this study. All tested animals resulted negative to Rose Bengal test (RBT) and complement fixation test (CFT) used for the detection of specific antibodies against Brucella field strains. Seventy-one animals (56%) developed RB51 antigen-specific CFT (RB51-CFT) antibodies against RB51 vaccine in a first sampling, while 104 animals (82%) gave positive result to a second serum sampling conducted 11 days after the intradermal inoculation of the RB51 brucellin. One hundred and seven animals (84%) showed a positive reaction to the RB51-CFT in at least 1 sampling, while 111 animals (87%) resulted positive to the RB51 brucellin skin test. Thus, analysing the results of the 3 testing in parallel, 119 animals (94%) were positive to at least 1 of the performed tests. The results suggest that the use in parallel of the RB51 brucellin skin test with RB51-CFT may represent a reliable

  10. Confirmação de infecção por Brucella abortus em um rebanho bovino certificado livre em Minas Gerais: relato de caso

    OpenAIRE

    Soares Filho,P.M.; Wanderley,R.P.B.; Faria,G.C.; PENNA, A. G.; D.B.C.L. Ribeiro; Assis,R.A.; R.C. Leite; FONSECA JUNIOR, A. A.; A.C.C.L. Ribeiro

    2012-01-01

    Relata-se a ocorrência de um surto de brucelose em um rebanho de aproximadamente 1000 animais, livre da doença há 18 anos, certificado pelo Ministério da Agricultura, Pecuária e Abastecimento desde 2006. Dois animais reagiram aos testes sorológicos de diagnóstico por ocasião dos procedimentos de recertificação em 2008. Após o sacrifício deles, Brucella abortus, biovariedade 1, amostra não vacinal, foi isolada e identificada por meio de provas bioquímicas e de biologia molecular (PCR AMOS). A ...

  11. Antigenic, immunologic and genetic characterization of rough strains B. abortus RB51, B. melitensis B115 and B. melitensis B18.

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    Rosanna Adone

    Full Text Available The lipopolysaccharide (LPS is considered the major virulent factor in Brucella spp. Several genes have been identified involved in the synthesis of the three LPS components: lipid A, core and O-PS. Usually, Brucella strains devoid of O-PS (rough mutants are less virulent than the wild type and do not induce undesirable interfering antibodies. Such of them proved to be protective against brucellosis in mice. Because of these favorable features, rough strains have been considered potential brucellosis vaccines. In this study, we evaluated the antigenic, immunologic and genetic characteristics of rough strains B. abortus RB51, B. melitensis B115 and B. melitensis B18. RB51 derived from B. abortus 2308 virulent strain and B115 is a natural rough strain in which the O-PS is present in the cytoplasm. B18 is a rough rifampin-resistan mutant isolated in our laboratory. The surface antigenicity of RB51, B115 and B18 was evaluated by testing their ability to bind antibodies induced by rough or smooth Brucella strains. The antibody response induced by each strain was evaluated in rabbits. Twenty-one genes, involved in the LPS-synthesis, were sequenced and compared with the B. melitensis 16M strain. The results indicated that RB51, B115 and B18 have differences in antigenicity, immunologic and genetic properties. Particularly, in B115 a nonsense mutation was detected in wzm gene, which could explain the intracellular localization of O-PS in this strain. Complementation studies to evaluate the precise role of each mutation in affecting Brucella morphology and its virulence, could provide useful information for the assessment of new, attenuated vaccines for brucellosis.

  12. The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests.

    Science.gov (United States)

    Emmerzaal, A; de Wit, J J; Dijkstra, Th; Bakker, D; van Zijderveld, F G

    2002-02-01

    The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the programme to monitor the official Brucella-free status of bovine herds was primarily based on periodical testing of dairy herds with the milk ring test (MRT) and serological testing of all animals older than 1 year of age from non-dairy herds, using the micro-agglutination test (MAT) as screening test. In addition, serum samples of cattle that aborted were tested with the MAT. The high number of false positive reactions in both tests and the serum agglutination test (SAT) and complement fixation test (CFT) used for confirmation seemed to result in unnecessary blockade of herds, subsequent testing and slaughter of animals. For this reason, a validation study was performed in which three indirect enzyme-linked immunosorbent assays (ELISAs), the CFT and the SAT were compared using a panel of sera from brucellosis-free cattle, sera from experimentally infected cattle, and sera from cattle experimentally infected with bacteria which are known to induce cross-reactive antibodies (Pasteurella, Salmonella, Yersinia, and Escherichia). Moreover, four ELISAs and the MRT were compared using a panel of 1000 bulk milk samples from Brucella-free herds and 12 milk samples from Brucella abortus- infected cattle. It is concluded that the ELISA obtained from ID-Lelystad is the most suitable test to monitor the brucelosis free status of herds because it gives rise to fewer false-positive reactions than the SAT.

  13. Suplementación con selenio en vaquillas: Efecto sobre la respuesta inmune a las vacunas Brucella abortus cepa RB51 y toxoide tetánico Selenium suplementation in heifers: Effect on the immune response to Brucella abortus strain RB51 and tetanus toxoid vaccines

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    V Leyán

    2005-01-01

    Full Text Available El trabajo tiene por objetivo evaluar el efecto de una suplementación con selenio (Se sobre la respuesta inmune a las vacunas Toxoide tetánico y Brucella abortus cepa RB51 en vaquillas con un adecuado balance metabólico de selenio (GSH-Px >130 U/g Hb. Para ello se empelaron 32 vaquillas Friesian de 18 a 24 meses de edad, asignadas al azar a dos grupos de 16 animales; uno suplementado (Se-S el día 0 con una dosis de selenato de bario (1 mg Se/kg, s.c., permaneciendo el otro como control no suplementado (No-S. Todas las vaquillas fueron mantenidas durante 9 meses (abril a enero a pastoreo sobre una pradera naturalizada con un contenido de Se de 0,04 ppm/MS. Los animales fueron inmunizados con vacuna RB51 el día 60 y posteriormente con Toxoide tetánico los días 120 y 150. Muestras de sangre fueron obtenidas previo a la suplementación y cada 15 días hasta el término del experimento. El balance metabólico de selenio fue evaluado mediante la actividad sanguínea de Glutatión peroxidasa (GSH-Px. La respuesta inmune humoral se evaluó determinando los anticuerpos séricos específicos para ambos antígenos mediante ELISA y la respuesta inmune celular mediante pruebas de intradermorreacción a antígenos de Brucella abortus. La administración de Se aumentó (P 0,05 en ambos grupos experimentales, mientras que la respuesta celular a la vacuna RB51 fue menor (P The effect of selenium (Se supplementation on the immune response to tetanus toxoid and Brucella abortus strain RB51 vaccines was studied in heifers with a normal Se status (GSH-Px activity > 130 U/g Hb. Frisian heifers (n-32, 18 to 24 months old were randomly allocated into two groups of 16 animals each. Animals from one group were supplemented (Se-S with one dose of barium selenate (1 mg/Se/kg. s.c. on day 0; animals from the other group remained as a control without supplementation (No-S. The heifers grazed during 9 months (April to January a pasture that contained 0.04 ppm/DM of Se

  14. Caracterização epidemiológica e fatores de risco associados à infecção por Chlamydophila abortus em ovinos deslanados do semiárido brasileiro

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    Areano E.M. Farias

    2013-03-01

    Full Text Available O objetivo do presente trabalho foi determinar as prevalências de propriedades positivas e animais soropositivos para Chlamydophila abortus em ovinos deslanados da região semiárida do Nordeste do Brasil, bem como identificar fatores de risco. Foram colhidas amostras de sangue de 476 ovinos procedentes de 72 propriedades em 14 municípios na mesorregião do Sertão, Estado da Paraíba. Para o diagnóstico sorológico da infecção por Chlamydophila abortus foi utilizada a reação de fixação de complemento (RFC. Uma propriedade foi considerada positiva quando apresentou pelo menos um animal soropositivo. Das 72 propriedades usadas 38 (52,8% apresentaram pelo menos um animal soropositivo, e dos 476 animais 94 (19,7% foram soropositivos. Participação em exposições (odds ratio =4,31; IC 95% =1,80-10,35; p=0,011 foi identificada como fator de risco. Sugere-se que a infecção por Chlamydophila abortus encontra-se disseminada em ovinos da região, e baseando-se na análise de fatores de risco, recomenda-se o controle sanitário nas exposições de animais.

  15. Immunogenicity and Protective Response Induced by Recombinant Plasmids Based on the BAB1_0267 and BAB1_0270 Open Reading Frames of Brucella abortus 2308 in BALB/c Mice

    Science.gov (United States)

    Gómez, Leonardo A.; Alvarez, Francisco I.; Fernández, Pablo A.; Flores, Manuel R.; Molina, Raúl E.; Coloma, Roberto F.; Oñate, Angel A.

    2016-01-01

    Immunogenicity induced by recombinant plasmids based on the BAB1_0267 and BAB1_0270 open reading frames (ORFs) of Brucella abortus 2308 was evaluated. Bioinformatics analyses indicate that the BAB1_0267 and BAB1_0270 ORFs encode a protein with a SH3 domain and a Zn-dependent metalloproteinase, respectively. Both ORFs have important effects on intracellular survival and replication of B. abortus 2308, mediated via professional and non-professional phagocytic cells. Our results show that immunization with the recombinant plasmid based on the BAB1_0267 ORF significantly increases the production of IgG1, levels of IFN-γ and the lymphoproliferative response of splenocytes. However, BAB1_0267 did not provide significant levels of protection. The plasmid based on the BAB1_0270 significantly increased IgG2a production, levels of IFN-γ and TNF-α, and the lymphoproliferative response of splenocytes. These results demonstrate that immunization with the BAB1_0270 derived recombinant plasmid induce a Th1-type immune response, correlated with a heightened resistance to B. abortus 2308 infection in mice. It is concluded that the Th1-type immune response against bacterial Zn-dependent metalloproteinase induces a protective response in mice, and that pV270 recombinant plasmid is an effective candidate microbicide against brucellosis. PMID:27747197

  16. A serological study on Brucella abortus, caprine arthritis-encephalitis virus and Leptospira in dairy goats in Rio de Janeiro, Brazil.

    Science.gov (United States)

    Lilenbaum, Walter; de Souza, Guilherme Nunes; Ristow, Paula; Moreira, Madelayne Cortez; Fráguas, Suzana; Cardoso, Verônica da Silva; Oelemann, Walter Martin Roland

    2007-03-01

    In spite of the large number of goats found in several developing tropical countries, milk production remains unsatisfactory. The occurrence of infectious diseases, such as leptospirosis, brucellosis and caprine arthritis-encephalitis (CAE) may in part be responsible for sub-optimal production. In this study, 1000 serum samples were tested for leptospirosis, 953 for brucellosis and 562 for CAE. All tested flocks presented at least one seroreactive animal for leptospirosis and for CAE. Reactivity to leptospirosis was 11.1%, and serovar hardjo was the most frequently found. Anti-B. abortus agglutinins were found in 0.5% of the samples presented and 14.1% were seroreactive to CAE. Leptospirosis was considered to represent the major infectious problem in the studied goat flocks. The occurrence of infectious diseases in the tested flocks may represent an important factor contributing to the decreased productivity of the animals. These findings may be similar to those observed in other developing countries and require further study to define the relationship between seropositivity and reduced production.

  17. Improved performance of Brucella melitensis native hapten over Brucella abortus OPS tracer on goat antibody detection by the fluorescence polarization assay.

    Science.gov (United States)

    Ramírez-Pfeiffer, C; Díaz-Aparicio, E; Rodríguez-Padilla, C; Morales-Loredo, A; Alvarez-Ojeda, G; Gomez-Flores, R

    2008-06-15

    The current method for goat brucellosis diagnosis is based on the World Organization for Animal Health (OIE) using the screening card test (CT), with antigen at 8% (CT8) or 3% (CT3) of cell concentrations, and the confirmatory complement fixation test (CFT). However, these tests do not differentiate antibodies induced by vaccination from those derived from field infections by Brucella species or other bacterial agents; in places like Mexico, where the prevalence of brucellosis and the vaccination rates are high, there is a considerable percentage of false positive reactions that causes significant unnecessary slaughter of animals. Furthermore, results of the fluorescence polarization assay (FPA) using the Brucella abortus O-polysaccharide (OPS) tracer in goats are poorer than those with cattle. The present study was undertaken to investigate a tracer prepared from the native hapten (NH) of the Rev. 1 strain of Brucella melitensis to improve FPA performance on goat brucellosis diagnosis. Evaluation of 48 positive samples and 96 negative samples showed that the NH tracer was more accurate (pgoat sera samples selected by test series approved by the OIE (card test 3% and CFT). We demonstrated a new application for the NH lipopolysaccharide on detecting antibodies against Brucella using the FPA, which may yield faster results (minutes vs. 24-72h) than the immunodiagnosis assays frequently used in bovine brucellosis. In addition, NH tracer produces similar or better performance results than the conventional OPS tracer, using the FPA in goat sera samples.

  18. Distinct roles for hepcidin and interleukin-6 in the recovery from anemia in mice injected with heat-killed Brucella abortus.

    Science.gov (United States)

    Gardenghi, Sara; Renaud, Tom M; Meloni, Alessandra; Casu, Carla; Crielaard, Bart J; Bystrom, Laura M; Greenberg-Kushnir, Noa; Sasu, Barbra J; Cooke, Keegan S; Rivella, Stefano

    2014-02-20

    Anemia of inflammation (AI) is commonly observed in chronic inflammatory states and may hinder patient recovery and survival. Induction of hepcidin, mediated by interleukin 6, leads to iron-restricted erythropoiesis and anemia. Several translational studies have been directed at neutralizing hepcidin overexpression as a therapeutic strategy against AI. However, additional hepcidin-independent mechanisms contribute to AI, which are likely mediated by a direct effect of inflammatory cytokines on erythropoiesis. In this study, we used wild-type, hepcidin knockout (Hamp-KO) and interleukin 6 knockout (IL-6-KO) mice as models of AI. AI was induced with heat-killed Brucella abortus (BA). The distinct roles of iron metabolism and inflammation triggered by interleukin 6 and hepcidin were investigated. BA-treated wild-type mice showed increased expression of hepcidin and inflammatory cytokines, as well as transitory suppression of erythropoiesis and shortened red blood cell lifespan, all of which contributed to the severe anemia of these mice. In contrast, BA-treated Hamp-KO or IL-6-KO mice showed milder anemia and faster recovery compared with normal mice. Moreover, they exhibited different patterns in the development and resolution of anemia, supporting the notion that interleukin 6 and hepcidin play distinct roles in modulating erythropoiesis in AI.

  19. Confirmação de infecção por Brucella abortus em um rebanho bovino certificado livre em Minas Gerais: relato de caso

    Directory of Open Access Journals (Sweden)

    P.M. Soares Filho

    2012-10-01

    Full Text Available Relata-se a ocorrência de um surto de brucelose em um rebanho de aproximadamente 1000 animais, livre da doença há 18 anos, certificado pelo Ministério da Agricultura, Pecuária e Abastecimento desde 2006. Dois animais reagiram aos testes sorológicos de diagnóstico por ocasião dos procedimentos de recertificação em 2008. Após o sacrifício deles, Brucella abortus, biovariedade 1, amostra não vacinal, foi isolada e identificada por meio de provas bioquímicas e de biologia molecular (PCR AMOS. A origem do agente no rebanho é de difícil determinação. No entanto, a adoção de procedimentos preconizados pelo Programa Nacional de Controle e Erradicação da Brucelose permitiu evitar a disseminação da enfermidade. Ocorrências como essas, em que rebanhos livres foram infectados após anos sem a ocorrência de brucelose, nunca haviam sido relatadas no Brasil.

  20. Brucella melitensis Biovar 1 and Brucella abortus S19 Vaccine Strain Infections in Milkers Working at Cattle Farms in the Khartoum Area, Sudan.

    Directory of Open Access Journals (Sweden)

    Amira E F Osman

    Full Text Available Human brucellosis is a preventable zoonoses that may become persistent, causing, if left untreated, severe localized disease. Occupational exposure to infected animals or animal products and consumption of fresh contaminated dairy are main risk factors.One hundred farmworkers employed at two cattle farms one in Khartoum North and one in Omdurman were screened for the presence of specific antibodies and seropositive workers were invited to donate a blood sample for blood culture. Molecular typing was used to characterize Brucella isolates.Ten percent of farmworkers tested seropositive and while Brucella melitensis biovar 1 was isolated from the blood of three individuals, an isolate identical to the B. abortus S19 vaccine strain was isolated from a fourth person. All four bacteremic individuals were employed as milkers and did not have obvious disease.The isolation of the highly infectious pathogen B. melitensis from seropositive workers is consistent with the notion that the pathogen may persist in the blood without causing overt disease. While vaccination with strain S19 is essential for the control of bovine brucellosis the vaccine strain may be transmitted to the human population and protective measures remain important to prevent exposure also in view of the presence of B. melitensis. To create awareness for this potentially severe disease more information on the prevalence of the pathogen in different risk groups and in livestock in the Sudan is needed.

  1. A serological prevalence survey of Brucella abortus in cattle of rural communities in the province of KwaZulu-Natal, South Africa : article

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    U.W. Hesterberg

    2008-05-01

    Full Text Available A serological survey of Brucella abortus in cattle originating from communal grazing areas of Kwa Zulu Natal was carried out between March 2001 and December 2003. The survey was designed as a 2-stage survey, considering the diptank as the primary sampling unit. In total 46 025 animals from 446 diptanks of 33 magisterial districts were sampled and tested using the Rose Bengal test and Complement Fixation Test. The apparent prevalence at district level was adjusted for clustering, diagnostic test sensitivity and specificity, and mapped using ArcView version 3.3. The prevalence of brucellosis in communal grazing areas of Kwa-Zulu Natal was found to be 1.45 % (0.84-2.21 % and varied from 0 to 15.6% between magisterial districts. In 19 of the 33 magisterial districts no serological reactors were observed. A large variation in prevalence was found within diptank areas. Brucellosis was found to be most prevalent in the northeastern area of the province. The findings of the survey are discussed.

  2. Immunization of BALB/c mice with Brucella abortus 2308ΔwbkA confers protection against wild-type infection.

    Science.gov (United States)

    Li, Zhi-qiang; Gui, Dan; Sun, Zhi-hua; Zhang, Jun-bo; Zhang, Wen-zhi; Zhang, Hui; Guo, Fei; Chen, Chuang-fu

    2015-01-01

    Brucellosis is a zoonotic disease that causes animal and human diseases. Vaccination is a major measure for prevention of brucellosis, but it is currently not possible to distinguish vaccinated animals from those that have been naturally infected. Therefore, in this study, we constructed the Brucella (B.) abortus 2380 wbkA mutant (2308ΔwbkA) and evaluated its virulence. The survival of 2308ΔwbkA was attenuated in murine macrophage (RAW 264.7) and BALB/c mice, and it induced high protective immunity in mice. The wbkA mutant elicited an anti-Brucella-specific immunoglobulin G response and induced the secretion of gamma interferon. Antibodies to 2308ΔwbkA could be detected in sera from mice, implying the potential for use of this protein as a diagnostic antigen. The WbkA antigen would allow serological differentiation between infected and vaccinated animals. These results suggest that 2308ΔwbkA is a potential attenuated vaccine against 16M. This vaccine will be further evaluated in sheep.

  3. A new cis-encoded sRNA, BsrH, regulating the expression of hemH gene in Brucella abortus 2308.

    Science.gov (United States)

    Peng, Xiaowei; Dong, Hao; Wu, Qingmin

    2015-01-01

    A total of 129 sRNA candidates were identified in Brucella abortus 2308 in our previous work, and one candidate with potential to regulate expression of hemH gene was further analyzed in this study. We found that the novel sRNA can inhibit the expression of hemH and called it BsrH (Brucella sRNA regulating HemH). The expression level of BsrH was tested in four different stress conditions. A significant upregulation was detected during the growth in acidic and Brucella minimal media, as well as in the presence of hydroxyl peroxide, while iron deficiency caused the opposite effect. As expected, BsrH strongly affected the survival ratio of the Brucella cells under iron-limitation conditions, though overexpression of BsrH did not affect Brucella virulence. Thus, we conclude that BsrH plays a regulatory role in bacterial heme biosynthesis and can be considered as the first Brucella sRNA involved in stress responses.

  4. 快速检测牛羊猪种布鲁杆菌多重PCR的建立%Differentiation of Brucella abortus , Brucella melitensis , and Brucella suis by multiple primers PCR

    Institute of Scientific and Technical Information of China (English)

    刘锴; 王兴龙; 马鸣萧; 翟丽鹃

    2009-01-01

    Objective To establish a method for rapidly identifying Brucella abortus, Brucella melitensis and Brucella suis by multiple primers PCR. Methods According to Brncella abortus, Brucella melitensis and Brucella suis IS711 insertion sequences, a public primer and three specific primers(544A, 16M, 1330S) were designed to set up multiplex PCR detection method. Yersinia O : 9, Escherichia coli O157 : HT, Salmonella typhimurium 47729 were selected to undergo multiple PCR reactions to detect the specificity. The sensitivity of multiple primers PCR of Brucella abortus was detected using multiple proportion dilution method. Results The amplified fragment size of Brucella abortus was 485 bp, that of Brucella melitensis 731 bp, and that of Brucella suis 248 bp, but PCR for the DNA of Yersinia O : 9, Escherichia coli O157 : H7, Salmonella typhimurium 47729 was negative. A sensitivity of the multiple primers PCR with Brucella abortus DNA using multiple proportional dilution quantitative method was 0.0967 pg. Conclusions Multiple PCR amplification method for rapidly detecting Brucella abortus, Brucella melitensis and Brucella suis has been successfully established, resulting in good specificity and sensitivity.%目的 建立一种能同时快速检测并能鉴别牛、羊、猪种布鲁杆菌的多重PCR方法.方法 根据IS711插入序列设计1条公共引物和3条牛、羊、猪种布鲁杆菌(544A、16M、1330S)特有序列引物,进行多重PCR反应;选择耶尔森菌O:9、大肠埃希菌O157:H7、鼠伤寒沙门菌47729进行多重PCR反应的特异性检测;倍比稀释定量法观察牛种布鲁杆菌多重PCR反应的敏感性.结果 牛、羊、猪种布鲁杆菌多重PCR反应扩增片段产物长度分别为485、731、248 bp;耶尔森菌O:9、大肠埃希菌O157:H7、鼠伤寒沙门菌47729加入布鲁杆菌中进行多重PCR反应.扩增结果呈阴性;牛种布鲁杆菌多重PCR反应敏感性为0.0967 Pg.结论 成功建立快速检测牛、羊、猪种布鲁

  5. Chlamydophilose abortive ovine : études à propos d'une suspicion de résistance de Chlamydophila abortus au vaccin vivant thermosensible dans des élevages ovins laitiers du rayon de Roquefort

    OpenAIRE

    Uhart, Maia

    2009-01-01

    Des cas d'avortements imputés à Chlamydophila abortus ont été décrits dans des élevages de brebis du rayon Roquefort (France, 12) alors que ces élevages pratiquaient la vaccination contre la chlamydophilose avec un vaccin vivant depuis plusieurs années. De brefs rappels bibliographiques concernant la chlamydophilose et la vaccination contre cette maladie sont présentés en première partie. Une enquête menée auprès de neuf éleveurs est ensuite exposée afin d'expliquer la persistance d'avortemen...

  6. Producción de interferón gamma en cultivos de sangre completa en respuesta a antígenos de Brucella abortus en bovinos vacunados con RB51

    OpenAIRE

    Marisela Leal Hernández; Laura Jaramillo-Meza; Laura Hernández-Andrade; Rafael Pérez-González; Efrén Díaz-Aparicio; Francisco Suárez Güemes

    2007-01-01

    Con el propósito de evaluar la capacidad inductora de la producción de interferón gama de diferentes preparaciones proteínicas, y de los lipopolisacáridos liso y rugoso de Brucella abortus en cultivos de sangre completa de bovinos inmunizados con RB51, se seleccionaron animales vacunados y revacunados de un hato con elevada prevalencia de la enfermedad; igualmente, un grupo de animales seronegativos de un hato control fue considerado para valorar la utilidad del ensayo. Cultivos de sangre com...

  7. Imunodifusão em gel de ágar com polissacarídeos de membrana de Brucella abortus 1119-3 no diagnóstico da brucelose bovina

    Directory of Open Access Journals (Sweden)

    Megid J.

    1999-01-01

    Full Text Available Comparou-se a prova de imunodifusão em gel de ágar (IDGA, utilizando extrato polissacarídico (POLI O, obtido da amostra de B. abortus 1119-3, com os testes de soroaglutinação rápida em placa, de soroaglutinação lenta em tubos, de antígeno acidificado e de 2-mercaptoetanol para o diagnóstico da brucelose bovina. O IDGA mostrou alta especificidade, porém sensibilidade inferior aos métodos convencionais.

  8. Prevalência e fatores de risco associados à infecção por Chlamydophila abortus em granjas suinícolas tecnificadas no Estado de Alagoas, Brasil

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    Rômulo M. B. Valença

    2011-01-01

    Full Text Available Objetivou-se com este estudo calcular a prevalência e identificar os fatores de risco associados à infecção por Chlamydophila abortus em suínos criados em granjas tecnificadas no Estado de Alagoas, Brasil. Para compor a amostra do estudo foram utilizados 342 suínos, sendo 312 matrizes e 30 varrões oriundos de sete granjas de ciclo completo e distribuídas em cinco municípios do Estado de Alagoas. O diagnóstico sorológico da infecção por C. abortus foi realizado através da microtécnica de Fixação do Complemento (RFC. A análise dos fatores de risco foi realizada por meio da aplicação de questionários investigativos, constituídos por perguntas objetivas referentes ao criador, às características gerais da propriedade, ao manejo produtivo, reprodutivo e sanitário. Observou-se prevalência de 10,5% (36/342 de suínos soropositivos para a infecção por C. abortus, com 85,8% das granjas analisadas com animais positivos. As variáveis que demonstraram associação significativa foram: utilização de bebedouros comuns para jovens e adultos (p=0,024; OR=10,83; IC=1,36-86,03 e método de cobertura de monta natural associada à inseminação artificial (p=0,05; OR=7,62; IC=1,00-58,31. Relata-se a primeira ocorrência de anticorpos anti-C. abortus em suínos no Brasil. Fatores como a introdução de reprodutores nos plantéis e a forma de fornecimento de água foram evidenciados como facilitadores da infecção das matrizes neste estudo. Dessa forma, medidas de controle da infecção devem ser enfocadas nesse aspecto para evitar a disseminação do agente nas granjas suinícolas e em outros plantéis da região.

  9. First evaluation of an influenza viral vector based Brucella abortus vaccine in sheep and goats: Assessment of safety, immunogenicity and protective efficacy against Brucella melitensis infection.

    Science.gov (United States)

    Tabynov, Kaissar; Yespembetov, Bolat; Matikhan, Nurali; Ryskeldinova, Sholpan; Zinina, Nadezhda; Kydyrbayev, Zhailaubay; Assanzhanova, Nurika; Tabynov, Kairat; Renukaradhya, Gourapura J; Mukhitdinova, Gulnara; Sansyzbay, Abylai

    2016-12-25

    Previously we developed and evaluated a candidate influenza viral vector based Brucella abortus vaccine (Flu-BA) administered with a potent adjuvant Montanide Gel01 in cattle, which was found safe and highly effective. This study was aimed to establish a proof-of-concept of the efficacy of Flu-BA vaccine formulation in sheep and goats. We vaccinated sheep and goats with Flu-BA vaccine and as a positive control vaccinated a group of animals with a commercial B. melitensis Rev.1 vaccine. Clinically, both Flu-BA and Rev.1 vaccines were found safe. Serological analysis showed the animals received Flu-BA vaccine did not induce antibody response against Brucella Omp16 and L7/L12 proteins during the period of our study (56days post-initial vaccination, PIV). But observed significant antigen-specific T cell response indicated by increased lymphocyte stimulation index and enhanced secretion of IFN-γ at day 56 PIV in Flu-BA group. The Flu-BA vaccinated animals completely protected 57.1% of sheep and 42.9% of goats against B. melitensis 16M challenge. The severity of brucellosis in terms of infection index and colonization of Brucella in tissues was significantly lower in the Flu-BA group compared to negative control animals group. Nevertheless, positive control commercial Rev.1 vaccine provided strong antigen-specific T cell immunity and protection against B. melitensis 16M infection. We conclude that the Flu-BA vaccine induces a significant antigen-specific T-cell response and provides complete protection in approximately 50% of sheep and goats against B. melitensis 16M infection. Further investigations are needed to improve the efficacy of Flu-BA and explore its practical application in small ruminants.

  10. Brucella abortus surveillance of cattle in Fiji, Papua New Guinea, Vanuatu, the Solomon Islands and a case for active disease surveillance as a training tool.

    Science.gov (United States)

    Tukana, Andrew; Hedlefs, Robert; Gummow, Bruce

    2016-10-01

    There have been no surveys of the cattle population for brucellosis in the Pacific Island Countries and Territories (PICTs) for more than 15 years. This study used disease surveillance as a capacity building training tool and to examine some of the constraints that impede surveillance in PICTs. The study also developed and implemented a series of surveys for detecting antibodies to B. abortus in cattle in Fiji, Papua New Guinea, Vanuatu and the Solomon Islands contributing to OIE requirements. The findings indicated lack of funds, lack of technical capacity, shortage of veterinarians, high turnover of in-country officials and lack of awareness on the impacts of animal diseases on public health that were constraining active disease surveillance. During the development and implementation of the surveys, constraints highlighted were outdated census data on farm numbers and cattle population, lack of funds for mobilisation of officials to carry out the surveys, lack of equipment for collecting and processing samples, lack of staff knowledge on blood sampling, geographical difficulties and security in accessing farms. Some of the reasons why these were constraints are discussed with likely solutions presented. The detection surveys had the objectives of building capacity for the country officials and demonstrating freedom from brucellosis in cattle for PNG, Vanuatu and the Solomon Islands. PNG, Vanuatu and the Solomon Islands all demonstrated freedom from bovine brucellosis in the areas surveyed using the indirect ELISA test. Fiji had an outbreak of brucellosis, and the objective was to determine its distribution and prevalence on untested farms. The Muaniweni district surveyed during the training had a 95 % confidence interval for true prevalence between 1.66 and 5.45 %. The study showed that active disease surveillance could be used as a tool for training officials thus, improves surveillance capacity in resource poor countries.

  11. Overproduced Brucella abortus PdhS-mCherry forms soluble aggregates in Escherichia coli, partially associating with mobile foci of IbpA-YFP

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    Matroule Jean-Yves

    2010-09-01

    Full Text Available Abstract Background When heterologous recombinant proteins are produced in Escherichia coli, they often precipitate to form insoluble aggregates of unfolded polypeptides called inclusion bodies. These structures are associated with chaperones like IbpA. However, there are reported cases of "non-classical" inclusion bodies in which proteins are soluble, folded and active. Results We report that the Brucella abortus PdhS histidine kinase fused to the mCherry fluorescent protein forms intermediate aggregates resembling "non-classical" inclusion bodies when overproduced in E. coli, before forming "classical" inclusion bodies. The intermediate aggregates of PdhS-mCherry are characterized by the solubility of PdhS-mCherry, its ability to specifically recruit known partners fused to YFP, suggesting that PdhS is folded in these conditions, and the quick elimination (in less than 10 min of these structures when bacterial cells are placed on fresh rich medium. Moreover, soluble PdhS-mCherry foci do not systematically colocalize with IpbA-YFP, a marker of inclusion bodies. Instead, time-lapse experiments show that IbpA-YFP exhibits rapid pole-to-pole shuttling, until it partially colocalizes with PdhS-mCherry aggregates. Conclusion The data reported here suggest that, in E. coli, recombinant proteins like PdhS-mCherry may transit through a soluble and folded state, resembling previously reported "non-classical" inclusion bodies, before forming "classical" inclusion bodies. The dynamic localization of IbpA-YFP foci suggests that the IbpA chaperone could scan the E. coli cell to find its substrates.

  12. TLR2 and TLR4 signaling pathways are required for recombinant Brucella abortus BCSP31-induced cytokine production, functional upregulation of mouse macrophages, and the Th1 immune response in vivo and in vitro.

    Science.gov (United States)

    Li, Jia-Yun; Liu, Yuan; Gao, Xiao-Xue; Gao, Xiang; Cai, Hong

    2014-09-01

    Brucella abortus is a zoonotic Gram-negative pathogen that causes brucelosis in ruminants and humans. Toll-like receptors (TLRs) recognize Brucella abortus and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. In this study, we focused on recombinant Brucella cell-surface protein 31 (rBCSP31) to determine its effects on mouse macrophages. Our results demonstrated that rBCSP31 induced TNF-α, IL-6 and IL-12p40 production, which depended on the activation of mitogen-activated protein kinases (MAPKs) by stimulating the rapid phosphorylation of p38 and JNK and the activation of transcription factor NF-κB in macrophages. In addition, continuous exposure (>24 h) of RAW264.7 cells to rBCSP31 significantly enhanced IFN-γ-induced expression of MHC-II and the ability to present rBCSP31 peptide to CD4(+) T cells. Furthermore, we found that rBCSP31 could interact with both TLR2 and TLR4. The rBCSP31-induced cytokine production by macrophages from TLR2(-/-) and TLR4(-/-) mice was lower than that from C57BL/6 macrophages, and the activation of NF-κB and MAPKs was attenuated in macrophages from TLR2(-/-) and TLR4(-/-) mice. In addition, CD4(+) T cells from C57BL/6 mice immunized with rBCSP31 produced higher levels of IFN-γ and IL-2 compared with CD4(+) T cells from TLR2(-/-) and TLR4(-/-) mice. Macrophages from immunized C57BL/6 mice produced higher levels of IL-12p40 than those from TLR2(-/-) and TLR4(-/-) mice. Furthermore, immunization with rBCSP31 provided better protection in C57BL/6 mice than in TLR2(-/-) and TLR4(-/-) mice after B. abortus 2308 challenge. These results indicate that rBCSP31 is a TLR2 and TLR4 agonist that induces cytokine production, upregulates macrophage function and induces the Th1 immune response.

  13. 集约化养猪场嗜流产衣原体病流行状况的初步调查%Preliminary Investigation of Chlamydophila abortus in Intensive Pig Farms

    Institute of Scientific and Technical Information of China (English)

    田德雨; 刘哲翔; 李靖; 杨君敬; 信先; 汪明; 何诚

    2012-01-01

    本试验对北京郊区5个规模化猪场断奶仔猪、育成猪、育肥猪和怀孕母猪采集血清共507份,同时采集密切接触饲养人员、仔猪、育肥猪的喉头拭子、母猪阴道拭子及公猪的精液共183份.分别使用间接血凝诊断试剂盒、法国ELISA试剂盒检测抗体;利用直接荧光染色法检测抗原,包括166份猪喉头拭子、阴道拭子和精液以及17份饲养人员喉头拭子.结果发现间接血凝诊断试剂盒、ELISA试剂盒抗体阳性率分别为4.14%和2.17%;抗原平均阳性率为14.8%,其中精液阳性率达到37.5%,母猪阴道拭子阳性率为27.5%,饲养人员阳性率为23.5%.本试验证实北京郊区猪嗜流产衣原体在种公猪、母猪群和饲养员呈高流行趋势.同时研究结果显示,5个被调查的规模化猪场嗜流产衣原体抗体阳性率显著低于有报道的其他省市,这与间接血凝诊断试剂盒检测结果高于ELISA试剂有关.针对发现的问题,必须采取有效措施控制种公猪精液污染衣原体现象,阻断向母猪和饲养人员传播,以降低人兽共患病发生的风险.%In order to evaluate prevalence of swine Chlamydophila abortus in Beijing intensive pig farms, 507 blood samples were detected for antibodies using indirectly hemorrhage assay (IHA) and ELISA while 183 clinical samples were determined for positive antigens by immunofluorescence test kits(IF), including 166 samples from pig swabs and pig sperms and 17 throat swabs from pig farmers. The positive antibody against Chlamydophila abortus was 4. 14% (IHA) and 2. 17% (ELISA). Respectively while the average positive antigen was average 14. 8% (IF). Moreover, the higher antigen positivity was found in the boar sperm samples, sow virginal swabs and in throat swabs from pig farmers, which accounted for 37. 5%, 27. 5% and 23. 5% , respectively in the survey. On contrary to the previous reports, the positive antibody in Beijing intensive pig farms was found

  14. 改进试管凝集试验法检测布鲁氏菌病抗体%Detection of Anti-Brucella abortus Sera Agglutination Titers with Modified Tube Agglutination Test

    Institute of Scientific and Technical Information of China (English)

    李翠; 关孚时; 戴志红; 蒋卉; 温芳; 陆连寿; 王在时

    2012-01-01

    In order to prove the use value of the modified tube agglutination test,six anti-Brucella abortus sera were detected with MSAT and SAT referred to the international standard.The results showed that the sera titers were almost consistent detected with SAT and MSAT and the advantages of MSAT could be list as follows: The dilution method was simpler and more accurate.We could save time and effort,and save the materials such as serum,antigen etc.The result was easier to determine and with good reproducibility.%为证实改进的试管凝集试验(SAT)的使用价值,分别用微量试管凝集试验(MSAT)和SAT以国际标准血清为参比检测6份牛布鲁氏菌病阳性血清的抗体效价。结果表明:MSAT和SAT试验结果一致,而且MSAT具有稀释方法更简便、精确,省时省力,可节约血清、抗原等试验原材料,试验结果容易判定,重现性良好等优越性。

  15. Ocorrência de anticorpos anti-Brucella abortus e anti-Brucella canis em cães rurais e urbanos do Município de Monte Negro, Rondônia, Brasil

    Directory of Open Access Journals (Sweden)

    Aguiar Daniel Moura de

    2005-01-01

    Full Text Available Foram avaliados 304 cães de ambiente rural e urbano do município de Monte Negro, Rondônia, através do Antígeno Acidificado Tamponado (AAT, Soroaglutinação Lenta em Tubos (SAL e 2-Mercaptoetanol (2-ME para a pesquisa de anticorpos anti-Brucella abortus e da Imunodifusão em gel de ágar (IDGA e Imunodifusão em gel de ágar com soro tratado com 2-Mercaptoetanol (IDGA-ME para Brucella canis. Foram consideradas positivas as amostras reagentes nas provas confirmatórias do 2-ME e IDGA-ME. Verificaram-se 56 (18,4% animais reagentes ao AAT e 12 (4,0% reagentes a SAL. Apenas um cão (0,3% foi considerado positivo, confirmado pela prova do 2-ME. Foram observadas 11 (3,6% reações á IDGA, porém não houve confirmação na prova do IDGA-ME. Ressalta-se a baixa ocorrência de cães positivos ao 2-ME e a ausência de animais reagentes á IDGA-ME.

  16. S-SAD phasing of monoclinic histidine kinase from Brucella abortus combining data from multiple crystals and orientations: an example of data-collection strategy and a posteriori analysis of different data combinations.

    Science.gov (United States)

    Klinke, Sebastián; Foos, Nicolas; Rinaldi, Jimena J; Paris, Gastón; Goldbaum, Fernando A; Legrand, Pierre; Guimarães, Beatriz G; Thompson, Andrew

    2015-07-01

    The histidine kinase (HK) domain belonging to the light-oxygen-voltage histidine kinase (LOV-HK) from Brucella abortus is a member of the HWE family, for which no structural information is available, and has low sequence identity (20%) to the closest HK present in the PDB. The `off-edge' S-SAD method in macromolecular X-ray crystallography was used to solve the structure of the HK domain from LOV-HK at low resolution from crystals in a low-symmetry space group (P21) and with four copies in the asymmetric unit (∼108 kDa). Data were collected both from multiple crystals (diffraction limit varying from 2.90 to 3.25 Å) and from multiple orientations of the same crystal, using the κ-geometry goniostat on SOLEIL beamline PROXIMA 1, to obtain `true redundancy'. Data from three different crystals were combined for structure determination. An optimized HK construct bearing a shorter cloning artifact yielded crystals that diffracted X-rays to 2.51 Å resolution and that were used for final refinement of the model. Moreover, a thorough a posteriori analysis using several different combinations of data sets allowed us to investigate the impact of the data-collection strategy on the success of the structure determination.

  17. Generation and characterization of β1,2-gluco-oligosaccharide probes from Brucella abortus cyclic β-glucan and their recognition by C-type lectins of the immune system

    Science.gov (United States)

    Zhang, Hongtao; Palma, Angelina S; Zhang, Yibing; Childs, Robert A; Liu, Yan; Mitchell, Daniel A; Guidolin, Leticia S; Weigel, Wilfried; Mulloy, Barbara; Ciocchini, Andrés E; Feizi, Ten; Chai, Wengang

    2016-01-01

    The β1,2-glucans produced by bacteria are important in invasion, survival and immunomodulation in infected hosts be they mammals or plants. However, there has been a lack of information on proteins which recognize these molecules. This is partly due to the extremely limited availability of the sequence-defined oligosaccharides and derived probes for use in the study of their interactions. Here we have used the cyclic β1,2-glucan (CβG) of the bacterial pathogen Brucella abortus, after removal of succinyl side chains, to prepare linearized oligosaccharides which were used to generate microarrays. We describe optimized conditions for partial depolymerization of the cyclic glucan by acid hydrolysis and conversion of the β1,2-gluco-oligosaccharides, with degrees of polymerization 2–13, to neoglycolipids for the purpose of generating microarrays. By microarray analyses, we show that the C-type lectin receptor DC-SIGNR, like the closely related DC-SIGN we investigated earlier, binds to the β1,2-gluco-oligosaccharides, as does the soluble immune effector serum mannose-binding protein. Exploratory studies with DC-SIGN are suggestive of the recognition also of the intact CβG by this receptor. These findings open the way to unravelling mechanisms of immunomodulation mediated by β1,2-glucans in mammalian systems. PMID:27053576

  18. Brucellosis outbreak in a rural endemic region of Mexico - a comprehensive investigation.

    Science.gov (United States)

    Morales-Garcia, Maria Rosario Morales-Garcia; Lopez-Mendez, Jaime; Pless, Reynaldo; Garcia-Morales, Emilio; Kosanke, Hannah; Hernandez-Castro, Rigoberto; Bedi, Jasbir; Lopez-Merino, Ahide; Velazquez-Guadarram, Norma; Jimenez-Rojas, Leticia; Rontreras-Rodriguez, Araceli

    2015-01-01

    Brucellosis is a worldwide zoonotic disease. Generally, humans can be infected by either the consumption of raw milk and fresh cheeses made from unpasteurised milk or by contact with infected animals, mainly in endemic regions. In this study, we investigated a brucellosis outbreak in State of Guanajuato, an endemic region of Mexico. Microbiological culture of human blood, raw milk from cows and goats, and fresh cheeses was performed to isolate Brucella. Identification of the bacteria was done by bacteriological procedures and by multiplex Bruce-ladder polymerase chain reaction (PCR). Brucella melitensis was isolated from patients, infected goats, and fresh goat cheeses; while Brucella abortus was isolated from cows. All patients had eaten fresh cheese, but no occupational exposure to animals was reported. The results of molecular typing did not show any Brucella vaccine strains. The isolation, identification, and molecular characterisation of Brucella spp. in both human brucellosis cases and infected animals are very important to identify the source of infection and to take control measures in endemic regions.

  19. PLGA (85:15) nanoparticle based delivery of rL7/L12 ribosomal protein in mice protects against Brucella abortus 544 infection: A promising alternate to traditional adjuvants.

    Science.gov (United States)

    Singh, Damini; Somani, Vikas Kumar; Aggarwal, Somya; Bhatnagar, Rakesh

    2015-12-01

    There is a compelling need for the development of suitable adjuvants for human use to enhance the efficacy of the upcoming vaccines for the prevention of life threatening infections. In the current study, we have tried to explore the immunogenic potential of nanoparticles (NPs) made of PLGA (poly lactic-co-glycolic acid), a biodegradable and biocompatible polymer approved by FDA for human use after entrapping rL7/L12 protein, an immunodominant antigen of Brucella. Adjuvant properties were exhibited by the formulation as it elicited high IgG antibody titers just after first immunization which increased significantly after the booster administration. A good elicitation of the Th1 cytokines especially IFN-γ was recorded. Amongst the IgG antibody subclasses, IgG1 remained the predominant subclass to be elicited in mice serum after immunization; however IgG1/2a ratio showed a mixed profile of Th1/Th2 response. Lymphocyte proliferation assay as a marker of amplification in cellular immunity demonstrated that the splenocytes of the immunized mice had a high proliferation index with reference to the control, revealing that L7/L12 entrapping PLGA nanoparticles are potent inducer of inflammatory cell response indispensable to combat Brucella infection. Enumeration of splenic CFU after 14 days of infection with Brucella abortus 544 showed a significant reduction in log CFU of splenic bacteria in the vaccinated mice as compared to the control group. Therefore it is evident that PLGA nano formulations delivering the entrapped vaccine candidate in mice elicit specific humoral as well as cellular responses specific to the entrapped Brucella antigen. So there is much promise in this approach and this work by highlighting the adjuvant properties of the PLGA nanospheres will accelerate the development of improved vaccines safe for human as well as veterinary use.

  20. Regional Competition in Maghreb Region

    Institute of Scientific and Technical Information of China (English)

    EL AMOURI ALLAL

    2016-01-01

    Great powers focus on cooperation instead of competition in their regions in order to maintain stability and develop the economy of their regions and the world in general. The United States for example created the North American Free Trade Agreement (NAFT

  1. Multiplex fluorescence quantitative PCR detection on Brucella spp.,Brucella abortus, and Brucella melitensis%多重荧光定量PCR方法鉴定布鲁氏菌属及牛羊种布鲁氏菌研究

    Institute of Scientific and Technical Information of China (English)

    刘志国; 罗成旺; 张利; 姜海; 赵鸿雁; 朴东日; 田国忠; 崔步云

    2012-01-01

    verified by amplification of 97 local strains. Results from multiplex TaqMan real-time PCR detection of Brucella showed that only Brucella spp. , Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) had respective fluorescence signal for each of them; while there was no fluorescent signal for pathogens with closer homology. According to standard curve, the lower limit of detection for both single and multiplex fluorescent PCR was 102copy/μL (13-24 fg/μL) , 100 times higher than that of conventional PCR. It ' s suggested that with advantages of higher sensitivity and fast and easy operation, the new method could be used to conduct rapid identification of Brucella spp. , B. abortus and B. melitensis in a single test.

  2. Regional odontodysplasia.

    Science.gov (United States)

    Mehta, D N; Bailoor, D; Patel, B

    2011-01-01

    Regional odontodysplasia is an unusual developmental anomaly in which ectodermal and mesodermal tooth components are affected. We present a rare case of a developmental anomaly called regional odontodysplasia or 'ghost teeth' in a 12-year-old Indian girl. The anomaly affected right maxillary permanent teeth. The mandibular teeth were unaffected. The clinical, radiographic and histological features are reviewed. The management of affected patients is discussed.

  3. Regional odontodysplasia

    Directory of Open Access Journals (Sweden)

    D N Mehta

    2011-01-01

    Full Text Available Regional odontodysplasia is an unusual developmental anomaly in which ectodermal and mesodermal tooth components are affected. We present a rare case of a developmental anomaly called regional odontodysplasia or ′ghost teeth′ in a 12-year-old Indian girl. The anomaly affected right maxillary permanent teeth. The mandibular teeth were unaffected. The clinical, radiographic and histological features are reviewed. The management of affected patients is discussed.

  4. 地理信息系统在青海省布鲁杆菌病菌种空间分布中的应用研究%Application of a geographic information system on spatial distribution of Brucella abortus species in Qinghai Province

    Institute of Scientific and Technical Information of China (English)

    秦豫民; 徐立青; 马俊英; 杨旭欣; 田广; 胡桂英; 薛红梅; 杨宁海; 马丽

    2013-01-01

    Objective To establish a geographic information system for Brucella abortus species and study the spatial distribution of Brucella abortus species and strains in Qinghai Province.Methods We got the strains'information from Brucellosis Prevention and Control Department,Qinghai Institute for Endemic Diseases Prevention and Control.Through the geographical information technology software,we established the geographic information system database for Brucella abortus species in Qinghai Province and system fast query function.Time and space distribution of Brucella abortus species and strains was retrieved by the system fast query function and the layer set.Results Through the system fast query function,we got 113 strains' time distribution information.Through the layer set of time and space distribution for spatial distribution,we got 113 strains' and 3 species' spatial distribution information.One hundred and thirteen strains distributed in 21 counties(towns,cities) of Qinghai Province,most in Dulan County (21 strains),Qilian County (16 strains),Dachaidan Town (15 strains),Xinghai County(13 strains) and Gonghe County(10 strains).One hundred and three Brucella melitensis strains distributed in 17 counties(towns,cities) of Qinghai Province,most in Dulan County(20 strains),Qilian County(16 strains),Dachaidan Town (15 strains),Xinghai County (13 strains) and Gonghe County (10 strains); 8 Brucella abortus strains distributed in 6 counties(cities) ; 2 Brucella suis strains distributed in 2 counties.Conclusions We have successfully established a geographic information system for Brucella abortus species and strains in Qinghai Province.Through the layer set for spatial distribution,we could intuitively observe the spatial distribution of Brucella abortus species and strains in Qinghai Province.%目的 建立布鲁杆菌菌种地理信息系统,了解青海省布鲁杆菌病菌株与菌种的空间分布.方法 113株布鲁杆菌菌株资料来源于青海省地方病预防控

  5. Regional odontodysplasia

    Directory of Open Access Journals (Sweden)

    Thimma Reddy B

    2010-01-01

    Full Text Available Regional odontodysplasia (ROD is a rare developmental anomaly involving both mesodermal and ectodermal components in a group of contiguous teeth. It affects the primary and permanent dentitions in the maxilla and the mandible or both, however, the maxilla is frequently involved. Although the etiology of the ROD is uncertain, it has been suggested that numerous other factors play a role. The treatment plan should be based on the degree of involvement as well as the functional and esthetic needs in each case. This article reports the case of a 5-year-old boy presenting a rare anomaly on the right side of the maxillary arch. The treatment performed was rehabilitation with temporary partial acrylic denture and periodic checkups. The extraction was followed by rehabilitation with dental implants. The main aim of this article is to provide valuable information to pediatric dentists about the review and treatment alternatives for ROD.

  6. Desenvolvimento regional: a diversidade regional como potencialidade

    Directory of Open Access Journals (Sweden)

    José Odim Degrandi

    2013-04-01

    Full Text Available Throughout the second half of the twentieth century the debate on regional development in Brazil received special attention, especially among economists and geographers. Two main approaches stand out in the debate: one that understands the promotion of regional development from the reduction / elimination of regional inequalities, and the other proposes the understanding of regional diversity and potentiality for developing regions. The first was and still remains hegemonic, being evidenced both in scientific papers and in documents guiding public policies in the area. The latter is evidenced most clearly from the 1990s on, a period when the concept of region is resumed under a new approach. This paper aims to clarify the meaning of each of these concepts in order to contribute to the qualification of the discussion on regional development.

  7. Desenvolvimento regional: a diversidade regional como potencialidade

    Directory of Open Access Journals (Sweden)

    Virginia Elisabeta Etges

    2013-03-01

    Full Text Available Ao longo da segunda metade do século XX o debate sobre desenvolvimento regional no Brasil recebeu atenção especial, principalmente, entre economistas e geógrafos. Dois enfoques principais se destacam no debate: um, que entende a promoção do desenvolvimento regional a partir da redução/eliminação das desigualdades regionais; outro, que propõe a compreensão da diversidade regional como potencialidade para o desenvolvimento de regiões. O primeiro foi e continua sendo hegemônico, evidenciado tanto em trabalhos científicos quanto em documentos norteadores das políticas públicas na área. Já o segundo evidencia-se mais claramente a partir da década de 1990, período em que o conceito de região é retomado sob um novo enfoque. Neste trabalho pretende-se explicitar o significado de cada uma dessas concepções, no intuito de contribuir para a qualificação da discussão sobre desenvolvimento regional.Palavras-chave | Desenvolvimento regional; diversidade regional; potencialidades; região.Código JEL | O18; R11; R58. REGIONAL DEVELOPMENT: REGIONAL DIVERSITY AS POTENTIALAbstractThroughout the second half of the twentieth century the debate on regional development in Brazil received special attention, especially among economists and geographers. Two main approaches stand out in the debate: one that understands the promotion of regional development from the reduction / elimination of regional inequalities, and the other proposes the understanding of regional diversity and potentiality for developing regions. The first was and still remains hegemonic, being evidenced both in scientific papers and in documents guiding public policies in the area. The latter is evidenced most clearly from the 1990s on, a period when the concept of region is resumed under a new approach. This paper aims to clarify the meaning of each of these concepts in order to contribute to the qualification of the discussion on regional development.Keywords | Regional

  8. Central Region Regionally Ecological Significant Areas

    Data.gov (United States)

    Minnesota Department of Natural Resources — This is an analysis of regionally significant Terrestrial and Wetland Ecological Areas in the seven county metropolitan area. Individual forest, grassland and...

  9. PREVALÊNCIA DE LEUCOSE ENZOÓTICA BOVINA, DIARRÉIA VIRAL BOVINA RINOTRAQUEÍTE INFECCIOSA BOVINA E NEOSPOROSE BOVINA EM 26 PROPRIEDADES LEITEIRAS DA REGIÃO NORDESTE DO RIO GRANDE DO SUL, BRASIL PREVALENCE OF ENZOOTIC BOVINE LEUKOSIS, BOVINE VIRAL DIARRHEA, INFECTIOUS BOVINE RHINOTRACHEITIS AND BOVINE NEOSPOROSIS IN 26 DAIRY CATTLE FARMS FROM THE NORTHEAST REGION OF RIO GRANDE DO SUL , BRAZIL

    Directory of Open Access Journals (Sweden)

    Cleber Fiori

    2008-12-01

    .936 and sex (p = 0.562. Of the 360 sheep samples, nine (2.5% were reactive. There also was no significant association between the analyzed variables and the seropositiveness for brucellosis: age group (p = 0.522; race (p = 0.576 and sex (p = 0.461. Significant association was observed (p = 0.042 among the studied species and seropositiveness for brucellosis in the investigated animals. The seropositiveness for Brucella abortus in goats and sheep was traced for the first time in the “Sertão” (dry interior region, backlands of Pernambuco, fact that can hinder the success of the National Program of Control and Erradication of Brucellosis, due to the fact that it is common to raise small ruminants with bovines in this area, besides representing risks to Public Health.
     
    KEY WORDS: Brucellosis, ovines, caprines, small ruminants, serodiagnosis.

  10. EVALUATING FUNCTIONAL REGIONS

    Directory of Open Access Journals (Sweden)

    Samo Drobne

    2012-12-01

    Full Text Available In the paper, we suggest an approach to evaluate the number and composition of functional regions. Suggested approach is based on basic characteristics of functional regions, that are (1 more intensive intra-regional than the inter-regional interactions and (2 internal social and economic heterogeneity. Those characteristics are measured by factors estimated in spatial interaction model. The approach to evaluate functional regions was applied to Slovenia for three time periods.

  11. Polymorphisms at the 3' untranslated region of SLC11A1 gene are associated with protection to Brucella infection in goats.

    Science.gov (United States)

    Iacoboni, Paola A; Hasenauer, Flavia C; Caffaro, M Eugenia; Gaido, Analia; Rossetto, Cristina; Neumann, Roberto D; Salatin, Antonio; Bertoni, Emiliano; Poli, Mario A; Rossetti, Carlos A

    2014-08-15

    Goats are susceptible to brucellosis and the detection of Brucella-infected animals is carried out by serological tests. In other ruminant species, polymorphisms in microsatellites (Ms) of 3' untranslated region (3'UTR) of the solute carrier family 11 member A1 (SLC11A1) gene were associated with resistance to Brucella abortus infection. Goats present two polymorphic Ms at the 3'UTR end of SLC11A1 gene, called regions A and B. Here, we evaluated if polymorphisms in regions A and/or B are associated with Brucella infection in goats. Serum (for the detection of Brucella-specific antibodies) and hair samples (for DNA isolation and structure analysis of the SLC11A1 gene) were randomly collected from 229 adult native goats from the northwest of Argentina. Serological status was evaluated by buffer plate antigen test (BPAT) complemented by the fluorescent polarization assay (FPA), and the genotype of the 3'UTR of the SLC11A1 gene was determined by capillary electrophoresis and confirmed by sequence analysis. Polymorphisms in regions A and B of the 3'UTR SLC11A1 gene were found statistically significant associated with protection to Brucella infection. Specifically, the association study indicates statistical significance of the allele A15 and B7/B7 genotype with absence of Brucella-specific antibodies (p=0.0003 and 0.0088, respectively). These data open a promising opportunity for limiting goat brucellosis through selective breeding of animals based on genetic markers associated with natural resistance to B. melitensis infection.

  12. CONCEPTS OF REGIONALIZATION, REGIONAL SUBSYSTEMS, REGIONAL COMPLEXES AND REGIONAL TRANSFORMATIONS IN CONTEMPORARY IR

    Directory of Open Access Journals (Sweden)

    Alexei D. Voskressenski

    2012-01-01

    Full Text Available Abstract: The article examines theoretical aspects of regional dimension in international relations and the ways to apply in practice the results of world politics’ analysis from the viewpoint of regional and macro-regional levels. It also considers the key discussion points on regions and macro-regions in the world and Russian research literature. The author concludes that this is exactly the new IR concepts of regional level which should become the prototype of original Russian regional and country studies. These studies need to have a higher degree of theoretical conceptualization of regional level as their basis. Such an approach will enable the researchers to draw deep and unconventional conclusions of practical use especially as far as the non-Western parts of the world which are traditionally less examined by Western researchers, theorists and practitioners are concerned.

  13. Drycleaner Database - Region 7

    Data.gov (United States)

    U.S. Environmental Protection Agency — THIS DATA ASSET NO LONGER ACTIVE: This is metadata documentation for the Region 7 Drycleaner Database (R7DryClnDB) which tracks all Region7 drycleaners who notify...

  14. Redefining Regional Economic Layout

    Institute of Scientific and Technical Information of China (English)

    Yan Manman

    2010-01-01

    @@ In 2009, the Chinese government approved planning for 9 regional development zones, including plans for the Taiwan Strait West Bank Economic Zone, the Guanzhong-Tianshui Economic Zone, the Jiangsu Coastal Region, the Tumen River Region and Yellow River High-efficient Ecological Economic Zone, among others.

  15. Redefining Regional Economic Layout

    Institute of Scientific and Technical Information of China (English)

    Yan Manman

    2011-01-01

    @@ In 2009, the Chinese government approved planning for 9 regional development zones, including plans for the Taiwan Strait West Bank Economic Zone, the Guanzhong-Tianshui Economic Zone, the Jiangsu Coastal Region, the Tumen River Region and Yellow River High-efficient Ecological Economic Zone, among others.

  16. Balancing Regional Development

    Institute of Scientific and Technical Information of China (English)

    LAN XINZHEN

    2010-01-01

    @@ An early version of a regional economic plan for the Xinjiang Uygur Autonomous Region is under scrutiny by the State Council,China's cabinet.The draft,which could be passed and implemented by May,will focus on the energy,tourism,iron and steel,new agriculture and recycling economy sectors in the autonomous region.

  17. The Regional Dimension

    DEFF Research Database (Denmark)

    Eskjær, Mikkel Fugl

    2013-01-01

    is largely dependent on regional media systems, yet the role this regional dimension plays has been largely overlooked. This article presents a comparative study of climate-change coverage in three geo-cultural regions, The Middle East, Scandinavia, and North America, and explores the link between global...

  18. Screening and detection of repetitive extragenic palindromic sequences with TLR9 agonistic activity from Brucella abortus DNA%活化TLR9的牛种布氏菌基因外重复回文序列筛选及活性检测

    Institute of Scientific and Technical Information of China (English)

    张雅娴; 白丽云; 王占黎; 王英; 于慧

    2016-01-01

    目的 筛选具有活化Toll样受体9(TLR9)的牛种布氏菌基因外重复回文序列(Repetitive Extragenic Palin-drome Squence,REPs)并检测其活性,为布氏菌病的治疗提供新思路.方法 基于Brucella abortus A13334基因组序列,利用生物信息学技术识别其REPs后,合成序列.将合成的天然骨架的脱氧寡核苷酸(ODNs)转染小鼠单核巨噬细胞株RAW264.7,以ELISA检测IFN-α的分泌水平.采用TLR9-siRNA沉默RAW264.7中的TLR9,将上述诱导IFN-α分泌增加的阳性ODN序列转染RAW264.7,ELISA方法检测IFN-α的分泌变化.结果 筛选出1857条牛种布氏菌REPs,选择2级茎环结构较好的5条ODNs序列进行合成,ELISA方法检测显示ODNs M4、M5介导IFN-α分泌量显著高于阴性对照(P<0.05),且阳性ODN M5所介导的IFN-α分泌可以被TLR9-siRNA显著抑制.结论 布氏菌基因组中存在可以活化TLR9信号通路的REPs,此结果有助于对布氏菌致病和免疫机制的认识.

  19. Local, Regional or Global?

    DEFF Research Database (Denmark)

    Geisler Asmussen, Christian

    This paper proposes a multidimensional index of regional and global orientation which can be used in confirmatory studies with econometric methodologies. Unlike extant measures, the index is objectively scaled and controls for home country orientation and market size differences. The index is shown...... in fact reflects strong home country biases.Keywords: globalization; regional integration; global strategy; regional strategy; local strategy; triad; liability of foreignness...... to be consistent with models of internationalization that incorporate different assumptions about strategic choice and global competition. Preliminary results show that large multinationals follow home region oriented internationalization paths, although much of the regional effect reported by previous studies...

  20. Evolving Regional Security in the Andean Region

    Science.gov (United States)

    2004-03-19

    security forces or judicial systems reduces the effectiveness of regional cooperation. Past efforts to achieve consensus among the Andean states have...Trimestre 2001. Nuñez, Joseph. Una Arquitectura para la Seguridad del Siglo XXI par alas Américas: Cooperación Multilateral, Paz y Poder Flexible

  1. Connecting to Regional Markets?

    DEFF Research Database (Denmark)

    Coulibaly, Souleymane; Thomsen, Lotte

    2016-01-01

    Central Asian food processors face a number of constraints when they attempt to export to the region and beyond. The Central Asian economies in focus here are landlocked, and thus lack easy access to sea transport. In addition, the region's transport network was built to reinforce the interdepend......, and makes a novel attempt to examine how these factors lead to challenges for local food processing producers trying to sell their products in the region and beyond....

  2. Entrepreneurship and regional development

    DEFF Research Database (Denmark)

    Müller, Sabine

    This literature review examines how entrepreneurship and regional development has been previously addressed theoretically and empirically. Regional Science and Entrepreneurship are two fields with their own distinct literature's. The question is therefore, how do these two fields talk about...... the respective other? What are the commonalities and differences? The purpose of this article is to create an analytical synthesis by combining the insights of the two literature's in order to gain a fuller understanding of the relation between entrepreneurship and regional development....

  3. Border region studies

    DEFF Research Database (Denmark)

    Makkonen, Teemu; Williams, Allan

    2016-01-01

    The contemporary conditions of academic capitalism exert pressures on researchers to avoid ‘peripheral’ journals and ‘unfashionable’ topics. Here an attempt is made to shed light onto the structure of one such ‘offbeat’ field, namely ‘border region studies’, by discussing its geographical...... distribution, key themes, significance and impact. The review suggests that border region studies can be considered a significant and important ‘branch’ of regional studies, which accounts for a small but increasing proportion of regional studies research particularly in Europe and North America. Four main...

  4. Regional Redistribution and Migration

    DEFF Research Database (Denmark)

    Manasse, Paolo; Schultz, Christian

    We study a model with free migration between a rich and a poor region. Since there is congestion, the rich region has an incentive to give the poor region a transfer in order to reduce immigration. Faced with free migration, the rich region voluntarily chooses a transfer, which turns out...... to be equal to that a social planner would choose. Provided migration occurs in equilibrium, this conclusion holds even in the presence of moderate mobility costs. However, large migration costs will lead to suboptimal transfers in the market solution...

  5. Regions in Slovenia

    Directory of Open Access Journals (Sweden)

    Andrej Čokert

    1999-12-01

    Full Text Available The reasons behind the need to prepare a law on regions are both internal and external in nature. We need regions as a second level of local government primarily to counter internal development problems and the need for decentralisation in Slovenia. Developmentaly stagnant and depressed areas account for more than 70% of Slovene territory. The share of founds earmarked directly from central government for regional promotion is falling and is lower than the average in European Union countries. Analyses of population and employment, and of the economic, infrastructure and educational capacities of the Slovene regions reveal serious regional differences. The reasons for the establishment of regions in Slovenia also lie in the diversity of regional problems, which are different in Zasavje, Pomurje, Gorenjska or Primorska. Any restriction to an administrative territorial division would blur the special regional features which, even by Europe-wide comparision, are characteristic of Slovenia. And we cannot simplify the tackling of urgent regional problems as being a matter for relations between the central government and a large number of very diverse municipalities.

  6. Regional Innovation Clusters

    Data.gov (United States)

    Small Business Administration — The Regional Innovation Clusters serve a diverse group of sectors and geographies. Three of the initial pilot clusters, termed Advanced Defense Technology clusters,...

  7. Regional Size, Wealth and EU Regional Policy

    Directory of Open Access Journals (Sweden)

    José M. Pavía

    2012-01-01

    Full Text Available Los fondos de convergencia tienen como objetivo estimular el crecimiento de las regiones menos desarrolladas dentro de la Unión Europea. Para que una región pueda ser elegida debe ser catalogada como NUTS-2. Sin embargo, como es bien conocido, el valor que toma cualquier variable no es independiente de la escala espacial (y temporal utilizada. El objetivo de este artículo es mostrar la gran sensibilidad que muestra el criterio utilizado en la UE para seleccionar regiones de convergencia y evaluar el impacto que tendría en la distribución del presupuesto entre los diferentes países la utilización de una división territorial diferente.

  8. toward star forming regions

    Directory of Open Access Journals (Sweden)

    Evan Jordan

    2008-01-01

    Full Text Available En este artículo se reportan observaciones de la línea J= 1 → 0 de 12CS en 20 regiones de formación estelar utilizando el telescopio Haystack de 37m del MIT. También se observaron tres regiones en la línea J= 1 → 0 de 13CS, y cinco regiones en la transición de 44 GHz de CH3OH (JK = 70 → 61 A+. Las emisiones de 13CS y CH3OH fueron detectadas en todas las regiones observadas, y la emisión de 12CS fue detectada en 19 de las 20 regiones, 11 de las cuales son nuevas detecciones. Se encontró una alta correlación entre el ancho de la línea de 12CS y la distancia a las regiones, lo que indica que la emisión de 12CS J= 1 → 0 podrá ser utilizada para estimar las distancias a regiones de formación estelar. Asimismo se reporta la detección de variabilidad de los máseres de CH3OH (44 GHz en la mayoría de las regiones observadas.

  9. Emergence of regional clusters

    DEFF Research Database (Denmark)

    Dahl, Michael S.; Østergaard, Christian Richter; Dalum, Bent

    2010-01-01

    The literature on regional clusters has increased considerably during the last decade. The emergence and growth patterns are usually explained by such factors as unique local culture, regional capabilities, tacit knowledge or the existence of location-specific externalities (knowledge spillovers,...

  10. The Scandinavian regional model

    DEFF Research Database (Denmark)

    Torfing, Jacob; Lidström, Anders; Røiseland, Asbjørn

    2015-01-01

    This article maps how the sub-national regional levels of governance in Denmark, Norway and Sweden have changed from a high degree of institutional convergence to a pattern of institutional divergence. It analyses the similarities and differences in the changes in regional governance and discusses...

  11. Identification of a single-nucleotide insertion in the promoter region affecting the sodC promoter activity in Brucella neotomae.

    Directory of Open Access Journals (Sweden)

    Dina A Moustafa

    Full Text Available Brucella neotomae is not known to be associated with clinical disease in any host species. Previous research suggested that B. neotomae might not express detectable levels of Cu/Zn superoxide dismutase (SOD, a periplasmic enzyme known to be involved in protecting Brucella from oxidative bactericidal effects of host phagocytes. This study was undertaken to investigate the genetic basis for the disparity in SOD expression in B. neotomae. Our Western blot and SOD enzyme assay analyses indicated that B. neotomae does express SOD, but at a substantially reduced level. Nucleotide sequence analysis of region upstream to the sodC gene identified a single-nucleotide insertion in the potential promoter region. The same single-nucleotide insertion was also detected in the sodC promoter of B. suis strain Thomsen, belonging to biovar 2 in which SOD expression was undetectable previously. Examination of the sodC promoter activities using translational fusion constructs with E. coli β-galactosidase demonstrated that the B. neotomae and B. suis biovar 2 promoters were very weak in driving gene expression. Site-directed mutation studies indicated that the insertion of A in the B. neotomae sodC promoter reduced the promoter activity. Increasing the level of SOD expression in B. neotomae through complementation with B. abortus sodC gene did not alter the bacterial survival in J774A.1 macrophage-like cells and in tissues of BALB/c and C57BL/6 mice. These results for the first time demonstrate the occurrence of a single-nucleotide polymorphism affecting promoter function and gene expression in Brucella.

  12. Bridging regional innovation

    DEFF Research Database (Denmark)

    Hansen, Teis

    2013-01-01

    The topics of regional innovation systems (RIS) and cross-border regions attract increasing attention, but few studies combine the themes. Further, the existing empirical studies of cross-border innovation and knowledge creation analyse one case at one point in time, thus, making it difficult...... to assess the progress of integration in the regions, as well as the effect of cross-border innovation policies. Consequently, important questions are left unanswered, including the central research question of this paper: does the sudden removal of significant physical barriers directly impacts...... collaboration activity in cross-border innovation systems? This paper examines regional integration in the Oresund Region over time. It deals with a specific part of the RIS, as it analyses research collaboration between actors from the Danish and Swedish sides, with a specific emphasis on the biotech industry...

  13. Regional Ocean Data Assimilation

    KAUST Repository

    Edwards, Christopher A.

    2015-01-03

    This article reviews the past 15 years of developments in regional ocean data assimilation. A variety of scientific, management, and safety-related objectives motivate marine scientists to characterize many ocean environments, including coastal regions. As in weather prediction, the accurate representation of physical, chemical, and/or biological properties in the ocean is challenging. Models and observations alone provide imperfect representations of the ocean state, but together they can offer improved estimates. Variational and sequential methods are among the most widely used in regional ocean systems, and there have been exciting recent advances in ensemble and four-dimensional variational approaches. These techniques are increasingly being tested and adapted for biogeochemical applications.

  14. Building Regional Competencies

    DEFF Research Database (Denmark)

    Norus, Jesper

    2004-01-01

    a dominating position in the global marketfor industrial enzymes from 1870-2004. The case of industrial enzymes shows how aregion has been able to build sustainable competitive advantages from its distinctivecompetencies. This is done through a mixture of outsourcing and in sourcing ofcompetencies, knowledge...... and technologies from other regions in a ramified set ofinteracting networks. The key personnel within the regions firms are deliberatelyallowed to engage in the formations of these non-disclosure network activities so thatprofessional knowledge communities has been established across regional boundariesand...

  15. Regional ocean data assimilation.

    Science.gov (United States)

    Edwards, Christopher A; Moore, Andrew M; Hoteit, Ibrahim; Cornuelle, Bruce D

    2015-01-01

    This article reviews the past 15 years of developments in regional ocean data assimilation. A variety of scientific, management, and safety-related objectives motivate marine scientists to characterize many ocean environments, including coastal regions. As in weather prediction, the accurate representation of physical, chemical, and/or biological properties in the ocean is challenging. Models and observations alone provide imperfect representations of the ocean state, but together they can offer improved estimates. Variational and sequential methods are among the most widely used in regional ocean systems, and there have been exciting recent advances in ensemble and four-dimensional variational approaches. These techniques are increasingly being tested and adapted for biogeochemical applications.

  16. Constructing Regional advantage

    DEFF Research Database (Denmark)

    Asheim, Bjørn T.; Boschma, Ron; Cooke, Phil

    2011-01-01

    This paper presents a regional innovation policy model based on the idea of constructing regional advantage. This policy model brings together concepts like related variety, knowledge bases and policy platforms. Related variety attaches importance to knowledge spillovers across complementary...... economic development within and between regions in action lines appropriate to incorporate the basic principles behind related variety and differentiated knowledge bases....... sectors. The paper categorizes knowledge into ‘analytical’ (science based), ‘synthetic’ (engineering based) and ‘symbolic’ (arts based) in nature, with different requirements of ‘virtual’ and real proximity mixes. The implications of this are traced for evolving ‘platform policies’ that facilitate...

  17. Complex Regional Pain Syndrome

    Science.gov (United States)

    Complex regional pain syndrome (CRPS) is a chronic pain condition. It causes intense pain, usually in the arms, hands, legs, or feet. ... in skin temperature, color, or texture Intense burning pain Extreme skin sensitivity Swelling and stiffness in affected ...

  18. Vestnorden. A functional region?

    Directory of Open Access Journals (Sweden)

    Grétar Þór Eyþórsson

    2013-06-01

    Full Text Available This article discusses the issue of what kind of a region Vestnorden is. The need for such a discussion arises from the challenges posed by globalisation for the idea and construction of the West Nordic space, and the need to observe how this regional unit counters these processes. The article is based on an analytical framework which presupposes that a functional region has to consist of four elements. First, whether the space has its own institutions for decision making; second, how far there is economic complementarities among the involved nations and territories; third, to what degree they have common economic interests; and finally, to what extent social, cultural and historical ties exist between the territories involved. The article argues that there is an urgent need to discuss and reorient the institutional functionality of Vestnorden as a regional unit.

  19. Region 9 Tribal Lands

    Data.gov (United States)

    U.S. Environmental Protection Agency — Dataset of all Indian Reservations in US EPA Region 9 (California, Arizona and Nevada) with some reservation border areas of adjacent states included (adjacent areas...

  20. Regional Air Quality Data

    Data.gov (United States)

    U.S. Environmental Protection Agency — This asset provides data on regional air quality, including trace level SO2, nitric acid, ozone, carbon monoxide, and NOy; and particulate sulfate, nitrate, and...

  1. Regional National Cooperative Observer

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA publication dedicated to issues, news and recognition of observers in the National Weather Service Cooperative Observer program. Issues published regionally...

  2. Crisis and Regional Integration

    DEFF Research Database (Denmark)

    Dosenrode, Søren

    ‘Crisis’ has been a word frequently heard of over the last couple of years, both in a global meaning (e.g. the environmental crisis, the financial crisis) and also in a more regional or national meaning, many times related to Africa (Horn of Africa, Ivory Coast, DR Congo, Zimbabwe, Ivory Coast......, Tunisia, Egypt …. ), where the crisis referred to could be humanitarian, environmental, economic, political … Europe, too, has also according to mass media, been a victim of a crisis, the financial one. Could ‘crisis’ be a beginning of enhanced regional integration? This paper will try to look...... at the processes of regional integration in relation to ‘crisis’ in Africa and Europe. First, this paper will look at the concept of ‘crisis’, before it moves on to discuss ‘regional integration’ and the correlation between the two, emphasizing the approaches of neo-functionalism and federal theory...

  3. Regional Snowfall Index (RSI)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — NOAA's National Climatic Data Center is now producing the Regional Snowfall Index (RSI) for significant snowstorms that impact the eastern two thirds of the U.S. The...

  4. Spearheading Regional Integration

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Shortly after the ASEAN Regional Forum in Kuala Lumpur, Thai Ambassador and rotating Chairman of the ASEAN Beijing Committee, Jullapong Nonsrichai, answered questions from Beijing Review reporter Yan Wei on ASEAN's role and the China-ASEAN relationship.

  5. Aeromagnetic Regional Grid Data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Several regions are represented in this unique collection of earth surface measurements of magnetic field parameters and their related anomalies. The DNAG Magnetics...

  6. Regional Entrepreneurial Scorecard

    Directory of Open Access Journals (Sweden)

    JOSÉ DANIEL LORENZO GÓMEZ

    2008-12-01

    Full Text Available This paper examines the relationship between regional development and the creation of new companies from a micro perspective of institutional approach, focusing on the most relevant stakeholders involved in the process. The contribution of entrepreneurs to regional economic growth has been subject of special attention by the authorities, what is needed for a system of references to assess the adequacy of public programs for the promotion of entrepreneurial activity. Based on the concept of Balanced Scorecard, which provides a logical structure that relates and integrates and allows indicators refl ect the interests of stakeholders in shaping the Balanced Scorecard, the authors propose a Regional Entrepreneurship Scorecard (RES, as a tool for monitoring policies and programmes aimed at promoting start-ups. This RES uses information from international GEM project, which offers a vision of reality enterprising in different countries and regions.

  7. On regional geomagnetic charts

    Science.gov (United States)

    Alldredge, L.R.

    1987-01-01

    When regional geomagnetic charts for areas roughly the size of the US were compiled by hand, some large local anomalies were displayed in the isomagnetic lines. Since the late 1960s, when the compilation of charts using computers and mathematical models was started, most of the details available in the hand drawn regional charts have been lost. One exception to this is the Canadian magnetic declination chart for 1980. This chart was constructed using a 180 degrees spherical harmonic model. -from Author

  8. Regional monitoring program

    Energy Technology Data Exchange (ETDEWEB)

    Andersen, B.V.; Soldat, J.K.

    1957-08-26

    The purpose of the Regional Monitoring program is to conduct surveys to detect, measure, and to evaluate environmental radiation, particularly that of HAPO origin. Estimations of total environmental dose and HAPO's contribution to this dose, in units of fraction of public exposure limits, are calculated. Corollary functions include the use of Regional Monitoring data to establish and predict trends in environmental exposure components, and to facilitate correlation of environmental radioactivity with plant processes, process changes, and waste disposal practices.

  9. North American Regional Report

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2007-11-15

    North America is an energy community fortunate to be endowed with a rich and varied resource base. It consumes about a third of the world's energy and produces about one quarter of world energy supply. North America depends on a mix of complementary energy sources that should remain competitive but not in conflict. The current supply mix varies between Canada, the United States and Mexico, but fossil fuels are dominant across the region, leaving the three member countries vulnerable to a myriad of risks associated with traditional supply sources. Energy trade between all three countries is also a major contributor to the region's economy. Thus, the impetus for collaboration across the region has grown out of the common goals of energy security and economic prosperity. The goal of the WEC regional group was to discuss avenues for advancing North American cooperation and coordination on a range of energy issues. An additional objective was to develop policy recommendations that will facilitate effective development and use of the region's energy resources. Results and recommendtaions are summarized from three forums that focused on the pertinent issues of energy trade, energy efficiency and energy diversification. The inaugural forum (Energy Trade) was held in Washington, D.C. in the fall of 2005. The following summer, the second forum (Energy Efficiency) took place in Mexico City. The third forum (Energy Diversification) was hosted in Halifax, Nova Scotia.

  10. Northeast Regional Biomass Program

    Energy Technology Data Exchange (ETDEWEB)

    Lusk, P.D.

    1992-12-01

    The Northeast Regional Biomass Program has been in operation for a period of nine years. During this time, state managed programs and technical programs have been conducted covering a wide range of activities primarily aim at the use and applications of wood as a fuel. These activities include: assessments of available biomass resources; surveys to determine what industries, businesses, institutions, and utility companies use wood and wood waste for fuel; and workshops, seminars, and demonstrations to provide technical assistance. In the Northeast, an estimated 6.2 million tons of wood are used in the commercial and industrial sector, where 12.5 million cords are used for residential heating annually. Of this useage, 1504.7 mw of power has been generated from biomass. The use of wood energy products has had substantial employment and income benefits in the region. Although wood and woodwaste have received primary emphasis in the regional program, the use of municipal solid waste has received increased emphasis as an energy source. The energy contribution of biomass will increase as potentia users become more familiar with existing feedstocks, technologies, and applications. The Northeast Regional Biomass Program is designed to support region-specific to overcome near-term barriers to biomass energy use.

  11. Entrepreneurship and Regional Development:

    DEFF Research Database (Denmark)

    Müller, Sabine

    of entrepreneurship). This study sets out to obtain an in-depth understanding of the micro-, community-, and regional-level localized entrepreneurial processes as well as the way in which these processes are intertwined with the spatial context. The contribution of this dissertation lies in the illustration of how...... influenced by such interactions? In approaching these questions, this dissertation focuses on why entrepreneurs act (the causes of entrepreneurship, anchored in the context), how they act (the entrepreneurial practices, action, and activities), and what happens when they act (the outcomes and impact......, culture, history, and natural resources. The insights of this thesis are believed to be vital for understanding why certain types of local entrepreneurship prevail in certain regions. This can further our knowledge of how to foster and enable entrepreneurship in lagging regions. In addition, this study...

  12. Regione “Test”

    Directory of Open Access Journals (Sweden)

    Roberto Gasparini

    2012-11-01

    Full Text Available In “Test” region 75% of women (aged 24-64 are screened regularly, meaning every 3 years. A cost-effectiveness analysis shows that, considering regional tariffs, the multiple cohort (12-year-old + 25-year-old women vaccination strategy with a 50% coverage, even if the coverage is much lower than 90%, could prevent 8 cases of cervicocarcinoma and 4 related deaths more than the vaccination of only 12-year-old girls, and thus proves to be cost-effective (8,721 €/QALY.

  13. Cold regions isotope applications

    Energy Technology Data Exchange (ETDEWEB)

    Perrigo, L.D.; Divine, T.E.

    1976-04-01

    Pacific Northwest Laboratories (PNL) started the Cold Regions Isotope Applications Program in FY-1975 to identify special conditions in the Arctic and similar geographic areas (Cold Regions) where radioisotope power, heater, or sterilization systems would be desirable and economically viable. Significant progress was made in the first year of this program and all objectives for this initial 12-month period were achieved. The major conclusions and recommendations resulting for this effort are described below. The areas of interest covered include: radiosterilization of sewage; heating of septic tanks; and radioisotope thermoelectric generators as power sources for meteorological instruments and navigational aids. (TFD)

  14. Modern regional innovation policy

    NARCIS (Netherlands)

    McCann, Philip; Ortega-Argiles, Raquel

    2013-01-01

    This paper analyses the evolution of regional innovation policy into the mainstream of public policy. The paper examines the empirical and theoretical developments which have shifted much of the focus on innovation-related issues to matters of economic geography. As well as academic material we also

  15. Promoting regional mobility

    DEFF Research Database (Denmark)

    Jensen, Anne

    Pricing of transport has been part of EU's common transport policy since this gained momentum in the early 1990s. Since then, it has been closely connected to the trans-European transport network (TEN-T) and to rising demands of efficient mobility systems at a local, regional and Community scale....

  16. Australia's Regional Youth Exodus.

    Science.gov (United States)

    Gabriel, M.

    2002-01-01

    Examines media coverage of youth outmigration from Tasmania in the context of Australia's regional crisis. Focuses on how young people are constructed by others and positioned in others' visions of their rural home towns. Discusses two recurring narratives: strategies to keep youth at home, and preoccupation with the "best and brightest"…

  17. Regionalizing global climate models

    NARCIS (Netherlands)

    Pitman, A.J.; Arneth, A.; Ganzeveld, L.N.

    2012-01-01

    Global climate models simulate the Earth's climate impressively at scales of continents and greater. At these scales, large-scale dynamics and physics largely define the climate. At spatial scales relevant to policy makers, and to impacts and adaptation, many other processes may affect regional and

  18. Benchmarks: WICHE Region 2012

    Science.gov (United States)

    Western Interstate Commission for Higher Education, 2013

    2013-01-01

    Benchmarks: WICHE Region 2012 presents information on the West's progress in improving access to, success in, and financing of higher education. The information is updated annually to monitor change over time and encourage its use as a tool for informed discussion in policy and education communities. To establish a general context for the…

  19. Platforms to Regional Economy

    Institute of Scientific and Technical Information of China (English)

    Guo Yan

    2008-01-01

    @@ Besides Canton Fair,China has many other important export and import fairs.Among them,three regional fairs,namely East China Fair(ECF),China Kunming Import & Export Commodities Fair(known as Kunming Fair),and Dalian Import & Export Commodities Fair(DIECF)ale worth the focus.

  20. Global, Local, or Regional?

    DEFF Research Database (Denmark)

    Verbeke, Alain; Geisler Asmussen, Christian

    2016-01-01

    of analysis, in addition to the country-level and the global level. Regional strategy analysis requires a fundamental rethink of mainstream theories in the international strategy sphere. This rethink involves, inter alia, internalization theory, with its resource-based view and transaction cost economics...

  1. Regionalism and geopolitics

    Directory of Open Access Journals (Sweden)

    Knežević Miloš

    2002-01-01

    Full Text Available Recognition of regional features, outlining of the contours of regions, tendency to regionalize ethnic, economic, cultural and state-administrative space, and strengthening the ideology of regionalism in the Federal Republic of Yugoslavia, that is Serbia and Montenegro, appear as a practical and political but also as a theoretical problem which includes and combines several scientific disciplines. The phenomenon of regionalism is not contradictory although it is primarily expressed through the numerous conflicts of interests rivalry and antagonisms of political subjects. The problematic side of the phenomenon of regionalism includes the result of an extremely negative and existentially tragic experience of the several years-long disintegration of the complex Yugoslav state. During the partition and disintegration of the second Yugoslavia, there also happened the disintegration of the Serbian ethnic area Growth, support and instigation of regional tendencies occurred in the historical circumstances of secession and did not stop in the post-secession period. Particularization and segmentation of political area, as well as the disintegration of the former state, did not occur in accordance with the norms of internal and international law. Legality was late and was achieved within the transformation of power reflected in the changed territorial policy of the dominant alliance of great powers. The entire past decade was characterized by an extraordinary metamorphosis of political space. Secession trend had the territorial features which included the change of borders and had been long in the focus of the global geopolitical attention. Territories were divided and made smaller. Intensive territorial dynamics within the external silhouette of the de-stated SFR of Yugoslavia resulted in the creation of several state and quasi-state political formations. Former republics became semi-sovereign states. Dispersed and displaced Serbian ethnos was configured

  2. Regional utvikling og partnerskap

    DEFF Research Database (Denmark)

    Halkier, H.; Gjertsen, A.

    2004-01-01

    of Danish regions to pursue their own agendas with regard to economic development, and explore the organizational strategies pursued by varies tiers of government in this process of rapid and profound policy change. The text is divided into three parts. The following section provides a brief outline...... of the analytical framework, based primarily on contributions from traditions within policy analysis, network theory and the new institutionalism. The main body of the text examines the changing face of spatial economic policy in Denmark, dealing in turn with the development of policies and initiatives emanating......Since the beginning of the 1990s, Danish regional policy has changed dramatically. As of January 1991, all central government incentive schemes were terminated, and since then the main components of spatial economic policy have been a host of subnational initiatives and the European Structural...

  3. Regional Stability & Peacebuilding

    DEFF Research Database (Denmark)

    It seems that regional decision makers during the last two decades has been unable to produce a sustainable peacebuilding plan for the region and it is questionable whether any remarkable change will occur in the near future. Some would argue that the political differences are simply too far apart...... continue to face, internal challenges even if agreements with a conflicting state are settled. This only underlines the necessity of initiating sustainable initiatives that are capable of affecting politicians from within, or even to some extent have the capability to bypass the political level....... With contributions from leading international scholars within the field of security studies this book sets out to explain the main security knots preventing stability to emerge and on that basis to test whether a different approach in addressing these knots. By pursuing an innovative and different approach...

  4. Banks, regions and development

    Directory of Open Access Journals (Sweden)

    Pietro Alessandrini

    2003-03-01

    Full Text Available From the 1980s onwards the banking sectors in all the industrialised countries have been experiencing intense restructuring, aggregation and consolidation, radically changing their ownership structures and geography. Whatever the reasons behind such restructuring processes, the globalisation of the credit markets, the consolidation of banking structures, the removal of barriers to the free location of banks and their penetration of peripheral markets pose two main questions. Will integration of the banking systems lead to a narrowing or a widening of the development gap between regions? What relations will there be between financial centres and the periphery, and how will financial labour be divided between national (international banks and local (regional banks? The aim of this paper is to address such questions in the light of recent developments in the theoretical and empirical literature on financial integration.

  5. Northwest Regional Climate Assessment

    Science.gov (United States)

    Lipschultz, Fred

    2011-01-01

    Objectives are to establish a continuing, inclusive National process that: 1) synthesizes relevant science and information 2) increases understanding of what is known & not known 3) identifies information needs related to preparing for climate variability and change, and reducing climate impacts and vulnerability 4) evaluates progress of adaptation & mitigation activities 5) informs science priorities 6) builds assessment capacity in regions and sectors 7) builds understanding & skilled use of findings

  6. Regional Seismic Wave Propagation

    Science.gov (United States)

    1979-07-31

    Baikal to the Pamirs, earthquakes occuring in the Baikal region, Sinkiang , the Gobi desert, southwest China and the Himalayas generated Lg/P ratios...data were obtained from stations within the USSR from earthquake events occuring in Baikal, Sinkiang , the Gobi desert, Southwest China and the...earthquakes originating in the Sinkiang province and recorded by seismo- graphic stations along the Pamir-Lena River profile [25] 0 - recorded by short

  7. Regional Renewable Energy Cooperatives

    Science.gov (United States)

    Hazendonk, P.; Brown, M. B.; Byrne, J. M.; Harrison, T.; Mueller, R.; Peacock, K.; Usher, J.; Yalamova, R.; Kroebel, R.; Larsen, J.; McNaughton, R.

    2014-12-01

    We are building a multidisciplinary research program linking researchers in agriculture, business, earth science, engineering, humanities and social science. Our goal is to match renewable energy supply and reformed energy demands. The program will be focused on (i) understanding and modifying energy demand, (ii) design and implementation of diverse renewable energy networks. Geomatics technology will be used to map existing energy and waste flows on a neighbourhood, municipal, and regional level. Optimal sites and combinations of sites for solar and wind electrical generation (ridges, rooftops, valley walls) will be identified. Geomatics based site and grid analyses will identify best locations for energy production based on efficient production and connectivity to regional grids and transportation. Design of networks for utilization of waste streams of heat, water, animal and human waste for energy production will be investigated. Agriculture, cities and industry produce many waste streams that are not well utilized. Therefore, establishing a renewable energy resource mapping and planning program for electrical generation, waste heat and energy recovery, biomass collection, and biochar, biodiesel and syngas production is critical to regional energy optimization. Electrical storage and demand management are two priorities that will be investigated. Regional scale cooperatives may use electric vehicle batteries and innovations such as pump storage and concentrated solar molten salt heat storage for steam turbine electrical generation. Energy demand management is poorly explored in Canada and elsewhere - our homes and businesses operate on an unrestricted demand. Simple monitoring and energy demand-ranking software can easily reduce peaks demands and move lower ranked uses to non-peak periods, thereby reducing the grid size needed to meet peak demands. Peak demand strains the current energy grid capacity and often requires demand balancing projects and

  8. Regional Strategic Estimate

    Science.gov (United States)

    1992-01-01

    nation’s productivity. b. Guatemala. Several leftist guerrilla groups continue to challenge Guatemala’s fragile democracy ( Organizacion Revolucionario de...light industry and textiles. Mexico’s industrial sector is much greater than that of the rest of the region. The Pan American Highway is a good...boon to the local economy, we may consume scarce resources normally available to domestic industry at its peril. Second, local resources may be rapidly

  9. Regional Shelter Analysis Methodology

    Energy Technology Data Exchange (ETDEWEB)

    Dillon, Michael B. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Dennison, Deborah [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Kane, Jave [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Walker, Hoyt [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Miller, Paul [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2015-08-01

    The fallout from a nuclear explosion has the potential to injure or kill 100,000 or more people through exposure to external gamma (fallout) radiation. Existing buildings can reduce radiation exposure by placing material between fallout particles and exposed people. Lawrence Livermore National Laboratory was tasked with developing an operationally feasible methodology that could improve fallout casualty estimates. The methodology, called a Regional Shelter Analysis, combines the fallout protection that existing buildings provide civilian populations with the distribution of people in various locations. The Regional Shelter Analysis method allows the consideration of (a) multiple building types and locations within buildings, (b) country specific estimates, (c) population posture (e.g., unwarned vs. minimally warned), and (d) the time of day (e.g., night vs. day). The protection estimates can be combined with fallout predictions (or measurements) to (a) provide a more accurate assessment of exposure and injury and (b) evaluate the effectiveness of various casualty mitigation strategies. This report describes the Regional Shelter Analysis methodology, highlights key operational aspects (including demonstrating that the methodology is compatible with current tools), illustrates how to implement the methodology, and provides suggestions for future work.

  10. Andean region study

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-10-01

    New opportunities for climate change mitigation arising from a higher energy integration among Andean Pact nations were analysed within the framework of the UNEP/GEF Project. Apart from the search for regional mitigation actions, the study was mainly aimed at detecting methodological problems which arise when passing from a strictly national view to the co-ordination of regional actions to deal with climate change. In accordance with the available resources and data, and in view of the mainly methodological nature of the project, it was decided to analyse the opportunities to delve into the energy integration of the Region as regards electricity and natural gas industries and their eventual impact on the emission of greenhouse gases. Although possibilities of setting up electricity and natural gas markets are real, their impacts on GHG emission from the energy system would not prove substantially higher than those which the nations could achieve through the use of their own energy resources, in view that the Andean systems are competitive rather than complementary. More in-depth studies and detail information will be required - unavailable for the present study - to be able to properly evaluate all benefits associated with higher energy integration. Nevertheless, the supply of natural gas to Ecuador seems to be the alternative with the highest impact on GHG emission. If we were to analyse the supply and final consumption of energy jointly, we would most certainly detect additional mitigation options resulting from higher co-operation and co-ordination in the energy field. (EHS)

  11. SERVICES AND REGIONAL DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    Sven ILLERIS

    2009-12-01

    Full Text Available The purpose of this contribution is to discuss what roles the different economic sectors, and in particular services activities (the tertiary sector play in regional development, understood as growth in production, incomes and employment in weakly developedregions. This question is approached in two ways. The contribution first contains a – primarily theoretical – re-examination of the so-called economic base model, which states that services play a passive role in regional development. The discussion leads to substantial modifications of the model. The second approach is more empirical. It will take as its point of departure the proposition – often heard, but rarely examined – that since service activities are more concentrated in big cities than other activities and in recent decades have shown higher growth rates than other economic activities, it follows that the economic development is now pulled towards big city regions. Examined by way of a statistical analysisin Denmark and France, this proposition could not be verified.

  12. OECD Reviews of Regional Innovation

    DEFF Research Database (Denmark)

    Maguire, Karen; Marsan, Giulia Ajmone; Nauwelaers, Claire;

    This book examines regional innovation in central and southern Denmark, looking at its role in the economy, its governance and policy context and regional strategies for innovation driven growth.......This book examines regional innovation in central and southern Denmark, looking at its role in the economy, its governance and policy context and regional strategies for innovation driven growth....

  13. Polymorphisms of the coding region of Slc11a1 (Nramp1 gene associated to natural resistance against bovine brucellosis

    Directory of Open Access Journals (Sweden)

    T.A. Paixão

    2012-08-01

    Full Text Available Brucelose bovina causada por Brucella abortus é uma importante doença zoonótica, caracterizada pela ocorrência de aborto durante o último trimestre da gestação, o que resulta em diminuição da fertilidade da produção de leite em vacas. A identificação de genes associados à resistência natural contra brucelose tem sido investigada com o objetivo de selecionar animais resistentes à doença. Em bovinos, é controversa a resistência natural contra B. abortus associada ao polimorfismo da região 3' UTR do gene Slc11A1 (Nramp1. Polimorfismos localizados na sequência codificadora de Slc11A1 têm sido identificados em bovinos, contudo a influência sobre a resistência natural contra brucelose não é conhecida. No presente estudo, três novos polimorfismos do gene Slc11A1 foram genotipados por análise conformacional de fita simples em vacas experimentalmente ou naturalmente infectadas por B. Abortus, e foram avaliadas a frequência de cada genótipo e sua associação com o fenótipo de resistência ou susceptibilidade à brucelose bovina. Os resultados deste estudo demonstram que alguns genótipos foram mais frequentes em animais considerados fenotipicamente susceptiveis à brucelose.

  14. Regional migratory osteoporosis

    Energy Technology Data Exchange (ETDEWEB)

    Cahir, John G. [Department of Radiology, Norfolk and Norwich University Hospital, Colney Lane, Norwich, Norfolk NR4 7UY (United Kingdom)], E-mail: john.cahir@nnuh.nhs.uk; Toms, Andoni P. [Department of Radiology, Norfolk and Norwich University Hospital, Colney Lane, Norwich, Norfolk NR4 7UY (United Kingdom)

    2008-07-15

    Regional migratory osteoporosis (RMO) is an uncommon disease characterised by a migrating arthralgia involving the weight bearing joints of the lower limb. The typical imaging findings on radiographs, magnetic resonance imaging, computed tomography and bone scintigraphy are described and illustrated. Men in their fifth and sixth decades of life are most commonly affected. The most common presentation is with proximal to distal spread in the lower limb. The world literature has been reviewed which has revealed 63 documented cases of regional osteoporosis or bone marrow oedema with migratory symptoms. Most of these cases have not been labelled as RMO and therefore the condition is probably under-diagnosed. The radiology of RMO is indistinguishable from transient osteoporosis of the hip (TOH) except for the migratory symptoms and the two conditions are likely to be part of the same spectrum of disease. Systemic osteoporosis is a more recently recognised accompanying feature that hints at an underlying aetiology and an approach to the management of this condition.

  15. Regiones cognitivas en Colombia

    Directory of Open Access Journals (Sweden)

    Robayo Camilo Alberto

    1996-06-01

    Full Text Available

    Como parte del proyecto en curso: "regiones cognitivas en Colombia", respaldado por Colciencias y el Centro de Investigaciones de la Universidad Pedagógica, este libro constituye el primer informe y sienta las bases para la segunda etapa en la que se están examinando las correlaciones entre estilos cognitivos, comprensión de lenguaje y procesamiento matemático. Redactado en estilo transparente y conciso, este libro asume el reto de explorar y dar consistencia experimental a una regionalización ecocultural de Colombia, enfocándose desde la problemática de la educación. El bajo nivel en logros específicos por área curricular y la concentración de dicha situación por regiones en el país permiten que esta investigación se proyecte desde el inicio como una crítica a los modelos pedagógicos vigentes y una propuesta para su renovación.

  16. Regional Healthcare Effectiveness

    Directory of Open Access Journals (Sweden)

    Olga Vladimirovna Kudelina

    2016-03-01

    Full Text Available An evaluation of healthcare systems effectiveness of the regions of the Russian Federation (federal districts was conducted using the Minmax method based on the data available at the United Interdepartmental Statistical Information System. Four groups of components (i.e. availability of resources; use of resources; access to resources and medical effectiveness decomposed into 17 items were analyzed. The resource availability was measured by four indicators, including the provision of doctors, nurses, hospital beds; agencies providing health care to the population. Use of resources was measured by seven indicators: the average hospital stay, days; the average bed occupancy, days; the number of operations per 1 physician surgical; the cost per unit volume of medical care: in outpatient clinics, day hospitals, inpatient and emergency care. Access to the resources was measured by three indicators: the satisfaction of the population by medical care; the capacity of outpatient clinics; the average number of visits to health facility. The medical effectiveness was also measured by three indicators: incidence with the "first-ever diagnosis of malignancy"; life expectancy at birth, years; the number of days of temporary disability. The study of the dynamics of the components and indexes for 2008–2012 allows to indicate a multidirectional influence on the regional healthcare system. In some federal districts (e.g. North Caucasian, the effectiveness decreases due to resource availability, in others (South, North Caucasian — due to the use of resources, in others (Far Eastern, Ural — due to access to resources. It is found that the effectiveness of the healthcare systems of the federal districts differs significantly. In addition, the built matrix proves the variability the of effectiveness (comparison of expenditures and results of healthcare systems of the federal districts of the Russian Federation: the high results can be obtained at high costs

  17. Strengthening regional safeguards

    Energy Technology Data Exchange (ETDEWEB)

    Palhares, L.; Almeida, G.; Mafra, O. [Brazilian-Argentine Agency for Accounting and Control of Nuclear Materials, Rio de Janeiro (Brazil)] [and others

    1996-08-01

    Nuclear cooperation between Argentina and Brazil has been growing since the early 1980`s and as it grew, so did cooperation with the US Department of Energy (DOE). The Brazilian-Argentine Agency for Accounting and Control of Nuclear Materials (ABACC) was formed in December 1991 to operate the Common System of Accounting and Control of Nuclear Materials (SCCC). In April 1994, ABACC and the DOE signed an Agreement of Cooperation in nuclear material safeguards. This cooperation has included training safeguards inspectors, exchanging nuclear material measurement and containment and surveillance technology, characterizing reference materials, and studying enrichment plant safeguards. The goal of the collaboration is to exchange technology, evaluate new technology in Latin American nuclear facilities, and strengthen regional safeguards. This paper describes the history of the cooperation, its recent activities, and future projects. The cooperation is strongly supported by all three governments: the Republics of Argentina and Brazil and the United States.

  18. Regional transmission subsystem planning

    Energy Technology Data Exchange (ETDEWEB)

    Costa Bortoni, Edson da [Quadrante Softwares Especializados Ltda., Itajuba, MG (Brazil); Bajay, Sergio Valdir; Barros Correia, Paulo de [Universidade Estadual de Campinas, SP (Brazil). Faculdade de Engenharia Mecanica; Santos, Afonso Henriques Moreira; Haddad, Jamil [Escola Federal de Engenharia de Itajuba, MG (Brazil)

    1994-12-31

    This work presents an approach for the planning of transmission systems by employing mixed--integer linear programming to obtain a cost and operating characteristics optimized system. The voltage loop equations are written in a modified form, so that, at the end of the analysis, the model behaves as a DC power flow, with the help of the two Kirchhoff`s laws, exempting the need of interaction with an external power flow program for analysis of the line loading. The model considers the occurrence of contingencies, so that the final result is a network robust to the most severe contingencies. This whole technique is adapted to the regional electric power transmission subsystems. (author) 9 refs., 4 figs.

  19. Moldova. Historic regional conference.

    Science.gov (United States)

    Moshin, V

    1995-05-01

    The Directorate of Maternal and Child Health and the Family Planning Association of Moldova organized a regional conference, which was held October 18-19, 1994, in Kishinev, Moldova, with the support of the United Nations Population Fund (UNFPA), the World Health Organization (WHO), and the International Planned Parenthood Federation (IPPF). The conference,"Problems of Family Planning in Eastern Europe," was attended by approximately 400 Moldovan delegates of governmental and nongovernmental organizations (NGOs), and by 25 delegates from Romania, Russia, Belarus, the Ukraine, and Georgia. The President of Moldova and the Ministry of Public Health of Moldova gave their approval. The main objectives of the conference were to inform the public about the recommendations of the ICPD, to analyze the status of women's reproductive health and family planning in Eastern Europe, and to find ways of implementing the ICPD Plan of Action. Major problems identified during the conference were: 1) the social and economic problems facing most families; 2) the high rate of morbidity and mortality; 3) the decrease in birth rate; 4) the increase in abortions; 5) the rising incidence of venereal disease; and 6) the absence of an effective family planning system. It was agreed that cooperation between governments and NGOs is essential in designing population programs for each country. The following goals were set: 1) to provide populations with sufficient contraceptives; 2) to actively promote family planning concepts through the mass media; 3) to train specialists and to open family planning offices and centers; 4) to introduce sex education in the curricula of Pedagogical Institutes; and 5) to create national and regional statistical and sociological databases on population issues.

  20. EPA Regional Boundaries (EPA.EPA_REGIONS) GIS Layer

    Data.gov (United States)

    U.S. Environmental Protection Agency — Each EPA Regional Office is responsible within its states for the execution of the Agency's programs. EPA has ten regional offices, each of which is responsible for...

  1. Case report 469: Spondylitis (lumbar spine) due to Brucella abortus

    Energy Technology Data Exchange (ETDEWEB)

    Manaster, B.J.

    1988-03-01

    The current case is interesting in that, although the plain radiographs were diagnostic of infection and the patient's work history suggested brucellosis, both the negative serum antibody titers to brucella and the CT appearance of large calcified psoas abscesses made the diagnosis of tuberculous spondylitis most probable. Open biopsy with tissue culture proved brucella. From this experience it appears that the presence of large calcified psoas abscesses should not eliminate the diagnosis of brucella spondylitis in the proper clinical setting.

  2. Meningen over abortus in West-Europa, 1981-2000

    OpenAIRE

    Need, A.; Ultee, W.C.; Levels, M.; Tienen, M. van

    2008-01-01

    Summary Abortion attitudes in Western Europe, 1981-2000 This article answers descriptive and explanatory questions about abortion attitudes in Western Europe between 1981 and 2000. It explains country differences and trends in these attitudes with abortion laws of states, with the norms that prevail in churches and schools, and with opinions and behaviour of others in a society. Using the European and World Value Surveys, complemented with abortion statistics and information about abortion la...

  3. Årsager, udredning og behandling af abortus habitualis

    DEFF Research Database (Denmark)

    Larsen, Elisabeth Clare; Christiansen, Ole Bjarne

    2012-01-01

    Recurrent miscarriage (RM) is defined as three or more consecutive early pregnancy losses and affects 1% of couples. RM is probably a multifactorial disease and evaluation includes parental karyotyping, testing for uterine anomalies, thyroid dysfunction, thrombophilia and immune factors. Couples...

  4. Genetiske risikofaktorer for abortus habitualis og obstetriske komplikationer

    DEFF Research Database (Denmark)

    Christiansen, Ole Bjarne

    2010-01-01

    Maternal risk factors dominate among younger women with multiple miscarriages. In all patients, genetic variants associated with low plasma mannose binding lectin or thrombophilia affect the prognosis negatively. In women with secondary recurrent miscarriage, the HLA-DRB1*03 allele or homozygocity...

  5. Meningen over abortus in West-Europa, 1981-2000

    NARCIS (Netherlands)

    Need, A.; Ultee, W.C.; Levels, M.; Tienen, M. van

    2008-01-01

    This article answers descriptive and explanatory questions about abortion attitudes in Western Europe between 1981 and 2000. It explains country differences and trends in these attitudes with abortion laws of states, with the norms that prevail in churches and schools, and with opinions and behaviou

  6. Meningen over abortus in West-Europa, 1981-2000

    NARCIS (Netherlands)

    Need, Ariana; Ultee, Wouter; Levels, Mark; Tienen, Marike van

    2008-01-01

    Summary Abortion attitudes in Western Europe, 1981-2000 This article answers descriptive and explanatory questions about abortion attitudes in Western Europe between 1981 and 2000. It explains country differences and trends in these attitudes with abortion laws of states, with the norms that prevail

  7. Challenges Towards Sustainable Tourism for Regional Development of Vlore Region

    OpenAIRE

    Aleks Prifti; Engjellushe Zenelaj

    2013-01-01

    Vlora has great potential for regional development. Tourism can be as a catalyst for theeconomic growth but still have many challenges for sustainable tourism regionaldevelopment. Regional development is one ofthe most important dimensions and key ofEU policy, which is designedto: ensure convergence between the different member statesand regions; ensure regional competitiveness and employment;ensure territorialcooperation. Vlora has all the potential to bean attractive tourismdestination. Res...

  8. Regional brain hypometabolism is unrelated to regional amyloid plaque burden

    Science.gov (United States)

    Altmann, Andre; Ng, Bernard; Landau, Susan M.; Jagust, William J.

    2015-01-01

    See Sorg and Grothe (doi:10.1093/brain/awv302) for a scientific commentary on this article. In its original form, the amyloid cascade hypothesis of Alzheimer’s disease holds that fibrillar deposits of amyloid are an early, driving force in pathological events leading ultimately to neuronal death. Early clinicopathological investigations highlighted a number of inconsistencies leading to an updated hypothesis in which amyloid plaques give way to amyloid oligomers as the driving force in pathogenesis. Rather than focusing on the inconsistencies, amyloid imaging studies have tended to highlight the overlap between regions that show early amyloid plaque signal on positron emission tomography and that also happen to be affected early in Alzheimer’s disease. Recent imaging studies investigating the regional dependency between metabolism and amyloid plaque deposition have arrived at conflicting results, with some showing regional associations and other not. We extracted multimodal neuroimaging data from the Alzheimer’s disease neuroimaging database for 227 healthy controls and 434 subjects with mild cognitive impairment. We analysed regional patterns of amyloid deposition, regional glucose metabolism and regional atrophy using florbetapir (18F) positron emission tomography, 18F-fluordeoxyglucose positron emission tomography and T1-weighted magnetic resonance imaging, respectively. Specifically, we derived grey matter density and standardized uptake value ratios for both positron emission tomography tracers in 404 functionally defined regions of interest. We examined the relation between regional glucose metabolism and amyloid plaques using linear models. For each region of interest, correcting for regional grey matter density, age, education and disease status, we tested the association of regional glucose metabolism with (i) cortex-wide florbetapir uptake; (ii) regional (i.e. in the same region of interest) florbetapir uptake; and (iii) regional florbetapir uptake

  9. American Red Cross Chapter Regions

    Data.gov (United States)

    Department of Homeland Security — The Regions are part of the national field level structure to support chapters. The Regions role is admistrative as well as provides oversight and program technical...

  10. Distances to star forming regions

    CERN Document Server

    Loinard, Laurent

    2014-01-01

    The determination of accurate distances to star-forming regions are discussed in the broader historical context of astronomical distance measurements. We summarize recent results for regions within 1 kpc and present perspectives for the near and more distance future.

  11. Regional Employment Growth, Shocks and Regional Industrial Resilience

    DEFF Research Database (Denmark)

    Holm, J.R.; Østergaard, Christian Richter

    2015-01-01

    The resilience of regional industries to economic shocks has gained a lot of attention in evolutionary economic geography recently. This paper uses a novel quantitative approach to investigate the regional industrial resilience of the Danish information and communication technology (ICT) sector...... to the shock following the burst of the dot.com bubble. It is shown that regions characterized by small and young ICT service companies were more adaptable and grew more than others, while diversity and urbanization increased the sensitivity to the business cycle after the shock. Different types of resilient...... regions are found: adaptively resilient, rigidly resilient, entrepreneurially resilient and non-resilient regions....

  12. Regional employment growth, shocks and regional industrial resilience

    DEFF Research Database (Denmark)

    Holm, Jacob Rubæk; Østergaard, Christian Richter

    2013-01-01

    The resilience of regional industries to economic shocks has gained a lot of attention in evolutionary economic geography recently. This paper uses a novel quantitative approach to investigate the regional industrial resilience of the Danish ICT sector to the shock following the burst of the dot......-com bubble. It is shown that regions characterised by small and young ICT service companies were more adaptable and grew more than others, while diversity and urbanisation increased the sensitivity to the business cycle after the shock. Different types of resilient regions are found: adaptively resilient......, rigidly resilient, entrepreneurially resilient and non-resilient regions....

  13. Sudurnes Regional Heating Corp.

    Energy Technology Data Exchange (ETDEWEB)

    Lienau, P.J. [ed.

    1996-11-01

    The Svartsengi geothermal area is close to the town of Grindavik on the Rekjanes peninsula and is part of an active fissure swarm, lined with crater-rows and open fissures and faults. The high-temperature area has an area of 2 sq. km and shows only limited signs of geothermal activity at the surface. The reservoir, however, contains lots of energy and at least 8 wells supply the Svartsengi Power Plant with steam. The steam is not useable for domestic heating purposes so that heat exchangers are used to heat cold groundwater with the steam. Some steam is also used for producing 16.4 MW{sub e} of electrical power. The article shows the distribution system piping hot water to nine towns and the Keflavik International Airport. The effluent brine from the Svartsengi Plant is disposed of into a surface pond, called the Blue Lagoon, popular to tourists and people suffering from psoriasis and other forms of eczema seeking therapeutic effects from the silica rich brine. This combined power plant and regional district heating system (cogeneration) is an interesting and unique design for the application of geothermal energy.

  14. regional economic development

    Directory of Open Access Journals (Sweden)

    Robert J. Stimson

    2005-01-01

    Full Text Available Este artículo propone que un círculo virtuoso para el desarrollo sostenible de una ciudad o región se logra a través de un proceso por el que el liderazgo proactivo y fuerte y las instituciones efectivas refuerzan la capacidad y aptitud de un lugar para hacer un mejor uso de sus dotaciones de recursos y lograr un mercado mejorado que consiga ser competitivo y emprendedor. Se propone que el funcionamiento de una ciudad o región en un punto en el tiempo y la trayectoria de su desarrollo económico a lo largo del tiempo puede representarse por su posición en un Cubo de resultados de Competitividad Regional. El artículo propone una nueva estructura de modelo donde el desarrollo económico y funcionamiento de una ciudad o región sea un resultado dependiente de cómo sus dotaciones de recursos y mercado ajustadas como variables cuasi-independientes son mediadas por la interacción entre el liderazgo, instituciones y espíritu empresarial como variables intervinientes. Las experiencias de un cierto número de casos de estudio de ciudades dentro de una variedad de lugares en EE.UU., Europa, Asia y Australia se exploran dentro de esa estructura.

  15. Regional odontodysplasia: case report

    Directory of Open Access Journals (Sweden)

    Ana Carolina Magalhães

    2007-12-01

    Full Text Available Regional odontodysplasia (RO is a rare developmental anomaly involving both mesodermal and ectodermal dental components in a group of contiguous teeth. It affects the primary and permanent dentitions in the maxilla and mandible or both jaws. Generally it is localized in only one arch. The etiology of this dental anomaly is uncertain. Clinically, affected teeth have an abnormal morphology, are soft on probing and typically discolored, yellow or yellowish-brown. Radiographically, the affected teeth show a "ghostlike" appearance. This paper reports the case of a 5-year-old girl presenting this rare anomaly on the left side of the maxillary arch, which crossed the midline. The primary maxillary left teeth (except for the canine and the primary maxillary right central incisor were missing due to previous extractions. The permanent teeth had a "ghostlike" appearance radiographically. The treatment performed was rehabilitation with temporary partial acrylic denture and periodic controls. In the future, the extraction of affected permanent teeth and rehabilitation with dental implants will be evaluated. The presentation of this case adds valuable information to pediatric dentists to review special clinical and radiographic features of RO, which will facilitate the diagnosis and treatment of patients with this condition.

  16. Microbiological and serological monitoring in hooded crow (Corvus corone cornix in the Region Lombardia, Italy

    Directory of Open Access Journals (Sweden)

    Guido Grilli

    2010-01-01

    Full Text Available The health status of 276 hooded crows (Corvus corone cornix from various provinces of Lombardy was monitored for three years. Bacteriological examination detected E. coli (76%, Campylobacter jejuni (17%, Salmonella typhimurium (11.6%, Yersinia spp. (6.5%, Clamydophila abortus and C. psittaci (2.6%; from six birds showing severe prostration Pasteurella multocida was isolated. Virological and serological tests were negative for Avian Influenza virus (AIV, West Nile virus (WNV and only three samples were positive for Newcastle disease virus (NDV but only at serology (titre 1:16.

  17. Thinking regionally: narrative, the medical humanities and region.

    Science.gov (United States)

    Waddington, Keir

    2015-06-01

    Drawing on multiple literatures from history, geography, anthropology, sociology and literature, this essay asks questions about what we mean by region and why narratives of region should matter to the medical humanities. The essay surveys how region can be used as a lens of analysis, exploring the various academic approaches to region and their limitations. It argues that regions are dynamic but also unstable as a category of analysis and are often used uncritically by scholars. In encouraging scholars working in the medical humanities to be aware that regions are not simple objective or analytical boxes, the essay shows how an awareness of region helps challenge metropolitan whiggism and ideas of core and periphery to give a more prominent place to hinterlands, market towns and rural environments. Furthermore, the essay considers how incorporating region into our understanding of illness can offer new insights. It demonstrates the need for scholars to be attuned to the narratives constructed around regions, suggesting that regions can be viewed as discursive formations that provide a frame for understanding both collective and personal ideas of, and responses to, health and illness, disease and healing, to create what Megan Davies calls a more nuanced 'intellectual cartography'.

  18. Caspian Region in the System of International Regions

    Directory of Open Access Journals (Sweden)

    Kosov Gennadiy Vladimirovich

    2015-12-01

    Full Text Available The given research paper presents an attempt to analyze the Caspian region from the point of view of its cross-border specifics. In contrast to the traditional understanding of the Caspian region as the region of heightened geopolitical significance the authors analyze its peculiarities and potential in the context of the cross-border cooperation dynamics. It is especially emphasized that the Caspian region may be considered as a cross-border region which, despite the substantial similarities is different from international transnational regions. The authors focus attention on a key contradiction of the Caspian region. On the one hand, national interests and foreign policy of the states-members of the Caspian region (Azerbaijan, Iran, Kazakhstan, Russia and Turkmenistan have a national country and regional importance, since their implementation will form an effective regional security system. On the other hand, the Caspian region attracts the attention of the world powers, becoming a subject of their interest, as it is of great importance for world energy markets and has status of a transit zone between Europe and Asia. For Azerbaijan, Iran, Kazakhstan, Russia, Turkmenistan the Caspian status is combined with their most important characteristic of post-Soviet states - shared long historical stage of development within the Soviet Union. The article distinguished and analyzed four main periods of establishment of cross-border cooperation in the Caspian region. The national specificity of modernization reforms in all the countries of the Caspian region have interrelations with further development of cross-border cooperation, the priority of that performs convergence of Caspian states on a number of strategic issues, including economic cooperation and security.

  19. A Declining Region: Provincial Renaissance Revisited (Case of Volgograd Region

    Directory of Open Access Journals (Sweden)

    Drozdova Yuliya

    2014-12-01

    Full Text Available The paper analyzes findings yielded by the empirical study performed in the framework of an RGNF grant entitled “Image of a region as a communicative strategy for the authorities and mass media”. The methods of study included expert survey and formal survey (N=1000, July-September 2013, studying the opinion of respondents who were either internal experts residing in the Volgograd region (N=20; May-September 2013 or external experts who reside outside the region but maintain stable ties with the representatives of state and municipal authorities, regional mass media and business. The findings indicate that the Volgograd region has fallen behind other modernized Russian regions, that young people tend to leave it, that a negative image of the region as a declining territory persists. Answers to the open question “What is unacceptable for you in the existing image of the Volgograd region?” revealed major problems determining the local context of a declining region, and those were issues associated with inefficient regional/municipal administration: “the condition of the roads”, “constant replacement of people in the administration”, “politics as a whole”, “a destitute region without a good manager”, “unemployment”, “countryside is dying off”, “indifference of the authorities”, “roads, housing and public utilities and the administration”, “the authorities are not responsible for the people”, “the authorities do not solve the problems of the city or its people”, “thieving”, “dishonest authorities”, “the region goes to rack and ruin, no kindergartens or jobs”, “one cannot even walk in the streets”, “corruption”, “a stagnant region with low pay”, “no perspectives in the future”, “the region is stagnating due to corruption among officials”. According to the local Census Bureau, the Volgograd region can be classified as a declining territory where the population decline

  20. Regional governance: strategies and disputes in health region management

    Directory of Open Access Journals (Sweden)

    Adriano Maia dos Santos

    2014-08-01

    Full Text Available OBJECTIVE To analyze the regional governance of the health systemin relation to management strategies and disputes. METHODOLOGICAL PROCEDURES A qualitative study with health managers from 19 municipalities in the health region of Bahia, Northeastern Brazil. Data were drawn from 17 semi-structured interviews of state, regional, and municipal health policymakers and managers; a focus group; observations of the regional interagency committee; and documents in 2012. The political-institutional and the organizational components were analyzed in the light of dialectical hermeneutics. RESULTS The regional interagency committee is the chief regional governance strategy/component and functions as a strategic tool for strengthening governance. It brings together a diversity of members responsible for decision making in the healthcare territories, who need to negotiate the allocation of funding and the distribution of facilities for common use in the region. The high turnover of health secretaries, their lack of autonomy from the local executive decisions, inadequate technical training to exercise their function, and the influence of party politics on decision making stand as obstacles to the regional interagency committee’s permeability to social demands. Funding is insufficient to enable the fulfillment of the officially integrated agreed-upon program or to boost public supply by the system, requiring that public managers procure services from the private market at values higher than the national health service price schedule (Brazilian Unified Health System Table. The study determined that “facilitators” under contract to health departments accelerated access to specialized (diagnostic, therapeutic and/or surgical services in other municipalities by direct payment to physicians for procedure costs already covered by the Brazilian Unified Health System. CONCLUSIONS The characteristics identified a regionalized system with a conflictive pattern of

  1. Regional governance: strategies and disputes in health region management

    Science.gov (United States)

    dos Santos, Adriano Maia; Giovanella, Ligia

    2014-01-01

    OBJECTIVE To analyze the regional governance of the health systemin relation to management strategies and disputes. METHODOLOGICAL PROCEDURES A qualitative study with health managers from 19 municipalities in the health region of Bahia, Northeastern Brazil. Data were drawn from 17 semi-structured interviews of state, regional, and municipal health policymakers and managers; a focus group; observations of the regional interagency committee; and documents in 2012. The political-institutional and the organizational components were analyzed in the light of dialectical hermeneutics. RESULTS The regional interagency committee is the chief regional governance strategy/component and functions as a strategic tool for strengthening governance. It brings together a diversity of members responsible for decision making in the healthcare territories, who need to negotiate the allocation of funding and the distribution of facilities for common use in the region. The high turnover of health secretaries, their lack of autonomy from the local executive decisions, inadequate technical training to exercise their function, and the influence of party politics on decision making stand as obstacles to the regional interagency committee’s permeability to social demands. Funding is insufficient to enable the fulfillment of the officially integrated agreed-upon program or to boost public supply by the system, requiring that public managers procure services from the private market at values higher than the national health service price schedule (Brazilian Unified Health System Table). The study determined that “facilitators” under contract to health departments accelerated access to specialized (diagnostic, therapeutic and/or surgical) services in other municipalities by direct payment to physicians for procedure costs already covered by the Brazilian Unified Health System. CONCLUSIONS The characteristics identified a regionalized system with a conflictive pattern of governance and

  2. REGIONAL DEVELOPMENT'S PERSPECTIVES IN EUROPE

    Directory of Open Access Journals (Sweden)

    Mihai, OPRESCU

    2014-11-01

    Full Text Available Increasing regions' importance in Europe represents a striking phenomenon in the last years. Regional development is an evolving process that aims at reducing socio-economic disparities at the level of a certain territory, while boosting its potential. One of the European Union objectives is to sustain balanced and sustainable economic and social progress, strengthening economic and social cohesion of member countries. An important role in this process is played by the regions, which are considered closer to the citizens. In European states, the forms of regionalization correspond to different administrative and political realities existing in those countries. Models applied varies by the powers given to regional authorities, involved institutions, the degree of decentralization, being several types of regionalization.

  3. Status of Goos Regional Alliances

    OpenAIRE

    2002-01-01

    Creation of regional GOOS bodies has substantially increased the number of Member States engaged in GOOS. This is happening through three complementary mechanisms: (i) creation of subsidiary bodies within the intergovernmental structure of the IOC (NEAR-GOOS, IOCARIBE-GOOS, Black Sea GOOS, WIOMAP); (ii) creation of regional groups within alternative intergovernmental structures operating on behalf of IOC (PacificGOOS in SOPAC); and (iii) creation of regional associations (EuroG...

  4. Regional approaches to hospital preparedness.

    Science.gov (United States)

    Maldin, Beth; Lam, Clarence; Franco, Crystal; Press, David; Waldhorn, Richard; Toner, Eric; O'Toole, Tara; Inglesby, Thomas V

    2007-03-01

    This article describes issues related to the engagement of hospitals and other community partners in a coordinated regional healthcare preparedness and response effort. The report is based on interviews with public health and hospital representatives from 13 regions or states across the country. It aims to identify key ingredients for building successful regional partnerships for healthcare preparedness as well as critical challenges and policy and practical recommendations for their development and sustainability.

  5. Regional Inflation and Financial Dollarization

    OpenAIRE

    Brown, Martin; de Haas, Ralph; Sokolov, Vladimir

    2013-01-01

    We exploit variation in consumer price inflation across 71 Russian regions to examine the relationship between the perceived stability of the domestic currency and financial dollarization. Our results show that regions with higher inflation experience an increase in deposit dollarization and a decrease in the dollarization of loans to households and firms in non-tradable sectors. The negative impact of inflation on credit dollarization is weaker in regions with less integrated banking markets...

  6. Institution Building for African Regionalism

    OpenAIRE

    Khadiagala, Gilbert M.

    2011-01-01

    Since the 1960s, African states have embraced regional integration as a vital mechanism for political cooperation and for pooling resources to overcome problems of small and fragmented economies. In building meaningful institutions for regionalism, however, Africans have faced the challenges of reconciling the diversities of culture, geography, and politics. As a result, African regional institutions are characterized by multiple and competing mandates and weak institutionalization. This stud...

  7. Regional Stability & Lessons Learned in Regional Peace Building

    DEFF Research Database (Denmark)

    Vestenskov, David; Johnsen, Anton Asklund

    , and many of them were addressed at the conference: The withdrawal of international military forces, the Taliban, India/Pakistan relations, the need for regionalism, China’s role, EU and ASEAN as models for inspiration and regional perspectives from Iran and Afghanistan were all part of the talks...

  8. METHODOLOGY OF REGIONAL ECONOMY OPTIMIZING

    Directory of Open Access Journals (Sweden)

    K. Ainabek

    2016-02-01

    Full Text Available Sustainable development of the national economy directly depends on the optimality of its composite parts. This can be addressed through the appropriate balance of the sectors and the regions, the inner parts of their structure based on a common criterion of the evaluation of cost variables characterizing the parameters of the functioning of the regional actors. This paper shows theoretical principles and economic-mathematical model of optimization of the regional economy and the relations between the center and the regions based on the author’s approach.

  9. Regions and the Territorial Cohesion

    Directory of Open Access Journals (Sweden)

    Ioan Ianos

    2013-08-01

    Full Text Available Territorial cohesion is an important target of European Union, constantly promoted by its institutions and their representatives. In the context of the Europe 2020 strategy, one of the most important support documents, the region represents a very important issue, being considered to be the key to its successfulness. The region is seen as a support for the smart growth and all the operational policy concepts try to make use of the spatial potential, by taking better account of the territorial specificities. Two main questions play attention: the need to transform the present-day developmental regions into administrative ones is a priority? What kind of regionalization it must to be promoted? Correlating these issues with already defined territorial cohesion, the administrative region is a real tool for the future territorial development. The experience of the last 14 years asks urgently the building of a new territorial administrative reform, giving competences to regions. For instant, each development region is a construction resulted from a free association of the counties. Their role in the regional development is much reduced one, because their regional councils are not elected; decisions taken at this level are consultative for the social, economical, cultural or political actors.

  10. Hawaii Regional Sediment Management: Regional Sediment Budget for the Poipu Region of Kauai, HI

    Science.gov (United States)

    2013-06-01

    represent regular maintenance activities such as dredging, bypassing, or nourishment . The region addressed in this study has experienced one or more of these...Hawaii Coastal Geology Group US United States USACE US Army Corps of Engineers USGS US Geological Survey WIS Wave Information Study ERDC/CHL...This Coastal and Hydraulics Engineering Technical Note (CHETN) reviews the development of a regional sediment budget for the Poipu Region on the

  11. PLANNING DESIGN OFCOMPUTER REGION NETWORK

    Institute of Scientific and Technical Information of China (English)

    SongWei; GaoZhuang; GaoSong

    1996-01-01

    In this paper, authors describe the characters and the present situation of computer region network. Moreover, authors put forward the method to plan and design ration ally the computer regional network system depending on the actual situation, in order to share the resources of software and hardware resources of the network in best way and get higher efficiency of information management.

  12. Adoption Resource Directory: Region X.

    Science.gov (United States)

    1983

    State, regional, and national adoption resources are described in this directory for residents of Region X states (Alaska, Idaho, Oregon, and Washington). Emphasizing the adoption of children with special needs, the directory gives organizational contacts for parents in various stages of the adoption process and mentions resources for social…

  13. Regional Inflation and Financial Dollarization

    NARCIS (Netherlands)

    Brown, M.; de Haas, R.; Sokolov, V.

    2013-01-01

    Abstract: We exploit variation in consumer price inflation across 71 Russian regions to examine the relationship between the perceived stability of the local currency and financial dollarization. Our results show that regions with higher inflation experience an increase in the dollarization of house

  14. The political economy of regionalism

    NARCIS (Netherlands)

    S. Goyal (Sanjeev); K. Staal (Klaas)

    2004-01-01

    textabstractWe examine the incentives of regions to unite and separate. Separation allows for greater influence over the nature of political decision making while unification allows regions to exploit economies of scale in the provision of government. Our paper explores the influence of size, locati

  15. Analysis of regional climate strategies in the Barents region

    Energy Technology Data Exchange (ETDEWEB)

    Himanen, S.; Inkeroeinen, J.; Latola, K.; Vaisanen, T.; Alasaarela, E.

    2012-11-15

    Climate change is a global phenomenon with especially harsh effects on the Arctic and northern regions. The Arctic's average temperature has risen at almost twice the rate as elsewhere in the past few decades. Since 1966, the Arctic land area covered by snow in early summer has shrunk by almost a fifth. The Barents Region consists of the northern parts of Norway, Sweden, Finland and Russia (i.e. the European part of Russia). Climate change will cause serious impacts in the Barents Region because of its higher density of population living under harsh climatic conditions, thus setting it apart from other Arctic areas. In many cases, economic activities, like tourism, rely on certain weather conditions. For this reason, climate change and adaptation to it is of special urgency for the region. Regional climate change strategies are important tools for addressing mitigation and adaptation to climate change as they can be used to consolidate the efforts of different stakeholders of the public and private sectors. Regional strategies can be important factors in achieving the national and international goals. The study evaluated how the national climate change goals were implemented in the regional and local strategies and programmes in northern Finland. The specific goal was to describe the processes by which the regional strategies were prepared and implemented, and how the work was expanded to include the whole of northern Finland. Finally, the Finnish preparatory processes were compared to case examples of processes for preparing climate change strategies elsewhere in the Barents Region. This analysis provides examples of good practices in preparing a climate change strategy and implementing it. (orig.)

  16. COMBINATION OF GOALS STRATEGY REGION

    Directory of Open Access Journals (Sweden)

    Denys Yu. Lapigin

    2015-01-01

    Full Text Available Currently the tools to identify strategicallyimportant objectives of regional development is not enough to build a developmentperspective, relying on something special,what distinguishes each region from therest. The article discusses approaches to the formation of the regional developmentstrategy, which is based on goals set by the results of the analysis of the main factors inthe development of the region. The study is based on the methodology of systems theoryand methods of strategic management. The most important results should include tools tobuild the tree of strategic objectives resultingfrom the implementation of the algorithm forconstructing planes of analysis and development of the region. The results can be used to develop a strategy for the developmentof socio-economic systems of various typesand forms.

  17. regional. Un análisis para las regiones europeas

    Directory of Open Access Journals (Sweden)

    José A. Camacho

    2005-01-01

    Full Text Available Pese a encontrarnos inmersos en una economía globalizada, la innovación es en muchos aspectos un fenómeno esencialmente regional, donde los servicios (ignorados hasta ahora, y en particular aquellos denominados como servicios intensivos conocimiento (SIC, adquieren un papel central, gracias a su capacidad para transmitir conocimiento, en especial de tipo tácito. El objetivo de este trabajo es realizar una primera aproximación empírica a los efectos asociados a la localización de SIC, tomando 107 regiones europeas. Los resultados obtenidos apoyan la hipótesis de que los SIC ejercen un impacto positivo sobre el desempeño innovador regional, impacto que podría desbordarse hacia las regiones vecinas.

  18. Planned regional development: the creation of learning regions

    NARCIS (Netherlands)

    Kersten, P.H.; Kranendonk, R.P.; Laurentzen, L.

    2008-01-01

    In this paper, new ways of working in business and management, regional development and governmental steering (phases, levels, changes and growth perspectives) will be identified and described using both theoretical and empirical evidence from the case of Greenport Venlo.

  19. Regiune, regional, regionalizare şi reprezentare politică (Region, regional, regionalisation and political representation

    Directory of Open Access Journals (Sweden)

    Dragoş-Andrei PREUTESCU

    2014-02-01

    Full Text Available Romania is at a crossroad to shape and redefine its regional territorial-administrative design as an European Union member. A regionalization process is at a controversial point in this moment and is necessarry to see if this process could have a significant impact to develop and consolidate democracy in this country. Following the main aspects of regionalism, regionalization and political discourses amoung politicians, the purpose of this article is to see if their is any kind of connection between regionalization process and the involvement of the citizens in public decisions. More so, the new institutional design could consolidate democracy and increase the public responsabilities from both sides: decision makers and citizens. Doing so could lead to better public policies in accordance with needs, expectations and desires of the citizens. In this way, what is called as „regional identity”, or„ local identity” could have more significance in the public imagination and consciousness. Furthermore the so called „movement” for better and developed communities is being shaped in this direction by political discourses and political ideas. What can create better conditions for shaping these democratic options is to have an important political and civic culture once the regionalization process is implemented. The progression of economy, tourism, political representation, culture, and institutions is a consequence of how much these elements are compatible in the regionalization process.

  20. Regional strategies for global leadership.

    Science.gov (United States)

    Ghemawat, Pankaj

    2005-12-01

    The leaders of such global powerhouses as GE, Wal-Mart, and Toyota seem to have grasped two crucial truths: First, far from becoming submerged by the rising tide of globalization, geographic and other regional distinctions may in fact be increasing in importance. Second, regionally focused strategies, used in conjunction with local and global initiatives, can significantly boost a company's performance. The business and economic data reveal a highly regionalized world. For example, trade within regions, rather than across them, drove the surge of international commerce in the second half of the twentieth century. Regionalization is also apparent in foreign direct investment, companies' international sales, and competition among the world's largest multinationals. Harvard Business School Professor Pankaj Ghemawat says that the most successful companies employ five types of regional strategies in addition to--or even instead of--global ones: home base, portfolio, hub, platform, and mandate. Some companies adopt the strategies in sequence, but the most nimble switch from one to another and combine approaches as their markets and businesses evolve. At Toyota, for example, exports from the home base continue to be substantial even as the company builds up an international manufacturing presence. And as Toyota achieves economies of scale and scope with a strong network of hubs, the company also pursues economies of specialization through interregional mandates. Embracing regional strategies requires flexibility and creativity. A company must decide what constitutes a region, choose the most appropriate strategies, and mesh those strategies with the organization's existing structures. In a world that is neither truly global nor truly local, finding ways of coordinating within and across regions can deliver a powerful competitive advantage.

  1. Regional climate change mitigation analysis

    Energy Technology Data Exchange (ETDEWEB)

    Rowlands, Ian H. [UNEP Collaborating Centre on Energy and Environment, and Univ. of Waterloo (Canada)

    1998-10-01

    The purpose of this paper is to explore some of the key methodological issues that arise from an analysis of regional climate change mitigation options. The rationale for any analysis of regional mitigation activities, emphasising both the theoretical attractiveness and the existing political encouragement and the methodology that has been developed are reviewed. The differences arising from the fact that mitigation analyses have been taken from the level of the national - where the majority of the work has been completed to date - to the level of the international - that is, the `regional` - will be especially highlighted. (EG)

  2. REGIONAL CHARACTERISTICS OF CANADIAN ENGLISH

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Modern English is an international language inthe world.Besides Great Britain,English is spokenas first language in 39 countries.These countries arelocated in different regions with different naturalfeatures,history development and cultural character-istics.Thus,English used in these different regionscarries its own regional character—forming Englishregional varieties.The main English regional varieties are:BritishEnglish,American English,Canadian English andSouth African English.Canada is a rich country inNorth America with its own characteristics,which of

  3. Hydrologic landscape regions of Nevada

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Hydrologic landscape regions group areas according to their similarity in landscape and climate characteristics. These characteristics represent variables assumed to...

  4. Region 7 Title V facilities

    Data.gov (United States)

    U.S. Environmental Protection Agency — This web map shows the Region 7 Title V facilities (Clean Air Act major sources), any Class I areas within 300 km of R7 States, and any Tribal areas within 50 miles...

  5. The Landform Regions of Iowa

    Data.gov (United States)

    Iowa State University GIS Support and Research Facility — A landscape is a collection of land shapes or land forms. Landform regions are a grouping of individual landscape features that have a common geomophology. In Iowa,...

  6. Conformity Adequacy Review: Region 9

    Science.gov (United States)

    Resources are for air quality and transportation government and community leaders. Information on the adequacy/inadequacy of state implementation plans (SIPs) in EPA Region 9 (AZ, CA, HI, NV, AS, GU) is provided here.

  7. Groundwater Vulnerability Regions of Iowa

    Data.gov (United States)

    Iowa State University GIS Support and Research Facility — The regions onThis map represent areas with similar hydrogeologic characteristics thought to represent similar potentials for contamination of groundwater and/or...

  8. Census Bureau Regional Office Boundaries

    Data.gov (United States)

    US Census Bureau, Department of Commerce — The Census Bureau has six regional offices to facilitate data collection, data dissemination and geographic operations within their boundary. The surveys these...

  9. Regions compete for French synchrotron

    CERN Multimedia

    2000-01-01

    Ten regions in France have placed bids to host the planned national synchrotron Soleil. Leading contenders include a joint bid from Ile-de-France and Essonne for Orsay, offering FF 1 billion towards the construction costs (2 paragraphs).

  10. [Regional anesthesia and cancer immunology].

    Science.gov (United States)

    Igarashi, Toru; Mori, Katsuya; Inoue, Kei; Suzuki, Takeshi; Morisaki, Hiroshi

    2013-09-01

    Regional anesthesia has been widely applied as an excellent method for perioperative analgesia. Recent studies suggested that regional anesthesia is a promising approach to minimize the risk of surgical site infection and postoperative cancer recurrence, subsequently providing the benefits to the long-term outcome. In particular, it is of great interest that regional anesthesia might be able to reduce cancer recurrence. In cancer patients, innate immunity against cancer could be depressed, resulting in the predisposition to evoke metastasis. Besides, during the perioperative periods, tumor immunity is significantly depressed due to surgical pain, activation of sympathetic nervous system, inflammatory responses, and others. In this review article, we discuss the tumor immunity during the perioperative period, with focus on the alterations of tumor immunity and regional anesthesia.

  11. Regional transport sector mitigation options

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Peter [EECG Consultants, Gaborone (Botswana)

    1998-10-01

    The rationale for conducting climate change mitigation studies in the transport sector is on the premise that: The transport sector is the second largest consumer of fossil fuels in the region; The regional transport sector is an area with high opportunity for infrastructural development under UNFCCC financial mechanism; The regional transport sector is crucial in the SADC region for trade and coupled with the Trade Protocol will play a major role in development hence the need to make it efficient in terms of energy demand and provision of services; The sector offers many mitigation options but with a challenge to evaluate their energy saving and GHG saving potential and yet there is need to quantify possible emission reduction for possible future emission trading. This is also a sector with potential to qualify for financing through Clean Development Mechanism (CDM) recently stipulated in the Kyoto Protocol. (au)

  12. Euroregions in the Podrinje region

    Directory of Open Access Journals (Sweden)

    Tošić Branka

    2008-01-01

    Full Text Available Regionalization is a contemporary conception of a development within a society. We are interested in cooperation at regions whose area is intersected by the national border. One of those borders is the Drina River that in different periods had changeable role of connecting or separating. In this way we want to stress the necessity of making a stronger, functional connection and developing of overall process of integration in the Podrinje region, on a land between the two republics-Serbia and the Republic of Srpska, because we think that this area has the most reasons for that, but so far it has been done very little. The first attempts to form Euroregions in Lower Podrinje region have already started, but it lacks many activities for this area to receive a cross-border role that it deserves.

  13. Vietnam and the regional crisis

    DEFF Research Database (Denmark)

    Masina, Pietro Paolo

    2002-01-01

    inflows, should be understood in the broader frame of regional contingencies, and not be simplistically explained as investors' discontent (i.e., in order to pressure Vietnamese authorities into implementing a more orthodox neo-liberal agenda). And in a post-crisis regional economic reorganisation......The paper explores the linkages between Vietnam and the regional economy before and after the East Asian financial crisis of 1997 and 1998, indicating that the country is more closely associated with the region than normally assumed. Thus, key indicators, such as falling foreign direct investment...... (including a restructuring of the so-called 'flying geese' pattern) Vietnam's chances of benefitting from closer integration in the world economy will depend on a clearcut industrial and trade policy, rather than on embarking a so-called 'neutral trade regime'. Preliminary evidence seems to indicate...

  14. Region 9 NPL Sites - 2014

    Data.gov (United States)

    U.S. Environmental Protection Agency — NPL site point locations for the US EPA Region 9. NPL (National Priorities List) sites are hazardous waste sites that are eligible for extensive long-term cleanup...

  15. US Forest Service Regional Boundaries

    Data.gov (United States)

    US Forest Service, Department of Agriculture — A map service on the www depicting all the National Forest System lands administered by a Region. The area encompasses private lands, other governmental agency...

  16. Regions.

    Science.gov (United States)

    Moulin-Acevedo, Madeleine; And Others

    1993-01-01

    Includes "From School to Jobs: Africa's Dilemma" (Moulin-Acevedo); "Helping Change in Eastern Europe"; "Recognizing the Dignity of Indigenous Peoples"; "An Employment Plan for Pakistan"; and "Around the Continents." (JOW)

  17. ESPC Regional Arctic Prediction System

    Science.gov (United States)

    2014-09-30

    the Navy the capability to conduct short-term (1 week) to extended (2 weeks) coupled weather forecasts for the Arctic region. APPROACH To...sensitivity of the Arctic weather forecast to key numerical parameters; and 5) conduct extensive validation and verification of the coupled system and...SEP 2014 2. REPORT TYPE 3. DATES COVERED 00-00-2014 to 00-00-2014 4. TITLE AND SUBTITLE ESPC Regional Arctic Prediction System 5a. CONTRACT

  18. Finding Distant Galactic HII Regions

    CERN Document Server

    Anderson, L D; Johnstone, B M; Bania, T M; Balser, Dana S; Wenger, Trey V; Cunningham, V

    2015-01-01

    The WISE Catalog of Galactic HII Regions contains $\\sim2000$ HII region candidates lacking ionized gas spectroscopic observations. All candidates have the characteristic HII region mid-infrared morphology of WISE $12\\,\\,\\mu\\,m$ emission surrounding $22\\,\\mu\\,m$ emission, and additionally have detected radio continuum emission. We here report Green Bank Telescope (GBT) hydrogen radio recombination line (RRL) and radio continuum detections at X-band (9GHz; 3cm) of 302 WISE HII region candidates (out of 324 targets observed) in the zone $225^{\\circ} > l > -20^{\\circ}$, $|b| \\le 6^{\\circ}$. Here we extend the sky coverage of our HII region Discovery Survey (HRDS), which now contains nearly 800 HII regions distributed across the entire northern sky. We provide LSR velocities for the 302 detections and kinematic distances for 131 of these. Of the 302 new detections, five have ($l, b, v$) coordinates consistent with the Outer Scutum-Centaurus Arm (OSC), the most distant molecular spiral arm of the Milky Way. Due to ...

  19. Solar transition region above sunspots

    CERN Document Server

    Tian, H; Teriaca, L; Landi, E; Marsch, E

    2009-01-01

    We study the TR properties above sunspots and the surrounding plage regions, by analyzing several sunspot spectra obtained by SUMER in March 1999 and November 2006. We compare the SUMER spectra observed in the umbra, penumbra, plage, and sunspot plume regions. The Lyman line profiles averaged in each region are presented. For the sunspot observed in 2006, the electron densities, DEM, and filling factors of the TR plasma in the four regions are also investigated. The self-reversals of the Lyman line profiles are almost absent in umbral regions at different locations (heliocentric angle up to $49^\\circ$) on the solar disk. In the sunspot plume, the Lyman lines are also not reversed, whilst the lower Lyman line profiles observed in the plage region are obviously reversed. The TR densities of the umbra and plume are similar and one order of magnitude lower than those of the plage and penumbra. The DEM curve of the sunspot plume exhibits a peak centered around $\\log(T/\\rm{K})\\sim5.45$, which exceeds the DEM of oth...

  20. Social attitudes and regional inequalities

    Directory of Open Access Journals (Sweden)

    Vinko Muštra

    2014-03-01

    Full Text Available As the budgets of the European Union (EU become tighter, the questions about the effectiveness of EU’s expenditure, especially the most important parts such as Cohesion policy, are hotly debated. The aim of this paper is to examine whether the presence of social attitudes may influence the effectiveness of EU budget expenditure, measured by the level of regional inequalities. The analysis starts by focusing on individuals’ attitudes towards income from their own effort and income which is derived from other people’s effort, having in mind that individual actions depend on their attitudes. The next step establishes the link between the income from other people’s effort with the re-distributive dimension of the EU budget, considering that different attitudes among individuals in the EU could lead to significant differences in effectiveness of this redistributive policy among European regions and, consequently, diverse regional inequalities. Empirical research uses data for 27 EU countries observed over two waves of European Value Surveys: 1999-2000 (Wave 1 and 2008–2009 (Wave 2. The results indicate a significant role of social attitudes for regional inequalities, which raises the question of the appropriateness of simplification and uniform regional policy instruments in solving EU regional problems.

  1. Promotion and regional development. Implementation of regional productive development agencies. The case of Maule region, Chile

    Directory of Open Access Journals (Sweden)

    Enrique Yamil Alul González

    2010-12-01

    Full Text Available The Regional Productive Development Agencies implemented in Chile in 2006, were developed as a way to answer the longing desire to territorially decentralize, and that the own Regions be whom define their future. The Agencies have the responsibility to develop innovation and productive development Agendas in participative processes, which means with public, academic and private actors. Also, the Agencies have the mission to implement Competitive Improvement Plans-PMC (clusters in prioritized economic sectors by the own region. These PMC are leaded by private actors in each sector.

  2. Traditional and regional food in Poland

    OpenAIRE

    Gulbicka, Bożena

    2014-01-01

    Regional and traditional products in the European Union - basic legal regulations. Traditional and regional products in the Polish legislation. National and regional food quality schemes. Quality and safety or traditional and regional food. Polish traditional and regional products registered with the European Union and their characteristics. Opportunities for and barriers to the development of the market of traditional and regional products in Poland.

  3. REGIONAL ECONOMIC GROWTH THROUGH TOURISM. THE CASE OF REGION WEST

    Directory of Open Access Journals (Sweden)

    Dragoi Ionut

    2010-12-01

    Full Text Available The regional development should aim to correlate and integrate tourism among the other integrative parts of the regional and local development, taking into account the fact that a region’s prosperity as effect of tourism development may be shown clearly in several stages: on the spot (as a result of direct consumption of the tourist product, on short term (through continuous absorption of the work-force and encouraging the welcoming commerce and in the long run (concentrating capital for investment in the general infrastructure and the one of tourism, in structures of reception for tourism and in the development of urban services. The analysis of intra-regional disparities as part of the economic growth at the level of Region West starts off with the idea that each component county has a different landscape, which favored or inhibited their economic growth; in the same time, each component county has its own specific, which can be promoted through tourism, inducing in time a regional income, and respectively, a social-economic and cultural growth of less developed areas.

  4. Regional sinkhole susceptibility maps: The Latium Region case (central Italy)

    Science.gov (United States)

    La Vigna, F.; Teoli, P.; Mazza, R.; Leoni, G.; Capelli, G.

    2012-04-01

    Several and frequent studies were internationally presented about landslide susceptibility, meanwhile in literature is missing a broad diffusion of studies regarding sinkhole susceptibility. That's why sinkhole recurrence depends on several geological conditions related to specific geological and hydrogeological context (sinkhole prone area) that vary case by case. Notwithstanding this regionalization problem of sinkhole recurrence, in the central Appenine sedimentary basins (Italy) a certain number of geological, geomorphologic and hydrogeological conditions (sinkhole predisposing issues) can be considered in common between the surveyed sinkholes. Eventually this could be compared with similar geological conditions and sinkhole occurrence in the rest of Italy or in other countries. In this case study is presented a probabilistic approach regarding the Latium Region deriving from the comparison between the regional sinkhole inventory realized during a precedent project and the dataset of the new Hydrogeological Map of Latium Region (scale 1:100.000). Indexed elements, chosen because associated to the majority of sinkhole phenomena, are: outcropping lithologies, water table depth, main faults (even if buried), hydrothermal springs, land use and the epicentres of recent earthquakes. These indexed elements were weighted and combined in a matrix which preliminary result is the sinkhole susceptibility map of Latium Region. When definitively validated, this approach could be suitable for local authorities to planning more targeted studies in major hazard areas.

  5. Users guide to REGIONAL-1: a regional assessment model

    Energy Technology Data Exchange (ETDEWEB)

    Davis, W.E.; Eadie, W.J.; Powell, D.C.

    1979-09-01

    A guide was prepared to allow a user to run the PNL long-range transport model, REGIONAL 1. REGIONAL 1 is a computer model set up to run atmospheric assessments on a regional basis. The model has the capability of being run in three modes for a single time period. The three modes are: (1) no deposition, (2) dry deposition, (3) wet and dry deposition. The guide provides the physical and mathematical basis used in the model for calculating transport, diffusion, and deposition for all three modes. Also the guide includes a program listing with an explanation of the listings and an example in the form of a short-term assessment for 48 hours. The purpose of the example is to allow a person who has past experience with programming and meteorology to operate the assessment model and compare his results with the guide results. This comparison will assure the user that the program is operating in a proper fashion.

  6. NWRS Region 7 Inventory & Monitoring Regional Annual Report, FY2011 : Alaska Region FY 2011 Annual Report

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This annual report for Region 7 discusses the goals and objectives of the Inventory and Monitoring (I&M) program for fiscal year 2011. The introduction discusses...

  7. Regional Community and International Relations: the Volgograd Region Experience

    Directory of Open Access Journals (Sweden)

    Danakari Richard A.

    2016-06-01

    Full Text Available The article deals with the complex and controversial problems of the new regional communities’ formation and the impact of the interethnic relations sphere on them. The author notes that the processes of interaction between representatives of different cultures and civilizations, ethnic groups and religions have become increasingly controversial and tense in the context of continuous social dynamics. Similarly to the Russian society as a whole, regional communities are in a state of transitivity. They get transformed, they acquire new qualities such as multicasting and heterogeneity, multi-ethnicity and multi-confessionalism, fragmentarity and multiculturality. This fact increases the risks and uncertainties, problematizes future prospects. National non-governmental organizations are increasingly positioning themselves as civil society institutions at the present stage of social development at the regional level. They perform a difficult dual task: on the one hand, they ensure the preservation and development of history, native language, culture, ethnic traditions, and on the other hand, they work on the integration, on the common identity and the Russian nation formation. On the territory of the Volgograd region, largely due to the active cooperation of regional authorities and local authorities with national public associations, international and inter-confessional relations are stable. The basis of such activity is respect for history, native language, culture, tradition, religion, national dignity of all people in the region, regardless of their belonging to a certain ethnic group or religion. Over two decades of accumulated considerable experience of joint inter-ethnic dialogue and cooperation, provided tolerance and peace, harmony and mutual understanding between people of different ethnicities and religions in the country.

  8. Regional Transmission Projects: Finding Solutions

    Energy Technology Data Exchange (ETDEWEB)

    The Keystone Center

    2005-06-15

    The Keystone Center convened and facilitated a year-long Dialogue on "Regional Transmission Projects: Finding Solutions" to develop recommendations that will help address the difficult and contentious issues related to expansions of regional electric transmission systems that are needed for reliable and economic transmission of power within and across regions. This effort brought together a cross-section of affected stakeholders and thought leaders to address the problem with the collective wisdom of their experience and interests. Transmission owners sat at the table with consumer advocates and environmental organizations. Representatives from regional transmission organizations exchanged ideas with state and federal regulators. Generation developers explored common interests with public power suppliers. Together, the Dialogue participants developed consensus solutions about how to begin unraveling some of the more intractable issues surrounding identification of need, allocation of costs, and reaching consensus on siting issues that can frustrate the development of regional transmission infrastructure. The recommendations fall into three broad categories: 1. Recommendations on appropriate institutional arrangements and processes for achieving regional consensus on the need for new or expanded transmission infrastructure 2. Recommendations on the process for siting of transmission lines 3. Recommendations on the tools needed to support regional planning, cost allocation, and siting efforts. List of Dialogue participants: List of Dialogue Participants: American Electric Power American Transmission Company American Wind Energy Association California ISO Calpine Corporation Cinergy Edison Electric Institute Environmental Defense Federal Energy Regulatory Commission Great River Energy International Transmission Company ISO-New England Iowa Public Utility Board Kanner & Associates Midwest ISO National Association of Regulatory Utility Commissioners National Association

  9. Hawaii Regional Sediment Management: Regional Sediment Budget for the Kekaha Region of Kauai, HI

    Science.gov (United States)

    2013-06-01

    study regions, have been developed by the University of Hawaii Coastal Geology Group (UH CGG) (Fletcher et al. 2012) for the US Geological Survey...maintenance activities such as dredging, bypassing, or nourishment . The region addressed in this study has experienced one or more of these activities... Coastal Geology Group US United States USACE US Army Corps of Engineers USGS US Geological Survey WIS Wave Information Study ERDC/CHL CHETN

  10. Regional Development Disparities In Europe

    Directory of Open Access Journals (Sweden)

    Liliana - Craciun

    2013-08-01

    Full Text Available Regional economic integration provides both, developing countries and the least developed, the ability to enjoy the benefits of a larger „European “market, whether it is their home or their adoptive home. This issue gains a larger dimension in the context of economic crisis and euro zone. The argument for this statement is that regional development disparities may negatively affect economic cohesion from European space. There were identified two obstacles of the efficient use of European resources. The first one is the geographic barrier: the inability to make labor division due to barrier restrictions. The second one is the lack of an entrepreneurial culture. The entrepreneurial culture provides the flexibility of economy - in particular, the structural flexibility to cope with changes in the division of labor. These disparities can be gradual changed, and they are primarily result from autonomous technological innovations made ​​in response to depletion of resources or affected environment. Analyzing regional development disparities there were applied well known research methods: analytical and statistical method. The analysis consists on selecting and describing a set of indicators “measures” for regional competitiveness, able to show the situation of the region in metric terms, but also from economic point of view.

  11. Regional welfare disparities and regional economic growth in Vietnam

    NARCIS (Netherlands)

    Nguyen Huy Hoang,

    2009-01-01

    This study had multiple objectives. First, it aimed at examining regional inequality in several welfare variables, such as health and education during the 1998-2004 period, and per capita income over time from 1990 to 2006. The first objective of the study was to gain a better understanding about

  12. Vital coalitions, vital regions; cooperation in sustainable, regional development

    NARCIS (Netherlands)

    Horlings, L.G.

    2010-01-01

    Many rural regions in Europe are undergoing a dynamic transition, driven by forces of urbanisation and agricultural development, new patterns of production and consumption, and new societal demands. However, while the historical rural-urban divide is eroding, rural landscapes are becoming more impor

  13. Regional Cooperation to Strengthen Safeguards

    Energy Technology Data Exchange (ETDEWEB)

    Minnini, Margot; Elkhamri, Oksana O.

    2016-06-06

    President Obama’s decision over four years ago to ”pivot” toward Asia represented an important strategic shift in American foreign policy and a rebalancing of U.S. economic and security engagement in the Asia-Pacific countries. The United States has since supported a variety of regional initiatives aimed at promoting nuclear security and safeguards. When a new regional organization, the Asia-Pacific Safeguards Network (APSN) was established in 2010, DOE/NNSA became an early member and enthusiastic advocate. Launched on the initiative of Australia, Japan, Republic of Korea, and Indonesia, the APSN aims to strengthen the quality and effectiveness of safeguards implementation in the Asia-Pacific region.

  14. Regional sea level change in the Thailand-Indonesia region

    Science.gov (United States)

    Fenoglio-Marc, L.; Becker, M. H.; Buchhaupt, C.

    2013-12-01

    It is expected that the regional sea level rise will strongly affect particular regions with direct impacts including submergence of coastal zones, rising water tables and salt intrusion into groundwaters. It can possibly also exacerbate other factors as floodings, associated to storms and hurricanes, as well as ground subsidence of anthropogenic nature. The Thailand-Vietnam-Indonesian region is one of those zones. On land, the Chao-Praya and Mekong Delta are fertile alluvial zones. The potential for sea level increases and extreme floodings due to global warming makes the Deltas a place where local, regional, and global environmental changes are converging. We investigate the relative roles of regional and global mechanisms resulting in multidecadal variations and inflections in the rate of sea level change. Altimetry and GRACE data are used to investigate the variation of land floodings. The land surface water extent is evaluated at 25 km sampling intervals over fifteen years (1993-2007) using a multisatellite methodology which captures the extent of episodic and seasonal inundations, wetlands, rivers, lakes, and irrigated agriculture, using passive and active (microwaves and visible observations. The regional sea level change is analysed during the period 1993-2012 using satellite altimetry, wind and ocean model data, tide gauge data and GPS. The rates of absolute eustatic sea level rise derived from satellite altimetry through 19-year long precise altimeter observations are in average higher than the global mean rate. Several tide gauge records indicate an even higher sea level rise relative to land. We show that the sea level change is closely linked to the ENSO mode of variability and strongly affected by changes in wind forcing and ocean circulation. We have determined the vertical crustal motion at a given tide gauge location by differencing the tide gauge sea level time-series with an equivalent time-series derived from satellite altimetry and by computing

  15. Connectivity graphs of uncertainty regions

    CERN Document Server

    Chambers, Erin; Lenchner, Jonathan; Sember, Jeff; Srinivasan, Venkatesh; Stege, Ulrike; Stolpner, Svetlana; Weibel, Christophe; Whitesides, Sue

    2010-01-01

    We study a generalization of the well known bottleneck spanning tree problem called "Best Case Connectivity with Uncertainty": Given a family of geometric regions, choose one point per region, such that the length of the longest edge in a spanning tree of a disc intersection graph is minimized. We show that this problem is NP-hard even for very simple scenarios such as line segments and squares. We also give exact and approximation algorithms for the case of line segments and unit discs respectively.

  16. Afghanistan's Role in Regional Cooperation

    Institute of Scientific and Technical Information of China (English)

    Hamid Karzai

    2006-01-01

    @@ At the invitation of Cui Liru, President of China Institutes of Contemporary International Relations (CICIR), Mr.Hamid Karzai,President of the Islamic Republic of Afghanistan, paid a visit to CICIR on June 20, 2006.In his speech before an audience of several hundred researchers, President Karzai addressed many issues Afghanistan is now facing, including the achievements of peace and reconstruction, the necessities and ways for Afghanistan to re-join in the regional affairs and the international community, the role of Afghanistan in the regional development and prosperity, and the meaning of a stable Afghanistan to China. Following are the contents of his speech.

  17. Geospatial Absorption and Regional Effects

    Directory of Open Access Journals (Sweden)

    IOAN MAC

    2009-01-01

    Full Text Available The geospatial absorptions are characterized by a specific complexity both in content and in their phenomenological and spatial manifestation fields. Such processes are differentiated according to their specificity to pre-absorption, absorption or post-absorption. The mechanisms that contribute to absorption are extremely numerous: aggregation, extension, diffusion, substitution, resistivity (resilience, stratification, borrowings, etc. Between these mechanisms frequent relations are established determining an amplification of the process and of its regional effects. The installation of the geographic osmosis phenomenon in a given territory (a place for example leads to a homogenization of the geospatial state and to the installation of the regional homogeneity.

  18. China from a regional perspective

    DEFF Research Database (Denmark)

    Schmidt, Johannes Dragsbæk

    The paper explains the political economic background for China's insertion to the world system. It furthermore expands on a critical perspective on China's soft power strategy. It goes on to discuss China's foreign policy strategy towards Southeast Asia and China's rivalry with the US in the region....

  19. Education and Italian Regional Development

    Science.gov (United States)

    Di Liberto, Adriana

    2008-01-01

    In this paper, we study the connection between growth and human capital in a convergence regression for the panel of Italian regions. We include measures of average primary, secondary and tertiary education. We find that increased education seems to contribute to growth only in the South. Decomposing total schooling into its three constituent…

  20. Biocapacity optimization in regional planning

    Science.gov (United States)

    Guo, Jianjun; Yue, Dongxia; Li, Kai; Hui, Cang

    2017-01-01

    Ecological overshoot has been accelerating across the globe. Optimizing biocapacity has become a key to resolve the overshoot of ecological demand in regional sustainable development. However, most literature has focused on reducing ecological footprint but ignores the potential of spatial optimization of biocapacity through regional planning of land use. Here we develop a spatial probability model and present four scenarios for optimizing biocapacity of a river basin in Northwest China. The potential of enhanced biocapacity and its effects on ecological overshoot and water consumption in the region were explored. Two scenarios with no restrictions on croplands and water use reduced the overshoot by 29 to 53%, and another two scenarios which do not allow croplands and water use to increase worsened the overshoot by 11 to 15%. More spatially flexible transition rules of land use led to higher magnitude of change after optimization. However, biocapacity optimization required a large amount of additional water resources, casting considerable pressure on the already water-scarce socio-ecological system. Our results highlight the potential for policy makers to manage/optimize regional land use which addresses ecological overshoot. Investigation on the feasibility of such spatial optimization complies with the forward-looking policies for sustainable development and deserves further attention.

  1. Renewable Energy in European Regions

    NARCIS (Netherlands)

    Krozer, Y.

    2012-01-01

    The regional dynamics of energy innovation, in particular the shift from fossil fuels to renewable energy in the EU, is discussed within the framework of neo-Schumpeterian theory. The EU’s 4.2% average annual growth in renewable energy production in the last decade has been accompanied by diverging

  2. Metropolitan region of Copenhagen, Denmark

    DEFF Research Database (Denmark)

    Fertner, Christian

    the boundaries of the city further out. With the opening of the suburban train lines in 1934, the until today known finger structure was consolidated. A regional strategy, the “Fingerplan”, incorporating this structure was elaborated in 1947, but individual motorisation eroded some of the principles. Still...

  3. Snakes of the Guianan region

    NARCIS (Netherlands)

    Hoogmoed, M.S.

    1982-01-01

    The study of snaks from the Guianan region got an early start in 1705 when several species were pictured by Merian. As relatively large proportion of the snakes described by Linnaeus originated from Surinam. Interest for and knowledge of this group of animals steadily increased in the 18th and 19th

  4. CHINA SEEKS REGIONAL ENERGY COOPERATION

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    China is seeking to diversify channels for energy cooperation as it faces mounting challenges from surging energy demand, geopolitical risks and price volatility. The endowment and distribution of China's resources does not match the current situation of China's economic development. Those are the opinions aired by officials and experts at an international expo recently held in West China's Xinjiang Uygur Autonomous Region.

  5. Regional Tax Reform Goes National

    Institute of Scientific and Technical Information of China (English)

    LAN XINZHEN

    2011-01-01

    After a year of experimental reform on the resource tax ratio in China's western Xinjiang Uygur Autonomous Region,the State Council announced on September 21 to add the method of levying the resource tax ratio by value to the existing practice of levying the ratio by volume only.It will also change resource tax rates on crude oil and natural gas.

  6. Regional approaches to water pollution

    NARCIS (Netherlands)

    Rijtema, P.E.; Bolt, van der F.J.E.

    1996-01-01

    General aspects of regional modelling of environmental impacts of non-point sources of pollution are discussed. Models range from simple budget to very complex structural models. In groundwater pollution studies structural models are preferable. The choice of the model depends on the aim of the rese

  7. China's Regional Development and Tibet

    Directory of Open Access Journals (Sweden)

    Kunal Mukherjee

    2016-11-01

    Full Text Available Reviewed: China's Regional Development and Tibet By Rongxing Guo. Singapore: Springer, 2016. xxii + 195 pp. Hardcover: US$ 99.99. ISBN 978-981-287-956-1. E-book: US$ 69.99. ISBN 978-981-287-958-5.

  8. Algorithms for Labeling Focus Regions.

    Science.gov (United States)

    Fink, M; Haunert, Jan-Henrik; Schulz, A; Spoerhase, J; Wolff, A

    2012-12-01

    In this paper, we investigate the problem of labeling point sites in focus regions of maps or diagrams. This problem occurs, for example, when the user of a mapping service wants to see the names of restaurants or other POIs in a crowded downtown area but keep the overview over a larger area. Our approach is to place the labels at the boundary of the focus region and connect each site with its label by a linear connection, which is called a leader. In this way, we move labels from the focus region to the less valuable context region surrounding it. In order to make the leader layout well readable, we present algorithms that rule out crossings between leaders and optimize other characteristics such as total leader length and distance between labels. This yields a new variant of the boundary labeling problem, which has been studied in the literature. Other than in traditional boundary labeling, where leaders are usually schematized polylines, we focus on leaders that are either straight-line segments or Bezier curves. Further, we present algorithms that, given the sites, find a position of the focus region that optimizes the above characteristics. We also consider a variant of the problem where we have more sites than space for labels. In this situation, we assume that the sites are prioritized by the user. Alternatively, we take a new facility-location perspective which yields a clustering of the sites. We label one representative of each cluster. If the user wishes, we apply our approach to the sites within a cluster, giving details on demand.

  9. Regional Integration of Renewable Energies; Integracion Regional de energias Renovables

    Energy Technology Data Exchange (ETDEWEB)

    Amador Guerra, J.; Dominguez Bravo, J. [Ciemat.Madrid (Spain)

    2000-07-01

    The aim of this report is to show how Energetic Planning and Territorial Policy should be working together for a better integration of Renewable Energies into Region. This Integration should to contemplate social, economic and environmental aspects of the territory. The report has been classified into 7 items: planning, energetic scenarios, technology transfer for Renewable Energies dissemination, barriers for this dissemination, environmental aspects, European Union Policy and Decision Support Systems (and specially GIS). (Author) 54 refs.

  10. Social Facilitation: the Kaliningrad Region and Russian Mainland Regions

    Directory of Open Access Journals (Sweden)

    Toropov P.

    2015-07-01

    Full Text Available The authors look at the concept of social facilitation as a possible component of Russian social consolidation in the course of social reforms. The article focuses on the results of an empirical study of the levels and characteristics of competence in social facilitation, which is here understood as the ability to apply certain knowledge and systems of skills and as a success rate of actions based on experience in improving the conditions of social development. The study was conducted in the Kaliningrad region in 2012—2013. The data was obtained through a survey of 400 respondents using the authors’ methodology encompassing eight basic elements of competence and a structured interview aimed at a better understanding of the subject of the study and attitudes towards it. The data is compared against the results of a similar study in two regions of central Russia with similar conditions. The results suggest a disharmonic and inconsistent structure of competence in social facilitation, low motivation for social activity in youth, and a narrow range of ideas about possible areas of personal activities in the current conditions. Gender differences are identified in the level and structure of competence. The authors believe that certain differences in competence components identified through mathematical methods are determined by the geographical characteristics of the Kaliningrad region — its exclave nature, a relatively small territory, and proximity to the EU countries. It is stressed that the regional conditions affecting motivation, forms and areas of activities, and structure of experience should be taken into account in

  11. Allegheny County Kane Regional Center Census

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — Total number of residents in each Kane Regional Center facility by race and gender. The Kane Regional Centers are skilled nursing and rehabilitation centers run by...

  12. collision region of the ISR

    CERN Multimedia

    1970-01-01

    This is a collision region from the world’s first proton collider, the Intersecting Storage Rings. The ISR was used at CERN from 1971-84 to study proton-proton collisions at the highest energy then available (60GeV). When operational, ISR collision regions were surrounded by detectors as shown in the photo. In 1972, the surprising discovery of fragments flying out sideways from head-on proton-proton collisions was the first evidence of quark-quark scattering inside the colliding protons . This was similar to Rutherford’s observation in 1911 of alpha particles scattering off the tiny nucleus inside atoms of gold. The ISR beamtubes had to be as empty as outer space, a vacuum 100 000 times better than other CERN machines at the time.

  13. EPA Region 3 Quality Management Plans

    Science.gov (United States)

    Has links to resources that describe the Region's Quality Assurance Program, which is a collection of the Region's ongoing quality assurance (QA) policies, procedures, responsibilities and management systems.

  14. Selected Regional Judicial Officer Cases, 2005 - Present

    Data.gov (United States)

    U.S. Environmental Protection Agency — This dataset contains selected cases involving EPA's Regional Judicial Officers (RJOs) from 2005 to present. EPA's Regional Judicial Officers (RJOs) perform...

  15. Shaping Regional Strategies of Multinational Corporations

    DEFF Research Database (Denmark)

    Gilbert, Dirk Ulrich; Heinicke, Patrick; Rasche, Andreas

    This paper examines the factors that drive the success of multinational corporations (MNCs) in their pursuit of regional strategies. We develop a comprehensive regional success factor model to investigate the effects of regional management autonomy and regional product/service adaption on the reg......This paper examines the factors that drive the success of multinational corporations (MNCs) in their pursuit of regional strategies. We develop a comprehensive regional success factor model to investigate the effects of regional management autonomy and regional product/service adaption...

  16. Southeast Regional Carbon Sequestration Partnership

    Energy Technology Data Exchange (ETDEWEB)

    Kenneth J. Nemeth

    2006-08-30

    The Southeast Regional Carbon Sequestration Partnership's (SECARB) Phase I program focused on promoting the development of a framework and infrastructure necessary for the validation and commercial deployment of carbon sequestration technologies. The SECARB program, and its subsequent phases, directly support the Global Climate Change Initiative's goal of reducing greenhouse gas intensity by 18 percent by the year 2012. Work during the project's two-year period was conducted within a ''Task Responsibility Matrix''. The SECARB team was successful in accomplishing its tasks to define the geographic boundaries of the region; characterize the region; identify and address issues for technology deployment; develop public involvement and education mechanisms; identify the most promising capture, sequestration, and transport options; and prepare action plans for implementation and technology validation activity. Milestones accomplished during Phase I of the project are listed below: (1) Completed preliminary identification of geographic boundaries for the study (FY04, Quarter 1); (2) Completed initial inventory of major sources and sinks for the region (FY04, Quarter 2); (3) Completed initial development of plans for GIS (FY04, Quarter 3); (4) Completed preliminary action plan and assessment for overcoming public perception issues (FY04, Quarter 4); (5) Assessed safety, regulatory and permitting issues (FY05, Quarter 1); (6) Finalized inventory of major sources/sinks and refined GIS algorithms (FY05, Quarter 2); (7) Refined public involvement and education mechanisms in support of technology development options (FY05, Quarter 3); and (8) Identified the most promising capture, sequestration and transport options and prepared action plans (FY05, Quarter 4).

  17. Pollutant transport among California regions

    Science.gov (United States)

    Angevine, Wayne M.; Brioude, Jerome; McKeen, Stuart; Holloway, John S.; Lerner, Brian M.; Goldstein, Allen H.; Guha, Abhinav; Andrews, Arlyn; Nowak, John B.; Evan, Stephanie; Fischer, Marc L.; Gilman, Jessica B.; Bon, Daniel

    2013-06-01

    Several regions within California have significant air quality issues. Transport of pollutants emitted in one region to another region may add to the impact of local emissions. In this work, Lagrangian particle dispersion model simulations show the amounts of tracers that are transported within and among four regions, Southern California, the San Francisco Bay Area, the Central Valley, and the rest of the state. The simulations cover May and June of 2010, the California Research at the Nexus of Air Quality and Climate Change experiment period. Tracers of automobile emissions and one type of agricultural emission are used. Tracer mixing ratios are compared to airborne and ground-based measurements. The age of tracers in each location is also presented. Vertical profiles and diurnal cycles help to clarify the transport process. As is well known, Southern California emissions are transported to the east and affect the desert areas, and Bay Area automobile emissions are an important source of pollutants in the San Joaquin Valley. A novel result is that the Southern California Bight is filled with a mixture of well-aged carbon monoxide tracer from Southern California and the Bay Area. Air over the Bight is also affected by the agricultural emissions represented by the agricultural tracer, dominantly from the Central Valley where its sources are largest. There is no indication of transport from Southern California to the Central Valley. Emissions from the Central Valley do make their way to Southern California, as shown by the agricultural tracer, but automobile emissions from the Valley are insignificant in Southern California.

  18. Regional Tax Reform Goes National

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    China plans to increase resource tax to curb waste,but the plans raise fears of inflation After a year of experimental reform on the resource tax ratio in China’s western Xinjiang Uygur Autonomous Region,the State Council announced on September 21 to add the method of levying the resource tax ratio by value to the existing practice of levying the ratio by volume only.

  19. Dvigateli regional'nogo stroitel'stva. Vlijanie regional'nyh politicheskih organizacij na sotrudnichestvo universitetov v regione Baltijskogo morja [Motors for regional development: impact on regional political organizations on the university cooperation in the Baltic Sea region

    Directory of Open Access Journals (Sweden)

    Ewert Stefan

    2010-01-01

    Full Text Available Educational co-operation is one of the main aspects of the regional political agenda in the Baltic Sea Region. The article analyzes the political impact of the organizations, as perceived by the universities in the region and political decision-makers on national and regional levels. Based on the success of the OECD in becoming an influential actor in educational policies, this article discusses different strategies for the regional political organizations to enhance their influence.

  20. Great Lakes' regional climate regimes

    Science.gov (United States)

    Kravtsov, Sergey; Sugiyama, Noriyuki; Roebber, Paul

    2016-04-01

    We simulate the seasonal cycle of the Great Lakes' water temperature and lake ice using an idealized coupled lake-atmosphere-ice model. Under identical seasonally varying boundary conditions, this model exhibits more than one seasonally varying equilibrium solutions, which we associate with distinct regional climate regimes. Colder/warmer regimes are characterized by abundant/scarce amounts of wintertime ice and cooler/warmer summer temperatures, respectively. These regimes are also evident in the observations of the Great Lakes' climate variability over recent few decades, and are found to be most pronounced for Lake Superior, the deepest of the Great Lakes, consistent with model predictions. Multiple climate regimes of the Great Lakes also play a crucial role in the accelerated warming of the lakes relative to the surrounding land regions in response to larger-scale global warming. We discuss the physical origin and characteristics of multiple climate regimes over the lakes, as well as their implications for a longer-term regional climate variability.

  1. Analgesia regional em cuidados intensivos

    Directory of Open Access Journals (Sweden)

    Luísa Guedes

    2012-10-01

    Full Text Available JUSTIFICATIVAS E OBJETIVOS: A analgesia regional desempenha um papel importante na abordagem multimodal da dor no doente crítico e permite amenizar o desconforto do doente e reduzir os estresses fisiológico e psicológico associados. Ao diminuir as doses de opioides sistêmicos, reduz alguns dos seus efeitos colaterais, como a síndrome de abstinência, possíveis alterações psicológicas e disfunção gastrintestinal. Apesar desses benefícios, seu uso é controverso, uma vez que os doentes em unidades de cuidados intensivos apresentam frequentemente contraindicações, como coagulopatia, instabilidade hemodinâmica e dificuldade na avaliação neurológica e na execução da técnica regional. CONTEÚDO: Os autores apresentam uma revisão sobre analgesia regional em cuidados intensivos, com foco nas principais vantagens e limitações de seu uso no doente crítico, e descrevem as técnicas regionais mais usadas e a sua aplicabilidade nesse contexto.

  2. Regional Filmmaking After Jia Zhangke

    Directory of Open Access Journals (Sweden)

    Mei Yang

    2015-08-01

    Full Text Available This article examines Ying Liang’s films to illustrate Chinese independent filmmakers’ growing propensity for representing regional space through innovative cinematic techniques. As one of the most conspicuous directors onstage in the 2000s, Ying Liang rekindles independent regional film following the legacy of Jia Zhangke’s hometown trilogy. Varying shooting angles and camera positions, his exemplar work of formalistic experimentation, The Other Half (Ling yiban, 2006, presents space as a pivot of relations, within which the position of self and the situation of the local are intertwined. In line with Doreen Massey’s proposition for a spatial turn in theoretical conceptions, I argue that Ying Liang’s exploration of relational cinematic space, between on-screen and offscreen, from selfhood to nation, continues to challenge the legitimacy of the foretold metanarrative of national progression in which the countryside and inland cities belong to a bygone time that ought to be replaced in a forward-looking timeline. The explosive endings characteristic of Ying’s film oeuvre, in which all the imminent calamities happening to the region and its inhabitants eventually break out, belie the promise of a bright future told by the state and underline the symbolic violence endured by the local.

  3. Developing a Regional Recovery Framework

    Energy Technology Data Exchange (ETDEWEB)

    Lesperance, Ann M.; Olson, Jarrod; Stein, Steven L.; Clark, Rebecca; Kelly, Heather; Sheline, Jim; Tietje, Grant; Williamson, Mark; Woodcock, Jody

    2011-09-01

    Abstract A biological attack would present an unprecedented challenge for local, state, and federal agencies; the military; the private sector; and individuals on many fronts ranging from vaccination and treatment to prioritization of cleanup actions to waste disposal. To prepare the Seattle region to recover from a biological attack, the Seattle Urban Area Security Initiative (UASI) partners collaborated with military and federal agencies to develop a Regional Recovery Framework for a Biological Attack in the Seattle Urban Area. The goal was to reduce the time and resources required to recover and restore wide urban areas, military installations, and other critical infrastructure following a biological incident by providing a coordinated systems approach. Based on discussions in small workshops, tabletop exercises, and interviews with emergency response agency staff, the partners identified concepts of operation for various areas to address critical issues the region will face as recovery progresses. Key to this recovery is the recovery of the economy. Although the Framework is specific to a catastrophic, wide-area biological attack using anthrax, it was designed to be flexible and scalable so it could also serve as the recovery framework for an all-hazards approach. The Framework also served to coalesce policy questions that must be addressed for long-term recovery. These questions cover such areas as safety and health, security, financial management, waste management, legal issues, and economic development.

  4. Endoscopic anatomy of sellar region.

    Science.gov (United States)

    Perondi, Gerson Evandro; Isolan, Gustavo Rassier; de Aguiar, Paulo Henrique Pires; Stefani, Marco Antônio; Falcetta, E Frederico

    2013-06-01

    The transsphenoidal approach is the preferred access used in surgical treatment of most sellar region pathologies. The use of endoscopy is advantageous, and it is considered a good alternative to the traditional microsurgical technique. The purpose of this study is to recognize and describe anatomical variations of the sphenoid sinus and the sellar region, mainly describing the anatomy of the posterior wall of the sphenoid sinus and analyzing intercarotid distances in 3 regions. Thirty sphenoid blocks treated with formaldehyde were injected and dissected. Using endoscopy, anatomical variations were studied and the intercarotid distances were measured at the tuberculum sellae, sellar floor and clivus. The types of sphenoid sinus found were: conchal in 1 (4.76 %), pré-sellar in 2 (9.52 %) and sellar in 19 (85.7 %) specimens. The mean distance found from the sphenoid sinus ostium to the sella turcica was 19 mm (±6.5) mm. The mean intercarotid distances found at the tuberculum sellae, sellar floor and clivus were respectively 13.32, 18.00 and 18.90 mm. Endoscopy, with its magnification and lighting provide a panoramic view of deep fields. The anatomical variations described in this study support the need for a careful evaluation of preoperative images in each case.

  5. Regional projection of climate impact indices over the Mediterranean region

    Science.gov (United States)

    Casanueva, Ana; Frías, M.; Dolores; Herrera, Sixto; Bedia, Joaquín; San Martín, Daniel; Gutiérrez, José Manuel; Zaninovic, Ksenija

    2014-05-01

    Climate Impact Indices (CIIs) are being increasingly used in different socioeconomic sectors to transfer information about climate change impacts and risks to stakeholders. CIIs are typically based on different weather variables such as temperature, wind speed, precipitation or humidity and comprise, in a single index, the relevant meteorological information for the particular impact sector (in this study wildfires and tourism). This dependence on several climate variables poses important limitations to the application of statistical downscaling techniques, since physical consistency among variables is required in most cases to obtain reliable local projections. The present study assesses the suitability of the "direct" downscaling approach, in which the downscaling method is directly applied to the CII. In particular, for illustrative purposes, we consider two popular indices used in the wildfire and tourism sectors, the Fire Weather Index (FWI) and the Physiological Equivalent Temperature (PET), respectively. As an example, two case studies are analysed over two representative Mediterranean regions of interest for the EU CLIM-RUN project: continental Spain for the FWI and Croatia for the PET. Results obtained with this "direct" downscaling approach are similar to those found from the application of the statistical downscaling to the individual meteorological drivers prior to the index calculation ("component" downscaling) thus, a wider range of statistical downscaling methods could be used. As an illustration, future changes in both indices are projected by applying two direct statistical downscaling methods, analogs and linear regression, to the ECHAM5 model. Larger differences were found between the two direct statistical downscaling approaches than between the direct and the component approaches with a single downscaling method. While these examples focus on particular indices and Mediterranean regions of interest for CLIM-RUN stakeholders, the same study

  6. Regional air pollution over Malaysia

    Science.gov (United States)

    Krysztofiak, G.; Catoire, V.; Dorf, M.; Grossmann, K.; Hamer, P. D.; Marécal, V.; Reiter, A.; Schlager, H.; Eckhardt, S.; Jurkat, T.; Oram, D.; Quack, B.; Atlas, E.; Pfeilsticker, K.

    2012-12-01

    During the SHIVA (Stratospheric Ozone: Halogen Impacts in a Varying Atmosphere) campaign in Nov. and Dec. 2011 a number of polluted air masses were observed in the marine and terrestrial boundary layer (0 - 2 km) and in the free troposphere (2 - 12 km) over Borneo/Malaysia. The measurements include isoprene, CO, CO2, CH4, N2O, NO2, SO2 as primary pollutants, O3 and HCHO as secondary pollutants, and meteorological parameters. This set of trace gases can be used to fingerprint different sources of local and regional air pollution (e.g., biomass burning and fossil fuel burning, gas flaring on oil rigs, emission of ships and from urban areas, volcanic emissions, and biogenic emissions). Individual sources and location can be identified when the measurements are combined with a nested-grid regional scale chemical and meteorological model and lagrangian particle dispersion model (e.g., CCATT-BRAMS and FLEXPART). In the case of the former, emission inventories of the primary pollutants provide the basis for the trace gas simulations. In this region, the anthropogenic influence on air pollution seems to dominate over natural causes. For example, CO2 and CH4 often show strong correlations with CO, suggesting biomass burning or urban fossil fuel combustion dominates the combustion sources. The study of the CO/CO2 and CH4/CO ratios can help separate anthropogenic combustion from biomass burning pollution sources. In addition, these ratios can be used as a measure of combustion efficiency to help place the type of biomass burning particular to this region within the wider context of fire types found globally. On several occasions, CH4 enhancements are observed near the ocean surface, which are not directly correlated with CO enhancements thus indicating a non-combustion-related CH4 source. Positive correlations between SO2 and CO show the anthropogenic influence of oil rigs located in the South China Sea. Furthermore, SO2 enhancements are observed without any increase in CO

  7. The politics of inter-regionalism: Relations between international regional organizations

    NARCIS (Netherlands)

    Vleuten, J.M. van der; Ribeiro Hoffman, A.

    2013-01-01

    As the development of relations between international regional organizations, inter-regionalism denotes a relatively recent phenomenon. Largely due to systemic bipolarity, inter-regional relations remained limited to 'dialogue partnerships' between the European Community (EC) and other regional grou

  8. Regionalization procedures for hydrological drought assessment in Cevennes region, France

    Science.gov (United States)

    Dewaele, Helene; Vasiliades, Lampros; Esteves, Michel; Loukas, Athanasios

    2014-05-01

    Nowadays, low flow characteristics have been extensively studied for the design of hydrotechnical projects and water resources planning and management. Information on the magnitude and frequency of low flows is very important for hydrological drought analysis at operational level in public water supply systems. In this study, several low flow characteristics are derived and analysed for hydrological drought assessment at 26 watersheds located at Cevennes area, Southern France. The reliability of two statistical regionalisation techniques (the Principal Component Analysis and the L-moment approach) is tested on low flows characteristics for defining homogeneous regions in the study area and application of the methods to ungauged watersheds. The Threshold Level Method (TLM) is applied to estimate duration and deficit of the main hydrological drought events derived from flow duration curves at the study hydrometric stations using daily streamflow data for the period 1988 to 2008. Furthermore, the two regionalisation techniques are also applied in two other low flows indices, the annual minimum 7-days flow and the annual minimum 30-days flow. Finally, a regional analysis is performed in order to understand better the hydrological behavior of each watershed and the possible interactions between the hydrological regime and the descriptive hydrogeomorphologic characteristics.

  9. Hawaii Regional Sediment Management (RSM): Regional Sediment Budget for the West Maui Region

    Science.gov (United States)

    2016-06-01

    margin backed by steep mountainous terrain that has been vastly altered by agricultural and urbanized development. Shoreline hardening is being...majority of sediment transport (northward-directed in summer and southward-directed in winter). Because of these seasonal patterns, the net transport ...of sediment is small. There is evidence of nearshore eddy formation that increases the complexity of sediment transport in the region with sediment

  10. Regional energy integration in Africa

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2005-06-15

    This report is the first publication produced within the framework of the WEC's Africa Regional Action Plan as part of the 2005-2007 Work Programme. Presently, over 80% of the total energy consumption in Africa is based on traditional biomass used mostly for cooking. This lack of access to modern energy is holding back economic and social development for 1.6 billion people around the world. The situation is particularly grave in sub-Sahara Africa where over 80% of the population lives in rural areas and the average electrification rate is less than 5%. At least 50 million new connections are needed to provide electricity to supply the non-connected areas in Africa. The over 700 million potential customers represented by these new connections provide a major business opportunity. It is now widely recognised that development assistance, bilateral aid, multilateral financing institutions, a multitude of international aid agencies, NGOs and others have failed to make a significant difference. A new approach is required, otherwise the number of people without access to electricity will continue to grow, and none of the Millennium Development Goals set by the United Nations will be achieved. This regional report highlights key factors that affect cooperative energy projects. The geopolitical context, investment climate and appropriate regulation are just as important as the institutional and technical capacity required to execute many of these projects. The report identifies four key benefits of regional integration: improved security of supply and accessibility; increased economic efficiency; enhanced environmental quality and broader development of renewable resources.

  11. 47 CFR 90.1211 - Regional plan.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 5 2010-10-01 2010-10-01 false Regional plan. 90.1211 Section 90.1211... § 90.1211 Regional plan. (a) To facilitate the shared use of the 4.9 GHz band, each region may submit a plan on guidelines to be used for sharing the spectrum within the region. Any such plan must...

  12. Competitiveness of the region : content, factors, policies

    OpenAIRE

    2009-01-01

    The article describes a framework conception of regional competitiveness. The notion of regional competitiveness as well as approaches to the classification of competi-tiveness factors are considered in detail. The author elaborates a set of indicators of re-gional competitiveness. The article also dis-cusses three groups of factors which can influence the competitiveness of any region.

  13. Emergency Medicine in Remote Regions.

    Science.gov (United States)

    Renouf, Tia; Pollard, Megan

    2016-09-09

    Rural and remote places like Sable Island (Nova Scotia) or François (Newfoundland) pose a challenge in delivering both health care and appropriate education that today's learners need to practice in a rural setting. This education can be difficult to deliver to students far from academic centers. This is especially true for learners and practitioners at offshore locations like ships, oil installations, or in the air when patients are transported via fixed wing aircraft or helicopter. The following editorial provides a snapshot of the setting and the challenges faced while working as a physician on a ship, in remote regions.

  14. Tibet Benefiting From Regional Autonomy

    Institute of Scientific and Technical Information of China (English)

    GELEK

    2008-01-01

    Violent crimes involving beating, smashing,looting and burning have occurred in Lhasa,capital of Tibet Autonomous Region,and neighboring Qinghai,Gansu and Sichuan provinces in mid-March.Sufficient evidence shows that the riots were organized,premedi- tated,masterminded and incited by the Dalai Lama clique.For example,the Tibetan Youth Congress,a hardline organization under the Dalai Lama’s supporters that openly preaches violence,made a statement on March 10 this year saying that they would fight for"the independence of Tibet"defying death and sacrifices.

  15. Arms Race in Maghreb Region

    Institute of Scientific and Technical Information of China (English)

    EL AMOURI ALLAL

    2016-01-01

    Maghreb countries competitive altitude towards each other’ s has reached a higher level by entering an arms race.Morocco and Al ̄geria have dominated more than 50 percent of the Africa’ s imported weapons,mainly because of inherited cold war mentality of competi ̄tion and hostility. Maghreb countries competition has drugged the re ̄gion into a chaos that threatens regional stability obviously which af ̄fect the domestic political stability,since military spending weakens the financial capacity of states.

  16. Regional University Marketing in Under-developed Regions

    Directory of Open Access Journals (Sweden)

    Radovan Baèík

    2007-07-01

    Full Text Available Specific approaches, in the field of the University marketing, are discussed with the higher intensity last decades. But the Universities established in underdeveloped regions have special position, function, possibilities, and specific needed activities. There are also many reasons for applying modified strategies and marketing tools in Slovak higher educational market. The study deals with main and dominant approaches in this area of managing of educational institutions in Slovak environment but the study also presents the results of the research. The sample of the research was selected from students of the Faculty of Management, University of Preov, Slovak Republic. There was marketing product policy designed and mainly communications policy and image mainly analysed there. There are offered some suggestions for universities in the final part of the study.

  17. Surface chemistry in photodissociation regions

    CERN Document Server

    Esplugues, G B; Meijerink, R; Spaans, M; Caselli, P

    2016-01-01

    The presence of dust can strongly affect the chemical composition of the interstellar medium. We model the chemistry in photodissociation regions (PDRs) using both gas-phase and dust-phase chemical reactions. Our aim is to determine the chemical compositions of the interstellar medium (gas/dust/ice) in regions with distinct (molecular) gas densities that are exposed to radiation fields with different intensities. We have significantly improved the Meijerink PDR code by including 3050 new gas-phase chemical reactions and also by implementing surface chemistry. In particular, we have included 117 chemical reactions occurring on grain surfaces covering different processes, such as adsorption, thermal desorption, chemical desorption, two-body reactions, photo processes, and cosmic-ray processes on dust grains. We obtain abundances for different gas and solid species as a function of visual extinction, depending on the density and radiation field. We also analyse the rates of the formation of CO2 and H2O ices in d...

  18. Gastric Cancer Regional Detection System.

    Science.gov (United States)

    Ural, Berkan; Hardalaç, Fırat; Serhatlioğlu, Selami; İlhan, Mustafa Necmi

    2016-01-01

    In this study, a novel system was created to localize cancerous regions for stomach images which were taken with computed tomography(CT). The aim was to determine the coordinates of cancerous regions which spread in the stomach area in the color space with using this system. Also, to limit these areas with a high accuracy ratio and to feedback to the user of this system were the other objectives. This integration was performed with using energy mapping, analysis methods and multiple image processing methods and the system which was consisted from these advanced algorithms was appeared. For this work, in the range of 25-40 years and when gender discrimination was insignificant, 30 volunteer patients were chosen. During the formation of the system, to exalt the accuracy to the maximum level, 2 main stages were followed up. First, in the system, advanced image processing methods were processed between each other and obtained data were studied. Second, in the system, FFT and Log transformations were used respectively for the first two cases, then these transformations were used together for the third case. For totally three cases, energy distribution and DC energy intensity analysis were done and the performance of this system was investigated. Finally, with using the system's unique algorithms, a non-invasive method was achieved to detect the gastric cancer and when FFT and Log transformation were used together, the maximum success rate was obtained and this rate was calculated as 83,3119 %.

  19. Farmer Income Differential in Regions

    Institute of Scientific and Technical Information of China (English)

    LU Qian; MIAO Shanshan

    2006-01-01

    China's success in reform and opening up policy for twenty years is regarded as China's miracles in the world,whereas the income differential widening phenomenon has been the focus of the policymakers and researchers. This article researches 1994-2003 China's rural regions income differential and its decomposition. The method this paper used to measure the disparity is Gini Index. There are many ways to compute it, so the easiest way to decompose Gini Index-Matrix method is adopted. And based on it, farmer's income could be divided into wage income, farming income,transfer income and property income according to its composition. The conclusion is that all of the indexes are between 0.2 and 0.3, at the comparatively average level. From the fluctuation trend, it increased from 1994 to 1995, while reduced from 1995 to 1996, fluctuated in 1997, and then diminished again. In general, farmer's regions income differential stays at comparatively average level, but it has the widening trend with time. Through decomposing Gini Index, wage income is the most important increasing factor, while farming income is the reducing factor.

  20. Southern Appalachian Regional Seismic Network

    Energy Technology Data Exchange (ETDEWEB)

    Chiu, S.C.C.; Johnston, A.C.; Chiu, J.M. [Memphis State Univ., TN (United States). Center for Earthquake Research and Information

    1994-08-01

    The seismic activity in the southern Appalachian area was monitored by the Southern Appalachian Regional Seismic Network (SARSN) since late 1979 by the Center for Earthquake Research and Information (CERI) at Memphis State University. This network provides good spatial coverage for earthquake locations especially in east Tennessee. The level of activity concentrates more heavily in the Valley and Ridge province of eastern Tennessee, as opposed to the Blue Ridge or Inner Piedmont. The large majority of these events lie between New York - Alabama lineament and the Clingman/Ocoee lineament, magnetic anomalies produced by deep-seated basement structures. Therefore SARSN, even with its wide station spacing, has been able to define the essential first-order seismological characteristics of the Southern Appalachian seismic zone. The focal depths of the southeastern U.S. earthquakes concentrate between 8 and 16 km, occurring principally beneath the Appalachian overthrust. In cross-sectional views, the average seismicity is shallower to the east beneath the Blue Ridge and Piedmont provinces and deeper to the west beneath the Valley and Ridge and the North American craton. Results of recent focal mechanism studies by using the CERI digital earthquake catalog between October, 1986 and December, 1991, indicate that the basement of the Valley and Ridge province is under a horizontal, NE-SW compressive stress. Right-lateral strike-slip faulting on nearly north-south fault planes is preferred because it agrees with the trend of the regional magnetic anomaly pattern.

  1. Spicule Dynamics over Plage Region

    CERN Document Server

    Anan, Tetsu; Kawate, Tomoko; Matsumoto, Takuma; Ichimoto, Kiyoshi; Shibata, Kazunari; Hillier, Andrew; Otsuji, Kenichi; Watanabe, Hiroko; UeNo, Satoru; Nagata, Shin'ichi; Ishii, Takako T; Komori, Hiroyuki; Nishida, Keisuke; Nakamura, Tahei; Isobe, Hiroaki; Hagino, Masaoki

    2010-01-01

    We studied spicular jets over a plage area and derived their dynamic characteristics using Hinode Solar Optical Telescope (SOT) high-resolution images. The target plage region was near the west limb of the solar disk. This location permitted us to study the dynamics of spicular jets without the overlapping effect of spicular structures along the line of sight. In this work, to increase the ease with which we can identify spicules on the disk, we applied the image processing method `MadMax' developed by Koutchmy et al. (1989). It enhances fine, slender structures (like jets), over a diffuse background. We identified 169 spicules over the target plage. This sample permits us to derive statistically reliable results regarding spicular dynamics. The properties of plage spicules can be summarized as follows: (1) In a plage area, we clearly identified spicular jet features. (2) They were shorter in length than the quiet region limb spicules, and followed ballistic motion under constant deceleration. (3) The majorit...

  2. Regional case studies--Africa.

    Science.gov (United States)

    Prentice, Andrew M

    2009-01-01

    Africa is the final continent to be affected by the nutrition transition and, as elsewhere, is characterized by the paradoxical coexistence of malnutrition and obesity. Several features of the obesity epidemic in Africa mirror those in other emerging nations: it penetrates the richer nations and urban areas first with a strong urban- rural gradient; initially it affects the wealthy, but later there is a demographic switch as obesity becomes a condition more associated with poverty, and it shares many of the same drivers related to the increasing affordability of highly refined oils and carbohydrates, and a move away from subsistence farm work and towards sedentary lifestyles. Africa also has some characteristics of the obesity epidemic that stand out from other regions such as: (1) excepting some areas of the Pacific, Africa is probably the only region in which obesity (especially among women) is viewed culturally as a positive and desirable trait, leading to major gender differences in obesity rates in many countries; (2) most of Africa has very low rates of obesity in children, and to date African obesity is mostly an adult syndrome; (3) Africans seem genetically prone to higher rates of diabetes and hypertension in association with obesity than Caucasians, but seem to be relatively protected from dislipidemias; (4) the case-specific deaths and disabilities from diabetes and hypertension in Africa are very high due to the paucity of health services and the strain that the 'double burden' of disease places on health systems.

  3. Regional solid waste management study

    Energy Technology Data Exchange (ETDEWEB)

    1992-09-01

    In 1990, the Lower Savannah Council of Governments (LSCOG) began dialogue with the United States Department of Energy (DOE) regarding possibilities for cooperation and coordination of solid waste management practices among the local governments and the Savannah River Site. The Department of Energy eventually awarded a grant to the Lower Savannah Council of Governments for the development of a study, which was initiated on March 5, 1992. After careful analysis of the region`s solid waste needs, this study indicates a network approach to solid waste management to be the most viable. The network involves the following major components: (1) Rural Collection Centers, designed to provide convenience to rural citizens, while allowing some degree of participation in recycling; (2) Rural Drop-Off Centers, designed to give a greater level of education and recycling activity; (3) Inert landfills and composting centers, designed to reduce volumes going into municipal (Subtitle D) landfills and produce useable products from yard waste; (4) Transfer Stations, ultimate landfill disposal; (5) Materials Recovery Facilities, designed to separate recyclables into useable and sellable units, and (6) Subtitle D landfill for burial of all solid waste not treated through previous means.

  4. Evolution of active region outflows throughout an active region lifetime

    Science.gov (United States)

    Zangrilli, L.; Poletto, G.

    2016-10-01

    Context. We have shown previously that SOHO/UVCS data allow us to detect active region (AR) outflows at coronal altitudes higher than those reached by other instrumentation. These outflows are thought to be a component of the slow solar wind. Aims: Our purpose is to study the evolution of the outflows in the intermediate corona from AR 8100, from the time the AR first forms until it dissolves, after several transits at the solar limb. Methods: Data acquired by SOHO/UVCS at the time of the AR limb transits, at medium latitudes and at altitudes ranging from 1.5 to 2.3 R⊙, were used to infer the physical properties of the outflows through the AR evolution. To this end, we applied the Doppler dimming technique to UVCS spectra. These spectra include the H i Lyα line and the O vi doublet lines at 1031.9 and 1037.6 Å. Results: Plasma speeds and electron densities of the outflows were inferred over several rotations of the Sun. AR outflows are present in the newly born AR and persist throughout the entire AR life. Moreover, we found two types of outflows at different latitudes, both possibly originating in the same negative polarity area of the AR. We also analyzed the behavior of the Si xii 520 Å line along the UVCS slit in an attempt to reveal changes in the Si abundance when different regions are traversed. Although we found some evidence for a Si enrichment in the AR outflows, alternative interpretations are also plausible. Conclusions: Our results demonstrate that outflows from ARs are detectable in the intermediate corona throughout the whole AR lifetime. This confirms that outflows contribute to the slow wind.

  5. [Epidemiological imaginary in Campania Region].

    Science.gov (United States)

    Greco, Pietro

    2012-01-01

    The interviews on the epidemiological imaginary, collected within the framework of the project Sebiorec,(1) clearly demonstrate that also in Campania, on the border between the provinces of Naples and Caserta - where the issue of waste and land devastation take forms that are unprecedented compared to any other part of Europe - there is a widespread, strong, sacrosanct demand of participation in environment and health management. The request of deliberative ecological democracy is pressing.(2) There is an urgent need to meet that plethora of rights emerging in the "knowledge society" and in the "risk society" that someone has called "rights for scientific citizenship."(3) This request of the population of Campania, net of local cultural specificity, it is quite similar to that of the people of any other region of Europe. The context in which this request of participation is expressed, however, is quite different. Not only and not just for that real or perceived social pre-modern and familist web that would replace a modern civil society in Campania and all across the Southern Italian regions, but also and especially for some structural causes that we here try to list. Campania is a unique region in Europe - in many ways different even from other regions of southern Italy - due to the conjunction of at least five factors, not independent from each other. 1) The presence of a widespread organized crime which, in many areas, metropolitan and non-metropolitan alike, and especially in the provinces of Naples and Caserta, is a sort of state against the State and has one of its main levers of power and a major source of its wealth in the illegal control of the territory, in its different dimensions (military, but also economic, social and even cultural). 2) A huge social and economic disintegration, exacerbated in the last twenty years by a process of deindustrialization (until the early nineties Naples was the fifth industrial city of Italy, today it is a desert where

  6. Regional strategy tested in Caribbean.

    Science.gov (United States)

    1984-01-01

    Barbados, St. Vincent, and St. Lucia have joined forces in the world's 1st regional Contraceptive Social Marketing (CSM) effort -- the Caribbean CSM. The Barbados Family Planning Association (BFPS) is overseeing the operation, which begins selling 2 contraceptive pills and a condom in early February. Costs and start-up times were shaved by adopting brand names and advertising materials from Jamaica's highly successful CSM project. Jamaica's popular "Panther" condom and "Perle" oral contraceptive (OC) are being used by the Caribbean CSM project. Perle's 9-year-old package has been redesigned and the Caribbean CSM project also is selling a 2nd, low-dose version called "Perle-LD." The products are manufactured in the US by Syntex as Noriday and Norminest, respectively. But the regional approach's financial gains also had a debit side, most notably a tripling of bureaucratic procedures. Part of project difficulties stem from differences among the 3 Caribbean countries. While sharing a common cultural heritage, St. Lucians speak a patois dialect in addition to the English prevalent on the other islands. The biggest hurdle was overcoming an economic disparity between Barbados and its less affluent neighbors, St. Vincent and St. Lucia. The CSM project decided to try a 2-tier product pricing strategy. In US currency, prices run $1.75 per cycle for both OCs on Barbados, but $1.26 on St. Vincent and St. Lucia. A Panther 3-pack costs 75 cents on Barbados and 42 cents on the othe 2 islands. The project is being promoted with generic family planning media advertisements. The project also has held physician orientation seminars on each island. The pilot program will be accompanied by retailer training seminars. In addition the project may introduce a spermicidal foaming tablet, once the US Food and Drug Administration approvs a new American-made product. The unique Caribbean CSM project may spread an idea as potent as the family planning message. Its success could transmit the

  7. Cooperative monitoring of regional security agreements

    Energy Technology Data Exchange (ETDEWEB)

    Pregenzer, A.L.; Vannoni, M.; Biringer, K.L. [Sandia National Labs., Albuquerque, NM (United States). Nonproliferation and Arms Control Analysis Dept.

    1996-11-01

    This paper argues that cooperative monitoring plays a critical role in the implementation of regional security agreements and confidence building measures. A framework for developing cooperative monitoring options is proposed and several possibilities for relating bilateral and regional monitoring systems to international monitoring systems are discussed. Three bilateral or regional agreements are analyzed briefly to illustrate different possibilities. These examples illustrate that the relationship of regional or bilateral arms control or security agreements to international agreements depends on a number of factors: the overlap of provisions between regional and international agreements; the degree of interest in a regional agreement among the international community; efficiency in implementing the agreement; and numerous political considerations. Given the importance of regional security to the international community, regions should be encouraged to develop their own infrastructure for implementing regional arms control and other security agreements. A regional infrastructure need not preclude participation in an international regime. On the contrary, establishing regional institutions for arms control and nonproliferation could result in more proactive participation of regional parties in developing solutions for regional and international problems, thereby strengthening existing and future international regimes. Possible first steps for strengthening regional infrastructures are identified and potential technical requirements are discussed.

  8. Are temperature reconstructions regionally biased?

    CERN Document Server

    Bothe, O

    2012-01-01

    Are temperature reconstructions possibly biased due to regionally differing density of utilized proxy-networks? This question is assessed utilizing a simple process-based forward model of tree growth in the virtual reality of two simulations of the climate of the last millennium with different amplitude of solar forcing variations. The pseudo-tree ring series cluster in high latitudes of the northern hemisphere and east Asia. Only weak biases are found for the full network. However, for a strong solar forcing amplitude the high latitudes indicate a warmer first half of the last millennium while mid-latitudes and Asia were slightly colder than the extratropical hemispheric average. Reconstruction skill is weak or non-existent for two simple reconstruction schemes, and comparison of virtual reality target and reconstructions reveals strong deficiencies. The temporal resolution of the proxies has an influence on the reconstruction task and results are sensitive to the construction of the proxy-network. Existing ...

  9. Matching of equivalent field regions

    DEFF Research Database (Denmark)

    Appel-Hansen, Jørgen; Rengarajan, S.B.

    2005-01-01

    screen, having the same homogeneous medium on both sides and an impressed current on one aide, an alternative procedure is relevant. We make use of the fact that in the aperture the tangential component of the magnetic field due to the induced currents in the screen is zero. The use of such a procedure......In aperture problems, integral equations for equivalent currents are often found by enforcing matching of equivalent fields. The enforcement is made in the aperture surface region adjoining the two volumes on each side of the aperture. In the case of an aperture in a planar perfectly conducting...... shows that equivalent currents can be found by a consideration of only one of the two volumes into which the aperture plane divides the space. Furthermore, from a consideration of an automatic matching at the aperture, additional information about tangential as well as normal field components...

  10. C$_{60}$ in Photodissociation Regions

    CERN Document Server

    Castellanos, Pablo; Sheffer, Yaron; Wolfire, Mark G; Tielens, Alexander G G M

    2014-01-01

    Recent studies have confirmed the presence of buckminsterfullerene (C$_{60}$) in different interstellar and circumstellar environments. However, several aspects regarding C$_{60}$ in space are not well understood yet, such as the formation and excitation processes, and the connection between C$_{60}$ and other carbonaceous compounds in the interstellar medium, in particular polycyclic aromatic hydrocarbons (PAHs). In this paper we study several photodissociation regions (PDRs) where C$_{60}$ and PAHs are detected and the local physical conditions are reasonably well constrained, to provide observational insights into these questions. C$_{60}$ is found to emit in PDRs where the dust is cool ($T_d = 20-40$ K) and even in PDRs with cool stars. These results exclude the possibility for C$_{60}$ to be locked in grains at thermal equilibrium in these environments. We observe that PAH and C$_{60}$ emission are spatially uncorrelated and that C$_{60}$ is present in PDRs where the physical conditions (in terms of radi...

  11. [Chromosomal variation in Chironomus plumosus L. (Diptera, Chironomidae) from populations of Bryansk region, Saratov region (Russia), and Gomel region (Belarus)].

    Science.gov (United States)

    Belyanina, S I

    2015-02-01

    Cytogenetic analysis was performed on samples of Chironomus plumosus L. (Diptera, Chironomidae) taken from waterbodies of various types in Bryansk region (Russia) and Gomel region (Belarus). Karyotypes of specimens taken from stream pools of the Volga were used as reference samples. The populations of Bryansk and Gomel regions (except for a population of Lake Strativa in Starodubskii district, Bryansk region) exhibit broad structural variation, including somatic mosaicism for morphotypes of the salivary gland chromosome set, decondensation of telomeric sites, and the presence of small structural changes, as opposed to populations of Saratov region. As compared with Saratov and Bryansk regions, the Balbiani ring in the B-arm of chromosome I is repressed in populations of Gomel region. It is concluded that the chromosome set of Ch. plumosus in a range of waterbodies of Bryansk and Gomel regions is unstable.

  12. Evolution of active region outflows throughout an active region lifetime

    CERN Document Server

    Zangrilli, L

    2016-01-01

    We have shown previously that SOHO/UVCS data allow us to detect active region (AR) outflows at coronal altitudes higher than those reached by other instrumentation. These outflows are thought to be a component of the slow solar wind. Our purpose is to study the evolution of the outflows in the intermediate corona from AR 8100, from the time the AR first forms until it dissolves, after several transits at the solar limb. Data acquired by SOHO/UVCS at the time of the AR limb transits, at medium latitudes and at altitudes ranging from 1.5 to 2.3 R_sun, were used to infer the physical properties of the outflows through the AR evolution. To this end, we applied the Doppler dimming technique to UVCS spectra. These spectra include the H I Lyman alpha line and the O VI doublet lines at 1031.9 and 1037.6 A. Plasma speeds and electron densities of the outflows were inferred over several rotations of the Sun. AR outflows are present in the newly born AR and persist throughout the entire AR life. Moreover, we found two t...

  13. Generation Favorable Institutional Configuration Regional Business Environment

    Directory of Open Access Journals (Sweden)

    Natalia Zinovievna Solodilova

    2014-12-01

    Full Text Available This article discusses the theoretical issues of creating an enabling business environment, which is the base platform for the successful development of entrepreneurship in the regions. Provides A definition of a favorable institutional configuration of the regional business environment, which refers to forms of implementing the basic institutions and other regional institutions, taking into account existing regional system of formal and informal interaction between economic actors. States that despite the measures taken, the landscape of the Russian business community in terms of regions, remains uneven, with different indices of investment and business attractiveness, there is differentiation in business conditions in the regions with similar natural and geographical conditions and resource potential, which is primarily determined by , differences in the institutional configuration of the regional business environment and quality of interaction among the business community of the region. Hypothesis about the impossibility of creating a favorable business environment, institutional configurations at the same time in all regions of the country, as well as its limited duration. Conducted theoretical and probabilistic analysis of the parameters of creating an enabling institutional configuration of the business environment in the Russian regions. Grounded approach whereby institutional configuration of regional business environment, may be subject to management and control actions through targeted by the regional authorities can accept the specified (favorable to the business community parameters. The necessity of planning and effective management of a favorable institutional configuration of the business environment by regional authorities to increase the period of its existence.

  14. Regional characteristics of dust events in China

    Institute of Scientific and Technical Information of China (English)

    WANGShigong; WANGJinyan; ZHOUZijiang; SHANGKezheng; YANGDebao; ZHAO

    2003-01-01

    The regional characteristics of dust events in China has been mainly studied by using the data of dust storm,wind-blown sand and floating dust from 338 observation stations through China from 1954 to 2000.The results of this study are as follows:(1)In China,there are two high frequent areas of dust events,one is located in the area of Minfeng and Hotan in the South xinjiang Basin,the other is situated in the area of Minqin and Jilantai in the Hexi Region.Furthermore,the spatial distributions of the various types of dust events are different.The dust storms mainly occur in the arid and semiarid areas covering the deserts and the areas undergoing desertification in northern China.Wind-blown sand and floating-dust not only occur in the areas where dust storms occur,but also extend to the neighboring areas.The range of wind-blown sand extends northeastward and southeastward,but floating-dust mainly extends southeastward to the low-latitude region such as the East China Plain and the area of the middloe and lower reaches of the Yangtze River.Compared with wind-blown sand,the floating-dust seldom occurs in the high latitude areas such as North xinjiang and Northeast China.(2)The affected areas of dust storms can be divided into seven sub-regions,that is,North Xinjiang Region,South Xinjiang Region,Hexi Region,Qaidam Basin Region,Hetao Region.Northeastem China Region and Qinghai-Xizang (Tibet) Region.The area of the most frequent occurrence of dust storms and floating-dust is in South Xinjiang Region,and of wind-blown sang in the Hexi Region.In general,the frequency of dust events in all the seven regions shows a decreasing thendency from 1954 to 2000,but there are certain differences between various dust events in different regions.The maximum interannual change and ariance of dust events during this time happened in South Xinjiang Region and Hexi Region.The udst events generally occur most frequently in April in most parts of China.The spring occurred days of dust events

  15. Regional food culture and development.

    Science.gov (United States)

    Wahlqvist, Mark L; Lee, Meei-Shyuan

    2007-01-01

    Food culture is most influenced by the locality of its origin, which will have been one of food acquisition and processing by various means. It is generally agreed, and is the basis of much United Nations, especially Food and Agriculture Organisation strategic development policy, that successful agriculture, horticulture and aquaculture along with fishing, underpin economically viable and healthy communities with their various food cultures. We also know that this must be in tandem with maternal literacy and operational health care systems. These elements are best represented on a regional basis. There is a growing consumer interest in knowing where one's food comes from as a measure of "food integrity". However, food production alone can be a precarious business and relate to a lesser or greater extent to local food culture and to trade, which may be complementary or at-odds with each other. Likewise, the local food culture may have its strengths and weaknesses as far as its ability to meet nutritional and health needs is concerned. Local food production may be restricted because of geographical or socio-economic conditions which preclude food diversity, although this may be compensated for by trade. Where food adequacy and diversity is compromised, and soils poor, various macronutrient, micronutrient (from animals and plants) and phytonutrient (nutritionally-advantageous food component from plants) deficiencies may be in evidence. These food system problems may be intertwined with food culture--for example, "rice-based and water-soluble vitamin poor"; "few animal-derived foods like meat, fish, eggs and milk with associated low calcium, vitamin D, Vitamin B12 and long chain n-3 fatty acid intakes"; "low fruit and vegetable intake with limited carotenoids and other phytonutrients". Geo-satellite surveillance and mapping as identifying such "hot spots": for regional food problems, as well as hot spots where most of the world's biodiversity is found (1.4 % of land on

  16. What is Complex Regional Pain Syndrome?

    Science.gov (United States)

    ... on CRPS? Where can I get more information? What is complex regional pain syndrome? Complex regional pain ... almost no children under age 5 are affected. What are the symptoms of CRPS? The key symptom ...

  17. Enhancing regional security agreements through cooperative monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Pregenzer, A.L.

    1995-05-01

    This paper proposes that strengthening regional capabilities for formulating and implementing arms control and confidence-building measures is a tangible method of enhancing regional security. It discusses the importance of developing a regional infrastructure for arms control and confidence building and elucidates the role of technology in facilitating regional arms control and confidence-building agreements. In addition, it identifies numerous applications for regional cooperative monitoring in the areas of arms control, resource management, international commerce and disaster response. The Cooperative Monitoring Center at Sandia National Laboratories, whose aim is to help individual countries and regions acquire the tools they need to develop their own solutions to regional problems, is discussed briefly. The paper ends with recommendations for establishing regional cooperative monitoring centers.

  18. Region 7 Laboratory Information Management System

    Data.gov (United States)

    U.S. Environmental Protection Agency — This is metadata documentation for the Region 7 Laboratory Information Management System (R7LIMS) which maintains records for the Regional Laboratory. Any Laboratory...

  19. A SELF-ADAPTIVE TRUST REGION ALGORITHM

    Institute of Scientific and Technical Information of China (English)

    Long Hei

    2003-01-01

    In this paper we propose a self-adaptive trust region algorithm. The trust region radius is updated at a variable rate according to the ratio between the actual reduction and the predicted reduction of the objective function, rather than by simply enlarging or reducing the original trust region radius at a constant rate. We show that this new algorithm preserves the strong convergence property of traditional trust region methods. Numerical results are also presented.

  20. Helioseismic Ring Analysis of CME Source Regions

    Indian Academy of Sciences (India)

    S. C. Tripathy; S. de Wet; K. Jain; R. Clark; F. Hill

    2008-03-01

    We apply the ring diagram technique to source regions of halo coronal mass ejections (CMEs) to study changes in acoustic mode parameters before, during, and after the onset of CMEs.We find that CME regions associated with a low value of magnetic flux have line widths smaller than the quiet regions, implying a longer life-time for the oscillation modes. We suggest that this criterion may be used to forecast the active regions which may trigger CMEs.